EA039916B1 - Novel ferroportin inhibitors - Google Patents

Abstract

The invention concerns novel compounds having the general formula (A-II)pharmaceutical compositions comprising them and the use thereof as medicaments, in particular for the use as ferroportin inhibitors, more particularly for the use in the prophylaxis and/or treatment of diseases caused by a lack of hepcidin or iron metabolism disorders, such as particularly iron overload states ,such as in particular thalassemia and hemochromatosis.

Description

The field of technology to which the invention belongs

The present invention relates to new compounds having the general formula (A-I), preferably the general formula (A-II), pharmaceutical compositions containing these compounds, and their use as medicines, in particular use as inhibitors of ferroportin, more specifically - for use in the prevention and/or treatment of diseases caused by hepcidin deficiency or disorders of iron metabolism, such as, in particular, states of iron overload, such as, in particular, thalassemia and hemochromatosis.

Prior Art

Iron is an essential trace element for almost all organisms and is necessary, in particular, for the growth and formation of blood elements. The balance of iron metabolism in this case is regulated mainly at the level of return of iron from the hemoglobin of aging erythrocytes and absorption of iron from food in the duodenum. The released iron is absorbed in the intestine, in particular with the help of specific transport systems (DMT-1, ferroportin), enters the circulatory system and is delivered to the corresponding tissues and organs (transferrin, transferrin receptors).

In the human body, iron plays a huge role in, among other things, oxygen transport, oxygen uptake, cell functions such as electron transport in mitochondria, cognitive functions, etc., and energy metabolism in general.

On average, the human body contains 4-5 g of iron, which is present in enzymes, in hemoglobin and myoglobin, as well as in the form of deposited or reserve iron in the form of ferritin and hemosiderin. Approximately half of this iron, about 2 g, is heme iron bound in the hemoglobin of red blood cells. Since red blood cells have a limited lifespan (75-150 days), continuous production of new and removal of old red blood cells is necessary (over 2 million red blood cells are formed every second). A high regenerative capacity is provided by macrophages that absorb old erythrocytes through phagocytosis, lyse them and regenerate the iron thus obtained for further iron metabolism. This covers most of the daily iron requirement for erythropoiesis, which is 25 mg.

The daily iron requirement for an adult is 0.5 to 1.5 mg per day, children and pregnant women need 2-5 mg of iron per day. The daily loss of iron, for example due to the exfoliation of skin and epithelial cells, is small. Increased loss of iron is observed, for example, during menstrual bleeding in women. In general, blood loss can markedly lower iron levels, as approximately 1 mg of iron is lost per 2 milliliters of blood. In a healthy adult, the daily iron loss, which is normally about 1 mg, is usually replenished by a daily dietary intake, which replenishes the daily iron requirement to normal levels.

Iron levels are regulated by absorption, and the percentage of iron absorbed from food ranges from 6 to 12%; in the case of iron deficiency, the percentage of absorption increases to 25%. The level of absorption is regulated by the body depending on the need for iron and the size of iron stores. During this process, the human body absorbs both ferrous and ferric ions. Typically, iron(III) compounds dissolve in the stomach at sufficiently acidic pH values, and thus become available for absorption. Iron absorption is carried out in the upper part of the small intestine by mucosal cells. During this process, non-heme ferric iron is first reduced in the intestinal cell membrane to Fe(II) for absorption, for example by iron reductase (membrane-bound duodenal cytochrome b), so that it can then be transported into the cells intestine via the transport protein DMT1 (divalent metal transporter 1). On the contrary, heme iron enters enterocytes through the cell membrane without any changes. In enterocytes, iron is either stored in the composition of ferritin as deposited iron, or released into the blood by the transport protein ferroportin. Hepcidin plays a central role in this process as it is the most important regulator of iron absorption. The ferrous iron transported into the blood by ferroportin is converted into ferric iron by oxidases (ceruloplasmin, hephaestin), and then the ferric iron is transported to the corresponding parts of the body by transferrin (see, for example, Balancing acts: molecular control of mammalian iron metabolism. M.W. Hentze, Cell 117 , 2004, 285-297.)

Mammalian organisms are unable to actively excrete iron. Iron metabolism is mainly controlled by hepcidin through the cellular release of iron from macrophages, hepatocytes, and enterocytes.

Hepcidin is a peptide hormone produced in the liver. The predominant active form contains 25 amino acid residues (see, for example, Hepcidin, a key regulator of iron metabolism and mediator of anemia of inflammation. T. Ganz Blood 702,2003,783-8), although two forms truncated in amino end: hepcidin-22 and hepcidin-20. Hepcidin affects the absorption of iron through the intestines, through the placenta, and the release of iron from the reticuloendothelial system. In the body, hepcidin is synthesized from the so-called prohepcidin in the liver, and pro

- 1 039916 hepcidin is encoded by the so-called HAMP gene. The formation of hepcidin is regulated in direct correlation with the level of iron in the body, i.e. if there is enough iron and oxygen in the body, an increased amount of hepcidin is formed, and if the intake of iron and oxygen is low, or in case of increased erythropoiesis, less hepcidin is formed. In the mucosal cells of the small intestine and in macrophages, hepcidin binds to the transport protein ferroportin, through which phagocytally recycled iron is normally transported from the cells into the blood.

The transport protein ferroportin is a transmembrane protein containing 57 amino acid residues, which is formed in the liver, spleen, kidneys, heart, intestines and placenta. In particular, ferroportin is localized in the basolateral membrane of intestinal epithelial cells. Ferroportin bound in this way affects the export of iron into the blood. In this case, it is most likely that ferroportin transports iron in the form of Fe ²⁺ . If hepcidin is bound to ferroportin, ferroportin is transported into the cell, where it is destroyed, as a result of which the release of phagocytically recycled iron from the cells is then completely blocked. If ferroportin is inactivated, for example by hepcidin, so that it is not able to export the iron contained in the mucous cells, the iron contained in them is lost due to the natural excretion of the cells with the stool. Thus, intestinal absorption of iron is reduced when ferroportin is inactivated or suppressed, for example by hepcidin. In addition, ferroportin is markedly localized in the reticuloendothelial system (RES), which also includes macrophages. Hepcidin plays an important role in the deterioration of iron metabolism in chronic inflammation. In the case of such inflammations, in particular, the level of interleukin-6 increases, which leads to an increase in the content of hepcidin. As a result, an increased amount of hepcidin binds to macrophage ferroportin, thereby blocking the release of iron, which ultimately leads to the development of inflammation-induced anemia (ACD or AI).

On the other hand, if the iron content in the blood plasma decreases, then the production of hepcidin in the hepatocytes of the liver decreases, and therefore less hepcidin is released, correspondingly less ferroportin is inactivated, which allows more of the accumulated iron to be transported into the blood.

From the above, it becomes clear that the hepcidin-ferroportin system directly regulates iron metabolism and that a violation of the mechanism of hepcidin regulation directly affects iron metabolism in the body. In principle, the mechanism of hepcidin-ferroportin regulation operates on two opposite principles described below:

On the one hand, an increase in the content of hepcidin leads to the inactivation of ferroportin, which blocks the release of accumulated iron from cells into the blood, thereby lowering the iron content in the blood. In pathological cases, a reduced level of iron in the blood leads to a reduced content of hemoglobin, a decrease in the production of red blood cells and, as a result, to iron deficiency anemia.

On the other hand, a decrease in hepcidin leads to an increase in the content of active ferroportin, which allows to increase the release of stored iron and increase the intake of iron, for example from food, thereby increasing the level of iron in the blood. In pathological cases, an elevated level of iron in the blood leads to iron overload.

Iron overload conditions and diseases are characterized by excessive levels of iron. Problems arise due to excess iron in the blood, which leads to the appearance of the so-called free iron (NTBI). Free iron is rapidly nonspecifically absorbed by organs, which leads to the accumulation of iron in organs and tissues. Iron overload causes many diseases and unwanted medical conditions, including damage to the heart, liver, and endocrine system. In addition, iron accumulation in the brain has been observed in patients suffering from neurodegenerative diseases such as, for example, Alzheimer's disease and Parkinson's disease. As a particular detrimental aspect of excess free iron, undesirable formation of radicals should be mentioned. In particular, iron(III) ions catalyze the formation (among other things, by the Fenton reaction) of reactive oxygen species (ROS). ROS cause damage to DNA, fats, proteins, and carbohydrates, which has a long-term effect on cells, tissues, and organs. The formation of ROS is well known and described in the literature as the cause of the development of the so-called oxidative stress.

A well-known current treatment for iron overload is based on the concept of reducing the amount of iron in the blood by increasing the excretion of iron from the body. The oldest known and still routinely used method for otherwise healthy patients is regular phlebotomy (bleeding). After the first diagnosis, phlebotomy is usually prescribed quite frequently, for example once a week, until iron levels are in the normal range, after which phlebotomy is prescribed once a month or every three months, depending on the rate of accumulation of iron in a particular patient.

For patients unable to tolerate regular phlebotomy, chelating agents are available. For example, deferoxamine (also known as desferrioxamine B, N'-{5[acetyl(hydroxy)amino]pentyl}-N-[5-({4-[(5-aminopentyl)(hydroxy)amino]-4-oxobutanoyl} amino)

- 2 039916 pentyl]-N-hydroxysuccinamide or Desferal®), which is a bacterial siderophore known drug for use in chelation therapy. Deferoxamine binds iron in the circulatory system as a chelator and enhances its excretion in urine and feces. Typical treatment for chronic iron overload requires subcutaneous administration for 8-12 hours each day. Parenterally injectable compositions of desferrioxamine-B salts are described, for example, in WO 1998/25887.

Two new drugs licensed for use in patients who receive regular blood transfusions for the treatment of thalassemia that result in iron overload are deferasirox and deferiprone.

Deferasirox (Exjade®, 4-(3,5-bis(2-hydroxyphenyl)-1H-1,2,4-triazol-1-yl)benzoic acid) described for example in WO 1997/49395 and deferiprone ( Ferriprox®, 3-hydroxy-1,2-dimethylpyridin4(1H)-one) similarly function as iron chelating agents and are therefore suitable for use as drug compounds for iron chelation therapy.

Other compounds have also been described as iron chelators for the treatment of iron overload. For example, WO 2013/142258 concerns encapsulated particles of diethylenetriaminepentaacetate (DTPA) and a zinc salt. WO 2003/041709 relates to 4-hydroxy-2-alkylquinolines such as 4-hydroxy-2-nonylquinoline as iron chelators. WO 1998/09626 relates to chelating agents based on dithiocarbamate-containing compositions for the treatment of iron overload conditions.

WO 2015/077655 relates to desferritiocin derivatives having the general formula (A) or (J)

for use in the treatment of diseases associated with iron overload. According to WO 2015/077655, said desferritiocin derivatives have been found to work as iron chelating agents.

WO 2005/051411 relates to new antibiotics or antifungals based on oxachelin and its derivatives having the formula

which have been described as working as iron chelators and suitable for use in the treatment of diseases associated with iron overload.

The disadvantage of treating iron overload with chelation therapy is that the chelated iron is removed from the body when the iron overload has already occurred, instead of preventing the onset of the disease. In addition, the drugs used for iron chelation therapy are known to have toxic potential.

It can be expected that modern approaches will progressively replace this method, in particular with increasing knowledge of the underlying mechanisms and the development of suitable therapies based on this knowledge. Hepcidin agonists or compounds having an inhibitory or maintenance effect on the biochemical pathways regulating iron metabolism are generally known in the art.

Iron overload can occur, for example, when hepcidin production is hindered, for example due to a genetic defect, such as the well-known iron overload disease, hemochromatosis. Hemochromatosis is an iron overload disease caused by mutations in genes that control hepcidin synthesis or in the hepcidin gene itself. Low or no hepcidin levels in these patients result in elevated levels of active ferroportin, which enhances iron absorption from food, leading to severe iron overload causing damage to the heart, liver, and endocrine system. It was shown that hepcidin mimetic proteins, i.e. proteins that similarly bind and inactivate ferroportin effectively reverse tissue iron accumulation in gene-knockout mice

- 3 039916 hepcidin, Type 2 (juvenile) model of hemochromatosis. (Ramos et al., Blood 2012).

In a well-known iron overload disease, beta thalassemia, a mutation in the beta globin gene causes decreased hemoglobin production and ineffective erythropoiesis, the inability to produce enough red blood cells due to damage and death of the developing red blood cells in the bone marrow. This causes an increase in the rate of erythropoiesis and a decrease in hepcidin levels in order to make more iron available for the needs of increased erythropoiesis. This inadequate response results in iron overload due to reduced hepcidin levels, resulting in increased active ferroportin and enhanced dietary iron absorption as described above. Red blood cells in thalassemia have a shortened half-life due to the toxicity of an unbalanced ratio of hemoglobin alpha and beta subunits. Also, in the treatment of beta-thalassemia, the use of hepcidin mimetic proteins has been described, the therapeutic meaning of which is based on an increase in the activity of hepcidin, leading to a restriction of the level of iron and a decrease in iron-induced damage in red blood cells. Administration of hepcidin-mimetic proteins to th3/+ mice, in a transfusion-independent model of thalassemia, resulted in a reduction in the symptoms of ineffective erythropoiesis, an increase in the lifespan of red blood cells, and a reduction in anemia. In this model, prevention of iron overload due to reduced dietary iron absorption has been identified as an additional beneficial effect of hepcidin mimetic therapy (Gardenghi et al., 2010; Casu et al. 2013).

The described methods of therapy are based on direct intervention in the disturbed iron metabolism pathway through direct action through the primary regulator hepcidin through the administration of a hepcidin mimetic or hepcidin agonist, i.e. effects by replacing or increasing the amount of hepcidin. This approach is based on the therapeutic principle of treating iron overload, i. increased iron levels in the blood, by suppressing ferroportin through the mechanism of hepcidin inactivation, which leads to blocking of excess iron absorption.

Other known diseases associated with iron overload are those associated with ineffective erythropoiesis such as myelodysplastic syndrome (also known as MDS or myelodysplasia), polycythemia vera, etc.

In addition, mutations in genes involved in regulating systemic iron storage, such as hepcidin (Hamp1), hemochromatosis protein (HFE), hemojuvelin (HJV), and transferrin receptor 2 (TFR2), cause iron overload in mice and humans. Accordingly, diseases associated with mutations in the HFE gene, diseases associated with chronic hemolysis, sickle cell anemia, disorders of the erythrocyte membrane, as well as glucose-6phosphate dehydrogenase deficiency (G6PD deficiency), erythropoietic porphyria and Friedreich's ataxia can be indicated. In addition, subtypes of iron overload include transfusion iron overload, iron intoxication, pulmonary hemosiderosis, osteopenia, insulin resistance, African iron overload, Hallervorden-Spatz disease, hyperferritinemia, ceruloplasmin deficiency, neonatal hemochromatosis, and erythrocyte disorders including thalassemia, alpha thalassemia, middle thalassemia, sickle cell anemia, and myelodysplastic syndrome.

Other diseases and/or disorders and/or disease states associated with elevated iron levels include, but are not limited to, diseases caused by elevated iron levels, including ataxia, Friedreich's ataxia, age-related macular degeneration, age-related cataracts, age-related retinal diseases, and neurodegenerative diseases. diseases, where these neurodegenerative diseases include Alzheimer's disease, Parkinson's disease, pantothenatkinase-associated neurodegeneration, restless legs syndrome and Huntington's disease.

Hepcidin is a host defense protein that is a component of the innate immune system that responds to invading organisms.

Many bacteria have been reported to be highly dependent on iron supply from the host (so-called siderophilic organisms) and have developed mechanisms to capture iron from surrounding tissues. The ability to limit the amount of iron available to such organisms through the use of ferroportin inhibitors may be an effective adjuvant therapy. One such siderophilic organism is Vibrio vulnificus, which causes rare but extremely severe infections in coastal communities, often in patients with undiagnosed iron overload. Animal studies inoculated with lethal doses of Vibrio vulnificus have shown almost 100% survival when treated with ferroportin-inactivating hepcidin mimetic proteins, whether treated before or after infection (Arezes et al. 2015).

As known hepcidin mimetics, mention may be made of the so-called mini-hepcidins described, for example, in WO 2013/086143. Mini-hepcidins are small-sized synthetic peptide analogues of the N-terminus of hepcidin, which is responsible for the interaction of hepcidin with ferroportin. Mini-hepcidins were developed based on the discovery that the first 9 amino acids of hepcidin (DTHFPICIF) are sufficient to exhibit in vitro activity (measured by degradation of ferroportin-GFP). Mini-hepcidins contain a modified nine amino acid sequence

- 4 039916 hepcidin to increase resistance to proteolysis and enhance biophysical interaction with ferroportin. It has been described that mini-hepcidins can be used to treat iron overload conditions in humans caused by hepcidin deficiency.

WO 2015/069660 describes methods for enhancing hepcidin production for the treatment of iron overload conditions by reducing free iron (NTBI) by administering a modified iron-binding/releasing transferrin.

All of the described compounds that work as hepcidin agonists, hepcidin mimetics or ferroportin inhibitors, etc., are relatively high molecular weight compounds, especially those that are obtained mainly by genetic engineering. Various other approaches based on biomolecular interactions and biomolecules have been described. The disadvantages are the complex preparation and high sensitivity of such biomolecular compounds. In particular, methods based on antibodies to ferroportin are not effective enough, since ferroportin, suppressed by antibodies, is continuously reproduced by the body, and therefore the suppression is not long enough to achieve the desired therapeutic effect.

Also known are low molecular weight compounds that are involved in iron metabolism and can have an inhibitory or promoting effect.

For example, WO 2008/151288, WO 2008/118790, WO 2008/115999 and WO 2008/109840 describe compounds that act as divalent metal transporter 1 (DMT1) inhibitors and their use in the treatment of iron metabolism disorders such as thalassemia or hemochromatosis. .

WO 2008/123093 describes an agent for the prevention or treatment of iron overload conditions, which is 22 beta-methoxyolean-12-ene-3 beta,24(4 beta) diol.

EP 1074254 and EP 1072265 relate to the use of plant polyphenols with catechin and flavonoid structure for the treatment of iron overload.

WO 2011/029832 relates to thiazole and oxazole compounds which function as hepcidin antagonists and are therefore described as being suitable for the treatment of diseases associated with iron deficiency. This document describes that hepcidin antagonist activity inhibits the inhibition of ferroportin by hepcidin, which is the opposite effect found by the authors of the present invention for new thiazole and oxazole compounds.

Chemical compounds based on structures having the general formulas of the present invention have not been described to date for their activity as ferroportin inhibitors or for use in the prevention and treatment of iron metabolism disorders associated with elevated iron levels, such as iron overload.

In US 2004/0138268 A1, US 2011/0224136 A1, CN 103508957, WO 2006/062224 A1, WO 2015/051362 A1, EP 1953145 A1, WO 2009/154739 A2, GB 937807 A, WO 2017 2022 /020556 A1, WO 2005/011685 A1, WO 00/56724 A1, WO 2010/036632 A1, WO 2005/014576 A1, WO 2013/067578 A1, WO 2005/116355 A1 or Zou Yiquan et al. Discovery of pyrazole as C-terminus of selective BACE1 inhibitors; Eur. J. of Medicinal Chemistry 68 (2013) 270-283, Tussing-Humphreys et al. Rethinking Iron Regulation and Assessment in Iron Deficiency, Anemia of Chronic Disease, and Obesity: Introducing Hepcidin J. Academy of Nutrition and Dietetics (2012), Vol. 122, no. 3, 391-400, Riordan et al. Bleomycin analogs. Synthesis and proton NMR spectral assignments of thiazole amides related to bleomycin A2 (1); J. Heterocyclic Chem. 18, 1213 (1981), Hideaki Sasaki Synthesis of a novel bis(2,4'-bithiazole) derivative as a Co(II)activated DNA cleaving agent; Chem. Pharm. Bull. 42(8) 1685-1687 (1994), and Ballell et al. Fueling open-source drug discovery. 177 small-molecule leads against tuberculosis; ChemMedChem 2013, 8, 313-321 describes compounds for various medical applications and having different mechanisms of action.

The essence of the invention

The subject of the present invention is the development, in particular, of new therapeutically effective compounds that can be used for effective therapy for the prevention and treatment of disorders of iron metabolism associated with elevated iron levels, such as, in particular, iron overload. Within another subject of the invention, these new compounds should show few side effects and have very low toxicity and good bioavailability and compatibility. In addition, these new compounds, unlike known iron chelating compounds, should be suitable for preventing an increase in iron levels and thereby for the prevention of related diseases, and not for removing excess iron from the body when an increase in iron levels has already occurred. In another aspect of the invention, said novel compounds must have a defined structure (stoichiometry), be prepared using simple synthetic techniques, be less sensitive and have improved long-term efficacy compared to known biomolecular compounds such as antibodies.

Said subject matter has been achieved by developing new compounds having the formulas indicated herein, such as, in particular, formula (A-I), preferably formula (A-II), which have been found to work as inhibitors of ferroportin, and due to

- 5 039916 this can be used to suppress the transport of iron, and can be effectively used in the prevention and treatment of disorders of iron metabolism associated with elevated iron levels, such as, in particular, iron overload, as well as in the prevention and treatment of diseases caused by a lack of hepcidin, diseases associated with or caused by elevated iron levels or iron overload, and diseases associated with ineffective erythropoiesis.

Description of the invention

The present inventors have unexpectedly found that certain compounds having the general structural formula (A-I) work as inhibitors of ferroportin, thereby effectively inhibiting iron transport, and can be used as medicines, in particular for the treatment and/or prevention of diseases caused by deficiency hepcidin, diseases associated with ineffective erythropoiesis, or disorders of iron metabolism leading to an increase in iron levels, such as, in particular, states of iron overload, such as, in particular, thalassemia and hemochromatosis. In particular, the new compounds have proved to be suitable for the treatment of thalassemia and hemochromatosis. These new compounds can also be used to treat diseases caused by pathologically low levels of hepcidin and to suppress iron transport.

Accordingly, the present invention relates to novel compounds having the general formula (AI)

where Het-2 is an optionally substituted bicyclic heteroaryl having the general formula

where * indicates the point of attachment to A ² and

R4 means 1, 2 or 3 optional substituents which may be independently selected from the group consisting of the following representatives: a halogen atom, a cyano group, an optionally substituted alkyl group, an optionally substituted alkoxy group, and a carboxyl group;

X ¹ represents C, N, S or O;

X ² represents C or N;

X ³ represents C, N, S or O; and

X ⁴ is C, N or S, provided that 1 to 3 X heteroatoms are present, and where X ¹ , X ³ and X ⁴ , when they are C or N, may carry an additional substituent, such as, preferably, a hydrogen atom or a substituent as described above for a substituted heteroaryl;

R ¹ is selected from the group consisting of a hydrogen atom and optionally substituted alkyl; Cycl is selected from the group consisting of substituted aryl and substituted or unsubstituted heteroaryl; Q represents a hydrogen atom or C ₁ -C 4 -alkyl, which can form a conjugated 5 - or 6-membered cycle with Cycl; n is 0 or an integer from 1 to 8, preferably n is 0 an integer from 1 to 4, preferably n is 0, 1, 2 or 3;

A ¹ is an optionally substituted alkanediyl;

A ² is an optionally substituted alkanediyl or a direct bond;

R ³ represents a hydrogen atom or optionally substituted alkyl;

or A ¹ and R ³ together with the nitrogen atom to which they are attached form an optionally substituted 4-6 membered mono- or bicyclic ring;

or R ³ and A ² together with the nitrogen atom to which they are attached, form an optionally substituted 4-7-membered cycle; and

Y ² is C or N, where both Y ² can be C or one Y ² can be N and one Y ² can be C; or their pharmaceutically acceptable salts.

- 6 039916

In the description, the substituent groups mentioned above have the following meanings:

Optionally substituted alkyl preferably comprises linear or branched alkyl preferably containing 1-8, more preferably 1-6, particularly preferably 1-4, even more preferably 1, 2 or 3 carbon atoms.

Optionally substituted alkyl also includes cycloalkyl containing preferably 3-8, more preferably 5 or 6 carbon atoms.

Examples of alkyl residues having 1-8 carbon atoms include methyl group, ethyl group, n-propyl group, isopropyl group, n-butyl group, isobutyl group, sec-butyl group, tert-butyl group, n-pentyl group, isopentyl group, sec-pentyl group, tert-pentyl group, 2-methylbutyl group, n-hexyl group, 1-methylpentyl group, 2-methylpentyl group, 3-methylpentyl group, 4-methylpentyl group, 1-ethylbutyl group, 2-ethylbutyl group, 3-ethylbutyl group, 1,1dimethylbutyl group, 2,2-dimethylbutyl group, 3,3-dimethylbutyl group, 1-ethyl-1methylpropyl group, n-heptyl group, 1-methylhexyl group, 2-methylhexyl group, 3-methylhexyl group , 4-methylhexyl group, 5-methylhexyl group, 1-ethylpentyl group, 2-ethylpentyl group, 3-ethylpentyl group, 4-ethylpentyl group, 1,1-dimethylpentyl group, 2,2-dimethylpentyl group, 3,3-dimethylpentyl group pu, 4,4-dimethylpentyl group, 1-propylbutyl group, n-octyl group, 1-methylheptyl group, 2-methylheptyl group, 3-methylheptyl group, 4-methylheptyl group, 5-methylheptyl group, 6-methylheptyl group, 1-ethylhexyl group group, 2-ethylhexyl group, 3-ethylhexyl group, 4-ethylhexyl group, 5-ethylhexyl group, 1,1dimethylhexyl group, 2,2-dimethylhexyl group, 3,3-dimethylhexyl group, 4,4dimethylhexyl group, 5,5- dimethylhexyl group, 1-propylpentyl group, 2-propylpentyl group, etc. Groups containing 1-6, preferably 1-4 carbon atoms, such as in particular methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl and tert-butyl, are preferred. C ₁ -C ₃ alkyl, in particular methyl, ethyl and isopropyl, are more preferred. Most preferred are C1 and _C2 alkyl such as methyl and ethyl.

Cycloalkyl residues containing 3 to 8 carbon atoms preferably include a cyclopropyl group, a cyclobutyl group, a cyclopentyl group, a cyclohexyl group, a cycloheptyl group, and a cyclooctyl group. A cyclopropyl group, a cyclobutyl group, a cyclopentyl group and a cyclohexyl group are preferred. A cyclopentyl group and a cyclohexyl group are particularly preferred.

The substituents on the optionally substituted alkyl described above preferably comprise 1 to 3 identical or different substituents, more preferably 1 or 2 identical or different substituents selected, for example, from the group consisting of: optionally substituted cycloalkyl as defined above, hydroxy- group, oxo group (=O), carboxy group, halogen atom as defined below, cyano group, alkoxy group as defined below, optionally substituted acyl as defined below, optionally substituted acyloxy group, as defined below, optionally substituted aryl as defined below, optionally substituted heteroaryl as defined below, optionally substituted heterocyclyl as defined below, optionally substituted amino group as defined below, optionally substituted alkyl, aryl, or heterocyclyl a sulfonyl (R-SO2-) as defined below, as well as an alkylene group, such as in particular a methylene group, forming, for example, a methylene-substituted ethylene group (CH ₃ -(C=CH ₂ )- or where * indicates the point of connection). Preferably, 1-3 substituents on the alkyl are selected from the following: optionally substituted cycloalkyl, hydroxy group, oxo group (=O), carboxy group, optionally substituted acyloxy group, halogen atom, optionally substituted aryl, optionally substituted heteroaryl, optionally a substituted heterocyclyl, an optionally substituted amino group, an optionally substituted alkyl, aryl or heterocyclylsulfonyl (R-SO2-) and an alkylene group, such as in particular a methylene group. More preferred are 1-3 alkyl substituents selected from optionally substituted aryl, optionally substituted heteroaryl and optionally substituted heterocyclyl, and an alkylene group such as, in particular, a methylene group. More preferred is one substituent on the alkyl. Most preferred is one substituent on alkyl which is an optionally substituted aryl or an optionally substituted heteroaryl, as defined below.

In the context of the present invention, the halogen atom includes a fluorine, chlorine, bromine and iodine atom, preferably a fluorine or chlorine atom, most preferably a fluorine atom.

Examples of a linear or branched alkyl radical substituted with a halogen atom and containing 1 to 8 carbon atoms include a fluoromethyl group, a difluoromethyl group, a trifluoromethyl group, a chloromethyl group, a dichloromethyl group, a trichloromethyl group,

- 7 039916 bromomethyl group, dibromomethyl group, tribromomethyl group, 1-fluoroethyl group, 1-chloroethyl group, 1-bromoethyl group, 2-fluoroethyl group, 2-chloroethyl group, 2bromoethyl group, difluoroethyl group such as 1,2-difluoroethyl group, 1,2-dichloroethyl group, 1,2-dibromoethyl group, 2,2-difluoroethyl group, 2,2-dichloroethyl group, 2,2-dibromoethyl group, 2,2,2-trifluoroethyl group, heptafluoroethyl group, 1fluoropropyl group, 1-chloropropyl group, 1-bromopropyl group, 2-fluoropropyl group, 2-chloropropyl group, 2-bromopropyl group, 3-fluoropropyl group, 3-chloropropyl group, 3-bromopropyl group, 1,2-difluoropropyl group, 1,2-dichloropropyl group, 1,2-dibromopropyl group, 2,3-difluoropropyl group, 2,3-dichloropropyl group, 2,3-dibromopropyl group, 3,3,3-trifluoropropyl group, 2,2,3,3,3-pentafluoropropyl group, 2-fluorobutyl group , 2-chlorobutyl group, 2-bromobutyl group ppu, 4-fluorobutyl group, 4-chlorobutyl group, 4-bromobutyl group, 4,4,4trifluorobutyl group, 2,2,3,3,4,4,4-heptafluorobutyl group, perfluorobutyl group, 2fluoropentyl group, 2-chloropentyl group, 2-bromopentyl group, 5-fluoropentyl group, 5-chloropentyl group, 5-bromopentyl group, perfluoropentyl group, 2-fluorohexyl group, 2-chlorohexyl group, 2-bromohexyl group, 6-fluorohexyl group, 6-chlorohexyl group, 6- bromohexyl group, perfluoroheptyl group, 2-fluoroheptyl group, 2-chloroheptyl group, 2-bromoheptyl group, 7-fluoroheptyl group, 7chloroheptyl group, 7-bromoheptyl group, perfluoroheptyl group, etc. Particular mention should be made of fluoroalkyl, difluoroalkyl and trifluoroalkyl, with trifluoromethyl and mono- and difluoroethyl being preferred. Especially preferred are trifluoromethyl and 2,2-difluoroethyl.

Examples of the halogen-substituted cycloalkyl moiety containing 3 to 8 carbon atoms include 2-fluorocyclopentyl group, 2-chlorocyclopentyl group, 2-bromocyclopentyl group, 3-fluorocyclopentyl group, 3-chlorocyclopentyl group, 3-bromocyclopentyl group, 2-fluorocyclohexyl group, 2-chlorocyclohexyl group, 2-bromocyclohexyl group, 3-fluorocyclohexyl group, 3-chlorocyclohexyl group, 3-bromocyclohexyl group, 4-fluorocyclohexyl group, 4-chlorocyclohexyl group, 4-bromocyclohexyl group, difluorocyclopentyl group, dichlorocyclopentyl group, dibromocyclopentyl group, difluorocyclohexyl group, a dichlorocyclohexyl group, a dibromocyclohexyl group, a trifluorocyclohexyl group, a trichlorocyclohexyl group, a tribromocyclohexyl group, and so on.

Examples of the hydroxy-substituted alkyl moiety include the above alkyl moieties which contain 1-3 hydroxyl moieties, such as, for example, hydroxymethyl, 2-hydroxyethyl, 3-hydroxypropyl, etc. Hydroxymethyl is preferred.

Examples of the oxo-substituted alkyl moiety include the above alkyl moieties where at least one carbon atom is substituted by an oxo group forming a carbonyl group [-(C=O)-] in the alkyl chain or an alkanoyl group [alkyl-(C=O )-)], such as Ci-C ₆ alkanoyl such as formyl, acetyl, propionyl, butyryl, isobutyryl, valeryl, isovaleryl, pivaloyl, hexanoyl, etc. Preferably oxo substitution on the alkyl moiety in the form of a carbonyl group [-(C=O)-] or an acetyl group such as [-(C=O)-CH ₃ ] or [-(C=O)-CH ₂ -] .

Examples of the alkoxy-substituted alkyl moiety include the above alkyl moieties which contain 1-3 alkoxy moieties as defined below, such as, for example, methoxymethyl, ethoxymethyl, 2-methoxyethylene, etc.

Examples of the acyl-substituted alkyl moiety include the above alkyl moieties which contain 1-3 acyl moieties as defined below.

Examples of the acyloxy substituted alkyl moiety include the above alkyl moieties which contain 1-3, preferably 1 acyloxy moiety [-O-(C=O)-].

Examples of a cycloalkyl-substituted alkyl moiety include the above alkyl moieties containing 1-3, preferably 1 (optionally substituted) cycloalkyl group such as, for example, cyclopropylmethyl, cyclobutylmethyl, cyclopentylmethyl, cyclohexylmethyl, 2-cyclopropylethyl, 2-cyclobutylethyl, 2-cyclopentylethyl, 2-cyclohexylethyl, 2 - or 3-cyclopropylpropyl, 2- or 3-cyclobutylpropyl, 2- or 3-cyclopentylpropyl, 2- or 3-cyclohexylpropyl, etc. Preferred are cyclopropylmethyl and cyclohexylmethyl.

Examples of an aryl-substituted alkyl moiety include the above alkyl moieties containing 1-3, preferably 1 (optionally substituted) aryl group as defined below, such as, for example, phenylmethyl, 1- or 2-phenylethyl, 2- or 3- phenylpropyl, etc., phenylmethyl, 1-phenylethyl, 2-phenylethyl and 2-phenylpropyl are preferred. Also especially preferred are alkyl groups, as defined above, which are substituted by substituted aryl, as defined below, in particular by phenyl having 1-3, preferably 1 or 2 identical or different substituents, preferably selected from their halogen atom, such as preferably F and Cl, cyano, optionally substituted alkyl such as preferably methyl, ethyl, halo-substituted alkyl such as trifluoromethyl, optionally substituted alkoxy such as methoxy, ethoxy, halogen-substituted alkoxy

- 8 039916 a group such as a difluoromethoxy group, a trifluoromethoxy group, an optionally substituted amino group such as an amino group (NH2-) or a mono- or di-alkylamino group such as preferably a dimethylamino group, an optionally substituted heterocyclyl group such as pyrrolidinyl, alkyl-substituted piperazinyl or morpholinyl, or an optionally substituted heterocyclylsulfonyl group such as N-morpholinylsulfonyl, forming in particular alkyl groups which are substituted with substituted aryl according to the formulas

and which are particularly preferred for R ¹ and/or R2.

Examples of the heterocyclyl-substituted alkyl moiety include the above alkyl moieties containing 1-3, preferably 1 (optionally substituted) heterocyclyl group, as defined below, which may be substituted with 1-3, preferably 1 substituent. Preferably, the heterocyclyl group as a substituent on the alkyl is, for example, a morpholinyl group, a piperazinyl group, a piperidinyl group, etc. The heterocyclyl group as defined above may have a substituent, and the preferred substituent is an optionally substituted alkyl group, preferably a methyl or ethyl group, or a trifluoromethyl group. Especially preferred is a piperidinyl group and a methyl-substituted morpholinyl group.

Examples of the heteroaryl-substituted alkyl moiety include the above alkyl moieties containing 1-3, preferably 1 (optionally substituted) heteroaryl group as defined below, such as, for example, pyridinyl, pyridazinyl, pyrimidinyl, pyrazinyl, pyrazolyl, imidazolyl, benzimidazolyl, a thiophenyl or oxazolyl group such as pyridin-2-yl-methyl, pyridin-3-yl-methyl, pyridin-4-yl-methyl, 2-pyridin-2-yl-ethyl, 2-pyridin-1-yl-ethyl, 2 -pyridin-3-yl-ethyl, pyridazin-3-yl-methyl, pyrimidin-2-ylmethyl, pyrimidin-4-yl-methyl, pyrazin-2-yl-methyl, pyrazol-3-yl-methyl, pyrazol-4 -yl-methyl, pyrazol-5yl-methyl, imidazol-2-yl-methyl, imidazol-5-yl-methyl, benzimidazol-2-yl-methyl, thiophen-2-yl-methyl, thiophen-3-yl-methyl , 1,3-oxazol-2-yl-methyl.

Preferred is an alkyl group which is substituted with optionally substituted pyridazinyl, such as, in particular, pyridazin-3-yl-methyl and pyridazin-3-yl-ethyl, optionally substituted with pyridinyl, such as, in particular, optionally substituted pyridin-2-yl -methyl, pyridin-3-yl-methyl, pyridin-4-yl-methyl, 2-pyridin-2-yl-ethyl, 2-pyridin-1-yl-ethyl, 2-pyridin-3-yl-ethyl, especially preferably optionally substituted pyridin-2-yl-methyl and 2-pyridin-2-yl-ethyl, optionally substituted pyrazol-3-yl-methyl, pyrazol-4-yl-methyl, pyrazol-5-yl-methyl, pyrazol-3 -ylethyl, pyrazol-4-yl-ethyl, pyrazol-5-yl-ethyl. Particularly preferred is a substituted pyridinylalkyl such as substituted pyridinylmethyl or substituted pyridinylethyl, where 1, 2 or 3 substituents are selected from a halogen atom such as fluorine, C ₁ -C ₃ alkyl such as methyl, and trifluoromethyl . Especially preferred is a fluoro-substituted pyridinyl-alkyl, such as a fluoro-substituted pyridinyl-methyl or a fluoro-substituted pyridinyl-ethyl. Most preferred is the fluoro-substituted pyridinylmethyl having the formula

F

Examples of a heteroaryl-substituted alkyl group also include, in particular, a cycloalkyl radical as defined above which is linked to a heteroaryl substituent by conjugated ring formation with a heteroaryl substituent as defined above, preferably the conjugated cycloalkyl radical is cyclopentyl or cyclohexyl. Further, preferably, the conjugated heteroaryl substituent is pyridinyl, forming, for example, conjugated rings such as cyclopenta-pyridinyl and cyclohexa-pyridinyl having the formulas

which are particularly preferred for R ² , or a Cycl-[CQ]n group, where Q is a C1-C ₄ alkyl which forms a conjugated 5- or 6-membered cycle with Cycl.

In each case, the heterocyclyl substituent on the alkyl moiety as defined herein may be substituted with 1-3, preferably 1 or 2 identical or different substituents, which are preferably selected from a halogen atom such as preferably F and Cl, cyano

- 9 039916 group, optionally substituted alkyl such as preferably methyl, ethyl, halogenated alkyl such as trifluoromethyl and hydroxy-substituted alkyl such as hydroxymethyl, optionally substituted alkoxy group such as preferably methoxy and ethoxy group, oxo groups (=O), a heterocyclyl group as defined below, such as an N-morpholinyl group, an aminocarbonyl group, an optionally substituted amino group, such as preferably an amino group (NH2-), or a mono- or di-alkylamino group, such as, preferably a dimethylamino group.

Examples of the amino-substituted alkyl moiety include the above-described alkyl moieties containing 1-3, preferably 1 (optionally substituted) amino group as defined below, such as, for example, aminoalkyl (NH ₂ -alkyl) or mono- or dialkylamino- alkyl such as aminomethyl, 2-aminoethyl, 2- or 3-aminopropyl, methylaminomethyl, methylaminoethyl, methylaminopropyl, 2-ethylaminomethyl, 3-ethylaminomethyl, 2-ethylaminoethyl, 3-ethylaminoethyl, etc. or an alkyl group which may be substituted with an optionally substituted alkyloxycarbonylamino group such as a group having the formula o/—R where R is an alkyl substituent as defined above, preferably a phenyl group, such a group being particularly preferred for R ³ .

In the context of the present invention, optionally substituted aryl preferably includes: aromatic hydrocarbon residues containing 6-14 carbon atoms (excluding carbon atoms in possible substituents), which may be monocyclic or bicyclic, including, for example: phenyl, naphthyl, phenanthrenyl and anthracenyl, which may optionally be substituted, preferably with 1-3 identical or different substituents (for example, denoted R ⁶ ), selected from hydroxy, halogen atom, as defined above, cyano, optionally substituted amino, as defined below, optionally substituted an alkyl group as defined above, an optionally substituted acyl group as defined below, and an optionally substituted alkoxy group as defined below, an optionally substituted aryloxy group as defined below, an optionally substituted heterocyclyloxy group as defined below. and below, optionally substituted aryl, as defined herein, optionally substituted heterocyclyl, as defined below. Optionally substituted phenyl is preferred, such as unsubstituted phenyl and phenyl which is substituted with 1-3, more preferably 1 or 2 R ⁶ substituents, which may be the same or different. 1-3 phenyl substituents (for example, designated as R ⁶ ) are specifically selected from the group consisting of heterocyclyl, as defined below, halogen, as defined above, such as, in particular, F, optionally substituted amino group, definition which is given below, such as, in particular, (-NH ₂ ) or a mono- or dialkylamino group, with a dimethylamino group being preferred, a cyano group, an optionally substituted alkoxy group, the definition of which is given below, such as, in particular, a difluoromethoxy and trifluoromethoxy group, and an optionally substituted sulfonyl group which may in particular form a group wherein * denotes the binding site of a substituted phenyl substituent. Most preferred are halogen-substituted phenyl, alkoxy-substituted phenyl and hydroxyl-substituted phenyl. The above substituents on phenyl are particularly preferred for the Cycl group in the formulas herein, with the substituted aryl group being substituted phenyl.

Examples of an alkyl-substituted aryl group preferably include an aryl as defined above which is substituted by a linear or branched alkyl containing 1-8, preferably 1-4 carbon atoms, as described above. Toluoyl is the preferred alkylaryl.

Examples of a hydroxy-substituted aryl group preferably include an aryl as defined above which is substituted with 1-3 hydroxyl residues, such as, for example, 2hydroxyphenyl, 3-hydroxyphenyl, 4-hydroxyphenyl, 2,4-di-hydroxyphenyl, 2,5- dihydroxyphenyl, 2,6dihydroxyphenyl, 3,5dihydroxyphenyl, 3,6dihydroxyphenyl, 2,4,6trihydroxyphenyl, etc.

Examples of the halogen-substituted aryl group preferably include aryl as defined above which is substituted by 1-3 halogen atoms, such as, for example, 2-chloro- or fluorophenyl, 3-chloro- or fluorophenyl, 4-chloro- or fluorophenyl, 2 ,4-di-(chloro- and/or fluoro)phenyl, 2,5-di-(chloro- and/or

- 10 039916 fluoro)phenyl, 2,6-di-(chloro- and/or fluoro)phenyl, 3,5-di-(chloro- and/or fluoro)phenyl, 3,6-di-(chloro- and/ or fluoro)phenyl, 2,4,6-tri-(chloro- and/or fluoro)phenyl, etc.

Examples of the alkoxy-substituted aryl group preferably include aryl as defined above which is substituted with 1-3 alkoxy residues as described above, such as preferably 2-methoxyphenyl, 3-methoxyphenyl, 4-methoxyphenyl, 2-ethoxyphenyl, 3-ethoxyphenyl , 4ethoxyphenyl, 2,4-dimethoxyphenyl, etc., as well as difluoromethoxyphenyl and trifluoromethoxyphenyl.

In the context of the present invention, the optionally substituted heterocyclyl preferably comprises saturated or unsaturated mono- or bicyclic 4-8 membered heterocyclic residues which contain 1-3, preferably 1-2 identical or different heteroatoms selected from N, O and S and which may optionally be substituted, preferably with 1-3 substituents, the definitions of which are similar to substituents for optionally substituted heterocyclyl. 4-, 5-, and 6-membered saturated or unsaturated mono- or bicyclic optionally substituted heterocyclic residues are preferred, and examples thereof include azetidinyl, pyrrolidinyl, tetrahydrofuranyl, tetrahydrothiophenyl, piperidinyl, piperazinyl, tetrahydropyranyl, tetrahydrothiopyranyl, morpholinyl, etc., such as azetidin-1-yl, azetidin-2-yl, azetidin-3-yl, tetrahydrofuran-2-yl, tetrahydrofuran-3-yl, tetrahydro-thiophen-2-yl, tetrahydro-thiophen-3-yl, pyrrolidin-1- yl, pyrrolidin-2-yl, pyrrolidin-3-yl, morpholin-1-yl, morpholin-2-yl, morpholin-3-yl, piperidin-1-yl, piperidin-2-yl, piperidin-3-yl, piperidin-4-yl, piperazin-1-yl, piperazin-2-yl, tetrahydropyran-2yl, tetrahydropyran-3-yl, tetrahydropyran-4-yl, etc., which may optionally be fused with aromatic rings. Especially preferred are azetidinyl, pyrrolidinyl, piperidinyl and morpholinyl residues. Particularly preferred are the following heterocyclic residues, which may be substituted as described above:

(where X is N, O or S, preferably S), which are particularly preferred for A ¹ , and

especially preferred for R ¹ and/or R ² , and

which is particularly preferred as a substituent on the aryl group.

Preferred substituents on the heterocyclyl residues are an alkyl group such as preferably methyl and ethyl, a hydroxyl group and an oxo group (=O).

In the context of the present invention, optionally substituted heteroaryl includes heteroaromatic hydrocarbon residues containing 4-9 carbon atoms in the cycle, which further preferably contain 1-3 identical or different heteroatoms from the series S, O, N in the cycle, and therefore preferably form 5-12 -membered heteroaromatic residues, which may preferably be monocyclic, but also bicyclic. Preferred aromatic heterocyclic residues include pyridyl (pyridinyl), pyridyl X-oxide, pyridazinyl, pyrimidyl, pyrazinyl, thienyl (thiophenyl), furyl, pyrrolyl, pyrazolyl, imidazolyl, triazolyl, thiazolyl, oxazolyl or isoxazolyl, indolizinyl, indolyl, benzo[b ]thienyl, benzo[b]furyl, indazolyl, quinolyl, isoquinolyl, naphthyridinyl, quinazolinyl, quinoxalinyl. 5- or 6-membered aromatic heterocycles are preferred, such as members of the 5-membered heteroaryl group, for example thiazolyl such as thiazol-2-yl, 2-thiazol-2-yl, 2-thiazol-4-yl, thienyl (thiophenyl ) such as thien-3-yl, pyrazolyl such as 1-pyrazol-4-yl, 3-pyrazol-5-yl, imidazolyl such as imidazol-2-yl, 2-imidazol-4-yl, 1-imidazol- 4-yl, triazolyl such as 1-triazol-3-yl, 1-triazol-4-yl such as 1,2,4-triazol-3-yl or 1,2,3-triazol-4-yl, oxazolyl such as 2-oxazol-4-yl, 2-oxazol-5-yl, oxadiazolyl such as 1,2,4oxadiazol-3-yl, and members of the 6-membered heteroaryl group such as pyridyl (pyridinyl) such as pyride -1-yl, pyrid-2-yl, pyrid-3-yl, pyrid-4-yl, 2-pyrid-4-yl, 2-pyrid-6-yl, 3-pyrid-5-yl (pyridin-1 -yl, pyridin-2-yl, pyridin-3-yl, pyridin-4-yl, 2-pyridin-4-yl, 2-pyridin-6-yl, 3-pyridin-5- 11 039916 yl), pyrimidin- 2-yl, pyrimidin-4-yl, pyrimidin-5-yl, and members of the group of bicyclic heteroaromatic residues, in particular benzimidazolyl, such as benzimidazol-2-yl, benzimidazol-4yl, benzimidazol-5-yl, as well as benzimidazole-pyridinyl having the formula

or benzoxazol-2-yl having the formula

KP, or benzimidazole, forming a conjugated ring with a heterocyclyl residue, as defined above.

The above heteroaryl groups may have one or more, preferably 1-3, more preferably 1 or 2 identical or different substituents, which are in particular selected from a halogen atom, such as preferably F and Cl, cyano, optionally substituted alkyl, as defined above, such as preferably methyl, ethyl, n-propyl, isopropyl, halogen-substituted alkyl such as difluoromethyl or trifluoromethyl, hydroxy-substituted alkyl such as hydroxymethyl, aminocarbonyl-substituted alkyl such as aminocarbonylmethyl, carboxyl -substituted alkyl such as carboxylmethyl, alkenyl group such as propenyl, optionally substituted alkoxy group such as preferably methoxy and ethoxy group, hydroxyl group (-OH), oxo group (=O), carboxyl group [-(C=O)-OH], a heterocyclyl group as defined above such as an N-morpholinyl group, an aminocarbonyl group such as NH ₂ -(C=O)- is optionally substituted an amino group such as preferably an amino group (NH2-) or a mono- or di-alkylamino group such as preferably a dimethylamino group.

In particular, examples of an alkyl-substituted heteroaryl group preferably include the above-described heteroaryl which is substituted by a linear or branched, optionally substituted alkyl-described above containing 1-8, preferably 1-4 carbon atoms, such as, in particular, methylimidazolyl, such as in particular N-methylimidazolyl, methylbenzimidazolyl, such as in particular N-methylbenzimidazolyl, 5-methylbenzimidazolyl, 4-trifluoromethylbenzimidazolyl, 5trifluoromethylbenzimidazolyl,

N-aminocarbonylmethylbenzimidazolyl, N-carboxymethylaminocarbonyl, N-methylpyrazolyl, 1(N),5-dimethylpyrazolyl, methylpyridinyl such as 2-methylpyridin-3-yl, 2-methylpyridin-4-yl, 3methylpyridin-2-yl, 3-methylpyridine -3-yl, 3-methylpyridin-4-yl, 4-methylpyridin-2-yl, 5methylpyridin-2-yl, 6-methylpyridin-2-yl, etc., dimethylpyridinyl such as 3,5-dimethylpyridin- 2yl, 4,6-dimethylpyridin-3-yl, trifluoromethylpyridinyl, in particular 3- or 4-trifluoromethylpyridin-2yl, 6-trifluoromethylpyridin-3-yl, 3-hydroxymethylpyridin-2-yl, 5-methylpyrimidin-2-yl, etc. .d.

Examples of the halogen-substituted heteroaryl group preferably include the heteroaryl described above which is substituted by 1-3, preferably 1 or 2 halogen atoms, such as preferably F and/or Cl, including in particular a fluoropyridinyl such as 3-fluoro-pyridine -2-yl, 4-fluoropyridin-2-yl, 5-fluoro-pyridin-2-yl, 6-fluoro-pyridin-2-yl, 3-chloro-pyridin-2-yl, 4-chloro-pyridin-2 -yl, 5-chloro-pyridin-2-yl, 6-chloro-pyridin-2-yl, 2-fluoro-pyridin-3-yl, 4-fluoro-pyridin-3-yl, 5-fluoropyridin-3-yl , 6-fluoro-pyridin-3-yl, 2-chloro-pyridin-3-yl, 4-chloro-pyridin-3-yl, 5-chloro-pyridin-3-yl, 6-chloro-pyridin-3-yl , 2-fluoro-pyridin-4-yl, 3-fluoro-pyridin-4-yl, 5-fluoro-pyridin-4-yl, 6-fluoro-pyridin-4-yl, 2-chloro-pyridin-4-yl, 3 -chloro-pyridin-4-yl, 5-chloro-pyridin-4-yl, 6-chloro-pyridin-4-yl, etc., difluoropyridinyl such as 3,5-difluoropyridin-2-yl, fluoro- chloro-pyridinyl such as 3-chloro-5fluoro-pyridin-2-yl, etc.

Examples of the halogen- and alkyl-substituted heteroaryl group preferably include the above-described heteroaryl which is substituted by 1-3 halogen atoms, such as preferably F and/or Cl, and 1-3 linear or branched, optionally substituted by the alkyl radicals described above, such such as, in particular, 3-fluoro-6-methylpyridin-2-yl, 3-chloro-5-trifluoromethylpyridin2-yl.

More preferred examples of substituted heteroaryl groups include methoxypyridinyl such as 3-, 4-, 5- or 6-methoxypyridin-2-yl, 2-, 4-, 5- or 6-methoxypyridin-3-yl, 2-, 3- , 5- or 6-methoxypyridin-4-yl etc., hydroxypyridinyl such as 3-, 4-, 5- or 6-hydroxypyridin-2-yl, 2-, 4-, 5- or 6-hydroxypyridin-3 -yl, 2-, 3-, 5- or 6-hydroxypyridin-4-yl, etc., oxo-pyridinyl such as 6-oxo-1,6-dihydropyridin-2-yl, 2-oxo-1 ,2-dihydropyridin-3-yl, etc., aminopyridinyl such as 6dimethylaminopyridin-3-yl, aminocarbonylpyridinyl such as 6-aminocarbonylpyridin-3-yl, cyanopyridinyl such as 3-, 4-, 5- or 6 -cyanopyridin-2-yl, 2-, 4-, 5- or 6-cyanopyridin-3-yl, 2-, 3-, 5 or 6-cyanopyridin-4-yl, etc., as well as 2-morpholin- 4-yl-pyridin-4-yl.

When R ⁴ represents 1-3, preferably 1 or 2 identical or different substituents on the Het-2 bicyclic heteroaryl group according to any of the general formulas herein, said heteroaryl substituents are preferably selected from a halogen atom such as preferably F and Cl, cyano, optionally substituted alkyl as defined above, such as preferably methyl, ethyl, n-propyl, isopropyl, halogenated alkyl such as difluoromethyl or trifluoromethyl, aminocarbonyl-substituted alkyl such as aminocarbonylmethyl, a carboxyl-substituted alkyl such as carboxylmethyl, an optionally substituted alkoxy group such as preferably a methoxy and ethoxy group, and a carboxyl group [-(C=O)-OH]. Most preferably, R ⁴ is 1 or 2 identical or different substituents selected from F, Cl, cyano, optionally substituted alkyl such as methyl and trifluoromethyl, aminocarbonyl-substituted alkyl such as aminocarbonylmethyl, carboxyl-substituted alkyl such as carboxylmethyl, an optionally substituted alkoxy group such as a methoxy group; and a carboxyl group [-(C=O)-OH].

When R ⁵ is 1-4, preferably 1-3, more preferably 1 or 2 identical or different substituents on the heteroaryl group Cycl according to any of the general formulas herein, said heteroaryl substituents are preferably selected from a halogen atom such as preferably F and Cl, cyano, optionally substituted alkyl as defined above, such as preferably methyl, ethyl, n-propyl, isopropyl, halogen-substituted alkyl such as difluoromethyl or trifluoromethyl, hydroxy-substituted alkyl such as hydroxymethyl , an optionally substituted alkoxy group such as preferably a methoxy and ethoxy group, an oxo group (=O), a heterocyclyl group as defined above such as an N-morpholinyl group, an aminocarbonyl group such as NH ₂ -(C =O)-, an optionally substituted amino group such as preferably an amino group (NH2-) or a mono- or dialkylamino group such as preferred flax dimethylamino group. Most preferably R ⁵ is 1 or 2 identical or different substituents selected from F, Cl, cyano, optionally substituted alkyl such as methyl, trifluoromethyl and hydroxymethyl, optionally substituted alkoxy such as methoxy, oxo (=O) group forming, for example, an oxo-substituted heteroaryl having the general formula

a heterocyclyl group such as an N-morpholinyl group, an aminocarbonyl group such as H2-(C=O)-, an optionally substituted amino group such as a di-alkylamino group such as a dimethylamino group.

Optionally substituted acyl herein and hereinafter includes formyl (-CH(=O)), optionally substituted aliphatic acyl (alkanoyl=alkyl-CO, where optionally substituted alkyl is as defined above for the alkyl group), optionally substituted aromatic acyl ( aroyl = aryl-CO-, where optionally substituted aryl is as defined above for the aryl group), optionally substituted heteroaromatic acyl (heteroaroyl = heteroaryl-CO-, where optionally substituted heteroaryl is as defined above for the heteroaryl group), or heterocyclic acyl (heterocycloyl = heterocyclyl-CO-, where optionally substituted heterocyclyl is as defined above for the heterocyclyl group). Aliphatic acyl=alkanoyl=alkyl-CO- is preferred.

The optionally substituted amino group of the present invention preferably includes: an amino group (-NH2), an optionally substituted mono- or dialkylamino group (alkyl-NH-, (alkyl) ₂ N-), where alkyl has the above definition for optionally substituted alkyl. Also included are optionally substituted mono- or diarylamino, mono- or diheteroarylamino and mono- or diheterocyclylamino radicals or mixed optionally substituted alkylarylamino, alkylheteroarylamino and alkylheterocyclylamino radicals, where the above definitions for optionally substituted alkyl, aryl, heteroaryl and heterocyclyl apply. According to the present invention, the amino group also includes the -NH- group.

The optionally substituted amino group is preferably an optionally substituted mono- or dialkylamino group (alkyl-NH-, (alkyl) ₂ N-), in particular with 1-8, preferably 1-6, more preferably 1-3 carbon atoms, as mentioned above. The most preferred optionally substituted amino group is a mono- or dimethylamino group and a mono- or ethylamino group. Most preferred is an amino group (-NH ₂ ) or (-NH-) and a dimethylamino group.

In the context of the present invention, the optionally substituted alkanediyl is preferably a divalent linear or branched alkanediyl radical containing 1-7, preferably 1-6, more preferably 1-4 carbon atoms, which may optionally have 1-3, preferably 1 or 2 substituents, selected from the group consisting of a halogen atom, a hydroxy group, an oxo group (forming a carbonyl or acyl group) and an amino group,

- 13 039916 as defined above. Preferred examples include: methylene, ethane-1,2-diyl, ethane-1,1-diyl, propane-1,3-diyl, propane-1,1-diyl, propane-1,2-diyl, propane -2,2-diyl, butane-1,4-diyl, butane-1,2-diyl, butane-1,3-diyl, butane-2,3-diyl, butane-1,1-diyl, butane-2 ,2-diyl, butane-3,3diyl, pentane-1,5-diyl, etc. Particular preference is given to methylene, ethane-1,2-diyl, ethane-1,1-diyl, propane-1,3-diyl, propane-2,2-diyl and butane-2,2-diyl. Most preferred are methylene and ethane-1,2-diyl.

The preferred substituted alkanediyl radical is a hydroxy-substituted alkanediyl such as a hydroxyl-substituted ethanediyl, an oxo-substituted alkanediyl such as an oxo-substituted methylene or ethanediyl radical forming a carbonyl or acyl (acetyl) group, a halogen-substituted alkanediyl group such as an alkanediyl group, substituted with one or two halogen atoms selected from F and Cl, preferably a 2,2-difluoroethanediyl, or alkanediyl group which is substituted with an oxo and amino group, forming an aminocarbonyl group such as preferably a [-(C=O)-NH -].

According to the present invention, the substituents R ¹ and R ² or the corresponding group -[CQ]n-, where Q represents C1-C4 alkyl, may, together with the nitrogen atom to which they are attached, form an optionally substituted 3-6-membered ring , which may optionally contain additional heteroatoms. R ¹ and R ² (or the group -[CQ]n-, where Q is C1-C ₄ alkyl) may preferably, together with the nitrogen atom to which they are attached, form a 5- or 6-membered ring, which may contain additional heteroatoms, preferably one additional heteroatom selected from N and O. Most preferably, R ¹ and R ² (or the group -[CQ] _n -, where Q represents C 1 -C ₄ -alkyl) together with the nitrogen atom to which they are attached, forms a 6-membered ring that contains no additional heteroatoms, forming an N-piperidinyl ring, or a 6-membered ring that contains one additional O heteroatom, forming an N-morpholinyl ring. In particular, such N-piperidinyl ring may be substituted with aryl or heteroaryl as defined above, preferably with phenyl or piperidinyl, forming a bicyclic structure according to the formulas

or

According to the present invention, it is also possible that A ¹ when it is a linear or branched alkanediyl group as defined above and R ³ when it is an optionally substituted alkyl group as defined above, together with the nitrogen atom, to to which they are attached form an optionally substituted 4-6 membered aliphatic mono- or bicyclic ring which may be substituted with 1-3 substituents as defined above, such as, for example, substituents according to the following

telny.

In the context of the present invention, it is also possible that R ³ and A ² together with the nitrogen atom to which they are attached form an optionally substituted 4-7 membered ring, where the optional substituents are preferably selected from heteroaryl as defined above and an oxo group. The heteroaryl substituent may also form a fused ring with a 4-7 membered ring formed by R ³ and A ² together with the nitrogen atom to which they are attached. Examples include residues corresponding to the formulas below:

- 14 039916

concerns new compounds having the general formula

In another aspect, the present invention (A-II)

cycle

R where R ⁴ means 0, 1, 2 or 3 substituents independently selected from the group consisting of the following representatives:

halogen atom, cyano group, linear or branched C ₁ -C ₃ -alkyl bearing 0, 1, 2 or 3 halogen atoms,

C ₁ -C ₂ -alkoxy group and carboxyl group;

X ¹ , X ² , X ³ and X ⁴ are selected from C, N, S or O to form a group selected from:

and where X ¹ , X ³ and X ⁴ when they represent C or N, are not substituted or carry an additional substituent selected from halogen, C ₁ -C ₃ -alkyl bearing 0, 1, 2 or 3 halogen atoms; and where

Y ¹ represents a hydrogen atom, halogen or

C ₁ -C ₃ -alkyl, which carries 0, 1, 2 or 3 halogen atoms;

R ¹ is selected from the group consisting of a hydrogen atom and linear or branched C ₁ -C ₃ -alkyl;

Cycl is selected from the group consisting of substituted or unsubstituted 5- or 6-membered monocyclic heteroaryl containing 1-3 heteroatoms selected from S, O and N, where possible substituents are selected from halogen, cyano, unsubstituted C ₁ -C ₃ - αkyl or C ₁ -C ₃ -alkyl, which carries 0, 1, 2 or 3 halogen atoms or hydroxy groups, C ₁ -C ₃ -alkoxy, oxo groups (=O), forming an oxo-substituted heteroaryl of the formula

, N-morpholinyl groups, NH ₂ -(C=O)- aminocarbonyl groups, or dimethylamino groups;

Q is a hydrogen atom or

C1- _C4 alkyl which can form a conjugated 5- or 6-membered cycle with Cycl;

n is 0 or an integer from 1 to 2;

- 15 039916

A ¹ is an unsubstituted linear C1-C ₄ -alkanediyl;

A ² is a linear or branched unsubstituted or substituted C1-C4alkanediyl, where possible substituents are selected from 1 or 2 halogens, and oxo groups or a direct bond;

R ³ represents a hydrogen atom or an unsubstituted or substituted linear C ₁ -C ₃ -alkyl, where possible substituents are selected from the benzimidazolyl group, the cyclopropyl group and the amino NH ₂ group; or

And ¹ and R ³ together with the nitrogen atom to which they are attached, form an unsubstituted or substituted 4-6-membered mono - or bicyclic ring, where possible substituents are selected from methyl, ethyl and hydroxyl group; or

R ³ and A ² together with the nitrogen atom to which they are attached form an unsubstituted or substituted 4-membered ring, where the possible substituents are selected from a heteroaryl containing from 1 to 3 heteroatoms selected from S, O and N, and an oxo group ;

or its pharmaceutically acceptable salts.

Preferred Embodiments

Embodiment A-2.

A more preferred embodiment of the present invention relates to novel compounds having the formula (A-II)

have the meanings given above for the formula (A where Cycl, Q, R ¹ , X ¹ , X ² , X ³ , X ⁴ , R ³ , A ¹ , A ² , R ⁴ and n I); or their pharmaceutically acceptable salts.

Embodiment A-3.

A more preferred embodiment of the present invention relates to novel compounds having formula (AI) and (A-II) as described above, wherein Cycl is a substituted or unsubstituted heteroaryl as defined above. Heteroaryl may be substituted with 1-4, preferably 1-3, more preferably 1 or 2 R ⁵ substituents as defined above and below in the context of compounds having the general formulas (A-IIIa) (A-IIIb), (A-IVa ), (A-IVb), (A-IVc), and (A-IVd).

Embodiment A-3b.

A more preferred embodiment of the present invention relates to novel compounds having formula (A-I) and (A-II) as described above, wherein Cycl is a substituted or unsubstituted heteroaryl which is selected from substituted or unsubstituted pyridinyl, to form compounds having the formula (A-IIIb)

where Q, R ¹ , X ¹ , X ² , X ³ , X ⁴ , R ³ , A ¹ , A ² , Y ² , R ⁴ and n are as defined above for formula (A-I) or (A- II), and where R ⁵ means 1-4, preferably 1-3, more preferably 1 or 2 optional substituents, which may be independently selected from the group consisting of the following representatives:

halogen atom, preferably F and Cl, optionally substituted alkyl, preferably methyl, trifluoromethyl, hydroxymethyl, hydroxy group, alkoxy group, preferably methoxy group, oxo group (=O) forming a substituted pyridinyl group having the general formula

an amino group such as -NH2, a mono- or dialkylamino group, preferably a dialkylamino group, an aminocarbonyl group, preferably NH ₂ -(C=O)-, a cyano group, and

- 16 039916 a heterocyclyl group, preferably a morpholinyl group, or their pharmaceutically acceptable salts.

Embodiment A-4b.

A more preferred embodiment of the present invention relates to novel compounds having the formula (A-IVb) o

(A-IVb) where Q, R ¹ , X ¹ , X ² , X ³ , X ⁴ , R ³ , A ¹ , A ² , Y ² , R ⁴ and n are as defined above for formula (A-I ), (A-II) or (A-IIIb), and where R ⁵ has the meaning given above for formula (A-IIIb), respectively; or their pharmaceutically acceptable salts.

Particularly preferably, in any compounds having the general formulas (A-IIIb) or (A-IVb), R ⁵ means 1-3, more preferably 1 or 2 substituents, even more preferably 1 substituent, which may independently have the above meanings.

Embodiment A-4d.

A more preferred embodiment of the present invention relates to novel compounds having formulas (A-IIIb) or (A-IVb) where R ⁵ is 1 substituent, forming compounds having formula (A-IVd)

(A-IVd) where Q, R ¹ , X ¹ , X ² , X ³ , X ⁴ , R ³ , A ¹ , A ² , Y ² , R ⁴ and n are as defined above for formula (AI), (A-II), (A-IIIb) and (A-IVb), and where R ⁵ has the meaning given above for the formula (A-IIIb) and (A-IVb), respectively; or their pharmaceutically acceptable salts.

Preferably, in any of the above embodiments, one or more R ⁵ substituents are independently selected from the group consisting of the following:

a halogen atom, preferably F and Cl, optionally substituted alkyl, preferably methyl, trifluoromethyl, hydroxymethyl, hydroxy and alkoxy, preferably methoxy.

More preferably, in any of the above embodiments, one or more R ⁵ substituents are independently selected from the group consisting of the following:

a halogen atom, preferably F and Cl, and optionally substituted alkyl, preferably methyl, trifluoromethyl, hydroxymethyl.

Even more preferably, in any of the above embodiments, one or more R ⁵ substituents are independently selected from the group consisting of the following representatives: a halogen atom, preferably F and Cl, most preferably F.

Embodiment A-5.

A more preferred embodiment of the present invention relates to novel compounds having formula (A-I) and (A-II) as described above, wherein Cycl is a substituted aryl as defined above which is substituted with 1-3, preferably 1 or 2 substituents. selected from a group consisting of the following representatives:

hydroxy group, halogen atom, preferably F and Cl, preferably F, cyano group, optionally substituted alkyl, optionally substituted amino group such as (-NH2) or mono- or dialkylamino group, preferably dimethylamino group, optionally substituted acyl, optionally substituted alkoxy, preferably methoxy, difluoromethoxy and trifluoromethoxy, optionally substituted aryloxy, optionally substituted heterocyclyloxy, optionally substituted aryl, and

- 17 039916 optionally substituted heterocyclyl, preferably optionally substituted pyrrolinyl, morpholinyl and piperazinyl, optionally substituted sulfonyl group, such as preferably heterocyclyl-substituted sulfonyl, preferably having the general formula

Oh o=s=o

I such as preferably 1-3, preferably 1 or 2 substituents R ⁶ as defined above and below in the context of compounds having the general formula (A-Vb);

or their pharmaceutically acceptable salts.

Embodiment A-5b.

A more preferred embodiment of the present invention relates to novel compounds having formula (A-I) and (A-II) as described above, wherein Cycl is a substituted aryl selected from substituted phenyl, forming compounds having formula (AVb):

where Q, R ¹ , X ¹ , X ² , X ³ , X ⁴ , R ³ , A ¹ , A ² , Y ² , R ⁴ and n are as defined above for formula (AI) and (A-II) , and where R ⁶ has the above meaning, and in particular means 1-3, preferably 1 or 2 substituents, more preferably 1 substituent, selected from the group consisting of the following representatives:

hydroxy group, halogen atom, preferably F and Cl, preferably F, cyano group, optionally substituted alkyl, optionally substituted amino group such as (-NH ₂ ) or mono- or dialkylamino group, preferably dimethylamino group, optionally substituted acyl, optionally substituted alkoxy, preferably methoxy, difluoromethoxy and trifluoromethoxy, optionally substituted aryloxy, optionally substituted heterocyclyloxy, optionally substituted aryl, and optionally substituted heterocyclyl, preferably optionally substituted pyrrolinyl, morpholinyl and piperazinyl, optionally a substituted sulfonyl group, such as preferably a heterocyclyl-substituted sulfonyl, preferably having the general formula

or their pharmaceutically acceptable salts.

More preferably R ⁶ means 1 or 2 substituents, preferably R ⁶ means 1 substituent selected from the group consisting of the following representatives:

hydroxy group, halogen atom, preferably F and Cl, preferably F, cyano group, optionally substituted alkyl, optionally substituted amino group such as (-NH2) or mono- or dialkylamino group, preferably dimethylamino group, optionally substituted an alkoxy group, preferably a methoxy, difluoromethoxy and trifluoromethoxy group, an optionally substituted heterocyclyl, preferably an optionally substituted pyrrolinyl, morpholinyl and piperazinyl, and an optionally substituted sulfonyl group such as preferably a heterocyclyl-18 039916 substituted sulfonyl, preferably having the general formula

o=s=o I

Even more preferably R ⁶ is 1 substituent selected from the group consisting of the following:

a halogen atom, preferably F and Cl, preferably F, a cyano group, an optionally substituted amino group such as (-NH2) or a mono- or dialkylamino group, preferably a dimethylamino group, an optionally substituted alkoxy group, preferably methoxy, a difluoromethoxy and trifluoromethoxy group, an optionally substituted heterocyclyl, preferably an optionally substituted pyrrolinyl, morpholinyl and piperazinyl, and an optionally substituted sulfonyl group such as preferably a heterocyclyl-substituted sulfonyl, preferably having the general formula

0=3=0 I

Even more particularly preferably, in compounds having formula (AI) and corresponding to embodiments A-2, A-3, A-3b, A-4b, A-4d, A-5 and A-5b, at least one of R ¹ and R ² is a linear, branched or cyclic alkyl group substituted with a cyclic Cycl group, including the group-[CQ] _n -, where Q = H or C 1 -C ₄ -alkyl, and the resulting alkyl residue is an optionally substituted linear or a branched alkanediyl group as defined above, which is preferably selected from the following: methylene, ethane-1,2-diyl, ethane-1,1-diyl, propane-1,3-diyl, propane-1,1-diyl, propane-1,2-diyl and propane-2,2-diyl; or (in particular when Q is C1- _C4 alkyl) an optionally substituted cycloalkyl group as defined above, which is preferably selected from cyclopropane and cyclohexane;

which in a more preferred embodiment may preferably form a fused bicyclic ring with Cycl being a 6-membered heteroaryl as defined above. More preferred is an optionally substituted alkanediyl radical as defined above. Even more preferably, such an optionally substituted alkanediyl residue is selected from the group consisting of methylene, ethane-1,2-diyl, ethane-1,1-diyl and propane-2,2-diyl; more preferably methylene or ethane-1,2-diyl; most preferred is methylene.

Particularly preferred, in the compounds of the present invention, such as in particular the compounds of formulas (AI) and (A-II) and in the embodiments A-2, A-3, A-3b, A-4b described above , A-4d, A-5 and A-5b, A ¹ and A ² are each optionally substituted alkanediyl as defined above and are the same or different and independently selected from optionally substituted methylene and ethane-1,2-diyl , or A ¹ and R ³ together with the nitrogen atom to which they are attached form an optionally substituted 4-6 membered mono- or bicyclic ring, preferably a 4- or 6-membered mono- or bicyclic ring, more preferably a 4-membered ring , which is defined above.

More preferably

A ¹ and A ² are the same and are methylene,

A ¹ and A ² are the same and represent ethane-1,2-diyl,

A ¹ is methylene and A ² is ethane-1,2-diyl,

A ¹ is ethane-1,2-diyl and A ² is methylene,

A ¹ and R ³ together with the nitrogen atom to which they are attached form an optionally substituted 4-6 membered aliphatic mono- or bicyclic ring, preferably a 4-membered ring, and A ² is methylene, or

A ¹ and R ³ together with the nitrogen atom to which they are attached form an optionally substituted

- 19 039916

4-6 membered aliphatic mono- or bicyclic ring, preferably a 4-membered ring, and A ² is ethane-1,2-diyl; more preferably

A ¹ and A ² are the same and represent ethane-1,2-diyl,

A ¹ is ethane-1,2-diyl and A ² is methylene or

A ¹ and R ³ together with the nitrogen atom to which they are attached form an optionally substituted 4-membered monocyclic ring, and A ² is ethane-1,2-diyl; even more preferable

A ¹ and A ² are the same and represent ethane-1,2-diyl, or

A ¹ is ethane-1,2-diyl and A ² is methylene.

In a more preferred embodiment of the compounds of the present invention, the individual substituents have the following meanings in each case.

1. a) X ¹ is O, X ³ is N, and X ² and X ⁴ are C, and/or

b) X ¹ is N, X ³ is O, and X ² and X ⁴ are C, and/or

c) X ¹ is N, X ³ is S, and X ² and X ⁴ is C, and/or

d) X ² and X ⁴ are N and one of X ¹ and X ³ is N and the remainder is C, and/or

e) X ¹ and X ⁴ are N, X ² is C, and X ³ is O;

particularly preferred is a heterocyclic 5-membered ring selected from oxazolyl, thiazolyl, pyrazolyl, triazole, oxadiazolyl, as well as isooxazolyl and isothiazolyl.

2. Y ¹ is a hydrogen atom, optionally substituted alkyl as defined above, preferably C1, C ₂ or C ₃ alkyl as defined above, more preferably methyl.

3. n is 1.

4. Q is H.

5. Cycl is a group having the general formula

F

where * indicates the attachment point.

6. One of R ¹ and R ² is designated as R ¹ * and represents a hydrogen atom, and one of R ¹ or R ² is designated as R ² * and is selected from a hydrogen atom and optionally substituted alkyl, as defined above, preferably aryl -substituted alkyl; and heteroaryl-substituted alkyl, where the aryl and heteroaryl substituent may each carry from 1 to 3 substituents, the meaning of which is given above. Particularly preferred is the case where at least one of R ¹ or R ² which is designated R ² * is an optionally substituted aryl-methyl or an optionally substituted heteroaryl-methyl, most preferred is an optionally substituted heteroaryl-methyl and a substituted phenyl.

7. A ¹ and A ² are optionally substituted alkanediyl and are the same or different and independently selected from the following:

A ¹ and A ² are the same and are methylene,

A ¹ and A ² are the same and represent ethane-1,2-diyl,

A ¹ is methylene and A ² is ethane-1,2-diyl,

A ¹ is ethane-1,2-diyl and A ² is methylene,

A ¹ and R ³ together with the nitrogen atom to which they are attached form an optionally substituted 4-membered monocyclic ring, and A ² is methylene, or

A ¹ and R ³ together with the nitrogen atom to which they are attached form an optionally substituted 4-membered monocyclic ring, and A ² is ethane-1,2-diyl.

Particularly preferred is the case where A ¹ is methylene or ethane-1,2diyl and A ² is ethane-1,2-diyl, or when A ¹ and R ³ together with the nitrogen atom to which they are attached optionally form substituted 4-membered monocyclic ring, and A ² represents ethane-1,2-diyl.

8. R ³ is a hydrogen atom or an optionally substituted alkyl as defined above, or A ¹ and R ³ together with the nitrogen atom to which they are attached form an optionally substituted 4-membered monocyclic ring, preferably a hydrogen atom.

9. Ar may be Het-1 as defined above, and is preferably an optionally substituted mono- or bicyclic heteroaryl as defined above, preferably an optionally substituted benzimidazolyl as defined above.

In a more preferred embodiment, R ¹ and R ² are different, with one being a hydrogen atom and the other being an optionally substituted alkyl. More

- 20 039916 preferably one of R ¹ and R ² is a hydrogen atom and the other is an alkyl residue which is substituted with an optionally substituted aryl group as defined above, preferably with an optionally substituted phenyl group as defined above, or optionally substituted a heteroaryl group as defined above, preferably an optionally substituted pyridinyl group, an optionally substituted pyridazinyl group, an optionally substituted pyrimidinyl group, an optionally substituted pyrazolyl group, an optionally substituted imidazolyl group.

Even more preferably, one of R ¹ and R ² is a hydrogen atom and the other is an alkyl residue which is substituted with an optionally substituted phenyl group, an optionally substituted pyridinyl group, an optionally substituted pyridazinyl group, an optionally substituted pyrimidinyl group,

Even more preferably, one of R ¹ and R ² is a hydrogen atom and the other is an alkyl residue which is substituted with an optionally substituted phenyl group, preferably a substituted phenyl group as defined above, or an optionally substituted pyridinyl group, where optionally substituted pyridinyl a group as a substituent for an alkyl residue in one of R ¹ and R ² is most preferred.

More preferably, a halogen-substituted pyridinyl group is selected, such as, in particular, a pyridinyl group substituted with one fluorine substituent, such as, in particular, a group having the formula

Even more preferably, in the embodiments described, Ar is a bicyclic heteroaryl such as, in particular, benzimidazole, in particular benzimidazol-2-yl, having the formula

More preferably, A ¹ and A ² are each optionally substituted alkanediyl as defined above, such as very preferably A ¹ and A ² are the same and are methylene, or A ¹ and A ² are the same and are ethane-1 ,2-diyl, or A ¹ is methylene and A ² is ethane-1,2-diyl, or A ¹ is ethane-1,2-diyl and A ² is methylene, more preferably when A ¹ and A ² are the same and are ethane-1,2-diyl, or when A ¹ is ethane-1,2-diyl and A ² is methylene, or when A ¹ and R ³ are together with the nitrogen atom to which they are attached form an optionally substituted 4-membered monocyclic ring, and A ² is ethane-1,2-diyl.

Particularly preferably, the compounds according to the present invention are selected from the compounds:

- 21 039916

- 22 039916

-23039916

- 24 039916

-25 039916

- 26 039916

- 27 039916

- 28 039916

- 29 039916

- 30 039916

- 31 039916

- 32 039916

-33039916

- 34 039916

- 35 039916

- 36 039916

- 37 039916

or their pharmaceutically acceptable salts.

More preferred are compounds having formula (A-II) wherein Cycl is an optionally substituted, optionally fused heteroaryl, such as Examples No. 1, 2, 3, 4, 5, 6, 7, 8, 12, 16, 35, 36, 37, 38, 39, 40, 42, 43, 44, 45, 46, 47, 48, 49, 54, 55, 56, 57, 58, 59, 60, 61, 64, 76, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 108, 109, 110, 111, 112, 113, 114, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129,

131, 132, 134, 135, 136, 137, 138, 141, 142, 144, 145, 148, 150, 151, 152, 153, 154, 155, 156, 157, 158,159,

160, 161, 162, 163, 164, 165, 166, 167, 169, 170, 171, 173, 174, 175, 176, 177, 178, 179, 180, 181, 182,183,

184, 186, 187, 188, 189, 191, 192, 193, 194, 195, 196, 198, 199, 205, 206, 207, 208, 209, 210, 211, 212,213,

214, 215, 218, 219, 220, 223, 226, 227, 228, 230, 231, 233, 236, 239, 242, 243, 244, 247, 249, 250, 251,252,

253, 255, 256, 257, 258, 261, 264, 265, 266, 267, 268, 269, 270, 271, 272, 273, 274, 275, 276, 277, 278, 279 and 280.

More preferred are compounds having formula (A-II) wherein Cycl is an optionally substituted, optionally fused 6-membered heteroaryl such as Examples No. 1, 2, 3, 4, 5, 6, 7, 8, 12, 35 , 36, 37, 38, 39, 40, 42, 43, 44, 45, 46, 47, 48, 49, 54, 55, 56, 57, 58, 59, 61, 76, 79, 80, 81, 82 , 83, 87, 89, 90, 92, 93, 94, 96, 97, 98, 99, 101, 102, 103, 104, 105, 106, 108, 109, 110, 111, 112, 113, 114, 116 , 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 131, 132, 134, 135, 136,

137, 138, 141, 142, 144, 145, 148, 150, 151, 152, 153, 154, 155, 156, 157, 158, 159, 160, 161, 162, 163,164,

165, 166, 167, 169, 170, 171, 173, 174, 175, 176, 177, 178, 179, 180, 181, 182, 183, 184, 186.187, 188.189,

191, 192, 193, 194, 195, 196, 198, 199, 205, 206, 207, 208, 209, 210, 211, 212, 213, 214, 215, 218, 219,220,

223, 226, 227, 228, 230, 231, 233, 236, 239, 242, 243, 244, 247, 249, 250, 251, 252, 253, 255, 256, 257,258,

261, 264, 265, 266, 267, 268, 269, 270, 271, 272, 273, 274, 275, 276, 277, 278, 279 and 280.

More preferred are compounds having formula (A-II) wherein Cycl is an optionally substituted, optionally fused pyridinyl group, such as

- 38 039916 examples no. 1, 2, 3, 4, 5, 6, 7, 8, 12, 35, 36, 37, 38, 39, 40, 42, 43, 45, 47, 48, 49, 54, 55 , 56, 57, 58, 59, 76, 79,

80, 81, 82, 83, 89, 90, 92, 94, 96, 97, 98, 99, 101, 102, 103, 104, 105, 106, 108, 109, 110, 111, 112, 113, 114,

116, 117, 118, 119, 120, 121, 123, 124, 125, 126, 127, 128, 131, 132, 134, 135, 136, 137, 138, 141, 142, 144,

145, 148, 150, 151, 152, 153, 154, 156, 157, 158, 159, 160, 162, 163, 164, 165, 166, 167, 169, 170, 171, 173,

176, 177, 179, 180, 181, 184, 186, 187, 189, 191, 192, 193, 194, 195, 196, 198, 199, 205, 206, 207, 208, 209,

210, 211, 212, 213, 214, 215, 218, 219, 220, 223, 226, 227, 228, 230, 231, 233, 236, 239, 242, 243, 247, 249,

250, 251, 252, 253, 255, 256, 257, 258, 264, 265, 266, 267, 268, 269, 272, 273, 274, 275, 276, 277, 278, 279 and 280.

In addition, compounds where R1/R2 is a fluoro-substituted pyridinyl group are preferred, such as Examples No. 1, ² , 3, 4, 5, 6, 7, 8, 40, 94, 112, 113, 114, 118,

119, 120, 121, 125, 126, 127, 128, 134, 135, 148, 151, 152, 154, 157, 163, 164, 165, 166, 169, 176, 177, 179,

180, 181, 186, 193, 196, 199, 206, 208, 209, 211, 212, 213, 214, 218, 223, 226, 227, 228, 230, 231, 233, 239,

242, 243, 247, 249, 250, 251, 253, 255, 256, 257, 264, 265, 266, 267, 268, 269, 272, 273, 274, 275, 276, 277,

279 and 280.

Pharmaceutically acceptable salts of the compounds of the present invention include, for example, salts with suitable anions such as carboxylates, sulfonates, sulfates, chlorides, bromides, iodides, phosphates, tartrates, methanesulfonates, hydroxyethanesulfonates, glycinates, maleates, propionates, fumarates, toluenesulfonates, benzenesulfonates, trifluoroacetates, naphthalenedisulfonates-1,5, salicylates, benzoates, lactates, salts of malic acid, salts of 3-hydroxy-2-naphthoic acid-2, citrates and acetates.

Pharmaceutically acceptable salts of the compounds of the present invention also include, for example, salts with suitable pharmaceutically acceptable bases, such as, for example, salts with alkali or alkaline earth metal hydroxides such as NaOH, KOH, Ca(OH) ₂ , Mg(OH) ₂ etc., amine compounds such as ethylamine, diethylamine, triethylamine, ethyldiisopropylamine, ethanolamine, diethanolamine, triethanolamine, methylglucamine, dicyclohexylamine, dimethylaminoethanol, procaine, dibenzylamine, N-methylmorpholine, arginine, lysine, ethylenediamine, N-methylpiperidine, 2-amino-2-methyl-propanol-(1), 2-amino-2-methyl-propanediol-(1,3), 2-amino-2-hydroxyl-methyl-propanediol-(1,3) (TRIS) and t .d.

Depending on their structures, the compounds of the present invention may exist in stereoisomeric forms (enantiomers, diastereomers) in the presence of asymmetric carbon atoms. The present invention includes the use of enantiomers or diastereomers, as well as mixtures thereof. Enantiomerically pure forms can be obtained by standard optical resolution methods such as fractional crystallization of diastereomers obtained by reaction with optically pure compounds. Since the compounds of the present invention may exist in tautomeric forms, the present invention encompasses the use of all tautomeric forms.

The compounds of the present invention may be mixtures of the various possible isomeric forms, in particular stereoisomers such as, for example, E- and Z-, syn- and anti-, as well as optical isomers. The present invention covers E-isomers, as well as Z-isomers, as well as optical isomers and any mixtures of these isomers.

The novel compounds of the present invention may be in amorphous, crystalline or partially crystalline form, or they may exist as hydrates.

It has been surprisingly found that the new compounds having formula (A-I) and other embodiments described above work as inhibitors of ferroportin, and therefore can be used as a drug, for example, in particular, as inhibitors of ferroportin.

As already explained above, ferroportin is an iron transport protein responsible for the absorption of released iron through the intestines and its transport into the circulatory system, ensuring the supply of iron to the corresponding organs and tissues. Inactivation or suppression of ferroportin makes it impossible for iron to be exported, thereby reducing intestinal absorption of iron. In the context of the present invention, inhibition of ferroportin includes inhibition of transport of iron from cells into the circulatory system and inhibition of iron absorption in the intestine. Suppression of iron transport and/or iron return can be achieved by various mechanisms, including, for example, suppression of iron transport activity of ferroportin and corresponding suppression of iron return, initiation of internalization, degradation and/or reduction of ferroportin levels, administration of hepcidin agonists, i.e. compounds that compete with hepcidin or inhibit the binding of hepcidin to ferroportin.

The inhibition of ferroportin can be assessed by measuring the inhibition of iron transport carried out by ferroportin in the iron-induced response assay (BLAzer test), described in more detail below in the examples. In addition, ferroportin inhibition can be assessed by measuring ferroportin internalization and/or degradation in a Ferroportin Internalization and Degradation Assay (FACS) or a Ferroportin Ubiquitination and Degradation Assay, each of which is discussed in more detail. described in the examples below.

- 39 039916

In addition, ferroportin inhibition can be assessed by measuring hepcidin agonist activity, for example by assessing the ability to bind to hepcidin in the Hepcidin Internalization Assay (J774), described in more detail below in the examples. In addition, inhibition of ferroportin can be determined by confirming inhibition of hepcidin binding to ferroportin, for example, in the Biophysical Ferroportin Hepcidin Binding Assay (Hep Bind FP), described in more detail below in the examples. In addition, inhibition of ferroportin can be assessed by measuring the activity of the compound in terms of its ability to block iron export with ferroportin, for example in the iron excretion suppression test, described in more detail below in the examples.

Thus, the inhibition of ferroportin according to the present invention can, in particular, be determined by the manifestation of ferroportin-inhibiting activity in at least one of the above tests, namely:

Inhibition of ferroportin-mediated iron transport in the iron-induced response assay ( _BLAzer test): IC50 [μM] not greater than 100 (<100), preferably not greater than 50 (<50), more preferably less than 50 (<50).

Ferroportin internalization and degradation assay (FACS): EC ₅₀ value [μM] not greater than 100 (< 100), preferably not greater than 50 (< 50), more preferably less than 50 (< 50).

Analysis of ferroportin ubiquitination and degradation: Western blotting visual effect + comparable to hepcidin, +/- intermediate effect and +/+/- stronger intermediate effect, + or +/+/- effect is preferred, + effect is most preferred.

Hepcidin internalization test (J774): IC ₅₀ value not greater than 100 (<100), preferably not greater than 50 (<50), more preferably less than 50 (<50).

Biophysical test for the binding of ferroportin to hepcidin: IC ₅₀ value not greater than 100 (< 100), preferably not greater than 50 (< 50), more preferably less than 50 (< 50).

Suppression of iron excretion: IC ₅₀ value is not more than 100 (<100), preferably not more than 50 (<50), more preferably less than 50 (<50).

Ferroportin inhibition can also be determined in in vivo models, as detailed in the examples below. Suitable in vivo models may include, for example, the study of hypoferremia in intact mice by measuring the decrease in the level of iron in the blood; study of the prevention of iron absorption in anemic rats by measuring the inhibition of iron in the blood; study of the correction of hypoferremia in beta2-microglobulin-deficient mice by measuring the decrease in the level of iron in the blood; a study on the prevention of iron overload in beta2 microglobulin-deficient mice by measuring total iron in the spleen or liver; study of improvements in anemia, ineffective erythropoiesis, and iron overload in a mouse model of β-thalassemia media.

The activity of the compounds of the present invention as inhibitors of ferroportin can, in particular, be determined by the methods described below in the examples.

As already described above, the inhibition of ferroportin can be carried out, for example, by hepcidin, which is thus the most important factor in the regulation of iron absorption, inhibition of ferroportin and blocking the transfer of iron from cells into the circulatory system. The authors of the present invention have unexpectedly found that several compounds described in the present text act as hepcidin mimetics or hepcidin agonists, which is also included in the inhibition of ferroportin in the context of the present invention.

Accordingly, the compounds of the present invention can also be used to inhibit the transport of iron from cells into the circulatory system and inhibition of iron absorption in the intestine, as well as hepcidin mimetics or hepcidin agonists.

Due to the activity of the compounds of the present invention as inhibitors of ferroportin, the compounds of the present invention are also particularly well suited for use in inhibiting the transport of iron by ferroportin and, as a result, for use in the prevention and/or treatment of disorders of iron metabolism leading to an increase in iron levels, diseases associated with elevated iron levels or caused by and, increased iron absorption or iron overload, such as, in particular, tissue iron overload, diseases associated with ineffective erythropoiesis, or diseases caused by low levels of hepcidin. In addition, the compounds of the present invention are suitable for use in adjuvant therapy by limiting the amount of iron available to pathogens such as Vibrio vulnificus bacteria, thus allowing the prevention or treatment of infections caused by said pathogens.

As used herein, diseases associated with, related to, caused by, or resulting in elevated iron levels, increased iron absorption, iron overload (e.g., tissue iron overload), or ineffective erythropoiesis include thalassemia, hemoglobinopathy,

- 40 039916 such as hemoglobin E (HbE) disease, hemoglobin H (HbH) disease, hemochromatosis, hemolytic anemia such as sickle cell anemia and congenital dyserythropoietic anemia.

Diseases associated with, related to, caused by, or resulting in iron elevation, iron absorption enhancement, iron overload (e.g., tissue iron overload) also include neurodegenerative diseases such as, for example, Alzheimer's disease and Parkinson's disease, where the compounds described herein are discussed. as effective by limiting the accumulation or growth of iron in tissues or cells.

The compounds of the present invention are also suitable for use in the prevention and/or treatment of radical formation, reactive oxygen species (ROS) and oxidative stress caused by iron excess or iron overload, and in the prevention and/or treatment of damage to the heart, liver and endocrine system. caused by excess iron or iron overload, as well as for the prevention and / or treatment of inflammation initiated by excess iron or iron overload.

Diseases associated with ineffective erythropoiesis include, in particular, myelodysplastic syndrome (MDS, myelodysplasia) and polycythemia vera, as well as congenital dyserythropoietic anemia.

Other diseases, disorders, and/or disease states include iron overload caused by mutations in genes involved in regulating systemic iron storage, such as hepcidin (Hamp1), hemochromatosis protein (HFE), hemojuvelin (HJV), and transferrin receptor 2 (TFR2) such as, in particular, diseases associated with HFE and HJV gene mutations, diseases associated with chronic hemolysis, sickle cell anemia, erythrocyte membrane disorders, glucose-6-phosphate dehydrogenase deficiency (G6PD deficiency), erythropoietic porphyria, Friedreich's ataxia, as well as subtypes of iron overload such as transfusion iron overload, iron toxicity, pulmonary hemosiderosis, osteopenia, insulin resistance, African iron overload, Hallervorden-Spatz disease, hyperferritinemia, ceruloplasmin deficiency, neonatal hemochromatosis, and erythrocyte disorders including thalassemia, including alpha thalassemia , betathalassemia and deltathalassemia, sickle cell disease miyu and myelodysplastic syndrome.

Other diseases and/or disorders and/or disease states associated with elevated iron levels include, but are not limited to, diseases caused by elevated iron levels, including ataxia, Friedreich's ataxia, age-related macular degeneration, age-related cataracts, age-related retinal diseases, and neurodegenerative diseases. diseases such as pantothenate kinase-associated neurodegeneration, restless leg syndrome and Huntington's disease.

The compounds of the present invention may also be suitable for use in the prevention and treatment of diseases caused by a lack of hepcidin.

In light of the foregoing, another aspect of the present invention relates to a medicament containing one or more of the compounds described above, such as, in particular, a medicament for the prevention and treatment of any of the indications, conditions, disorders or diseases described above.

Another object of the present invention relates to pharmaceutical compositions and drugs containing one or more of the compounds of the present invention described above, and optionally one or more pharmacologically acceptable carriers and/or auxiliary compounds and/or solvents. Another subject of the present invention relates to pharmaceutical compositions and medicaments containing one or more of the compounds of the present invention described above, and, optionally, one or more additional pharmaceutically effective compounds. Said pharmaceutical compositions contain, for example, up to 99 wt.%, or up to 90 wt.%, or up to 80 wt.%, or up to 70 wt.% of the compounds of the present invention, the rest being pharmaceutically acceptable carriers and/or excipients and /or solvents and/or optional additional pharmaceutically effective compounds.

In the context of the present invention, pharmaceutically acceptable carriers, excipients or solvents are commonly used pharmaceutical carriers, excipients or solvents, including various organic or inorganic carriers and/or excipients commonly used in pharmaceuticals, in particular for the preparation of solid medical preparations. Examples include fillers (such as sucrose, starch, mannitol, sorbitol, lactose, glucose, cellulose, talc, calcium phosphate, calcium carbonate; binders such as cellulose, methylcellulose, hydroxypropylcellulose, polypropylpyrrolidone, gelatin, gum arabic, polyethylene glycol, sucrose, starch ; tablet disintegrating agents such as starch, hydrolyzed starch, carboxymethyl cellulose, carboxymethyl cellulose calcium salt, hydroxypropyl starch, sodium glycol starch, sodium bicarbonate, calcium phosphate, calcium citrate; lubricants such as magnesium stearate, talc, sodium lauryl sulfate; flavoring agents such as such as citric acid, menthol, glycine, orange powder; preservatives such as sodium benzoate, sodium bisulfite, paraben (e.g. methylaparben, ethylparaben, propylparaben, butylparaben); stabilizers such as citric acid, sodium citrate, acetic acid and 039916 thobasic carboxylic acids from the titriplex series, such as, for example, diethylenetriaminepentaacetic acid (DTPA); suspending agents such as methylcellulose, polyvinylpyrrolidone, aluminum stearate; dispersing agents; diluents such as water, organic solvents;

waxes, fats and oils such as beeswax, cocoa butter; polyethylene glycol; white vaseline, etc.

Liquid drugs such as solutions, suspensions and gels usually contain a liquid carrier such as water and/or pharmaceutically acceptable organic solvents. In addition, such liquid preparations may also contain pH adjusters, emulsifiers or dispersing agents, buffering agents, preservatives, wetting agents, gelling agents (eg methylcellulose), coloring agents and/or flavoring agents. The compositions may be isotonic, that is, they may have the same osmotic pressure as blood. The isotonicity of the compositions can be adjusted with sodium chloride and other pharmaceutically acceptable substances such as, for example, dextrose, maltose, boric acid, sodium tartrate, propylene glycol and other inorganic and organic solutes. The viscosity of liquid compositions can be adjusted with pharmaceutically acceptable thickeners such as methylcellulose. Other suitable thickeners include, for example, xanthan gum, carboxymethyl cellulose, hydroxypropyl cellulose, carbomer, and the like. The preferred thickener concentration depends on the material chosen.

Pharmaceutically acceptable preservatives may be used to increase the shelf life of liquid compositions. A suitable agent is benzyl alcohol, although a variety of preservatives may also be used, including, for example, parabens, thimerosal, chlorobutanol, and benzalkonium chloride.

The pharmaceutical compositions mentioned above can be used, for example, for intravenous, intraperitoneal, intramuscular, intravaginal, intrabuccal, transdermal, subcutaneous, mucocutaneous, oral, rectal, transdermal, topical, intradermal, intragastric administration, and are available, for example, in the form of pills, tablets, enteric-coated tablets, film-coated tablets, multilayer tablets, sustained release preparations for oral, subcutaneous or cutaneous use (in particular in the form of a patch), depot preparations, dragees, suppositories, gels, creams, syrup, granules , suppositories, emulsions, dispersions, microcapsules, micropreparations, nanopreparations, liposome preparations, capsules, enteric-coated capsules, powders, inhalation powders, microcrystalline preparations, inhalation sprays, powders, drops, nasal drops, nasal sprays, aerosols, ampoules, solutions, juices, suspensions, solutions for infusion or solutions for injection, etc.

Another object of the present invention relates to medicaments or combination preparations containing one or more compounds of the present invention and at least one additional pharmaceutically active compound, such as, in particular, a compound for the prevention and treatment of iron overload and associated symptoms, preferably iron -chelating compound, or a compound for the prevention and treatment of any of the conditions, disorders or diseases described above, such as, in particular, a pharmaceutically active compound for the prevention and treatment of thalassemia, hemochromatosis, neurodegenerative diseases (such as Alzheimer's disease or Parkinson's disease) and associated with them symptoms.

Another object of the present invention relates to the use of the compounds described above in combination therapy (fixed ratio dose or free combination of dosages for sequential administration) with one or two other active ingredients (drug compounds). Such combination therapy includes the co-administration of the compounds of the present invention and at least one additional pharmaceutically active compound (drug). Fixed-ratio combination therapy comprises administering the compounds of the present invention with at least one additional pharmaceutically active compound as a fixed-ratio dose. Combination therapy in the form of a free combination of dosages for sequential administration includes the co-administration of the compounds of the present invention and at least one additional pharmaceutically active compound at arbitrary dosages of the respective compounds, either by simultaneous administration of individual compounds, or by sequential use of individual compounds, distributed over time. The at least one additional pharmaceutically active compound (drug) includes, in particular, drugs to reduce iron overload (for example Tmprss6-ASO) or iron chelators, in particular curcumin, SSP-004184, deferythrin, deferasirox, deferoxamine and/or deferiprone, or antioxidants such as N-acetylcysteine, antidiabetic agents such as GLP-1 receptor agonists, antibiotics such as vancomycin (Van) or tobramycin, drugs for the treatment of malaria, anticancer drugs, antifungals, drugs for the treatment of neurodegenerative diseases such as Alzheimer's disease and Parkinson's disease (for example, dopamine agonists such as levodopa), antivirals such as interferon-a or ribavirin, or immunosuppressants (cyclosporine A

- 42 039916 or derivatives of cyclosporine A), iron supplements, vitamin supplements, erythrocyte stimulants, anti-inflammatory agents, antithrombotics, statins, vasoconstrictors and inotropic compounds.

Another object of the present invention relates to the use of the combinations described above for the prevention and/or treatment of diseases caused by a lack of hepcidin, or disorders of iron metabolism, such as, in particular, states of iron overload, such as, in particular, thalassemia and hemochromatosis, and other diseases, as described in this text.

Another object of the present invention concerns the use of the compounds described herein alone or as part of a combination therapy described herein, in combination with blood transfusion.

Compounds, drugs or combination preparations according to the present invention can be administered orally, parenterally, as well as intravenously.

For this purpose, the compounds of the present invention are preferably formulated into medicaments or pharmaceutical compositions in the form of pills, tablets such as enteric tablets, film-coated tablets and multilayer tablets, oral sustained release preparations, depot preparations, dragees, granules. , emulsions, dispersions, microcapsules, micropreparations, nanopreparations, liposome preparations, capsules such as enteric capsules, powders, microcrystalline preparations, epipastic preparations, drops, ampoules, solutions, suspensions, solutions for infusion or solutions for injection, or in the form preparations suitable for inhalation.

In a preferred embodiment of the present invention, the compounds are administered in the form of tablets or capsules. They may be, for example, acid-resistant forms or forms with a pH-dependent coating.

The compounds of the present invention as active ingredient can be administered, for example, as a single dose of 0.001 to 500 mg/kg of body weight, for example 1-4 times a day. However, the dosage may be increased or decreased depending on the age, body weight, condition of the patient, severity of the disease, and or route of administration.

Accordingly, another subject of the present invention relates to the compounds, medicaments, compositions and combination preparations described above for the preparation of medicaments, in particular for the prevention and treatment of any of the indications, conditions, disorders or diseases described above, in particular for oral or parenteral administration.

Another object of the present invention relates to a method for the prevention and treatment, in particular for the prevention and/or treatment of disorders of iron metabolism associated with or leading to an increase in iron levels, and in particular iron overload, diseases associated with or leading to an increase in iron levels or iron overload, disorders of iron storage associated with or leading to an increase in iron levels, and diseases associated with ineffective erythropoiesis, where this method includes administering to a patient (human or animal) in need of such treatment, the compound, drug, composition or combination preparation described above.

In this context, diseases associated with, associated with, caused by or leading to an increase in iron levels or iron overload are as defined above.

Another object of the present invention relates to the use of the compounds described above for the preparation of a medicinal product, in particular for the prevention and treatment of any of the indications, conditions, disorders or diseases described above.

Compounds of the present invention having the general structural formula (A-I) and (A-II) can generally be prepared by the methods described below and as shown in the general procedures (general schemes). Accordingly, another subject of the present invention is the process described herein for the preparation of compounds having the general formula (A-I), which comprises:

a) the reaction of compounds having the formula (a)

with compounds having formula (b) NH-R1R ² to give compounds having formula (c) o r ⁱ _x JL ^X NR ³ H

R ² (s)

b) further reaction of said compounds (c) with compounds having the formula (d) o ⁿ (d),

- 43 039916 where n = 0-7, preferably 0-5, preferably 0-1 or 2, to obtain compounds having formula (A-I);

where R ¹ , R ² , Z, A ¹ , R ³ and Ar are as defined above.

In principle, this order of steps is optimal. It is also possible to start by reacting compounds (a) with compounds (d), followed by reaction with compound (b), to give compounds having formula (A-I). Several additional intermediate steps are possible, and several intermediates are prepared as detailed in the examples below. Several intermediates are also novel compounds that fall within the scope of the present invention.

Another subject of the present invention is the process described herein for the preparation of compounds having the general formula (A-II), which includes:

a) the reaction of compounds having the formula (a)

with compounds having formula (b) NH-R1R ² to give compounds having formula (c)

b) further reaction of said compounds (c) with compounds having formula (d)

where n = 0-7, preferably 0-5, preferably 0-1 or 2, to obtain compounds having formula (A-II);

where X ¹ , Y ¹ , R ¹ , R ² , Z, A ¹ , R ³ and Ar are as defined herein.

In principle, this order of steps is optimal. It is also possible to start by reacting compounds (a) with compounds (d), followed by reaction with compound (b), to obtain compounds having formula (A-II). Several additional intermediate steps are possible, and several intermediates are prepared as detailed in the examples below. Several intermediates are also novel compounds that fall within the scope of the present invention.

General scheme 1

where Rx is an optional substituent in X4 and corresponds to Y ¹ , n is an integer as specified in the context for A ¹ .

- 44 039916

General scheme I-1

General Method 1

General Method 8

EUN, SaSO,

i) MeCN, DBU ii) Fe/AcOH

OEt

NHBoc

LiOH, THF/water

General Method 5

BUT

NHBoc

General Method 6

NHR1R2 HATU, DIPEA

DXM

where Ar has the meaning specified in the text and may correspond to Het-2.

General scheme 2

General scheme 4

and he. THF/water

NHBoc _with

General methodology

O

BUT

NHR1R2

HA;U, base

DO PD sludge and DXM sludge and THF

General methodology6

n = 1.2;

Y1 = optional substituent inX^ has the value specified in the text

Ry

1} DIPEA DOPA

2} HCI, 1,4-Duxane

General technique?

General Method 3

HCl. MeOH

General Method 1

3HQ

Ry = Ag. as indicated in those ket

i}MeCN. DBU ii} Fe/AcQH

HCO Ry

General _{Ν£Ηψ D|pEA} method 3 _m

General scheme 6

General scheme 7

- 45 039916

General scheme 8 o

Rx = substituent indicated in the text

3HCI where Rx is the optional substituent R ⁴ in Ar = Het-2.

General scheme 9

Ry = Ag

Ry = Ar has the meaning specified in the text and may correspond to Het-2.

General scheme 10

- 46 039916

General scheme 11

General scheme 12

[O], SVgS13 DBU,

General methodology 15

where Y ¹ means an optional substituent in X ⁴ , Rx means an optional substituent in phenyl,

Ry = Ar has the meaning specified in the text and may correspond to Het-2.

General scheme 13

Ry = Ar has the meaning specified in the text and may correspond to Het-2.

General scheme 14

where Rx means an optional substituent in X ¹ or X ⁴ and corresponds to Y ¹ .

- 47 039916

General scheme 15

HCI, MeOH

Ry = Ag

RyCH ₂ CI, DIPEA, DMF

Ry = Ar has the meaning specified in the text and may correspond to Het-2.

General scheme 16

Ry = Ar has the meaning specified in the text and may correspond to Het-2.

General scheme 18

- 48 039916

General scheme 19

Ry = Ar has the meaning specified in the text and may correspond to Het-2.

General scheme 20

NHR1R2

HATU, DIPEA

DM+A. ΊΤΦ

General scheme 21

i) MeCN, DBU ii) Fe/AcOH

Ry = Ar has the meaning specified in the text and may correspond to Het-2.

- 49 039916

General scheme 22

Br

OEt

Η but _? With

iBuOCOCI, TEA

ΝΗ3(βοαη), THF

NBoc_______________

HJNOC

Lawson's reagent, DXM

NBoc ________

Η.

NBoc

H

Η

EtO,C

CaCO ₃ , EUN ²

NBoc

H

NBoc

NHBoc

v) MeCN, DBU ii) Fe/AcOH

General scheme 23

1) TEA, DMF

2) Mel

General scheme 24

HCOR ₃

_NaBH4 , DIPEA

MeOH

Ar has the meaning specified in the text and may correspond to Het-2.

- 50 039916

General scheme 25

Ry = Ar has the meaning specified in the text and may correspond to Het-2.

General scheme 26

N can match Het-2, Rx = optional replace text value, and

I) MeCN, DBU ii) Fe/AcOH

General scheme 27

CO, Me. .HE

NHR1R2

HATU, DIPEA

Ar has a body indicated in phenyl.

- 51 039916

General scheme 28

EUN, CaSO ₃

HCI, MeOH

NHR1R2

HATU, base

DMF or DXM or THF

General scheme 41

General scheme 42

Intermediates Scheme A

Scheme B

Scheme C

- 52 039916

General scheme K-I

General scheme K-II about

Ry = substituent in phenyl

Examples

The present invention is illustrated in more detail in the following examples. The examples given are illustrative in nature, and a person skilled in the art can expand the described particular examples to other claimed compounds.

pharmacological tests.

1. Hepcidin internalization test (J774).

This cellular assay quantifies the binding of hepcidin to ferroportin (Fpn) by microscopic detection of the internalization of fluorescently labeled hepcidin in J774 cells. J774 is a mouse macrophage cell line that has been shown to endogenously express Fpn when incubated with iron (Knutson et al., 2005). The binding of hepcidin to Fpn triggers the internalization and degradation of hepcidin and Fpn. However, the TMR (6-carboxytetramethylrhodamine) fluorophore attached to hepcidin remains associated with the cell after degradation of the peptide backbone of hepcidin. Therefore, microscopic detection of fluorescence from cell-associated TMR serves as a measure of hepcidin binding to Fpn and internalization of hepcidin and Fpn. If binding of the TMR-hepcidin complex to Fpn is prevented, cellular TMR fluorescence remains low (Durrenberger et al., 2013). The effect of small molecule Fpn inhibitory compounds in this assay was evaluated in vitro as described below.

J774 cells, which were harvested from cultures with a confluence of about 80%, were seeded at a concentration of 8x105 cells/ml in complete medium (DMEM, 10% FBS, 1% penicillin-streptomycin) containing 200 μM Fe(III)NTA (nitrilotriacetic acid), 100 μl per well in MicroClear 96-well plates (Greiner; Cat. 655090) and grown at 37° C. with 5% CO ₂ . After overnight incubation, cells were washed 4 times with prewarmed DMEM without phenol red, 30 µl/well DMEM without phenol red was added after the final wash, and 10 µl/well serial dilutions of test compounds were added in triplicate. J774 cells were pre-incubated with test compounds at 37°C with 5% CO ₂ for 15 min, after which TMR-hepcidin was added to a final concentration of 25 nM. The cells were incubated in a total volume of 50 μl at 37°C with 5% CO ₂ for 2 h, then Hoechst 33342 dye was added at a final concentration of 0.5 μg/ml to stain the nuclei, and then incubated for 10 min at 37°C for 5 % CO ₂ Cells were washed 3 times with phosphate buffered saline (PBS) and fixed in 100 µl of 4% paraformaldehyde in PBS for 15 min at room temperature. After removal of the paraformaldehyde solution, the cells were washed 3 times with phosphate-buffered saline, leaving 100 μl per well, and the plates were covered with foil. Fluorescence patterns of TMR (530-550 nm excitation / 575-625 nm emission / 400 ms exposure time) and Hoechst 33342 (360-370 nm excitation / 420-460 nm emission / 10 ms exposure time) were recorded using a ScanR spectrophotometer (Olympus) for reading tablets, equipped with a 20x aperture lens. Four images were recorded per well covering about 1500 cells per well. The resulting images were analyzed using the ScanR image analysis software. Image analysis included nuclear detection (Hoechst 33342 fluorescence), identification of cell-associated regions, application of a virtual channel, and determination of a threshold for rolling ball background reduction, after which an average sum algorithm was applied to measure the TMR of cell-associated fluorescence as a quantitative evaluation parameter. internalized TMR hepcidin. IC ₅₀ values were calculated from the mean sum data obtained using statistical processing of log(inhibitor) vs. curves. response in Prism 5 (GraphPad Software Inc., version 5.02). For each data set, the fit of the log(inhibitor) vs. model was compared. response (three parameters) and model fit log(inhibitor) vs. the answer is variable slope (four parameters) and used the IC ₅₀ data for the preferred model. IC ₅₀ data for Fpn inhibitors tested in the hepcidin internalization test are listed in Table 1. 1. The IC ₅₀ value for unlabeled hepcidin in this test is 0.015 ± 0.011 µM.

- 53 039916

In table. 1 shows mean IC ₅₀ data for Fpn inhibitors tested in the hepcidin internalization test for several measurements.

Table 1

Comm. No. J774 IC50 (µM) Comm. No. J774 IC50 (µM) Comm. No. J774 IC50 (µM) Comm. No. J774 IC50 (µM) Comm. No. J774 IC50 (µM) one 0.012 72 1.36 114 0.118 161 0.361 210 4.913 2 0.035 74 0.21 115 >25.0 (< 50) 162 0.297 211 0.747 3 0.17 75 0.53 116 1,000 163 0.828 212 8.514 four 0.155 76 0.34 117 9.695 164 0.343 214 14.1 5 0.063 77 0.35 118 0.103 165 0.100 215 27.7 7 0.4 79 0.037 119 0.164 166 1.118 218 4.5 eight 0.24 80 0.345 120 0.034 167 0.145 219 2.43 12 0.08 81 0.42 121 0.473 169 0.750 220 0.29 16 0.90 82 0.006 122 0.026 170 0.482 223 1.9 19 10.0 83 0.096 123 0.17 171 0.026 226 0.049 21 6.7 84 0.40 124 6.332 173 0.006 227 0.130 35 0.19 85 0.029 125 1.660 174 0.141 228 0.046 36 0.25 86 0.48 126 0.096 175 1.025 230 0.14 37 0.81 87 0.19 127 0.009 176 0.957 231 5.2 38 0.03 88 0.78 128 0.005 177 4.203 233 16 39 0.07 89 0.089 129 0.353 178 3.637 236 0.15 40 0.049 90 0.025 131 0.090 179 0.216 239 0.036 42 0.60 91 2.07 132 0.580 180 30.855 242 8.92 43 0.25 92 0.83 134 0.377 181 0.135 243 0.032 44 1.33 93 0.53 135 3.407 182 0.989 244 0.090 45 0.44 94 0.012 136 >10.49 (<50) 183 0.131 247 0.082 46 0.59 95 7.23 137 0.514 184 0.063 249 0.040 47 0.72 96 2.97 138 0.179 186 0.30 250 0.014 49 30.58 97 0.27 139 4.794 187 87 251 0.062 54 0.46 98 1.85 140 3.727 188 1.16 253 0.226 55 0.015 99 2.99 141 0.167 189 0.060 256 0.081 56 0.41 100 0.46 142 21.606 191 0.33 257 0.035 57 0.10 101 0.28 144 0.012 192 13.56 258 0.152 58 0.01 102 0.058 145 0.385 193 0.287 261 0.554 59 0.05 103 2.37 148 2.085 194 0.72 265 0.070 60 2.39 104 0.90 151 0.111 195 0.21 273 0.228 61 0.56 105 0.077 152 0.004 196 1.13 274 0.145 63 0.61 106 1.52 154 0.0083 198 27.05 275 26.035 64 0.13 108 0.13 155 0.347 199 0.78 276 27.160 65 0.85 109 0.076 156 2.462 205 0.37 277 0.011 68 2.5 ON 1.699 157 0.717 206 0.18 278 0.476 69 0.26 111 0.035 158 0.047 207 0.183 279 2.009 70 0.53 112 0.378 159 0.091 208 0.012 280 50,000 71 0.24 113 >25.0 (< 50) 160 0.256 209 0.379

2. Biophysical test of ferroportin binding to hepcidin.

This biophysical test was designed to more directly confirm inhibition of hepcidin binding to ferroportin (Fpn). Incubation of TMR-hepcidin with purified human Fpn isolated from Pichia pastoris yeast cells expressing human Fpn with the FLAG terminal affinity marker (Bonaccorsi di Patti, 2014) results in an increase in the fluorescent polarization (FP) of the TMR-hepcidin ligand. Small molecule Fpn inhibitors were tested for inhibition of TMR-hepcidin binding to Fpn, detecting a dose-dependent decrease in TMR AF signal, as detailed below.

A mixture of 1.3 μM human Fpn and 30 nM TMR-hepcidin in AF assay buffer containing 50 mM Tris-HCl pH 7.3, 200 mM NaCl, 0.02% DDM, 0.1% BSA was distributed in 384- small volume round bottom black well plate (Corning, Cat. 3677) at 16 µl per well. 8 µl serial dilutions of test compounds were added in duplicate, with final concentrations of Fpn and TMR-hepcidin of 1 µM and 20 nM, respectively. The plates were incubated for 90 minutes at room temperature and parallel (S) and perpendicular (P) fluorescence was measured using a Synergy H1 Fluorescence Imaging Plate Reader Spectrophotometer (BioTek). FP values were calculated as mP using the formula below.

Fnapami - Rperpend tr= ----------- xYuO

Rparall + Rperpend

IC ₅₀ values were determined from calculated mP values as described for the internalization test

- 54 039916 hepcidin and entered in the table. 2. The IC ₅₀ value for unlabeled hepcidin in this test is 0.37 ± 0.067 µM.

In table. 2 shows the multi-measurement average (AVE) IC ₅₀ data for Fpn inhibitors tested in the hepcidin ferroportin binding biophysical assay.

table 2

Connection No. FP IC50 (µM) one 0.016 2 0.017 3 0.071 5 0.0511 7 0.18 eight 0.282 Comm. No. FP IC50 (µM) Comm. No. FP IC50 (µM) Comm. No. FP IC50 (µM) Comm. No. FP IC50 (µM) Comm. No. FP IC50 (µM) 12 0.86 81 2.974 121 5D 166 0.37 219 1.1 16 1.72 82 0.374 122 0.214 167 0.104 220 0.093 19 10.22 83 1.046 123 0.112 169 0.54 223 2,680 21 1.43 84 2.412 124 3.5 170 0.28 226 0.026 35 2.16 85 1.866 125 3.7 171 0.066 227 0.096 36 3.65 86 4.957 126 0.12 173 0.031 228 0.021 37 1.90 87 2.249 127 0.023 174 0.32 230 0.058 38 0.233 88 6.757 128 0.036 175 0.95 231 1.658 39 1.34 89 0.922 129 1.078 176 1.16 233 6.776 40 0.068 90 0.418 131 0.133 177 15.44 236 0.123 42 2.17 91 12.060 132 0.57 178 1.92 239 0.038 43 1.52 92 1.268 134 0.97 179 0.42 242 27.810 44 5.34 93 1.03 135 36.90 180 22.40 243 0.034 45 2.1 94 0.044 136 6.85 181 0.089 244 0.182 46 4.34 95 13.040 137 1.04 182 0.33 247 0.178 47 3.42 96 7.286 138 0.16 183 0.19 249 0.044 49 23.97 97 2.132 139 63.1 184 0.10 250 0.019 54 11.37 98 5.713 140 6.9 186 0.14 251 0.071 55 0.087 99 4.327 141 0.049 187 35.48 253 0.1481 56 0.566 100 1.419 142 10.5 188 0.63 256 0.046 57 0.43 101 0.315 144 0.073 189 0.047 258 0.194 58 0.076 102 0.258 145 0.35 191 0.52 257 0.038 59 0.270 103 2.525 148 7.3 192 49.26 261 0.4396 60 0.974 104 1.756 150 73 193 0.074 61 1.690 105 0.420 151 0.089 194 0.73 63 0.846 106 4.457 152 0.023 195 0.077 64 1.237 108 0.478 153 57.77 196 0.87 65 0.95 109 0.172 154 0.030 199 0.45 68 4.056 110 3.422 155 0.46 205 0.29 69 1.513 111 0.051 156 0.71 206 0.036 70 1.065 112 1.035 157 4.27 207 0.047 71 0.508 FROM 71.2 158 0.041 208 0.019 72 0.931 114 0.23 159 0.035 209 0.038 74 0.451 115 109 160 0.097 210 1.877 75 1,830 116 0.058 161 0.26 211 0.154 76 5.083 117 9.0 162 0.14 212 3.758 77 2.813 118 0.25 163 2.97 214 3.188 79 0.820 119 5.3 164 0.14 215 15.610 80 2.276 120 0.071 165 0.061 218 2.2

3. Inhibition of ferroportin-mediated iron transport in an iron-induced response assay.

Intracellular iron concentrations in this test are indirectly measured by monitoring the activity of the beta-lactamase (BLA) reporter gene hybridized to the human ferritin promoter and its associated iron controlling factor regulatory element (IRE) contained in the 5' untranslated region of the ferritin mRNA. Expression of ferroportin (Fpn) in such a cell line results in iron efflux and reduced iron levels, as reflected by reduced reporter gene activity. On the other hand, inhibition of Fpn-mediated iron efflux leads to an increase in the concentration of iron in the cell, which is detected as an increased activity of the reporter gene. Small molecule compounds that are Fpn inhibitors were tested for dose-dependent effects in an in vitro iron-induced response assay as described below.

HEK-293 cell line #354 was generated by stable integration of (i) chimeric const

- 55 039916 a human Fpn-GFP gene inserted into a doxycycline-inducible pTRE-Tight-BI plasmid derivative (Clontech, Cat. 631068), and (ii) a human ferritin promoter-BLA reporter gene construct into a HEK-293 Tet-ON Advanced derivative cell lines (Clontech). To create the ferritin-BLA reporter gene construct, a 1.4 kb fragment of the human ferritin H promoter was amplified by PCR from human genomic DNA (forward primer 5'CAGGTTTGTGAGCATCCTGAA-3'; reverse primer 5'-GGCGGCGACTAAGGAGAGG-3') and introduced before the BLA gene present in the pcDNA™6,2/cGeneBLAzer™-DEST plasmid (Invitrogen, Cat. 12578-043), thereby replacing the original CMV promoter, and placing the IRE that regulates translation of the ferritin gene approximately 170 base pairs upstream of the start codon of the reporter gene . #354 cells were harvested from cultures at approximately 80% confluence, plated at 1.8x105 cells/mL in DMEM/F12 GlutaMAX™ (Invitrogen, Cat. 31331-028) containing 10% FBS (Clontech, Cat. 631106 ), 1% penicillin-streptomycin, 200 µg/ml Hygromycin B (Invitrogen, Cat. 10687010), blasticidin 5 µg/ml (Invitrogen, Cat. R-210-01), 4 µg/ml doxycycline (Clontech, Cat. 631311 ), at a volume of 50 μl per well in 384-well plates coated with poly-d-lysine (PDL) and incubated at 37°C with 5% CO2. After overnight incubation, serial dilutions of test compounds (10 μl/well) were added in quadruplicate, and the plates were further incubated overnight at 37° C. with 5% CO ₂ . Cells were washed 3 times with Hanks' balanced salt solution (HBSS), leaving 25 μl per well. BLA activity was determined by adding 5 μl/well of the GeneBlazer CCF4-AM reagent (Invitrogen, Cat. K1085) to the cells. After incubating the plates in the dark at 18°C for 60 min, the blue and green fluorescence signals were measured on a Safire2 (Tecan) scanning spectrophotometer for reading plates with an excitation wavelength of 410 nm and emission at 458 nm (blue) and 522 nm (green ). The blue/green fluorescence intensity ratio was calculated as a measure of BLA activity, and the EC ₅₀ value was determined from the calculated blue/green fluorescence ratio as described for the hepcidin internalization assay. EC ₅₀ data for the Fpn inhibitors tested are shown in Table 1. 3. The EC ₅₀ value for hepcidin in this test is 0.096 ± 0.063 μM (n = 37).

Table 3: Mean (AVE) EC ₅₀ values for Fpn inhibitors tested in the iron-induced response assay are shown for several measurements.

Table 3

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Comm. No. BLAzer EC50 (µM) Comm. No. BLAzer EC50 (µM) Comm. No. BLAzer EC50 (µM) Comm. No. BLAzer EC50 (µM) 12 8.27 93 20.55 140 >20.00 <50 191 8.38 37 7.92 94 0.53 141 1.11 193 3.64 38 2.98 97 1.81 144 0.47 194 >3.22 <50 39 2.90 99 22.80 145 4.7 195 3.55 40 1.45 100 6.56 151 0.72 196 12.72 42 36.26 101 2.92 152 0.17 199 5.70 43 30.95 102 1.85 154 0.74 205 3.83 44 18.31 105 2.63 155 8.17 206 3.26 46 38.67 108 4.12 156 16.13 207 2.76 55 1.23 109 2.62 158 0.62 208 0.50 56 10.38 111 0.62 159 1.16 209 3.38 57 2,I 112 13.47 160 1.91 211 6.1 58 1.72 114 4.45 161 17.18 220 17.0 59 1.38 116 2.79 162 4.37 226 2.34 61 37.46 118 2.69 164 2,I 227 24.90 64 4.53 120 1.60 165 2.59 228 2.12 65 32.33 122 4.33 167 2.84 230 6.38 68 10.40 123 3.04 169 8.23 236 12.72 71 1.79 126 1.26 170 3.96 239 0.88 75 6.00 127 0.42 171 1.23 243 1.40 79 0.84 128 0.097 173 0.10 244 3.86 82 0.76 129 10.56 174 7.73 247 3.15 84 13.15 131 0.75 179 25.94 249 1.55 85 18.69 132 13.94 181 3.72 250 0.46 86 22.34 134 4.09 182 6.84 251 2.27 87 16.56 135 >20.00 <50 183 3.58 253 3.176 88 13.08 136 >20.00 <50 184 1.60 256 0.628 89 5.05 137 5.75 186 4.94 257 0.636 90 4.03 138 1.72 188 >39.07 <50 258 2.525 92 17.78 139 >20.00 <50 189 3.10 265 1.998 273 3.604 274 1.122 277 0.17

4. Analysis of the internalization and decomposition of ferroportin.

HEK-293 cell line #354 (described in Example 3) was used to measure the ability of compounds to induce internalization and degradation of ferroportin (Fpn) by fluorescence activated cell sorting (FACS). Incubation of HEK-293 #354 cells in doxycycline-containing media induced expression of the human Fpn-GFP chimeric protein on the cell surface. Data from 10 independent experiments showed that growing HEK#354 cells for 48 h in the presence of 4 μg/ml doxycycline induced an average of 42.6% ± 6.4% Fpn-GFP-positive cells. Small molecule Fpn inhibitory compounds were tested for dose-dependent effects on Fpn-GFP mean fluorescence intensity (MFI) in the HEK-293 #354 cell line, as described below.

HEK#354 cells were harvested from cultures with a confluence of about 80%, plated at a concentration of 0.6x10 ⁶ cells/ml in DMEM/F12 GlutaMAX™ medium (Invitrogen, Cat. 31331-028) containing 10% FBS (Clontech, Cat. 631106 ), 1% penicillin-streptomycin (Invitrogen, Cat. 15140-122), 200 µg/mL hygromycin B (Invitrogen, Cat. 10687-010), blasticidin 5 µg/mL, (Invitrogen, Cat. R210-01), 4 μg/ml doxycycline (Clontech, Cat. 631311), at 50 μl per well in 384-well plates (Greiner; Cat. 781091) and grown at 37°C with 5% CO2. After overnight incubation, 10 μl/well quadruple serial dilutions of test compounds were added and the plates were further incubated at 37° C. with 5% CO2 overnight. Cells were washed once with FACS buffer (phosphate buffered saline containing 1% FBS, 2 mM EDTA and 0.05% NaN ₃ ), harvested in FACS buffer with 0.5 μg/ml propidium iodide (Sigma, Cat. P4864) and analyzed on a flow cytometer (CANTO™ II, BD Biosciences) equipped with a high performance sampler. Live HEK#354 cells were isolated as a propidium iodide negative population and analyzed for FpnGFP expression. The Fpn-GFP MFI value for >2000 live cells for each compound dilution was calculated using FlowJo (Tree Star's, Oregon) and the potential of Fpn inhibitors to induce Fpn-GFP internalization and degradation was calculated as described for the hepcidin internalization assay. . EC50 values for Fpn inhibitors tested in the ferroportin internalization and degradation assay

FACS are given in Table. 4. The mean EC50 for hepcidin in this assay is 0.004±0.002 µM.

Table 4: Mean (AVE) EC50 values for Fpn inhibitors tested in the ferroportin internalization and degradation assay are shown for several measurements.

Table 4

Connection No. EC50 (µM) one 0.22 2 0.63 3 1.84 four 1.198 5 0.549 7 1.89 eight 1DZ

Comm. No. EC50 (µM) Comm. No. EC50 (µM) Comm. No. EC50 (µM) Comm. No. EC50 (µM) 40 0.81 137 11.073 171 0.571 211 4.43 55 1.029 138 0.678 171-B 0.319 213x3 HC1 4.14 58 0.387 139 >20.0 <50 173 0.071 220 3.82 82 0.689 140 >20.0 <50 174 3,960 226 0.49 94 0.22 141 0.290 175 12.452 227 1.58 109 0.885 142 39.745 176 16.985 228 0.46 111 0.075 144 0.043 179 1.207 228-B 0.22 112 3.775 145 1.245 181 0.930 230 0.95 113 41.330 148 1.050 182 23.692 231 8.33 114 2.956 151 0.523 183 1,850 236 2.16 115 38.250 152 0.071 184 1.188 239 0.32 116 0.590 154 0.130 186 5.059 243 0.51 117 >25.0 <50 155 3.954 188 35.985 244 1.69 118 4.908 156 12.110 189 0.679 247 2.18 120 0.530 157 7.862 191 2.512 249 0.95 122 3.015 158 0.325 193 3.946 250 0.60 123 4.507 159 0.757 193-B 1.391 251 1.42 126 0.757 160 1.287 194 8.050 253 1.828 127 0.081 161 5,300 195 1.459 256 0.736 128 0.006 162 1.412 196 24.845 257 0.518 129 4,464 163 7.411 199 2.966 258 1.231 131 0.194 164 3.207 205 11.115 265 1.196 132 2.148 165 0.587 206 2.072 273 1.721 134 5.194 166 >20.0 <50 207 1.608 274 0.582 135 21.210 167 1.462 208 0.15 277 0.069 136 17,860 169 4.121 209 2,440

Analysis of ubiquitination and degradation of ferroportin.

It is known that contact of cells expressing ferroportin (Fpn) with hepcidin initiates ubiquitination and subsequent internalization and degradation of Fpn (Qiao, 2012). The ability of Fpn inhibitors to induce Fpn ubiquitination and degradation was examined by immunoprecipitation assay using the J774 mouse macrophage cell line, which expresses Fpn upon iron treatment.

J774 cells (DSMZ, Cat. ACC170) were seeded at a concentration of 0.8x10 ⁶ cells/ml in 15 ml of medium (DMEM Gibco Cat. 11971-025, 10% heat-inactivated FBS Gibco Cat. 10500-064, 1% penicillin-streptomycin Gibco Cat. 15140-122) containing 200 μM Fe(III)-NTA in 10 cm Petri dishes (Greiner Cat. 664160) and incubated overnight at 37°C with 5% CO2. Cells were incubated with synthetic human hepcidin (Bachem, Cat. H-5926) or Fpn inhibitory compounds for 10 min or 120 min. Cells were washed and lysed with ice-cold lysis buffer (Pierce, Life Technoligies, Cat. 87787) containing a mixture of 1X HALT protease inhibitors (Life technologies, Cat.

- 58 039916

78429) and 10 mM iodoacetamide (Sigma, Cat. I6125) to stabilize ubiquitinated proteins. Immunoprecipitation was performed using the Pierce Classic IP Kit (Life Technologies, Cat. 26146) according to the manufacturer's protocol. Briefly, 2 mg of protein in 1.25 ml of IP lysis buffer was incubated by stirring for 1 h at 4° C. with control agarose beads to pre-purify the lysate and reduce non-specific signal. The unbound lysate was then incubated overnight with 12 μg (per reaction) of affinity-purified anti-Fpn antibody F308, which was grown against the murine Fpn amino acid 224-308 chimeric protein GST. Immune complexes were captured by dropping Pierce Protein A/G Plus Agarose beads (Life Technologies, Cat. 20423) at a rate of 14 µl per reaction, and the resulting suspension was incubated for 1.5 h at 4° C. with gentle inversion mixing. The beads were washed and the immune complexes were directly eluted with 75 μl of NuPAGE LDS SDS buffer (Life Technologies, Cat. NP0007) containing DTT (Life Technologies, Cat. NP0009).

After immunoprecipitation, samples were analyzed by Western blotting using serum containing rabbit antibodies to mouse MTP1 (Alpha Diagnostic International, Cat. MTP11-A) and monoclonal mouse antibodies to mono- and polyubiquitinated conjugates (Enzo Lifesciences, Cat. BML-PW8810) for the detection of ferroportin and ubiquitin, respectively. Horseradish peroxidase-conjugated mouse monoclonal antibodies to rabbit light chain IgG (Abcam, Cat. ab99697) and to mouse IgG H&L (Abcam, Cat. ab6789) were used as secondary antibodies.

A series of eleven Fpn inhibitors were tested by this method and compared with hepcidin. As shown in FIG. 1 and in table. 5, treatment of cells with Fpn inhibitors resulted in rapid ubiquitination within 10 min (FIG. 1 upper panel) and Fpn degradation after 2 h (FIG. 1 lower panel). The degree of Fpn degradation by Fpn inhibitors was comparable to the effect of hepcidin. However, hepcidin treatment produced a higher molecular weight ubiquitinated Fpn compared to Fpn inhibitor treatment, indicating polyubiquitination with hepcidin as opposed to monoubiquitination with the Fpn inhibitor.

Tab. 5: Summary of results for Fpn inhibitors tested in Fpn ubiquitination and degradation assay. The effect of Fpn inhibitor treatment on Fpn degradation and Fpn ubiquitination was assessed visually by Western blotting (+: comparable to hepcidin; -: no effect; +/-: moderate effect).

Table 5

Connection No. Concentration (µM) Ubiquitination Three (10 min.) Decomposition Three (120 min.) one 0.12 + + 40 1.9 + + 94 oh, s + + 111 oh, s + + 126 0.8 +/- + 127 0.1 + + 128 0.05 + + 152 0.04 + +/- 167 1.5 + + 208 0.2 + + 226 0.5 + + hepcidin 0.15 + +

Fig. 1. An Fpn inhibitor initiates ubiquitination and degradation of Fpn expressed in a mouse macrophage cell line. J774 cells were incubated overnight with Fe(III)-NTA to initiate Fpn production. Cells were then treated with 10-fold IC ₅₀ concentrations determined in the hepcidin internalization assay (see Table 1), hepcidin (hepcidin, 150 nM), or Fpn inhibitors: Illustrative Example #208 (210 nM), Illustrative Example #167 (1, 5 μM), compound No. 127 (120 nM), compound No. 152 (40 nM) for 10 or 120 min before collection and immunoprecipitation with F308 anti-Fpn antibody. Cells treated without the addition of active substances were harvested after 120 min (Control).

Immunoblotting of immunoprecipitates with anti-Fpn antibody MTP1 revealed the disappearance of ferroportin 120 min after treatment with Fpn inhibitors, to a degree comparable to that of the hepcidin-treated sample (upper panel). Rapid Fpn ubiquitination was observed 10 min after treatment of cells with Fpn inhibitors and hepcidin. Protein molecular weight standards are shown on the left in kDa.

5. Suppression of iron excretion by ferroportin inhibitors.

The activity of hepcidin and ferroportin inhibitor compounds in blocking iron export via ferroportin was tested in T47D cells (ECACC, Cat. 85102201) as described below.

Cells were plated in 24-well plates (Greiner, Cat. 662160) at 350,000 cells per well and incubated overnight with 100 μM ⁵⁸ Fe ( ⁵⁸ Fe(P)-sulfate, Vifor Pharma Batch No. ROR 3085) in medium,

- 59 039916 containing 500 μM L-ascorbic acid (Sigma Aldrich, Cat. 795437). Cells were washed once with 500 µl of iron capture buffer (IUB; PIPES 40 mM, Cat. P1851, glucose monohydrate 10 mM, Cat. 49158, sodium chloride 260 mM, Cat. 71379, potassium chloride 20 mM, Cat. P9541, magnesium sulfate 2 mM , Cat. 63138, Sigma Aldrich), then once with removal buffer (2 min incubation, BPDS 100 μM, Cat. 11890 and Na ₂ S ₂ O ₄ 500 μM, Cat. 157953, Sigma Aldrich, in IUB), and again two times IUB. Serial dilutions of hepcidin (Bachem) or ferroportin inhibitors (4-0.0064 μM, 5-fold dilution) were added in a total volume of 0.6 ml per well. Cells were incubated at 37° C. with 5% CO ₂ for 20 hours. Supernatant solutions were collected and Fe content was determined using inductively coupled plasma mass spectrometry (ICP-MS, Thermo Scientific, Element 2). Separated pellets to measure the concentration of proteins. The results were plotted as nanograms of ⁵⁸ Fe in supernatant per milligram of protein in cell lysates. Illustrative Example No. 127 suppressed iron excretion to a similar extent with the endogenous Fpn ligand hepcidin (FIG. 2).

Fig. 2. Representative example of suppression of iron excretion by hepcidin (IC ₅₀ : 0.086 μM) and illustrative example No. 127 (IC ₅₀ : 0.080 μM).

6. Hypoferremia in intact mice.

Injection of synthetic hepcidin into intact mice (wild type) resulted in a decrease in plasma iron levels (by 40-50% relative to the control group) with a maximum effect 3-4 hours after administration (Rivera, 2005; Fig. 3A). These data suggest that injectable hepcidin binds to and initiates the internalization of ferroportin (Fpn) in duodenal enterocytes and splenocytes, causing a rapid fall in plasma iron levels. Similarly, orally administered small molecule Fpn inhibitors lowered plasma iron levels in WT C57BL/6 mice in a dose-dependent manner (FIG. 3B) with comparable efficacy to hepcidin. These data indicate that intact mice can be used as a simple and reliable model for testing the acute efficacy of Fpn inhibitors in vivo.

Female C57BL/6 mice (Janvier, France) at 9 weeks of age were kept on a standard diet (Harlan Provimi Kliba 3436) and administered orally (po) with compounds or an appropriate amount of vehicle in a volume of 10 ml/kg body weight. Fpn inhibitors were formulated in 0.5% methylcellulose in water or 20% Cremophor EL/water and were orally administered to mice at 10, 30, or 100 mg/kg body weight. After 3 hours, mice were euthanized preterminally in isoflurane chambers and blood samples were taken from the retroorbital plexus. Mice were sacrificed by cervical dislocation and spleen, liver, and duodenum were harvested for further biomarker analysis. All experiments were carried out in accordance with a license confirmed by the competent veterinary services. Plasma was separated by centrifugation of blood in gel-containing microcontainers, and plasma iron levels were determined using the MULTIGENT Iron assay (Abbott Diagnostics, 6K95). There were eight mice in each group, and Bonferroni's one-way analysis of variance was used for multiple comparisons to analyze statistical differences between experimental groups. The effectiveness of some Fpn inhibitors for WT C57BL/6 mice is shown in table. 6.

Fig. 3. Decreased plasma iron levels induced by hepcidin and ferroportin inhibitor Compound 94 (illustrative example No. 94). A. Kinetics of iron levels in native C57BL/6 mice injected with synthetic hepcidin (5 mg/kg) intraperitoneally (i.p.) for the indicated time.

*-*** - shows a statistically significant decrease in the level of iron in blood plasma compared with mice that were injected with phosphate-buffered saline (PBS).

B. Plasma iron levels in native C57BL/6 mice treated with indicated amounts of hepcidin (i.p.) or Compound 94 (illustrative example No. 94).

(p.o.) for 3 hours.

Tab. 6: Efficacy of Fpn inhibitors tested in a native mouse model of hypoferremia.

Decreased plasma iron levels induced by some ferroportin inhibitors administered po. native WT C57BL/6 mice at a dosage of 10, 30 and 100 mg/kg. The relative reduction in plasma iron levels 3 hours after administration was calculated by subtracting the mean plasma iron concentrations in animals treated with the Fpn inhibitor from those in animals treated with vehicle alone. The resulting difference in mean iron values between the control group and the test compound-administered group was then divided by the mean plasma iron for the control group (which received vehicle only) and tabulated as a percentage.

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Table 6

Plasma iron reduction after 3 hours (%) Connection No. Dose 10 mg/kg Dose 30 mg/kg Dose 100 mg/kg one 0 28 51 2 9 26 fifty 12 fifteen twenty 45 39 ten twenty 35 40 ten thirty fifty 55 0 twenty 55 58 twenty thirty 40 90 0 0 40 94 thirty fifty 80 118 eight 24 49 126 7 23 62 127 17 47 54 137 -2 fourteen 25 154 13 35 56 159 four 26 60 167 19 17 34 171 ten 42 61 193 13 And 31 208 fifty 65 73 228 13 26 55 239 12 twenty 51

250 5 eighteen 40 277 6 21 54

7. Prevention of iron absorption in anemic rats.

To evaluate the in vivo efficacy of ferroportin (Fpn) inhibitors in blocking iron absorption, a series of Fpn inhibitors were tested in a rat model of anemia for iron absorption. Wistar rats (3-4 weeks old, n=5, Janvier Labs) were kept on a low iron diet (Provimi-Kliba, Cat. 2039) until their hemoglobin (Hb) reached 7-8 g/dl one day prior to administration of Fpn inhibitors. 1 hour before oral administration of 0.5 mg/kg ferrous sulfate, test compounds were orally administered as a formulation in methylcellulose solution or Cremophor. Blood samples were taken from the tail vein 1 h before iron administration (-1 h), immediately after administration of Fpn inhibitors (0 h), and after 1 h (1 h), 3 h (3 h) and sometimes up to 6 h (6 h ) after administration of test compounds. Plasma iron was measured (Abbott Diagnostics, Cat. 6K95) and suppression of plasma iron rise 3 hours after administration of test compounds was calculated as a measure of the effectiveness of Fpn inhibitors in blocking iron absorption (Table 7). As shown in FIG. 4, oral administration of an Fpn inhibitor - Illustrative Example No. 55 - at a dosage of 3, 10, or 30 mg/kg reduced plasma iron by 54, 72, and 89%, respectively, 3 hours after iron administration, compared with iron in the blood plasma of control animals before the introduction of iron, adjusted for the background concentration of iron in the blood plasma of control animals that were not administered a dose of iron.

Tab. 7: Fpn inhibitors tested in a rat model of anemia for inhibition of iron absorption. Relative suppression (%) values of the increase in plasma iron concentration are shown, corrected for the mean baseline values of plasma iron concentration in the control group, which did not receive a dose of iron, in comparison with the control groups, which received only the vehicle before the introduction of iron. Shown are the mean values for the groups (n=5) administered the indicated dosages of the Fpn inhibitor. Statistically significant (two-way ANOVA with Bonferroni test) differences between test group (test compound administered) and control group (vehicle only) are shown (***p<0.001; **p<0.01, *p<0. 05).

Fig. Fig. 4. Dose-dependent blocking of iron absorption in anemic rats by an Fpn inhibitor - illustrative example No. 55. 1 hour before the oral dose of ferrous sulfate (0.5 mg/kg), illustrative example No. blue line), 10 mg/kg (green line) or 30 mg/kg (dark blue line). Administration of Illustrative Example #55 resulted in a statistically significant (p<0.001) and dose-dependent suppression of the increase in plasma iron concentration observed 3 hours after iron administration in vehicle-only treated animals (red line). Baseline plasma iron levels are also shown for the vehicle-only and no iron-treated group (black line). For each group and time point, mean values with standard deviations are given.

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Table 7

Suppression of the increase in the concentration of iron in the blood (%) after 3 hours Comm. No. Dose 1 mg/kg Dose 3 mg/kg Dose 10 mg/kg Dose 30 mg/kg Dose 100 mg/kg one but. 2.1 42.6** 649*** but. 2 but. -3 29** 52*** but. 40 but. but. 32** 53*** 92*** 55 but. 54*** 22*** 91*** Yu9*** 58 but. but. but. 64*** 95*** 94 59*** 0 7Q*** but. but. 127 but. -eight 42*** 29*** but. 154 but. 22* 16 58*** but. 159 but. 21** 32*** 2|*** but. 167 but. _39*** _34*** 42*** but. 171 but. -3 16** 34*** but. 208 but. 59*** 86*** 109*** but.

but. = not defined.

8. Correction of hyperferremia in beta2-microglobulin deficient mice.

Mutations in genes involved in regulating systemic iron storage, such as hepcidin (Hamp1), cause iron overload in mice and humans. HFE, HJV, and TFR2 molecules on hepatocytes are required to signal adequate hepcidin production, and their deficiency leads to pathophysiologically low hepcidin concentrations and excessive iron absorption. HFE mutations are the most common cause of congenital hemochromatosis (HH) in Caucasians. HFE is a member of the MHC-class isomembrane molecule that binds to beta2 microglobulin and is involved in the transcriptional regulation of hepcidin via the bone morphogenetic protein receptor (BMPR). HFE-/- mice have reduced hepcidin levels, develop hyperferremia and high liver iron, making them a suitable animal model for studying iron overload in humans (Zhou, 1998). Beta2microglobulin (b2m-/-) deficient mice develop hyperferremia and hemochromatosis, similar to HFE-/- animals, since beta2-microglobulin is required for HFE cell surface expression and HFE function (Rothenberg and Voland, 1996). Due to the unavailability of HFE mice, b2m -/- mice were used as a model of iron overload. The pilot study confirmed that HFE-/- and b2m-/- mice have similar parameters related to iron metabolism.

Female and male homozygous b2m -/- mice were obtained from Jackson Laboratories (B6,129P2B2mtm1Unc/J, Stock Number: 002087) at 6 to 7 weeks of age and kept on a standard diet (Harlan Provimi Kliba 3436) without restriction of access to food and water . Age and gender matched WT C57BL/6 mice were obtained from Charles River. To study the acute effects of ferroportin (Fpn) inhibitors on iron overload, b2m-/- mice were administered test compounds or an appropriate amount of vehicle in a volume of 10 ml/kg body weight. The Fpn inhibitor compounds were formulated in 0.5% methylcellulose/water or 20% Cremophor EL/water, and were orally administered to mice at a dosage of 50 mg/kg body weight. WT control animals received vehicle only. After 3 hours, mice were euthanized preterminally in isoflurane chambers and blood samples were taken from the retroorbital plexus. Mice were sacrificed by cervical dislocation and spleen, liver, and duodenum were harvested for further biomarker analysis. All experiments were carried out in accordance with a license confirmed by the competent veterinary services. Plasma was separated by centrifugation of blood in gel microbags (BD Biosciences) and plasma iron levels were determined using the MULTIGENT Iron assay (Abbott Diagnostics, 6K95). There were 4-9 mice in each group, and Bonferroni's one-way analysis of variance was used for multiple comparisons to analyze statistical differences between experimental groups.

To investigate the effect of Fpn inhibitors - Illustrative Example #40 and Illustrative Example #94 - under conditions of iron overload, b2m-/- mice or WT control animals were administered Fpn inhibitors or vehicle for 3 hours. Due to its genetic deficiency, b2m-/- vehicle-only mice showed significantly higher plasma iron levels compared to WT mice (FIG. 5, group mean of 60 μM in A and 56 μM in B). Administration of b2m-/- mice of Illustrative Example #40 or Illustrative Example #94 at 50 mg/kg for 3 hours corrected elevated plasma iron levels to values observed in WT control animals. These data demonstrate the acute efficacy of small molecule inhibitors of ferroportin in a relevant disease model. Correction of plasma iron has been observed in additional studies, as summarized briefly in Table 1. eight.

Fig. 5. Complete correction of elevated plasma iron in b2m -/- mice with the introduction of ferroportin inhibitors - Illustrative example No. 40 / methylcellulose (A.) and Illustrative example No. 94 / Cremophor EL (B.) - within 3 hours.

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Tab. 8: Fpn inhibitors tested in a beta2-microglobulin deficiency mouse model to reduce elevated plasma iron.

Blood was taken 1 (#) or 3 (##) hours after oral administration of the indicated dosages of Fpn inhibitors to beta2-microglobulin deficient mice and plasma iron concentrations were determined. The relative decrease (%) in plasma iron is shown, which was calculated by subtracting the mean iron values in animals treated with the Fpn inhibitor from those found in animals treated with vehicle alone. The difference in mean iron content between the control (vehicle only administered) and test (test compounds administered) group was then divided by the mean iron content of the control group and expressed as a percentage. Values for females ($) and males (J) are given separately as a marked sex difference in efficacy was noted. Statistically significant (two-way ANOVA with Bonferroni test) differences found between test compound and control groups (vehicle alone) were shown (***p<0.001; **p<0.01; *p<0 .05).

Table 8

Decreased iron content in the blood (%)

Comm. No. Dose 20 mg/kg Dose 60 mg/kg one 9 31** 52** And 31** 59** 2 9 27 57** And 29 66** 40 ^# 9 0 13 And 35** 32** 40 ^# 9 but. ten And but. 58** 94## 9 but. 47 And but. 67 127 9 47*** 74*** And 21 83** 208 ^## 9 9 49*** And 44 67**

9. Prevention of iron overload in beta2-microglobulin deficient mice.

As a result of reduced hepcidin and increased iron absorption, beta2-microglobulin-deficient (b2m-/-) mice on a standard diet accumulate excess amounts of iron in the liver, heart, and pancreas. A pilot study showed that hepatic iron accumulation in L2m-/- mice begins at 3-4 weeks of age, and that hepatic iron levels reach 4-fold compared to wild-type (WT) mice at 6 weeks of age. In addition, switching 3-week-old b2m -/- mice to a low iron diet (LID) immediately after breastfeeding prevented iron accumulation in the liver at 6-7 weeks of age. The efficacy of Fpn inhibitors in preventing hepatic iron accumulation in b2m −/− mice was studied. 3 weeks old b2-/- mice on a low iron diet (LID) were treated with an Fpn inhibitor or vehicle alone (methylcellulose; 10 ml/kg). Mice had access to drinking water supplemented with 1 mM ⁵⁸ Fe(II) sulfate and 10 mM ascorbic acid. Administration of Fpn inhibitor or vehicle followed by access to iron-containing water was continued for 14 days. The mice were euthanized and the iron content in the liver and spleen was analyzed by ICP-OES (all iron isotopes were determined), and the liver tissues were additionally analyzed for the concentration of ⁵⁸ Fe (ICP-MS). The data summarized in Table. 9 show that oral administration of Fpn inhibitors for two weeks prevented hepatic iron accumulation in L2m-/- mice and increased spleen iron concentration, indicating inhibition of ferroportin in both the intestine and spleen.

These data demonstrate the efficacy of the small molecule inhibitor ferroportin in preventing hepatic iron accumulation in L2-/- mice, which confirms the concept in the corresponding disease model.

Tab. 9: Fpn inhibitors tested in a beta2-microglobulin deficiency mouse model to suppress hepatic iron overload.

Liver and spleen were harvested after 14 days of administration (oral; bid) of the indicated dosages of Fpn inhibitors to beta2-microglobulin deficient mice. The total concentration of iron in the liver and spleen tissues was determined using the ICP-OES method, and the concentration of ⁵⁸ Fe in the liver was determined using the ICP-MS method. Relative changes (%) in tissue iron content are shown, which were calculated by normalizing the difference between mean tissue iron values for animals treated with Fpn inhibitors and for animals treated with vehicle alone, to the mean for the control group (which were treated with Fpn inhibitors only). carrier). Values for females ($) and males (J) are given separately as a marked sex difference in efficacy was noted. Statistically significant (two-way ANOVA with Bonferroni test) differences found between test compound and control groups (vehicle alone) were shown (***p<0.001;**p<0.01;*p<0 .05). but. = not defined; n.a. = not available.

Table 9

Comm. No. Increase in the total level of iron in the spleen (%) Decreased total liver iron (%) Reducing the level of ⁵⁸ Fe in the liver (%) Dose (mg/kg) twenty 60 twenty 60 twenty 60 one 9 21 65 -one fifteen four 59 28 49 16 25 -eight 22 2 9 13 one 26 45 60** 77*** to eighteen -twenty ten 28 24 70 40 9 fifty* 85*** 32 67* 44 80* 25 24 31 59*** 53* §one*** 40 9 but. 9 but. 66 but. 67 to but. 36 but. 85** but. 95** 94 9 but. 65 but. 57 but. N.d. At but. 41 but. 79 but. N.d. 127 9 71* 51 -38 2 34 63*** to -7 -16 fifty** 65*** 71*** 73*** 208 9 56** 150*** fifteen eight 71* 87** 7 21 43 41 ₈₄ ** 58 94**

10. Improvement in anemia, ineffective erythropoiesis, and iron overload in a mouse model of β-thalassemia media.

β-thalassemia is a hereditary anemia caused by mutations in the hemoglobin β-globin gene, resulting in atypical red blood cells with a shortened lifespan. The most severe form, thalassemia major, requires blood transfusions, which lead to the development of secondary iron overload. Patients with thalassemia average have transfusion-independent anemia of moderate severity but still develop iron overload due to ineffective erythropoiesis and chronic suppression of hepcidin production.

As shown in the preceding examples, oral ferroportin (Fpn) inhibitors, like hepcidin, block ferroportin-mediated iron export from cells in vitro and, when administered to wild-type mice, temporarily lower plasma iron levels. Based on these results and published studies (Schmidt PJ, et al., Blood 2013, Guo S, et al., JCI, 2013 and Casu C. et al., Blood, 2016), Fpn inhibitors were tested for their ability to prevent accumulation iron and improve erythropoiesis in thalassemia average by limiting iron absorption and recycling iron from senescent red blood cells. The efficacy of Fpn inhibitors was investigated using a mouse model of transfusion-independent β-thalassemia. Mice with heterozygous deletions of the β1 and β2 globin genes (termed Hbb th3/+ mice) develop transfusion-independent β-thalassemia, ineffective erythropoiesis, splenomegaly, and secondary iron overload in the spleen, liver, and kidneys. Heterozygous Hbb th3/+ mice were obtained from Jackson Laboratories (B6;129P-Hbb-b1tm1Unc Hbb-b2tm1Unc/J, Stock Number: 002683) at 8-18 weeks of age and kept on a low iron diet during the experiment (Harlan Provimi Kliba 2039, 13.4 ppm Fe) without food restrictions. Hbb th3/+ mice were administered twice daily with either test compound at 20 or 60 mg/kg or methylcellulose (10 ml/kg, Sigma, Cat. 274429) as vehicle. Between these doses, mice had access to drinking water supplemented with 1 mM ⁵⁸ Fe(II) sulfate (Vifor Pharma, Batch No. ROR 3096) and 10 mM ascorbic acid (Sigma, Cat. 795437) for 6 h. The concentration of ⁵⁸ Fe(II)-sulphate added to drinking water was adjusted to replace iron intake with standard rodent chow containing 250 ppm iron. Access to water free of ⁵⁸ Fe(II) sulfate and ascorbic acid was provided during the remaining 18 hours. Administration of Fpn inhibitors or vehicle followed by access to iron-containing water was repeated for 20-46 days.

Similar to that previously shown in WT and b2m −/− mice, Fpn inhibitors given for 3 h effectively lowered plasma iron levels also in Hbb th3/+ mice (Table 10), demonstrating the ability of these small molecules to induce iron level.

Hbb th3/+ mice are anemic, with hemoglobin levels in the range of 70-80 g/L. Oral administration of Fpn inhibitors to Hbb th3/+ mice for two weeks significantly increased hemoglobin levels compared to the vehicle-only control group (Table 10). The change in hemoglobin level in the test compound group compared to the control group reached 19-22 g/l by the end of the study. Additionally, hematological parameters in the peripheral blood were also measured using an automatic blood cell analyzer. Administration of Hbb th3/+ mice with Fpn inhibitors increased the number of red blood cells, increased hematocrit, and decreased the concentration of reticulocytes and the width of distribution of erythrocytes by volume.

- 64 039916 (RDW), indicating an improvement in erythropoiesis. In addition, Hbb th3/+ mice treated with Fpn inhibitors had significantly lower blood leukocyte counts compared to controls, further demonstrating the beneficial effect of Fpn inhibitors in correcting pathological parameters in this disease model. Thus, Fpn inhibitors had a significant improvement in anemia and corrected blood composition in a mouse model of thalassemia media.

Ineffective erythropoiesis in Hbb th3/+ mice causes an overgrowth of erythroid precursors in the spleen, leading to splenomegaly. Administration of Fpn inhibitors to Hbb th3/+ mice resulted in a significant reduction in spleen weight, demonstrating the potential of Fpn inhibitors to reverse splenomegaly (Table 10).

The effect of Fpn inhibitors on erythropoiesis was investigated by analyzing the percentage of differentiating erythroid progenitors in bone marrow and spleen using flow cytometry and markers Ter119 (eBioscience, Cat. 17-5921) and CD44 (BioLegend, Cat. 103028). Bone marrow or spleen cells isolated from Hbb th3/+ mice injected with Fpn inhibitors contained a significantly lower percentage of early erythroid progenitor proerythroblasts, basophilic and prolichromatic erythroblasts, and a higher percentage of mature erythrocytes compared to control group of Hbb th3/+ mice that were injected with vehicle only (Table 10). These data showed that Fpn inhibitors reduced the severity of ineffective erythropoiesis in Hbb th3/+ mice, and these results are consistent with an improvement in hematological blood parameters.

Plasma erythropoietin levels in Hbb th3/+ mice and thalassemia patients are elevated due to a response to anemia, hypoxia, and ineffective erythropoiesis (Guo et al. JCI, 2013). Hbb th3/+ mice injected with Fpn inhibitors had significantly lower plasma erythropoietin (DuoSet ELISA R&D Systems, Cat. DY959) compared to vehicle-only controls, most likely due to partially corrected anemia and improved erythropoiesis (Table 10).

An increased level of erythropoietin in Hbb th3/+ mice caused an increase in the production of erythroferron, an erythroid-regulating hormone known to inhibit hepcidin (Kautz L. et al., Nat. Genet., 2014). Consistent with reduced plasma erythropoietin, erythroferron mRNA expression was significantly reduced in the spleen of Fpn inhibitor-treated Hbb th3/+ mice compared to vehicle-only treated mice (Table 10). Erythroferron is produced by precursors of erythrocytes that proliferate massively in the spleen of Hbb th3/+ mice due to extramedullary erythropoiesis. Thus, the effect of Fpn inhibitors on the expression of erythroferron in the spleen is through the improvement of erythropoiesis.

Increased iron requirements due to ineffective erythropoiesis and chronically low hepcidin levels in thalassemia patients cause iron accumulation in organs and related pathological manifestations such as hepatocellular carcinoma and heart failure (Rivella S. Haematologica, 2015). Excess iron is absorbed in Hbb th3/+ mice as a result of inadequately low hepcidin levels compared to high iron levels in the liver, spleen, and kidneys, and increased expression of ferroportin in the duodenum (Gardenghi S., Blood, 2007). Total liver iron and ⁵⁸ Fe in the organs of Hbb th3/+ mice treated with vehicle or Fpn inhibitors were analyzed by inductively coupled plasma emission spectrometry (ICP-OES) and inductively coupled plasma mass spectrometry (ICPMS), respectively. . ⁵⁸ Fe concentrations in the liver and spleen of Hbb th3/+ mice treated with Fpn inhibitors were significantly lower than those treated with vehicle alone, demonstrating that Fpn inhibitors prevent organ iron accumulation (Table 10).

Because Fpn inhibitors are systemically available, they are able to block iron export in all ferroportin-expressing tissues, including the duodenum, spleen, and liver. Accordingly, Fpn inhibitors are expected to prevent iron absorption from the duodenum, however, they cannot remove iron already present from the liver and spleen. Indeed, the total iron content of mice treated with Fpn inhibitor or vehicle remained unchanged (data not shown). Importantly, Fpn inhibitors significantly reduced ⁵⁸ Fe concentrations in the spleen and liver of Hbb th3/+ mice, demonstrating the ability of these small molecules to prevent iron accumulation.

In addition, reactive oxygen species (ROS) were detected in bone marrow cells using a fluorescent indicator, CM-H ₂ DCFDA (Thermo Fisher Scientific, Cat. C6827). Flow cytometric analysis showed that Fpn inhibitors significantly reduced ROS levels in mature erythroid cells compared to a vehicle-only control group of Hbb th3/+ mice (Table 10).

These data showed the ability of orally administered small molecule inhibitors of ferroportin to influence the course of the disease in improving the condition in case of anemia and ineffective erythropoiesis, as well as in alleviating splenomegaly and in preventing further accumulation of iron in the liver and spleen in a model of β-thalassemia average.

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Parameter

Decreased plasma iron with 20/60 mg/kg compound Correction of anemia on day 20-48 at 20/60 mg/kg

Increase in the number of erythrocytes at a dose of compound 20/60 mg/kg Decrease in the number of reticulocytes at a dose of compound 20/60 mg/kg Increase in hematocrit at a dose of compound 20/60 mg/kg

Reduction in RDW at 20/60 mg/kg compound dose

Decrease in the number of leukocytes at a dose of compound 20/60 mg/kg Decrease in ROS in bone marrow erythrocytes

Decrease in the relative weight of the spleen at a compound dose of 20/60 mg/kg

Decreased ⁵⁸ Fe in the spleen at a compound dose of 20/60 mg/kg Prevention of accumulation of ⁵⁸ Fe in the liver at a compound dose of 20/60 mg/kg

Decrease in the content of erythropoietin in blood plasma at a dose of the compound 20/60 mg/kg Decrease in mRNA of erythroferron in the spleen at a dose of the compound 20/60 mg/kg

Comm. 1 Conn. 2

49/66% 50/69%

6/20 g/l 3/11 g/l

4/8% 0/33%

8/39% 0/11%

0/4% 0/15%

3/16% 0/15%

32/44% 29/55%

20/45% 13/65%

23/59% 16/47%

14/48% 13/40%

12/40% 14/47%

64/78% 4/27%

82/292% 461/639%

Table 10

Comm. 40 Conn. 127

28/58% 68/81%

6/13 g/l 12/20 g/l

2/22% 0/36%

19/43% 16/61%

0/1% 3/20% n/a/n/a, 19/25%

0/36% 46/66%

n.d./n.d., n.d./ 75%

23/48% 40/61%

19/51% 43/68%

20/48% 39/59%

6/37% 32/33%

n.d./n.d., 1012/3031%

Table 10: Efficacy of ferroportin inhibitors in a mouse model of thalassemia media (Hbb th3/+ mice). These Fpn inhibitors were administered twice a day for 20 days (compounds 1 and 2), 27 days (compound 127) or 46 days (compound 40). Results are expressed as difference with control group (which received vehicle only) for hemoglobin, and as % change from control group for all other parameters reported.

Preparation of exemplary compounds.

General details of experiments.

Commercially available reagents and solvents (purity: HPLC) were used without further purification. 1H NMR spectra were recorded on a Bruker DRX 500 MHz spectrometer, Bruker DPX 250 MHz spectrometer, or Bruker Avance 400 MHz spectrometer in deuterated solvents. Chemical shift values (δ) are given in parts per million.

Compounds were purified by normal phase silica gel flash column chromatography on Biotage Isolera systems using an appropriate SNAP cartridge and gradient. Alternatively, compounds were reverse phase purified using Biotage Isolera systems using an appropriate C18 SNAP cartridge and reverse phase chromatography eluent, or preparative HPLC (if indicated).

Analytical HPLC-MS.

Method A (MET/CR/1673).

Column Supelco Ascentis Express (Part No. 53802-U) 2.1 x 30mm, 2.7µm Tempo, speakers 40°C mobile phase A, Water + 0.1% Formic acid B, Acetonitrile + 0.1% Formic acid Gradient Time (min) % organic 0 5 1.5 100 1.6 100 1.61 5 Flow rate 1 ml/min Volume of administration 3 µl Detection Signal UV 215 PDA spectrum Range: 210-420 nm, pitch: 1 nm MSD signal parameters Scan Pos (Shimadzu): 100-1000 Scan Pos (MS14): 130-850

Method B (MET/CR/1600).

Column Phenomenex Gemini-NX Cl8 (Part No. 00D-4453-B0) 2.0 xlOO mm, 3 µm column Tempo, speakers 40°C mobile phase A, 2 mM ammonium bicarbonate, buffered to pH 10 B, Acetonitrile rradient Time (min) % organic 0.00 5 5.50 100 5.90 100 5.92 5 Flow rate 0.5 ml/min Volume of administration 3 µl Detection Signal UV 215 PDA spectrum Range: 210-420 nm, pitch: 1 nm MSD signal parameters Scan Pos: 100-1000

Method C (MET/CR/1416).

Column Waters Atlantis dC18 (Part No. 186001295) 2.1 x 100 mm, 3 µm Tempo, speakers 40°C mobile phase A, Water + 0.1% Formic acid B, Acetonitrile + 0.1% Formic acid Gradient Time (min) % organic 0.00 5 5.00 100 5.40 100 5.42 5 Flow rate 0.6 ml/min Volume of administration 3 µl Detection Signal UV 215 PDA spectrum Range: 210-420 nm, pitch: 1 nm MSD signal parameters Scan Pos: 100-1000

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Method D (MET/uPLC/AB101).

Column Phenomenex Kinetix-XB C18 (Part No.00D-4498- AN) 2.1 x 100 mm, 1.7 µm Tempo, speakers 40°C mobile phase A, Water + 0.1% Formic acid B, Acetonitrile + 0.1% Formic acid Gradient Time (min) % organic 0.00 5 5.30 100 5.80 100 5.82 5 Flow rate 0.6 ml/min Volume of administration 1 µl Detection Signal UV 215 PDA spectrum Range: 200-400 nm, pitch: 1 nm MSD signal parameters ScanPos: 150-850

Method E (MET/CR/1278).

Column Waters Atlantis dC18 (Part No. 186001291) 2.1 x 50 mm, 3 µm Tempo, speakers 40°C mobile phase A, Water + 0.1% Formic acid B, Acetonitrile + 0.1% Formic acid Gradient Time (min) % organic 0.00 5 2.50 100 2.70 100 2.71 5 3.50 5 Flow rate 1 ml/min Volume of administration 3 µl Detection Signal UV 215 PDA spectrum Range: 210-420 nm, pitch: 1 nm MSD signal parameters ScanPos (Shimadzu): 100-1000 ScanPos (MS 14): 130-850

Method F - MET/CR/0990.

Column Phenomenex Gemini-NX С18 (00В-4453-В0) 2.0 x 50 mm, Zmkm Tempo, speakers 40°С mobile phase A, 2 mM ammonium bicarbonate, buffered to pH 10 B, Acetonitrile Gradient Time (min) % organic 0.00 one 1.80 100 2.10 100 2.30 one 3.50 one Flow rate 1 ml/min Volume of administration 3 µl Detection Signal UV 215 PDA spectrum Range: 210-420 nm, pitch: 1 nm MSD signal parameters ScanPos: 150-850

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Method G - MET/CR/2044.

Column Thermofisher Hypercarb™ Porous Graphitic Carbon 2.1mm x 50mm, micron Tempo, speakers 40°C mobile phase A, 25 mM ammonium acetate in water (for HPLC) pH~5 B, 25 mM ammonium acetate in acetonitrile (for HPLC) Gradient Time (min) % organic 0.00 2 four 100 5 100 6 2 6.5 2 Flow rate 0.5 ml/min Volume of administration 3 µl Detection Signal UV 215 PDA spectrum Range: 210-420 nm, pitch: 1 nm MSD signal parameters ScanPos: 150-850

Method H - METUPLCMS-A-004.

Column Acquity UPLC VEN C18 2.1 mm X 50 mm, 1.7 µm Tempo, speakers room mobile phase A, Water/acetonitrile, 9:1 + 0.1% formic acid B, Acetonitrile/water, 9:1 + 0.1% formic acid Gradient Time (min) % organic 0.00 5 1.5 100 1.7 100 1.8 5 2.0 5 Flow rate 0.7 ml/min Volume of administration 4 µl Detection Signal UV 215 PDA spectrum Range: 210-420 nm MSD signal parameters ScanPos: 150-800

Method I - METUPLCMS-A-006.

Column Acquity UPLC HSS TK 2.1mm X 100mm, 1.8um Tempo, speakers 40°C mobile phase A, Water/acetonitrile, 9:1 + 0.1% formic acid B, Acetonitrile/water, 9:1 + 0.1% formic acid Gradient Time (min) % organic 0.00 5 5.30 100 5.80 100 5.82 5 6.00 5 Flow rate 0.7 ml/min Volume of administration 4 µl Detection Signal UV 215 PDA spectrum Range: 210-420 nm MSD signal parameters ScanPos: 150-800

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Method J - METUPLCMS-A-007.

Column Acquity UPLC VEN C18 2.1 X 100 mm, 1.7 µm Tempo, speakers 40°С mobile phase A, 2 mM ammonium bicarbonate B, Acetonitrile: 2 mM ammonium bicarbonate (95:5) Gradient Time (min) % organic 0.00 5 5.30 100 5.80 100 5.82 5 6.00 5 Flow rate 0.6 ml/min Volume of administration 4 µl Detection Signal UV 215 PDA spectrum Range: 210-420 nm MSD signal parameters ScanPos: 150-800

Method K - MET/UPLCMS-A/013.

Column Acquity UPLC HSS TK 2.1 X 100 mm, 1.8 µm Tempo, speakers 40°C mobile phase A, Water + 0.1% formic acid, acetonitrile + 0.1% formic acid (90:10) B, Acetonitrile + 0.1% formic acid, water + 0.1% formic acid (90:10) Gradient Time (min) % organic 0.00 thirty 5.30 100 5.80 100 5.82 thirty 6.00 thirty Flow rate 0.6 ml/min Detection Signal UV 215 PDA spectrum Range: 210-420 nm MSD signal parameters ScanPos: 150-800

Method L - MET-THERMOMS-B-015.

Column X-bridge C-18 250 X 4.6 mm, 5 µm Tempo, speakers - Volume of introduction. 10 µl mobile phase A, 2 mM ammonium bicarbonate (pH-10) / adjusted to pH 10 with liquid NH3 B, Acetonitrile Gradient Time (min) % organic θ,θ 5 10.0 100 10.5 100 and about 5 12.0 5 Detection Signal UV 215 MSD signal parameters ScanPos: 50-1000

- 70 039916

Method M - MET/CR/1410.

Column Phenomenex Kinetex Core-Shell C18 (Part No. 00D-4601-AN) 2.1 x 50 mm, 5 µm column temp. 40°C mobile phase A, Water + 0.1% Formic acid B, Acetonitrile + 0.1% Formic acid Gradient Time (min) % organic (B) 0.00 5 1.20 100 1.30 100 1.31 5 Flow rate 1.2 ml/min Volume of administration 3 µl

Preparative HPLC - method at neutral pH.

Column Waters Sunfire C18 (Part no. 186003971) 30 x 100 mm, 10 µm Tempo, speakers Room temperature mobile phase A, water B, Acetonitrile Gradient Time (min) % organic 0 ten 2 ten 2.5 fifteen 14.5 100 15.5 100 16 ten 17 ten Flow rate 40 ml/min Volume of administration 1500 µl Detection Signal | UV 215

Preparative HPLC is a low pH (acid) method.

Column Waters Sunfire C18 (Part no. 186003971) 30 x 100 mm, 10 µm Tempo, speakers Room temperature mobile phase A, Water + 0.1% Formic acid B, Acetonitrile + 0.1% Formic acid Gradient Time (min) % organic 0 5 2 5 2.5 ten 14.5 100 15.5 100 16 5 17 5 Flow rate 40 ml/min Volume of administration 1500 µl

Detection

Signal | UV 215

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Preparative HPLC is a high pH (base) method.

Column Waters Xbridge C18 (Part no, 186003930) 30 x 100 mm, 10 µm Tempo, speakers Room temperature mobile phase A, Water + 0.2% ammonium hydroxide B, Acetonitrile + 0.2% ammonium hydroxide Gradient Time (min) % organic 0 5 2.5 5 16.05 95 18.2 95 19.1 5 twenty 5 Flow rate 40 ml/min Volume of administration 1500 µl

Detection______

Signal | UV 215

Abbreviations:

AcOH AIBN BH ₃ Boc ₂ O CaCO3 CBz CDI CHC13 DAST DBU DCC DCE DXM DIAD DIPEA DMAP DMF Et2O EtOAc EtOH h HATU Acetic acid 2,2'-azo-bis-(2-methylpropionitrile) Boran Di-tert-butyl dicarbonate Calcium carbonate Benzyloxycarbamate 1,1'-Carbonyldiimidazole Chloroform N-Ethyl-N-(trifluoro-lambda-4-sulfanyl)ethanamine 1,8-Diazabicycloundec-7-ene N,N'-dicyclohexylcarbodiimide 1,2-Dichloroethane dichloromethane Diisopropyl azodicarboxylate N,N-diisopropylethylamine N,N-dimethylpyridine-4-amine N,N-dimethylformamide diethyl ether ethyl acetate ethanol watch) 1-[bis-(Dimethylamino)methylene]-1H-1,2,3-triazolo[4,5- b]pyridinium-3-oxide Hexafluorophosphate HCl HPLC SPS K ₂ CO ₃ KO'Bu KHMDS KHSO4 LiAlH ₄ LiCl LiOH MeCN MeI MeOH min MW NaBH ₄ NaHCO ₃ NaH NaOH NBS NCS NH4Cl Hydrochloric acid High Performance Liquid Chromatography Isopropyl alcohol Potassium Carbonate Potassium Tert-Butoxide Potassium 1,1,1,3,3,3-hexamethyldisilazan-2-ide Potassium bisulfate Lithium aluminum hydride lithium chloride lithium hydroxide Acetonitrile methyl iodide methanol minute(s) Molecular weight sodium borohydride sodium bicarbonate Sodium hydride (60% in mineral oil) Sodium hydroxide N-bromosuccinimide N-chlorosuccinimide ammonium chloride

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Pd/C Palladium on charcoal PdCl ₂ (dppf) Dichloro[1,1'-bis(diphenylphosphino)ferrocene]palladium(I) Pd2dba3 tris-(Dibenzylideneacetone)dipalladium(0) PPh ₃ Triphenylphosphine PTSA p-Toluenesulfonic acid MTBE tert-Butyl methyl ether TBSCl tert-Butyldimethylsilyl chloride TEA Triethylamine TFA Trifluoroacetic acid TMOF Trimethyl orthoformate Xantphos 4,5-bis-(Diphenylphosphino)-9,9-dimethylxanthene

Intermediates

Diagram A above:

tert-butyl N-[(3-fluoropyridin-2-yl)methyl]carbamate (A1)

Suspension of 3-fluoropyridine-2-carbonitrile (8.0 g, 6.55 mmol), di-tert-butyl dicarbonate (15.7 g, 72.07 mmol), TEA (10.05 ml, 72.07 mmol) in EtOH (300 ml) was purged with nitrogen. Pd/C (10 wt%, 0.7 g, 6.55 mmol) was added and the reaction mixture was stirred under hydrogen atmosphere for 16 h. The reaction mixture was filtered through celite, washed with MeOH (100 ml) and the filtrates were evaporated in vacuo to give raw product. Purification by flash column chromatography (eluting with a 0-70% EtOAc/heptane gradient) gave the title compound (11.3 g, 72%) as an off-white solid.

¹ H NMR (DMSO-a ₆ , 250 MHz): δ [ppm] = 8.41-8.31 (m, 1H), 7.65 (ddd, J=10.1, 8.3, 1.3 Hz, 1H), 7.38 (dt, J=8.5, 4.4 Hz, 1H), 7.18 (s, 1H), 4.30 (d, J=5.4 Hz, 2H), 1.37 (s, 9H).

HPLC-MS (method A): [m/z]: 226.9 [M+H]+.

(3-Fluoropyridin-2-yl)methanamine dihydrochloride (A2) F

2HCI

Similar to general procedure 2, tert-butyl N-[(3-fluoropyridin-2-yl)methyl]carbamαt (A1) (11.3 g, 47.45 mmol) and 12M HC (59.3 ml, 711.72 mmol ) in MeOH (150 mL) at 40° C. for 2 h gave the title compound (9.7 g, 100%) as an off-white solid.

1H NMR (Methanol-J4, 500 MHz): δ [ppm] = 8.48 (dt, J=4.7, 1.3 Hz, 1H), 7.69 (ddd, J=9.7 , 8.5, 1.2 Hz, 1H), 7.50 (dt, J=8.8, 4.5 Hz, 1H), 4.37 (s, 2H).

HPLC-MS (method A): [m/z]: 126.9 [M+H] ⁺ .

Scheme B above: (4,6-dimethylpyridin-3-yl)methanamine hydrochloride (B1)

2HCI

4,6-Dimethylpyridine-3-carbonitrile (0.15 g, 1.135 mmol) in MeOH (150 ml) was placed in a HCube apparatus with 10% palladium on carbon at a flow rate of 1 ml/min using H ₂ at 50 bar at room temperature in 1M HC solution (1 ml). The solvent was evaporated in vacuo to give the title compound (190 mg, 64%) as a white solid. Used further without purification.

1H NMR (DMSO-d ₆ , 500 MHz): δ [ppm] = 8.74-8.66 (m, 1H), 8.62-8.42 (m, 3H), 7.76- 7.64 (m, 1H), 4.234.13 (m, 2H), 2.66-2.63 (m, 3H), 2.58-2.54 (m, 3H).

HPLC-MS (Method E): [m/z]: 136.9 [M+H] ⁺ .

Scheme C above: 2-(hydroxymethyl)benzonitrile (C1)

1M BH ₃ in THF (1.51 ml) was added to an ice-cold (0°C) solution of 3-formylpyridine-2-carbonitrile (200 mg, 1.51 mmol) in THF (5 ml). The reaction mixture was allowed to warm to room temperature and stirred for 15 hours. The reaction mixture was poured into ice/water (25 ml). The aqueous layer was extracted with ethyl acetate (3x20 ml). The combined organic layers were dried (Na ₂ SO ₄ ), filtered and the solvent was evaporated to give a brown oil. Purification by flash column chromatography (elution with a 20-100% EtOAc/heptane gradient) gave the title compound (45.5 mg, 22.4%) as a yellow solid.

1H NMR (CDCl ₃ , 500 MHz): δ [ppm] = 8.55 (dd, J=4.7, 1.4 Hz, 1H), 8.01-7.95 (m, 1H) , 7.49 (dd, J=8.0.4.7

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Hz, 1H), 4.89 (s, 2H).

HPLC-MS (method A): [m/z]: 134.85 [M+H]+.

2-{[(tert-Butyldimethylsilyl)oxy]methyl}benzonitrile (C2)

1M TBSCl in DCM (0.369 ml, 0.369 mmol) was added dropwise to a solution of 3(hydroxymethyl)pyridine-2-carbonitrile (C1) (45 mg, 0.335 mmol) and imidazole (46 mg, 0.671 mmol) in DMF (2 ml) . The reaction mixture was stirred at room temperature for 15 hours. The solvent was evaporated and the crude product was purified by flash column chromatography (eluting with a gradient of 0-50% EtOAc-heptane) to give the title compound (44 mg, 52.8%) in yellow oil.

1H NMR (CDCl ₃ , 500 MHz): δ [ppm] = 8.60-8.58 (m, 1H), 8.10-7.96 (m, 1H), 7.53 (dd, J=8.0, 4.7 Hz, 1H), 4.94(s, 2H), 0.95(s, 9H), 0.15(s, 6H).

HPLC-MS (method A): [m/z]: 249.00 [M+H] ⁺ .

(3-{[(tert-Butyldimethylsilyl)oxy]methyl}pyridin-2-yl)methanamine (C3)

2M LiAlH4 in THF (0.09 mL) was added dropwise to an ice-cold solution (0°C) of 3-{[(tert-butyldimethylsilyl)oxy]methyl}pyridine-2-carbonitrile (C2) (44 mg, 0.18 mmol) in THF (3 ml). The reaction mixture was allowed to warm to room temperature and stirred for 2 h. Diethyl ether (5 ml) was added followed by H ₂ O (1 ml) followed by 20% w/w NaOH solution (1 ml) and water (3 ml). Separate the layers. The aqueous layer was extracted with ethyl acetate (3x10 ml). The combined organic layers were dried (Na ₂ SO ₄ ), filtered and the solvent was evaporated. The crude product was purified by flash column chromatography (eluting with a 0-100% EtOAc/heptane gradient) to give the title compound (10 mg, 22.4%) as a yellow oil.

HPLC-MS (Method A): [m/z]: 252.95 [M+H]+.

N-(2-nitrophenyl)prop-2-enamide (D)

_{Acryloyl chloride} (11.8 ml , 145 mmol) and the resulting mixture was stirred for 16 h. The reaction mixture was filtered and evaporated in vacuo to give the crude product. Purification by flash column chromatography (eluting with a 10-15% EtOAc/heptane gradient) gave the title compound (6.95 g, 78%) as a yellow solid.

¹ H NMR (CDCl ₃ , 250 MHz): δ [ppm] = 10.59 (s, 1H), 8.90 (dd, J=8.6, 1.3 Hz, 1H), 8, 25 (dd, J=8.5, 1.6 Hz, 1H), 7.68 (dd, J=8.5, 7.3, 1.4 Hz, 1H), 7.21 (dd, J= 8.6, 7.3, 1.4 Hz, 1H), 6.54-6.28 (m, 2H), 5.89 (dd, J=9.9, 1.3 Hz, 1H).

HPLC-MS (method A): [m/z]: 192.9 [M+H] ⁺ .

2-(Chloromethyl)-5-(trifluoromethyl)- 1H-1,3-benzodiazol (E)

M HC (1 ml, 12 mmol) was added to a mixture of 4-(trifluoromethyl)benzene-1,2-diamine (1 g, 5.68 mmol) and chloroacetic acid (0.590 g, 6.25 mmol) in water (20 ml ) and the resulting mixture was heated at 100° C. for 2 hours. An additional 12M HC (4 ml, 48 mmol) was added and the reaction mixture was heated at 120° C. for 3 hours. The resulting mixture was then cooled to room temperature and quenched by adding 7M ammonia in MeOH until basic, was extracted with ethyl acetate (3x20 ml), and the combined organic layers were washed with saturated sodium chloride solution (20 ml), dried (MgSO ₄ ), filtered and evaporated in vacuum. Flash column chromatography (elution with a gradient of 5-50%

- 74 039916

EtOAc/heptane) gave the crude title compound as a purple solid (0.571 g, 24%, 56% pure) which was used without further purification.

HPLC-MS (Method E): [m/z]: 234.85 [M+H]+.

tert-Butyl 2-(chloromethyl)methyl-1H-1,3-benzodiazol-1-carboxylate (F)

To a solution of 2-(chloromethyl)-6-methyl-1H-1,3-benzodiazol (1 g, 6 mmol) in DMF (20 ml) was added DIPEA (1.4 g, 11 mmol), then Boc anhydride (1. 8 g, 8 mmol). The reaction mixture was stirred for 18 hours. Water was added to the reaction mixture and extracted with ethyl acetate. The organic phase was dried with Na ₂ SO ₄ , evaporated in vacuo to give crude product which was purified by flash column chromatography eluting with n-hexane to ethyl acetate/n-hexane (5:95) to give the desired product in as a yellow oil (0.7 g, 22%). The target product was obtained as an inseparable mixture and used in the next step.

¹ H NMR (CDCla, 400 MHz): δ [ppm] = 7.84 (d, J=8.7 Hz, 2H), 7.62 (d, J=8.2 Hz, 1H), 7.53 (s, 1H), 7.20 (dd, J=13.0, 4.6 Hz, 2H), 5.05 (s, 2H), 5.04 (s, 2H), 2.50 (s, 3H), 2.47 (s, 3H), 1.74 (s, 9H), 1.73 (s, 9H).

N-(3-fluoro-2-nitrophenyl)-prop-2-enamide (G) o HN'^>

Ij

Prop- ₂ - _enoyl chloride (1.0 ml, 12.8 mmol). The reaction mixture was allowed to stir at room temperature for 16 hours. The reaction mixture was filtered, evaporated in vacuo and purified by flash column chromatography (eluting with a 0-70% EtOAc/heptane gradient) to give the title compound (604 mg, 87% ) as a yellow solid.

1H NMR (DMSO-d6, 250 MHz): δ [ppm] = 10.58 (s, 1H), 7.69 (m, 1H), 7.46-7.33 (m, 2H), 6.43 (dd, J=17.0, 9.8 Hz, 1H), 6.27 (dd, J=17.0, 2.1 Hz, 1H), 5.85 (dd, J=9, 8, 2.1 Hz, 1H).

HPLC-MS (method A): [m/z]: 210.95 [M+H] ⁺ .

N-(3-chloro-2-nitrophenyl)-prop-2-enamide (H)

Acryloyl chloride (1.03 ml, 12.67 mmol) was slowly added to a suspension of 3-chloro-2-nitroaniline (0.729 g, 4.22 mmol) and K ₂ CO ₃ (2.34 g, 16.9 mmol) in acetone (20 ml). The reaction mixture was stirred at room temperature for 4 hours, filtered and the resulting residue was washed with acetone. The combined filtrates were evaporated in vacuo. Purification by flash column chromatography (eluting with a 0-60% EtOAc/heptane gradient) gave the title compound (0.52 g, 47%) as a yellow solid.

1H NMR (CDCl3, 500 MHz): δ [ppm] = 8.36 (dd, J=8.3, 1.1 Hz, 1H), 8.28 (s, 1H), 7.49 ( dd, J=8.3, 8.3 Hz, 1H), 7.32 (dd, J=8.3, 1.1 Hz, 1H), 6.47 (dd, J=16.9, 0, 8 Hz, 1H), 6.25 (dd, J=16.9, 10.3 Hz, 1H), 5.90 (dd, J=10.3, 0.8 Hz, 1H).

HPLC-MS (method M): [m/z]: 227.00 [M+H] ⁺ .

N-(2-methoxy-6-nitrophenyl)-prop-2-enamide (I)

Acryloyl was added dropwise to a nitrogen-purged and stirred suspension of 2-methoxy-6-nitroaniline (0.52 g, 3.09 mmol) and K ₂ CO ₃ (1.71 g, 12.4 mmol) in acetone (30 ml). chloride (0.754 ml, 9.28 mmol). The reaction mixture was allowed to stir at room temperature for 16 hours. The resulting mixture was filtered, evaporated, diluted with EtOAc, washed with water, dried (MgSO ₄ ), filtered and evaporated to give the crude product. Purification by flash column chromatography (eluting with a gradient of 0-100% EtOAc/heptane followed by 0-2% MeOH/EtOAc) gave the title compound (0.674 g, 96%) as an orange solid.

1H NMR (CDCl3, 250 MHz): δ [ppm] = 7.82 (s, 1H), 7.57 (dd, J=8.2, 1.3 Hz, 1H), 7.31 ( t, J=8.3 Hz, 1H), 7.19 (dd, J=8.3, 1.3 Hz, 1H), 6.47 (dd, J=17.0, 1.7 Hz, 1H ), 6.33 (dd, J=17.0, 9.8 Hz, 1H), 5.85 (dd, J=9.8, 1.7

- 75 039916

Hz, 1H), 3.97 (s, 3H).

HPLC-MS (method M): [m/z]: 223.05 [M+H]+.

N-(5-fluoro-2-nitrophenyl) prop-2-enamide (J) o

Acryloyl chloride (3.8 ml, 46.5 mmol) was slowly added to a suspension of 5-fluoro-2-nitroaniline (2.4 g, 15.5 mmol) and K ₂ CO ₃ (8.57 g, 62 mmol) in acetone (100 mL) and the resulting mixture was stirred at room temperature for 3 days and refluxed for 6 hours. More acryloyl chloride (3.8 mL, 46.5 mmol) and DMAP (0.95 g, 7.75 mmol) were added. ), and the resulting mixture was heated at reflux for another 2 hours. The reaction mixture was cooled to room temperature and filtered. The resulting residue was washed with acetone and the combined filtrates were evaporated in vacuo. The resulting residue was redissolved in Et ₂ O (350 ml) and saturated aqueous NaHCO ₃ (200 ml). The resulting mixture was intensively stirred for 15 min. The phases were separated and the organic phase was washed with additional saturated aqueous NaHCO ₃ (100 ml) and saturated sodium chloride solution (100 ml), dried (sodium sulfate) and evaporated in vacuo. Purification by flash chromatography through a pad of silica gel (eluting with a 0-4% Et ₂ O/heptane gradient) gave the title compound (1.04 g, 32%) as a pale yellow solid.

1H NMR (CDC1 ₃ , 250 MHz): δ [ppm] = 10.83 (s, 1H), 8.79 (dd, J=11.2, 2.5 Hz, 1H), 8.34 (dd, J=9.2, 5.7 Hz, 1H), 6.99-6.82 (m, 1H), 6.53 (d, J=16.9 Hz, 1H), 6.35 ( dd, J=17.1, 9.9 Hz, 1H), 5.95 (d, J=10.1 Hz, 1H).

HPLC-MS (method M): [m/z]: 211.15 [M+H] ⁺ .

The K-I diagram above:

N-(2-chloro-5-fluorophenyl)prop-2-enamide (K1) o

Ta

Prop-2 -enoyl chloride (5.0 mL, 61.8 mmol) and stirred for 16 h. The reaction mixture was filtered, evaporated in vacuo and purified by flash column chromatography (eluting with a 0-35% EtOAc/heptane gradient) to give the title the title compound (3.99 g, 84%) as a white solid.

¹ H NMR (CDCl·,, 250 MHz): δ [ppm] = 8.40 (dd, J=10.9, 3.0 Hz, 1H), 7.79 (s, 1H), 7 .35 (dd, J=8.9, 5.6 Hz, 1H), 6.81 (dd, J=8.9, 7.6, 3.0 Hz, 1H), 6.50 (dd, J =16.9, 1.2 Hz, 1H), 6.32 (dd, J=16.9, 10.0 Hz, 1H), 5.88 (dd, J=10.0, 1.2 Hz, 1H).

HPLC-MS (Method A): [m/z]: 200.10 [M+H] ⁺ .

N-(6-chloro-3-fluoro-2-nitrophenyl)prop-2-enamide (K2) o

ΗνΆ^

Into a nitrogen-purged solution of N-(2-chloro-5-fluorophenyl)prop-2-enamide (K1) (3.99 g, 17.4 mmol), conc. H ₂ SO ₄ (15 ml) and AcOH (6 ml) at 0° C. was added dropwise red fuming HNO3 (1.8 ml, 38.3 mmol) and the reaction mixture was left stirring for 2 h. ice and water and was extracted with dichloromethane (4x40 ml). The combined organic extracts were dried (MgSO ₄ ), filtered, evaporated in vacuo and purified by flash column chromatography (eluting with a 0-70% EtOAc/heptane gradient) to give the title compound (1.08 g, 20%) as white solid.

1H NMR (CDC13, 250 MHz): δ [ppm] = 7.64 (dd, J=9.1, 5.0 Hz, 1H), 7.51 (s, 1H), 7.19 ( m, 1H), 6.52 (dd, J=16.9, 1.1 Hz, 1H), 6.32 (dd, J=16.9, 10.2 Hz, 1H), 5.94 (dd , J=10.1, 1.1 Hz, 1H).

HPLC-MS (method A): [m/z]: 244.95 [M+H] ⁺ .

N-(2,4-difluorophenyl)prop-2-enamide (K3)

Prop-2-enoyl chloride ( 3.7 ml, 46.5 mmol). The reaction mixture was allowed to stir for 16 h. The reaction mixture was filtered, evaporated, purified by flash column chromatography (elution with a gradient of 0-30%

EtOAc/heptane) and triturated in heptane to give the title compound (2.9 g, 100%) as a white solid.

¹ H NMR (CDCl3, 250 MHz): δ [ppm] = 8.49-8.29 (m, 1H), 7.33 (s, 1H), 6.99-6.84 (m, 2H), 6.48 (dd, J=16.9, 1.4 Hz, 1H), 6.29 (dd, J=16.8, 10.1 Hz, 1H), 5.85 (dd, J =10.1, 1.4 Hz, 1H).

HPLC-MS (Method A): [m/z]: 183.95 [M+H]+.

N-(2,4-difluoro-6-nitrophenyl)prop-2-enamide (K4)

Into a nitrogen-purged solution of N-(2,4-difluorophenyl)prop-2-enamide (K3) (2.9 g, 15.4 mmol), AcOH (5 mL), and conc. H ₂ SO ₄ (13 ml) at 0°C was added dropwise red fuming nitric acid (1.6 ml). The reaction mixture was allowed to stir for 2 hours. The reaction mixture was poured into a mixture of ice and water and extracted with dichloromethane (4x40 ml). The combined organic extracts were washed with brine, dried (MgSO ₄ ), filtered, evaporated in vacuo and triturated in heptane to give the crude product as a beige solid (3.23 g). Purification by flash column chromatography (eluting with a 0-40% EtOAc/heptane gradient) gave the title compound (1.25 g, 35.5%) as a white solid.

1H NMR (CDCl ₃ , 250 MHz): δ [ppm] = 8.17 (s, 1H), 7.67 (dt, J=7.9, 2.4 Hz, 1H), 7.34 -7.28 (m, 1H), 6.51 (dd, J=17.0, 1.4Hz, 1H), 6.35 (dd, J=17.0, 9.9Hz, 1H), 5.92 (dd, J=9.9, 1.3 Hz, 1H).

HPLC-MS (method A): [m/z]: 229.05 [M+H] ⁺ .

N-(2,5-difluorophenyl)prop-2-enamide (K5)

Prop-2 -enoyl chloride (5.0 ml, 61.96 mmol). The reaction mixture was allowed to stir at room temperature for 2 hours. The reaction mixture was filtered and the filtrate was evaporated to give a white solid which was triturated in heptane to give the title compound (2.91 g, quant) as a white solid .

1H NMR (CDCl3, 250 MHz): δ [ppm] = 8.30 (m, 1H), 7.50 (s, 1H), 7.07 (m, 1H), 6.85-6, 70 (m, 1H), 6.50 (dd, J=16.8, 1.2 Hz, 1H), 6.30 (dd, J=16.9, 10.1 Hz, 1H), 5.87 (dd, J=10.1, 1.2 Hz, 1H).

HPLC-MS (method A): [m/z]: 183.95 [M+H] ⁺ .

N-(3,6-difluoro-2-nitrophenyl)prop-2-enamide (K6)

Into a nitrogen-purged and stirred solution of N-(2,5-difluorophenyl)prop-2-enamide (K5) (2.91 g, 15.9 mmol), AcOH (5 mL), and conc. H ₂ SO ₄ (13 ml) at 0°C was added dropwise red fuming HNO ₃ (1.6 ml, 34.0 mmol). The reaction mixture was allowed to stir for 2 hours. The reaction mixture was poured into a mixture of ice and water and extracted with dichloromethane (4x40 ml). The combined organic extracts were washed with brine, dried (MgSO4), filtered and evaporated in vacuo. Purification by flash column chromatography (elution with 0-60% EtOAc/heptane gradient), followed by flash column chromatography (elution with 20% EtOAc/heptane gradient) gave the title compound (0.316 g, 8%) as white solid.

1H NMR (CDCl ₃ , 500 MHz): δ [ppm] = 7.64 (s, 1H), 7.39 (m, 1H), 7.19 (m, 1H), 6.51 (dd , J=17.0, 0.7 Hz, 1H), 6.32 (dd, J=17.0, 10.4 Hz, 1H), 5.93 (dd, J=10.4, 0.7 Hz, 1H).

HPLC-MS (method A): [m/z]: 228.95 [M+H] ⁺ .

N-[2-(trifluoromethyl)phenyl]prop-2-enamide (K7)

_Prop - _2- enoyl chloride (8.0 ml, 99.30 mmol). The reaction mixture was allowed to stir at room temperature for 3 hours. The reaction mixture was filtered, evaporated in vacuo and triturated with heptane to give the title compound (4.74 g, 86%) as a white solid.

Ή NMR (CDCl ₃ , 250 MHz): δ [ppm] = 8.34 (d, J=8.2 Hz, 1H), 7.70-7.45 (m, 3H), 7.28 -7.22 (m, 1H), 6.46 (dd, J=16.9, 1.3 Hz, 1H), 6.29 (dd, J=16.9, 10.0 Hz, 1H), 5.86 (dd, J=10.0, 1.3 Hz, 1H).

HPLC-MS (method A): [m/z]: 215.90 [M+H] ⁺ .

N-[2-nitro-6-(trifluoromethyl)phenyl]prop-2-enamide (K8)

Into a nitrogen purged solution of N-[2-(trifluoromethyl)phenyl]prop-2-enamide (K7) (4.64 g, 20.91 mmol), AcOH (5 ml) and conc. H ₂ SO ₄ (13 mL) red fuming HNO ₃ (1.6 mL, 34.05 mmol) was added dropwise at 0°C. The reaction mixture was allowed to stir at room temperature for 16 hours. The reaction mixture was poured into a mixture of ice and water and extracted with dichloromethane (4x40 ml). The combined organic extracts were dried (MgSO ₄ ), filtered and evaporated in vacuo. Purification by flash column chromatography (eluting with a 0-20% EtOAc/heptane gradient) gave the title compound (0.829 g, 12%) as a beige solid.

HPLC-MS (method A): [m/z]: 260.95 [M+H]+.

N-(2,3-difluorophenyl)prop-2-enamide (K9)

Prop-2-enoyl chloride (10 ml , 124 mmol). The reaction mixture was allowed to stir for 16 hours. The reaction mixture was filtered and the filtrate was evaporated to give a white solid which was triturated with heptane to give the title compound (4.97 g, 87%) as a white solid.

Ή NMR (CDCl ₃ , 250 MHz): δ [ppm] = 8.29-8.12 (m, 1H), 7.46 (s, 1H), 7.18-7.04 (m, 1H), 7.02-6.85 (m, 1H), 6.50 (dd, J=16.8, 1.3 Hz, 1H), 6.31 (dd, J=16.9, 10, 1 Hz, 1H), 5.87 (dd, J=10.1, 1.3 Hz, 1H).

HPLC-MS (method A): [m/z]: 184.2 [M+H] ⁺ .

N-(2,3-difluoro-6-nitrophenyl)prop-2-enamide (K10)

Into a nitrogen-purged solution of N-(2,3-difluorophenyl)prop-2-enamide (K9) (4.9 g, 26.8 mmol), AcOH (5 mL), and conc. H2SO4 (13 ml) at 0°C was added nitric acid (1.6 ml) dropwise. The reaction mixture was allowed to stir for 2 hours. The reaction mixture was poured into a mixture of ice and water and was extracted with dichloromethane (5x30 ml). The combined organic extracts were washed with saturated sodium chloride solution (50 ml), dried over MgSO ₄ , filtered and evaporated to give the crude product. It was triturated in heptane (100 ml). The resulting suspension was filtered and the resulting residue was separated to give a mixture of para/ortho-nitrated regioisomers as a beige solid (6 g). Purification by acidic prep-HPLC gave the title compound (4.2 g) as a white solid.

HPLC-MS (method A): [m/z]: 228.95 [M+H] ⁺ .

The above general scheme of K-II:

N-(4-cyanophenol)prop-2-enamide (K11)

Acryloyl chloride (0.69 ml, 8.46 mmol) was added to an ice-cold suspension of 4-aminobenzonitrile (250 mg, 2.12 mmol) and K ₂ CO ₃ (880 mg, 6.35 mmol) in acetone (5 ml). The resulting mixture was stirred for 18 hours while warming to room temperature. The reaction mixture was filtered and the resulting residue was washed with acetone (5 ml). The combined filtrates were evaporated in vacuo and the resulting residue was purified by flash column chromatography eluting with a gradient of 0-80% EtOAc/heptane,

- 78 039916 obtaining the title compound (353 mg, 96%) as a white solid.

1H NMR (CDCl ₃ , 250 MHz): δ [ppm] = 7.73 (d, J=8.8 Hz, 2H), 7.68-7.58 (m, 2H), 7.37 (s, 1H), 6.49 (dd, J=16.8, 1.0 Hz, 1H), 6.25 (dd, J=16.8, 10.2 Hz, 1H), 5.86 ( dd, J=10.2, 1.0 Hz, 1H).

HPLC-MS (method M): [m/z]: 173.45 [M+H]+.

N-(4-cyano-2-nitrophenyl)prop-2-enamide (K12)

Nitric acid (0.6 ml) was added dropwise to an ice-cold solution of N-(4-cyanophenyl)prop-2enamide (K11) (1.03 g, 5.75 mmol) in acetic acid (2 ml) and sulfuric acid (4 .75 ml). The reaction mixture was stirred for 3 h, then poured into a mixture of ice and water and was extracted with dichloromethane (4x20 ml). The combined organic extracts were dried (MgSO ₄ ) and evaporated in vacuo. Purification by flash column chromatography eluting with a 0-90% EtOAc/heptane gradient gave the title compound (1.2 g, 93%) as a yellow solid.

1H NMR (CDCl ₃ , 250 MHz): δ [ppm] = 10.77 (s, 1H), 9.14 (d, J=8.9 Hz, 1H), 8.58 (d, J =2.0 Hz, 1H), 7.90 (dd, J=8.9, 1.7 Hz, 1H), 6.54 (dd, J=17.0, 0.9 Hz, 1H), 6 .35 (dd, J=17.0, 10.1 Hz, 1H), 5.98 (dd, J=10.1, 0.9 Hz, 1H).

tert-Butyl 2-(chloromethyl)-1Н-1,3-benzodiazol-1-carboxylate (L)

A mixture of 2-(chloromethyl)-1H-1,3-benzodiazol (10 g, 0.06 mol), BOC2O (18 ml, 0.06 mol) and TEA (6.07 g, 0.06 mol) in DCM ( 304 ml) was cooled to 0°C. A catalytic amount of DMAP (0.73 g, 0.006 mol) was added and the reaction mixture was stirred at room temperature for 2 hours. The resulting mixture was diluted with EtOAc (150 ml), washed with saturated NaHCO ₃ solution (150 ml), (150 ml), dried (Na ₂ SO ₄ ), filtered and evaporated to give the crude product. Purification by flash column chromatography (eluting with a 5-10% EtOAc/heptane gradient) gave the title compound (7 g, 44%) as an off-white oil.

HPLC-MS (method H): [m/z]: 167.2 [M-Boc+H]+.

Diagram I-1 above:

ethyl 2-(2-{[(tert-butoxy)carbonyl]amino}ethyl)-4-methyl-1,3-thiazole-5-carboxylate (208)

Similar to general procedure 1, a suspension of tert-butyl (3-amino-3-thioxopropyl) carbamate (1.0 g, 5.34 mmol), CaCO ₃ (0.29 g, 2.93 mmol) and ethyl 2-chloro- 3-oxobutanoate (0.81 ml, 5.86 mmol) in EtOH (15 ml) was heated at 60° C. for 18 hours. The reaction mixture was cooled to room temperature and evaporated in vacuo. The resulting residue was partitioned between EtOAc and water and the phases were separated. The aqueous phase was extracted with ethyl acetate (2x80 ml) and the combined organic extracts washed with saturated sodium chloride solution (80 ml). The organic phase was dried (Na ₂ SO ₄ ), filtered and evaporated in vacuo. Purification by flash column chromatography (eluting with a 5-50% EtOAc/heptane gradient) gave the title compound (1.57 g, 94%) as a white solid.

1H NMR (CDCl ₃ , 500 MHz): δ [ppm] = 5.01 (brs, 1H), 4.34 (q, J=7.1 Hz, 2H), 3.57 (dt , J=5.5, 5.5 Hz, 2H), 3.15 (t, J=6.3 Hz, 2H), 2.73 (s, 3H), 1.47 (s, 9H), 1 .38 (t, J=7.1 Hz, 3H).

HPLC-MS (Method A): [m/z]: 315.10 [M+H]+.

Ethyl 2-(2-{[(tert-butoxy)carbonyl]amino}ethyl)-1,3-thiazole-5-carboxylate (209)

Similar to general procedure 1, tert-butyl K-(2-carbamothioylethyl)carbamate (1 g, 4.89 mmol), calcium carbonate (0.27 g, 3 mmol) and ethyl 2-chloro-3-oxopropanoate (0.81 g, 5 mmol) was combined in EtOH (15 ml) and the resulting mixture was heated at 60° C. for 18 h. More 2-chloro-3-oxopropanoate (0.81 g, 5 mmol) was added and the resulting mixture was heated at 80°C for another 5 hours the Reaction mixture was cooled to room temperature and evaporated in vacuum. The resulting residue was partitioned between water and EtOAc, and the resulting mixture was extracted with ethyl acetate (3x80 ml). The combined organic extracts were washed with saturated sodium chloride solution (80 ml), dried (Na ₂ SO ₄ ), filtered and evaporated in vacuo. Purification by flash column chromatography (gradient elution 20-79 039916

60% EtOAc/heptane) gave the crude title compound (743 mg) as a brown oil which was used in the next step without further purification.

HPLC-MS (method A): [m/z]: 301.05 [M+H] ⁺ .

2-(2-{[(tert-Butoxy)carbonyl]amino}ethyl)-4-methyl-1,3-thiazole-5-carboxylic acid (210)

Similar to general procedure 5, ethyl 2-(2-{[(tert-butoxy)carbonyl]amino}ethyl)-4-methyl-1,3thiazole-5-carboxylate (208) (1.57 g, 4.99 mmol) and LiOH (0.72 g, 30 mmol) in THF (30 ml) and water (15 ml) gave the title compound (1.23 g, 86%) as a white solid.

¹ H NMR (DMSO-d ₆ , 500 MHz): δ [ppm] = 6.99 (t, J=5.3 Hz, 1H), 3.28 (m, 2H), 3.04 ( t, J=6.7 Hz, 2H), 2.58 (s, 3H), 1.37 (s, 9H).

HPLC-MS (method A): [m/z]: 287.05 [M+H] ⁺ .

2-(2-{[(tert-Butoxy)carbonyl]amino}ethyl)-1,3-thiazole-5-carboxylic acid (211)

Similar to general procedure 5, crude ethyl 2-(2-{[(tert-butoxy)carbonyl]amino}ethyl)-1,3-thiazole5-carboxylate (209) (743 mg, 2.47 mmol) and LiOH (300 mg , 12 mmol) in THF (20 ml) and water (10 ml) gave the crude title compound (500 mg) as a brown oil which was used in the next step without further purification.

HPLC-MS (method A): [m/z]: 273.05 [M+H]+.

tert-Butyl yl)ethyl]carbamate (212)

N-[2-(5-{[(3-fluoropyridin-2-yl)methyl]carbamoyl}-4-methyl-1,3-thiazol-2-

Similar to general procedure 6, 2-(2-{[(tert-butoxy)carbonyl]amino}ethyl)-4-methyl-1,3-thiazole-5carboxylic acid (210) (0.457 g, 1.6 mmol), ( 3-fluoropyridin-2-yl)methanamine dihydrochloride (A2) (0.349 g, 1.76 mmol), DIPEA (0.92 ml, 5 mmol) and HATU (0.73 g, 2 mmol) in DCM (20 ml) gave the crude title compound (1.26 g) as a colorless oil.

HPLC-MS (method A): [m/z]: 395.1 [M+H] ⁺ .

tert-Butyl N-[2-(5-{[(3-fluoropyridin-2-yl)methyl]carbamoyl}-1,3-thiazol-2-yl)ethyl]carbamate (213)

As in general procedure 6, crude 2-(2-{[(tert-butoxy)carbonyl]amino}ethyl)-1,3-thiazole-5carboxylic acid (211) (500mg), (3-fluoropyridin-2-yl) methanamine dihydrochloride (A2) (402 mg, 2.02 mmol), DIPEA (1.06 ml, 6 mmol) and HATU (840 mg, 2 mmol) in DCM (30 ml) gave the crude title compound (953 mg, 87% pure) as a yellow residue after partial purification by flash column chromatography (gradient elution 20-100% EtOAc/heptane then 0-10% MeOH/EtOAc).

HPLC-MS (method A): [m/z]: 381.05 [M+H] ⁺ .

2-(2-Aminoethyl)-N-[(3-fluoropyridin-2-yl)methyl]-4-methyl-1,3-thiazole-5-carboxamide (214)

Similar to general procedure 2, crude tert-butyl N-[2-(5-{[(3-fluoropyridin-2yl)methyl]carbamoyl}-4-methyl-1,3-thiazol-2-yl)ethyl]carbamate (212 ) (1.26 g) and 12M HC (2 ml) in MeOH (20 ml) gave the title compound free base (471 mg) as a white solid after purification on an SCX-2 cartridge, washing with DCM and MeOH, followed by elution with 7n. a solution of ammonia in MeOH.

1H NMR (DMSO-d ₆ , 500 MHz): δ [ppm] = 8.56 (t, J=5.5 Hz, 1H), 8.39 (dt, J=4.6, 1, 3 Hz, 1H), 7.73-7.67 (m, 1H), 7.43-7.38 (m, 1H), 4.61-4.56 (m, 2H), 3.03 (t , J=7.0 Hz, 2H), 2.95 (t, J=6.5 Hz, 2H), 2.54 (s, 3H).

HPLC-MS (method A): [m/z]: 295.05 [M+H] ⁺ .

2-(2-Aminoethyl)-N-[(3-fluoropyridin-2-yl)methyl]-1,3-thiazole-5-carboxamide (215)

- 80 039916

Similar to general procedure 2, crude tert-butyl N-[2-(5-{[(3-fluoropyridin-2yl)methyl]carbamoyl}-1,3-thiazol-2-yl)ethyl]carbamate (213) (87% , 923 mg, 2.11 mmol) and 12M HC (2 ml) in

MeOH (20 ml) gave the title compound in free base form (389 mg) as a yellow residue after purification on an SCX-2 cartridge, washing with DCM and MeOH, followed by elution with 7N. a solution of ammonia in MeOH.

HPLC-MS (method A): [m/z]: 280.95 [M+H]+.

2-(2-{[2-(1H-1,3-Benzodiazol-2-yl)ethyl]amino}ethyl)-N-[(3-fluoropyridin-2-yl)methyl]-4-methyl-1, 3thiazole-5-carboxamide (illustrative example No. 7)

Similar to general procedure 8, 2-(2-aminoethyl)-N-[(3-fluoropyridin-2-yl)methyl]-4-methyl-1,3thiazole-5-carboxamide (214) (471 mg, 1.6 mmol ), N-(2-nitrophenyl)prop-2-enamide (D) (295 mg, 1.54 mmol) and DBU (0.26 mL, 2 mmol) gave a crude intermediate which was reacted with iron powder (280 mg , 5 mmol) in AcOH (5 mL) to give the title compound (115 mg, 20%) as a white solid after purification by basic prep-HPLC, followed by flash column chromatography (elution with a gradient of 0-30% MeOH/DXM).

¹ H NMR (Methanol-d4, 500 MHz): δ [ppm] = 8.38 (d, J=4.7 Hz, 1H), 7.61 (t, J=9.2 Hz, 1H ), 7.52-7.46 (m, 2H), 7.43-7.38 (m, 1H), 7.22-7.15 (m, 2H), 4.73 (s, 2H), 3.22-3.14 (m, 4H), 3.14-3.05 (m, 4H), 2.51 (s, 3H).

HPLC-MS (Method C): [m/z]: 439.1 [M+H] ⁺ .

2-(2-{[2-(1H-1,3-Benzodiazol-2-yl)ethyl]amino}ethyl)-N-[(3-fluoropyridin-2-yl)methyl]-1,3-thiazol- 5 carboxamide (illustrative example No. 8)

Similar to general procedure 8, 2-(2-aminoethyl)-N-[(3-fluoropyridin-2-yl)methyl]-1,3-thiazol-5carboxamide (215) (389 mg, 1.39 mmol), N- (2-nitrophenyl)prop-2-enamide (D) (267 mg, 1.39 mmol) and DBU (0.25 ml, 2 mmol) in MeCN (15 ml) gave a crude intermediate which was reacted with iron powder ( 230 mg, 4 mmol) in AcOH (5 mL) to give the title compound (49 mg, 11%) as a white solid after three basic prep-HPLC purifications.

1H NMR (Methanol-d ₄ , 500 MHz): δ [ppm] = 8.38 (dt, J=4.9, 1.3 Hz, 1H), 8.14 (s, 1H), 7 .65-7.59 (m, 1H), 7.52-7.47 (m, 2H), 7.44-7.39 (m, 1H), 7.21-7.17 (m, 2H) , 4.75 (d, J=1.6 Hz, 2H), 3.23 (t, J=6.8 Hz, 2H), 3.153.07 (m, 6H).

HPLC-MS (method C): [m/z]: 425.1 [M+H] ⁺ .

The above general scheme 24:

methyl 3-amino-2-hydroxypropanoate hydrochloride (332) o

MeO^Y^NH^ HCl he

Thionyl chloride (1.8 ml, 20 mmol) was added dropwise to ice-cold MeOH (60 ml) and stirred for 5 minutes. 3-Amino-2-hydroxypropanoic acid (1.04 g, 9.9 mmol) was added and the reaction mixture was stirred at room temperature for 18 h. The reaction mixture was evaporated in vacuo to give the title compound (1.54 g, quant.) as a yellow oil.

1H NMR (DMSO-66, 250 MHz): δ [ppm] = 8.12 (s, 3H), 6.36 (s, 1H), 4.39 (dt, J=8.8, 4 .6 Hz, 1H), 3.69 (s, 3H), 3.24-3.05 (m, 1H), 3.00-2.84 (m, 1H).

Methyl 3-(3-{[(tert-butoxy)carbonyl]amino}propanamido)-2-hydroxypropanoate (333) o o

Similar to general procedure 13, methyl 3-amino-2-hydroxypropanoate hydrochloride (332) (4.35 g, 27.96 mmol), 3-{[(t-butoxy)carbonyl]amino}propanoic acid (5.82 g, 30.76 mmol), TEA (4.68 ml, 34 mmol) and DCC (5.77 g, 28 mmol) in DCM (80 ml) gave the title compound (5.12 g, 63%) as a pale -yellow oil after purification by flash column chromatography (eluting with a 0-100% EtOAc/heptane gradient).

- 81 039916

1H NMR (CDCl ₃ , 500 MHz): δ [ppm] = 6.06 (s, 1H), 5.18 (s, 1H), 4.32 (q, J=4.9 Hz, 1H ), 3.83 (s, 3H),

3.72-3.60 (m, 2H), 3.46 (d, J=5.4 Hz, 1H), 3.45-3.38 (m, 2H), 2.47-2.36 ( m, 2H), 1.46 (s, 9H).

HPLC-MS (Method A): [m/z]: 313.00 [M+Na]+.

Methyl 2-(2-{[(tert-butoxy)carbonyl]amino}ethyl)-4,5-dihydro-1,3-oxazole-5-carboxylate (334)

Similar to general procedure 14, methyl 3-(3-{[(tert-butoxy)carbonyl]amino}propanamido)-2hydroxypropanoate (333) (3.63 g, 12.5 mmol), DAST (1.98 ml, 15 mmol ) and K ₂ CO ₃ (3.46 g, 25 mmol) in DCM (100 ml) gave the title compound (3.4 g, 99%) as a colorless oil after purification by flash column chromatography (gradient elution 40-100% EtOAc/heptane).

1H NMR (CDCl3, 250 MHz): δ [ppm] = 4.96 (dd, J=10.8, 6.5 Hz, 1H), 4.24-4.09 (m, 1H), 4.04-3.88 (m, 1H), 3.83 (s, 3H), 3.58-3.38 (m, 2H), 2.57 (d, J=4.8 Hz, 2H) , 1.47 (s, 9H).

HPLC-MS (method M): [m/z]: 272.95 [M+H] ⁺ .

General Method 16:

methyl 2-(2-{[(tert-butoxy)carbonyl]amino}ethyl)-1,3-oxazole-5-carboxylate (335)

Methyl 2-(2-{[(tert-butoxy)carbonyl]amino}ethyl)-4,5-dihydro-1,3-oxazole-5-carboxylate solution (334) (3.2 g, 11.75 mmol) , NBS (2.3 g, 12.93 mmol) and AIBN (0.19 g, 1.18 mmol) in DCE (30 ml) were heated at 80°C for 1.5 h. The reaction mixture was cooled to room temperature , extinguished with saturated aqueous NaHCO ₃ and was extracted with DCM (3x80 ml). The combined organic extracts were dried (Na ₂ SO ₄ ), filtered and evaporated in vacuo. Purification by flash column chromatography (eluting with a 0-80% EtOAc/heptane gradient) gave the crude title compound (1.25 g, 29%, 75% pure) as an orange residue. The compound was used in the next step without further purification.

1H NMR (CDCl3, 250 MHz): δ [ppm] = 7.69 (s, 1H), 5.17-4.98 (m, 1H), 3.94 (s, 3H), 3, 62 (q, J=6.2 Hz, 2H), 3.05 (t, J=6.2 Hz, 2H), 1.46 (s, 9H).

HPLC-MS (method M): [m/z]: 271.00 [M+H] ⁺ .

2-(2-{[(tert-butoxy)carbonyl]amino}ethyl)-1,3-oxazole-5-carboxylic acid (336)

As in general procedure 5, crude methyl 2-(2-{[(tert-butoxy)carbonyl]amino}ethyl)-1,3oxazole-5-carboxylate (335) (1.24 g, 3.44 mmol, 75% pure ) and LiOH (0.329 mg, 13.76 mmol) in THF (20 mL) and water (20 mL) gave the title compound (0.51 g, 44%) as a light orange residue. The compound was used in the next step without further purification.

1H NMR (CDCl3, 250 MHz): δ [ppm] = 7.76 (s, 1H), 5.14 (s, 1H), 3.71-3.56 (m, 2H), 3, 09 (t, J=6.2 Hz, 2H), 1.46 (s, 9H).

HPLC-MS (method M): [m/z]: 256.95 [M+H] ⁺ .

tert-Butyl N-[2-(5-{[(3-fluoropyridin-2-yl)methyl]carbamoyl}-1,3-oxazol-2-yl)ethyl]carbamate (337)

Similar to general procedure 6, 2-(2-{[(tert-butoxy)carbonyl]amino}ethyl)-1,3-oxazole-5carboxylic acid (336) (513 mg, 2 mmol), (3-fluoropyridine-2- yl)methanamine dihydrochloride (A2) (479 mg, 2.4 mmol), DIPEA (1.15 ml, 6.61 mmol) and HATU (837 mg, 2.2 mmol) in DCM (20 ml) gave the crude indicated in title compound (1.08 g, 74%, 50% pure) as a yellow residue. The compound was used in the next step without purification.

HPLC-MS (method M): [m/z]: 365.05 [M+H] ⁺ .

2-(2-Aminoethyl)-N-[(3-fluoropyridin-2-yl)methyl]-1,3-oxazole-5-carboxamide (338)

Similar to general procedure 2, crude tert-butyl N-[2-(5-{[(3-fluoropyridin-2yl)methyl]carbamoyl}-1,3-oxazol-2-yl)ethyl]carbamate (337) (1, 08 g, 1.48 mmol) and 12M HC (1 ml) in MeOH (10 ml) gave the title compound in free base form (444 mg, 57%) as a pale yellow solid after conversion to free base form using SCX-2 cartridge (10 g), washing with DCM and MeOH, followed by elution with 7N. ammonia solution in

Meon.

¹ H NMR (DMSO-d ₆ , 250 MHz): δ [ppm] = 8.99 (t, J=5.7 Hz, 1H), 8.39 (dt, J=4.6, 1 .5 Hz, 1H), 7.78-7.64 (m, 2H), 7.48-7.36 (m, 1H), 4.61 (dd, J=5.8, 1.6 Hz, 2H), 3.07-2.86 (m, 4H).

HPLC-MS (method M): [m/z]: 264.95 [M+H]+.

2-(2-{[2-(1H-1,3-benzodiazol-2-yl)ethyl]αmino}ethyl)-N-[(3-fluoropyridin-2-yl)methyl]-1,3-oxazol- 5carboxamide illustrative example no. 1)

Similar to general procedure 8, 2-(2-aminoethyl)-N-[(3-fluoropyridin-2-yl)methyl]-1,3-oxazole-5carboxamide (338) (444 mg, 1.68 mmol), N- (2-nitrophenyl)prop-2-enamide (D) (323 mg, 1.68 mmol) and DBU (301 μl, 2.02 mmol) in MeCN (10 ml) gave a crude intermediate which was reacted with iron powder ( 343 mg, 6.13 mmol) and AcOH (10 mL) to give the title compound as a pale yellow solid (166 mg, 25%) after purification by flash column chromatography (eluting with a gradient of 0-40% MeOH/DCM ), followed by a basic prep HPLC.

¹ H NMR (DMSO-d ₆ , 500 MHz): δ [ppm] = 12.12 (s, 1H), 8.96 (t, J=5.8 Hz, 1H), 8.38 ( dt, J=4.6, 1.4 Hz, 1H), 7.73-7.66 (m, 2H), 7.53-7.37 (m, 3H), 7.11 (dt, J= 6.0, 3.5 Hz, 2H), 4.61 (dd, J=5.7, 1.4 Hz, 2H), 3.04-2.89 (m, 8H).

HPLC-MS (method B): [m/z]: 409.1 [M+H] ⁺ .

2-{2-[(1H-1,3-Benzodiαzol-2-ylmethyl)α-mino]ethyl}-N-[(3-fluoro-pyridin-2-yl)methyl]-1,3-oxazol-5-carboxamide (Illustrative Example No. four)

Similar to general procedure 3, 2-(2-Aminoethyl)-N-[(3-fluoropyridin-2-yl)methyl]-1,3-oxazole-5-carboxamide (338) (250 mg, 0.83 mmol, 88% pure ), 1H-benzimidazole-2-carbaldehyde (170 mg, 1.17 mmol), DIPEA (0.44 ml, 2.5 mmol) and MgSO ₄ (150 mg, 1.25 mmol) in MeOH (10 ml) at room temperature for 16 h gave an intermediate which was further reacted with NaBH ₄ (47 mg, 1.25 mmol) to give the title compound (156 mg, 48%) as a white solid after purification by column flash. -chromatography [eluting with a gradient of 0-5% (7N NH ₃ in MeOH)/DCM], followed by a basic prep-HPLC.

¹ H NMR (DMSO-d ₆ , 500 MHz): δ [ppm] = 12.17 (brs, 1H), 8.95 (t, J=5.8 Hz, 1H), 8, 39-8.34 (m, 1H), 7.72-7.65 (m, 2H), 7.54-7.46 (m, 1H), 7.46-7.36 (m, 2H), 7.17-7.06 (m, 2H), 4.63-4.56 (m, 2H), 3.93 (s, 2n), 3.02-2.94 (m, 4H), 2, 57 (broad s, 1H).

HPLC-MS (Method D): [m/z]: 395.2 [M+H] ⁺ .

The above general scheme 27:

tert-butyl 3-[(2-hydroxy-3-methoxy-3-oxopropyl)carbamoyl]azetidine-1-carboxylate (354)

Similar to general procedure 13, methyl 3-amino-2-hydroxypropanoate hydrochloride (2.9 g, 18.64 mmol), 1-(tert-butoxycarbonyl)azetidine-3-carboxylic acid (4.13 g, 20.5 mmol) , TEA (5.45 ml, 39.14 mmol) and DCC (4.04 g, 19.57 mmol) in DCM (80 ml) gave the title compound (4.47 g, 79%) as a viscous light -yellow oil after purification by flash column chromatography (gradient elution 0-100% EtOAc/heptane followed by 0-3% MeOH/EtOAc).

¹ H NMR (CDCl ₃ , 500 MHz): δ [ppm] = 5.92 (s, 1H), 4.32 (dd, J=5.3, 4.3 Hz, 1H), 4, 14-4.03 (m, 5H), 3.84 (s, 3H), 3.74-3.63 (m, 2H), 3.24-3.14 (m, 1H), 1.46 ( s, 9H).

HPLC-MS (method M): [m/z]: 325.00 [M+Na] ⁺ .

Methyl 2-{1-[(tert-butoxy)carbonyl]azetidin-3-yl}-4,5-dihydro-1,3-oxazole-5-carboxylate (355)

Similar to general procedure 14, t-butyl 3-[(2-hydroxy-3-methoxy-3oxopropyl)carbamoyl]azetidine-1-carboxylate (354) (4.47 g, 14.79 mmol) and DAST (2.15 ml , 16.26

- 83039916 mmol) in DCM (100 mL) followed by quenching with K2CO ₃ (4.09 g, 29.57 mmol) gave the title compound (1.61 g, 38%) as a pale yellow oil after purification by flash column chromatography (elution gradient 20-100% EtOAc/heptane, then 1%

MeOH/EtOAc).

¹ H NMR (CDCl3, 250 MHz): δ [ppm] = 5.00 (dd, J=10.8, 6.7 Hz, 1H), 4.27-4.08 (m, 5H) , 4.00 (ddd, J=14.7, 6.7, 1.0 Hz, 1H), 3.83 (s, 3H), 3.55-3.39 (m, 1H), 1.46 (s, 9H).

Methyl 2-{1-[(tert-butoxy)carbonyl]azetidin-3-yl}-1,3-oxazole-5-carboxylate (356) o

MeO^X^o /—\J\IBoc N N'

Similar to general procedure 16, methyl 2-{1-[(tert-butoxy)carbonyl]azetidin-3-yl}-4,5-dihydro1,3-oxazole-5-carboxylate (355) (1.61 g, 5. 66 mmol), NBS (1.11 g, 6.23 mmol) and AIBN (0.11 g, 0.57 mmol) in DCE (60 ml) at 80° C. for 1.5 h gave the title compound (756 mg, 45%) as a pale yellow oil after purification by flash column chromatography (eluting with a 0-80% EtOAc/heptane gradient).

Ή NMR (CDCl ₃ , 250 MHz): δ [ppm] = 7.74 (s, 1H), 4.38-4.23 (m, 4H), 4.03-3.83 (m, 5H), 1.48 (s, 9H).

HPLC-MS (method M): [m/z]: 304.95 [M+Na]+.

2-{1-[(tert-Butoxy)carbonyl]azetidin-3-yl}-1,3-oxazole-5-carboxylic acid (3 57) o ηοΆ^ο / CjMBoc Ν Ν'

Similar to general procedure 5, methyl 2-{1-[(tert-butoxy)carbonyl]azetidin-3-yl}-1,3-oxazol-5carboxylate (356) (750 mg, 2.66 mmol) and LiOH (382 mg , 15.94 mmol) in THF/water (20 ml/20 ml) gave the title compound (710 mg, 99%) as a pale yellow oil. The resulting title compound was used in the next step without purification.

Ή NMR (CDCl ₃ , 250 MHz): δ [ppm] = 7.83 (s, 1H), 4.40-4.25 (m, 4H), 4.04-3.94 (m, 1H), 1.48 (s, 9H).

HPLC-MS (method M): [m/z]: 290.90 [M+Na]+.

tert-Butyl 3-(5-{[(3-fluoropyridin-2-yl)methyl]carbamoyl}-1,3-oxazol-2-yl)azetidine-1 carboxylate(358)

Similar to general procedure 6, 2-{1-[(tert-butoxy)carbonyl]azetidin-3-yl}-1,3-oxazole-5carboxylic acid (357) (0.74 g, 2.77 mmol), (3 -fluoropyridin-2-yl)methanamine dihydrochloride (A2) (0.61 g, 3.04 mmol), DIPEA (1.59 ml, 9.13 mmol) and HATU (1.16 g, 3.04 mmol) in DCM (30 mL) gave the title compound (2.26 g, 45% pure) as a pale yellow oil after purification by flash column chromatography (gradient elution 20-100% EtOAc/heptane, then 2-10% MeOH/EtOAc). The resulting material was used in the next step without further purification.

¹ H NMR (CDCl ₃ , 250 MHz): δ [ppm] = 8.44 (dt, J=4.8, 1.3 Hz, 1H), 7.72 (s, 1H), 7, 60 (s, 1H), 7.53-7.42 (m, 1H), 7.38-7.31 (m, 1H), 4.84 (dd, J=4.8, 1.5 Hz, 2H), 4.42-4.24 (m, 4H), 4.06-3.91 (m, 1H), 1.49 (s, 9H).

HPLC-MS (method M): [m/z]: 377.05 [M+H] ⁺ .

2-(Azetidin-3-yl)-N-[(3-fluoropyridin-2-yl)methyl]-1,3-oxazol-5-carboxamide (3 5 9)

F

Similar to general procedure 2, tert-butyl 3-(5-{[(3-fluoropyridin-2-yl)methyl]carbamoyl}-1,3oxazol-2-yl)azetidine-1-carboxylate (358) (45%, 2 .26 g, 2.7 mmol) and 12M HC (2 ml) in MeOH (20 ml) gave the title compound (644 mg, 86%) as a white solid after conversion to the free base form with SCX-2 cartridge (10 g), washing with DCM and MeOH, followed by elution with 7N. a solution of ammonia in MeOH.

¹ H NMR (DMSO-d ₆ , 500 MHz): δ [ppm] = 9.03 (t, J=5.6 Hz, 1H), 8.39 (dt, J=4.5, 1 .3 Hz, 1H), 7.74 (s, 1H), 7.70 (ddd, J=10.0, 8.4, 1.2 Hz, 1H), 7.41 (dt, J=8, 6, 4.4 Hz, 1H), 4.63-4.60 (m, 2H), 4.11-4.02 (m, 1H), 3.85 (t, J=7.6 Hz, 2H ), 3.78 (t, J=8.2 Hz, 2H).

HPLC-MS (method M): [m/z]: 276.95 [M+H] ⁺ .

2-{1-[2-(1H-1,3-Benzodiazol-2-yl)ethyl]azetidin-3-yl}-N-[(3-fluoropyridin-2-yl)methyl]-1,3-oxazol5 -carboxamide (illustrative example No. 2)

- 84 039916

Similar to general procedure 8, 2-(azetidin-3-yl)-N-[(3-fluoropyridin-2-yl)methyl]-1,3-oxazole-5-carboxamide (359) (255 mg, 0.92 mmol), N-(2-nitrophenyl)prop-2-enamide (D) (186 mg, 0.97 mmol) and DBU (145 μl, 0.97 mmol) in MeCN (10 ml) gave a crude intermediate which reacted with the powder iron (167 mg, 3 mmol) in AcOH (4 ml) to give the title compound (140 mg, 44%) as a white solid after purification by basic prep-HPLC followed by flash column chromatography (gradient elution 0-40% MeOH/DCM).

¹ H NMR (MeOD, 500 MHz): δ [ppm] = 8.36 (dt, J=4.7, 1.2 Hz, 1H), 7.69 (s, 1H), 7.62 (ddd, J=9.8, 8.4, 1.2 Hz, 1H), 7.52 (s, 2H), 7.41 (dt, J=8.6, 4.4 Hz, 1H), 7.24-7.17 (m, 2H), 4.77 (d, J=1.6 Hz, 2H), 3.96-3.87 (m, 1H), 3.76 (t, J= 7.9 Hz, 2H), 3.58 (t, J=7.3 Hz, 2H), 3.08-3.02 (m, 2H), 3.01-2.95 (m, 2H).

HPLC-MS (method B): [m/z]: 421.1 [M+H]+.

2-{1-[2-(4-Fluoro-1H-1,3-benzodiazol-2-yl)ethyl]azetidin-3-yl}-N-[(3-fluoropyridin-2-yl)methyl]-1 ,3oxazole-5-carboxamide (illustrative example No. 3)

Similar to general procedure 8, 2-(azetidin-3-yl)-N-[(3-fluoropyridin-2-yl)methyl]-1,3-oxazole-5-carboxamide (359) (400 mg, 1.45 mmol), N-(3-fluoro-2-nitrophenyl)prop-2-enamide (G) (335 mg, 1.59 mmol) and DBU (238 μl, 1.59 mmol) in MeCN (10 ml) gave the crude intermediate, which was partially purified by flash column chromatography (eluting with a 0-20% MeOH/DCM gradient). The resulting intermediate was further reacted with iron powder (76 mg) in AcOH (3 mL) to give the title compound (52 mg, 34%) as a beige solid after purification by flash column chromatography (eluting with a 0-35% gradient). MeOH/DXM).

¹ H NMR (MeOD, 500 MHz): δ [ppm] = 8.38-8.34 (m, 1H), 7.69 (s, 1H), 7.65-7.60 (m, 1H), 7.44-7.39 (m, 1H), 7.31 (d, J=8.2 Hz, 1H), 7.21-7.15 (m, 1H), 6.98-6 .92 (m, 1H), 4.77 (d, J=1.6 Hz, 2H), 3.92 (n, J=7.1 Hz, 1H), 3.77 (t, J=8, 0 Hz, 2H), 3.58 (t, J=7.4 Hz, 2H), 3.08-3.02 (m, 2H), 3.02-2.96 (m, 2H).

HPLC-MS (method d): [m/z]: 439.2 [M+H] ⁺ .

2-{1-[2-(5-Fluoro-1H-1,3-benzodiazol-2-yl)ethyl]azetidin-3-yl}-N-[(3-fluoropyridin-2-yl)methyl]-1 ,3oxazole-5-carboxamide (illustrative example No. 5)

Similar to general procedure 8, 2-(azetidin-3-yl)-N-[(3-fluoropyridin-2-yl)methyl]-1,3-Oksazol-5carboxamide (359) (581 mg, 2.1 mmol), N-(5-fluoro-2-nitrophenyl)prop-2-enamide (J) (371 mg, 1.77 mmol) and DBU (346 μl, 2.31 mmol) in MeCN (30 ml) gave the crude intermediate, which was reacted with iron powder (186 mg, 3.34 mmol) in AcOH (20 mL) to give the title compound (205 mg, 56%) as a white solid after purification by basic prep-HPLC.

¹ H NMR (MeOD, 500 MHz): δ [ppm] = 8.37 (dt, J=4.6, 1.2 Hz, 1H), 7.69 (s, 1H), 7.62 (ddd, J=9.8, 8.4, 1.2 Hz, 1H), 7.51-7.45 (m, 1H), 7.41 (dt, J=8.7, 4.4 Hz , 1H), 7.22 (dd, J=9.1, 2.1 Hz, 1H), 7.03-6.96 (m, 1H), 4.77 (d, J=1.6 Hz, 2H), 3.95-3.86 (m, 1H), 3.76 (t, J=8.0 Hz, 2H), 3.57 (t, J=7.4 Hz, 2H), 3, 06-3.01 (m, 2H), 3.00-2.94 (m, 2H).

HPLC-MS (method B): [m/z]: 439.2 [M+H] ⁺ .

The above general scheme 1:

general technique 1:

ethyl 2-(2-{[(tert-butoxy)carbonyl]amino}ethyl)-1,3-thiazole-4-carboxylate (1)

To a suspension of ethyl 3-bromo-2-oxopropanoate (12.35 mL, 107.69 mmol) and tert-butyl (3-amino-3-thioxopropyl)carbamate (20 g, 97.9 mmol) in EtOH (200 mL) was added portionwise CaCO ₃ (5.3 g, 52.87 mmol) and the reaction mixture was stirred at room temperature for 12 h. ₃ (100 ml). The organic layer was separated and washed with water (100 ml), brine (100 ml), dried (MgSO ₄ ), filtered and evaporated in vacuo to give the desired product. Purification by flash column chromatography (isocratic elution with 20%

EtOAc/heptane) gave the title compound (22 g, 69.6%) as a yellow solid.

¹ H NMR (Methanol-b ₄ , 250 MHz): δ [ppm] = 8.29 (s, 1H), 4.39 (q, J=7.1 Hz, 2H), 3.47 ( t, J=6.5 Hz, 2H), 3.22 (t, J=6.5 Hz, 2H), 1.41 (d, J=6.2 Hz, 14H).

HPLC-MS (method A): [m/z]: 301.0 [M+H] ⁺ .

Methyl 2-(2-{[(tert-butoxy)carbonyl]amino}ethyl)-1,3-thiazole-4-carboxylate (2)

Similar to general procedure 1, tert-Butyl (3-amino-3-thioxopropyl) carbamate (10 g, 48.95 mmol), methyl 3-bromo-2-oxopropanoate (5.73 ml, 53.85 mmol) and CaCO ₃ (0.9 ml, 26.43 mmol) in EtOH (120 ml) gave the title compound (10.2 g, 60%, 83% pure) as a yellow solid after purification by flash column chromatography (elution with gradient 20-80% EtOAc/heptane).

HPLC-MS (Method A): [m/z]: 286.9 [M+H]+.

Methyl 2-(2-{[(tert-butoxy)carbonyl]amino}ethyl)-5-methyl-1,3-thiazole-4-carboxylate (3)

Similar to general procedure 1, tert-butyl N-(2-carbamothioylethyl)carbamate (0.89 g, 4.35 mmol), methyl 3-bromo-2-oxobutanoate (0.93 g, 4.78 mmol) and CaCO ₃ (0.23 g, 2 mmol) in EtOH (15 mL) gave the title compound (0.769 g, 58%) as a yellow oil after purification by flash column chromatography (eluting with a 10-60% EtOAc/heptane gradient) .

1H NMR (CDCl3, 250 MHz): δ [ppm] = 4.88 (s, 1H), 3.95 (s, 3H), 3.55 (q, J=6.5 Hz, 2H) , 3.17 (t, J=6.5 Hz, 2H), 2.76 (s, 3H), 1.46 (s, 9H).

HPLC-MS (method A): [m/z]: 301.05 [M+H] ⁺ .

Ethyl 2-(3-{[(tert-butoxy)carbonyl]amino}propyl)-1,3-thiazole-4-carboxylate (4)

Similar to general procedure 1, tert-butyl N-(3-carbamothioylpropyl)carbamate (535 mg, 2.45 mmol), ethyl 3-bromo-2-oxopropanoate (0.31 ml, 2.7 mmol) and CaCO ₃ (132 mg, 1.32 mmol) in EtOH (10 mL) gave the title compound (726 mg, 93%) as a yellow oil after purification by flash column chromatography (eluting with a 0-50% EtOAc/heptane gradient).

1H NMR (DMSO-a ₆ , 500 MHz): δ [ppm] = 8.38 (s, 1H), 6.90 (s, 1H), 4.29 (q, J=7.1 Hz , 2H), 3.15-2.90 (m, 4H), 1.83 (m, 2H), 1.38 (s, 9H), 1.30 (t, J=7.1 Hz, 3H) .

HPLC-MS (method A): [m/z]: 315 [M+H] ⁺ .

General procedure 2: methyl 2-(2-aminoethyl)-1,3-thiazole-4-carboxylate (5)

A 4M solution of HC in dioxane (44 ml, 176 mmol) was added to a solution of methyl 2-(2-{[(tert-butoxy)carbonyl]amino}ethyl)-1,3-thiazole-4-carboxylate (2) (10.2 g , 35.62 mmol) in dioxane, and the resulting mixture was stirred at room temperature for 12 h, then at 40° C. for 24 h. The resulting mixture was cooled to room temperature and evaporated in vacuo. The residue was dissolved in DCM (20 ml) and washed with saturated NaHCO ₃ solution (3x10 ml). The combined aqueous phases were re-extracted with diethyl ether (3x100 ml), the combined organic phases were dried (MgSO ₄ ), filtered and evaporated in vacuo to give the title compound (1.96 g, 30%) as a brown solid.

HPLC-MS (method A): [m/z]: 186.9 [M+H] ⁺ .

General technique carboxylate (6)

3: methyl 2-{2-[(1H-1,3-benzodiazol-2-ylmethyl)amino]ethyl}-1,3-thiazol-4-

methyl suspension

2-(2-aminoethyl)-1,3-thiazole-4-carboxylate (5) (1.96 g, 10.52 mmol), 1H-86 039916 benzimidazole-2-carbaldehyde (2.31 g, 15.79 mmol) and DIPEA (1.83 ml, 10.52 mmol) in MeOH (100 ml) were stirred at room temperature for 12 h. The reaction mixture was cooled to 0°C, NaBH ₄ (0.597 g, 15.79 mmol) was added , and the resulting mixture was stirred at room temperature for 2 hours, the Reaction mixture was evaporated in vacuum, the resulting residue was dissolved in EtOAc (100 ml) and washed with saturated solution of Na ₂ CO ₃ (2x50 ml). The combined aqueous layers were extracted with ethyl acetate (3x50 ml) and the combined organic layers were dried (MgSO ₄ ), filtered and evaporated in vacuo. Purification by flash column chromatography (KP-NH, eluting with a 0-10% MeOH/DCM gradient) gave the title compound (1.4 g, 38%, 90% pure) as a brownish solid.

1H NMR (Methanol-d ₄ , 250 MHz): δ [ppm] = 8.27 (s, 1H), 7.60-7.49 (m, 2H), 7.29-7.17 ( m, 2H), 4.09 (s, 2H), 3.92 (s, 3H), 3.26 (t, J=6.3 Hz, 2H), 3.10 (t, J=6.8 Hz, 2H).

HPLC-MS (method A): [m/z]: 317 [M+H]+.

General Procedure 4: tert-butyl 2-({[(tert-butoxy)carbonyl]({2-[4-(methoxycarbonyl)-1,3-thiazol-2yl]ethyl})amino}methyl)-1H-1, 3-benzodiazol-1-carboxylate (7)

In a solution of methyl 2-{2-[(1H-1,3-benzodiazol-2-ylmethyl)amino]ethyl}-1,3-thiazole-4-carboxylate (6) (74%, 2.94 g, 6, 88 mmol), Boc ₂ O (3.75 g, 17.19 mmol) and TEA (2.38 ml, 17.19 mmol) in THF (60 ml) were added DMAP (168 mg, 1.38 mmol), and the reaction mixture was stirred at room temperature for 16 hours. The reaction mixture was evaporated to dryness, diluted with EtOAc (100 ml) and washed with water (3x50 ml). The organic layer was dried over MgSO ₄ , filtered and evaporated to dryness. The resulting crude residue was purified by flash column chromatography eluting with 0-100% EtOAc in heptane to give 3.8 g of the desired product.

General Procedure 5: 2-(2-{[(tert-butoxy)carbonyl]({1-[(tert-butoxy)carbonyl]-1H-1,3benzodiazol-2-yl}methyl)amino}ethyl)-1, 3-thiazole-4-carboxylic acid (8) o

O ^ OS (CH ₃ ) s

Η

Lithium hydroxide (0.48 mg, 20.08 mmol) was added to a solution of tert-butyl 2-({[(tert-butoxy)carbonyl]({2-[4-(methoxycarbonyl)-1,3-thiazol-2-yl] ethyl})amino}methyl)-1H-1,3-benzodiazol-1carboxylate (7) (3.8 g, 6.69 mmol) in THF/water (40 ml/10 ml) at 0°C. The reaction mixture was stirred at room temperature for 48 hours. The resulting mixture was evaporated in vacuo and acidified to pH ~3-4 with AcOH. The reaction mixture was extracted with a mixture of THF/EtOAc (3:1, 3x50 ml). The combined organic extracts were washed with saturated sodium chloride solution (100 ml), dried (MgSO ₄ ), filtered, evaporated in vacuo and azeotropically dried with heptane (3x50 ml) to give the title compound (2.4 g, 84.6%) in yellow foam.

1H NMR (Methanol-d4, 250 MHz): δ [ppm] = 8.15 (d, J=17.0 Hz, 1H), 7.69 (s, 2H), 7.27 (dd, J=6.1, 3.2 Hz, 2H), 4.79 (s, 2H), 3.87-3.74 (m, 2H), 3.40-3.33 (m, 3H), 1 .38-1.01 (m, 10H).

HPLC-MS (method A): [m/z]: 403 [M+H] ⁺ .

General Procedure 6: tert-Butyl N-(1H-1,3-benzodiazol-2-ylmethyl)-N-(2-{4-[(pyridin-2ylmethyl)carbamoyl]-1,3-thiazol-2-yl} ethyl)carbamate (9)

To a stirred solution of 2-{2-[(1H-1,3-benzodiazol-2-ylmethyl)[(tert-butoxy)carbonyl]amino]ethyl}-1,3-thiazole-4-carboxylic acid (8) (3 g, 7.08 mmol), 1-(pyridin-2yl)methanamine (1.1 ml, 10.62 mmol), DIPEA (3.7 ml, 21.24 mmol) and DMF (50 ml) HATU was added at room temperature ( 5.39 g, 14.16 mmol). The reaction mixture was allowed to stir at room temperature for 16 hours. The mixture was diluted with EtOAc (100 ml), washed with saturated aqueous NaHCO ₃ (3x50 ml) and saturated sodium chloride solution (3x50 ml). The organic layer was separated, dried (MgSO ₄ ), filtered and evaporated to dryness. The resulting crude residue was purified by flash column chromatography (kp-NH, eluting with a gradient of 20-100%

- 87 039916

EtOAc in heptane) and then azeotropically dried with heptane to give the title compound (2.2 g, 62%) as a yellow foam.

1H NMR (MeOD, 500 MHz): δ [ppm] = 8.49 (d, J=4.4 Hz, 1H), 8.10 (s, 1H), 7.80 (td, J= 7.8, 1.7Hz, 1H),

7.54 (s, 2H), 7.42 (d, J=7.9 Hz, 1H), 7.31 (dd, J=7.1, 5.2 Hz, 1H), 7.26-7 .20 (m, 2H), 4.75 (d, J=12.3 Hz, 2H),

4.70 (s, 2H), 3.92-3.79 (m, 2H), 3.36 (d, J=8.1 Hz, 1H), 1.43-1.25 (m, 10H) .

HPLC-MS (Method D): [m/z]: 493.1 [M+H]+.

tert-Butyl N-(1H-1,3-benzodiazol-2-ylmethyl)-N-(2-{4-[(cyclohexylmethyl)carbamoyl]-1,3-thiazol2-yl}ethyl)carbamate (10)

n

Similar to general procedure 6, 2-{2-[(1H-1,3-benzodiazol-2-ylmethyl)[(tert-butoxy)carbonyl]amino]ethyl}-1,3-thiazole-4-carboxylic acid (8) (99 .8 mg, 0.248 mmol), cyclohexylmethanamine (33.69 mg, 0.298 mmol), DIPEA (96.16 mg, 0.744 mmol) and HATU (113.16 mg, 0.298 mmol) in DMF (4 ml) at room temperature in gave the title compound (116 mg, 47% pure) as an off-white oil over 1 hour after purification by flash column chromatography (eluting with a 0-10% MeOH/DCM gradient). The resulting title compound was used in the next step without further purification.

HPLC-MS (method H): [m/z]: 498.7 [M+H] ⁺ .

tert-Butyl N-(1H-1,3-benzodiazol-2-ulmethyl)-N-{2-[4-(1,2,3,4-tetrahydroisoquinoline-2-carbonyl)1,3-thiazole-2- yl]ethyl}carbamate (11)

Similar to general procedure 6, 2-{2-[(1H-1,3-benzodiazol-2-ylmethyl)[(tert-butoxy)carbonyl]amino]ethyl}-1,3-thiazole-4-carboxylic acid (8) (99 .8 mg, 0.248 mmol), 1,2,3,4tetrahydroisoquinoline (39.64 mg, 0.298 mmol), DIPEA (96.16 mg, 0.744 mmol) and HATU (113.16 mg, 0.298 mmol) in DMF (4 ml) at room temperature for 1 hour gave the title compound (124 mg, 59% pure) as an off-white oil after purification by flash column chromatography (eluting with a 0-10% MeOH/DCM gradient). The resulting title compound was used in the next step without further purification.

HPLC-MS (method H): [m/z]: 518.7 [M+H] ⁺ .

tert-Butyl N-(1H-1,3-benzodiazol-2-ylmethyl)-N-(2-{4-[(thiophen-2-ylmethyl)carbamoyl]-1,3-thiazol2-yl}ethyl)carbamate ( 12)

To a solution of 2-{2-[(1H-1,3-benzodiazol-2-ylmethyl)[(tert-butoxy)carbonyl]amino]ethyl}-1,3-thiazole-4carboxylic acid (8) (150 mg, 0.373 mmol) in DMF (10 ml) was added 1H-1,2,3-benzotriazol-1-ol (50 mg, 0.373 mmol) and EDC:HCl (71 mg, 0.373 mmol) at 0°C. The reaction mixture was allowed to stir for 15 minutes, then TEA (38 mg, 0.373 mmol) was added followed by thiophen-2-ylmethanamine (42 mg, 0.373 mmol). The reaction mixture was allowed to warm to room temperature and stirred overnight. The resulting title compound (185 mg, 16% pure) was obtained after work-up according to General Method 6. It was used in the next step without purification.

HPLC-MS (method H): [m/z]: 498.6 [M+H] ⁺ .

tert-Butyl N-(1H-1,3-benzodiazol-2-ul-methyl)-N-(2-{4-[benzyl(methyl)carbamoyl]-1,3-thiazol-2yl}ethyl)carbamate (13 )

n

Similar to general procedure 6, 2-{2-[(1H-1,3-benzodiazol-2-ylmethyl)[(tert-butoxy)carbonyl]amino]ethyl}-1,3-thiazole-4-carboxylic acid (8) (99 .8 mg, 0.248 mmol), benzyl(methyl)amine (36.06 mg, 0.298 mmol), DIPEA (96.16 mg, 0.744 mmol) and HATU (113.16 mg, 0.298 mmol)

- 88 039916 in DMF (4 mL) at room temperature for 1 hour gave the title compound (118 mg, 55% pure) as an off-white oil after purification by flash column chromatography (eluting with a gradient of 0-10% MeOH/ DXM). The resulting title compound was used in the next step without further purification. HPLC-MS (method H): [m/z]: 506.7 [M+H]+.

tert-Butyl yl]ethyl}carbamate (14)

N-(1H- 1,3-benzodiazol-2-ylmethyl)-N - {2-[4-(morpholine-4-carbonyl)-1,3-thiazol-2-

Similar to general procedure 6, 2-{2-[(1H-1,3-benzodiazol-2-ylmethyl)[(tert-butoxy)carbonyl]amino]ethyl}-1,3-thiazole-4-carboxylic acid (8) (99 .8 mg, 0.248 mmol), morpholine (25.93 mg, 0.298 mmol), DIPEA (96.16 mg, 0.744 mmol) and HATU (113.16 mg, 0.298 mmol) in DMF (4 ml) at room temperature in gave the title compound (110 mg) as an off-white oil over 1 hour after purification by flash column chromatography (0-10% MeOH/DCM gradient elution). The resulting title compound was used in the next step without further purification.

tert-Butyl N-(1H-1,3-benzodiazol-2-ylmethyl)-N-(2-{4-[methyl(phenyl)carbamoyl]-1,3-thiazol-2yl}ethyl)carbamate (15)

Similar to general procedure 6, butoxy)carbonyl]amino]ethyl}-1,3-thiazole-4-carboxylic

2-{2-[(1H-1,3-benzodiazol-2-ylmethyl)[(tert acid (8) (99.8 mg, 0.248 mmol), N-methylaniline (31.89 mg, 0.298 mmol), DIPEA (96 .16 mg, 0.744 mmol) and HATU (113.16 mg, 0.298 mmol) in

DMF (4 ml) at room temperature for 1 h gave the title compound (118 mg,

59% pure) as an off-white oil after purification by flash column chromatography (eluting with a 0-10% MeOH/DCM gradient). The resulting title compound was used in the next step without further purification.

HPLC-MS (method H): [m/z]: 492.7 [M+H] ⁺ .

tert-Butyl

N-(1H-1,3-benzodiazol-2-ylmethyl)-N-{2-[4-( {[2-(pyrrolidin-1 yl)phenyl]methyl}carbamoyl)-1,3-thiazol-2- yl]ethyl}carbamate (16)

Similar to general procedure 6, 2-{2-[(1H-1,3-benzodiazol-2-ylmethyl)[(tert-butoxy)carbonyl]amino]ethyl}-1,3-thiazole-4-carboxylic acid (8) (200 mg, 0.497 mmol), [3(pyrrolidin-1-yl)phenyl]methanamine (105 mg, 0.596 mmol), DIPEA (193 mg, 1.491 mmol) and HATU (227 mg, 0.596 mmol) in DMF (5 mL) at room temperature for 1 hour gave the title compound (100 mg, 30%, 84% pure) as an off-white oil after purification by flash column chromatography (eluting with a 0-10% MeOH/DCM gradient).

HPLC-MS (method H): [m/z]: 561.7 [M+H] ⁺ .

tert-Butyl N-(1H-1,3-benzodiazol-2-ylmethyl)-N-{2-[4-(dimethylcarbamoyl)-1,3-thiazol-2yl]ethyl}carbamate (17)

n

Similar to general procedure 6, 2-{2-[(1H-1,3-benzodiazol-2-ylmethyl)[(tert-butoxy)carbonyl]amino]ethyl}-1,3-thiazole-4-carboxylic acid (8) (100 mg, 0.248 mmol), dimethylamine (2 M solution in THF) (13 mg, 0.298 mmol), DIPEA (96 mg, 0.745 mmol) and HATU (113 mg, 0.298 mmol) in DMF (10 ml) at room temperature for 1 hour gave the title compound (90 mg, 67%, 80% pure) as a white solid after purification by flash column chromatography (eluting with a 0-10% MeOH/DCM gradient).

- 89 039916

HPLC-MS (method H): [m/z]: 430.6 [M+H]+.

tert-Butyl N-(1H-1,3-benzodiazol-2-ylmethyl)-N-(2-{4-[cyclohexyl(propan-2-yl)carbamoyl]-1,3thiazol-2-yl}ethyl)carbamate (eighteen)

Similar to general procedure 6, 2-{2-[(1H-1,3-benzodiazol-2-ylmethyl)[(tert-butoxy)carbonyl]amino]ethyl}-1,3-thiazole-4-carboxylic acid (8) (99 .8 mg, 0.248 mmol), K-(propan-2yl)cyclohexanamine (42.04 mg, 0.298 mmol), DIPEA (96.16 mg, 0.744 mmol) and HATU (113.16 mg, 0.298 mmol) in DMF ( 4 ml) at room temperature for 1 hour gave the title compound (124 mg, 17% pure) as an off-white oil after purification by flash column chromatography (eluting with a 0-10% MeOH/DCM gradient). The resulting title compound was used in the next step without further purification.

HPLC-MS (method H): [m/z]: 526.8 [M+H] ⁺ .

tert-Butyl N-(1H-1,3-benzodiazol-2-ylmethyl)-N-(2-{4-[(2-phenylpropαn-2-yl)carbamoyl]-1,3thiazol-2-yl}ethyl) carbamate (19)

Similar to general procedure 6, 2-{2-[(1H-1,3-benzodiazol-2-ylmethyl)[(tert-butoxy)carbonyl]amino]ethyl}-1,3-thiazole-4-carboxylic acid (8) (99 .8 mg, 0.248 mmol), 2-phenylpropan-2-amine (40.24 mg, 0.298 mmol), DIPEA (96.16 mg, 0.744 mmol) and HATU (113.16 mg, 0.298 mmol) in DMF (4 mL) at room temperature for 1 hour gave the title compound (120 mg, 51% pure) as an off-white oil after purification by flash column chromatography (eluting with a 0-10% MeOH/DCM gradient). The resulting title compound was used in the next step without further purification.

HPLC-MS (method H): [m/z]: 520.7 [M+H] ⁺ .

tert-Butyl N-(1H-1,3-benzodiazol-2-ylmethyl)-N-{2-[4-(benzylcarbamoyl)-1,3-thiazol-2yl]ethyl}carbamate (20)

n

Similar to general procedure 6, 2-{2-[(1H-1,3-benzodiazol-2-ylmethyl)[(tert-butoxy)carbonyl]amino]ethyl}-1,3-thiazole-4-carboxylic acid (8) (350 mg, 0.87 mmol), phenylmethanamine (103 mg, 0.957 mmol), DIPEA (337 mg, 2.61 mmol) and HATU (397 mg, 1.04 mmol) in DMF (10 mL) gave the title compound ( 390 mg, 89% pure) as a white solid after purification by flash column chromatography (eluting with a 0-10% MeOH/DCM gradient).

HPLC-MS (method H): [m/z]: 492.6 [M+H] ⁺ .

tert-Butyl N-{2-[4-(benzylcarbamoyl)-1,3-thiazol-2-yl]ethyl}-N-[(1-methyl-1H-1,3-benzodiazol-2yl)methyl]carbamate ( 21)

/

Into a stirred solution of tert-butyl N-(1H-1,3-benzodiazol-2-ylmethyl)-N-{2-[4(benzylcarbamoyl)-1,3-thiazol-2-yl]ethyl}carbamate (20) ( 380 mg, 0.773 mmol) and TEA (78 mg, 0.773 mmol) in DCM (15 mL) MeI (165 mg, 1.159 mmol) was added under argon. The reaction mixture was stirred at room temperature overnight. The reaction mixture was evaporated to dryness in vacuo to give the title compound (280 mg, 72% pure) as an off-white solid. The crude product was used in the next step without purification.

HPLC-MS (method H): [m/z]: 506.6 [M+H] ⁺ .

tert-Butyl N-(1H-1,3-benzodiazol-2-ylmethyl)-N-(2-{4-[(pyridin-3-ylmethyl)carbamoyl]-1,3-thiazol2-yl}ethyl)carbamate ( 22)

- 90 039916

Similar to general procedure 6, 2-{2-[(1H-1,3-benzodiazol-2-ylmethyl)[(tert-butoxy)carbonyl]amino]ethyl}-1,3-thiazole-4-carboxylic acid (8) (199 .6 mg, 0.496 mmol), pyridin-3ylmethanamine (59 mg, 0.546 mmol), DIPEA (192.3 mg, 1.488 mmol) and HATU (226 mg, 0.595 mmol) in DMF (8 mL) gave the title compound ( 184 mg, 75%) as a white solid after purification by flash column chromatography (eluting with a 0-10% MeOH/DCM gradient).

1H NMR (CDCl ₃ , 400 MHz): δ [ppm] = 8.61 (d, J=1.6 Hz, 1H), 8.52 (d, J=3.6 Hz, 1H), 7.96 (s, 1H), 7.81 (s, 1H), 7.73-7.67 (m, 1H), 7.55 (dd, J=6.0, 3.2 Hz, 2H) , 7.28 (s, 1H), 7.25 (dd, J=6.1, 3.2 Hz, 2H), 4.63 (d, J=6.6 Hz, 4H), 3.77 ( t, J=6.5 Hz, 2H), 3.22 (t, J=6.3 Hz, 2H), 1.37 (s, 9H).

HPLC-MS (method H): [m/z]: 493.4 [M+H]+.

tert-Butyl N-(1H-1,3-benzodiazol-2-ylmethyl)-N-(2-{4-[(pyridin-4-ylmethyl)carbamoyl]-1,3-thiazol2-yl}ethyl)carbamate ( 23)

n

Similar to general procedure 6, 2-{2-[(1H-1,3-benzodiazol-2-ylmethyl)[(tert-butoxy)carbonyl]amino]ethyl}-1,3-thiazole-4-carboxylic acid (8) (199 .6 mg, 0.496 mmol), pyridin-4ylmethanamine (59 mg, 0.546 mmol), DIPEA (192.3 mg, 1.488 mmol) and HATU (226 mg, 0.595 mmol) in DMF (8 mL) gave the title compound ( 140 mg, 57%) as a white solid after purification by flash column chromatography (eluting with a 0-10% MeOH/DCM gradient).

1H NMR (CDCl ₃ , 400 MHz): δ [ppm] = 8.56 (d, J=5.9 Hz, 2H), 7.98 (s, 1H), 7.82 (s, 1H ), 7.59-7.48 (m, 2H), 7.24 (dd, J=6.0, 3.2 Hz, 4H), 4.62 (d, J=6.6 Hz, 4H) , 3.78 (t, J=6.5 Hz, 2H), 3.25 (t, J=6.4 Hz, 2H), 1.41 (d, J=13.9 Hz, 9H).

HPLC-MS (method H): [m/z]: 493.4 [M+H] ⁺ .

tert-Butyl N-(1H-1,3-benzodiazol-2-ylmethyl)-K - {2-[4-({[3-(trifluoromethyl)pyridin-2yl]methyl}carbamoyl)-1,3-thiazol- 2-yl]ethyl}carbamate (24)

Similar to general procedure 6, 2-{2-[(1H-1,3-benzodiazol-2-ylmethyl)[(tert-butoxy)carbonyl]amino]ethyl}-1,3-thiazole-4-carboxylic acid (8) (80 .09 mg, 0.199 mmol), [3(trifluoromethyl)pyridin-2-yl]methanamine hydrochloride (46.54 mg, 0.219 mmol), DIPEA (102.9 mg, 0.796 mmol), and HATU (90.8 mg, 0.239 mmol) in DMF (2.5 mL) gave the title compound (85 mg, 76%, 98% pure) as a white solid after purification by flash column chromatography (eluting with a 0-10% MeOH/DCM gradient).

HPLC-MS (method H): [m/z]: 561.5 [M+H] ⁺ .

tert-Butyl N-(1H-1,3-benzodiazol-2-ylmethyl)-N-(2-{4-[(5,6,7,8-tetrahydroquinolin-8yl)carbamoyl]-1,3-thiazol- 2-yl}ethyl)carbamate (25)

n

Similar to general procedure 6, 2-{2-[(1H-1,3-benzodiazol-2-ylmethyl)[(tert-butoxy)carbonyl]amino]ethyl}-1,3-thiazole-4-carboxylic acid (8) (80 09 mg, 0.199 mmol), N-methyl 5,6,7,8-tetrahydroquinoline-8-amine dihydrochloride (48.4 mg, 0.219 mmol), DIPEA (102.9 mg, 0.796 mmol), and HATU (90.8 mg, 0.239 mmol) in DMF (2.5 mL) gave the title compound (92 mg, 87%) as a white solid after purification by flash column chromatography (eluting with a 0-10% MeOH/DCM gradient).

HPLC-MS (method H): [m/z]: 533.5 [M+H] ⁺ .

tert-Butyl N-(1H-1,3-benzodiazol-2-ylmethyl)-N-{2-[4-({5H,6H,7H-cyclopenta[b]pyridin-7yl}carbamoyl)-1,3- thiazol-2-yl]ethyl}carbamate (26)

- 91 039916

n

Similar to general procedure 6, 2-{2-[(1H-1,3-benzodiazol-2-ylmethyl)[(tert-butoxy)carbonyl]amino]ethyl}-1,3-thiazole-4-carboxylic acid (8) (80 .9 mg, 0.199 mmol), N-methyl5H,6H,7H-cyclopenta[b]pyridine-7-amine hydrochloride (37.35 mg, 0.219 mmol), DIPEA (102.9 mg, 0.796 mmol), and HATU (90 .8 mg, 0.239 mmol) in DMF (2.5 ml) gave the title compound (90 mg, 87%) as a white solid after purification by flash column chromatography (eluting with a gradient of 0-10% MeOH/DCM ).

HPLC-MS (method H): [m/z]: 519.5 [M+H]+.

tert-Butyl N-(1H-1,3-benzodiazol-2-ylmethyl)-N-[2-(4-{[(4-methylmorpholin-2-yl)methyl]carbamoyl}-1,3-thiazol-2 -yl)ethyl]carbamate (27)

n

Similar to general procedure 6, 2-{2-[(1H-1,3-benzodiazol-2-ylmethyl)[(tert-butoxy)carbonyl]amino]ethyl}-1,3-thiazole-4-carboxylic acid (8) (3 g, 7.545 mmol), (3-fluoropyridin2-yl)methanamine dihydrochloride (A2) (2.26 g, 11.18 mmol), DIPEA (12.98 ml, 74.54 mmol) and HATU (4.251 g, 11. 18 mmol) in DMF (60 ml) gave the title compound (4.13 mg, 89%) as a yellow oil after purification by flash column chromatography (kp-NH, eluting with a gradient of 20100% EtOAc/heptane, then 0 -20% MeOH/EtOAc).

¹ H NMR (DMSO-d ₆ , 500 MHz): δ [ppm] = 12.29 (s, 1H), 8.69 (s, 1H), 8.36 (s, 1H), 8, 17 (s, 1H), 7.70 (t, J=9.5 Hz, 1H), 7.48 (s, 2H), 7.40 (dt, J=8.6, 4.4 Hz, 1H ), 7.14 (s, 2H), 4.66 (d, J=8.8 Hz, 4H), 3.73 (s, 2H), 2.52 (s, 2H), 1.99 (s , 4H), 1.26 (d, J=44.9 Hz, 9H).

HPLC-MS (method A): [m/z]: 511.15 [M+H] ⁺ .

tert-Butyl N-(1H-1,3-benzodiazol-2-ylmethyl)-N-[2-(4-{[(4-methylmorpholin-2yl)methyl]carbamoyl}-1,3-thiazol-2-yl )ethyl]carbamate (28)

n

Similar to general procedure 6, 2-{2-[(1H-1,3-benzodiazol-2-ylmethyl)[(tert-butoxy)carbonyl]amino]ethyl}-1,3-thiazole-4-carboxylic acid (8) (99 .8 mg, 0.248 mmol), (4methylmorpholin-3-yl)methanamine (35.5 mg, 0.273 mmol), DIPEA (96.16 mg, 0.744 mmol) and T3P (189.4 mg, 0.298 mmol) in DMF ( 4 ml) gave the title compound (90 mg, 70%) as a white solid after purification by flash column chromatography (eluting with a 0-10% MeOH/DCM gradient).

1H NMR (CDCl ₃ , 400 MHz): δ [ppm] = 7.90 (s, 1H), 7.59 (s, 3H), 7.28 (t, J=3.6 Hz, 1H ), 4.64 (s, 2H), 3.79 (td, J=12.0, 4.7 Hz, 5H), 3.70-3.59 (m, 2H), 3.47 (ddd, J=14.7, 10.3, 6.6 Hz, 4H), 3.22 (t, J=6.4 Hz, 2H), 2.74 (d, J=11.4 Hz, 1H), 2.49-2.32 (m, 6H), 1.40 (s, 9H).

tert-Butyl N-(1H-1,3-benzodiazol-2-ylmethyl)-N-[2-(4-{[(6-methyl-pyridin-2-yl)methyl]carbamoyl}1,3-thiazol-2- yl)ethyl]carbamate (29)

Similar to general procedure 6, 2-{2-[(1H-1,3-benzodiazol-2-ylmethyl)[(tert-butoxy)carbonyl]amino]ethyl}-1,3-thiazole-4-carboxylic acid 6 (99.8 mg, 0.248 mmol), (6methylpyridin-2-yl)methanamine (33.33 mg, 0.273 mmol), DIPEA (96.16 mg, 0.744 mmol) and T3P (189.4 mg, 0.298 mmol) in DMF (4 ml ) gave the title compound (95 mg, 75%) as a white solid after purification by flash column chromatography (eluting with a 0-10% MeOH/DCM gradient).

1H NMR (CDCl3, 400 MHz): δ [ppm] = 8.14 (s, 1H), 7.90 (s, 1H), 7.55 (dd, J=14.1, 6.6 Hz, 3H), 7.23 (dd, J=6.0, 3.2 Hz, 2H), 7.14 (d, J=7.7 Hz, 1H), 7.06 (d, J=7 .7 Hz, 1H), 4.71 (d, J=5.5 Hz, 2H), 4.66 (s, 2H), 3.80 (t,

- 92 039916

J=6.3Hz, 2H), 3.22(t, J=6.4Hz, 2H), 2.55(s, 3H), 1.34(s, 9H).

tert-Butyl N-(1H-1,3-benzodiazol-2-ylmethyl)-N-[2-(4-{[(5-fluoropyridin-2-yl)methyl]carbamoyl}1,3-thiazol-2- yl)ethyl]carbamate (30)

Similar to general procedure 6, 2-{2-[(1H-1,3-benzodiazol-2-ylmethyl)[(tert-butoxy)carbonyl]amino]ethyl}-1,3-thiazole-4-carboxylic acid (8) (99 .8 mg, 0.248 mmol), (5fluoropyridin-2-yl)methanamine (34.41 mg, 0.273 mmol), DIPEA (96.16 mg, 0.744 mmol) and T3P (189.4 mg, 0.298 mmol) in DMF ( 4 ml) gave the title compound (89 mg, 70%) as a white solid after purification by flash column chromatography (eluting with a 0-10% MeOH/DCM gradient).

¹ H NMR (CDCl3, 400 MHz): δ [ppm] = 8.44 (s, 1H), 8.12 (s, 1H), 7.92 (s, 1H), 7.56 (s , 2H), 7.44-7.30 (m, 2H), 7.25 (dd, J=6.1, 3.2 Hz, 2H), 4.73 (d, J=5.6 Hz, 2H), 4.66 (s, 2H), 3.79 (t, J=6.3Hz, 2H), 3.23 (t, J=6.3Hz, 2H), 3.04 (s, 1H ), 1.34 (s, 9H).

tert-Butyl N-(1H-1,3-benzodiazol-2-ylmethyl)-N-(2-{4-[(pyrimidin-4-ylmethyl)carbamoyl]-1,3thiazol-2-yl}ethyl)carbamate ( 31)

Similar to general procedure 6, 2-{2-[(1H-1,3-benzodiazol-2-ylmethyl)[(tert-butoxy)carbonyl]amino]ethyl}-1,3-thiazole-4-carboxylic acid (8) (150 mg, 0.373 mmol), pyrimidin-4ylmethanamine (48.8 mg, 0.447 mmol), DIPEA (48.1 mg, 0.373 mmol) and HATU (141.7 mg, 0.373 mmol) in DMF (2 ml) at room temperature in overnight gave the title compound (80 mg, 60% pure) as a yellow oil after purification by flash column chromatography (gradient elution with 10% MeOH in DCM).

HPLC-MS (method H): [m/z]: 494.6 [M+H]+.

tert-Butyl N-(1H-1,3-benzodiazol-2-ylmethyl)-N-[2-(4-{[(5-methoxypyridin-2yl)methyl]carbamoyl}-1,3-thiazol-2-yl )ethyl]carbamate (32)

n

Similar to general procedure 6, 2-{2-[(1H-1,3-benzodiazol-2-ylmethyl)[(tert-butoxy)carbonyl]amino]ethyl}-1,3-thiazole-4-carboxylic acid (8) (150 mg, 0.373 mmol), (5methoxypyridin-2-yl)methanamine (48.17 mg, 0.447 mmol), DIPEA (48.1 mg, 0.373 mmol) and HATU (141.7 mg, 0.373 mmol) in DMF (2 ml ) at room temperature overnight gave the title compound (80 mg, 41%) as a brown solid after purification by flash column chromatography (gradient elution with 10% MeOH in DCM).

HPLC-MS (method H): [m/z]: 523.6 [M+H] ⁺ .

tert-Butyl N-(1H-1,3-benzodiazol-2-ylmethyl)-N-(2-{4-[(pyrazin-2-ylmethyl)carbamoyl]-1,3-thiazol2-yl}ethyl)carbamate ( 33)

n

Similar to general procedure 6, 2-{2-[(1H-1,3-benzodiazol-2-ylmethyl)[(tert-butoxy)carbonyl]amino]ethyl}-1,3-thiazole-4-carboxylic acid (8) (150 mg, 0.373 mmol), pyrazin-2ylmethanamine (48.8 mg, 0.447 mmol), DIPEA (192.68 mg, 1.491 mmol) and HATU (141.7 mg, 0.373 mmol) in DMF (2 ml) at room temperature in overnight gave the title compound (95 mg, 52%) as a yellow solid after purification by flash column chromatography (gradient elution with 10% MeOH in DCM).

HPLC-MS (method H): [m/z]: 394.5 [M+H-Boc] ⁺ .

tert-Butyl N-(1H-1,3-benzodiazol-2-ylmethyl)-N-[2-(4-{[(6-oxo-1,6-dihydropyridin-2-93 039916 yl)methyl]carbamoyl} -1,3-thiazol-2-yl)ethyl]carbamate (34)

Similar to general procedure 6, 2-{2-[(1H-1,3-benzodiazol-2-ylmethyl)[(tert-butoxy)carbonyl]amino]ethyl}-1,3-thiazole-4-carboxylic acid (8) (150 mg, 0.373 mmol), 6(aminomethyl)-1,2-dihydropyridin-2-one (55.52 mg, 0.447 mmol), DIPEA (48.17 mg, 0.373 mmol) and HATU (141.7 mg, 0.373 mmol ) in DMF (2 mL) at room temperature overnight gave the title compound (90 mg, 47%) as a yellow solid after purification by flash column chromatography (eluting with a 10% MeOH/DCM gradient).

HPLC-MS (method H ): [m/z]: 509.6 [M+H]+.

tert-Butyl N(1H-1,3-benzodiazol-2-ylmethyl)-N[2-(4-{[(6-carbamoylpyridin-3yl)methyl]carbamoyl}-1,3-thiazol-2-yl)ethyl ]carbamate (35) and tert-butyl N(1H-1,3-benzodiazol-2ylmethyl)-N[2-(4-{[(6-cyanopyridin-3-yl)methyl]carbamoyl}-1,3-thiazole -2-yl)ethyl]carbamate (36)

Similar to general procedure 6, 2-{2-[(1H-1,3-benzodiazol-2-ylmethyl)[(tert-butoxy)carbonyl]amino]ethyl}-1,3-thiazole-4-carboxylic acid (8) (100 mg, 0.248 mmol), 5(aminomethyl)pyridine-2-carbonitrile (33 mg, 0.248 mmol), HATU (189 mg, 0.497 mmol) and DIPEA (96 mg, 0.745 mmol) in DMF (1 ml) at room temperature in for 18 hours, gave a 2:1 ratio of bocamide and boc-nitrile (80 mg) after purification by flash column chromatography (DCM:MeOH, 9:1). The resulting mixture was used in the next step without separation.

HPLC-MS (method H): [m/z]: 418.5 [M+H-boc] ⁺ and 436.3 [M+H-boc] ⁺ .

tert-Butyl X-(1H-1,3-benzodiazol-2-ylmethyl)-N-[2-(4-{[(3,5-dimethylpyridin-2yl)methyl]carbamoyl}-1,3-thiazol-2 -yl)ethyl]carbamate (37)

Similar to general procedure 6, 2-{2-[(1H-1,3-benzodiazol-2-ylmethyl)[(tert-butoxy)carbonyl]amino]ethyl}-1,3-thiazole-4-carboxylic acid (8) (0 .3 g, 0.708 mmol), (3,5dimethylpyridin-2-yl)methanamine hydrochloride (0.183 g, 1.062 mmol), DIPEA (0.555 ml, 3.187 mmol) and HATU (0.404 g, 1.062 mmol) in DMF (6 ml) at room temperature for 4 h gave the title compound (0.198 g, 51%) as a yellow oil after purification by flash column chromatography (kp-NH, eluting with an EtOAc (30%)/heptane (70%) gradient, then 100 % EtOAc).

¹ H NMR (DMSO-d ₆ , 500 MHz): δ [ppm] = 12.29 (s, 1H), 8.75 (s, 1H), 8.19 (s, 2H), 7, 56 (d, J=7.6 Hz, 1H), 7.48-7.40 (m, 2H), 7.14 (p, J=7.0 Hz, 2H), 4.66 (s, 2H ), 4.53 (d, J=4.8 Hz, 2H), 3.73 (s, 2H), 2.27 (s, 3H), 2.23 (s, 3H), 1.31 (s , 9H).

HPLC-MS (method A): [m/z]: 521.15 [M+H] ⁺ .

tert-Butyl N-(1H-1,3-benzodiazol-2-ylmethyl)-N-(2-{4-[(pyrimidin-2-ylmethyl)carbamoyl]-1,3thiazol-2-yl}ethyl)carbamate ( 38)

Similar to general procedure 6, 2-{2-[(1H-1,3-benzodiazol-2-ulmethyl)[(tert-butoxy)carbonyl]amino]ethyl}-1,3-thiazole-4-carboxylic acid (8) (100 mg, 0.20 mmol), 1-(pyrimidin2-yl)methanamine (22 mg, 0.20 mmol), DIPEA (0.1 ml, 0.60 mmol) and HATU (113 mg, 0.30 mmol) in DCM (5 mL) gave the title compound (86 mg, 73%) as a brown residue after purification by flash column chromatography (eluting with a 0-20% MeOH/EtOAc gradient).

HPLC-MS (Method A): [m/z]: 494.1 [M+H] ⁺ .

tert-Butyl N-(1H-1,3-benzodiazol-2-ylmethyl)-N-{2-[4-({[2-(4-methylpiperazin-1-yl)phenyl]methyl}carbamoyl)-1, 3-thiazol-2-yl]ethyl}carbamate (39)

- 94 039916

Similar to General Procedure 6, 2-{2-[(1H-1,3-Benzodiazol-2-ylmethyl)[(tert-butoxy)carbonyl]amino]ethyl}-1,3-thiazole-4-carboxylic acid (8) (130mg , 0.24 mmol, 75% pure), 1[2-(4-methylpiperazin-1-yl)phenyl]methanamine (75 mg, 0.36 mmol), DIPEA (127 µl, 0.73 mmol), and HATU ( 138 mg, 0.36 mmol) in DMF (2 mL) gave the title compound (13 mg, 9%) as a white solid after purification by basic prep-HPLC.

HPLC-MS (Method D): [m/z]: 590.3 [M+H]+.

tert-Butyl N-(1H-1,3-benzodiazol-2-ylmethyl)-N-[2-(4-{[(2,6-difluorophenyl)methyl]carbamoyl}-1,3thiazol-2-yl)ethyl ]carbamate (40)

n

Similar to general procedure 6, 2-{2-[(1H-1,3-benzodiazol-2-ylmethyl)[(tert-butoxy)carbonyl]amino]ethyl}-1,3-thiazole-4-carboxylic acid (8) (110 mg, 0.25 mmol, 90% pure), 1(2,6-difluorophenyl)methanamine (53 mg, 0.37 mmol), DIPEA (0.13 ml, 0.74 mmol) and HATU (140 mg, 0 .37 mmol) in DMF (2 mL) gave the title compound (90 mg, 68%) as a yellow oil after purification by flash column chromatography (kp-NH, eluting with a 2-100% EtOAc/heptane gradient).

HPLC-MS (Method E): [m/z]: 528.3 [M+H] ⁺ .

tert-Butyl N-(1H-1,3-benzodiazol-2-ylmethyl)-N-{2-[4-({[2-(dimethylamino)phenyl]methyl}carbamoyl)-1,3-thiazol-2- yl]ethyl}carbamate (41)

n

Similar to general procedure 6, 2-{2-[(1H-1,3-benzodiazol-2-ylmethyl)[(tert-butoxy)carbonyl]amino]ethyl}-1,3-thiazole-4-carboxylic acid (8) (109 mg, 0.22 mmol, 80% purity), 2(aminomethyl)-N,N-dimethylaniline (66 mg, 0.44 mmol), DIPEA (226 μl, 1.30 mmol) and HATU (240 mg,

0.64 mmol) in DMF (2 mL) at 50° C. gave the title compound (73 mg, 61%) as an orange oil after purification by flash column chromatography (kp-NH, eluting with a 5-100% gradient). EtOAc/heptane).

HPLC-MS (Method D): [m/z]: 535.2 [M+H] ⁺ .

tert-Butyl N-(1H-1,3-benzodiazol-2-ylmethyl)-N-[2-(4-{[(2-cyanophenyl)methyl]carbamoyl}-1,3thiazol-2-yl)ethyl]carbamate (42)

n

Similar to general procedure 6, 2-{2-[(1H-1,3-benzodiazol-2-ylmethyl)[(tert-butoxy)carbonyl]amino]ethyl}-1,3-thiazole-4-carboxylic acid (8) (109 mg, 0.22 mmol, 80% pure), 2(aminomethyl)benzonitrile hydrochloride (74 mg, 0.44 mmol), DIPEA (226 µl, 1.30 mmol) and HATU (240 mg, 0.64 mmol ) in DMF (2 mL) at 50°C gave the crude title compound (54 mg, 30%, 63% pure) as an orange oil after purification by flash column chromatography (kp-NH, eluting with a gradient of 5-100 % EtOAc/heptane).

HPLC-MS (Method D): [m/z]: 517.2 [M+H] ⁺ .

tert-Butyl N-(1H-1,3-benzodiazol-2-ylmethyl)-N-{2-[4-({[2-(trifluoromethoxy)phenyl]methyl}carbamoyl)-1,3-thiazol-2- yl]ethyl}carbamate (43)

- 95 039916

Similar to general procedure 6, 2-{2-[(III-1,3-benzodiazol-2-ylmethyl)[(tert-butoxy)carbonyl]amino]ethyl}-1,3-thiazole-4-carboxylic acid (8) (109 mg, 0.22 mmol, 80% pure), 1-[2(trifluoromethoxy)phenyl]methanamine (103 mg, 0.54 mmol), DIPEA (283 μl, 1.63 mmol) and HATU (248 mg, 0.65 mmol) in DMF (2 mL) at 50°C gave the crude title compound (110 mg, 78%, 88% pure) as a yellow oil after purification by flash column chromatography (kp-NH, eluting in gradient 8-100% EtOAc/heptane).

HPLC-MS (Method E): [m/z]: 576.2 [M+H]+.

tert-Butyl N-(1H-1,3-benzodiazol-2-ylmethyl)-M-(2-{4-[(1-phenylethyl)carbamoyl]-1,3-thiazol-2yl}ethyl)carbamate (44)

Similar to general procedure 6, 2-{2-[(1H-1,3-benzodiazol-2-ylmethyl)[(tert-butoxy)carbonyl]amino]ethyl}-1,3-thiazole-4-carboxylic acid (8) (0.11 g, 0.22 mmol, 80% pure), 1-phenylethanamine (0.07 ml, 0.54 mmol), DIPEA (0.303 ml, 1.63 mmol) and HATU (0.25 g, 0 .64 mmol) in DMF (2 mL) gave the crude title compound (110 mg, 77%, 77% pure) as a yellow oil after purification by flash column chromatography (KP-NH, eluting with a gradient of 8-100% EtOAc/heptane).

HPLC-MS (Method E): [m/z]: 506.2 [M+H] ⁺ .

tert-Butyl N-(1H-1,3-benzodiazol-2-ylmethyl)-N-{2-[4-({[2-(difluoromethoxy)phenyl]methyl}carbamoyl)-1,3-thiazol-2- yl]ethyl}carbamate (45)

Similar to general procedure 6, 2-{2-[(1H-1,3-benzodiazol-2-ylmethyl)[(tert-butoxy)carbonyl]amino]ethyl}-1,3-thiazole-4-carboxylic acid (8) (109 mg, 0.22 mmol, 80% pure), 1-[2(difluoromethoxy)phenyl]methanamine (83 mg, 0.48 mmol), DIPEA (0.23 ml, 1.3 mmol), and HATU (250 mg, 0.65 mmol) in DMF (2 mL) gave the crude title compound (470 mg) as an orange oil which was used in the next step without purification.

HPLC-MS (method A): [m/z]: 558.25 [M+H] ⁺ .

tert-Butyl N-(1H-1,3-benzodiazol-2-ylmethyl)-N-{2-[4-({[2-(morpholine-4sulfonyl)phenyl]methyl}carbamoyl)-1,3-thiazol- 2-yl]ethyl}carbamate (46)

Similar to general procedure 6, 2-{2-[(1H-1,3-benzodiazol-2-ylmethyl)[(tert-butoxy)carbonyl]amino]ethyl}-1,3-thiazole-4-carboxylic acid (8) (109 mg, 0.217 mmol, 80% pure), 1-[2-(morpholin-4-ylsulfonyl)phenyl]methanamine hydrochloride (140 mg, 0.48 mmol), DIPEA (0.23 ml, 1.3 mmol) and HATU (247 mg, 0.65 mmol) in DMF (2 mL) gave the crude title compound (440 mg) as an orange oil after the reaction mixture was evaporated in vacuo. The resulting product was used without further purification.

HPLC-MS (method A): [m/z]: 641.35 [M+H] ⁺ .

tert-Butyl N-(1H-1,3-benzodiazol-2-ylmethyl)-N-[2-(4-{[2-(pyridin-2-yl)ethyl]carbamoyl}-1,3thiazol-2-yl )ethyl]carbamate (47)

- 96 039916

Similar to general procedure 6, 2-{2-[(1H-1,3-benzodiazol-2-ylmethyl)[(tert-butoxy)carbonyl]amino]ethyl}-1,3-thiazole-4-carboxylic acid (8) (100 mg, 0.2 mmol, 80% purity), 2(pyridin-2-yl)ethanamine (49 mg, 0.4 mmol), DIPEA (104 µl, 0.6 mmol) and HATU (151 mg, 0.4 mmol) in DMF (2 mL) gave the title compound (52 mg, 52%) as an off-white solid after purification by KP-NH flash column chromatography, eluting with a 5100% EtOAc/heptane gradient).

HPLC-MS (Method A): [m/z]: 507.15 [M+H]+.

tert-Butyl N-(1H-1,3-benzodiazol-2-ylmethyl)-N-(2-{4-[(3-fluoropyridin-2-yl)carbamoyl]-1,3thiazol-2-yl}ethyl) carbamate (48)

Similar to general procedure 6, a solution of 2-{2-[(1H-1,3-benzodiazol-2-ylmethyl)[(tert-butoxy)carbonyl]amino]ethyl}-1,3-thiazole-4-carboxylic acid (8) ( 80%, 150 mg, 0.3 mmol), 3-fluoropyridine-2-amine (100 mg, 0.89 mmol), DIPEA (312 µl, 1.78 mmol) and HATU (340 mg, 0.87 mmol) in DMF (2 ml) was heated at 100° C. for 16 h. The reaction mixture was evaporated in vacuo to give the crude title compound (705 mg) as a brown oil which was used in the next step without purification.

HPLC-MS (method A): [m/z]: 497.10 [M+H] ⁺ .

tert-Butyl N-(1H-1,3-benzodiazol-2-ylmethyl)-N-(2-{4-[(2-phenylethyl)carbamoyl]-1,3-thiazol-2yl}ethyl)carbamate (49)

Similar to general procedure 6, 2-{2-[(1H-1,3-benzodiazol-2-ylmethyl)[(tert-butoxy)carbonyl]amino]ethyl}-1,3-thiazole-4-carboxylic acid (8) (0 .1 g, 0.25 mmol), 2-phenylethanamine (0.03 ml, 0.25 mmol), DIPEA (0.13 ml, 0.75 mmol) and HATU (0.14 g, 0.37 mmol) in DMF (2 mL) gave the title compound (71 mg, 56%) as a yellow oil after purification by flash column chromatography (eluting with a 2-100% EtOAc/heptane gradient).

HPLC-MS (method A): [m/z]: 506.2 [M+H] ⁺ .

tert-Butyl N-(1H-1,3-benzodiazol-2-ylmethyl)-N-[2-(4-{[(3-fluoro-6-methylpyridin-2yl)methyl]carbamoyl}-1,3-thiazole -2-yl)ethyl]carbamate (50)

Similar to general procedure 6, 2-{2-[(1H-1,3-benzodiazol-2-ylmethyl)[(tert-butoxy)carbonyl]amino]ethyl}-1,3-thiazole-4-carboxylic acid (8) (150 mg, 0.298 mmol, 80% pure), (3-fluoro-6-methylpyridin-2-yl)methanamine hydrochloride (79 mg, 0.447 mmol), DIPEA (156 µl, 0.894 mmol) and HATU (230 mg, 0.596 mmol) in DMF (3 mL) gave the title compound (76 mg, 48%) as a pale yellow oil after purification by flash column chromatography (kp-NH, eluting with a 0-100% EtOAc/heptane gradient).

HPLC-MS (method A): [m/z]: 525.40 [M+H] ⁺ .

tert-Butyl N-(1H-1,3-benzodiazol-2-ylmethyl)-N-[2-(4-{[1-(pyridin-2-yl)ethyl]carbamoyl}-1,3thiazol-2-yl )ethyl]carbamate (51)

Similar to general procedure 6, 2-{2-[(1H-1,3-benzodiazol-2-ylmethyl)[(tert-butoxy)carbonyl]amino]ethyl}-1,3-thiazole-4-carboxylic acid (8) (150 mg, 0.298 mmol, 80% purity), 1-(pyridin-2-yl)ethanamine (55 mg, 0.447 mmol), DIPEA (156 µl, 0.894 mmol) and HATU (227 mg, 0.596 mmol) in DMF (3 ml ) gave the title compound (78 mg, 50%) as a colorless oil after purification by flash column chromatography (kp-NH, eluting with a 0-100% EtOAc/heptane gradient).

HPLC-MS (method A): [m/z]: 507.20 [M+H] ⁺ .

- 97 039916 tert-Butyl N-(1H-1,3-benzodiazol-2-ylmethyl)-N-{2-[4-({[6-(trifluoromethyl)pyridin-3yl]methyl}carbamoyl)-1,3 -thiazol-2-yl]ethyl}carbamate (52)

n

Similar to general procedure 6, 2-{2-[(1H-1,3-benzodiazol-2-ylmethyl)[(tert-butoxy)carbonyl]amino]ethyl}-1,3-thiazole-4-carboxylic acid (8) (150 mg, 0.298 mmol, 80% pure), 1-[6(trifluoromethyl)pyridin-3-yl]methanamine (79 mg, 0.447 mmol), DIPEA (156 μL, 0.894 mmol) and HATU (227 mg, 0.596 mmol ) in DMF (3 mL) gave the title compound (92 mg, 46%) as a colorless oil after purification by flash column chromatography (kp-NH, eluting with a 0-100% EtOAc/heptane gradient).

HPLC-MS (Method A): [m/z]: 561.35 [M+H]+.

tert-Butyl K-(1H-1,3-benzodiazol-2-ylmethyl)-K-[2-(4-{[(3-chloropyridin-2-yl)methyl]carbamoyl}1,3-thiazol-2- yl)ethyl]carbamate (53)

Similar to general procedure 6, 2-{2-[(1H-1,3-benzodiazol-2-ylmethyl)[(tert-butoxy)carbonyl]amino]ethyl}-1,3-thiazole-4-carboxylic acid (8) (150 mg, 0.34 mmol, 92% pure), 1-(3-chloropyridin-2yl)methanamine dihydrochloride (111 mg, 0.51 mmol), DIPEA (299 µl, 1.71 mmol) and HATU (196 mg , 0.51 mmol) in DMF (2 mL) gave the title compound (161 mg, 73% pure, 63%) as a yellow oil after purification by flash column chromatography (kp-NH, eluting with a gradient of 20-100% EtOAc /heptane).

¹ H NMR (CDCl ₃ , 500 MHz): δ [ppm] = 10.47 (s, 1H), 8.59 (s, 1H), 8.52-8.42 (m, 1H), 7.92 (s, 1H), 7.71 (d, J=8.2 Hz, 2H), 7.32 (s, 1H), 7.25-7.16 (m, 3H), 4.85 (d, J=4.8 Hz, 2H), 4.69 (s, 2H), 3.81 (s, 2H), 3.23 (t, J=6.5 Hz, 2H), 1.35 (s, 9H).

HPLC-MS (method A): [m/z]: 527.35 [M+H] ⁺ .

tert-Butyl N-(1H-1,3-benzodiazol-2-yl-methyl)-N-{2-[4-({[2-(tert-butoxy)pyridin-3yl]methyl}carbamoyl)-1 ,3-thiazol-2-yl]ethyl}carbamate (54)

Similar to general procedure 6, 2-{2-[(1H-1,3-benzodiazol-2-ylmethyl)[(tert-butoxy)carbonyl]amino]ethyl}-1,3-thiazole-4-carboxylic acid (8) (150 mg, 0.34 mmol, 92% pure), 1(2-tert-butoxypyridin-3-yl)methanamine (93 mg, 0.514 mmol), DIPEA (179 µl, 1.03 mmol) and HATU (196 mg, 0 .51 mmol) in DMF (2 mL) gave the title compound (205 mg, 61%, 58% pure) as a yellow oil after purification by flash column chromatography (kp-NH, eluting with a gradient of 20-100% EtOAc /heptane).

1H NMR (CDCl3, 500 MHz): δ [ppm] = 10.09 (s, 1H), 7.89 (d, J=15.7 Hz, 2H), 7.71 (s, 1H) , 7.54-7.45 (m, 2H), 7.40 (d, J=4.9 Hz, 1H), 7.24 (s, 1H), 6.77 (td, J=7.3 , 5.0 Hz, 2H), 4.60 (s, 2H), 4.49 (d, J=6.5 Hz, 2H), 3.77 (t, J=6.5 Hz, 2H), 3.22 (s, 2H), 1.63 (s, 9H), 1.33 (s, 9H).

HPLC-MS (method A): [m/z]: 565.15 [M+H] ⁺ .

tert-Butyl N-(1H-1,3-benzodiazol-2-ylmethyl)-N-[2-(4-{[(1-methyl-1 H-imidazol-5-yl)methyl]carbamoyl}-1, 3-thiazol-2-yl)ethyl] carbamate (55)

n

Similar to general procedure 6, 2-{2-[(1H-1,3-benzodiazol-2-ylmethyl)[(tert-butoxy)carbonyl]amino]ethyl}-1,3-thiazole-4-carboxylic acid (8) (150 mg, 0.37 mmol), (1-methyl-1Himidazol-5-yl)methanamine (62 mg, 0.56 mmol), DIPEA (185 µl, 1.12 mmol) and HATU (213 mg, 0.56 mmol ) in DMF (2 mL) gave the title compound (175 mg, 95%) as a yellow oil after purification by flash column chromatography (eluting with a 0-3% MeOH/DCM gradient).

1H NMR (Methanol-d ₄ , 500 MHz): δ [ppm] = 8.08 (s, 1H), 7.58-7.52 (m, 3H), 7.26-7.20 ( m, 2H), 6.96 (s, 1H), 4.69 (d, J=12.4 Hz, 2H), 4.60 (s, 2H), 3.95-3.75 (m, 2H ), 3.70 (s, 3H), 3.39-3.24 (m, 2H), 1.44-1.26 (m, 9H).

- 98 039916

HPLC-MS (method A): [m/z]: 496.05 [M+H]+.

tert-Butyl N-(1H-1,3-benzodiazol-2-ylmethyl)-N-(2-{4-[(1,3-oxazol-2-ylmethyl)carbamoyl]-1,3thiazol-2-yl} ethyl)carbamate (56)

Similar to general procedure 6, 2-{2-[(1H-1,3-benzodiazol-2-ylmethyl)[(tert-butoxy)carbonyl]amino]ethyl}-1,3-thiazole-4-carboxylic acid (8) (150 mg, 0.298 mmol, 80% pure), 1,3-oxazol-2ylmethanamine dihydrochloride (102 mg, 0.596 mmol), DIPEA (312 µl, 1.79 mmol) and HATU (227 mg, 0.596 mmol) in DMF ( 3 ml) gave the title compound (94 mg, 63%) as a brownish oil after purification by flash column chromatography (kp-NH, eluting with a 0-100% EtOAc/heptane gradient).

HPLC-MS (method A): [m/z]: 483.05 [M+H] ⁺ .

tert-Butyl N-(1H-1,3-benzodiazol-2-ylmethyl)-N-[2-(4-{[(1-methyl-1 H-pyrazol-3yl)methyl]carbamoyl}-1,3- thiazol-2-yl)ethyl]carbamate (57)

Similar to general procedure 6, 2-{2-[(1H-1,3-benzodiazol-2-ylmethyl)[(tert-butoxy)carbonyl]amino]ethyl}-1,3-thiazole-4-carboxylic acid (8) (150 mg, 0.298 mmol, 80% pure), 1-(1-methyl-1Hpyrazol-3-yl)methanamine (50 mg, 0.45 mmol), DIPEA (156 µl, 0.894 mmol) and HATU (227 mg, 0.596 mmol) in DMF (3 mL) gave the title compound (53 mg, 34%) as a brownish oil after purification by flash column chromatography (kp-NH, eluting with a 0-100% EtOAc/heptane gradient).

HPLC-MS (Method A): [m/z]: 496.45 [M+H] ⁺ .

tert-Butyl N-(1H-1,3-benzodiazol-2-ylmethyl)-N-(2-{4-[(pyridazin-3-ylmethyl)carbamoyl]-1,3thiazol-2-yl}ethyl)carbamate ( 58)

Similar to general procedure 6, 2-{2-[(1H-1,3-benzodiazol-2-ylmethyl)[(tert-butoxy)carbonyl]amino]ethyl}-1,3-thiazole-4-carboxylic acid (8) (100 mg, 0.23 mmol, 92% pure), 1(pyridazin-3-yl)methanamine (37 mg, 0.34 mmol), DIPEA (119 µl, 0.69 mmol) and HATU (130 mg, 0.34 mmol) in DMF (2 mL) gave the title compound (101 mg, 88%) as a pale yellow oil after purification by flash column chromatography (kp-NH, eluting with a gradient of 50-100% EtOAc/heptane, then 0-15% MeOH/EtOAc).

HPLC-MS (Method A): [m/z]: 494.1 [M+H] ⁺ .

tert-Butyl N-(1H-1,3-benzodiazol-2-ylmethyl)-N-[2-(4-{[(1-methyl-1H-pyrazol-5-yl)methyl]carbamoyl}-1,3 -thiazol-2-yl)ethyl]carbamate (59)

n

Similar to general procedure 6, 2-{2-[(1H-1,3-benzodiazol-2-ylmethyl)[(tert-butoxy)carbonyl]amino]ethyl}-1,3-thiazole-4-carboxylic acid (8) (100 mg, 0.23 mmol, 92% pure), 1(1-methyl-1H-pyrazol-5-yl)methanamine (38 mg, 0.34 mmol), DIPEA (119 µl, 0.69 mmol), and HATU ( 130 mg, 0.34 mmol) in DMF (2 mL) gave the title compound (51 mg, 45%) as a pale yellow oil after purification by flash column chromatography (kp-NH, eluting with a gradient of 50 to 100% EtOAc /heptane, then 0-20% MeOH/EtOAc).

HPLC-MS (method A): [m/z]: 496.3 [M+H] ⁺ .

tert-Butyl N-(1H-1,3-benzodiazol-2-ylmethyl)-N-[2-(4-{[(3-fluoropyridin-4-yl)methyl]carbamoyl}1,3-thiazol-2- yl)ethyl]carbamate (60)

- 99 039916

Similar to general procedure 6, 2-{2-[(1H-1,3-benzodiazol-2-ylmethyl)[(tert-butoxy)carbonyl]amino]ethyl}-1,3-thiazole-4-carboxylic acid (8) (100 mg, 0.23 mmol, 92% pure), 1(3-fluoropyridin-4-yl)methanamine (43 mg, 0.34 mmol), DIPEA (119 μl, 0.69 mmol) and HATU (130 mg, 0 .34 mmol) in DMF (3 ml) gave the title compound (137 mg, 83%, 71% pure) as a yellow oil after purification by flash column chromatography (kp-NH, eluting with a gradient of 50-100% EtOAc/heptane ).

¹ H NMR (CDCl ₃ , 500 MHz): δ [ppm] = 10.10 (s, 1H), 8.43 (d, J=6.0 Hz, 1H), 8.37 (dd, J=9.8, 4.9 Hz, 2H), 7.96 (s, 1H), 7.81-7.68 (m, 2H), 7.40 (d, J=8.6 Hz, 1H ), 7.35-7.27 (m, 1H), 7.25-7.22 (m, 1H), 4.68 (d, J=6.0 Hz,

2H), 4.62 (s, 2H), 3.78 (t, J=6.5 Hz, 2H), 3.27-3.23 (m, 2H), 1.37 (s, 9H).

HPLC-MS (method A): [m/z]: 511.15 [M+H]+.

tert-Butyl N(1H-1,3-benzodiazol-2-ylmethyl)-^[2-(4-{[(3-methylpyridin-4-yl)methyl]carbamoyl}1,3-thiazol-2-yl) ethyl]carbamate (61)

Similar to the general procedure

6, 2-{2-[(1H-1,3-benzodiazol-2-ylmethyl) [(tert-butoxy)carbonyl]amino]ethyl}-1,3-thiazole-4-carboxylic acid (8) (700 mg, 1.65 mmol, 95% pure), (3-methylpyridin-4-yl)methanamine dihydrochloride (387 mg, 1.98 mmol), DIPEA (863 μl, 4.9 mmol) and HATU (1260 mg, 3.3 mmol) in DMF (10 mL) gave the title compound (363 mg, 43%) as a yellow oil after purification by flash column chromatography (kp-NH, eluting with a 20-100% EtOAc/heptane gradient).

HPLC-MS (method A): [m/z]: 507.1 [M+H] ⁺ .

tert-Butyl N-(1H-1,3-benzodiazol-2-ylmethyl)-N-[2-(4- {[(1-methyl-1 H-pyrazol-4yl)methyl]carbamoyl}-1,3- thiazol-2-yl)ethyl]carbamate (62)

Similar to general procedure 6, 2-{2-[(1H-1,3-benzodiazol-2-ylmethyl)[(tert-butoxy)carbonyl]amino]ethyl}-1,3-thiazole-4-carboxylic acid (8) (100 mg, 0.25 mmol), (1-methyl-1Hpyrazol-4-yl)methanamine (41 mg, 0.37 mmol), DIPEA (130 µl, 0.75 mmol) and HATU (142 mg, 0.37 mmol ) in DMF (2 mL) gave the title compound (125 mg, quant.) as a yellow oil after purification by flash column chromatography (eluting with a 0-20% MeOH/EtOAc gradient).

HPLC-MS (method A): [m/z]: 496.1 [M+H] ⁺ .

tert-Butyl N-(1H-1,3-benzodiazol-2-ylmethyl)-N-[2-(4-{[(6-methylpyridazin-3 yl)methyl]carbamoyl}-1,3-thiazol-2- yl)ethyl] carbamate (63)

Similar to general procedure 6, 2-{2-[(1H-1,3-benzodiazol-2-ylmethyl)[(tert-butoxy)carbonyl]amino]ethyl}-1,3-thiazole-4-carboxylic acid (8) (100 mg, 0.23 mmol, 92% pure), (6-methylpyridazin-3-yl)methanamine (42 mg, 0.34 mmol), DIPEA (119 µl, 0.69 mmol) and HATU (130 mg, 0.34 mmol) 34 mmol) in DMF (3 mL) gave the crude title compound (99 mg, 67%, 79% pure) after purification by flash column chromatography (kp-NH, eluting with a 70-100% EtOAc/heptane gradient).

1H NMR (CDCl ₃ , 500 MHz): δ [ppm] = 10.50 (s, 1H), 8.28 (s, 1H), 7.92 (s, 1H), 7.72 (s , 1H), 7.40-7.29 (m, 3H), 7.23 (dd, J=6.4, 2.8 Hz, 2H), 4.68 (s, 2H), 3.82 ( s, 2H), 3.24 (s, 2H), 2.73 (s, 2H), 2.71 (s, 3H), 1.36 (s, 9H).

HPLC-MS (Method A): [m/z]: 508.10 [M+H] ⁺ .

tert-Butyl N-(1H-1,3-benzodiazol-2-ylmethyl)-N-(2-{4-[(1H-imidazol-2-ylmethyl)carbamoyl]-1,3thiazol-2-yl}ethyl) carbamate (64)

- 100 039916

n

Similar to general procedure 6, 2-{2-[(III-1,3-benzodiazol-2-ylmethyl)[(tert-butoxy)carbonyl]amino]ethyl}-1,3-thiazole-4-carboxylic acid (8) (100 mg, 0.25 mmol), 1-(1Himidazol-2-yl)methanamine (42 mg, 0.25 mmol), DIPEA (164 µl, 0.99 mmol) and HATU (188 mg, 0.50 mmol) in DMF (2 mL) gave the title compound (65 mg, 54%) as a yellow oil after purification by flash column chromatography (kp-NH, eluting with a 0-5% MeOH/DCM gradient).

HPLC-MS (method A): [m/z]: 482.25 [M+H]+.

tert-Butyl N-(1H-1,3-benzodiazol-2-ylmethyl)-N-{2-[4-({[2-(morpholin-4-yl)pyridin-4yl]methyl}carbamoyl)-1, 3-thiazol-2-yl]ethyl}carbamate (65)

n

Similar to general procedure 6, 2-{2-[(1H-1,3-benzodiazol-2-ylmethyl)[(tert-butoxy)carbonyl]amino]ethyl}-1,3-thiazole-4-carboxylic acid (8) (100 mg, 0.25 mmol), [3-(morpholin4-yl)pyridin-4-yl]methanamine (48 mg, 0.25 mmol), DIPEA (164 μl, 0.99 mmol) and HATU (189 mg, 0 .50 mmol) in DMF (2 mL) gave the title compound (112 mg, 78%) as a yellow solid after purification by flash column chromatography (kp-NH, eluting with a 50-100% EtOAc/heptane gradient) .

HPLC-MS (method A): [m/z]: 578.10 [M+H] ⁺ .

tert-Butyl N-(1H-1,3-benzodiazol-2-ylmethyl)-N-[2-(4-{[(5-methylpyridin-2-yl)methyl]carbamoyl}1,3-thiazol-2- yl)ethyl]carbamate (66)

Similar to general procedure 6, 2-{2-[(1H-1,3-benzodiazol-2-ylmethyl)[(tert-butoxy)carbonyl]amino]ethyl}-1,3-thiazole-4-carboxylic acid (8) (100 mg, 0.20 mmol, 80% purity), 1(5-methylpyridin-2-yl)methanamine (29 mg, 0.24 mmol), DIPEA (104 µl, 0.60 mmol) and HATU (15 lmg, 0 .40 mmol) in DMF (2 mL) gave the title compound (48 mg, 47%) as a colorless oil after purification by flash column chromatography (kp-NH, eluting with a 0-100% EtOAc/heptane gradient).

HPLC-MS (method A): [m/z]: 507.1 [M+H] ⁺ .

tert-Butyl N-(1H-1,3-benzodiazol-2-ylmethyl)-N-{2-[4-({[6-(dimethylamino)pyridin-3yl]methyl}carbamoyl)-1,3-thiazol- 2-yl]ethyl}carbamate (67)

Similar to general procedure 6, 2-{2-[(1H-1,3-benzodiazol-2-ylmethyl)[(tert-butoxy)carbonyl]amino]ethyl}-1,3-thiazole-4-carboxylic acid (8) (100 mg, 0.20 mmol, 80% purity), 5(aminomethyl)-N,N-dimethylpyridin-2-amine (30 mg, 0.20 mmol), DIPEA (104 µl, 0.60 mmol) and HATU (151 mg, 0.40 mmol) in DMF (2 mL) gave the title compound (36 mg, 34%) as a colorless oil after purification by flash column chromatography (kp-NH, eluting with a gradient of 0-100% EtOAc/ heptane).

HPLC-MS (method A): [m/z]: 536.35 [M+H] ⁺ .

tert-Butyl N-(1H-1,3-benzodiazol-2-ylmethyl)-N-[2-(4-{[(2-methylpyridin-4-yl)methyl]carbamoyl}1,3-thiazol-2- yl)ethyl]carbamate (68)

Similar to general procedure 6, 2-{2-[(1H-1,3-benzodiazol-2-ylmethyl)[(tert- 101 039916 butoxy)carbonyl]amino]ethyl}-1,3-thiazol-4-carboxylic acid ( 8) (100 mg, 0.20 mmol, 80% purity), 1(2-methylpyridin-4-yl)methanamine (36 mg, 0.30 mmol), DIPEA (104 µl, 0.60 mmol) and HATU ( 151 mg

0.40 mmol) in DMF (2 mL) gave the title compound (36 mg, 36%) as a colorless oil after purification by flash column chromatography (kp-NH, eluting with a 0-100% gradient).

EtOAc/heptane).

HPLC-MS (method A): [m/z]: 507.3 [M+H]+.

tert-Butyl N-(1H-1,3-benzodiazol-2-ylmethyl)-N-[2-(4-{[(1,5-dimethyl-1 H-pyrazol-4yl)methyl]carbamoyl}-1, 3-thiazol-2-yl)ethyl]carbamate (69)

Similar to general procedure 6, 2-{2-[(1H-1,3-benzodiazol-2-ylmethyl)[(tert-butoxy)carbonyl]amino]ethyl}-1,3-thiazole-4-carboxylic acid (8) (100 mg, 0.20 mmol, 80% pure), 1(1,5-dimethyl-1H-pyrazol-4-yl)methanamine (37 mg, 0.30 mmol), DIPEA (104 µl, 0.60 mmol) and HATU (151 mg, 0.40 mmol) in DMF (2 mL) gave the title compound (74 mg, 73%) as a colorless oil after purification by flash column chromatography (kp-NH, eluting with a gradient of 0100% EtOAc /heptane).

HPLC-MS (method A): [m/z]: 510.15 [M+H] ⁺ .

tert-Butyl N-(1H-1,3-benzodiazol-2-ylmethyl)-N-{2-[4-({[3-chloro-5-(trifluoromethyl)pyridine-2yl]methyl}carbamoyl)-1, 3-thiazol-2-yl]ethyl} carbamate (70)

n

Similar to general procedure 6, 2-{2-[(1H-1,3-benzodiazol-2-ylmethyl)[(tert-butoxy)carbonyl]amino]ethyl}-1,3-thiazole-4-carboxylic acid (8) (100 mg, 0.236 mmol), 1-[3-chloro-5(trifluoromethyl)pyridin-2-yl]methanamine hydrochloride (87 mg, 0.354 mmol), DIPEA (0.21 ml, 1.18 mmol), and HATU (135 mg , 0.354 mmol) in DMF (3 mL) gave the title compound (216 mg, 69%, 45% pure) as a yellow oil after purification by flash column chromatography (KP-NH, eluting with a gradient of 20-100% EtOAc /heptane). The resulting title compound was used in the next step without further purification.

HPLC-MS (method A): [m/z]: 595.1 [M+H] ⁺ .

tert-Butyl N-(1H-1,3-benzodiazol-2-ylmethyl)-N-[2-(4-{[(3-chloro-5-fluoropyridine-2yl)methyl]carbamoyl}-1,3-thiazole -2-yl)ethyl]carbamate (71)

Similar to general procedure 6, 2-{2-[(1H-1,3-benzodiazol-2-ylmethyl)[(tert-butoxy)carbonyl]amino]ethyl}-1,3-thiazole-4-carboxylic acid (8) (100 mg, 0.236 mmol), (3-chloro-5fluoropyridin-2-yl)methanamine hydrochloride (70 mg, 0.354 mmol), DIPEA (0.21 ml, 1.18 mmol) and HATU (135 mg, 0.354 mmol) in DMF (3 ml) gave the title compound (157 mg, 76%, 62% pure) as a yellow oil after purification by flash column chromatography (KP-NH, eluting with a 20-100% EtOAc/heptane gradient). The resulting title compound was used in the next step without further purification.

HPLC-MS (method A): [m/z]: 545.15 [M+H] ⁺ .

tert-Butyl N-(1H-1,3-benzodiazol-2-ylmethyl)-N-[2-(4-{[(2-fluoropyridin-3-yl)methyl]carbamoyl}1,3-thiazol-2- yl)ethyl]carbamate (72)

Similar to general procedure 6, 2-{2-[(1H-1,3-benzodiazol-2-ylmethyl)[(tert-butoxy)carbonyl]amino]ethyl}-1,3-thiazole-4-carboxylic acid (8) (100 mg, 0.236 mmol, 95% pure), 1-(2-fluoropyridin-3-yl)methanamine (47.01 mg, 0.373 mmol), DIPEA (0.13 ml, 0.745 mmol) and HATU (141.7

- 102039916 mg, 0.373 mmol) in DMF (2 ml) gave the title compound (0.359 g, quant.) as a brown solid after evaporating the solvent. The resulting title compound was used in the next step without further purification.

HPLC-MS (Method A): [m/z]: 511.10 [M+H]+.

tert-Butyl X-(1H-1,3-benzodiazol-2-ylmethyl)-X-[2-(4-{[(2-methoxypyridin-4yl)methyl]carbamoyl}-1,3-thiazol-2-yl )ethyl] carbamate (73)

n

Similar to general procedure 6, 2-{2-[(1H-1,3-benzodiazol-2-ylmethyl)[(tert-butoxy)carbonyl]amino]ethyl}-1,3-thiazole-4-carboxylic acid (8) (100 mg, 0.236 mmol, 95% pure), 1-(2-methoxypyridin-4-yl)methanamine (49 mg, 0.354 mmol), DIPEA (0.12 ml, 0.708 mmol) and HATU (135 mg, 0.354 mmol) in DMF (3 mL) gave the title compound (104 mg, 81%, 96% pure) as a white solid after purification by flash column chromatography (eluting with a 30-100% EtOAc/heptane gradient).

¹ H NMR (CDCl3, 500 MHz): δ [ppm] = 10.07 (s, 1H), 8.10 (d, J=5.3 Hz, 1H), 7.96 (s, 1H ), 7.73-7.66 (m, 2H), 7.41-7.37 (m, 1H), 7.25-7.22 (m, 2H), 6.85-6.82 (m , 1H), 6.69 (s, 1H), 4.62 (s, 2H), 4.56 (d, J=6.3 Hz, 2H), 3.91 (s, 3H), 3.78 (t, J=6.6 Hz, 2H), 3.24 (t, J=6.2 Hz, 2H), 1.39 (s, 9H).

HPLC-MS (method A): [m/z]: 523.3 [M+H] ⁺ .

tert-Butyl N-(1H-1,3-benzodiazol-2-ylmethyl)-N-[2-(4-{[(4,6-dimethylpyridin-3yl)methyl]carbamoyl}-1,3-thiazol-2 -yl)ethyl]carbamate (74)

Similar to general procedure 6, 2-{2-[(1H-1,3-benzodiazol-2-ylmethyl)[(tert-butoxy)carbonyl]amino]ethyl}-1,3-thiazole-4-carboxylic acid (8) (100 mg, 0.236 mmol, 95% pure), (4,6-dimethylpyridin-3-yl)methanamine dihydrochloride (B1) (93 mg, 0.354 mmol, 80% pure), DIPEA (0.206 ml, 1.18 mmol) and HATU (135 mg, 0.354 mmol) in DMF (3 mL) gave the title compound (77 mg, 63%) as a yellow oil after purification by flash column chromatography (eluting with a 0-15% MeOH/EtOAc gradient).

HPLC-MS (method A): [m/z]: 521.05 [M+H] ⁺ .

tert-Butyl N-(1H-1,3-benzodiazol-2-ylmethyl)-N -[2-(4-{[(4-methylpyridin-2-yl)methyl]carbamoyl} 1,3-thiazol-2- yl)ethyl]carbamate (75)

n

Similar to general procedure 6, 2-{2-[(1H-1,3-benzodiazol-2-ylmethyl)[(tert-butoxy)carbonyl]amino]ethyl}-1,3-thiazole-4-carboxylic acid (8) (100 mg, 0.236 mmol, 95% pure), (4,6-dimethylpyridin-3-yl)methanamine dihydrochloride (93 mg, 0.354 mmol), DIPEA (0.123 ml, 0.708 mmol) and HATU (135 mg, 0.354 mmol) in DMF (2 ml) gave the title compound (90 mg, 72%) as a yellow oil after purification by flash column chromatography (KP-NH, eluting with a gradient of 20-100% EtOAc/heptane followed by 0-20% MeOH/ EtOAc).

HPLC-MS (method A): [m/z]: 507.10 [M+H] ⁺ .

tert-Butyl N-(1H-1,3-benzodiazol-2-ylmethyl)-N-{2-[4-(5,6,7,8-tetrahydro-1,6-naphthyridine-6carbonyl)-1,3 -thiazol-2-yl]ethyl}carbamate (76)

Similar to general procedure 6, 2-{2-[(1H-1,3-benzodiazol-2-ylmethyl)[(tert-butoxy)carbonyl]amino]ethyl}-1,3-thiazole-4-carboxylic acid (8) (100 mg, 0.236 mmol, 95% pure), 5,6,7,8-tetrahydro-1,6-naphthyridine dihydrochloride (62 mg, 0.298 mmol), DIPEA (0.173 ml, 0.994 mmol) and HATU (151 mg, 0.398 mmol ) in DMF (2 mL) gave the title compound (90 mg, 72%) as a colorless oil after purification by flash column chromatography (kp-NH, eluting in

- 103 039916 gradient 0-100% EtOAc/heptane).

HPLC-MS (method A): [m/z]: 519.15 [M+H]+.

tert-Butyl N-(1H-1,3-benzodiazol-2-ylmethyl)-N-[2-(4-{[(3,5-difluoropyridin-2yl)methyl]carbamoyl}-1,3-thiazol-2 -yl)ethyl]carbamate (77)

Similar to general procedure 6, 2-{2-[(1H-1,3-benzodiazol-2-ylmethyl)[(tert-butoxy)carbonyl]amino]ethyl}-1,3-thiazole-4-carboxylic acid (8) (100 mg, 0.236 mmol, 95% pure), (3,5-difluoropyridin-2-yl)methanamine dihydrochloride (65 mg, 0.298 mmol), DIPEA (0.173 ml, 0.994 mmol) and HATU (151 mg, 0.398 mmol) in DMF (3 ml) gave the title compound (112 mg, quant.) as a colorless oil after purification by flash column chromatography (kp-NH, eluting with a 0-100% EtOAc/heptane gradient).

HPLC-MS (method A): [m/z]: 529.10 [M+H] ⁺ .

tert-Butyl N-(1H-1,3-benzodiazol-2-ylmethyl)-N-[2-(4-{[(3-{[(tert-butyldimethylsilyl)oxy]methyl}pyridin-2-yl)methyl ]carbamoyl}-1,3-thiazol-2-yl)ethyl]carbamate (78)

Similar to general procedure 6, 2-{2-[(1H-1,3-benzodiazol-2-ylmethyl)[(tert-butoxy)carbonyl]amino]ethyl}-1,3-thiazole-4-carboxylic acid (8) (15 .94 mg, 0.04 mmol), (3-{[(tert-butyldimethylsilyl)oxy]methyl}pyridin-2-yl)methanamine (C3) (10 mg, 0.04 mmol), DIPEA (0.03 ml, 0 .16 mmol) and HATU (30.13 mg, 0.08 mmol) in DMF (2 mL) gave the title compound (17.5 mg, 34%, 30% pure) as an orange oil after column flash purification. -chromatography (elution with a gradient of 0-100% EtOAc/heptane).

HPLC-MS (method A): [m/z]: 637.15 [M+H] ⁺ .

tert-Butyl 2-{[(2-{2-[(1H-1,3-benzodiazol-2-ylmethyl)[(tert-butoxy)carbonyl]amino]ethyl}-1,3thiazol-4-yl)formamido] methyl} piperidine-1-carboxylate (79)

Similar to general procedure 6, 2-{2-[(1H-1,3-benzodiazol-2-ylmethyl)[(tert-butoxy)carbonyl]amino]ethyl}-1,3-thiazole-4-carboxylic acid (8) (200 mg, 0.5 mmol), tert-butyl 2(aminomethyl)piperidine-1-carboxylate (149 mg, 0.7 mmol), TEA (66.16 µl, 0.5 mmol) and HATU (280 mg, 0.5 mmol) 75 mmol) in DMF (5 ml) at room temperature for 2 h gave the title compound (50 mg, 17%) as an orange oil after purification by flash column chromatography (eluting with a gradient of 0-100% EtOAc/heptane ) followed by basic prep-HPLC.

HPLC-MS (method A): [m/z]: 599.4 [M+H] ⁺ .

2-{2-[(1H-1,3-Benzodiazol-2-ylmethyl)amino]ethyl}-N-(pyridin-2-ylmethyl)-1,3-thiazol-4carboxamide trihydrochloride (illustrative example No. 12) o

Similar to general procedure 2, 4M HC in dioxane (11 mL) and tert-butyl N(1^1,3benzodiazol-2-ylmethyl)-N-(2-{4-[(pyridin-2-ylmethyl)carbamoyl]- 1,3-thiazol-2-yl}ethyl)carbamate (9) (2.2 g, 4.47 mmol) in dioxane (30 mL) at room temperature over 16 h gave the title compound (HC salt) ( 1.7 g, 76%) as a yellow solid after trituration in Et ₂ O (2x30 ml) and then in DCM (2x20 ml) and Et ₂ O (2x30 ml).

¹ H NMR (DMSO-d ₆ , 500 MHz): δ [ppm] = 10.39 (s, 1H), 9.68 (t, J=6.0 Hz, 1H), 8.86- 8.75 (m, 1H), 8.44

- 104 039916 (td, J=7.9, 1.5 Hz, 1H), 8.30 (s, 1H), 7.96-7.84 (m, 2H), 7.76 (dt, J= 6.5, 3.3 Hz, 2H), 7.44 (d.s., J=6.5, 3.4 Hz,

2H), 4.86 (d, J=6.0 Hz, 2H), 4.76 (s, 2H), 3.66 (dt, J=38.8, 7.1 Hz, 4H).

HPLC-MS (method C): [m/z]: 493.4 [M+H]+.

2-{2-[(1H-1,3-Benzodiazol-2-ylmethyl)amino]ethyl}-N(thiophen-2-ylmethyl)-1,3-thiazol-4-carboxamide (illustrative example No. 16)

Similar to general procedure 7, tert-butyl N-(1H-1,3-benzodiazol-2-ylmethyl)-N-(2-{4-[(thiophen-2ylmethyl)carbamoyl]-1,3-thiazol-2-yl }ethyl)carbamate (12) (180 mg, 0.362 mmol) and 50% TFA in DCM (10 mL) at room temperature overnight gave the title compound (51 mg, 34%, 98% pure) in as a colorless oil after purification by preparative HPLC.

1H NMR (CDCl ₃ , 400 MHz): δ [ppm] = 8.05 (s, 1H), 7.59 (m, 3H), 7.24 (m, 2H), 7.21-7 .19 (dd, J=5.1, 1.2 Hz, 1H), 7.01 (m, 1H), 6.94 (m, 1H), 4.78 (d, J=6.0, 2H ), 4.15 (s, 2H), 3.27-3.07 (m, 4H).

HPLC-MS (method J): [m/z]: 398.5 [M+H] ⁺ .

2-{2-[(1H-1,3-Benzodiazol-2-ylmethyl)amino]ethyl}-N-methyl-K-phenyl-1,3-thiazole-4-carboxamide (illustrative example No. 19)

Similar to general procedure 2, tert-butyl N(1H-1,3-benzodiazol-2-ylmethyl)-N(2-{4[methyl(phenyl)carbamoyl]-1,3-thiazol-2-yl}ethyl)carbamate (15) (110.12 mg, 0.224 mmol) and 50% TFA in DCM (10 mL) at room temperature overnight gave the title compound (40 mg, 45%, 85% pure) as a colorless oils after purification by flash column chromatography (gradient elution with 0-10% MeOH in DCM).

1H NMR (CDCl ₃ , 400 MHz): δ [ppm] = 7.69 (br s, 2H), 7.29-7.21 (m, 5H), 7.10 (s, 2H) , 4.28 (s, 2H), 3.60-3.43 (m, 3H), 3.18 (br s, 2H), 3.05 (br s, 2H).

HPLC-MS (method J): [m/z]: 392.5 [M+H] ⁺ .

2-{2-[(1H-1,3-Benzodiazol-2-ylmethyl)amino]ethyl} -N-{[2-(pyrrolidin-1-yl)phenyl]methyl}-1,3thiazol-4-carboxamide ( illustrative example No. 21)

Similar to general procedure 2, tert-butyl N-(1H-1,3-benzodiazol-2-ylmethyl)-N-{2-[4-({[2(pyrrolidin-1-yl)phenyl]methyl}carbamoyl)-1, 3-thiazol-2-yl]ethyl}carbamate (16) (100 mg, 0.178 mmol) and 50% TFA in DCM (8 ml) at room temperature overnight gave the title compound (70 mg, 69 %, 82% purity) as a colorless oil after purification by flash column chromatography (eluting with a gradient of 5-7% MeOH in DCM).

1H NMR (CDCl ₃ , 400 MHz): δ [ppm] = 7.94 (s, 1H), 7.85 (t, J=5.6 Hz, 1H), 7.53-7.47 (m, 2H), 7.24-7.16 (m, 2H), 7.12 (t, J=7.8 Hz, 1H), 6.59 (d, J=7.5 Hz, 1H) , 6.48 (s, 1H), 6.43 (d, J=8.1 Hz, 1H), 4.53 (d, J=5.9 Hz, 2H), 4.14 (s, 2H) , 3.21 (m, 4H), 3.12-3.05 (m, 4H), 2.00-1.90 (m, 4H).

HPLC-MS (method J): [m/z]: 461.6 [M+H] ⁺ .

2-{2-[(1H-1,3-Benzodiazol-2-ylmethyl)amino]ethyl}-N(pyridin-3-ylmethyl)-1,3-thiazol-4carboxamide trihydrochloride (illustrative example No. 35)

Similar to general procedure 2, tert-butyl N-(1H-1,3-benzodiazol-2-ylmethyl)-N-(2-{4-[(pyridin-3ylmethyl)carbamoyl]-1,3-thiazol-2-yl }ethyl)carbamate (22) (184 mg, 0.374 mmol) and 4M HC solution in dioxane (15 ml) at room temperature for 18 hours gave the title compound (80 mg, 53%) as trihydrochloride salt as white solid after precipitation with diethyl ether.

¹ H NMR (DMSO-H6, 400 MHz): δ [ppm] = 10.28 (brs, 3H), 9.64 (t, J=6.2 Hz, 1H), 8.90 (s, 1H), 8.81 (d, J=5.4Hz, 1h), 8.55 (d, J=8.1Hz, 1H), 8.25 (s, 1H), 8.00 ( dd, J=8.0, 5.7 Hz, 1H), 7.73 (m, 2H), 7.48-7.34 (m,

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2H), 4.71 (s, 2H), 4.65 (d, J=6.2 Hz, 2H), 3.66 (t, J=6.6 Hz, 2H), 3.57 (t, J=6.3 Hz, 2H).

HPLC-MS (method J): [m/z]: 493.3 [M+H]+.

2-{2-[(1H-1,3-Benzodiazol-2-ylmethyl)amino]ethyl}-N-(pyridin-4-ylmethyl)-1,3-thiazol-4carboxamide trihydrochloride (illustrative example No. 36)

Similar to general procedure 2, tert-butyl N-(1H-1,3-benzodiazol-2-ylmethyl)-N-(2-{4-[(pyridin-4ylmethyl)carbamoyl]-1,3-thiazol-2-yl }ethyl)carbamate (23) (145.8 mg, 0.296 mmol) and a 4M solution of HC in dioxane (15 ml) at room temperature for 18 hours gave the title compound (70 mg, 47%) as the trihydrochloride salt in as a white solid after precipitation with diethyl ether.

¹ H NMR (DMSO, 400 MHz): δ [ppm] = 10.40 (brs, 3H), 9.69 (t, J=6.2 Hz, 1H), 8.83 (d , J=6.7 Hz, 2H), 8.28 (s, 1H), 7.98 (d, J=6.6 Hz, 2H), 7.81-7.69 (m, 2H), 7 .49-7.37 (m, 2H), 4.73 (d, J=5.1 Hz, 4H), 3.68 (t, J=6.5 Hz, 2H), 3.60 (t, J=6.4 Hz, 2H).

HPLC-MS (method J): [m/z]: 493.3 [M+H] ⁺ .

2-{2-[(1H-1,3-Benzodiazol-2-ylmethyl)amino]ethyl}-N-{[3-(trifluoromethyl)pyridin-2-yl]methyl}-1,3thiazol-4-carboxamide ( illustrative example No. 37)

Similar to general procedure 2, tert-butyl N-(1H-1,3-benzodiazol-2-ylmethyl)-N-{2-[4-({[3(trifluoromethyl)pyridin-2-yl]methyl}carbamoyl)- 1,3-thiazol-2-yl]ethyl}carbamate (24) (86.33 mg, 0.154 mmol) and 4M HC in dioxane (10 ml) at room temperature for 18 h gave the title compound (25 mg , 35%) as a yellow oil after purification by flash column chromatography (gradient elution with 10-15% MeOH in DCM).

1H NMR (MeOD, 400 MHz): δ [ppm] = 8.62 (d, J=4.8 Hz, 1H), 8.13-8.09 (m, 1H), 8.08 ( s, 1H), 7.52-7.47 (m, 2H), 7.47-7.41 (m, 1H), 7.22-7.14 (m, 2H), 4.86 (s, 2H), 4.09 (s, 2H), 3.27 (t, J=6.6 Hz, 2H), 3.13 (t, J=6.6 Hz, 2H).

HPLC-MS (method J): [m/z]: 461.6 [M+H] ⁺ .

2-{2-[(1H-1,3-Benzodiazol-2-ylmethyl)amino]ethyl}-N-(5,6,7,8-tetrahydroquinolin-8-ylmethyl)-1,3thiazol-4-carboxamide ( illustrative example No. 38)

Similar to general procedure 2, tert-butyl N-(1H-1,3-benzodiazol-2-ylmethyl)-N-(2-{4-[(5,6,7,8tetrahydroquinolin-8-ylmethyl)carbamoyl]-1 ,3-thiazol-2-yl}ethyl)carbamate (25) (82.03 mg, 0.154 mmol) and 4M HC in dioxane (10 ml) at room temperature for 18 hours gave the title compound (35 mg, 52%) as a yellow oil after purification by flash column chromatography (gradient elution with 10-15% MeOH in DCM).

1H NMR (MeOD, 400 MHz): δ [ppm] = 8.27 (d, J=3.6 Hz, 1H), 8.08 (s, 1H), 7.58 (d, J= 7.6 Hz, 1H), 7.49 (m, 2H), 7.27-7.15 (m, 3H), 5.22-5.12 (m, 1H), 4.05 (s, 2H ), 3.21 (t, J=6.7 Hz, 2H), 3.05 (t, J=6.4 Hz, 2H), 2.962.77 (m, 2H), 2.34-2.20 (m, 1H), 2.05-1.79 (m, 3H).

HPLC-MS (method J): [m/z]: 433.6 [M+H]+.

2-{2-[(1H-1,3-Benzodiazol-2-ylmethyl)amino]ethyl} -N- {5H,6H,7H-cyclopenta[b]pyridin-7-yl} -1,3thiazol-4- carboxamide trihydrochloride (illustrative example No. 39)

Similar to general procedure 2, a 4M solution of HC in dioxane (6.36 ml) was added to a solution of tert-butyl N-(1H-1,3-benzodiazol-2-ylmethyl)-N-{2-[4-({5H, 6H,7H-cyclopenta[b]pyridin-7-yl}carbamoyl)-1,3thiazol 2-yl]ethyl}carbamate (26) (1.32 g, 2.55 mmol) in dioxane (30 ml) and stirred at room temperature for 48 hours to give the title compound (1.03 g, 76%) after crystallization from DCM/MeOH.

1H NMR (MeOD, 500 MHz): δ [ppm] = 8.56 (d, J=5.8 Hz, 1H), 8.52-8.48 (m, 1H), 8.31 ( s, 1H), 7.92 (dd, J=7.7, 6.0 Hz, 1H), 7.86 (dt, J=6.7, 3.3 Hz, 2H), 7.66 (dt , J=6.3, 3.3 Hz, 2H), 5.95 (t, J=8.8 Hz, 1H), 5.01 (s, 2H), 3.87 (t, J=6, 2 Hz, 2H), 3.71-3.63 (m, 2H), 3.42-3.35 (m, 1H), 3.21 (dt, J=17.1, 8.7 Hz, 1H ), 2.86-2.77 (m, 1H), 2.51 (d.sq., J=12.9, 9.2 Hz, 1H).

HPLC-MS (Method C): [m/z]: 419.05 [M+H]+.

2-{2-[(1H-1,3-Benzodiazol-2-ylmethyl)amino]ethyl}-N[(3-fluoropyridin-2-yl)methyl]-1,3-thiazole-4carboxamide trihydrochloride (Illustrative Example No. 40)

Similar to general procedure 2, a 4M solution of HC in dioxane (16.57 mL, 66.26 mmol) was added to a solution of tert-butyl N-(1H-1,3-benzodiazol-2-ylmethyl)-N-[2-(4 -{[(3-fluoropyridin-2-yl)methyl]carbamoyl}1,3-thiazol-2-yl)ethyl]carbamate (27) (4.13 g, 6.63 mmol) in dioxane (40 ml) and stirred at room temperature for 16 h to give the title compound (3.04 g, 88%) as a white solid after precipitation from diethyl ether (100 mL).

1H NMR (MeOD, 500 MHz): δ [ppm] = 8.47 (d, J=5.1 Hz, 1H), 8.28 (s, 1H), 8.11 (s, 1H) , 7.84 (dd, J=6.1, 3.1 Hz, 2H), 7.76 (s, 1H), 7.62 (dd, J=6.1, 3.0 Hz, 2H), 4.99 (s, 2H), 4.95 (s, 2H), 3.84 (t, J=6.2 Hz, 2H), 3.65 (t, J=6.2 Hz, 2H).

HPLC-MS (Method D): [m/z]: 411.1 [M+H] ⁺ .

2-{2-[(1H-1,3-Benzodiazol-2-ylmethyl)amino]ethyl}-N-[(6-methylpyridin-2-yl)methyl]-1,3-thiazole-4carboxamide (illustrative example no. 42)

Similar to general procedure 2, tert-butyl N-(1H-1,3-benzodiazol-2-ylmethyl)-N-[2-(4-{[(6methylpyridin-2-yl)methyl]carbamoyl}-1,3- thiazol-2-yl)ethyl]carbamate (29) (79.5 mg, 0.157 mmol) and 4M HC in dioxane (15 ml) at room temperature overnight gave the title compound (25.5 mg, 36% ) as a pale yellow oil after purification by flash column chromatography (gradient elution with 10-15% MeOH in DCM).

1H NMR (MeOD, 400 MHz): δ [ppm] = 8.09 (s, 1H), 7.63 (t, J=7.7 Hz, 1H), 7.50 (m, 2H) , 7.23-7.17 (m, 3H), 7.15 (m, 2H), 4.63 (s, 2H), 4.08 (s, 2H), 3.26 (t, J=6 .7 Hz, 2H), 3.11 (t, J=6.6 Hz, 2H), 2.50 (s, 3H).

HPLC-MS (method J): [m/z]: 405.5 [M+H] ⁺ .

2-{2-[(1H-1,3-Benzodiazol-2-ylmethyl)amino]ethyl}-N-[(5-fluoropyridin-2-yl)methyl]-1,3-thiazole-4carboxamide (illustrative example no. 43)

Similar to general procedure 2, tert-butyl N-(1H-1,3-benzodiazol-2-ylmethyl)-N-[2-(4-{[(5fluoropyridin-2-yl)methyl]carbamoyl}-1,3- thiazol-2-yl)ethyl]carbamate (30) (80.2 mg, 0.157 mmol) and 4M HC in dioxane (15 ml) at room temperature overnight gave the title compound (12.5 mg, 17% ) as a pale yellow oil after purification by flash column chromatography (gradient elution with 10-15% MeOH in DCM).

1H NMR (MeOD, 400 MHz): δ [ppm] = 8.36 (d, J=2.6 Hz, 1H), 8.08 (s, 1H), 7.59-7.46 ( m, 3H), 7.42 (m, 1H), 7.20 (m, 2H), 4.66 (s, 2H), 4.08 (s, 2H), 3.26 (t, J=6 .5 Hz, 2H), 3.11 (t, J=6.5 Hz, 2H).

HPLC-MS (method J): [m/z]: 411.5 [M+H] ⁺ .

2-{2-[(1H-1,3-Benzodiazol-2-ylmethyl)amino]ethyl}-N-(pyrimidin-4-ylmethyl)-1,3-thiazol-4carboxamide (illustrative example No. 44)

Similar to the general procedure

2, tert-butyl N-(1H-1,3-benzodiazol-2-ylmethyl)-N-(2-{4

[(pyrimidin-4-ylmethyl)carbamoyl]-1,3-thiazol-2-yl}ethyl)carbamate (31) (75 mg, 0.152 mmol) and 4M solution of 107 039916 HC in dioxane (2 ml) at room temperature temperature over 4 hours gave the title compound (22 mg, 37%) as a yellow solid after purification by flash column chromatography (gradient elution with 8% MeOH in DCM).

1H NMR (MeOD, 400 MHz): δ [ppm] = 9.05 (d, J=1.1 Hz, 1H), 8.67 (d, J=5.3 Hz, 1H), 8 .12 (s, 1H), 7.51 (dd, J=6.0, 3.2 Hz, 2H), 7.46 (d, J=5.3 Hz, 1H), 7.26-7, 18 (m, 2H), 4.68 (s, 2H), 4.10 (s, 2H), 3.28 (t, J=6.5 Hz, 2H), 3.14 (t, J=6 .6 Hz, 2H).

HPLC-MS (method J): [m/z]: 394.4 [M+H] ⁺ .

2-{2-[(1H-1,3-Benzodiazol-2-ylmethyl)amino]ethyl}-N-[(5-methoxypyridin-2-yl)methyl]-1,3-thiazole-4carboxamide (illustrative example no. 45)

Similar to general procedure 2, tert-butyl N-(1H-1,3-benzodiazol-2-ylmethyl)-N-[2-(4-{[(5methoxypyridin-2-yl)methyl]carbamoyl}-1,3- thiazol-2-yl)ethyl]carbamate (32) (80 mg, 0.153 mmol) and 4M HC in dioxane (2 mL) at room temperature for 4 h gave the title compound (53 mg, 82%) as yellow solid after purification by flash column chromatography (gradient elution with 8% MeOH in DCM).

1H NMR (MeOD, 400 MHz): δ [ppm] = 8.17-8.13 (m, 1H), 8.08 (s, 1H), 7.54-7.47 (m, 2H ), 7.35-7.31 (m, 2H), 7.25-7.18 (m, 2H), 4.61 (s, 2H), 4.07 (d, J=8.0 Hz, 2H), 3.84 (s, 3H), 3.26 (t, J=6.6 Hz, 2H), 3.11 (t, J=6.6 Hz, 2H).

HPLC-MS (method J): [m/z]: 423.4 [M+H]+.

2-{2-[(1H-1,3-Benzodiazol-2-ylmethyl)amino]ethyl}-N-(pyrazin-2-ylmethyl)-1,3-thiazol-4carboxamide (illustrative example No. 46)

Similar to general procedure 2, tert-butyl N-(1H-1,3-benzodiazol-2-ylmethyl)-N-(2-{4-[(pyrazin-2ylmethyl)carbamoyl]-1,3-thiazol-2-yl }ethyl)carbamate (33) (70 mg, 0.142 mmol) and 4M HC solution in dioxane (2 ml) at room temperature for 4 hours gave the title compound (30 mg, 53.7%) as a brown solid after purification by flash column chromatography (gradient elution with 8% MeOH in DCM).

1H NMR (MeOD, 400 MHz): δ [ppm] = 8.63 (s, 1H), 8.56-8.51 (m, 1H), 8.48 (d, J=2.6 Hz, 1H), 8.12 (s, 1H), 7.52 (dd, J=6.0, 3.2 Hz, 2H), 7.22 (dd, J=6.1, 3.1 Hz , 2H), 4.74 (s, 2H), 4.13 (s, 2H), 3.28 (t, J=6.5Hz, 2H), 3.17 (t, J=6.5Hz, 2H).

HPLC-MS (method I): [m/z]: 394.4 [M+H] ⁺ .

2-{2-[(1H-1,3-Benzodiazol-2-ylmethyl)amino]ethyl}-N-[(6-oxo-1,6-dihydropyridin-2-yl)methyl]-1,3thiazol-4 -carboxamide (illustrative example No. 47)

Similar to general procedure 2, tert-butyl N-(1H-1,3-benzodiazol-2-ylmethyl)-N-[2-(4-{[(6-oxo-1,6dihydropyridin-2-yl)methyl]carbamoyl }-1,3-thiazol-2-yl)ethyl]carbamate (34) (70 mg, 0.138 mmol) and 4M solution of HC in dioxane HC (2 ml) at room temperature for 4 hours gave the title compound (30 mg, 53%) as a yellow solid after purification by flash column chromatography (gradient elution with 8% MeOH in DCM).

1H NMR (MeOD, 400 MHz): δ [ppm] = 8.12 (s, 1H), 7.51 (m, 3H), 7.22 (dd, J=6.0, 3.2 Hz, 2H), 6.42 (d, J=9.1 Hz, 1H), 6.30 (d, J=6.9 Hz, 1H), 4.44 (s, 2H), 4.09 ( s, 2H), 3.27 (t, J=6.7 Hz, 2H), 3.12 (t, J=6.7 Hz, 2H).

HPLC-MS (method J): [m/z]: 409.4 [M+H] ⁺ .

2-{2-[(1H-1,3-Benzodiazol-2-ylmethyl)amino]ethyl}-N-[(6-cyanopyridin-3-yl)methyl]-1,3-thiazole-4carboxamide (Illustrative Example No. 56) and 5-{[(2-{2-[(1H-1,3-Benzodiazol-2-ylmethyl)amino]ethyl}1,3-thiazol-4-yl)formamido]methyl}pyridine-2-carboxamide (illustrative example #54)

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Similar to general procedure 2, tert-butyl N-(1H-1,3-benzodiazol-2-ylmethyl)-N-[2-(4-{[(6carbamoylpyridin-3-yl)methyl]carbamoyl}-1,3 -thiazol-2-yl)ethyl]carbamate (35) and tert-butyl N-(1H-1,3benzodiazol-2-ylmethyl)-N -[2-(4-{[(6-cyanopyridin-3-yl) methyl]carbamoyl }-1,3-thiazol-2yl)ethyl]carbamate (36) (80 mg, 0.155 mmol) and a 4M solution of HC in dioxane (3 ml) at room temperature for 18 hours gave two products. Formamide (illustrative example No. 54) (16 mg, 23%) was isolated after flash column chromatography (elution with a DCM/MeOH gradient, 9:1). The crude nitrile (Illustrative Example No. 56) was also isolated and further purified by basic prep-HPLC to give the title product as a brown solid (24 mg, 37%).

Formamide: 1H NMR (Methanol-b ₄ , 400 MHz): δ [ppm] = 8.49 (t, J=10.2 Hz, 1H), 7.99 (s, 1H), 7.94 (t, J=10.0 Hz, 1H), 7.78 (dt, J=8.1, 4.0 Hz, 1H), 7.46-7.34 (m, 2H), 7.11 ( dd, J=6.0, 3.1 Hz, 2H), 4.56 (d, J=14.2 Hz, 2H), 4.02 (d, J=8.0 Hz, 2H), 3, 19 (m, 4H), 3.06 (t, J=6.5 Hz, 2H).

HPLC-MS (Method I): [m/z]: 436.5 [M+H]+.

Nitrile: ¹ H NMR (DMSO-66, 500 MHz): δ [ppm] = 12.18 (s, 1H), 9.08 (t, J=6.2 Hz, 1H), 8.70 (s, 1H), 8.14 (s, 1H), 7.97 (d, J=8.0 Hz, 1H), 7.92 (dd, J=8.1, 2.0 Hz, 1H) , 7.48 (d, J=29.4 Hz, 2H), 7.12 (d, J=4.7 Hz, 2H), 4.54 (d, J=6.2 Hz, 2H), 4 .10 (d, J=5.2 Hz, 1H), 3.96 (s, 2H), 3.18-3.15 (m, 2H), 2.98 (d, J=6.6 Hz, 2H).

HPLC-MS (method I): [m/z]: 418.2 [M+H] ⁺ .

2-{2-[(1H-1,3-Benzodiazol-2-ylmethyl)amino]ethyl}-N-[(3,5-dimethylpyridin-2-yl)methyl]-1,3thiazole-4-carboxamide (illustrative example no. 57)

Similar to general procedure 2, tert-butyl N-(1H-1,3-benzodiazol-2-ylmethyl)-N-[2-(4-{[(3,5dimethylpyridin-2-yl)methyl]carbamoyl}-1, 3-thiazol-2-yl)ethyl]carbamate (37) (198 mg, 0.36 mmol) and 12M solution of HC (0.307 ml, 8.42 mmol) in MeOH (10 ml) at 40°C for 21 h gave the title compound (90 mg, 59%) as a yellow solid after purification by neutral pre-HPLC.

1H NMR (DMSO-66, 500 MHz): δ [ppm] = 12.17 (s, 1H), 8.71 (t, J=4.8 Hz, 1H), 8.16 (s, 1H), 8.14 (s, 1H), 7.51 (s, 1H), 7.42 (s, 2H), 7.12 (d, J=4.9 Hz, 2H), 4.52 ( d, J=4.9 Hz, 2H), 3.97 (s, 2H), 3.17 (d, J=4.9 Hz, 2H), 2.98 (t, J=6.8 Hz, 2H), 2.27 (s, 3H), 2.24 (s, 3H).

HPLC-MS (method B): [m/z]: 421.2 [M+H] ⁺ .

2-{2-[(1H-1,3-Benzodiazol-2-ylmethyl)amino]ethyl}-N-(pyrimidin-2-ylmethyl)-1,3-thiazol-4carboxamide (illustrative example No. 61)

Similar to general procedure 2, tert-butyl 2-({[(tert-butoxy)carbonyl](2-{4-[(pyrimidin-2ylmethyl)carbamoyl]-1,3-thiazol-2-yl}ethyl)amino}methyl )-1H-1,3-benzodiazol-1-carboxylate (38) (86 mg, 0.145 mmol) and 12M HC (0.282 ml, 3.378 mmol) in MeOH (2 ml) at 40°C for 24 h gave the indicated in title compound (24 mg, 42%) as a brown solid after purification by flash column chromatography (gradient elution with 100% DCM, 90% DCM: 10% MeOH and 90% DCM: 10% ammonia in methanol) followed by basic prep-HPLC.

1H NMR (DMSO-J6, 500 MHz): δ [ppm] = 12.18 (s, 1H), 8.75 (dd, J=10.2, 5.4 Hz, 3H), 8, 13 (s, 1H), 7.48 (d, J=36.0 Hz, 2H), 7.39 (t, J=4.9 Hz, 1H), 7.19-7.03 (m, 2H ), 4.66 (d, J=5.8 Hz, 2H), 3.97 (s, 2H), 3.20 (t, J=6.8 Hz, 2H), 2.99 (t, J =6.8 Hz, 2H), 2.66 (s, 1H).

HPLC-MS (method B): [m/z]: 394.2 [M+H] ⁺ .

2- {2-[(1H-1,3-Benzodiazol-2-ylmethyl)amino]ethyl} -N-{[2-(4-methylpiperazin-1-yl)phenyl]methyl} 1,3-thiazol-4 -carboxamide (illustrative example No. 68)

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Similar to general procedure 2, t-butyl N-(1H-1,3-benzodiazol-2-ylmethyl)-N-{2-[4-({[2-(4methylpiperazin-1-yl)phenyl]methyl}carbamoyl) -1,3-thiazol-2-yl]ethyl}carbamate (39) (13.4 mg, 0.02 mmol) in dioxane (0.5 ml) and 4M HC solution in dioxane (55.1 µl) at 50 °C over 16 h gave the title compound (11.5 mg, 85%) as a white solid after trituration in dichloromethane.

¹ H NMR (Methanol-0 ₄ , 500 MHz): δ [ppm] = 8.28 (s, 1H), 7.88 (dt, J=6.6, 3.3 Hz, 2H), 7.69 (dt, J=6.3, 3.3 Hz, 2H), 7.45 (t, J=7.6 Hz, 1H), 7.33 (d, J=6.7 Hz, 2H ), 7.20-7.14 (m, 1H), 5.06 (s, 2H), 4.78 (s, 2H), 3.87 (t, J=6.2 Hz, 2H), 3 .72-3.59 (m, 4H), 3.49 (t, J=10.5 Hz, 2H), 3.37 (s, 2H), 3.27 (d, J=11.0 Hz, 2H), 3.02 (s, 3H).

HPLC-MS (Method D): [m/z]: 490.3 [M+H] ⁺ .

2-{2-[(1H-1,3-Benzodiazol-2-ylmethyl)amino]ethyl}-N-[(2,6-difluorophenyl)methyl]-1,3-thiazole-4carboxamide (illustrative example No. 69)

Similar to general procedure 2, tert-butyl N-(1H-1,3-benzodiazol-2-ylmethyl)-N-[2-(4-{[(2,6difluorophenyl)methyl]carbamoyl}-1,3-thiazol- 2-yl)ethyl]carbamate (40) (90 mg, 0.17 mmol) in dioxane (2 mL) and 4M HC in dioxane (427 μL) at room temperature for 16 h gave the title compound (15 mg , 20.6%) as an off-white solid after purification by preparative HPLC.

¹ H NMR (Methanol-J4, 500 MHz): δ [ppm] = 8.06 (s, 1H), 7.53 (dd, J=5.8, 3.2 Hz, 2H), 7 .34 (tt, J=8.4, 6.5 Hz, 1H), 7.29-7.16 (m, 2H), 7.03-6.88 (m, 2H), 4.68 (s , 2H), 4.07 (s, 2H), 3.23 (t, J=6.6 Hz, 2H), 3.09 (t, J=6.6 Hz, 2H).

HPLC-MS (method A): [m/z]: 428.1 [M+H]+.

2-{2-[(1H-1,3-Benzodiazol-2-ylmethyl)amino]ethyl}-N-{[2-(dimethylamino)phenyl]methyl}-1,3-thiazole4-carboxamide (Illustrative Example No. 70 )

Similar to general procedure 2, tert-butyl N-(1H-1,3-benzodiazol-2-ylmethyl)-N-{2-[4-({[2(dimethylamino)phenyl]methyl}carbamoyl)-1,3- thiazol-2-yl]ethyl}carbamate (41) (73 mg, 0.14 mmol) in dioxane (2 mL) and 4M HC in dioxane (341 μL) at 50°C for 16 h gave the title compound (8.7 mg, 14%) after purification by preparative HPLC.

1H NMR (Methanol-04, 500 MHz): δ [ppm] = 8.08 (s, 1H), 7.53 (dd, J=5.9, 3.2 Hz, 2H), 7, 29-7.14 (m, 5H), 7.02 (td, J=7.4, 1.1 Hz, 1H), 4.68 (s, 2H), 4.08 (s, 2H), 3 .25 (t, J=6.7 Hz, 2H), 3.11 (t, J=6.8 Hz, 2H), 2.67 (s, 6H).

HPLC-MS (Method D): [m/z]: 435.3 [M+H] ⁺ .

2-{2-[(1H-1,3-Benzodiazol-2-ylmethyl)amino]ethyl}-N-[(2-cyanophenyl)methyl]-1,3-thiazole-4carboxamide (illustrative example No. 71)

Similar to general procedure 2, tert-butyl N-(1H-1,3-benzodiazol-2-ylmethyl)-N-[2-(4-{[(2cyanophenyl)methyl]carbamoyl}-1,3-thiazol-2- yl)ethyl]carbamate (42) (54 mg, 0.1 mmol) in dioxane (2 mL) and 4M HC in dioxane (261 μL) at 50°C for 12 h gave the title compound (8 mg, 18%) as a yellow solid after purification by preparative HPLC.

1H NMR (Methanol-J4, 500 MHz): δ [ppm] = 8.11 (s, 1H), 7.74-7.69 (m, 1H), 7.60 (td, J=7 .8, 1.2 Hz, 1H),

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7.56-7.50 (m, 3H), 7.43 (t, J=7.6 Hz, 1H), 7.25-7.19 (m, 2H), 4.78 (s, 2H) , 4.09 (s, 2H), 3.27 (t, J=6.6 Hz, 2H),

3.13 (t, J=6.6 Hz, 2H).

2-{2-[(1H-1,3-Benzodiazol-2-ylmethyl)amino]ethyl}-N-{[2-(trifluoromethoxy)phenyl]methyl}-1,3thiazole-4-carboxamide (Illustrative Example No. 72 )

Similar to general procedure 2, tert-butyl K-(1H-1,3-benzodiazol-2-ylmethyl)-K-{2-[4-({[2(trifluoromethoxy)phenyl]methyl}carbamoyl)-1,3- thiazol-2-yl]ethyl}carbamate (43) (110 mg, 0.191 mmol) and 12M solution of HC (0.317 ml, 4.458 mmol) in MeOH (2 ml) at room temperature for 16 h gave the title compound (41 mg, 45%) as a white solid after purification by basic prep-HPLC.

¹ H NMR (DMSO-d6, 500 MHz): δ [ppm] = 12.19 (s, 1H), 8.88 (t, J=6.2 Hz, 1H), 8.15 (s , 1H), 7.48 (d, J=31.2 Hz, 2H), 7.42-7.30 (m, 4H), 7.13 (d, J=4.9 Hz, 2H), 4 .53 (d, J=6.3 Hz, 2H), 3.97 (s, 2H), 3.19 (t, J=6.8 Hz, 2H), 2.99 (t, J=6, 8 Hz, 2H).

HPLC-MS (method A): [m/z]: 476.2 [M+H]+.

2-{2-[(1H-1,3-Benzodiazol-2-ylmethyl)amino]ethyl}-N-{[2-(difluoromethoxy)phenyl]methyl}-1,3thiazole-4-carboxamide (Illustrative Example No. 74 )

Similar to general procedure 2, crude tert-butyl N-(1H-1,3-benzodiazol-2-ylmethyl)-N-{2-[4-({[2(difluoromethoxy)phenyl]methyl}carbamoyl)-1,3 -thiazol-2-yl]ethyl}carbamate (45) (470 mg) and 12M HC (5 ml) in MeOH (5 ml) at 50° C. for 2 h gave the title compound (56 mg, 35%) as a pale yellow solid after purification by flash column chromatography (KP-NH, eluting with a 0-15% MeOH/DCM gradient), followed by prep-HPLC.

1H NMR (DMSO-06, 500 MHz): δ [ppm] = 12.18 (s, 1H), 8.79 (t, J=6.2 Hz, 1H), 8.13 (s, 1H), 7.47 (s, 2H), 7.35-7.28 (m, 2H), 7.25-7.06 (m, 5H), 4.48 (d, J=6.2 Hz , 2H), 3.96 (s, 2H), 3.22-3.17 (m, 2H), 2.98 (t, J=6.8 Hz, 2H).

HPLC-MS (Method D): [m/z]: 458.2 [M+H] ⁺ .

2-{2-[(1H-1,3-Benzodiazol-2-ylmethyl)amino]ethyl}-N-{[2-(morpholine-4-sulfonyl)phenyl]methyl}1,3-thiazol-4-carboxamide (illustrative example #75)

Similar to general procedure 2, crude tert-butyl N-(1H-1,3-benzodiazol-2-ylmethyl)-N-{2-[4-({[2(morpholine-4-sulfonyl)phenyl]methyl}carbamoyl) -1,3-thiazol-2-yl]ethyl}carbamate (46) (440 mg) and 12M HC (5 ml) in MeOH (5 ml) at 50°C for 2 h gave the title compound (67 mg , 42%) as a pale yellow solid after purification by flash column chromatography (KPNH, eluting with a 0-15% MeOH/DCM gradient), followed by prep-HPLC.

1H NMR (DMSO-06, 500 MHz): δ [ppm] = 12.19 (s, 1H), 8.88 (t, J=6.3 Hz, 1H), 8.17 (s, 1H), 7.84 (d, J=7.9 Hz, 1H), 7.66 (t, J=7.6 Hz, 1H), 7.50 (dd, J=11.8, 7.7 Hz, 4H), 7.17-7.06 (m, 2H), 4.81 (d, J=6.2 Hz, 2H), 3.97 (s, 2H), 3.70-3.61 (m, 4H), 3.20 (t, J=6.8 Hz, 2H), 3.10-3.04 (m, 4H), 2.99 (t, J=6.7 Hz, 2H) .

HPLC-MS (Method D): [m/z]: 541.2 [M+H]+.

2-{2-[(1H-1,3-Benzodiazol-2-ylmethyl)amino]ethyl}-N-[2-(pyridin-2-yl)ethyl]-1,3-thiazole-4carboxamide (Illustrative Example No. 76)

Similarly to general procedure 2, dioxane (2 ml) was added to a solution of tert-butyl N-(1H-1,3-111 039916 benzodiazol-2-ylmethyl)-N-[2-(4-{[2-(pyridine-2 -yl)ethyl]carbamoyl}-1,3-thiazol-2-yl)ethyl]carbamate (47) (52 mg, 0.103 mmol) and 4M HC solution in dioxane (257 μl, 1.03 mmol). The reaction mixture was stirred at room temperature for 16 hours to give the title compound (11 mg, 26%) as a brownish solid after purification by neutral prep-HPLC.

¹ H NMR (Methanol-d4, 500 MHz): δ [ppm] = 8.44 (ddd, J=5.0, 1.7, 0.8 Hz, 1H), 8.03 (s, 1H), 7.72 (td, J=7.7, 1.8 Hz, 1H), 7.55 (dd, J=5.8, 3.2 Hz, 2H), 7.33 (d, J = 7.8 Hz, 1H), 7.29-7.19 (m, 3H), 4.09 (s, 2H), 3.74 (t, J=7.1 Hz, 2H), 3.25 (t, J=6.7 Hz, 2H), 3.13-3.05 (m, 4H).

HPLC-MS (method B): [m/z]: 407.1 [M+H] ⁺ .

2-{2-[(1H-1,3-Benzodiazol-2-ylmethyl)amino]ethyl}-N-(3-fluoropyridin-2-yl)-1,3-thiazole-4carboxamide (Illustrative Example No. 77)

Similar to general procedure 2, dioxane (4 ml) was added to a solution of crude tert-butyl N-(1H-1,3benzodiazol-2-ylmethyl)-N-(2-{4-[(3-fluoropyridin-2-yl)carbamoyl ]-1,3-thiazol-2-yl}ethyl) carbamate (48) (705 mg) and 4M HC solution in dioxane (1.56 ml, 6.24 mmol). The reaction mixture was stirred at room temperature for 16 hours to give the title compound (10 mg, 3.9%) as a brownish solid after purification by neutral prep-HPLC.

¹ H NMR (Methanol-d4, 500 MHz): δ [ppm] = 8.37 (s, 1H), 8.29 (dd, J=4.7, 1.0 Hz, 1H), 7 .75 (ddd, J=9.8, 8.4, 1.4 Hz, 1H), 7.52 (dd, J=6.1, 3.2 Hz, 2H), 7.41 (ddd, J =8.4, 4.7, 3.8 Hz, 1H), 7.24 (dd, J=6.1, 3.2 Hz, 2H), 4.44 (s, 2H), 3.56- 3.46 (m, 4H).

HPLC-MS (Method D): [m/z]: 397.1 [M+H] ⁺ .

2-{2-[(1H-1,3-Benzodiazol-2-ylmethyl)amino]ethyl}-N-[(3-fluoro-6-methylpyridin-2-yl)methyl]-1,3thiazol-4-carboxamide (illustrative example #79)

Similar to general procedure 2, a 4M solution of HC in dioxane (0.36 ml) was added to a solution of tert-butyl N(1H-1,3-benzodiazol-2-ylmethyl)-M-[2-(4-{[(3- fluoro-6-methylpyridin-2-yl)methyl]carbamoyl}-1,3thiazol-2-yl)ethyl]carbamate (50) (76 mg, 0.14 mmol) in dioxane (2 ml), and the resulting mixture was stirred at room temperature for 16 hours to give the title compound (24 mg, 39%) as a colorless oil after purification by basic prep-HPLC.

¹ H NMR (Methanol^, 500 MHz): δ [ppm] = 8.09 (s, 1H), 7.55-7.42 (m, 3H), 7.29-7.14 (m , 3H), 4.71 (d, J=1.7 Hz, 2H), 4.09 (s, 2H), 3.27 (t, J=6.7 Hz, 2H), 3.14 (t , J=6.7 Hz, 2H), 2.45 (s, 3H).

HPLC-MS (Method D): [m/z]: 425.2 [M+H]+.

2-{2-[(1H-1,3-Benzodiazol-2-ylmethyl)amino]ethyl}-N-[1-(pyridin-2-yl)ethyl]-1,3-thiazole-4carboxamide (Illustrative Example No. 80)

Similar to general procedure 2, a 4M solution of HC in dioxane (0.39 ml, 1.56 mmol) was added to a solution of tert-butyl N-(1H-1,3-benzodiazol-2-ylmethyl)-N-[2-(4 -{[1-(pyridin-2-yl)ethyl]carbamoyl}-1,3thiazol-2-yl)ethyl]carbamate (51) (78 mg, 0.15 mmol) in dioxane (2 ml), and the resulting mixture stirred at room temperature for 16 h to give the title compound (15 mg, 23%) as a colorless oil after purification by basic prep-HPLC.

¹ H NMR (Methanol-d4, 500 MHz): δ [ppm] = 8.48 (ddd, J=4.9, 1.7, 0.9 Hz, 1H), 8.08 (s, 1H), 7.79 (td, J=7.8, 1.7 Hz, 1H), 7.54 (dd, J=5.7, 3.2 Hz, 2H), 7.45 (d, J =7.8 Hz, 1H), 7.29 (ddd, J=7.5, 4.9, 1.1 Hz, 1H), 7.267.18 (m, 2H), 5.25 (d, J= 7.0 Hz, 1H), 4.11 (s, 2H), 3.29 (t, J=6.7 Hz, 2H), 3.16-3.12 (m, 2H), 1.57 ( e, J=7.0 Hz, 3H).

HPLC-MS (Method D): [m/z]: 407.2 [M+H] ⁺ .

2-{2-[(1H-1,3-Benzodiazol-2-ylmethyl)amino]ethyl}-N-{[6-(trifluoromethyl)pyridin-3-yl]methyl}-1,3thiazol-4-carboxamide ( illustrative example No. 81)

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Similar to general procedure 2, a 4M solution of HC in dioxane (0.41 ml, 1.64 mmol) was added to a solution of tert-butyl N-(1H-1,3-benzodiazol-2-ylmethyl)-N-{2-[4 -({[6-(trifluoromethyl)pyridine-3-

yl]methyl}carbamoyl)-1,3-thiazol-2-yl]ethyl}carbamate (52) (92 mg, 0.16 mmol) in dioxane (2 ml) and the resulting mixture was stirred at room temperature for 16 h to give the title compound (21 mg, 27%) as a colorless oil after purification by basic prep-HPLC.

1H NMR (Methanol-d4, 500 MHz): δ [ppm] = 8.71 (d, J=1.5 Hz, 1H), 8.11 (s, 1H), 8.00 (dd, J=8.1, 1.5 Hz, 1H), 7.77 (d, J=8.1 Hz, 1H), 7.53 (dd, J=5.8, 3.2 Hz, 2H), 7.28-7.14 (m, 2H), 4.68 (s, 2H), 4.09 (s, 2H), 3.27 (t, J=6.7 Hz, 2H), 3.12 (t, J=6.7 Hz, 2H).

HPLC-MS (Method D): [m/z]: 461.1 [M+H]+.

2-{2-[(1H-1,3-Benzodiazol-2-ylmethyl)amino]ethyl}-N-[(3-chloropyridin-2-yl)methyl]-1,3-thiazole-4carboxamide (illustrative example no. 82)

Similar to general procedure 2, tert-butyl N-(1H-1,3-benzodiazol-2-ylmethyl)-N-[2-(4-{[(3chloropyridin-2-yl)methyl]carbamoyl}-1,3- thiazol-2-yl)ethyl]carbamate (53) (161 mg, 0.363 mmol) and 12M HC (1.6 ml, 19.2 mmol) in MeOH (1.6 ml) at 50°C for 4 hours gave the title compound (72 mg, 54%) as a white solid after purification by basic prep-HPLC.

1H NMR (DMSO-d ₆ , 500 MHz): δ [ppm] = 12.22 (s, 1H), 8.71 (t, J=5.5 Hz, 1H), 8.47 (dd , J=4.7, 1.3 Hz, 1H), 8.15 (s, 1H), 7.95 (dd, J=8.1, 1.3 Hz, 1H), 7.51 (s, 1H), 7.45 (s, 1H), 7.37 (dd, J=8.1, 4.7 Hz, 1H), 7.12 (d, J=4.8 Hz, 2H), 4, 67 (d, J=5.5 Hz, 2H), 3.97 (s, 2H), 3.20 (t, J=6.8 Hz, 2H), 2.98 (t, J=6.8 Hz, 2H).

HPLC-MS (Method D): [m/z]: 427.1 [M+H] ⁺ .

2-{2-[(1H-1,3-Benzodiazol-2-ylmethyl)amino]ethyl}-N-[(2-oxo-1,2-dihydropyridin-3-yl)methyl]-1,3thiazol-4 -carboxamide (illustrative example No. 83)

Similar to general procedure 2, crude tert-butyl N-(1H-1,3-benzodiazol-2-ylmethyl)-N-{2-[4-({[2(tert-butoxy)pyridin-3-yl]methyl} carbamoyl)-1,3-thiazol-2-yl]ethyl}carbamate (54) (205 mg, 0.36 mmol) and 12M HC (2 ml) in MeOH (2 ml) at 50°C for 4 h gave the title compound (66 mg, 44%) as a pale yellow solid after purification by basic prep-HPLC.

1H NMR (DMSO-d ₆ , 500 MHz): δ [ppm] = 12.17 (br s, 1H), 11.65 (br s, 1H), 8.59 (t, J= 6.0 Hz, 1H), 8.11 (s, 1H), 7.48 (s, 2H), 7.32-7.26 (m, 1H), 7.24 (d, J=6.7 Hz, 1H), 7.12 (dd, J=6.0, 3.1 Hz, 2H), 6.15 (t, J=6.6 Hz, 1H), 4.22 (d, J=6 .0 Hz, 2H), 3.96 (s, 2H), 3.21-3.14 (m, 2H), 2.97 (t, J=6.8 Hz, 2H).

HPLC-MS (Method D): [m/z]: 409.2 [M+H] ⁺ .

2-{2-[(1H-1,3-Benzodiazol-2-ylmethyl)amino]ethyl}-N-[(1-methyl-1H-imidazol-5-yl)methyl]-1,3thiazol-4-carboxamide (illustrative example #84)

Similar to general procedure 2, tert-butyl N-(1H-1,3-benzodiazol-2-ylmethyl)-N-[2-(4-{[(1-methyl1H-imidazol-5-yl)methyl]carbamoyl}- 1,3-thiazol-2-yl)ethyl]carbamate (55) (148 mg, 0.3 mmol) and 4M HC in dioxane (7 ml) at room temperature for 2 h gave the title compound (58 mg , 49%) as a colorless glass after purification by basic pre-HPLC.

1H NMR (DMSO-d ₆ , 500 MHz): δ [ppm] = 8.66 (t, J=6.0 Hz, 1H), 8.13 (s, 1H), 7.60-7 .31 (m, 3H), 7.286.97 (m, 2H), 6.79 (s, 1H), 4.41 (d, J=6.0 Hz, 2H), 3.96 (s, 2H) , 3.60 (s, 3H), 3.16 (t, J=6.8 Hz, 2H), 2.97 (t, J=6.8 Hz, 2H).

HPLC-MS (method B): [m/z]: 396.2 [M+H] ⁺ .

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2-{2-[(1H-1,3-Benzodiazol-2-ylmethyl)amino]ethyl}-N-(1,3-oxazol-2-ylmethyl)-1,3-thiazole-4carboxamide (Illustrative Example No. 85 )

Similar to general procedure 2, a 4M solution of HC in dioxane (0.49 mL, 1.96 mmol) was added to a solution of tert-butyl N-(1H-1,3-benzodiazol-2-ylmethyl)-N-(2-{4 -[(1,3-oxazol-2-ylmethyl)carbamoyl]-1,3thiazol-2-yl}ethyl)carbamate (56) (95 mg, 0.196 mmol) in dioxane (2 ml) and the resulting mixture was stirred at room temperature. temperature for 3 hours to give the title compound (13 mg, 17%) as a pale yellow oil after purification by neutral prep-HPLC.

1H NMR (Methanol-J4, 500 MHz): δ [ppm] = 8.11 (s, 1H), 7.87 (d, J=0.8 Hz, 1H), 7.54 (dd, J=5.9, 3.2 Hz, 2H), 7.27-7.18 (m, 2H), 7.13 (d, J=0.8 Hz, 1H), 4.70 (s, 2H ), 4.10 (s, 2H), 3.27 (t, J=6.6 Hz, 2H), 3.13 (t, J=6.6 Hz, 2H).

HPLC-MS (Method D): [m/z]: 383.1 [M+H] ⁺ .

2-{2-[(1H-1,3-Benzodiazol-2-ylmethyl)amino]ethyl}-N-[(1-methyl-1H-pyrazol-3-yl)methyl]-1,3-thiazol4- carboxamide (illustrative example No. 86)

Similar to general procedure 2, a 4M solution of HC in dioxane (0.27 ml, 1.08 mmol) was added to raster-butyl solution

N-(1H-1,3-benzodiazol-2-ylmethyl)-N -[2-(4-{[(1-methyl-1 H-pyrazol-3 yl)methyl]carbamoyl}-1,3-thiazol- 2-yl)ethyl]carbamate (57) (53 mg, 0.11 mmol) in dioxane (2 ml) at room temperature for 3 h to give the title compound (16 mg, 38%) as a clear oil after purification by neutral prep-HPLC.

¹ H NMR (Methanol-D4, 500 MHz): δ [ppm] = 8.08 (s, 1H), 7.57-7.46 (m, 3H), 7.28-7.15 ( m, 2H), 6.22 (d, J=2.2 Hz, 1H), 4.55 (s, 2H), 4.08 (s, 2H), 3.84 (s, 3H), 3, 25 (t, J=6.7 Hz, 2H), 3.10 (t, J=6.7 Hz, 2H).

HPLC-MS (Method D): [m/z]: 396.2 [M+H]+.

2-{2-[(1H-1,3-Benzodiazol-2-ylmethyl)amino]ethyl}-N-(pyridazin-3-ylmethyl)-1,3-thiazol-4carboxamide (illustrative example No. 87)

Similar to general procedure 2, a 4M solution of HC in dioxane (0.51 ml, 2.04 mmol) was added to a solution of tert-butyl N-(1H-1,3-benzodiazol-2-ylmethyl)-N-(2-{4 -[(pyridazin-3-ylmethyl)carbamoyl]-1,3thiazol-2-yl}ethyl)carbamate (58) (101 mg, 0.2 mmol) in dioxane (2 ml) at 50° C. for 1 hour to give the title compound (15 mg, 19%) as a brownish solid after purification by neutral prep-HPLC.

1H NMR (Methanol-04, 500 MHz): δ [ppm] = 9.10 (dd, J=4.8, 1.7 Hz, 1H), 8.12 (s, 1H), 7, 76-7.64 (m, 2H), 7.53 (dd, J=6.0, 3.2 Hz, 2H), 7.22 (dd, J=6.0, 3.2 Hz, 2H) , 4.89 (s, 2H), 4.10 (s, 2H), 3.28 (t, J=6.6 Hz, 2H), 3.13 (t, J=6.6 Hz, 2H) .

HPLC-MS (Method D): [m/z]: 394.1 [M+H] ⁺ .

2-{2-[(1H-1,3-Benzodiazol-2-ylmethyl)amino]ethyl}-N-[(1-methyl-1H-pyrazol-5-yl)methyl]-1,3-thiazol4- carboxamide (illustrative example No. 88)

Similar to general procedure 2, a 4M solution of HC in dioxane (0.26 mL, 1.04 mmol) was added to a solution of tert-butyl N-(1H-1,3-benzodiazol-2-ylmethyl)-N-[2-(4 -{[(1-methyl-1H-pyrazol-5yl)methyl]carbamoyl}-1,3-thiazol-2-yl)ethyl]carbamate (59) (51 mg, 0.103 mmol) in dioxane (2 ml) at 50° C. for 1 hour to give the title compound (12 mg, 30%) as an orange oil after purification by neutral prep-HPLC.

1H NMR (Methanol-J4, 500 MHz): δ [ppm] = 8.10 (s, 1H), 7.53 (dd, J=6.0, 3.2 Hz, 2H), 7, 36 (d, J=1.9 Hz, 1H), 7.23 (dd, J=6.0, 3.2 Hz, 2H), 6.25 (d, J=1.9 Hz, 1H), 4.63 (s, 2H), 4.08 (s, 2H), 3.89 (s, 3H), 3.26 (t, J=6.7

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Hz, 2H), 3.11 (t, J=6.7 Hz, 2H).

HPLC-MS (Method D): [m/z]: 396.2 [M+H]+.

2-{2-[(1H-1,3-Benzodiazol-2-ylmethyl)amino]ethyl}-N-[(3-fluoropyridin-4-yl)methyl]-1,3-thiazole-4carboxamide (illustrative example no. 89)

Similar to general procedure 2, tert-butyl N-(1H-1,3-benzodiazol-2-ylmethyl)-N-[2-(4-{[(3fluoropyridin-4-yl)methyl]carbamoyl}-1,3- thiazol-2-yl)ethyl]carbamate (60) (137 mg, 0.191 mmol, 71% pure) and 12M HC (1.4 ml, 16.8 mmol) in MeOH (1.4 ml) at 50°C in gave the title compound (46 mg, 59%) as a pale yellow solid over 2 hours after purification by neutral prep-HPLC.

¹ H NMR (DMSO-d6, 500 MHz): δ [ppm] = 12.18 (s, 1H), 8.99 (t, J=6.1 Hz, 1H), 8.49 (d , J=1.6 Hz, 1H), 8.35 (d, J=4.8 Hz, 1H), 8.15 (s, 1H), 7.48 (br s, 2H), 7.35 -7.26 (m, 1H), 7.12 (d, J=3.5 Hz, 2H), 4.52 (d, J=6.1 Hz, 2H), 3.97 (s, 2H) , 3.19 (t, J=6.8 Hz, 2H), 2.98 (t, J=6.8 Hz, 2H).

HPLC-MS (method B): [m/z]: 411.1 [M+H] ⁺ .

2-{2-[(1H-1,3-Benzodiazol-2-ylmethyl)amino]ethyl}-N-[(3-methylpyridin-4-yl)methyl]-1,3-thiazole-4carboxamide trihydrochloride (illustrative example) No. 90)

Similar to general procedure 2, a 4M solution of HC in dioxane (1.8 ml) was added to a solution of tert-butyl N-(1H-1,3-benzodiazol-2-ylmethyl)-N-[2-(4-{[(3 -methylpyridin-4-yl)methyl]carbamoyl}-1,3-thiazol-2yl)ethyl]carbamate (61) (363 mg, 0.72 mmol) in dioxane (5 mL) at room temperature for 16 hours to give the title compound (203 mg, 55%) as a white solid. This solid was obtained by precipitation from a mixture of DCM/MeOH with the addition of heptane, followed by washing with diethyl ether.

¹ H NMR (Methanol-d4, 500 MHz): δ [ppm] = 8.69 (s, 1H), 8.60 (d, J=6.1 Hz, 1H), 8.30 (s , 1H), 8.00 (d, J=6.1 Hz, 1H), 7.83 (dt, J=6.5, 3.3 Hz, 2H), 7.61 (dt, J=6, 5, 3.3 Hz, 2H), 5.00 (s, 2H), 3.89 (t, J=6.3 Hz, 2H), 3.69 (t, J=6.3 Hz, 2H) , 2.63 (s, 3H) (CH ₂ signal overlapping with solvent signal).

HPLC-MS (method B): [m/z]: 407.2 [M+H] ⁺ .

2-{2-[(1H-1,3-Benzodiazol-2-ylmethyl)amino]ethyl}-N-[(1-methyl-1H-pyrazol-4-yl)methyl]-1,3-thiazol4- carboxamide (illustrative example No. 91) o

Similar to general procedure 2, tert-butyl N-(1H-1,3-benzodiazol-2-ylmethyl)-N-[2-(4-{[(1-methyl1H-pyrazol-4-yl)methyl]carbamoyl}- 1,3-thiazol-2-yl)ethyl]carbamate (62) (125 mg, 0.25 mmol) and 12M HC (0.49 ml, 5.88 mmol) in MeOH (5 ml) at 50°C in gave the title compound (42 mg, 42%) as a tan solid over 2 hours after purification by basic prep-HPLC.

1H NMR (DMSO-d6, 500 MHz): δ [ppm] = 12.16 (s, 1H), 8.55 (t, J=6.0 Hz, 1H), 8.08 (s, 1H), 7.55 (s, 1H), 7.48 (d, J=38.4 Hz, 2H), 7.32 (s, 1H), 7.20-7.03 (m, 2H), 4.25 (d, J=6.0 Hz, 2H), 3.95 (s, 2H), 3.76 (s, 3H), 3.15 (t, J=6.8 Hz, 2H), 2.95 (t, J=6.8 Hz, 2H).

HPLC-MS (Method D): [m/z]: 396.2 [M+H] ⁺ .

2-{2-[(1H-1,3-Benzodiazol-2-ylmethyl)amino]ethyl}-N-[(6-methylpyridazin-3-yl)methyl]-1,3-thiazole-4carboxamide (Illustrative Example No. 93) about

Similar to general procedure 2, crude tert-butyl N-(1H-1,3-benzodiazol-2-ylmethyl)-N-[2-(4-{[(6methylpyridazin-3-yl)methyl]carbamoyl}-1,3 -thiazol-2-yl)ethyl]carbamate (63) (99mg, 0.154mmol, 79% pure) and 12M HC (1mL) in MeOH (1mL) at 50°C for 2h gave the title compound (32 mg, 51%) as a light brown solid after purification by neutral prep 115 039916

HPLC.

1H NMR (DMSO-0b, 500 MHz): δ [ppm] = 12.18 (s, 1H), 9.01 (t, J=6.1 Hz, 1H), 8.14 (s, 1H), 7.57-7.39 (m, 4H), 7.13 (s, 2H), 4.70 (d, J=6.1 Hz, 2H), 3.96 (s, 2H), 3.19 (t, J=6.8 Hz, 2H), 2.98 (t, J=6.8 Hz, 2H), 2.58 (s, 3H).

HPLC-MS (method B): [m/z]: 408.2 [M+H]+.

2-{2-[(1H-1,3-Benzodiazol-2-ylmethyl)amino]ethyl}-N-(1H-imidazol-2-ylmethyl)-1,3-thiazole-4carboxamide (illustrative example No. 95) o < Ύ ^Ν Ίι w__

V-NH ^H U' \ ⁿ m_^[IJ

Η

Similar to general procedure 2, a 4M solution of HC in dioxane (1 mL, 4 mmol) was added to a solution of tert-butyl N-(1H-1,3-benzodiazol-2-ylmethyl)-N-(2-{4-[(1H -imidazol-2-ylmethyl)carbamoyl]-1,3-thiazol2-yl}ethyl)carbamate (64) (61 mg, 0.13 mmol) in dioxane (4 mL) at room temperature for 18 h to give title compound (23 mg, 48%) as a yellow solid after purification by basic prep-HPLC.

1H NMR (DMSO-06, 500 MHz): δ [ppm] = 12.19 (s, 1H), 11.77 (s, 1H), 8.61 (t, J=5.9 Hz, 1H), 8.14 (s, 1H), 7.49 (br s, 2H), 7.13 (dd, J=5.9, 2.8 Hz, 2H), 7.00 (br s , 1H), 6.81 (brs, 1H), 4.48 (d, J=5.9 Hz, 2H), 3.97 (s, 2H), 3.17 (d, J=6, 2 Hz, 2H), 2.98 (t, J=6.8 Hz, 2H).

HPLC-MS (method B): [m/z]: 382.1 [M+H] ⁺ .

2-{2-[(1H-1,3-Benzodiazol-2-ylmethyl)amino]ethyl}-N-{[2-(morpholin-4-yl)pyridin-4-yl]methyl}1,3-thiazole -4-carboxamide (illustrative example No. 96)

Similar to general procedure 2, a 4M solution of HC in dioxane (1 ml) was added to a solution of tert-butyl N(1H-1,3-benzodiazol-2-ylmethyl)-N-{2-[4-({[2-(morpholine -4-yl)pyridin-4-yl]methyl}carbamoyl)-1,3thiazol-2-yl]ethyl}carbamate (65) (108 mg, 0.19 mmol) in dioxane (4 ml) at room temperature for 15 hours to give the title compound (46 mg, 51%) as an orange solid after purification by neutral prep-HPLC.

1H NMR (DMSO-06, 500 MHz): δ [ppm] = 8.90 (t, J=6.3 Hz, 1H), 8.15 (s, 1H), 8.03 (d, J=5.1 Hz, 1H), 7.50 (dd, J=5.7, 3.3 Hz, 2H), 7.14 (d.s., J=7.1, 3.9 Hz, 2H), 6.73 (s, 1H), 6.61 (d, J=5.1 Hz, 1H), 4.38 (d, J=6.3 Hz, 2H), 4.15 (d, J=29.7 Hz, 1H), 4.05 (s, 2H), 3.72-3.62 (m, 4H), 3.23 (t, J=6.8 Hz, 2H), 3, 18 (s, 2H) 3.07 (t, J=6.8 Hz, 2H).

HPLC-MS (method B): [m/z]: 478.2 [M+H] ⁺ .

2-{2-[(III-1,3-Benzodiazol-2-ylmethyl)amino]ethyl}-N-[(5-methylpyridin-2-yl)methyl]-1,3-thiazole-4carboxamide (illustrative example no. 97)

Similar to general procedure 2, a 4M solution of HC in dioxane (0.24 ml, 0.96 mmol) was added to a solution of tert-butyl N-(1H-1,3-benzodiazol-2-ylmethyl)-N-[2-(4 -{[(5-methylpyridin-2-yl)methyl]carbamoyl}1,3-thiazol-2-yl)ethyl]carbamate (66) (48 mg, 0.095 mmol) in dioxane (2 ml) at room temperature for 12 hours to give the title compound (10 mg, 26%) as a colorless oil after purification by neutral prep-HPLC.

1H NMR (Methanol-b ₄ , 500 MHz): δ [ppm] = 8.31 (s, 1H), 8.10 (s, 1H), 7.60 (dd, J=8.0, 1.8 Hz, 1H), 7.52 (dd, J=6.0, 3.1 Hz, 2H), 7.29 (d, J=8.0 Hz, 1H), 7.22 (dd, J=6.0, 3.1 Hz, 2H), 4.65 (s, 2H), 4.10 (s, 2H), 3.28 (t, J=6.6 Hz, 2H), 3, 13 (t, J=6.6 Hz, 2H), 2.33 (s, 3H).

HPLC-MS (Method D): [m/z]: 407.1 [M+H] ⁺ .

2-{2-[(1H-1,3-Benzodiazol-2-ylmethyl)amino]ethyl}-N-{[6-(dimethylamino)pyridin-3-yl]methyl}-1,3thiazol-4-carboxamide ( illustrative example No. 98)

- 116 039916

Similar to general procedure 2, a 4M solution of HC in dioxane (0.17 ml, 0.68 mmol) was added to a solution of tert-butyl N-(1H-1,3-benzodiazol-2-ylmethyl)-N-{2-[4 -({[6-(dimethylamino)pyridin-3yl]methyl}carbamoyl)-1,3-thiazol-2-yl]ethyl}carbamate (67) (36 mg, 0.07 mmol) in dioxane (2 ml) at room temperature for 12 hours to give the title compound (8 mg, 27%) as a colorless oil after purification by neutral prep-HPLC.

1H NMR (Methanol-d4, 500 MHz): δ [ppm] = 8.07 (s, 1H), 8.05 (d, J=2.1 Hz, 1H), 7.57-7, 50 (m, 3H), 7.267.20 (m, 2H), 6.61 (d, J=8.8 Hz, 1H), 4.43 (s, 2H), 4.07 (s, 2H), 3.24 (t, J=6.6 Hz, 2H), 3.10 (t, J=6.6 Hz, 2H), 3.06 (s, 6H).

HPLC-MS (method B): [m/z]: 436.3 [M+H]+.

2-{2-[(1H-1,3-Benzodiazol-2-ylmethyl)amino]ethyl}-N-[(2-methylpyridin-4-yl)methyl]-1,3-thiazole-4carboxamide (Illustrative Example No. 99) about

Similar to general procedure 2, a 4M solution of HC in dioxane (0.18 mL, 0.72 mmol) was added to a solution of tert-butyl N-(1H-1,3-benzodiazol-2-ylmethyl)-N-[2-(4 -{[(2-methylpyridin-4-yl)methyl]carbamoyl}1,3-thiazol-2-yl)ethyl]carbamate (68) (36 mg, 0.07 mmol) in dioxane (2 ml) at room temperature over 12 hours to give the title compound (20 mg, 69%) as a colorless oil after purification by neutral prep-HPLC.

¹ H NMR (Methanol-d4, 500 MHz): δ [ppm] = 8.31 (d, J=5.3 Hz, 1H), 8.11 (s, 1H), 7.52 (dd , J=5.9, 3.2 Hz, 2H), 7.30-7.13 (m, 4H), 4.58 (s, 2H), 4.09 (s, 2H), 3.28 ( t, J=6.6 Hz, 2H), 3.13 (t, J=6.6 Hz, 2H), 2.49 (s, 3H).

HPLC-MS (method B): [m/z]: 407.2 [M+H] ⁺ .

2-{2-[(1H-1,3-Benzodiazol-2-ylmethyl)amino]ethyl}-N-[(1,5-dimethyl-1H-pyrazol-4-yl)methyl]-1,3thiazol-4 -carboxamide (illustrative example No. 100)

Similar to general procedure 2, a 4M solution of HC in dioxane (0.36 mL, 1.44 mmol) was added to a solution of tert-butyl N-(1H-1,3-benzodiazol-2-ylmethyl)-N-[2-(4 -{[(1,5-dimethyl-1H-pyrazol-4yl)methyl]carbamoyl}-1,3-thiazol-2-yl)ethyl]carbamate (69) (74 mg, 0.15 mmol) in dioxane (2 ml) at room temperature for 12 hours to give the title compound (19 mg, 33%) as a colorless oil after purification by neutral prep-HPLC.

¹ H NMR (Methanol-d4, 500 MHz): δ [ppm] = 8.05 (s, 1H), 7.53 (dd, J=5.8, 3.2 Hz, 2H), 7 .38 (s, 1H), 7.307.17 (m, 2H), 4.38 (s, 2H), 4.07 (s, 2H), 3.74 (s, 3H), 3.23 (t, J=6.7 Hz, 2H), 3.09 (t, J=6.7 Hz, 2H), 2.29 (s, 3H).

HPLC-MS (Method D): [m/z]: 410.2 [M+H] ⁺ .

2- {2-[(1H-1,3-Benzodiazol-2-ylmethyl)amino]ethyl}-N-{[3-chloro-5-(trifluoromethyl)pyridin-2yl]methyl}-1,3-thiazol- 4-carboxamide (illustrative example No. 101)

Similar to general procedure 2, tert-butyl N-(1H-1,3-benzodiazol-2-ylmethyl)-N-{2-[4-({[3-chloro-5(trifluoromethyl)pyridin-2-yl]methyl }carbamoyl)-1,3-thiazol-2-yl]ethyl}carbamate (70) (216 mg, 0.163 mmol, 45% pure) and 12M HC (2.1 ml) in MeOH (2.1 ml) at 50 °C over 2 h gave the title compound (67 mg, 80%) as a white solid after purification by basic pre-HPLC.

¹ H NMR (DMSO-dg, 500 MHz): δ [ppm] = 12.18 (s, 1H), 8.91-8.86 (m, 1H), 8.77 (t, J= 5.6 Hz, 1H), 8.45 (d, J=1.5 Hz, 1H), 8.14 (s, 1H), 7.59-7.35 (m, 2H), 7.19- 7.05 (m, 2H), 4.75 (d, J=5.6 Hz, 2H), 3.97 (s, 2H), 3.20 (t, J=6.8 Hz, 2H), 2.99 (t, J=6.8 Hz, 2H).

HPLC-MS (Method D): [m/z]: 495.0 [M+H] ⁺ .

2-{2-[(1H-1,3-Benzodiazol-2-ylmethyl)amino]ethyl}-X-[(3-chloro-5-fluoropyridin-2-yl)methyl]-1,3thiazol-4-carboxamide (illustrative example #102)

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Similar to general procedure 2, tert-butyl X-(1H-1,3-benzodiazol-2-ylmethyl)-X-[2-(4-{[(3-chloro-5fluoropyridin-2-yl)methyl]carbamoyl}- 1,3-thiazol-2-yl)ethyl]carbamate (71) (157 mg, 76%, 62% pure) and 12 M HC (1.6 ml) in MeOH (1.6 ml) at 50°C in gave the title compound (50 mg, 62%) as a white solid over 2 hours after purification by basic prep-HPLC.

1H NMR (DMSO-d ₆ , 500 MHz): δ [ppm] = 12.17 (s, 1H), 8.67 (t, J=5.6 Hz, 1H), 8.54 (d , J=2.5 Hz, 1H), 8.14 (s, 1H), 8.11 (dd, J=8.5, 2.5 Hz, 1H), 7.60-7.36 (m, 2H), 7.12 (d, J=4.7 Hz, 2H), 4.64 (d, J=5.1 Hz, 2H), 3.97 (s, 2H), 3.19 (t, J=6.8 Hz, 2H), 2.98 (t, J=6.8 Hz, 2H).

HPLC-MS (Method D): [m/z]: 445.1 [M+H] ⁺ .

2-{2-[(1H-1,3-Benzodiazol-2-ylmethyl)amino]ethyl}-N-[(2-fluoropyridin-3-yl)methyl]-1,3-thiazole-4carboxamide (illustrative example no. 103)

Similar to general procedure 2, tert-butyl X-(1H-1,3-benzodiazol-2-ylmethyl)-X-[2-(4-{[(2fluoropyridin-3-yl)methyl]carbamoyl}-1,3- thiazol-2-yl)ethyl]carbamate (72) (127 mg, 0.249 mmol) and a 4M solution of HC in dioxane (0.622 ml, 2.487 mmol) in dioxane (4.4 ml) at room temperature for 5 hours gave the indicated in title compound (26 mg, 25%) as a yellow solid after purification by basic prep-HPLC.

1H NMR (DMSO-d ₆ , 500 MHz): δ [ppm] = 8.94 (t, J=6.2 Hz, 1H), 8.17 (s, 1H), 8.13-8 .09 (m, 1H), 7.857.79 (m, 1H), 7.55-7.46 (m, 2H), 7.32-7.28 (m, 1H), 7.18-7.12 (m, 2H), 4.47 (d, J=6.1 Hz, 2H), 4.11-4.08 (m, 2H), 3.27-3.23 (m, 2H), 3, 15-3.08 (wide m, 2H).

HPLC-MS (method F): [m/z]: 411.2 [M+H]+.

2-{2-[(1H-1,3-Benzodiazol-2-ylmethyl)amino]ethyl}-N-[(2-methoxypyridin-4-yl)methyl]-1,3-thiazole-4carboxamide (illustrative example no. 104)

Similar to general procedure 2, tert-butyl X-(1H-1,3-benzodiazol-2-ylmethyl)-N-[2-(4-{[(2methoxypyridin-4-yl)methyl]carbamoyl}-1,3- thiazol-2-yl)ethyl]carbamate (73) (104mg, 0.199mmol, 96% pure) and 12M HC (1mL) in MeOH (1mL) at 50°C for 2h gave the title compound ( 35 mg, 42%) as a pale yellow solid after purification by basic prep-HPLC.

¹ H NMR (DMSO-66, 500 MHz): δ [ppm] = 12.17 (s, 1H), 8.95 (t, J=6.3 Hz, 1H), 8.13 (s , 1H), 8.06 (d, J=5.3 Hz, 1H), 7.58-7.38 (m, 2H), 7.17-7.07 (m, 2H), 6.89 ( dd, J=5.3, 1.1 Hz, 1H), 6.65 (s, 1H), 4.40 (d, J=6.3 Hz, 2H), 3.96 (s, 2H), 3.81 (s, 3H), 3.19 (t, J=6.8 Hz, 2H), 2.98 (t, J=6.8 Hz, 2H).

HPLC-MS (method B): [m/z]: 423.1 [M+H] ⁺ .

2-{2-[(1H-1,3-Benzodiazol-2-ylmethyl)amino]ethyl}-X-[(4,6-dimethylpyridin-3-yl)methyl]-1,3thiazole-4-carboxamide (illustrative example no. 105)

Similar to general procedure 2, tert-butyl X-(1H-1,3-benzodiazol-2-ylmethyl)-X-[2-(4-{[(4,6dimethylpyridin-3-yl)methyl]carbamoyl}-1, 3-thiazol-2-yl)ethyl]carbamate (74) (0.077 g, 0.148 mmol) and 12M HC (0.287 ml, 3.449 mmol) in MeOH (5 ml) at 45°C for 20 h gave the title compound (25 mg, 40%) as a yellow solid after purification by neutral pre-HPLC.

1H NMR (DMSO-a ₆ , 500 MHz): δ [ppm] = 8.73 (t, J=6.1 Hz, 1H), 8.25 (s, 1H), 8.11 (s , 1H), 7.56-7.37 (br.m, 2H), 7.16-7.09 (m, 2H), 7.02 (s, 1H), 4.42 (d, J=6 .0 Hz, 2H), 3.95 (s, 2H), 3.16 (t, J=6.5 Hz, 5H), 2.96 (t, J=6.8 Hz, 2H), 2, 37 (s, 3H), 2.28 (s, 3H).

HPLC-MS (method d): [m/z]: 421.2 [M+H] ⁺ .

2-{2-[(1H-1,3-Benzodiazol-2-ylmethyl)amino]ethyl}-N-[(4-methylpyridin-2-yl)methyl]-1,3-thiazol-4- 118 039916 carboxamide (illustrative example #106)

Similar to general procedure 2, tert-butyl N(1H-1,3-benzodiazol-2-ylmethyl)-^[2-(4-{[(4methylpyridin-2-yl)methyl]carbamoyl}-1,3-thiazol- 2-yl)ethyl]carbamate (75) (90 mg, 0.178 mmol) and 12M HC (0.345 ml, 4.143 mmol) in MeOH (5 ml) at 45°C for 20 h gave the title compound (21 mg, 29%) as a yellow solid after purification by neutral prep-HPLC.

1H NMR (DMSO-J6, 500 MHz): δ [ppm] = 8.82 (t, J=6.0 Hz, 1H), 8.34 (d, J=5.0 Hz, 1H) , 8.14 (s, 1H), 7.57-7.39 (m, 2H), 7.14-7.10 (m, 3H), 7.09 (d, J=5.1 Hz, 1H ), 4.52 (d, J=6.0 Hz, 2H), 3.97 (s, 2H), 3.19 (t, J=6.8 Hz, 3H), 3.01-2.96 (m, 2H), 2.27 (s, 3H).

HPLC-MS (method d): [m/z]: 407.2 [M+H] ⁺ .

2-{2-[(1H-1,3-Benzodiazol-2-ylmethyl)amino]ethyl}-X-[(3,5-difluoropyridin-2-yl)methyl]-1,3-thiazole4-carboxamide (illustrative example no. 108)

Similar to general procedure 2, tert-butyl N(1H-1,3-benzodiazol-2-ylmethyl)-N[2-(4-{[(3,5difluoropyridin-2-yl)methyl]carbamoyl}-1,3- thiazol-2-yl)ethyl]carbamate (77) (112 mg, 0.21 mmol) and a 4M solution of HC in dioxane (0.53 ml, 2.119 mmol) in dioxane (2 ml) at room temperature for 16 h gave the title compound (36.7 mg, 40%) as a colorless oil after purification by neutral prep-HPLC.

1H NMR (Methanol ₄ , 500 MHz): δ [ppm] = 8.27 (d, J=2.3 Hz, 1H), 8.09 (s, 1H), 7.61-7 .55 (m, 1H), 7.547.49 (m, 2H), 7.25-7.16 (m, 2H), 4.74 (s, 2H), 4.10 (s, 2H), 3. 28 (t, J=6.6 Hz, 2H), 3.14 (t, J=6.6 Hz, 2H).

HPLC-MS (method B): [m/z]: 429.2 [M+H]+.

2-{2-[(1H-1,3-Benzodiazol-2-ylmethyl)amino]ethyl}-N{[3-(hydroxymethyl)pyridin-2-yl]methyl}1,3-thiazol-4-carboxamide ( illustrative example No. 110)

Similar to general procedure 2, a 4M solution of HC in dioxane (3.28 ml, 13.13 mmol) was added to a solution of tert-butyl N-(1H-1,3-benzodiazol-2-ylmethyl)-N-[2-(4 - {[(3-{[(tert-butyldimethylsilyl)oxy]methyl}pyridin-2-yl)methyl]carbamoyl}-1,3-thiazol-2-yl)ethyl]carbamate (78) (191.5 mg, 0.09 mmol, 30% pure) in dioxane (2 mL) at room temperature for 15 h to give the title compound (8.9 mg, 23.5%) as a yellow oil after purification by basic prep-HPLC. .

1H NMR (DMSO-J6, 500 MHz): δ [ppm] = 12.21 (s, 1H), 8.75 (t, J=5.0 Hz, 1H), 8.40 (dd, J=4.8, 1.4 Hz, 1H), 8.14 (s, 1h), 7.81-7.77 (m, 1H), 7.57-7.40 (breadth m, 2H) , 7.31 (dd, J=7.6, 4.8 Hz, 1H), 7.14-7.10 (m, 2H), 5.41 (brs, 1H), 4.63-4 .59 (m, 4H), 3.97 (s, 2H), 3.20 (t, J=6.8 Hz, 2H), 2.98 (t, J=6.8 Hz, 2H).

HPLC-MS (method B): [m/z]: 422.2 [M+H] ⁺ .

The above general scheme 2:

2-{2-[(1Н-1,3-benzodiazol-2-ylmethyl)amino]ethyl}-1,3-thiazole-4-carboxylic acid (80)

Similar to general procedure 5, LiOH (0.142 g, 5.92 mmol) and tert-butyl 2-({[(tert-butoxy)carbonyl] ({2-[4-(methoxycarbonyl)-1,3-thiazol-2-yl] ethyl})amino}methyl)-1H-1,3-benzodiazol-1-carboxylate (7) (1.12 g) in THF/water (50 ml/10 ml) was stirred at room temperature for 16 hours. The reaction mixture acidified to pH ~1-2 with saturated KHSO4 solution and then evaporated in vacuo. The resulting crude residue was triturated in DCM/IPS, then MeOH/EtOAc to give the title compound (1.2 g, 50% pure) as an off-white solid.

1H NMR (Methanol ₄ , 500 MHz): δ [ppm] = 8.39 (s, 1H), 7.96-7.91 (m, 2H), 7.71-7.76 ( m, 2H), 5.07 (s, 2H), 3.84 (t, J=6.3 Hz, 2H), 3.64 (t, J=6.3 Hz, 2H).

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HPLC-MS (method A): [m/z]: 303.2 [M+H]+.

2-{2-[(1H-1,3-Benzodiazol-2-ylmethyl)amino]ethyl}-N-{[2-(morpholin-4-yl)phenyl]methyl}-1,3-thiazole4-carboxamide ( illustrative example No. 63)

Similar to general procedure 6, 2-{2-[(1H-1,3-Benzodiazol-2-ylmethyl)amino]ethyl}-1,3-thiazole-4carboxylic acid (80) (200 mg, 0.66 mmol, 50 % purity), 1-[2-(morpholin-4-yl)phenyl]methanamine (127 mg, 0.66 mmol), DIPEA (0.35 ml, 1.98 mmol) and HATU (377 mg, 0.99 mmol) in DMF (5 mL) gave the title compound (37 mg, 12%) as a yellow solid after purification by basic prep-HPLC.

¹ H NMR (Methanol-d4, 500 MHz): δ [ppm] = 8.06 (s, 1H), 7.50 (dd, J=5.9, 3.2 Hz, 2H), 7 .27 (t, J=8.0 Hz, 2H), 7.24-7.18 (m, 2H), 7.16 (d, J=7.6 Hz, 1H), 7.06 (t, J=7.6Hz, 1H), 4.67(s, 2H), 4.05(s, 2H), 3.88-3.80(m, 4H), 3.23(t, J=6 .8 Hz, 2H), 3.07 (t, J=6.8 Hz, 2H), 2.90-2.84 (m, 4H).

HPLC-MS (method B): [m/z]: 477.2 [M+H] ⁺ .

2-{2-[(1H-1,3-Benzodiazol-2-ylmethyl)amino]ethyl}-N-[(1-methyl-1H-imidazol-2-yl)methyl]-1,3thiazol-4- carboxamide (illustrative example No. 64)

Similar to general procedure 6, 2-{2-[(1H-1,3-Benzodiazol-2-ylmethyl)amino]ethyl}-1,3-thiazole-4carboxylic acid (80) (200 mg, 0.33 mmol, 50 % purity), 1-(1-methyl-1H-imidazol-2-yl)methanamine (36 mg, 0.33 mmol), DIPEA (230 µl, 1.32 mmol) and HATU (189 mg, 0.496 mmol) in DCM (5 ml) and DMF (1 ml) gave the title compound (15 mg, 12%) after purification by basic prep-HPLC.

1H NMR (Methanol-d ₄ , 500 MHz): δ [ppm] = 8.09 (s, 1H), 7.53 (dd, J=5.9, 3.2 Hz, 2H), 7 .26-7.20 (m, 2H), 7.03 (d, J=1.1 Hz, 1H), 6.89 (d, J=1.2 Hz, 1H), 4.66 (s, 2H), 4.08 (s, 2H), 3.73 (s, 3H), 3.25 (t, J=6.7 Hz, 2H), 3.11 (t, J=6.7 Hz, 2H).

HPLC-MS (method B): [m/z]: 396.2 [M+H] ⁺ .

2-{2-[(1H-1,3-Benzodiazol-2-ylmethyl)amino]ethyl}-N-[(2-fluorophenyl)methyl]-1,3-thiazole-4carboxamide (illustrative example No. 65)

Similar to general procedure 6, 2-{2-[(1H-1,3-benzodiazol-2-ylmethyl)amino]ethyl}-1,3-thiazole-4carboxylic acid (80) (200 mg, 0.53 mmol, 50 % purity), 1-(2-fluorophenyl)methanamine (66 mg, 0.53 mmol), DIPEA (369 μl, 2.12 mmol) and HATU (302 mg, 0.79 mmol) in DMF (2 ml) gave the title compound (13 mg, 6%) as a white solid after purification by basic prep-HPLC.

¹ H NMR (Methanol-d4, 500 MHz): δ [ppm] = 8.09 (s, 1H), 7.53 (dd, J=5.9, 3.2 Hz, 2H), 7 .40-7.35 (m, 1H), 7.32-7.26 (m, 1H), 7.25-7.19 (m, 2H), 7.15-7.05 (m, 2H) , 4.64 (s, 2H), 4.08 (s, 2H), 3.26 (t, J=6.7 Hz, 2H), 3.11 (t, J=6.7 Hz, 2H) .

HPLC-MS (Method D): [m/z]: 410.2 [M+H] ⁺ .

The above general scheme 4:

2-(2-{[(tert-butoxy)carbonyl]amino}ethyl)-1,3-thiazole-4-carboxylic acid (87)

Similar to general procedure 5, ethyl 2-(2-{[(tert-butoxy)carbonyl]amino}ethyl)-1,3-thiazole-4carboxylate (1) (8 g, 26.63 mmol) and LiOH (1.91 g, 79.90 mmol) in THF/water (200 ml/70 ml) at room temperature for 20 hours gave the title compound (10.16 g, 99.7%) as a yellow oil.

¹ H NMR (DMSO-d ₆ , 500 MHz): δ [ppm] = 8.32 (s, 1H), 6.99 (s, 1H), 3.28 (t, J=6.9 Hz, 2H overlap

- 120 039916 with solvent signals), 3.10 (t, J=6.9 Hz, 2H), 1.36 (s, 9H).

1H NMR (Methanol-d ₄ , 500 MHz): δ [ppm] = 8.24 (s, 1H), 3.46 (t, J=6.6 Hz, 2H), 3.19 (t , J=6.6 Hz, 2H),

1.41 (s, 9H).

HPLC-MS (method A): [m/z]: 294.9 [M+H]+.

2-(2-{[(tert-Butoxy)carbonyl]amino}ethyl)-5-methyl-1,3-thiazole-4-carboxylic acid (88) o χ-Ό A _NHBoc

Similar to general procedure 5, methyl 2-(2-{[(tert-butoxy)carbonyl]amino}ethyl)-5-methyl-1,3thiazole-4-carboxylate (3) (769 mg, 2.56 mmol) and LiOH (310 mg, 13 mmol) in THF/water (20 ml/20 ml) gave the title compound (681 mg, 88%) as a yellow oil.

1H NMR (DMSO-d ₆ , 250 MHz): δ [ppm] = 12.76 (s, 1H), 6.99 (t, J=5.6 Hz, 1H), 3.30-3 .19 (m, 2H), 3.01 (t, J=6.9 Hz, 2H), 2.65 (s, 3H), 1.37 (s, 9H).

HPLC-MS (method A): [m/z]: 301.05 [M+H] ⁺ .

2-(3-{[(tert-Butoxy)carbonyl]amino}propyl)-1,3-thiazole-4-carboxylic acid (89) o

NHBoc

Similar to general procedure 5, ethyl 2-(3-{[(tert-butoxy)carbonyl]amino}propyl)-1,3-thiazole-4carboxylate (4) (726 mg, 2.31 mmol) and LiOH (166 mg, 6.93 mmol) in THF (12 ml) and water (4 ml) gave the crude title compound (791 mg) as a yellow oil.

1H NMR (DMSO-d ₆ , 500 MHz): δ [ppm] = 8.13 (s, 1H), 6.91 (t, J=5.0 Hz, 1H), 3.05-2 .93 (m, 4H), 1.83 (n, J=7.2 Hz, 2H), 1.38 (s, 9H).

HPLC-MS (method A): [m/z]: 285 [M+H] ⁺ .

tert-Butyl N-[2-(4-{[(3-fluoropyridin-2-yl)methyl]carbamoyl}-1,3-thiazol-2-yl)ethyl]carbamate (90) F O

NHBoc

Similar to general procedure 6, (3-fluoropyridin-2-yl)methanamine dihydrochloride (A2) (8.03 g, 40.35 mmol), 2-(2-{[(tert-butoxy)carbonyl]amino}ethyl)- 1,3-thiazole-4-carboxylic acid (87) (10.2 g, 26.9 mmol), DIPEA (28.1 ml, 161.4 mmol) and HATU (12.3 g, 32.3 mmol) in THF (300 ml) at room temperature for 2 h gave the title compound (13.27 g) as an orange oil after purification by flash column chromatography (eluting with a 20-100% EtOAc/heptane gradient).

1H NMR (DMSO-d ₆ , 500 MHz): δ [ppm] = 8.70 (t, J=5.6 Hz, 1H), 8.39 (d, J=4.6 Hz, 1H ), 8.15 (s, 1H), 7.73-7.67 (m, 1H), 7.40 (dt, J=8.5, 4.4 Hz, 1H), 7.04 (s, 1H), 4.65 (d, J=5.6 Hz, 2H), 3.31 (t, J=6.8 Hz, 2H, overlap with solvent signals), 3.13 (t, J=6, 8 Hz, 2H), 1.36 (s, 9H).

HPLC-MS (method A): [m/z]: 381 [M+H] ⁺ .

tert-Butyl N-{2-[4-(benzylcarbamoyl)-1,3-thiazol-2-yl]ethyl}carbamate (91)

Similar to general procedure 6, 2-(2-{[(tert-butoxy)carbonyl]amino}ethyl)-1,3-thiazole-4carboxylic acid (87) (4.09 g, 15.0 mmol), benzylamine (1 .8 ml, 16.5 mmol), DIPEA (7.9 ml, 45.1 mmol) and HATU (8.570 g, 22.5 mmol) in DCM (205 ml) gave the title compound (3.38 g, 56%, 90% pure) as a yellow oil after purification by flash column chromatography (elution with 20-100% EtOAc/heptane gradient).

1H NMR (CDCl3, 500 MHz): δ [ppm] = 8.03 (s, 1H), 7.61 (s, 1H), 7.39-7.32 (m, 4H), 7, 31-7.27 (m, 1H), 4.64 (d, J=6.1 Hz, 2H), 3.54 (d, J=6.4 Hz, 2H), 3.17 (t, J = 6.4 Hz, 2H), 1.42 (s, 9H).

HPLC-MS (Method E): [m/z]: 384 [M+Na]+.

tert-Butyl N-(2-{4-[(pyridin-2-ylmethyl)carbamoyl]-1,3-thiazol-2-yl}ethyl)carbamate (92)

Similar to general procedure 6, 2-(2-{[(tert-butoxy)carbonyl]amino}ethyl)-1,3-thiazole-4carboxylic acid (87) (0.500 g, 1.836 mmol), 1-(pyridine-2- yl)methanamine (0.199 g, 1.836 mmol), HATU (1.047 g, 2.754 mmol) and DIPEA (0.959 ml, 5.508 mmol) in DCM (25 ml) gave the title compound (0.905 g, quant) as a yellow oil after purification by flash column chromatography (using a gradient of 20% heptane:80% ethyl acetate to 100% ethyl acetate).

HPLC-MS (method A): [m/z]: 363.05 [M+H] ⁺ .

tert-Butyl N-(2-{4-[(pyride azin-3-yl-methyl)carbamoyl]-1,3-thiazol-2-yl}ethyl)carbamate (93)

- 121 039916

Similar to general procedure 6, 2-(2-{[(tert-butoxy)carbonyl]amino}ethyl)-1,3-thiazole-4carboxylic acid (87) (0.7 g, 2.57 mmol), pyridazine-3 -ylmethanamine (0.42 g, 3.86 mmol), DIPEA (2.24 ml, 12.85 mmol) and HATU (1.47 g, 3.86 mmol) in DMF (15 ml) gave the title compound (0.919 g, 98%) as a brown oil after purification by flash column chromatography (eluting with a 0-5% MeOH-DCM gradient).

1H NMR (CDCl ₃ , 500 MHz): δ [ppm] = 9.21 (d, J=4.0 Hz, 1H), 8.41-8.32 (bdm, 1H), 8 .03 (s, 1H), 7.917.85 (br. m, 1H), 7.77-7.69 (b. m, 1H), 5.01 (d, J=5.7 Hz, 2H), 4.96 (brs, 1H), 3.65-3.46 (m, 2H), 3.20 (t, J=6.2 Hz, 2H), 1.42 (s, 9H).

HPLC-MS (method A): [m/z]: 364.05 [M+H] ⁺ .

tert-Butyl N-[2-(4-{[(6-methylpyridazin-3-yl)methyl]carbamoyl}-1,3-thiazol-2-yl)ethyl]carbamate (94)

Similarly, using the general procedure 6, 2-(2-{[(tert-butoxy)carbonyl]amino}ethyl)1,3-thiazole-4-carboxylic acid (87) (1.3 g, 4.15 mmol) , (6-methylpyridazin-3-yl)methanamine hydrochloride (0.8 g, 5.01 mmol), DIPEA (2.89 ml, 16.61 mmol) and HATU (1.9o g, 5.01 mmol) in THF (35 mL) and DMF (5 mL) gave the title compound (0.878 g, 45%) as a colorless oil after purification by flash column chromatography (kp-NH, eluting with a 0-15% MeOH/EtOAc gradient) followed by a second purification by flash column chromatography (kp-NH, eluting with a 70-100% EtOAc/heptane gradient).

1H NMR (DMSO-d6, 500 MHz): δ [ppm] = 9.04 (t, J=6.0 Hz, 1H), 8.17 (s, 1H), 7.50 (q, J=8.6Hz, 2H), 7.05(s, 1H), 4.71(d, J=6.1Hz, 2H), 3.13(t, J=6.8Hz, 2H) , 2.59 (s, 3H), 1.36 (s, 9H).

HPLC-MS (Method A): [m/z]: 378.05 [M+H] ⁺ .

tert-Butyl N-(2-{4-[(pyrimidin-2-ylmethyl)carbamoyl]-1,3-thiazol-2-yl}ethyl)carbamate (95)

Similar to general procedure 6, 2-(2-{[(tert-butoxy)carbonyl]amino}ethyl)-1,3-thiazole-4carboxylic acid (87) (0.7 g, 2.57 mmol), pyrimidine-2 -ylmethanamine (0.42 g, 3.86 mmol), DIPEA (2.24 ml, 12.85 mmol) and HATU (1.47 g, 3.86 mmol) in DMF (15 ml) gave the title compound (0.545 g, 58%) as a pale yellow solid after purification by flash column chromatography (eluting with a 0-5% MeOH/DCM gradient).

1H NMR (CDCl ₃ , 250 MHz): δ [ppm] = 8.83 (s, 1H), 8.81 (s, 1H), 8.46 (br s, 1H), 8.06 (s, 1H), 7.32 (pseudo t, J=4.8 Hz, 1H), 5.01 (broad s, 1H), 4.97 (d, J=5.3 Hz, 2H), 3.67-3.56 (m, 2H), 3.26 (t, J=6.4 Hz, 2H), 1.47 (s, 9H).

HPLC-MS (method A): [m/z]: 364.05 [M+H] ⁺ .

tert-Butyl (96)

N-[2-(4-{[(5-methylpyrimidin-2-yl)methyl]carbamoyl}-1,3-thiazol-2-yl)ethyl]carbamate

Similar to general procedure 6, 2-(2-{[(tert-butoxy)carbonyl]amino}ethyl)-1,3-thiazole-4 carboxylic acid (87) (500 mg, 1.6 mmol, 87% pure), (5-methylpyrimidin-2-yl)methanamine (295 mg, 2.4 mmol), DIPEA (1.39 ml, 7.99 mmol) and HATU (911 mg, 2.4 mmol) in THF (15 ml) and DMF (3 mL) gave the crude title compound (600 mg, 85%, 85% pure) as a yellow oil after purification by flash chromatography (eluting with a 0-80% EtOAc/heptane gradient).

HPLC-MS (Method A): [m/z]: 378.10 [M+H]+.

tert-Butyl yl]ethyl}carbamate (97)

N-{2-[4-({5H,6H,7H-cyclopenta[b]pyridin-7-yl}carbamoyl)-1,3-thiazole-2-

Similar to general procedure 6, 2-(2-{[(tert-butoxy)carbonyl]amino}ethyl)-1,3-thiazole-4carboxylic acid (87) (1.2 g, 4.41 mmol), 5H,6H ,7H-cyclopenta[b]pyridine-7-amine hydrochloride (1.13 g, 6.61 mmol), DIPEA (2.3 ml, 13.22 mmol) and HATU (2.51 g, 6.61 mmol) in DMF (24 ml) gave the indicated

- 122 039916 title compound (1.53 g, 85%) as a pale pink powder after purification by flash column chromatography (elution with a 0-5% MeOH/DCM gradient).

HPLC-MS (Method A): [m/z]: 389.15 [M+H]+.

tert-Butyl N-[2-(4-{[(3-methoxypyridin-2-yl)methyl]carbamoyl}-1,3-thiazol-2-yl)ethyl]carbamate (98) OMe η NHBoc

Similar to general procedure 6, 2-(2-{[(tert-butoxy)carbonyl]amino}ethyl)-1,3-thiazole-4 carboxylic acid (87) (250 mg, 0.918 mmol), (3-methoxypyridine-2 -yl)methanamine dihydrochloride (213 mg, 1.01 mmol), DIPEA (0.80 ml, 4.59 mmol) and HATU (524 mg, 1.38 mmol) in DCM (15 ml) gave the crude title compound (417 mg) as a yellow oil after purification by flash column chromatography (eluting with a 30-100% EtOAc/heptane gradient).

1H NMR (CDCl ₃ , 500 MHz): δ [ppm] = 8.68 (s, 1H), 8.21 (d, J=4.7 Hz, 1H), 8.02 (s, 1H ), 7.21 (dd, J=8.2, 4.7 Hz, 1H), 7.16 (d, J=8.3 Hz, 1H), 5.02 (s, 1H), 4.76 (d, J=4.7 Hz, 2H), 3.89 (s, 3H), 3.63 (d, J=6.0 Hz, 2H), 3.23 (t, J=6.0 Hz , 2H), 1.44 (s, 9H).

HPLC-MS (method A): [m/z]: 393.40 [M+H] ⁺ .

tert-Butyl N-[2-(4-{[(3-methoxypyridin-2-yl)methyl]carbamoyl}-1,3-thiazol-2-yl)ethyl]carbamate (99) o

Similar to general procedure 6, 2-(2-{[(tert-butoxy)carbonyl]amino}ethyl)-1,3-thiazole-4-carboxylic acid (87) (250 mg, 0.918 mmol), 1-(1H-benzimidazole- 2-yl)methanamine (149 mg, 1.01 mmol), DIPEA (0.48 ml, 2.75 mmol) and HATU (524 mg, 1.38 mmol) in DCM (15 ml) gave the crude title compound (0.709 mg, quantitative yield, 81% purity) as a yellow oil after purification by flash column chromatography (eluting with a 0-10% MeOH/DCM gradient).

HPLC-MS (method A): [m/z]: 402 [M+H] ⁺ .

tert-Butyl N-[2-(4-{[(3-fluoropyridin-2-yl)methyl]carbamoyl}-5-methyl-1,3-thiazol-2yl)ethyl]carbamate (100)

Similar to general procedure 6, 2-(2-{[(tert-butoxy)carbonyl]amino}ethyl)-5-methyl-1,3-thiazole-4-carboxylic acid (88) (680 mg, 2.37 mmol), ( 3-fluoropyridin-2-yl)methanamine dihydrochloride (A2) (615 mg, 3.09 mmol), TEA (1.16 ml, 8.0 mmol) and HATU (1350 mg, 3.56 mmol) in DCM (30 ml) gave the title compound (824 mg, 85%) as a yellow oil.

¹ H NMR (CDCl ₃ , 500 MHz): δ [ppm] = 8.45 (m, 2H), 7.42 (ddd, J=9.4, 8.3, 1.2 Hz, 1H ), 7.30-7.24 (m, 1H), 4.99 (s, 1H), 4.83 (dd, J=5.2, 1.5 Hz, 2H), 3.59 (d, J=6.0 Hz, 2H), 3.13 (t, J=6.0

Hz, 2H), 2.83 (s, 3H), 1.46 (s, 9H).

HPLC-MS (method A): [m/z]: 395.15 [M+H] ⁺ .

tert-Butyl N-(2-{5-methyl-4-[(pyrimidin-2-ylmethyl)carbamoyl]-1,3-thiazol-2-yl}ethyl)carbamate (101) o

IL ^,ν n L > \ /% ^NHBoc

Similar to general procedure 6, 2-(2-{[(tert-butoxy)carbonyl]amino}ethyl)-5-methyl-1,3-thiazole-4-carboxylic acid (88) (0.315 g, 0.912 mmol), 1-( pyrimidin-2-yl)methanamine (0.119 g, 1.094 mmol), THF (7 mL), DMF (1 mL), DIPEA (0.318 mL, 1.824 mmol) and HATU (0.416 g, 1.094 mmol) gave the title compound ( 0.134 g, 27%) as a colorless oil after purification by flash column chromatography (gradient 30-100% EtOAc in heptane).

HPLC-MS (method A): [m/z]: 378.10 [M+H] ⁺ .

tert-Butyl N-[3-(4-{[(3-fluoropyridin-2-yl)methyl]carbamoyl}-1,3-thiazol-2-yl)propyl]carbamate (102)

F O

Un ⁿ

NHBoc

Similar to general procedure 6, 2-(3-{[(tert-butoxy)carbonyl]amino}propyl)-1,3-thiazole-4carboxylic acid (89) (661 mg, 2.31 mmol), (3-fluoropyridine- 2-yl)methanamine dihydrochloride (A2) (689

- 123039916 mg, 3.46 mmol), DIPEA (2.41 ml, 13.85 mmol) and HATU (1053 mg, 2.77 mmol) in DMF (4 ml) and THF (4 ml) gave the title compound (914 mg, 93%, 93% pure) as a yellow oil after purification by flash column chromatography (elution with 20-100% EtOAc/heptane gradient).

HPLC-MS (method A): [m/z]: 395.05 [M+H]+.

2-(2-Aminoethyl)-N-[(3-fluoropyridin-2-yl)methyl]-1,3-thiazole-4-carboxamide dihydrochloride (103)

Similar to general procedure 2, 12M HC (35.3 ml) and tert-butyl N-[2-(4-{[(3-fluoropyridin-2yl)methyl]carbamoyl}-1,3-thiazol-2-yl)ethyl ]carbamate (90) (13.3 g, 28.25 mmol) in MeOH (250 ml) was stirred at 50° C. for 3 h. The resulting mixture was evaporated in vacuo to give the title compound (12.8 g) in form of an off-white solid.

1H NMR (DMSO-a ₆ , 500 MHz): δ [ppm] = 8.98 (s, 1H), 8.39 (d, J=4.7 Hz, 1H), 8.21 (s , 1H), 8.17 (s, 3H), 7.72 (t, J=9.3 Hz, 1H), 7.42 (dt, J=8.5, 4.4 Hz, 1H), 4 .67 (d, J=5.8 Hz, 2H), 3.38 (t, J=6.5 Hz, 2H), 3.30-3.25 (m, 2H).

HPLC-MS (method A): [m/z]: 280.9 [M+H] ⁺ .

2-(2-Aminoethyl)-N-benzyl-1,3-thiazole-4-carboxamide hydrochloride (104)

Similar to general procedure 2, 12M HC (2.5 ml) and tert-butyl N-{2-[4-(benzylcarbamoyl)-1,3thiazol-2-yl]ethyl}carbamate (91) (456 mg, 1.29 mmol) in MeOH (4.5 ml) at room temperature for 4 hours gave the title compound (336 mg, 100%) as a beige solid. The resulting product was used in subsequent reactions without purification.

HPLC-MS (Method E): [m/z]: 261.95 [M+H] ⁺ .

2-(2-Aminoethyl)-N-(pyridin-2-ylmethyl)-1,3-thiazole-4-carboxamide dihydrochloride (105)

Similar to general procedure 2, tert-Butyl N-(2-{4-[(pyridin-2-ylmethyl)carbαmoyl]-1,3-thiazol-2yl}ethyl)carbamate (92) (0.905 g, 2.50 mmol) , 12M HC (4.852 mL, 58.23 mmol) in MeOH (9 mL) gave the title compound (0.840 g, quant) as a white solid.

HPLC-MS (method A): [m/z]: 262.95 [M+H] ⁺ .

2-(2-Aminoethyl)-N-(pyridazin-3-ylmethyl)-1,3-thiazole-4-carboxamide dihydrochloride (106)

Similar to general procedure 2, t-butyl N-(2-{4-[(pyridazin-3-ylmethyl)carbamoyl]-1,3-thiazol2-yl}ethyl)carbamate (93) (0.919 g, 2.53 mmol) and 12M HC (4.22 ml) in MeOH (15 ml) at room temperature for 16 hours gave the title compound (0.840 g, 97%) as a brown residue.

1H NMR (Deuterium oxide, 500 MHz): δ [ppm] = 9.17 (dd, J=4.9, 1.5 Hz, 1H), 8.12 (s, 1H), 8.03 (dd, J=8.6, 1.5 Hz, 1H), 7.99 (dd, J=8.6, 4.9 Hz, 1H), 4.86 (s, 2H), 3.45- 3.39 (m, 2H), 3.48-3.43 (m, 2H).

HPLC-MS (Method A): [m/z]: 263.95 [M+H]+.

2-(2-Aminoethyl)-N-[(6-methylpyridazin-3-yl)methyl]-1,3-thiazole-4-carboxamide dihydrochloride (107)

Similar to general procedure 2, tert-butyl N-[2-(4-{[(6-methylpyridazin-3-yl)methyl]carbamoyl}1,3-thiazol-2-yl)ethyl]carbamate (94) (878 mg , 1.86 mmol) and 12M HC (3.10 ml) in MeOH (15 ml) at room temperature for 24 hours gave the title compound (764 mg, quant) as an off-white solid. The resulting product was used in subsequent reactions without purification.

1H NMR (MeOD, 500 MHz): δ [ppm] = 8.49 (d, J=8.9 Hz, 1H), 8.41 (d, J=8.9 Hz, 1H), 8 .26 (s, 1H), 4.97 (s, 2H), 3.53-3.47 (m, 4H), 2.91 (s, 3H).

HPLC-MS (method A): [m/z]: 277.95 [M+H] ⁺ .

2-(2-Aminoethyl)-N-(pyrimidin-2-ylmethyl)-1,3-thiazole-4-carboxamide dihydrochloride (108)

- 124 039916

Similar to general procedure 2, tert-butyl X-(2-{4-[(pyrimidin-2-ylmethyl)carbamoyl]-1,3-thiazol2-yl}ethyl)carbamate (95) (0.545 g, 1.499 mmol) and 12M HC (4.22 ml) in MeOH (15 ml) at room temperature over 16 hours gave the title compound (0.530 g, quant) as a pale yellow foam.

1H NMR (Deuterium oxide, 500 MHz): δ [ppm] = 8.73 (d, J=5.1 Hz, 2H), 8.16 (s, 1H), 7.47 (pseudo t, J=5.1 Hz, 1H), 4.80 (s, 2H), 3.51-3.46 (m, 2H), 3.45-3.41 (m, 2H).

HPLC-MS (ESI+): [m/z]: 263.95 [M+H] ⁺ .

as free base (METCR1673 Generic 2 min) 2-(2-aminoethyl)-N-[(5-methylpyrimidin2-yl)methyl]-1,3-thiazole-4-carboxamide (109)

Similar to general procedure 2, tert-butyl N-[2-(4-{[(5-methylpyrimidin-2-yl)methyl]carbamoyl}1,3-thiazol-2-yl)ethyl]carbamate (96) (600 mg , 1.59 mmol) and 12 M HC (2.65 mL) in MeOH (10 mL) gave the title compound as the free base (283 mg, 44%) as a white solid after purification by flash column chromatography (eluting with a gradient of 0-10% 7 M NH ₃ in MeOH/DCM).

1H NMR (DMSO-d ₆ , 500 MHz): δ [ppm] = 8.77-8.70 (m, 1H), 8.62 (s, 2H), 8.14 (s, 1H) , 4.64 (d, J=4.0 Hz, 2H), 3.13 (t, J=6.6 Hz, 2H), 3.02 (t, J=6.6 Hz, 2H), 2 .26 (s, 3H).

HPLC-MS (method A): [m/z]: 278.2 [M+H] ⁺ .

2-(2-Aminoethyl)-N-{5Н,6Н,7Н-cyclopenta[b]pyridin-7-yl}-1,3-thiαzol-4-carboxamide dihydrochloride (110)

Similar to general procedure 2, a 4M solution of HC in dioxane (14.45 ml, 57.8 mmol) was added to an ice-cold solution of tert-butyl N-{2-[4-({5H,6H,7H-cyclopenta[b]pyridine- 7-yl}carbamoyl)-1,3-thiαzol-2yl]ethyl}carbamate (97) (1.53 g, 3.94 mmol) in MeOH (5 ml). The resulting mixture was stirred at room temperature for 2 hours. The resulting title compound (1.32 g, 93%) was separated by filtration after precipitation from diethyl ether (5 ml).

HPLC-MS (method A): [m/z]: 289.05 [M+H] ⁺ .

2-(2-Aminoethyl)-N-[(3-methoxypyridin-2-yl)methyl]-1,3-thiazole-4-carboxamide (111)

Similar to general procedure 2, 12M HC (2.5 ml) and crude tert-butyl X-[2-(4-{[(3-methoxypyridin2-yl)methyl]carbamoyl}-1,3-thiazol-2-yl) ethyl]carbamate (98) (417 mg) in MeOH (5 mL) at room temperature for 2 h gave the title compound (125 mg) as a white solid after purification by flash column chromatography (kp-NH, eluting in a 0-10% MeOH/DCM gradient).

¹ H NMR (DMSO-66, 500 MHz): δ [ppm] = 8.59 (s, 1H), 8.15-8.11 (m, 2H), 7.44 (d, J= 8.3 Hz, 1H), 7.33 (dd, J=8.3, 4.7 Hz, 1H), 4.57 (d, J=5.2 Hz, 2H), 3.88 (s, 3H), 3.08 (t, J=6.6 Hz, 2H), 2.94 (t, J=6.6 Hz, 2H), 2.69 (s, 2H).

HPLC-MS (Method A): [m/z]: 292.95 [M+H]+.

2-(2-Aminoethyl)-N-( 1 H-1,3-benzodiazol-2-ylmethyl)-1,3-thiazol-4-carboxamide (112)

Similar to general procedure 2, 12M HC (2.5 ml) and crude t-butyl ^(2-{4-[(N-1,3-benzodiazol2-ylmethyl)carbamoyl]-1,3-thiazol-2-yl} ethyl) carbamate (99) (709 mg, 1.43 mmol, 81% purity) in MeOH (5 ml) at room temperature for 2 h gave the title compound (111 mg, 25%) as a brown solid after purification by flash column chromatography (kp-NH, eluting with a 0-10% MeOH/DCM gradient).

1H NMR (DMSO-d6, 500 MHz): δ [ppm] = 8.88 (s, 1H), 8.17 (s, 1H), 7.49 (s, 2H), 7.13 ( dd, J=6.0, 3.1 Hz,

- 125 039916

2H), 4.68 (d, J=6.0 Hz, 2H), 3.08 (t, J=6.6 Hz, 2H), 2.95 (t, J=6.6 Hz, 2H) .

HPLC-MS (Method A): [m/z]: 301.95 [M+H]+.

2-(2-Aminoethyl)-N-[(3-fluoropyridin-2-yl)methyl]-5-methyl-1,3-thiazole-4-carboxamide dihydrochloride (113)

Similar to general procedure 2, tert-butyl N-[2-(4-{[(3-fluoropyridin-2-yl)methyl]carbαmoyl}-5methyl-1,3-thiazol-2-yl)ethyl]carbamate (100) (823 mg, 2.09 mmol) and 12M HC (3 ml) in MeOH (30 ml) gave the title compound (794 mg, quant.) as a brownish solid.

1H NMR (DMSO-d6, 250 MHz): δ [ppm] = 8.87 (t, J=5.9 Hz, 1H), 8.40 (dt, J=4.4, 1.3 Hz, 1H), 8.13 (s, 3H), 7.73 (ddd, J=9.9, 8.3, 1.2 Hz, 1H), 7.43 (dt, J=8.5, 4.5 Hz, 1H), 4.69-4.60 (m, 2H), 3.32-3.19 (m, 4H), 2.71 (s, 3H).

HPLC-MS (Method A): [m/z]: 295.05 [M+H]+.

2-(2-Aminoethyl)-5-methyl-N-(pyrimidin-2-ylmethyl)-1,3-thiazole-4-carboxamide dihydrochloride (114)

2HCI

Similar to general procedure 2, tert-butyl N-(2-{5-methyl-4-[(pyrimidin-2-ylmethyl)carbamoyl]1,3-thiazol-2-yl}ethyl)carbamate (101) (0.482 g, 0.795 mmol), MeOH (6 ml) and 12M HC (1.325 ml, 15.90 mmol) gave the title compound (0.420 g, 99%) as a yellow solid.

HPLC-MS (method A): [m/z]: 277.95 [M+H] ⁺ .

2-(3-aminopropyl)-N-[(3-fluoropyridin-2-yl)methyl]-1,3-thiazole-4-carboxαmid dihydrochloride (115) °

NH, 2HCI ²

Similar to general procedure 2, 4M solution of HC in dioxane (2.89 ml, 11.55 mol) and tert-butyl N-[3(4-{[(3-fluoropyridin-2-yl)methyl]carbamoyl}-1, 3-thiazol-2-yl)propyl]carbamate (102) (0.91 g, 2.31 mmol) in dioxane (6 ml) and MeOH (2 ml) gave the title compound (1.13 g, 85% , 64% pure) as a pale orange oil. The resulting compound was used in the next step without purification.

HPLC-MS (Method A): [m/z]: 295.00 [M+H] ⁺ .

2-{2-[(1H-1,3-Benzodiazol-2-ylmethyl)amino]ethyl}-N-[(3-methoxypyridin-2-yl)methyl]-1,3-thiazole-4carboxamide (Illustrative Example No. 59)

Similarly, using the general procedure 3, 2-(2-Aminoethyl)-N-[(3-methoxypyridin-2yl)methyl]-1,3-thiazole-4-carboxamide (111) (90 mg, 0.308 mmol) and 1H -benzimidazole-2-carbaldehyde (49 mg, 0.339 mmol) in DCE (9 ml) at room temperature for 2 h, followed by addition of NaBH(OAc) ₃ (91 mg, 0.431 mmol) gave the title compound (50 mg, 38%) as a yellow solid after purification by basic prep-HPLC.

1H NMR (DMSO-d6, 500 MHz): δ [ppm] = 12.18 (s, 1H), 8.58 (t, J=5.2 Hz, 1H), 8.14 (s, 1H), 8.09-8.06 (m, 1H), 7.59-7.40 (m, 3H), 7.31 (dd, J=8.3, 4.7 Hz, 1H), 7 .12 (d, J=3.5 Hz, 2H), 4.56 (d, J=5.2 Hz, 2H), 3.97 (s, 2H), 3.87 (s, 3H), 3 .20 (t, J=6.8 Hz, 2H), 2.98 (t, J=6.8 Hz, 2H).

HPLC-MS (method B): [m/z]: 423.2 [M+H] ⁺ .

N-(1H-1,3-Benzodiazol-2-ylmethyl)-2-{2-[(1H-1,3-benzodiazol-2-ylmethyl)amino]ethyl}-1,3-thiazole-4carboxamide (illustrative example No. 60) about

Similarly, using General Procedure 3, 2-(2-aminoethyl)-N-(1H-1,3-benzodiazol-2ylmethyl)-1,3-thiazole-4-carboxamide (112) (111 mg, 0.368 mmol) and ¹ H-benzimidazole-2-carbaldehyde (59 mg, 0.405 mmol) in DCE (12 ml) at room temperature for 2 hours, followed by addition

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NaBH(OAc) ₃ (109 mg, 0.516 mmol) gave the title compound (15 mg, 9%) as a yellow solid after purification by basic prep-HPLC.

1H NMR (DMSO-b ₆ , 500 MHz): δ [ppm] = 12.23 (s, 2H), 8.89 (t, J=5.9 Hz, 1H), 8.18 (s , 1H), 7.49 (s, 4H),

7.13 (tt, J=7.0, 3.5 Hz, 4H), 4.69 (d, J=5.9 Hz, 2H), 3.98 (s, 2H), 3.21 (t , J=6.8 Hz, 2H), 3.00 (t, J=6.8 Hz, 2H).

HPLC-MS (method B): [m/z]: 430.3 [M-H] ⁺ .

2-(2-{[(6-Fluoro-1H-1,3-benzodiazol-2-yl)methyl]amino}ethyl)-N-[(3-fluoropyridin-2-yl)methyl]-1,3thiazol- 4-carboxamide trihydrochloride (illustrative example No. 114)

Similar to general procedure 3, 2-(2-Aminoethyl)-N-[(3-fluoropyridin-2-yl)methyl]-1,3-thiazole-4-carboxamide dihydrochloride (103) (200 mg, 0.49 mmol), 6 -fluoro-1H-benzimidazole-2-carbaldehyde (89 mg, 0.539 mmol) and DIPEA (0.342 ml, 1.961 mmol) in MeOH (10 ml) at room temperature for 24 h, followed by the addition of NaBH ₄ (28 mg, 0.735 mmol) gave the title compound (83 mg, free base) as a brown solid after purification by basic pre-HPLC. The resulting free base and 12M HC (1 ml) in MeOH (4 ml) were stirred at room temperature to give the title compound (111 mg, 42%) after the solvent was evaporated in vacuo.

¹ H NMR (Methanol-a ₄ , 500 MHz): δ [ppm] = 8.52 (s, 1H), 8.28 (s, 1H), 8.24 (s, 1H), 7, 86 (s, 1H), 7.78 (s, 1H), 7.53 (s, 1H), 7.34 (s, 1H), 4.97 (s, 2H), 4.92 (s, 2H ), 3.84 (s, 2H), 3.64 (t, J=6.1 Hz, 2H).

HPLC-MS (Method D): [m/z]: 429.1 [M+H] ⁺ .

2-{2-[(1H-1,3-Benzodiazol-5-ylmethyl)amino]ethyl}-N-[(3-fluoropyridin-2-yl)methyl]-1,3-thiazole-4carboxamide (Illustrative Example No. 139)

As in general procedure 3, 2-(2-Aminoethyl)-N-[(3-fluoropyridin-2-yl)methyl]-1,3-thiazole-4-carboxamide dihydrochloride (103) (400 mg, 0.64 mmol, 56% purity), 1H-1,3-benzodiazol-5-carbaldehyde (112 mg, 0.77 mmol) and DIPEA (0.56 ml, 3.19 mmol) in MeOH (10 ml) at room temperature for 18 h, followed by addition of NaBH ₄ (36 mg, 0.96 mmol) gave the title compound (207 mg, 75.9%) as a cream solid after purification by flash column chromatography (KP-NH, eluting with a 0- 20% MeOH/DCM).

1H NMR (Methanol-b ₄ , 500 MHz): δ [ppm] = 8.31-8.29 (m, 1H), 8.14 (s, 1H), 8.10 (s, 1H) , 7.64-7.53 (m, 3H), 7.37-7.33 (m, 1H), 7.29 (dd, J=8.3, 1.3 Hz, 1H), 4.79 (d, J=1.6 Hz, 2H), 3.96 (s, 2H), 3.28 (t, J=6.9 Hz, 2H), 3.09 (t, J=6.9 Hz , 2H).

HPLC-MS (Method C): [m/z]: 411.2 [M+H] ⁺ .

2-{2-[(1H-1,3-Benzodiazol-4-ylmethyl)amino]ethyl}-N-[(3-fluoropyridin-2-yl)methyl]-1,3-thiazole-4carboxamide (Illustrative Example No. 140)

Similarly, using the general procedure 3, 2-(2-α-minoethyl)-N-[(3-fluoropyridin-2yl)methyl]-1,3-thiazole-4-carboxamide dihydrochloride (103) (400 mg, 0.64 mmol , 56.3% pure), 1H-1,3-benzodiazol-4-carbaldehyde (112 mg, 0.77 mmol) and DIPEA (0.56 ml, 3.19 mmol) in MeOH (10 ml) at room temperature for 18 h followed by addition of NaBH ₄ (36 mg, 0.96 mmol) gave the title compound (255 mg, 96.4%) as an off-white solid after purification by flash column chromatography (KP-NH , eluting with a 0-10% MeOH/DCM gradient).

1H NMR (DMSO-a ₆ , 500 MHz): δ [ppm] = 12.41 (s, 1H), 8.65 (t, J=5.5 Hz, 1H), 8.39-8 .36 (m, 1H), 8.17 (brs, 1H), 8.12 (brs, 1H), 7.72-7.67 (m, 1H), 7.43-7.37 (m, 1H), 7.22-7.08 (m, 2H), 4.66 (dd, J=5.7, 1.4 Hz, 2H), 4.13 (s, 1H), 4, 04 (s, 1H), 3.18 (t, J=6.5 Hz, 2H), 2.97-2.87 (m, 2H).

HPLC-MS (Method C): [m/z]: 411.2 [M+H] ⁺ .

2-{3-[(1H-1,3-Benzodiazol-2-ylmethyl)amino]propyl}-N-[(3-fluoropyridin-2-yl)methyl]-1,3-thiazole-4carboxamide trihydrochloride (illustrative example No. 134)

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Similar to general procedure 3, 2-(3-Aminopropyl)-I-[(3-fluoropyridin-2-yl)methyl]-1,3-thiazole-4-carboxamide dihydrochloride (115) (150 mg, 0.261 mmol, 64% pure) , 1H-benzimidazole-2-carbaldehyde (46 mg, 0.314 mmol), DIPEA (0.18 ml, 1.05 mmol) in MeOH (2 ml) at room temperature for 16 h, followed by the addition of NaBH ₄ (15 mg , 0.39 mmol) gave the compound free base (53 mg, 48%) as a colorless oil after purification by basic prep-HPLC.

1H NMR (DMSO-J6, 250 MHz): δ [ppm] = 12.16 (s, 1H), 8.66 (t, J=5.5 Hz, 1H), 8.38 (dt, J=4.7, 1.4Hz, 1H), 8.13(s, 1H), 7.70(ddd, J=10.0, 8.4, 1.2Hz, 1H), 7.61 -7.27 (m, 2H), 7.11 (dd, J=6.0, 3.1 Hz, 2H), 4.65 (dd, J=5.5, 1.4 Hz, 2H), 3.91 (s, 2H), 3.09 (t, 2H), 2.64 (t, J=6.8 Hz, 2H), 1.92 (m, 2H).

HPLC-MS (Method D): [m/z]: 425.2 [M+H]+.

The resulting free base (35 mg, 0.082 mmol) and 12M HC (20 μl, 0.247 mmol) were stirred in MeOH (2 ml) at room temperature to give the title compound (44 mg, quant) as a white solid after evaporation solvent in a vacuum.

1H NMR (D ₂ O, 500 MHz): δ [ppm] = 8.50 (dd, J=5.4, 1.2 Hz, 1H), 8.19-8.12 (m, 2H ), 7.89-7.78 (m, 3H), 7.66 (dt, J=6.3, 3.3 Hz, 2H), 4.89 (s, 2H), 4.85 (d, J=1.3 Hz, 2H), 3.46-3.39(m, 2H), 3.26(t, J=7.2 Hz, 2H), 2.33(m, 2H).

HPLC-MS (Method D): [m/z]: 425.2 [M+H] ⁺ .

2-{2-[(1H-1,3-Benzodiazol-2-ylmethyl)amino]ethyl}-N-[(3-fluoro-pyridin-2-yl)methyl]-5-methyl-1,3thiazol-4-carboxamide (illustrative example #164)

Similarly, using the general procedure 3, 2-(2-aminoethyl)-N-[(3-fluoropyridin-2yl)methyl]-5-methyl-1,3-thiazole-4-carboxamide dihydrochloride (113) (274 mg, 0.75 mmol), 1H-1,3-benzodiazol-2-carbaldehyde (109 mg, 0.75 mmol), DIPEA (0.45 ml, 2.61 mmol) and anhydrous MgSO ₄ (200 mg) in MeOH (10 ml ) and DCM (10 ml) at room temperature for 20 hours, followed by the addition of NaBH ₄ (60 mg, 1.48 mmol) gave the title compound (140 mg, 44%) as a pale yellow solid after purification by method preparative HPLC.

1H NMR (DMSO-J6, 500 MHz): δ [ppm] = 12.17 (s, 1H), 8.59 (t, J=5.6 Hz, 1H), 8.36 (dt, J=4.7, 1.4Hz, 1H), 7.70(ddd, J=9.9, 8.4, 1.1Hz, 1H), 7.53(d, J=7.5Hz , 1H), 7.44 (d, J=7.5 Hz, 1H), 7.40 (dt, J=8.5, 4.4 Hz, 1H), 7.13 (n, J=6, 6 Hz, 2H), 4.65-4.59 (m, 2H), 3.96 (s, 2H), 3.10 (t, J=6.8 Hz, 2H), 2.94 (t, J=6.8 Hz, 2H), 2.68 (s, 3H).

HPLC-MS (method C): [m/z]: 425.2 [M+H] ⁺ .

General Procedure 7: 2-(2-{[2-(1H-1,3-Benzodiazol-2-yl)ethyl]amino}ethyl)-N-[(3-fluoropyridin-2yl)methyl]-1,3- thiazole-4-carboxamide (illustrative example No. 94) o

R2 _X JL m

R3 L --X N short

R4

2-(2-Aminoethyl)-N-[(3-fluoropyridin-2-yl)methyl]-1,3-thiazole-4-carboxamide dihydrochloride (103) (2.0 g, 3.96 mmol) was added to the solution 2-(2-chloroethyl)-1H-1,3-benzodiazol hydrochloride (1.12 g, 5.15 mmol) and DIPEA (10.6 ml, 59.45 mmol) in DMF (60 ml). The reaction mixture was stirred at 30°C for 6 days (the progress of the reaction was monitored by LCMS). The resulting mixture was evaporated in vacuo and the resulting residue was neutralized with saturated aqueous NaHCO ₃ . The aqueous layer was extracted with 4:1 CHS3/IPA (4x100 ml) and the combined organic layers were dried (MgSO ₄ ), filtered and evaporated in vacuo. The resulting crude residue was purified by flash column chromatography (kp-NH, eluting with a gradient of 60-100% EtOAc/heptane, then 0-20% MeOH/EtOAc) followed by neutral reverse phase column chromatography (eluting with a gradient of 0-60% MeCN /water) to give the title compound (0.173 g, 10%) as a yellow oil.

1H NMR (Methanol-a ₄ , 500 MHz): δ [ppm] = 8.31 (d, J=4.6 Hz, 1H), 8.02 (s, 1H), 7.57 (t , J=9.1 Hz, 1H), 7.45-7.40 (m, 2H), 7.36 (dd, J=8.6, 4.3 Hz, 1H), 7.17 (dd, J=6.0, 3.2 Hz, 2H), 4.68 (s, 2H), 3.26 (d, J=6.8 Hz, 2H), 3.15-3.07 (m, 6H ).

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HPLC-MS (Method D): [m/z]: 425.1 [M+H] ⁺ .

2-(2-{[2-(1H-1,3-Benzodiazol-2-yl)ethyl]amino}ethyl)-N-[(3-fluoropyridin-2-yl)methyl]-1,3-thiazol- 4carboxamide trihydrochloride (illustrative example No. 94- HC salt)

A 4M solution of HC in 1,4-dioxane (2.38 ml, 9.82 mmol) was added to a solution of 2-(2-{[2-(1H-1,3benzodiazol-2-yl)ethyl]amino}ethyl)- N-[(3-fluoropyridin-2-yl)methyl]-1,3-thiazole-4-carboxamide (Illustrative Example No. 94) (1.30 g, 2.98 mmol) in MeOH (15 ml), and reaction the mixture was stirred at room temperature for 2 hours. The resulting mixture was evaporated in vacuo to give the title compound (1.16 g, 70%) as an off-white solid.

1H NMR (Methanol-d4, 500 MHz): δ [ppm] = 8.48 (s, 1H), 8.25 (s, 1H), 8.07 (s, 1H), 7.79 ( dt, J=6.9, 3.4 Hz, 2H), 7.73 (s, 1H), 7.61 (dd, J=6.2, 3.1 Hz, 2H), 4.91 (s , 2H), 3.82 (s, 4H), 3.75 (t, J=6.4 Hz, 2H), 3.61 (t, J=6.4 Hz, 2H).

HPLC-MS (Method D): [m/z]: 425.1 [M+H] ⁺ .

tert-Butyl

2-[({2-[4-(benzylcarbamoyl)-1,3-thiazol-2-yl]ethyl}amino)methyl]-5-methyl-1H-1,3 benzodiazole-1-carboxylate( 116)

DIPEA (0.138 ml, 1 mmol) was added to a solution of 2-(2-aminoethyl)-N-benzyl-1,3-thiazole-4-carboxamide (104) (186 mg, 0.71 mmol) in DMF (5 ml) , then added tert-butyl 2-(chloromethyl)-5-methyl-1H-1,3benzodiazol-1-carboxylate (F) (200 mg, 0.71 mmol) and the reaction mixture was heated at 90°C. After completion (LCMS), the resulting mixture was evaporated in vacuo. The residue was purified by flash column chromatography (eluting with DCM/MeOH 95:5) as a yellow oil (85 mg, 24%).

1H NMR (CDCl ₃ , 400 MHz): δ [ppm] = 7.95 (s, 1H), 7.61 (s, 1H), 7.31 (dd, J=24.4, 4, 2 Hz, 6H), 7.07 (d, J=8.2 Hz, 1H), 4.61 (d, J=6.1 Hz, 2H), 4.58 (s, 2H), 3.74 (t, J=6.6 Hz, 2H), 3.20 (t, J=6.6 Hz, 2H), 2.46 (s, 3H), 1.36 (s, 9H).

HPLC-MS (Method I): [m/z]: 506.6 [M+H] ⁺ .

tert-Butyl 2-[({2-[4-(benzylcarbamoyl)-1,3-thiazol-2-yl]ethyl}amino)methyl]-5-methoxy-1H-1,3benzodiazol-1-carboxylate( 117)

N-benzyl-2-(2-{[(5-methoxy-1H-1,3-benzodiazol-2i yl)methyl]amino}ethyl)-1,3-thiazole-4-carboxamide (118)

Similar to general procedure 7, 2-(2-aminoethyl)-N-benzyl-1,3-thiazole-4-carboxamide (104) (0.4 g, 2 mmol), tert-butyl 2-(chloromethyl)-5-methoxy -1H-1,3-benzodiazol-1-carboxylate (F) (0.45 g, 2 mmol), DIPEA (0.4 g, 3 mmol) and NaI (0.23 g, 2 mmol) in MeCN (30 ml) under argon atmosphere at 90° C. for 18 h gave a mixture of a red solid (boc-deleted product, 350 mg, 27%) and the expected product as a white solid (133 mg, 21%) after purification by method flash column chromatography (eluting with a DCM/MeOH 5-7% gradient).

HPLC-MS (Method I): [m/z]: 522.6 [M+H]+ and 422.6 [M+H] ⁺ .

2-{2-[(1,3-Benzoxazol-2-ylmethyl)amino]ethyl}-N-[(3-fluoropyridin-2-yl)methyl]-1,3-thiazole-4carboxamide (Illustrative Example No. 115)

Similar to general procedure 7, 2-(2-aminoethyl)-^[(3-fluoropyridin-2-yl)methyl]-1,3-thiazole-4-carboxamide dihydrochloride (103) (300 mg, 0.735 mmol), 2-(chloromethyl )-1,3-benzoxazole (160 mg, 0.956 mmol), DIPEA (1.922 ml, 11.03 mmol) and DMF (15 ml) at 30°C for 24 h gave the title compound (102 mg, 34% ) as a yellow oil after purification by basic prep-HPLC.

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1H NMR (Methanol-d ₄ , 500 MHz): δ [ppm] = 8.30 (d, J=4.7 Hz, 1H), 8.08 (s, 1H), 7.66-7 .62 (m, 1H), 7.607.54 (m, 2H), 7.39-7.32 (m, 3H), 4.77 (d, J=1.6 Hz, 2H), 4.14 ( s, 2H), 3.27 (d, J=6.3 Hz, 2H), 3.18 (t, J=6.6 Hz,

2H).

HPLC-MS (Method D): [m/z]: 412.1 [M+H]+.

2-(2-{[2-(1H-1,3-Benzodiazol-2-yl)ethyl]amino}ethyl)-N-(pyridin-2-ylmethyl)-1,3-thiazole-4carboxamide (illustrative example no. 58) about

^Ν __ ην-ΝΝ vy

Similar to general procedure 7, 2-(2-aminoethyl)-N-(pyridin-2-ylmethyl)-1,3-thiazol-4-carboxamide dihydrochloride (105) (240 mg, 0.72 mmol), 2-(2 -chloroethyl)-1H-1,3-benzodiazol (259 mg, 1.43 mmol) and DIPEA (2.17 ml, 12.53 mmol) in DMF (10 ml) gave the title compound (64 mg, 22% ) as a brown solid after purification by basic prep-HPLC followed by flash column chromatography (gradient elution with 0-10% MeOH/DCM followed by 0.8 M ammonia in MeOH/DCM).

1H NMR (DMSO-d ₆ , 500 MHz): δ [ppm] = 8.87 (t, J=6.0 Hz, 1H), 8.50 (d, J=4.6 Hz, 1H ), 8.10 (s, 1H), 7.74 (td, J=7.7, 1.7 Hz, 1H), 7.44 (s, 2H), 7.33-7.21 (m, 2H), 7.10 (dd, J=5.9, 3.2 Hz, 2H), 4.55 (d, J=6.0 Hz, 2H), 3.15 (t, J=6.7 Hz, 2H), 3.02 (t, J=6.7 Hz, 2H), 2.90-2.96 (m, 4H).

HPLC-MS (method d): [m/z]: 407.2 [M+H] ⁺ .

2-(2-{[2-(1H-1,3-Benzodiazol-2-yl)ethyl](methyl)amino}ethyl)-N-[(3-fluoropyridin-2-yl)methyl]-1,3thiazole -4-carboxamide trihydrochloride (illustrative example No. 112)

2-(2-{[2-(1H-1,3-benzodiazol-2-yl)ethyl]amino}ethyl)-N-[(3-fluoropyridin-2-yl)methyl]-1,3-thiazol- 4-carboxamide trihydrochloride (illustrative example No. 94) (114 mg, 0.205 mmol), Et ₃ N (0.143 ml, 1.025 mmol) and DMF (1 ml) were stirred at room temperature for 1 h. MeI (0.059 ml, 0.949 mmol) was added and stirred at room temperature for 140 hours. Water (10 ml) was added and the solvent was evaporated in vacuo. The crude product was purified by basic prep-HPLC to give the free base (18 mg). MeOH (2 mL) and 4M HC in dioxane (0.05 mL, 0.205 mmol) were added and stirred at room temperature for 2 h. The reaction was evaporated in vacuo to give the title compound (24 mg, 21%) in form of a yellow solid.

1H NMR (Methanol-d ₄ , 500 MHz): δ [ppm] = 8.48-8.39 (bdm, 1H), 8.26 (s, 1H), 8.02-7, 84 (bm, 1H), 7.82-7.76 (m, 2H), 7.67-7.54 (m, 3H), 4.84 (s, 2H, overlap with water peak), 3 .99-3.84 (m, 6H), 3.71 (t, J=6.8 Hz, 2H), 3.16 (s, 3H).

HPLC-MS (Method D): [m/z]: 439.1 [M+H] ⁺ .

2-{2-[2-(1H-1,3-Benzodiazol-2-yl)acetamido]ethyl}-N-[(3-fluoropyridin-2-yl)methyl]-1,3-thiazole-4carboxamide dihydrochloride ( illustrative example No. 113)

Similar to general procedure 6, 2-(2-aminoethyl)-N-[(3-fluoropyridin-2-yl)methyl]-1,3-thiazole-4-carboxamide dihydrochloride (103) (150 mg, 0.368 mmol), 2-( 1H-1,3-benzodiazol-2-yl)acetic acid (114 mg, 0.552 mmol), DIPEA (0.384 ml, 2.206 mmol) and HATU (210 mg, 0.52 mmol) in THF (20 ml) at room temperature within 2 hours gave the title compound as the free base (73 mg) after purification by basic prep-HPLC. The resulting free base and a 12M solution of HC (2 ml) in MeOH (6 ml) were stirred at room temperature for 2 h. The reaction mixture was evaporated in vacuo to give the title compound (97 mg, 51%) as a brown solid.

1 H NMR (Methanol-d ₄ , 500 MHz): δ [ppm] = 8.60 (dd, J=5.5, 1.2 Hz, 1H), 8.32 (td, J=8, 9, 1.1 Hz, 1H), 8.15 (s, 1H), 7.96-7.90 (m, 1H), 7.81-7.75 (m, 2H), 7.63-7 .58 (m, 2H), 4.93 (d, J=1.1 Hz, 2H), 4.27 (s, 2H), 3.76 (t, J=6.7 Hz, 2H), 3 .32 (t, J=6.6 Hz, 2H).

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HPLC-MS (Method D): [m/z]: 439.1 [M+H]+.

2-{2-[(Cyclopropylmethyl)amino]ethyl}-N-[(3-fluoropyridin-2-yl)methyl]-1,3-thiazole-4 (119) carboxamide

Similar to general procedure 3, 2-(2-aminoethyl)-N-[(3-fluoropyridin-2-yl)methyl]-1,3-thiazole-4-carboxamide dihydrochloride (103) (56% pure, 500 mg, 0.793 mmol) , cyclopropanecarbaldehyde (67 mg, 0.951 mmol) and DIPEA (0.552 ml, 3.17 mmol) in MeOH (7 ml) at room temperature for 16 h after addition of NaBH4 (45 mg, 1.19 mmol) gave the indicated in title compound (149 mg, 51%) as a colorless oil after purification by flash column chromatography (kp-NH, eluting with a 0-10% MeOH/DCM gradient).

¹ H NMR (DMSO-d6, 500 MHz): δ [ppm] = 8.65 (t, J=5.7 Hz, 1H), 8.39 (dt, J=4.5, 1, 3 Hz, 1H), 8.12 (s, 1H), 7.71 (ddd, J=10.0, 8.3, 1.3 Hz, 1H), 7.41 (dt, J=8.6 , 4.5 Hz, 1H), 4.66 (dd, J=5.7, 1.4 Hz, 2H), 3.13 (t, J=6.7 Hz, 2H), 2.92 (t , J=6.7 Hz, 2H), 2.43 (d, J=6.6 Hz, 2H), 1.92 (s, 1H), 0.93-0.81 (m, 1H), 0 .43-0.35 (m, 2H),

0.13-0.06 (m, 2H).

HPLC-MS (method F): [m/z]: 335.8 [M+H] ⁺ .

2-(2-{[2-(1H-1,3-Benzodiazol-2-yl)ethyl](cyclopropylmethyl)amino}ethyl)-X-[(3-fluoropyridin-2yl)methyl]-1,3-thiazole -4-carboxamide (illustrative example No. 148)

Similar to general procedure 7, 2-{2-[(cyclopropylmethyl)amino]ethyl}-H-[(3-fluoropyridin-2yl)methyl]-1,3-thiazole-4-carboxamide (119) (149 mg, 0. 45 mmol), 2-(2-chloroethyl)-1H-1,3-benzodiazol hydrochloride (116 mg, 0.53 mmol) and DIPEA (0.4 ml, 2.23 mmol) at 30°C for 32 h gave the title compound (5 mg, 2%) as a yellow oil after purification by basic prep-HPLC and then by flash column chromatography (gradient elution 0-10% MeOH in DCM followed by 0-10% 7 M ammonia solution in MeOH/DCM).

1H NMR (Acetone-d6, 500 MHz): δ [ppm] = 8.48 (s, 1H), 8.39 (d, J=4.7 Hz, 1H), 7.98 (s, 1H), 7.66-7.54 (m, 1H), 7.49-7.33 (m, 3H), 7.10 (dd, J=6.0, 3.2 Hz, 2H), 4 .75 (dd, J=5.3, 1.5 Hz, 2H), 3.33 (t, J=6.7 Hz, 2H), 3.30

3.11 (m, 6H), 2.67 (d, J=6.7 Hz, 2H), 1.10-0.94 (m, 1H), 0.60-0.43 (m, 2H) , 0.26-0.22 (m, 2H).

HPLC-MS (method B): [m/z]: 479.2 [M+H] ⁺ .

2-(2-{[2-(1H-1,3-Benzodiazol-2-yl)ethyl]amino}ethyl)-X-(pyridazin-3-ylmethyl)-1,3-thiazole-4carboxamide trihydrochloride (illustrative example No. 122)

Similarly, using the general procedure 7, 2-(2-aminoethyl)-X-(pyridazin-3-ylmethyl)1,3-thiazol-4-carboxamide dihydrochloride (106) (409 mg, 1.22 mmol), 2- (2-chloroethyl)-1H-1,3-benzodiazol hydrochloride (316.9 mg, 1.46 mmol) and DIPEA (3.18 ml, 0.02 mol) in DMF (5 ml) at room temperature for 5 days gave the free base product after purification by flash column chromatography eluting with a 0-20% MeOH/DCM gradient, followed by purification by basic prep-HPLC.

The resulting free base product was redissolved in MeOH (5 mL) and 12M HC (1 mL) was added over 1 hour to give the title compound (132 mg, 21%) as a pale yellow solid.

1H NMR (Methanol-d4, 500 MHz): δ [ppm] = 9.52 (dd, J=5.2, 1.2 Hz, 1H), 8.59 (dd, J=8.7 , 1.2 Hz, 1H), 8.46 (dd, J=8.7, 5.2 Hz, 1H), 8.28 (s, 1H), 7.83 (dd, J=6.2, 3.1 Hz, 2H), 7.65 (td, J=6.2, 5.5, 2.2 Hz, 2H), 5.04 (s, 2H), 3.90-3.82 (m , J=4.2 Hz, 4H), 3.79 (t, J=6.4 Hz, 2H), 3.64 (t, J=6.4 Hz, 2H).

HPLC-MS (Method C): [m/z]: 408.2 [M+H] ⁺ .

2-(2-{[2-(1H-1,3-Benzodiazol-2-yl)ethyl]amino}ethyl)-X-(pyrimidin-2-yl-methyl)-1,3-thiazole-4carboxamide trihydrochloride ( illustrative example No. 129)

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Similar to general procedure 7, 2-(2-aminoethyl)-N-(pyrimidin-2-ylmethyl)-1,3-thiazol-4-carboxamide dihydrochloride (108) (300 mg, 0.89 mmol), 2-(2-chloroethyl )-1H-1,3-benzodiazol hydrochloride (193.7 mg, 0.89 mmol) and DIPEA (3.11 ml, 17.8 mmol) in DMF (10 ml) at room temperature for 9 days gave the product in free base form after purification by flash column chromatography (elution with 0-40% MeOH/DCM gradient), followed by purification by basic prep-HPLC.

The resulting free base product was redissolved in MeOH (5 ml) and 12M HC was added over 30 minutes to give the title compound (26 mg, 6%) as a pale yellow solid.

¹ H NMR (D ₂ O, 500 MHz): δ [ppm] = 8.66 (d, J=5.1 Hz, 2H), 8.15 (s, 1H), 7.67 (dt , J=6.7, 3.4 Hz, 2H), 7.53 (td, J=6.2, 5.5, 2.1 Hz, 2H), 7.40 (t, J=5.1 Hz, 1H), 4.60 (s, 2H), 3.76-3.65 (m, 6H), 3.54 (t, J=6.4 Hz, 2H).

HPLC-MS (Method E): [m/z]: 408.1 [M+H]+.

General Procedure 8: 2-(2-{[2-(1H-1,3-Benzodiazol-2-yl)ethyl]amino}ethyl)-N-[(5-methylpyrimidin-2yl)methyl]-1,3- thiazole-4-carboxamide (illustrative example No. 155)

DBU (15.37 µl, 0.1 mmol) was added to a suspension of 2-(2-aminoethyl)-X-[(5-methylpyrimidin-2yl)methyl]-1,3-thiazole-4-carboxamide dihydrochloride (109) ( 36 mg, 0.1 mmol) in MeCN (3 ml). N-(2-nitrophenyl)prop-2-enamide (D) (19 mg, 0.1 mmol) was added and the reaction mixture was stirred at room temperature overnight. The reaction mixture was diluted with EtOAc (5 ml) and washed with 10% NaHCO ₃ solution (5 ml), water (5 ml), saturated sodium chloride solution (5 ml), dried (MgSO ₄ ), filtered and evaporated to give crude an intermediate which was further reacted with iron powder (3 mg, 0.05 mmol) in AcOH (3 ml) at 80°C for 3 h. The reaction mixture was diluted with water (5 ml), then basified by slow addition of 10 M aqueous NaOH . The resulting mixture was then further diluted with water (10 ml) and extracted with a 4:1 mixture of chloroform/IPS (4x30 ml). The combined organic layers were separated, dried (MgSO ₄ ) and evaporated in vacuo. The resulting crude product was purified by basic prep-HPLC to give the title compound (8 mg, 67%) as a colorless film.

1H NMR (Acetone-de, 500 MHz): δ [ppm] = 8.58 (s, 2H), 8.44 (s, 1H), 8.03 (s, 1H), 7.45 ( s, 2H), 7.14-7.05 (m, 2H), 4.72 (d, J=4.3 Hz, 2H), 3.24 (t, J=6.5 Hz, 2H), 3.13 (m, 4H), 3.07 (t, J=6.2 Hz, 2H), 2.29 (s, 3H).

HPLC-MS (Method C): [m/z]: 422.0 [M+H] ⁺ .

N-{5Н,6Н,7Н-cyclopenta[b]pyridin-7-yl}-2-[2-({2-[(2-nitrophenyl)karbamoyl]ethyl}amino)ethyl]1,3-thiazol-4 -carboxamide (illustrative example No. 158)

Similar to general procedure 8, 2-(2-aminoethyl)-N-{5H,6H,7H-cyclopenta[b]pyridin-7-yl}-1,3thiazol-4-carboxamide dihydrochloride (110) (660 mg, 1, 83 mmol), N-(2-nitrophenyl)prop-2-enamide (D) (344 mg, 1.79 mmol) and DBU (0.8 ml, 5.37 mmol) in MeCN (8 ml) gave the crude intermediate product which reacted with iron powder (180 mg, 3.22 mmol) in AcOH (10 ml) to give the title compound (176 mg, 25%) as a pale yellow foam after purification by flash column chromatography (gradient elution 5-10% 3M ammonia in MeOH/DCM), followed by a basic prep-HPLC.

1H NMR (CDCl ₃ , 500 MHz): δ [ppm] = 8.27 (d, J=4.7 Hz, 1H), 8.17 (s, 1H), 8.07 (br.s , 1H), 7.43 (brs, 2H), 7.37-7.32 (m, 1H), 7.19 (dt, J=8.5, 4.4 Hz, 1H), 7, 15-7.11 (m, 2H), 4.73 (dd, J=5.0, 1.3 Hz, 2H), 3.18 (t, J=6.2 Hz, 2H), 3.17 -3.08 (m, 4H), 3.04 (t, J=6.2 Hz, 2H).

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HPLC-MS (method C): [m/z]: 433.2 [M+H]+.

2-(2-{[2-(1H-1,3-Benzodiazol-2-yl)ethyl]amino}ethyl)-N-[(6-methylpyridazin-3-yl)methyl]-1,3-thiazol4- carboxamide (illustrative example No. 174)

Similar to general procedure 8, 2-(2-Aminoethyl)-N-[(6-methylpyridazin-3-yl)methyl]-1,3-thiazole-4-carboxamide dihydrochloride (107) (382 mg, 0.932 mmol), N-( 2-nitrophenyl)prop-2-enamide (D) (161 mg, 0.839 mmol) and DBU (0.300 ml, 2.01 mmol) in MeCN (15 ml) at room temperature over 2 h gave the desired Michael intermediate (163 mg , 31%) as a yellow oil after purification by flash column chromatography (0-3% MeOH/DCM), followed by a second purification on an Isolute silica gel column with a gradient of 0-2% 7M NH ₃ /MeOH in DCM.

The Michael intermediate (163 mg, 0.288 mmol) was reacted with iron powder (32 mg) in AcOH (3 mL) at 80°C for 1 h to give the title compound (15 mg, 12%) as a beige solid after purification by basic prep-HPLC, followed by column chromatography on kp-NH silica gel.

¹ H NMR (DMSO-d6, 500 MHz): δ [ppm] = 9.00 (t, J=6.0 Hz, 1H), 8.10 (s, 1H), 7.52-7 .42 (m, 4H), 7.137.09 (m, 2H), 4.70 (d, J=6.1 Hz, 2H), 3.15 (t, J=6.7 Hz, 2H), 3 .04-3.00 (m, 2H), 3.00-2.94 (m, 4H), 2.59 (s, 3H).

HPLC-MS (method B): [m/z]: 422.2 [M+H] ⁺ .

2-(2-{[2-(1H-1,3-Benzodiazol-2-yl)ethyl]amino}ethyl)-N-[(3-fluoropyridin-2-yl)methyl]-5-methyl-1, 3thiazole-4-carboxamide (illustrative example No. 165)

Similar to general procedure 8, 2-(2-aminoethyl)-X-[(3-fluoropyridin-2-yl)methyl]-5-methyl-1,3thiazole-4-carboxamide dihydrochloride (113) (528 mg, 1.44 mmol), N-(2-nitrophenyl)prop-2-enamide (D) (276 mg, 1.44 mmol) and DBU (0.64 ml, 0 mol) in MeCN (20 ml) at room temperature for 16 gave a crude Michael intermediate (50%, 697 mg, 0.72 mmol) which was further reacted with iron powder (40 mg) in AcOH (4 mL) at 80°C for 1.5 h to give the title compound (99 mg, 32%) as a pale yellow solid after purification by flash column chromatography eluting with 2-40% MeOH in DCM, followed by prep-HPLC.

¹ H NMR (DMSO-d ₆ , 500 MHz): δ [ppm] = 8.57 (t, J=5.6 Hz, 1H), 8.38 (dt, J=4.3, 1 .3 Hz, 1H), 7.73-7.66 (m, 1H), 7.45 (dd, J=5.7, 3.2 Hz, 2H), 7.40 (dt, J=8, 6, 4.4 Hz, 1H), 7.13-7.08 (m, 2H), 4.61 (d, J=5.5 Hz, 2H), 3.06 (q, J=6.3 Hz, 4H), 2.97 (q, J=6.7 Hz, 4H), 2.60 (s, 3H).

HPLC-MS (method C): [m/z]: 439.2 [M+H] ⁺ .

2-(2-{[2-(1H-1,3-Benzodiazol-2-yl)ethyl]amino}ethyl)-5-methyl-X-(pyrimidin-2-ylmethyl)-1,3-thiazole4-carboxamide (illustrative example #188)

Similar to general procedure 8, 2-(2-aminoethyl)-5-methyl-N-(pyrimidin-2-ylmethyl)-1,3-thiazol-4-carboxamide dihydrochloride (114) (315 mg, 0.594 mmol), X-(2 -nitrophenyl)prop-2-enamide (D) (114 mg, 0.594 mmol) and DBU (0.266 ml, 1.781 mmol) in MeCN (12 ml) at room temperature for 3 hours gave the target Michael intermediate (142 mg, 42% ) as a yellow oil after purification using an Isolute silica gel column eluting with a gradient of 0-6% MeOH in DCM.

The Michael intermediate (142 mg, 0.248 mmol) was reacted with iron powder (42 mg) in AcOH (3 mL) at 80°C for 1.5 h to give the title compound (7 mg, 7%) as a brown solid substances after purification by basic prep-HPLC, followed by purification on an Isolute silica gel column eluting with a gradient of 0-8% MeOH in DCM.

1H NMR (Methanol-d ₄ , 500 MHz): δ [ppm] = 8.72 (d, J=4.9 Hz, 2H), 7.43 (dt, J=6.6, 3, 3 Hz, 2H), 7.35 (t,

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J=4.9Hz, 1H), 7.19(dt, J=6.0, 3.4Hz, 2H), 4.65(s, 2H), 3.24(t, J=6.8 Hz, 2H), 3.22-3.17 (m, 4H), 3.15 (t,

J=6.6 Hz, 2H), 2.68 (s, 3H).

HPLC-MS (Method D): [m/z]: 422.2 [M+H]+.

Benzyl N-(3-{[2-( 1 H-1,3-benzodiazol-2-yl)ethyl] [2-(4- {[(3-fluoropyridin-2-yl)methyl]carbamoyl} -1, 3thiazol-2-yl)ethyl]amino}propyl)carbamate (illustrative example No. 179)

Similar to general procedure 3, 2-(2-{[2-(1H-1,3-benzodiazol-2-yl)ethyl]amino}ethyl)-N-[(3fluoropyridin-2-yl)methyl]-1,3 -thiazole-4-carboxamide (illustrative example No. 94 - free base) (166 mg, 0.391 mmol), benzyl (3-oxopropyl) carbamate (97 mg, 0.469 mmol) and DIPEA (0.12 ml, 0.587 mmol) in MeOH (1 ml) at room temperature for 1 h, after adding NaBH ₄ (22 mg, 0.587 mmol), gave the title compound (94 mg, 39%) as a pale yellow oil after purification by flash column chromatography ( eluting with a gradient of 0-5% MeOH/DCM).

HPLC-MS (method F): [m/z]: 616.2 [M+H] ⁺ .

2-{2-[(3-Aminopropyl)[2-(1H-1,3-benzodiazol-2-yl)ethyl]amino]ethyl}-N-[(3-fluoropyridin-2yl)methyl]-1,3 -thiazole-4-carboxamide (illustrative example No. 163)

Benzyl solution N-(3-{[2-(1H-1,3-benzodiazol-2-yl)ethyl][2-(4-{[(3-fluoropyridin-2yl)methyl]carbamoyl}-1,3- thiazol-2-yl)ethyl]amino}propyl)carbamate (illustrative example No. 179) (45 mg, 0.073 mmol) in AcOH/HBr (1:1, 1 ml) was stirred at 50°C for 2 hours. The reaction mixture evaporated in vacuum. Purification by basic prep-HPLC gave the title compound (16 mg, 45%) as a colorless oil.

¹ H NMR (Methanol ₄ , 500 MHz): δ [ppm] = 8.32 (d, J=4.7 Hz, 1H), 7.90 (s, 1H), 7.63- 7.52 (m, 1H), 7.44 (dt, J=6.6, 3.3 Hz, 2H), 7.35 (dt, J=8.6, 4.4 Hz, 1H), 7 .17 (dt, J=6.0, 3.3 Hz, 2H), 4.75 (d, J=1.5 Hz, 2H), 3.20 (t, J=6.6 Hz, 2H) , 3.04 (s, 4H), 2.98 (t, J=6.6 Hz, 2H), 2.65 (m, 4H), 1.67 (m, 2H).

HPLC-MS (Method D): [m/z]: 482.2 [M+H] ⁺ .

2-(2-{[2-(1H-1,3-Benzodiazol-2-yl)ethyl](butyl)amino}ethyl)-N-[(3-fluoropyridin-2-yl)methyl]-1,3thiazole -4-carboxamide (illustrative example No. 157)

Similar to general procedure 3, 2-(2-{[2-(1H-1,3-benzodiazol-2-yl)ethyl]amino}ethyl)-N-[(3fluoropyridin-2-yl)methyl]-1,3 -thiazole-4-carboxamide (illustrative example No. 94 - free base) (60 mg, 0.141 mmol), butanal (12 mg, 0.174 mmol) and DIPEA (98 μl, 0.56 mmol) in MeOH (1 ml) at room temperature for 1 h, after adding NaBH ₄ (8 mg, 0.21 mmol), gave the title compound (50 mg, 73%) as a yellow oil after purification by basic prep-HPLC.

1H NMR (DMSO-a ₆ , 500 MHz): δ [ppm] = 12.13 (s, 1H), 8.66 (t, J=5.6 Hz, 1H), 8.37 (d , J=4.7 Hz, 1H), 8.06 (s, 1H), 7.69 (ddd, J=9.9, 8.4, 1.2 Hz, 1H), 7.57-7, 32 (m, 3H), 7.17-7.01 (m, 2H), 4.65 (d, J=4.6 Hz, 2H), 3.17 (t, J=6.7 Hz, 2H ), 2.96 (m, 4H), 2.88 (t, J=6.7 Hz, 2H), 2.58-2.51 (m, 2H), 1.40 (m, 2H), 1 .23 (m, 2H), 0.83 (t, J=7.4 Hz, 3H).

HPLC-MS (Method D): [m/z]: 481.3 [M+H] ⁺ .

2-(2-{bis[2-(1H-1,3-Benzodiazol-2-yl)ethyl]amino}ethyl)-N-[(3-fluoropyridin-2-yl)methyl]-1,3thiazol-4 -carboxamide (illustrative example No. 152)

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Similar to general procedure 7, 2-(2-{[2-(1H-1,3-benzodiazol-2-yl)ethyl]amino}ethyl)-N-[(3fluoropyridin-2-yl)methyl]-1,3 -thiazole-4-carboxamide (illustrative example No. 94 - free base) (400 mg, 1.13 mmol), 2-(2-chloroethyl)-1H-1,3-benzodiazol hydrochloride (492 mg, 2.27 mmol) and DIPEA (3.03 ml, 17 mol) in DMF (5 ml) at 30°C for 3 days gave the title compound (10 mg, 1.5%) as an off-white solid after purification by basic prep-HPLC, followed by flash column chromatography (gradient elution 0-10% MeOH/DCM, then 010% 7N ammonia in MeOH/DCM) and a second main prep-HPLC.

1H NMR (DMSO-66, 500 MHz): δ [ppm] = 12.28 (s, 1H), 8.66 (t, J=5.7 Hz, 1H), 8.37 (dt, J=4.6, 1.4Hz, 1H), 8.06(s, 1H), 7.69(ddd, J=10.0, 8.3, 1.3Hz, 1H), 7.61 -7.46 (m, 3H), 7.39 (d.s., J=8.6, 4.2 Hz, 2H), 7.237.04 (m, 4H), 4.77-4.52 ( m, 4H), 3.27 (t, J=7.2 Hz, 2H), 3.15-3.04 (m, 2H), 3.02-2.90 (m, 4H), 2.87 (t, J=6.7 Hz, 2H).

HPLC-MS (method B): [m/z]: 569.3 [M+H] ⁺ .

2-{2-[bis(1H-1,3-Benzodiazol-2-ylmethyl)amino]ethyl}-N-[(3-fluoropyridin-2-yl)methyl]-1,3-thiazole4-carboxamide (illustrative example) No. 166)

Similar to general procedure 7, 2-{2-[(1H-1,3-benzodiazol-2-ylmethyl)amino]ethyl}-N-[(3fluoropyridin-2-yl)methyl]-1,3-thiazol-4- carboxamide (illustrative example No. 40) (110 mg, 0.268 mmol), 2-(chloromethyl)-1H-1,3-benzodiazol (45 mg, 0.268 mmol), DIPEA (0.467 ml, 2.68 mmol) in DMF (1 ml) at 45°C for 3 h, and then at 55°C for 1 h, gave the title compound (10 mg, 7%) as a white solid after purification by basic prep-HPLC, followed by column flash chromatography (gradient elution with 0-10% MeOH/DCM followed by 0-10% 7N ammonia in MeOH/DCM).

¹ H NMR (Acetone-d6, 500 MHz): δ [ppm] = 11.99 (s, 1H), 8.43 (s, 1H), 8.33 (d, J=4.7 Hz , 1H), 8.02 (s, 1H), 7.71-7.55 (m, 3H), 7.51 (s, 2H), 7.37 (dt, J=8.5, 4.4 Hz, 1H), 7.17 (d, J=5.7 Hz, 4H), 4.80-4.70 (m, 2H), 4.17 (s, 4H), 3.37 (t, J =6.9 Hz, 2H), 3.20 (t, J=6.9 Hz, 2H).

HPLC-MS (method B): [m/z]: 541.3 [M+H] ⁺ .

2-(2-{[2-(4-Fluoro-1H-1,3-benzodiazol-2-yl)ethyl]amino}ethyl)-N-[(3-fluoropyridin-2-yl)methyl]-1, 3thiazole-4-carboxamide (illustrative example No. 243)

Similar to general procedure 8, 2-(2-Aminoethyl)-N-[(3-fluoropyridin-2-yl)methyl]-1,3-thiazole-4-carboxamide dihydrochloride (103) (662 mg, 1.87 mmol), N -(3-fluoro-2-nitrophenyl)prop-2-enamide (G) (394 mg, 1.87 mmol) and DBU (924 μl, 6.18 mmol) in MeCN (10 ml) gave a crude intermediate which reacted with iron powder (286 mg, 5.12 mmol) in AcOH (15 mL) to give the title compound (203 mg, 34%) as a white solid after purification by basic pre-HPLC, followed by flash column chromatography ( eluting with a gradient of 0-20% MeOH/DCM).

Ή NMR (DMSO-d ₆ , 500 MHz): δ [ppm] = 8.66 (t, J=5.6 Hz, 1H), 8.39 (dt, J=4.7, 1, 3 Hz, 1H), 8.08 (s, 1H), 7.70 (ddd, J=10.0, 8.4, 1.1 Hz, 1H), 7.40 (dt, J=8.7 , 4.4 Hz, 1H), 7.27 (d, J=8.0 Hz, 1H), 7.10 (td, J=8.0, 4.9 Hz, 1H), 6.92 (dd , J=11.1, 8.0 Hz, 1H), 4.68-4.62 (m, 2H), 3.16 (t, J=6.7 Hz, 2H), 3.05 (t, J=6.7 Hz, 2H), 2.99 (t, J=6.6 Hz, 4H).

HPLC-MS (method B): [m/z]: 443.2 [M+H]+.

2-(2-{[(4-fluoro-1H-1,3-benzodiazol-2-yl)methyl]amino}ethyl)-N-[(3-fluoropyridin-2-yl)methyl]-1,3oxazol- 4-carboxamide (illustrative example No. 253)

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Similar to general procedure 7, 2-(2-aminoethyl)-N-[(3-fluoropyridin-2-yl)methyl]-1,3-oxazole-4carboxamide dihydrochloride (235) (100 mg, 0.3 mmol), 2 -(chloromethyl)-7-fluoro-1H-1,3-benzodiazol hydrochloride (66 mg, 0.3 mmol) and DIPEA (258 μl, 1.48 mmol) in DMF (2.5 ml) were stirred at 40°C over 73 hours to give the title compound (23 mg, 18%) as a brown glassy solid after purification on a reverse phase Biotage column (A=water/0.1% NH3; B=MeCN/0.1% NH3; gradient elution of 10% A/B for 2 column volumes, 10-30% A/B for 4 column volumes, 30-60% A/B for 10 column volumes, and 60-100% for 5 column volumes).

Ή NMR (DMSO-d6, 500 MHz): δ [ppm] = 12.49 (brs, 1H), 8.54-8.47 (m, 2H), 8.39-8.34 (m, 1H), 7.737.65 (m, 1H), 7.42-7.37 (m, 1H), 7.34-7.24 (m, 1H), 7.15-7.06 (m , 1H), 6.92 (t, 1H), 4.63-4.57 (m, 2H), 3.99-3.91 (m, 2H), 3.02-2.94 (m, 4H ), 2.63 (broad s, 1H).

HPLC-MS (Method D): [m/z]: 413.2 [M+H]+.

2-(2-{[(4-fluoro-1H-1,3-benzodiazol-2-yl)methyl]amino}ethyl)-N-[(3-fluoropyridin-2-yl)methyl]-1,3thiazol- 4-carboxamide (illustrative example No. 274)

Similar to general procedure 7, 2-(2-Aminoethyl)-N-[(3-fluoropyridin-2-yl)methyl]-1,3-thiazole-4-carboxamide dihydrochloride (103) (200 mg, 0.57 mmol), 2 -(chloromethyl)-7-fluoro-1H-1,3-benzodiazol hydrochloride (125 mg, 0.57 mmol) and DIPEA (493 μl, 2.83 mmol) in DMF (5.5 ml) were heated at 40°C for 18 h, stirred at room temperature for 2 days and then heated at 40° C. for 4 h to give the title compound (7 mg, 3%) as a brown glassy solid after purification by reverse phase chromatography [(elution gradient 10-100% (water+0.1% ammonia)/(MeCM+0.1% ammonia)].

1H NMR (DMSO-H6, 500 MHz): δ [ppm] = 12.50 (s, 1H), 8.70-8.61 (m, 1H), 8.39-8.32 (m , 1H), 8.13 (s, 1H), 7.72-7.66 (m, 1H), 7.42-7.36 (m, 1H), 7.34-7.25 (m, 1H ), 7.15-7.08 (m, 1H), 6.93 (m, 1H), 4.68-4.62 (m, 2H), 4.00-3.94 (m, 2H), 3.22-3.15 (m, 2H), 3.01-2.93 (m, 2H), 2.71 (s, 1H).

HPLC-MS (Method D): [m/z]: 429.1 [M+H] ⁺ .

The above general scheme 6:

general technique 9:

2-{2-[(1H-1,3-Benzodiazol-2-yl)amino]ethyl}-H-[(3-fluoropyridin-2-yl)methyl]-1,3-thiazole-4carboxamide dihydrochloride (illustrative example) No. 121)

2-(2-Aminoethyl)-N-[(3-fluoro-pyridin-2-yl)methyl]-1,3-thiazole-4-carboxamide dihydrochloride solution (103) (1.65 g, 2.62 mmol, 56% purity), 2-chloro-1H-benzimidazole (0.1 g, 0.66 mmol) and DIPEA (0.571 ml, 3.277 mmol) in n-butanol (3 ml) and MeOH (0.1 ml) were heated at 150° C in a microwave oven for 2.5 hours. The reaction mixture was evaporated in vacuo. The resulting residue was dissolved in saturated NaHCO ₃ solution, diluted with water (20 ml) and extracted with 4:1 chloroform/IPS (4x20 ml). The combined organic extracts were dried (MgSO ₄ ), filtered and evaporated in vacuo. The resulting residue was purified by flash column chromatography (kp-NH, 010% MeOH/EtOAc gradient elution) followed by basic prep-HPLC. The resulting residue was dissolved in MeOH (4 mL) and 12M HC (1 mL) was added over 2 hours. Evaporation in vacuo gave the title compound (0.131 g, 43%) as a white solid.

1H NMR (Methanol-d4, 500 MHz): δ [ppm] = 8.60 (d, J=5.5 Hz, 1H), 8.31 (t, J=8.3 Hz, 1H) , 8.19 (s, 1H), 7.93 (br, J=3.8 Hz, 1H), 7.38 (dt, J=7.1, 3.5 Hz, 2H), 7.28 ( dd, J=5.9, 3.2 Hz, 2H), 4.93 (s, 2H), 3.99 (t, J=6.4 Hz, 2H), 3.49 (t, J=6 .4 Hz, 2H).

HPLC-MS (Method D): [m/z]: 397.1 [M+H] ⁺ .

2-{3-[(1H-1,3-Benzodiazol-2-yl)amino]pronyl}-N-[(3-fluoropyridin-2-yl)methyl]-1,3-thiazole-4carboxamide (illustrative example no. 135)

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As in general procedure 9, 2-(3-aminopropyl)-N-[(3-fluoropyridin-2-yl)methyl]-1,3-thiazole-4-carboxamide dihydrochloride (115) (173 mg, 0.472 mmol), 2-chloro -1H-benzimidazole (60 mg, 0.393 mmol), DIPEA (0.21 ml, 1.18 mmol), n-BuOH (2 ml) and DMF (0.5 ml) at 150°C in the microwave for 1 h gave the title compound (26 mg, 16%) as an off-white solid after purification by basic prep-HPLC.

¹ H NMR (DMSO-06, 500 MHz): δ [ppm] = 10.74 (s, 1H), 8.68 (t, J=5.7 Hz, 1H), 8.37 (dt , J=4.7, 1.4 Hz, 1H), 8.15 (s, 1H), 7.70 (ddd, J=10.0, 8.3, 1.3 Hz, 1H), 7, 40 (dt, J=8.6, 4.4 Hz, 1H), 7.11 (dd, J=14.8, 7.6 Hz, 2H), 6.96-6.75 (m, 2H) , 6.67 (t, J=5.7 Hz, 1H), 4.65 (dd, J=5.7, 1.4 Hz, 2H), 3.39 (kv, J=6.7 Hz, 2H), 3.18-3.07 (m, 2H), 2.07 (m, 2H).

HPLC-MS (Method D): [m/z]: 411.2 [M+H]+.

The above general scheme 7:

ethyl 2-({[2-(1H-1,3-benzodiazol-2-yl)ethyl]amino}methyl)-1,3-thiazole-4-carboxylate (120)

Similar to general procedure 3, 2-(1H-benzimidazol-2-yl)ethanamine dihydrochloride (379 mg, 1.62 mmol), ethyl 2-formyl-1,3-thiazole-4-carboxylate (300 mg, 1.62 mmol ), DIPEA (1.13 ml, 6.48 mmol) and MgSO ₄ (100 mg) in DCM (10 ml) at room temperature for 24 h, followed by the addition of NaBH ₄ (92 mg, 2.43 mmol), gave the title compound (201 mg, 35%) as a white solid after purification by flash column chromatography (kp-NH, eluting with a 015% MeOH/EtOAc gradient).

1H NMR (DMSO-06, 500 MHz): δ [ppm] = 12.17 (s, 1H), 8.40 (s, 1H), 7.51 (d, J=7.8 Hz, 1H), 7.40 (d, J=7.1 Hz, 1H), 7.16-7.07 (m, 2H), 4.28 (q, J=7.1 Hz, 2H), 4, 05 (m, 2H), 3.04 (m, 2H), 2.99 (t, J=5.8 Hz, 2H), 2.52 (s, 2H), 1.29 (t, J=7 .1 Hz, 3H).

HPLC-MS (method A): [m/z]: 331.0 [M+H] ⁺ .

Ethyl 2-({[3-(1Н-1,3-benzodiazol-2-yl)propyl]amino}methyl)-1,3-thiazole-4-carboxylate (121)

Similar to general procedure 3, 3-(1H-benzimidazol-2-yl)propan-1-amine (568 mg, 3.24 mmol), ethyl 2-formyl-1,3-thiazole-4-carboxylate (600 mg, 3 .24 mmol), DIPEA (2.26 ml, 12.96 mmol) and MgSO ₄ (300 mg) in DCM (20 ml) at room temperature for 24 h, followed by the addition of NaBH ₄ (184 mg, 4.86 mmol) gave the title compound (570 mg, 31%, 62% pure) as a white solid after purification by flash column chromatography (kp-NH, eluting with a 010% MeOH/EtOAc gradient).

1H NMR (DMSO-a ₆ , 500 MHz): δ [ppm] = 12.14 (s, 1H), 8.39 (s, 1H), 7.50 (d, J=6.9 Hz , 1H), 7.39 (d, J=7.0 Hz, 1H), 7.14-7.05 (m, 2H), 4.28 (q, J=7.1 Hz, 2H), 3 .99 (m, 2H), 2.87 (t, J=7.6 Hz, 2H), 1.94 (q, J=7.3 Hz, 2H), 1.29 (t, J=7, 1 Hz, 3H).

HPLC-MS (method A): [m/z]: 345.00 [M+H] ⁺ .

tert-Butyl 2-(2-{[(tert-butoxy)carbonyl] ({[4-(ethoxycarbonyl)-1,3-thiazol-2yl]methyl})amino}ethyl)- 1H-1,3-benzodiazol- 1-carboxylate (122) o

H N—<

°C TH] -C o \u003d (

X about

Similar to general procedure 4, ethyl 2-({[2-(1H-1,3-benzodiazol-2-yl)ethyl]amino}methyl)-1,3thiazole-4-carboxylate (120) (201 mg, 0.608 mmol) , Boc ₂ O (146 mg, 0.669 mmol) and TEA (0.08 ml, 0.608 mmol) in THF (10 ml) at room temperature for 20 h gave the title compound

- 137 039916 (345 mg, 94%) as a colorless oil after purification by flash column chromatography (eluting with a 40-100% EtOAc/heptane gradient).

HPLC-MS (method A): [m/z]: 531.15 [M+H] ⁺ .

tert-Butyl 2-(3-{[(tert-butoxy)carbonyl] ({[4-(ethoxycarbonyl)-1,3-thiazol-2-yl]methyl} )amino} propyl)- 1H-1,3- benzodiazole-1-carboxylate (123)

Similar to general procedure 4, ethyl 2-({[3-(1H-1,3-benzodiazol-2-yl)propyl]amino}methyl)-1,3thiazole-4-carboxylate (121) (0.570 g, 1.018 mmol, 62% pure), Boc ₂ O (1.56 g, 7.124 mmol) and TEA (0.671 ml, 5.089 mmol) in THF (40 ml) were reacted at room temperature for 72 h, then Boc ₂ O (0.444 g, 2.036 mmol) for 4 h and then more Boc ₂ O (0.444 g, 2.036 mmol) for another 16 h to give the title compound (1.162 g, 48% pure, quant) as a yellow oil after purification by flash column chromatography (eluting with a 0-60% EtOAc/heptane gradient).

HPLC-MS (Method A): [m/z]: 545.15 [M+H]+.

2-({[2-(1H-1,3-Benzodiazol-2-yl)ethyl] [(tert-butoxy)carbonyl]amino}methyl)-1,3-thiazole-4carboxylic acid (124)

Similar to general procedure 5, tert-butyl 2-(2-{[(tert-butoxy)carbonyl]({[4-(ethoxycarbonyl)1,3-thiazol-2-yl]methyl})amino}ethyl)-1H- 1,3-benzodiazol-1-carboxylate (122) (385 mg, 0.73 mmol) and LiOH (87 mg, 3.63 mmol) in THF/water (25 mL/5 mL) gave the title compound ( 350 mg, 99%, 83% pure) as a white solid.

HPLC-MS (method A): [m/z]: 403.00 [M+H] ⁺ .

2-({[3-(1H-1,3-Benzodiazol-2-yl)propyl] [(tert-butoxy)carbonyl]amino}methyl)-1,3-thiazole-4carboxylic acid (125)

Similar to general procedure 5, tert-butyl 2-(3-{[(tert-butoxy)carbonyl]({[4-(ethoxycarbonyl)1,3-thiazol-2-yl]methyl})amino}propyl)-1H- 1,3-benzodiazol-1-carboxylate (123) (1.16 g, 1.02 mmol, 48% purity) and LiOH (122 mg, 5.09 mmol) in THF/water (20 ml/5 ml) gave the crude title compound (811 mg, 52% pure) as an off-white solid.

¹ H NMR (DMSO-dg, 500 MHz): δ [ppm] = 8.39 (s, 1H), 7.65 (br s, 2H), 7.37 (br s, 2H) , 4.68 (brs, 2H), 3.38 (brs, 2H), 3.00 (brs, 2H), 2.08 (brs, 2H), 1.34 (s , 9H).

HPLC-MS (method A): [m/z]: 417.05 [M+H] ⁺ .

tert-Butyl N-[2-(1H-1,3-benzodiazol-2-yl)ethyl]-N-[(4-{[(3-fluoropyridin-2-yl)methyl]carbamoyl}1,3-thiazole -2-yl)methyl]carbamate (126)

6, technique

Similar to the general

2-({[2-(1H-1,3-Benzodiazol-2-yl)ethyl][(tert-butoxy)carbonyl]amino}methyl)-1,3-thiazole-4-carboxylic acid (124) (175 mg , 0.36 mmol, 83% purity), (3-fluoropyridin-2-yl)methanamine dihydrochloride (A2) (108 mg, 0.54 mmol), DIPEA (0.25 ml, 1.44 mmol) and HATU ( 206 mg, 0.54 mmol) in DMF (4 mL) gave the title compound (81 mg, 44%) as a colorless solid after purification by basic prep-HPLC.

- 138 039916

1H NMR (DMSO-d ₆ , 500 MHz): δ [ppm] = 12.24 (s, 1H), 8.75 (s, 1H), 8.36 (s, 1H), 8.22 (s, 1H), 7.73-7.66 (m, 1H), 7.50 (s, 1H), 7.46-7.37 (m, 2H), 7.11 (s, 2H), 4.72 (s, 2H), 4.66 (d, J=4.4 Hz, 2H), 3.76 (m, 2H), 3.203.05 (m, 2H), 1.30 (s, 9H ).

HPLC-MS (Method A): [m/z]: 511.10 [M+H]+.

tert-Butyl N-[3-(1H-1,3-benzodiazol-2-yl)propyl]-N-[(4-{[(3-fluoropyridin-2yl)methyl]carbamoyl}-1,3-thiazol- 2-yl)methyl]carbamate (127)

Similar to general procedure 6, 2-({[3-(III-1,3-benzodiazol-2-yl)propyl][(tert-butoxy)carbonyl]amino}methyl)-1,3-thiazole-4-carboxylic acid (125 ) (406 mg, 0.51 mmol, 52% purity), (3-fluoropyridin-2-yl)methanamine dihydrochloride (A2) (152 mg, 0.76 mmol), DIPEA (0.36 ml, 2.04 mmol ) and HATU (291 mg, 0.76 mmol) in DMF (4 mL) gave the title compound (215 mg, 75%) as a white solid after purification by flash column chromatography (kp-NH, gradient elution 70-100% EtOAc/heptane).

1H NMR (DMSO-d ₆ , 500 MHz): δ [ppm] = 12.15 (s, 1H), 8.68 (t, J=5.5 Hz, 1H), 8.36 (s , 1H), 8.22 (s, 1H), 7.69 (t, J=9.3 Hz, 1H), 7.52-7.45 (m, 1H), 7.42-7.34 ( m, 2H), 7.09 (s, 2H), 4.71 (s, 2H), 4.65 (d, J=5.5 Hz, 2H), 3.41 (s, 2H), 2, 81 (t, J=7.3 Hz, 2H), 2.04 (m, 2H), 1.34 (s, 9H).

HPLC-MS (method A): [m/z]: 525.15 [M+H] ⁺ .

tert-Butyl N-[3-(1H-1,3-benzodiazol-2-yl)propyl]-N-[(4-{[(3,5-difluoropyridin-2yl)methyl]carbamoyl}-1,3- thiazol-2-yl)methyl]carbamate (128)

Similar to general procedure 6, 2-({[3-(1H-1,3-benzodiazol-2-yl)propyl][(tert-butoxy)carbonyl]amino}methyl)-1,3-thiazole-4-carboxylic acid (125 ) (0.146 g, 0.351 mmol), (3,5difluoropyridin-2-yl)methanamine dihydrochloride (0.114 g, 0.526 mmol), DIPEA (0.305 ml, 1.753 mmol) and HATU (0.227 g, 0.526 mmol) in DMF (3 ml ) at room temperature for 2 hours gave the title compound (0.071 g, 37%) as a glassy solid after purification by basic prep-HPLC.

1H NMR (DMSO-d6, 500 MHz): δ [ppm] = 12.14 (s, 1H), 8.73-8.66 (m, 1H), 8.47-8.41 (m , 1H), 8.24-8.18 (m, 1H), 7.95-7.87 (m, 1H), 7.52-7.45 (m, 1H), 7.41-7.35 (m, 1H), 7.15-7.05 (m, 2H), 4.70 (s, 2H), 4.61 (d, J=5.7 Hz, 2H), 3.45-3, 38 (m, 2H), 2.85-2.77 (m, 2H), 2.07-1.99 (m, 2H), 1.34 (s, 9H).

HPLC-MS (method A): [m/z]: 543.15 [M+H] ⁺ .

2-({[2-(1H-1,3-Benzodiazol-2-yl)ethyl]amino}methyl)-N-[(3-fluoropyridin-2-yl)methyl]-1,3-thiazole-4carboxamide trihydrochloride (illustrative example #118)

Similar to general procedure 2, tert-butyl N-[2-(1H-1,3-benzodiazol-2-yl)ethyl]-N-[(4-{[(3fluoropyridin-2-yl)methyl]carbamoyl}-1 ,3-thiazol-2-yl)methyl]carbamate (126) (81 mg, 0.159 mmol) and 12M HC (0.53 ml) in MeOH (5 ml) at room temperature for 4 days and then at 40°C within 4 h gave the title compound (49 mg, 58%) as a white solid.

1H NMR (DMSO-d ₆ , 500 MHz): δ [ppm] = 8.81 (s, 1H), 8.41 (s, 1H), 8.36 (dt, J=4.7, 1.4 Hz, 1H), 7.78 (dd, J=6.1, 3.1 Hz, 2H), 7.72 (dd, J=10.0, 8.4, 1.2 Hz, 1H ), 7.53 (dd, J=6.0, 3.1 Hz, 2H), 7.40 (dd, J=8.4, 4.3 Hz, 1H), 4.73 (s, 2H) , 4.69 (d, J=5.1 Hz, 2H), 3.79 (brs, 2H), 3.73 (brs, 2H).

HPLC-MS (Method D): [m/z]: 411.1 [M+H] ⁺ .

2-({[3-(1H-1,3-Benzodiazol-2-yl)propyl]amino}methyl)-N-[(3-fluoropyridin-2-yl)methyl]-1,3-thiazole4-carboxamide trihydrochloride (illustrative example #119)

- 139 039916

Similar to general procedure 2, 12M HC (0.635 ml) was added to a solution of tert-butyl ^[3-(1^1,3benzodiazol-2-yl)propyl]-N-[(4-{[(3-fluoropyridin- 2-yl)methyl]carbamoyl }-1,3-thiazol-2yl)methyl]carbamate (127) (215 mg, 0.381 mmol) in MeOH (5 ml) and the resulting mixture was stirred for 16 h. 0.635 ml, 7.623 mmol) and the resulting mixture was stirred for an additional 20 hours. The reaction mixture was evaporated in vacuo to give the title compound (139 mg, 68%) as a white solid.

1H NMR (DMSO-66, 500 MHz): δ [ppm] = 9.96 (b.s, 1H), 8.78 (b.s, 1H), 8.40 (s, 1H), 8.38 (d, J=4.4 Hz, 1H), 7.80-7.76 (m, 2H), 7.73 (t, J=8.1 Hz, 1H), 7.55-7 .51 (m, 2H), 7.44-7.39 (m, 1H), 4.70 (d, J=4.6 Hz, 2H), 4.64 (s, 2H), 3.35 ( t, J=7.2 Hz, 2H), 3.23 (t, J=7.4 Hz,

2H), 2.39-2.33 (m, 2H).

HPLC-MS (Method D): [m/z]: 425.2 [M+H] ⁺ .

2-({[3-(1H-1,3-Benzodiazol-2-yl)propyl]amino}methyl)-N-[(3,5-difluoropyridin-2-yl)methyl]-1,3thiazol-4- carboxamide dihydrochloride (illustrative example No. 124)

Similar to general procedure 2, 12M HC (0.524 ml, 6.28 mmol) was added to a solution of tert-butyl N[3-(1H-1,3-benzodiazol-2-yl)propyl]-N-[(4-{[ (3,5-difluoropyridin-2-yl)methyl]carbamoyl}-1,3-thiazol-2yl)methyl]carbamate (128) (0.071 g, 0.131 mmol) in MeOH (3 ml) at 45°C for 4 h to give the title compound (0.036g, 53%) as a white solid.

¹ H NMR (Methanol-b ₄ , 500 MHz): δ [ppm] = 8.37 (s, 1H), 8.35-8.30 (m, 1H), 7.81-7.76 (m, 2H), 7.66-7.57 (m, 3H), 4.79 (s, 2H), 4.74 (s, 2H), 3.45-3.36 (m, 4H), 2.51-2.41 (m, 2H).

HPLC-MS (Method D): [m/z]: 443.2 [M+H]+.

The above general scheme 8:

general technique 10:

3-bromo-N-(2-nitrophenyl)propanamide (129)

3-Bromopropanoyl chloride (1.59 ml, 15.75 mmol) was added dropwise to an ice-cold solution of 2-nitroaniline (2.18 g, 15.75 mmol) and TEA (2.63 ml, 18.9 mmol) in toluene (50 ml) and the resulting mixture was stirred for 2 h. The reaction mixture was evaporated in vacuo and the resulting residue was triturated in water (10 ml) to give a brown precipitate which was separated by filtration. Purification by flash column chromatography (eluting with a 0-10% EtOAc/heptane gradient) gave the title compound (0.988 g, 23%) as a yellow crystalline solid.

1H NMR (CDCl3, 250 MHz): δ [ppm] = 10.45 (s, 1H), 8.81 (dd, J=8.5, 1.2 Hz, 1H), 8.25 ( dd, J=8.5, 1.6 Hz, 1H), 7.78-7.61 (m, 1H), 7.24 (dd, J=8.5, 7.3, 1.3 Hz, 1H), 3.74 (t, J=6.5 Hz, 2H), 3.11 (t, J=6.5 Hz, 2H).

HPLC-MS (method A): [m/z]: 272.95/274.90 [M+H] ⁺ .

3-Bromo-N-(4-fluoro-2-nitrophenyl)-propanamide (130)

Similar to general procedure 10, 3-bromopropanoyl chloride (2.29 ml, 23.06 mmol), 4-fluoro-2-nitroaniline (3 g, 19.22 mmol) and TEA (3.124 ml, 23.06 mmol) in toluene (35 ml) at room temperature for 40 h gave the title compound (3.04 g, 42%) as a yellow solid after purification by flash column chromatography (eluting with a 0-40% EtOAc/heptane gradient).

1H NMR (DMSO-66, 500 MHz): δ [ppm] = 10.38 (s, 1H), 7.91 (dd, J=8.6, 2.5 Hz, 1H), 7, 65-7.60 (m, 2H), 3.68 (t, J=6.4 Hz, 2H), 2.97 (t, J=6.4 Hz, 2H).

- 140 039916

HPLC-MS (method A): [m/z]: 290.75/292.75 [M+H]+. 4-Bromo-N-(2-nitrophenyl)butanamide (131)

Similar to general procedure 10, 4-bromobutanoyl chloride (1.56 ml, 13.48 mmol), 2-nitroaniline (1.55 g, 11.24 mmol) and TEA (1.566 ml, 11.2 mmol) in toluene (25 mL) at room temperature for 16 h gave the title compound (2.35 g, 41%) as a yellow oil after purification by flash column chromatography (eluting with a 0-40% EtOAc/heptane gradient).

¹ H NMR (DMSO-06, 500 MHz): δ [ppm] = 10.33 (s, 1H), 7.93 (dd, J=8.2, 1.4 Hz, 1H), 7 .72-7.65 (m, 1H), 7.58 (dd, J=8.1, 1.2 Hz, 1H), 7.38-7.34 (m, 1H), 3.58 (t , J=6.6 Hz, 2H), 2.51 (t, J=6.6 Hz, 2H, overlap with DMSO signals), 2.13-2.06 (m, 2H).

HPLC-MS (method A): [m/z]: 288.75 [M+H] ⁺ .

General Procedure 11: Ethyl 2-[2-({2-[(2-nitrophenyl)carbamoyl]ethyl}amino)ethyl]-1,3-thiazole-4carboxylate(132)

To a solution of 3-bromo-N-(2-nitrophenyl)propanamide (129) (1.04 g, 3.8 mmol) in DMF (10 ml) was added dropwise over 20 min to a mixture of ethyl 2-(2-aminoethyl )-1,3-thiazole-4-carboxylate hydrochloride (1 g, 3.8 mmol, 90% purity) and Na ₂ CO ₃ (0.48 g, 4.56 mmol) in DMF (30 ml). The reaction mixture was stirred for 16 hours at room temperature. Water (10 ml) was added and the resulting mixture was extracted with ethyl acetate (3x20 ml). The combined organic extracts were washed with saturated sodium chloride solution (10 ml), dried (MgSO ₄ ), filtered and evaporated in vacuo to give the crude title compound (1.5 g, 70%, 70% pure) which was used without purification.

HPLC-MS (method A): [m/z]: 393.1 [M+H] ⁺ .

Ethyl 2-[2-({2-[(4-fluoro-2-nitrophenyl)carbamoyl]ethyl}amino)ethyl]-1,3-thiazole-4-carboxylate (133)

Similar to general procedure 11, 3-bromo-N-(4-fluoro-2-nitrophenyl) propanamide (130) (1 g, 2.68 mmol), ethyl 2-(2-aminoethyl)-1,3-thiazole-4 -carboxylate hydrochloride (0.634 g, 2.68 mmol) and Na ₂ CO ₃ (0.426 g, 4.02 mmol) in DMF (10 mL) at room temperature over 24 hours gave the crude title compound (2.26 g , 80%, 39% pure) which was used without purification.

HPLC-MS (method A): [m/z]: 411 [M+H] ⁺ .

Ethyl 2-[2-({3-[(2-nitrophenyl)carbamoyl]propyl}amino)ethyl]-1,3-thiazole-4-carboxylate (134)

Similar to general procedure 11, 4-bromo-N-(2-nitrophenyl)butanamide (131) (2.35 g, 4.65 mmol), ethyl 2-(2-aminoethyl)-1,3-thiazole-4-carboxylate hydrochloride (1.10 g, 4.66 mmol), Na ₂ CO ₃ (0.74 g, 6.98 mmol) and DMF (25 ml) at room temperature for 16 h gave the crude title compound (3, 0 g, quant) as a yellow oil, which was used in the next step without purification.

HPLC-MS (method A): [m/z]: 407 [M+H] ⁺ .

Ethyl 2-(2-{[(tert-butoxy)carbonyl]({2-[(2-nitrophenyl)carbamoyl]ethyl})amino}ethyl)-1,3-thiazole4-carboxylate (135)

- 141 039916

Similar to general procedure 4, ethyl 2-[2-({2-[(2-nitrophenyl)carbamoyl]ethyl}amino)ethyl]-1,3thiazole-4-carboxylate (132) (1.3 g, 1.99 mmol , 60% pure), Boc ₂ O (477 mg, 2.19 mmol) and TEA (413 μl,

2.9 mmol) in THF (50 ml) was stirred at room temperature for 16 h. Additional Boc ₂ O (477 mg, 2.19 mmol) and TEA (413 μl, 2.98 mmol) were added and the resulting mixture stirred for another 4 h. The reaction mixture was evaporated in vacuo, the resulting residue was dissolved in EtOAc

(10 ml) and washed with water (3x5 ml). The organic layer was dried (MgSO ₄ ), filtered and evaporated in vacuo. Purification by flash column chromatography (gradient elution 10-100%

EtOAc/heptane) gave the title compound (132 mg, 12%) as a yellow oil.

HPLC-MS (Method A): [m/z]: 493.15 [M+H]+.

Ethyl 2-(2-{[(tert-butoxy)carbonyl]({2-[(4-fluoro-2-nitrophenyl)carbamoyl]ethyl})amino}ethyl)-1,3thiazole-4-carboxylate (136)

Similar to general procedure 4, 2-[2-({2-[(4-fluoro-2-nitrophenyl)carbamoyl]ethyl}amino)ethyl]-1,3thiazole-4-carboxylate (133) (2.26 g, 2.145 mmol), Boc ₂ O (1.87 g, 8.58 mmol) and TEA (0.848 ml, 6.43 mmol) in THF (60 ml) at room temperature for 16 h gave the title compound (0.43 g, 38%) as a yellow oil after purification by flash column chromatography (eluting with a 10-100% EtOAc/heptane gradient).

¹ H NMR (DMSO-d ₆ , 500 MHz): δ [ppm] = 10.30 (s, 1H), 8.41 (s, 1H), 7.89 (d, J=8.5 Hz, 1H), 7.62 (m, 2H), 4.29 (q, J=7.1 Hz, 2H), 3.54 (m, 2H), 3.41 (t, J=7.1 Hz, 2H), 3.22 (t, J=7.0 Hz, 2H), 2.58 (m 2H), 1.351.27 (m, 12H).

HPLC-MS (method A): [m/z]: 511.1 [M+H]+.

Ethyl 2-(2-{[(tert-butoxy)carbonyl]({3-[(2-nitrophenyl)carbamoyl]propyl})amino}ethyl)-1,3thiazole-4-carboxylate (137)

about

As in general procedure 4, ethyl 2-[2-({3-[(2-nitrophenyl)carbamoyl]propyl}amino)ethyl]-1,3thiazole-4-carboxylate (134) (3.0 g, 5.32 mmol, 72% pure), Boc ₂ O (2.44 g, 11.17 mmol) and TEA (2.10 ml, 15.96 mmol) in THF (50 ml) were stirred at room temperature for 24 h. additional Boc2O (2.32 g, 10.64 mmol) and TEA (0.7 ml, 5.32 mmol) and the reaction mixture was stirred at room temperature for 96 h to give the title compound (0.287 g, 10 %) as a yellow oil after purification by reverse phase column chromatography (elution with a gradient of 0-100% MeCN/water).

¹ H NMR (DMSO-J6, 500 MHz): δ [ppm] = 10.23 (s, 1H), 8.40 (s, 1H), 7.93 (dd, J=8.2, 1.4 Hz, 1H), 7.717.66 (m, 1H), 7.62 (dd, J=8.1, 1.4 Hz, 1H), 7.37-7.32 (m, 1H), 4.28 (q, J=7.1 Hz, 2H), 3.53 (t, J=6.9 Hz, 2H), 3.25-3.15 (m, 4H), 2.32 (t , J=7.4 Hz, 2H), 1.81-1.72 (m, 2H), 1.40-1.29 (breadth m, 9H), 1.28 (t, J=7.1 Hz, 3H).

HPLC-MS (method A): [m/z]: 507.1 [M+H] ⁺ .

Ethyl 2-(2-{[2-(1H-1,3-benzodiazol-2-yl)ethyl][(tert-butoxy)carbonyl]amino}ethyl)-1,3-thiazole-4carboxylate(138)

- 142 039916

Suspension of ethyl 2-(2-{[(tert-butoxy)carbonyl]({2-[(2-nitrophenyl)carbamoyl]ethyl})amino}ethyl)1,3-thiazole-4-carboxylate (135) (175 mg , 0.36 mmol) and iron powder (238 mg, 4.26 mmol) in AcOH were heated at 80°C for 1 h. The reaction mixture was cooled to room temperature, diluted with DCM (10 ml) and neutralized with saturated NaHCO ₃ solution . The aqueous phase was extracted with dichloromethane (3x10 ml), dried (Na ₂ SO ₄ ), filtered and evaporated in vacuo to give the title compound (121 mg, 76%) as a pale yellow oil.

HPLC-MS (Method A): [m/z]: 445.15 [M+H]+.

Ethyl 2-(2-{[(tert-butoxy)carbonyl] [2-(5-fluoro-1H-1,3-benzodiazol-2-yl)ethyl]amino }ethyl)-1,3thiazole-4-carboxylate ( 139)

A suspension of ethyl 2-(2-{[(tert-butoxy)carbonyl]({2-[(4-fluoro-2-nitrophenyl)carbamoyl]ethyl})amino}ethyl)-1,3-thiazole-4-carboxylate ( 136) (0.43 g, 0.825 mmol) and iron powder (0.533 g, 9.905 mmol) in AcOH (40 ml) were heated at 80°C for 2 h. The reaction mixture was cooled to room temperature and neutralized by slowly adding saturated Na ₂ CO ₃ . The resulting mixture was extracted with dichloromethane (4x40 ml) and the combined organic extracts were dried (MgSO ₄ ), filtered and evaporated in vacuo to give the title compound as an off-white glassy solid (0.445 g, quant.).

¹ H NMR (DMSO-d6, 500 MHz): δ [ppm] = 12.34 (s, 1H), 8.40 (s, 1H), 7.27 (s, 2H), 6.96 (s, 1H), 4.28 (q, J=7.1 Hz, 2H), 3.59 (t, J=7.1 Hz, 2H), 3.55 (br s, 2H), 3 .22 (bds, 2H), 3.00 (bds, 2H), 1.34-1.20 (m, 12H).

HPLC-MS (method A): [m/z]: 463.1 [M+H] ⁺ .

Ethyl 2-(2-{[3-(1H-1,3-benzodiazol-2-yl)propyl][(tert-butoxy)carbonyl]amino}ethyl)-1,3-thiazole-4carboxylate(140) o / ^Vos l J

Η

Iron powder (0.368 g, 6.595 mmol) was added to ethyl 2-(2-{[(tert-butoxy)carbonyl]({3-[(2nitrophenyl)carbamoyl]propyl})amino}ethyl)-1,3-thiazole- 4-carboxylate (137) (0.287 g, 0.55 mmol, 97% pure) in AcOH (10 ml). The reaction mixture was stirred at 80° C. for 1 hour. The reaction mixture was allowed to cool to room temperature. Was added water (50 ml), then Na ₂ CO ₃ to pH ~9. The aqueous layer was extracted with dichloromethane (4x50 ml). The combined organic layers were dried (MgSO ₄ ), filtered and the solvent was evaporated to give the title compound (0.291 g, quant) as a pale orange oil.

1H NMR (DMSO-d ₆ , 500 MHz): δ [ppm] = 12.15 (s, 1H), 8.39 (s, 1H), 7.53-7.47 (m, 1H) , 7.43-7.35(m, 1H), 7.14-7.06(m, 2H), 4.28(q, J=7.1Hz, 2H), 3.58-3.51 (m, 2H), 3.26-3.21 (m, 4H), 2.77 (t, J=7.6 Hz, 2H), 2.01-1.91 (m, 2H), 1, 30 (s, 9H), 1.28 (t, J=7.1 Hz, 3H).

HPLC-MS (method A): [m/z]: 459.1 [M+H] ⁺ .

2-(2-{[2-(1H-1,3-Benzodiazol-2-yl)ethyl][(tert-butoxy)carbonyl]amino}ethyl)-1,3-thiazole-4carboxylic acid (141) o

| ^but . Dos ⁴ Ν ύ

Similar to general procedure 5, ethyl 2-(2-{[2-(1H-1,3-benzodiazol-2-yl)ethyl][(tert-butoxy)carbonyl]amino}ethyl)-1,3-thiazole-4-carboxylate (138) (156 mg, 0.35 mmol) and LiOH (33 mg, 1.35 mmol) in THF/water (5 mL/1 mL) gave the title compound (100 mg, 68%) as a brownish solid after acidification with AcOH, extraction with 3:1 THF/EtOAc (3x10 ml), drying (MgSO ₄ ), filtering and evaporating in vacuo.

HPLC-MS (method A): [m/z]: 417.1 [M+H]+.

- 143 039916

2-(2-{[(tert-Butoxy)carbonyl][2-(5-fluoro-1H-1,3-benzodiazol-2-yl)ethyl]amino }ethyl)-1,3-thiazole-4carboxylic acid ( 142)

Similar to general procedure 5, ethyl 2-(2-{[(tert-butoxy)carbonyl][2-(5-fluoro-1H-1,3-benzodiazol2-yl)ethyl]amino}ethyl)-1,3-thiazole -4-carboxylate (139) (380 mg, 0.83 mmol) and LiOH (59 mg, 2.48 mmol) in THF/water (45 mL/15 mL) gave the title compound (319 mg, 82% , 92% pure) as a white solid.

HPLC-MS (method A): [m/z]: 435.05 [M+H]+.

2-(2-{[3-(1H-1,3-Benzodiazol-2-yl)propyl][(tert-butoxy)carbonyl]amino}ethyl)-1,3-thiazole-4carboxylic acid (143)

Similar to general procedure 5, ethyl 2-(2-{[3-(1H-1,3-benzodiazol-2-yl)propyl][(tert-butoxy)carbonyl]amino}ethyl)-1,3-thiazole-4-carboxylate (140) (291 mg, 0.550 mmol, 87% pure) and LiOH (39 mg, 1.649 mmol) in THF/water (25 ml/10 ml) at room temperature for 24 h gave the title compound (219 mg , 72%) as a glassy solid.

¹ H NMR (Acetone-de, 500 MHz): δ [ppm] = 7.87 (s, 1H), 7.06-6.96 (m, 2H), 6.70-6.63 ( m, 2H), 3.90-3.85 (m, 2H), 3.10 (t, J=6.8 Hz, 2H), 2.76 (t, J=7.0 Hz, 2H), 2.34 (t, J=7.4 Hz, 2H), 1.57-1.49 (m, 2H), 0.85 (s, 9H).

HPLC-MS (method A): [m/z]: 431.1 [M+H] ⁺ .

tert-Butyl N-[2-(1H-1,3-benzodiazol-2-yl)ethyl]-N-[2-(4-{[(3,5-difluoropyridin-2 yl)methyl]carbamoyl}-1 ,3-thiazol-2-yl)ethyl]carbamate (144)

Similar to the general procedure 6, butoxy)carbonyl]amino}ethyl)-1,3-thiazole-4-carboxylic

2-(2-{[2-(1H-1,3-benzodiazol-2-yl)ethyl][(tert acid (141) (100mg, 0.24mmol), (3,5difluoropyridin-2-yl)methanamine dihydrochloride (78 mg, 0.36 mmol), DIPEA (0.21 ml, 1.2 mmol) and HATU (137 mg, 0.36 mmol) in DMF (3 ml) gave the title compound (77 mg, 59 %) as a white solid after purification by basic prep-HPLC.

1H NMR (Methanol-d4, 500 MHz): δ [ppm] = 8.30 (br s, 1H), 8.09 (s, 1H), 7.55 (br s, 1H), 7.48 (s, 2H), 7.20 (d, J=3.3 Hz, 2H), 4.82 (s, 2H), 4.73 (s, 2H), 3.70 (m, 4H ), 3.08 (m, 2H), 1.16 (s, 9H).

HPLC-MS (method A): [m/z]: 543.15 [M+H] ⁺ .

tert-Butyl N-[2-(5-fluoro-1H-1,3-benzodiazol-2-yl)ethyl]-N-[2-(4-{[(3-fluoropyridin-2yl)methyl]carbamoyl}- 1,3-thiazol-2-yl)ethyl]carbamate (145)

Similar to general procedure 6, 2-(2-{[(tert-butoxy)carbonyl][2-(5-fluoro-1H-1,3-benzodiazol-2yl)ethyl]amino}ethyl)-1,3-thiazol- 4-carboxylic acid (142) (160 mg, 0.34 mmol, 92% pure), (3fluoropyridin-2-yl)methanamine dihydrochloride (A2) (101 mg, 0.51 mmol), DIPEA (0.18 ml, 1.01 mmol) and HATU (192 mg, 0.51 mmol) in DMF (4 ml) gave the title compound (112 mg, 61%) as a white solid after purification by flash column chromatography (gradient elution 0-50% MeOH/EtOAc).

1H NMR (DMSO-d ₆ , 500 MHz): δ [ppm] = 12.33 (br s, 1H), 8.66 (br s, 1H), 8.36 (br s, 1H), 8.17 (brs, 1H), 7.72-7.66 (m, 1H), 7.53-7.36 (m, 2H), 7.33-7.21 (m, 1H), 7.00-6.91 (m, 1H), 4.66 (d, J=4.5

- 144 039916

Hz, 2H), 3.63 (t, J=7.0 Hz, 2H), 3.58 (t, J=6.9 Hz, 2H), 3.24 (t, J=6.9 Hz, 2H), 3.02 (m, 2H), 1.35-1.18 (m, 9H).

HPLC-MS (method A): [m/z]: 543.1 [M+H]+.

tert-Butyl X-[2-(4-{[(3,5-difluoropyridin-2-yl)methyl]carbamoyl}-1,3-thiazol-2-yl)ethyl]-X-[2-(5fluoro- 1H-1,3-benzodiazol-2-yl)ethyl]carbamate( 146)

Similar to the general procedure 6, 2-(2-{[(tert-butoxy)karbonyl][2-(5-fluoro-1H-1,3-benzodiazol-2yl)ethyl]amino}ethyl)-1,3-thiazol- 4-carboxylic acid (142) (12 mg, 0.28 mmol), (3,5-difluoropyridin-2yl)methanamine dihydrochloride (90 mg, 0.41 mmol), DIPEA (0.24 ml, 1.38 mmol) and HATU (158 mg, 0.41 mmol) in DMF (3 ml) gave the title compound (72 mg, 47%) as a colorless glassy solid after purification by basic prep-HPLC.

¹ H NMR (DMSO-d ₆ , 500 MHz): δ [ppm] = 12.33 (b.s, 1H), 8.68 (b.s, 1H), 8.44 (b.s , 1H), 8.16 (brs, 1H), 7.90 (d, J=11.0 Hz, 1H), 7.27 (brs, 2H), 6.96 (brs, 1H), 4.62 (d, J=5.7 Hz, 2H), 3.653.57 (m, 4H), 3.22 (m, 2H), 3.01 (brs, 2H), 1, 24 (m, 9H).

HPLC-MS (Method A): [m/z]: 561.15 [M+H]+.

tert-Butyl N-[3-(1H-1,3-benzodiazol-2-yl)propyl]-N-[2-(4-{[(3-fluoropyridin-2yl)methyl]carbamoyl}-1,3- thiazol-2-yl)ethyl] carbamate (147)

F about

H

Similar to general procedure 6, 2-(2-{[3-(1H-1,3-benzodiazol-2-yl)propyl][(tert-butoxy)carbonyl]amino}ethyl)-1,3-thiazole-4-carboxylic acid (143) (219 mg, 0.39 mmol, 78% pure), (3-fluoropyridin-2-yl)methanamine dihydrochloride (A2) (118 mg, 0.59 mmol), DIPEA (0.346 ml, 1.98 mmol ) and HATU (226 mg, 0.59 mmol) in DMF (3 mL) at room temperature for 2 h gave the title compound (111 mg, 52%) as a colorless oil after purification by basic pre-HPLC.

1H NMR (DMSO-H6, 500 MHz): δ [ppm] = 12.15 (s, 1H), 8.66 (br s, 1H), 8.36 (d, J=4.3 Hz, 1H), 8.16 (s, 1H), 7.73-7.63 (m, 1H), 7.52-7.45 (m, 1H), 7.45-7.34 (m, 2H), 7.18-7.01 (m, 2H), 4.65 (d, J=5.5 Hz, 2H), 3.59 (t, J=7.0 Hz, 2H), 3, 29-3.23 (m, 4H), 2.78 (t, J=7.5 Hz, 2H), 2.05-1.92 (m, 2H), 1.30 (s, 9H).

HPLC-MS (method A): [m/z]: 539.15 [M+H]+.

2-(2-{[2-(1H-1,3-Benzodiazol-2-yl)ethyl]amino}ethyl)-X-[(3,5-difluoropyridin-2-yl)methyl]-1,3thiazol- 4-carboxamide trihydrochloride (illustrative example No. 116)

Similar to general procedure 2, 12M HC (0.378 mL, 4.541 mmol) and t-butyl ^[2-(1^1,3benzodiazol-2-yl)ethyl]-X-[2-(4-{[(3.5 -difluoropyridin-2-yl)methyl]carbamoyl}-1,3-thiazol-2yl)ethyl]carbamate (144) (77 mg, 0.142 mmol) in MeOH (3 ml) at room temperature for 5 h and at 40° C gave the title compound (60 mg, 73%) as a yellow solid over 20 hours.

1H NMR (Methanol-b ₄ , 500 MHz): δ [ppm] = 8.29 (d, J=2.3 Hz, 1H), 8.23 (s, 1H), 7.79 (dd , J=6.1, 3.2 Hz, 2H), 7.64-7.59 (m, 3H), 4.77 (s, 2H), 3.78 (s, 4H), 3.72 ( t, J=6.4 Hz, 2H), 3.59 (d, J=5.9 Hz, 2H).

HPLC-MS (Method D): [m/z]: 443.1 [M+H]+.

2-(2-{[2-(5-Fluoro-1H-1,3-benzodiazol-2-yl)ethyl]amino}ethyl)-N-[(3-fluoropyridin-2-yl)methyl]-1, 3thiazole-4-carboxamide trihydrochloride (illustrative example No. 120)

- 145 039916

Similar to general procedure 2, 12M HC (0.344 ml, 4.128 mmol) and N--[2-(5-φτορ-1Η-1,3-benzodiazol-2-yl)ethyl]-N-[2-(4-{[( 3-fluoropyridin-2-yl)methyl]carbamoyl}-1,3-thiazol-2-yl)ethyl]carbamate (145) (112 mg, 0.206 mmol) in MeOH (4 ml) at room temperature for 16 h, followed by

addition of 12M HC (0.344 mL, 4.128 mmol) at room temperature for an additional 20 h and at 40° C. for 3 h gave the title compound (80 mg, 67%) as a white solid.

1H NMR (Methanol-b4, 500 MHz): δ [ppm] = 8.57 (d, J=5.4 Hz, 1H), 8.28 (t, J=8.8 Hz, 1H) , 8.25 (s, 1H), 7.89 (dt, J=9.5, 5.0 Hz, 1H), 7.81 (dd, J=9.0, 4.3 Hz, 1H), 7.58 (dd, J=8.1, 2.3 Hz, 1H), 7.42 (td, J=9.3, 2.4 Hz, 1H), 4.97 (s, 2H), 3 .85-3.79 (m, 4H), 3.75 (t, J=6.3 Hz, 2H), 3.62 (t, J=6.3 Hz, 2H).

HPLC-MS (Method D): [m/z]: 443.2 [M+H]+.

N-[(3,5-difluoropyridin-2-yl)methyl]-2-(2-{[2-(5-fluoro-1H-1,3-benzodiazol-2-yl)ethyl]amino}ethyl)1 ,3-thiazole-4-carboxamide trihydrochloride (illustrative example No. 123)

Similar to general procedure 2, 12M HC (0.257 ml, 3.08 mmol) and tert-butyl N-[2-(4-{[(3,5difluoropyridin-2-yl)methyl]carbamoyl}-1,3-thiazol- 2-yl)ethyl]-N-[2-(5-fluoro-1H-1,3-benzodiazol-2yl)ethyl]carbamate (146) (72 mg, 0.128 mmol) in MeOH (3 ml) at room temperature in over 16 hours, followed by addition of 12M HC (0.257 mL, 3.08 mmol) and stirring of the resulting mixture at 45° C. for 4 hours, gave the title compound (28 mg, 38%) as a white solid.

1H NMR (Methanol-a ₄ , 500 MHz): δ [ppm] = 8.28 (d, J=2.3 Hz, 1H), 8.23 (s, 1H), 7.79 (dd , J=9.0, 4.3 Hz, 1H), 7.60 (m, 1H), 7.56 (dd, J=8.1, 2.3 Hz, 1H), 7.40 (td, J=9.3, 2.4 Hz, 1H), 4.76 (s, 2H), 3.77 (m, 4H), 3.73 (t, J=6.4 Hz, 2H), 3, 60 (t, J=6.4 Hz, 2H).

HPLC-MS (Method D): [m/z]: 461.2 [M+H] ⁺ .

2-(2-{[3 -(1H-1,3-Benzodiazol-2-yl)propyl]amino}ethyl)-N-[(3-fluoropyridin-2-yl)methyl]-1,3-thiazol4- carboxamide trihydrochloride (illustrative example No. 125)

Similar to general procedure 2, 12M HC (0.405 mL, 4.864 mmol) and t-butyl ^[3-(1^1,3benzodiazol-2-yl)propyl]-N-[2-(4-{[(3-fluoropyridine -2-yl)methyl]carbamoyl}-1,3-thiazol-2yl)ethyl]carbamate (147) (131 mg, 0.243 mmol) in MeOH (3 mL) at 40°C for 16 h gave the title compound (48 mg, 35%) as a white solid.

1H NMR (Methanol-a ₄ , 500 MHz): δ [ppm] = 8.48 (d, J=5.1 Hz, 1H), 8.23 (s, 1H), 8.07-8 .00 (m, 1H), 7.807.76 (m, 2H), 7.74-7.66 (m, 1H), 7.63-7.58 (m, 2H), 4.91 (s, 2H ), 3.65 (t, J=6.4 Hz, 2H), 3.57 (t, J=6.6 Hz, 2H), 3.41-3.32 (m, 4H), 2.49 -2.40 (m, 2H).

HPLC-MS (Method D): [m/z]: 439.2 [M+H] ⁺ .

General Method 11:

tert-butyl 3-carbamothioylpyrrolidine-1-carboxylate (148)

Lawson's reagent (0.42 g, 1.03 mmol) was added in one portion to a solution of tert-butyl 3-carbamoylpyrrolidine-1-carboxylate (0.4 g, 1.87 mmol) in DCM (5 mL) and the reaction mixture was stirred at room temperature for 2 hours. The reaction mixture was transferred directly onto silica gel and purified by flash column chromatography (eluting with a 0-100% EtOAc/heptane gradient) to give the title compound (0.36 g, 79%) as off-white solid.

1H NMR (CDCl3, 500 MHz): δ [ppm] = 7.46 (d, J=8.6 Hz, 1H), 7.01 (br s, 1H), 3.75-3, 66 (m, 1H), 3.653.54 (m, 1H), 3.59 (dd, 11 and 8.1 Hz, 1H), 3.37 (kv, J=8.7 Hz, 1H), 3, 27 (n, J=7.8 Hz, 1H), 2.24-2.12 (m, 3H), 1.46

- 146 039916 (s, 9H).

HPLC-MS (Method A): [m/z]: 252.95 [M+Na]+.

Ethyl 2-{1-[(tert-butoxy)carbonyl]pyrrolidin-3-yl}-1,3-thiazole-4-carboxylate (149) o

Similar to general procedure 1, ethyl 3-bromo-2-oxopropanoate (2.5 ml, 19.75 mmol), tert-butyl 3-carbamothioylpyrrolidine-1-carboxylate (148) (3.96 g, 17.19 mmol) and CaCO ₃ (0.87 g, 8.73 mmol) in EtOH (50 mL) at room temperature over 21 h gave the title compound (2.84 g, 51.9%) as a yellow oil which solidified on standing, after purification by flash column chromatography (eluting with a gradient of 0-70% EtOAc/heptane).

¹ H NMR (CDCl ₃ , 250 MHz): δ [ppm] = 8.08 (s, 1H), 4.41 (q, J=7.1 Hz, 2H), 3.96-3, 76 (m, 2H), 3.73-3.51 (m, 2H), 3.51-3.35 (m, 1H), 2.35-2.44 (m, 1H), 2.28- 2.14 (m, 1H), 1.46 (s, 9H), 1.39 (t, J=7.1 Hz, 3H).

HPLC-MS (method A): [m/z]: 349.05 [M+Na]+.

2-{1-[(tert-Butoxy)carbonyl]pyrrolidin-3-yl}-1,3-thiazole-4-carboxylic acid (150) o

HO^^y^QNBoc

Similar to general procedure 5, LiOH (1.04 g, 43.5 mmol) and ethyl 2-{1-[(tert-butoxy)carbonyl]pyrrolidin-3-yl}-1,3-thiazole-4-carboxylate (149) ( 2.84 g, 8.7 mmol) in THF (30 ml)/water (30 ml) gave the title compound (2.48 g, 93.6%) as a yellow solid. The resulting compound was used in the next step without further purification.

1H NMR (DMSO-d6, 250 MHz): δ [ppm] = 12.94 (s, 1H), 8.37 (s, 1H), 3.94-3.78 (m, 1H), 3.78-3.68 (m, 1H), 3.49-3.39 (m, 2H), 2.44-2.24 (m, 1H), 2.19-1.98 (m, 1H ), 1.41 (s, 9H).

The CH signal overlapped with the H ₂ O peak.

HPLC-MS (Method A): [m/z]: 321.05 [M+Na]+ and 297.1 [M+H] ⁺ .

tert-Butyl 3-(4-{[(3-fluoropyridin-2-yl)methyl]carbamoyl}-1,3-thiazol-2-yl)pyrrolidine-1carboxylate(151)

Similarly, using the general procedure 6, 2-{1-[(tert-butoxy)carbonyl]pyrrolidin-3yl}-1,3-thiazole-4-carboxylic acid (150) (1 g, 3.35 mmol), ( 3-fluoropyridin-2-yl)methanamine dihydrochloride (A2) (1 g, 5.03 mmol), DIPEA (1.93 ml, 11.06 mmol), HATU (1.9 g, 5.03 mmol) in DMF (14 mL) gave the title compound (1.27 g, 86%) as a yellow oil after purification by flash column chromatography (elution with a 0-100% EtOAc/heptane gradient), followed by repurification by flash column chromatography (elution with a 0-10% MeOH/DCM gradient).

1H NMR (DMSO-d6, 250 MHz): δ [ppm] = 8.71 (t, J=5.6 Hz, 1H), 8.38 (dt, J=4.7, 1.4 Hz, 1H), 8.20 (s, 1H), 7.70 (ddd, J=10.2, 8.3, 1.3 Hz, 1H), 7.40 (dt, J=8.5, 4.4 Hz, 1H), 4.66 (dd, J=5.7, 1.5 Hz, 2H), 3.94-3.81 (m, 1H), 3.82-3.66 (m , 1H), 3.58-3.40 (m, 2H), 2.43-2.26 (m, 1H), 2.25-2.04 (m, 1H), 1.40 (s, 9H ). The CH signal overlapped with _the H2O peak.

HPLC-MS (method A): [m/z]: 407.1 [M+H] ⁺ .

tert-Butyl 3-{4-[(pyridin-2-ylmethyl)carbamoyl]-1,3-thiazol-2-yl}pyrrolidin-1-carboxylate (152) o

CoVax

Similarly, using the general procedure 6, 2-{1-[(tert-butoxy)carbonyl]pyrrolidin-3yl}-1,3-thiazole-4-carboxylic acid (150) (1 g, 3.35 mmol), pyridine -2-ylmethanamine (0.52 ml, 5.03 mmol), DIPEA (1.17 ml, 6.7 mmol) and HATU (1.9 g, 5.03 mmol) in DMF (14 ml) gave the indicated title compound (1.54 g, 100%) as a yellow solid after purification by flash column chromatography (eluting with a 0-100% EtOAc/heptane gradient).

1H NMR (DMSO-d ₆ , 250 MHz): δ [ppm] = 8.90 (s, 1H), 8.52 (d, J=4.0 Hz, 1H), 8.22 (s , 1H), 7.76 (td, J=7.7, 1.8 Hz, 1H), 7.36-7.21 (m, 2H), 4.58 (d, J=6.0 Hz, 2H), 3.92-3.82 (m, 1H), 3.84-3.71 (m, 1H), 3.68-3.42 (m, 3H), 2.34-2.25 ( m, 1H), 2.27-2.11 (m, 1H).

HPLC-MS (method A): [m/z]: 389.15 [M+H] ⁺ .

N-[(3-Fluoropyridin-2-yl)methyl]-2-(pyrrolidin-3-yl)-1,3-thiazol-4-carboxamide dihydrochloride (153)

2HCI

- 147 039916

Similarly, using General Procedure 2, 12M HC (6 mL) and tert-butyl 3-(4-{[(3fluoropyridin-2-yl)methyl]carbamoyl}-1,3-thiazol-2-yl)pyrrolidin-1 -carboxylate (151) (1.27 g, 2.87 mmol) in MeOH (25 ml) at 40°C for 4 h gave the title compound (1.11 g, 96.7%) as a creamy foam . The compound was used in the next step without further purification.

Ή NMR (Methanol-04, 250 MHz): δ [ppm] = 8.58 (dd, J=5.4, 1.2 Hz, 1H), 8.28-8.18 (m, 2H ), 7.87 (dt, J=8.6, 4.8 Hz, 1H), 4.94 (d, J=1.3 Hz, 2H), 4.25-4.11 (m, 1H) , 3.82 (dd, J=11.8, 4.6 Hz, 1H), 3.72-3.43 (m, 3H), 2.67-2.46 (m, 1H), 2.39 -2.23 (m, 1H).

HPLC-MS (method A): [m/z]: 307.05 [M+H]+.

N-(Pyridin-2-ylmethyl)-2-(pyrrolidin-3-yl)-1,3-thiazole-4-carboxamide dihydrochloride (154)

2HCI

Similarly, using general procedure 2, 12M HC (7 ml) and tert-butyl 3-{4-[(pyridin-2ylmethyl)carbamoyl]-1,3-thiazol-2-yl}pyrrolidine-1-carboxylate (152) (1.54 g, 3.35 mmol) in MeOH (30 mL) at 40° C. for 4 h gave the title compound (1.42 g, 99%) as a pale brown foam. The compound was used in the next step without further purification.

Ή NMR (Methanol-04, 250 MHz): δ [ppm] = 8.77 (d, J=5.9 Hz, 1H), 8.65-8.54 (m, 1H), 8, 25 (s, 1H), 8.12 (d, J=8.1 Hz, 1H), 7.99 (t, J=6.8 Hz, 1H), 4.96 (s, 2H), 4. 17 (ddd, J=11.5, 7.3, 4.3 Hz, 1H), 3.86 (dd, J=11.9, 4.2 Hz, 1H), 3.79-3.41 ( m, 3H), 2.56 (d.sq., J=13.4, 8.3 Hz, 1H), 2.41-2.21 (m, 1H).

HPLC-MS (method A): [m/z]: 289.05 [M-H] ⁺ .

N-[(3-Fluoropyridin-2-yl)methyl]-2-(1-{2-[(2-nitrophenyl)carbamoyl]ethyl}pyrrolidin-3-yl)-1,3thiazole-4-carboxamide (155)

Similar to general procedure 8, N-[(3-fluoropyridin-2-yl)methyl]-2-(pyrrolidin-3-yl)-1,3-thiazol-4-carboxamide dihydrochloride (153) (1.11 g, 2.78 mmol), N-(2-nitrophenyl)prop-2-enamide (D) (0.59 g, 3.06 mmol), DBU (1.41 ml, 9.45 mmol) in MeCN (20 ml) at room temperature temperature for 19 h gave the title compound (1.53 g, 99%) as a yellow oil after work-up.

¹ H NMR (DMSO-06, 500 MHz): δ [ppm] = 10.69 (s, 1H), 8.67 (t, J=5.7 Hz, 1H), 8.38 (dt , J=4.6, 1.4 Hz, 1H), 8.11 (s, 1H), 7.96 (dd, J=8.2, 1.5 Hz, 1H), 7.79 (dd, J=8.2, 1.2 Hz, 1H), 7.73-7.67 (m, 2H), 7.40 (dt, J=8.6, 4.4 Hz, 1H), 7.33 (ddd, J=8.5, 7.4, 1.4 Hz, 1H), 4.63 (d, J=4.6 Hz, 2H), 3.87-3.77 (m, 1H), 2.99 (dd, J=9.2, 7.4 Hz, 1H), 2.88-2.74 (m, 4H), 2.70-2.62 (m, 1H), 2.57 ( td, J=6.7, 1.9 Hz, 2H), 2.43-2.34 (m, 1H), 2.08-1.99 (m, 1H).

HPLC-MS (method A): [m/z]: 499.1 [M+H] ⁺ .

tert-Butyl 3-{4-[(pyridin-2-ylmethyl)carbamoyl]-1,3-thiazol-2-yl}pyrrolidin-1-carboxylate (illustrative example No. 214)

Similar to general procedure 8, N-[(3-fluoropyridin-2-yl)methyl]-2-(1-{2-[(2nitrophenyl)carbamoyl]ethyl}pyrrolidin-3-yl)-1,3-thiazol-4 -carboxamide (155) (1.53 g, 2.78 mmol), AcOH (15 ml) and iron powder (0.62 g, 11.11 mmol) at 80°C for 2 h gave the title compound ( 94 mg, 8%) as a pale brown solid after purification by flash column chromatography (KP-NH, eluting with a 0-25% MeOH/DCM gradient) to give a residue (207 mg) which was further purified by column flash chromatography (KP-NH, eluting with a 0-3% MeOH/DCM gradient).

1H NMR (Methanol ₄ , 500 MHz): δ [ppm] = 8.33 (d, J=4.7 Hz, 1H), 7.94 (s, 1H), 7.58 (pseudo t, J=9.1 Hz, 1H), 7.47 (brs, 2H), 7.40-7.32 (m, 1H), 7.24-7.16 (m, 2H), 4 .75 (d, J=1.4 Hz, 2H), 3.87-3.80 (m, 1H), 3.17-3.11 (m, 2H), 3.11-2.96 (m , 5H), 2.78-2.69 (m, 1H), 2.48-2.38 (m, 1H), 2.18-2.04 (m, 1H).

HPLC-MS (method C): [m/z]: 451.1 [M+H] ⁺ .

2-(1-{2-[(2-Nitrophenyl)carbamoyl]ethyl}pyrrolidin-3-yl)-N-(pyridin-2-ylmethyl)-1,3-thiazole-4carboxamide (156)

- 148 039916

CO\>O ^{N th}

Similar to general procedure 8, DBU (1.52 mL, 10.19 mmol), N-(pyridin-2-ylmethyl)-2(pyrrolidin-3-yl)-1,3-thiazole-4-carboxamide dihydrochloride (154) (84.6%, 1.28 g, 3.0 mmol) and N-(2nitrophenyl)prop-2-enamide (D) (0.63 g, 3.3 mmol) in MeCN (25 ml) at room temperature gave the title compound (1.43 g, 99%) as a yellow oil over 19 hours after purification by flash column chromatography (eluting with a 0-12% MeOH/DCM gradient).

1H NMR (DMSO-d6, 500 MHz): δ [ppm] = 10.67 (s, 1H), 8.83 (t, J=6.0 Hz, 1H), 8.51 (d, J=4.2Hz, 1H), 8.12(s, 1H), 7.96(dd, J=8.2, 1.4Hz, 1H), 7.82(d, J=8.2 Hz, 1H), 7.74 (td, J=7.7, 1.8 Hz, 1H), 7.69 (td, J=8.5, 8.0, 1.5 Hz, 1H), 7 .36-7.24 (m, 3H), 4.55 (d, J=6.1 Hz, 2H), 3.87-3.78 (m, 1H), 3.04-2.98 (m , 1H), 2.89-2.74 (m, 4H), 2.71-2.65 (m, 1H), 2.57 (td, J=6.7, 1.7 Hz, 2H), 2.44-2.33 (m, 1H), 2.09-2.01 (m, 1H).

HPLC-MS (method A): [m/z]: 481.35 [M+H]+.

2-{1-[2-(1H-1,3-Benzodiazol-2-yl)ethyl]pyrrolidin-3-yl}-N-(pyridin-2-ylmethyl)-1,3-thiazole-4carboxamide (illustrative example No. 215)

Similar to general procedure 8, 2-(1-{2-[(2-nitrophenyl)carbamoyl]ethyl}pyrrolidin-3-yl)-N(pyridin-2-ylmethyl)-1,3-thiazole-4-carboxamide (156 ) (1.43 g, 2.77 mmol) and iron powder (0.62 g, 11.07 mmol) in AcOH (15 mL) at 80°C for 2 h gave the title compound (116 mg, 10 %) as an off-white solid after purification by flash column chromatography (x3) (elution with 0-40% MeOH/DCM gradient followed by 0-3% MeOH/DCM gradient and then 0-10% MeOH/DCM gradient ).

¹ H NMR (Methanol ₄ , 500 MHz) δ [ppm] = 8.48 (d, J=4.4 Hz, 1H), 7.95 (s, 1H), 7.79 (pseudo t, J=7.1 Hz, 1H), 7.48 (s, 2H), 7.39 (d, J=7.8 Hz, 1H), 7.36-7.26 (m, 1H), 7.25-7.12(m, 2H), 4.67(s, 2H), 3.89-3.78(m, 1H), 3.18-3.11(m, 2H), 3. 11-2.96(m, 5H), 2.73(q, J=8.6Hz, 1H), 2.49-2.39(m, 1H), 2.19-2.06(m, 1H).

HPLC-MS (Method C): [m/z]: 433.1 [M+H] ⁺ .

The above general scheme 9:

general technique 12:

tert-butyl 3-carbamoylazetidine-1-carboxylate (157) o

t-NBoc

1-[(tert-butoxy)carbonyl]azetidine-3-carboxylic acid (4.96 g, 24.65 mmol) and TEA (5.84 ml, 42.0 mmol) were dissolved in THF (60 ml) and cooled to -20°C. Isobutyl chloroformate (4.8 ml, 37.0 mmol) was added slowly and the reaction mixture was stirred <-10°C for 15 min. 28% aqueous ammonia solution (7.46 ml, 394 mmol) was added and the resulting mixture was warmed to room temperature and stirred for 1 h. The reaction mixture was quenched with saturated NaHCO ₃ solution and extracted with DCM (3x100 ml). The combined organic extracts were dried (Na ₂ SO ₄ ), filtered and evaporated in vacuo. Purification by flash column chromatography eluting with a gradient of 20-100% EtOAc/heptane followed by 1-4% MeOH/EtOAc gave the title compound (3.99 g, 81%) as a white solid.

¹ H NMR (CDCl3, 250 MHz): δ [ppm] = 5.47 (s, 2H), 4.22-4.01 (m, 4H), 3.34-3.15 (m, 1H), 1.46 (s, 9H).

HPLC-MS (method A): [m/z]: 222.95 [M+Na] ⁺ .

tert-Butyl 3-carbamoyl-3-methylazetidine-1-carboxylate (158)

Similar to general procedure 12, 1-[(tert-butoxy)carbonyl]-3-methylazetidine-3-carboxylic acid (110 mg, 0.51 mmol), TEA (90 mg, 0.87 mmol), isobutyl chloroformate (0. 1 ml, 0.77 mmol) and NH3 (28% aqueous solution, 0.15 ml, 10 mmol) in THF (10 ml) gave the title compound (102 mg, 93%) as a white solid.

¹ H NMR (CDCl3, 500 MHz): δ [ppm] = 5.63 (s, 1H), 5.37 (s, 1H), 4.21 (d, J=8.4 Hz, 2H ), 3.72 (d, J=8.4 Hz, 2H), 1.58 (s, 3H), 1.47 (s, 9H).

HPLC-MS (method A): [m/z]: 237 [M+Na] ⁺ .

tert-Butyl 3-carbamothioylazetidine-1-carboxylate (159)

- 149 039916

Similar to general procedure 11, Lawesson's reagent (4.43 g, 11.0 mmol), tert-butyl 3-carbamoyl azetidine-1-carboxylate (157) (3.99 g, 19.9 mmol) in DCM (60 ml) at room temperature within 30 min gave the title compound (4.47 g) as a pale yellow residue after flash chromatography through a pad of silica gel eluting with a 10-80% EtOAc/heptane gradient.

HPLC-MS (method A): [m/z]: 238.9 [M+Na]+.

tert-Butyl 3-carbamothioyl-3-methylazetidine-1-carboxylate (160)

Similar to general procedure 11, tert-butyl 3-carbamoyl-3-methylazetidine-1-carboxylate (158) (1.0 g, 4.67 mmol) and Lawsson's reagent (1.04 g, 3.0 mmol) in DCM ( 30 ml) gave the title compound (1.07 g) as a white solid.

1H NMR (CDCl ₃ , 250 MHz): δ [ppm] = 7.47 (s, 1H), 6.88 (s, 1H), 4.33 (d, J=8.3 Hz, 2H ), 3.81 (d, J=8.3 Hz, 2H), 1.72 (s, 3H), 1.47 (s, 9H).

HPLC-MS (Method A): [m/z]: 253.05 [M+Na]+.

Ethyl 2-{1-[(tert-butoxy)carbonyl]azetidin-3-yl}-1,3-thiazole-4-carboxylate (161) o

Eto -/ NBoc

L-S

Similar to general procedure 1, tert-butyl 3-carbamothioylazetidine-1-carboxylate (159) (4.47 g, 20.67 mmol), ethyl 3-bromo-2-oxopropanoate (2.85 ml, 22.73 mmol) and calcium carbonate (1.12 g, 11.0 mmol) in EtOH (40 mL) gave the title compound (3.54 g, 54%) as a yellow oil after purification by flash column chromatography eluting with a 10- 50% EtOAc/heptane.

1H NMR (DMSO-d6, 250 MHz): δ [ppm] = 8.49 (s, 1H), 4.37-4.23 (m, 5H), 4.00 (m, 2H), 1.41 (s, 9H), 1.31 (t, J=7.1 Hz, 3H).

HPLC-MS (method A): [m/z]: 335 [M+Na]+.

Ethyl 2-{1-[(tert-butoxy)carbonyl]-3-methylazetidin-3-yl}-1,3-thiazole-4-carboxylate (162) o

Eto Yao NBoc

Similar to general procedure 1, tert-butyl 3-carbamothioyl-3-methylazetidine-1-carboxylate (160) (0.82 g, 3.56 mmol), ethyl 3-bromo-2-oxopropanoate (0.49 ml, 4, 0 mmol) and calcium carbonate (0.2 g, 2.0 mmol) in EtOH (20 mL) gave the title compound (1.0 g) as a pale yellow oil.

1H NMR (CDCl ₃ , 500 MHz): δ [ppm] = 8.14 (s, 1H), 4.44 (q, J=7.1 Hz, 2H), 4.39 (d, J = 8.0 Hz, 2H), 3.98 (d, J=8.0 Hz, 2H), 1.83 (s, 3H), 1.48 (s, 9H), 1.43 (t, J =7.1 Hz, 3H).

HPLC-MS (method A): [m/z]: 349.15 [M+Na]+.

2-{1-[(tert-Butoxy)carbonyl]azetidin-3-yl}-1,3-thiazole-4-carboxylic acid (163)

Similar to general procedure 5, ethyl 2-{1-[(tert-butoxy)carbonyl]azetidin-3-yl}-1,3-thiazole-4carboxylate (161) (2.52 g, 7.91 mmol) and LiOH ( 0.57 g, 23.72 mmol) in THF (40 mL) and water (40 mL) gave the title compound (2.23 g, 87% pure, 86%) as an orange solid.

1H NMR (DMSO-H6, 500 MHz): δ [ppm] = 13.03 (s, 1H), 8.40 (s, 1H), 4.31-4.23 (m, 2H), 4.23-4.17(m, 1H), 4.05-3.93(m, 2H), 1.40(s, 9H).

HPLC-MS (Method A): [m/z]: 306.85 [M+Na]+.

2-{1-[(tert-Butoxy)carbonyl]-3-methylazetidin-3-yl}-1,3-thiazole-4-carboxylic acid (164) o ^HO Ypo NBoc

Similar to general procedure 5, ethyl 2-{1-[(tert-butoxy)carbonyl]-3-methylazetidin-3-yl}-1,3thiazol-4-carboxylate (162) (1.0 g, 3.06 mmol) and LiOH (0.22 g, 10.0 mmol) in THF (30 ml) and water (15 ml) gave the title compound (853 mg, 93%) as a pale yellow foam.

1H NMR (DMSO-d6, 250 MHz): δ [ppm] = 8.44 (s, 1H), 4.16 (d, J=8.2 Hz, 2H), 3.90 (d, J=8.2 Hz, 2H), 1.72(s, 3H), 1.41(s, 9H).

HPLC-MS (method A): [m/z]: 321.05 [M+Na]+.

tert-Butyl 3-(4-{[(3-fluoropyridin-2-yl)methyl]carbamoyl}-1,3-thiazol-2-yl)azetidine-1-carboxylate (165)

- 150 039916

Similar to general procedure 6, 2-{1-[(tert-butoxy)carbonyl]azetidin-3-yl}-1,3-thiazole-4carboxylic acid (163) (87%, 2.23 g, 6.69 mmol) , (3-fluoropyridin-2-yl)methanamine dihydrochloride (A2) (1.6 g, 8.03 mmol), DIPEA (4.66 ml, 26.77 mmol) and HATU (3.05 g, 8.03 mmol) in DCM (40 mL) gave the title compound (2.51 g, 82% pure, 79%) as an orange oil after purification by flash column chromatography eluting with a 20-100% EtOAc/heptane gradient.

1H NMR (DMSO-d6, 250 MHz): δ [ppm] = 8.79 (t, J=5.7 Hz, 1H), 8.38 (dt, J=4.6, 1.4 Hz, 1H), 8.24 (s, 1H), 7.76-7.62 (m, 1H), 7.46-7.35 (m, 1H), 4.66 (dd, J=5, 7, 1.5 Hz, 2H), 4.34-4.17 (m, 3H), 4.16-4.05 (m, 2H), 1.39 (s, 9H).

HPLC-MS (method A): [m/z]: 393 [M+H]+.

tert-Butyl 3-[4-({ 5H,6H,7H-cyclopenta[b]pyridin-7-yl}carbamoyl)-1,3-thiazol-2-yl]azetidine-1 carboxylate (166) /gHH.H___ .N /x // / ΰ Η X—< NBoc \^N ^{H 47}

Similar to general procedure 6, 2-{1-[(tert-butoxy)carbonyl]azetidin-3-yl}-1,3-thiazole-4carboxylic acid (163) (300mg, 1.06mmol), 5H,6H, 7H-cyclopenta[b]pyridine-7-amine dihydrochloride (284 mg, 1.37 mmol), DIPEA (0.61 ml, 3.5 mmol) and HATU (0.52 g, 1.4 mmol) in DCM ( 20 ml) gave the crude title compound (1.32 g, 30% pure, 94%) which was used in the next step without purification.

HPLC-MS (method A): [m/z]: 401.1 [M+H] ⁺ .

tert-Butyl 3-(4-{[(3-chloropyridin-2-yl)methyl]carbamoyl}-1,3-thiazol-2-yl)azetidine-1-carboxylate (167)

CI o ί U N

Similar to general procedure 6, 2-{1-[(tert-butoxy)carbonyl]azetidin-3-yl}-1,3-thiazole-4carboxylic acid (163) (364 mg, 1.28 mmol), (3-chloropyridine -2-yl)methanamine dihydrochloride (359 mg, 1.66 mmol), DIPEA (0.74 ml, 4.2 mmol) and HATU (0.63 g, 1.7 mmol) in DCM (20 ml) gave the indicated title compound (493 mg, 94%) as a pale yellow oil after purification by flash column chromatography (eluting with a 0-100% EtOAc/heptane gradient).

1H NMR (DMSO-d ₆ , 250 MHz): δ [ppm] = 8.79 (t, J=5.5 Hz, 1H), 8.51 (dd, J=4.7, 1, 4 Hz, 1H), 8.27 (s, 1H), 7.96 (dd, J=8.1, 1.4 Hz, 1H), 7.39 (dd, J=8.1, 4.7 Hz, 1H), 4.70 (d, J=5.5 Hz, 2H), 4.38-4.18 (m, 3H), 4.09 (q, J=3.9 Hz, 2H), 1.41 (s, 9H).

HPLC-MS (method A): [m/z]: 409.05 [M+H] ⁺ .

tert-Butyl 3-(4-{[(3-fluoropyridin-2-yl)methyl]carbamoyl}-1,3-thiazol-2-yl)-3-methylazetidine-1 carboxylate(168)

F O

Similar to general procedure 6, 2-{1-[(tert-butoxy)carbonyl]-3-methylazetidin-3-yl}-1,3-thiazole-4-carboxylic acid (164) (546 mg, 1.83 mmol), ( 3-fluoropyridin-2-yl)methanamine dihydrochloride (A2) (400 mg, 2.01 mmol), DIPEA (1.05 ml, 6.0 mmol) and HATU (830 mg, 2.0 mmol) in DCM (20 ml) gave the crude title compound (824 mg) as a pale yellow oil. The resulting crude product was used in the next step without further purification.

1H NMR (CDCl ₃ , 500 MHz): δ [ppm] = 8.38-8.33 (m, 1H), 8.28 (s, 1H), 8.02 (s, 1H), 7 .37-7.31 (m, 1H), 7.23-7.17 (m, 1H), 4.81-4.76 (m, 2H), 4.30 (d, J=7.5 Hz , 2H), 3.88 (d, J=7.5 Hz, 2H), 1.52 (s, 3H), 1.40 (s, 9H).

HPLC-MS (method A): [m/z]: 407.2 [M+H] ⁺ .

2-(Azetidin-3-yl)-N-[(3-fluoropyridin-2-yl)methyl]-1,3-thiazol-4-carboxamide dihydrochloride (169)

Similar to general procedure 4, tert-butyl 3-(4-{[(3-fluoropyridin-2-yl)methyl]carbamoyl}-1,3thiazol-2-yl)azetidine-1-carboxylate (165) (82%, 2 .51 g, 5.26 mmol) and 12 M HC (8.8 mL) in MeOH (25 mL) gave the title compound (2.26 g, 85% pure, quant) as a light brown solid .

1H NMR (Methanol^, 250 MHz): δ [ppm] = 8.56 (d, J=5.2 Hz, 1H), 8.27 (s, 1H), 8.24-8.07 (m, 1H), 7.87-151 039916

7.73 (m, 1H), 4.96-4.91 (m, 2H), 4.57-4.41 (m, 5H).

HPLC-MS (Method A): [m/z]: 292.95 [M+H]+.

2-(Azetidin-3-yl)-N-{5Н,6Н,7Н-cyclopenta[b]pyridin-7-yl}-1,3-thiazol-4-carboxamide (170)

As in general procedure 4, crude tert-butyl 3-[4-({5H,6H,7H-cyclopenta[b]pyridin-7yl}carbamoyl)-1,3-thiazol-2-yl]azetidine-1-carboxylate (166 ) (1.32 g, 30% pure, 1.06 mmol) and 12M HC (1 ml) in MeOH (10 ml) gave the title compound (322 mg, quant) as a white solid after purification on SCX -2 cartridge, washing with DCM and MeOH, then eluting with 7N. a solution of ammonia in MeOH.

HPLC-MS (method A): [m/z]: 300.95 [M+H] ⁺ .

2-(Azetidin-3-yl)-N-[(3-chloropyridin-2-yl)methyl]-1,3-thiazol-4-carboxamide (171)

Similar to general procedure 4, tert-butyl 3-(4-{[(3-chloropyridin-2-yl)methyl]carbamoyl}-1,3thiazol-2-yl)azetidine-1-carboxylate (167) (423 mg, 1 .03 mmol) and 12M HC (2 ml) in MeOH (30 ml) gave the title compound (330 mg, 99%) as a white solid after purification on an SCX2 cartridge, washing with DCM and MeOH, then eluting with 7N. a solution of ammonia in MeOH.

¹ H NMR (DMSO-d ₆ , 500 MHz): δ [ppm] = 8.71 (t, J=5.5 Hz, 1H), 8.52 (dd, J=4.7, 1 .4 Hz, 1H), 8.24 (s, 1H), 7.96 (dd, J=8.0, 1.4 Hz, 1H), 7.39 (dd, J=8.0, 4, 7 Hz, 1H), 4.70 (d, J=5.5 Hz, 2H), 4.24 (m, 1H), 3.92-3.87 (m, 2H), 3.76-3, 73 (m, 2H).

HPLC-MS (method A): [m/z]: 308.95 [M+H] ⁺ .

N-[(3-Fluoropyridin-2-yl)methyl]-2-(3-methylazetidin-3-yl)-1,3-thiazol-4-carboxamide (172)

Similar to general procedure 4, tert-butyl 3-(4-{[(3-fluoropyridin-2-yl)methyl]carbamoyl}-1,3thiazol-2-yl)-3-methylazetidine-1-carboxylate (168) (824 mg, 2.03 mmol) and 12M HC (2 ml) in MeOH (20 ml) gave the title compound (476 mg, 74%) after purification on an SCX-2 cartridge, washing with DCM and MeOH, then eluting with 7N. a solution of ammonia in MeOH.

1H NMR (DMSO-66, 250 MHz): δ [ppm] = 8.71 (t, J=5.7 Hz, 1H), 8.39 (dt, J=4.6, 1.3 Hz, 1H), 8.21 (s, 1H), 7.71 (ddd, J=10.0, 8.3, 1.3 Hz, 1H), 7.46-7.36 (m, 1H) , 4.68 (m, 2H), 3.85 (d, J=7.5 Hz, 2H), 3.52 (d, J=7.5 Hz, 2H), 1.72 (s, 3H) .

HPLC-MS (method A): [m/z]: 307.20 [M+H] ⁺ .

2-{1-[2-(1H-1,3-Benzodiazol-2-yl)ethyl]azetidin-3-yl}-N-[(3-fluoropyridin-2-yl)methyl]-1,3-thiazole4 -carboxamide (illustrative example No. 193)

Similar to general procedure 8, 2-(azetidin-3-yl)-X-[(3-fluoropyridin-2-yl)methyl]-1,3-thiazole-4-carboxamide dihydrochloride (169) (85%, 2.26 g, 5.26 mmol), N-(2-nitrophenyl)prop-2-enamide (D) (1.21 g, 6.31 mmol) and DBU (2.35 ml, 15.77 mmol) in MeCN (30 ml ) gave a crude intermediate which was reacted with iron powder (0.51 g, 9.0 mmol) and AcOH (5 ml) to give the title compound (122 mg, 7%) as a cream-colored solid after purification by method basic prepHPLC.

1H NMR (Methanol-d ₄ , 500 MHz): δ [ppm] = 8.33 (dt, J=4.7, 1.3 Hz, 1H), 8.12 (s, 1H), 7 .62-7.56 (m, 1H), 7.53-7.47 (m, 2H), 7.39-7.34 (m, 1H), 7.22-7.17 (m, 2H) , 4.79 (d, J=1.7 Hz, 2H), 4.11-4.03 (m, 1H), 3.80 (t, J=7.9 Hz, 2H), 3.55- 3.50 (m, 2H), 3.07-3.02 (m, 2H), 3.01-2.96 (m, 2H).

HPLC-MS (method d): [m/z]: 437 [M+H] ⁺ .

2-{1-[2-(1H-1,3-Benzodiazol-2-yl)ethyl]azetidin-3-yl}-N-{5H,6H,7H-cyclopenta[b]pyridin-7-yl}1 ,3-thiazole-4-carboxamide (illustrative example No. 194)

Similar to general procedure 8, 2-(azetidin-3-yl)-N-{5H,6H,7H-cyclopenta[b]pyridin-7-yl}-1,3thiazol-4-carboxamide (170) (322 mg, 1 .07 mmol), N-(2-nitrophenyl) prop-2-enamide (D) (227 mg, 1.18

- 152039916 mmol) and DBU (0.21 ml, 1.4 mmol) in MeCN (10 ml) gave a crude intermediate which was reacted with iron powder (0.15 g, 2.6 mmol) in AcOH (4 ml ) to give the title compound (139 mg, 48%) as a white solid after purification by basic prep-HPLC.

1H NMR (Methanol-b ₄ , 500 MHz): δ [ppm] = 8.35 (d, J=5.0 Hz, 1H), 8.15 (s, 1H), 7.78-7 .74 (m, 1H), 7.50 (brs, 2H), 7.29 (dd, J=7.6, 5.0 Hz, 1H), 7.24-7.18 (m, 2H ), 5.58 (t, J=8.3 Hz, 1H), 4.05 (t, J=7.9, 6.4 Hz, 1H), 3.77 (t, J=7.9 Hz , 2H), 3.56-3.49 (m, 2H), 3.11 (ddd, J=16.3, 9.1, 2.8 Hz, 1H), 3.05-3.00 (m , 3H), 3.00-2.94 (m, 2H), 2.73 (m, 1H), 2.11 (m, 1H).

HPLC-MS (method c): [m/z]: 445.3 [M+H] ⁺ .

2-{1-[2-(1H-1,3-Benzodiazol-2-yl)ethyl]azetidin-3-yl}-N-[(3-chloro-pyridin-2-yl)methyl]-1,3-thiazole4 -carboxamide (illustrative example No. 195)

Similar to general procedure 8, 2-(azetidin-3-yl)-H-[(3-chloropyridin-2-yl)methyl]-1,3-thiazole-4-carboxamide (171) (330 mg, 1.07 mmol), Na(2-nitrophenyl)prop-2-enamide (D) (226 mg, 1.18 mmol) and DBU (0.21 ml, 1.4 mmol) in MeCN (10 ml) gave the desired crude intermediate which reacted with iron powder (150 mg, 2.6 mmol) in AcOH (4 mL) to give the title compound (80 mg, 27%) as a white solid after purification by basic prep-HPLC, followed by flash column chromatography (elution with a 0-12% MeOH/DCM gradient).

¹ H NMR (Methanol-b ₄ , 500 MHz): δ [ppm] = 8.45 (dd, J=4.7, 1.4 Hz, 1H), 8.15 (s, 1H), 7.88 (dd, J=8.1, 1.4 Hz, 1H), 7.51 (broad s, 2H), 7.32 (dd, J=8.1, 4.7 Hz, 1H) , 7.24-7.19 (m, 2H), 4.85-4.82 (m, 2H), 4.11 (m, 1H), 3.84 (t, J=7.9 Hz, 2H ), 3.59-3.53 (m, 2H), 3.10-3.05 (m, 2H), 3.03-2.98 (m, 2H).

HPLC-MS (Method C): [m/z]: 453.2 [M+H]+.

2-{1-[2-(1H-1,3-Benzodiazol-2-yl)ethyl]-3-methylazetidin-3-yl}-N-[(3-fluoropyridin-2-yl)methyl] 1.3 -thiazole-4-carboxamide (illustrative example No. 212)

Similar to general procedure 8, N-[(3-fluoropyridin-2-yl)methyl]-2-(3-methylazetidin-3-yl)-1,3thiazol-4-carboxamide (172) (476 mg, 1.55 mmol ), N-(2-nitrophenyl)prop-2-enamide (D) (328 mg, 1.71 mmol) and DBU (0.26 mL, 2 mmol) in MeCN (15 mL) gave the desired crude intermediate, which reacted with iron powder (350 mg, 10.0 mmol) in AcOH (5 ml). Purification by basic prep-HPLC followed by flash column chromatography (eluting with a 0-30% MeOH/DCM gradient) gave the title compound (162 mg, 23%) as a white solid.

1H NMR (Methanol-04, 500 MHz): δ [ppm] = 8.34-8.27 (m, 1H), 8.14 (s, 1H), 7.60 (ddd, J=9 .7, 8.5, 1.2 Hz, 1H), 7.51 (s, 2H), 7.37-7.32 (m, 1H), 7.24-7.18 (m, 2H), 4.80 (d, J=1.3 Hz, 2H), 3.78 (d, J=7.7 Hz, 2H), 3.47 (d, J=7.7 Hz, 2H), 3. 10 (t, J=7.2 Hz, 2H), 3.00 (t, J=7.2 Hz, 2H), 1.78 (s, 3H).

HPLC-MS (method C): [m/z]: 451.1 [M+H] ⁺ .

The above general scheme 10:

methyl 2,2-dimethyl-3-oxopropanoate (173)

In ice-cold (0°C) solution of methyl 3-hydroxy-2,2-dimethylpropanoate (1.45 ml, 11.35 mmol) and TEA (4.75 ml, 34.05 mmol) in DCM (45 ml) and DMSO (8 ml, 113.5 mmol) was added a complex of pyridine with sulfur trioxide (5.42 g, 34.05 mmol). The resulting mixture was stirred at room temperature for 22 hours. The reaction was quenched with a saturated solution of NH ₄ Cl (30 ml). The layers were separated and the aqueous layer was extracted with dichloromethane (40 ml). The organic layer was washed successively with 2M HC solution (2x20 ml) and saturated sodium chloride solution (15 ml), dried (MgSO ₄ ), filtered and evaporated to give the title compound (0.35 g, 99%) as an orange oil.

1H NMR (CDCl ₃ , 250 MHz): δ [ppm] = 9.66 (s, 1H), 5.30 (s, 1H), 3.75 (s, 3H), 1.35 (s , 6H).

Methyl

3-{[2-(4-{[(3-Fluoropyridin-2-yl)methyl]carbamoyl}-1,3-thiazol-2-yl)ethyl]amino}-2,2 dimethylpropanoate (174)

DIPEA (1.8 ml, 10.32 mmol) was added to a solution of 2-(2-aminoethyl)-N-[(3-fluoropyridin-2-yl)methyl]- 153 039916

1,3-thiazole-4-carboxamide dihydrochloride (103) (0.97 g, 2.58 mmol) and methyl 2,2-dimethyl-3-oxopropanoate (173) (0.52 g, 3.36 mmol) in MeOH ( 20 ml). The reaction mixture was stirred at room temperature for 18 hours. The reaction mixture was cooled to 0° C. and NaBH ₄ (146 mg, 3.87 mmol) was added portionwise. The reaction mixture was allowed to warm to room temperature and stirred for 3.5 h. The solvent was evaporated and water (10 ml) was added. The aqueous layer was extracted with ethyl acetate (3x20 ml). The combined organic layers were dried (MgSO ₄ ), filtered and evaporated to give an orange oil (1.1 g). Purification by flash column chromatography (eluting with a 0-15% MeOH/DCM gradient) gave the title compound (0.95 g, 79%) as a pale yellow oil.

1H NMR (CDCl ₃ , 250 MHz): δ [ppm] = 8.43-8.38 (m, 2H), 8.02 (s, 1H), 7.45-7.36 (m, 1H), 7.26-7.20 (m, 1H), 4.85 (dd, J=5.3, 1.7 Hz, 2H), 3.63 (s, 3H), 3.20 (t , J=5.6 Hz, 2H), 3.10 (t, J=5.8 Hz, 2H), 2.79 (s, 2H), 1.22 (s, 6H).

HPLC-MS (method A): [m/z]: 395.05 [M+H]+.

3-{[(tert-Butoxy)carbonyl][2-(4-{[(3-fluoropyridin-2-yl)methyl]carbamoyl}-1,3-thiazol-2yl)ethyl]amino} -2,2- dimethylpropanoate (175)

TEA (0.5 ml, 3.62 mmol) was added to methyl 3-{[2-(4-{[(3-fluoropyridin-2-yl)methyl]carbamoyl}-1,3thiazol-2-yl)ethyl] amino}-2,2-dimethylpropanoate (174) (0.95 g, 2.41 mmol) in DCM (10 mL). The reaction mixture was stirred for 5 min at room temperature, di-tert-butyl dicarbonate (0.63 g, 2.9 mmol) in DCM (10 ml) was added dropwise and the reaction mixture was stirred at room temperature for 20 h. The solvent was evaporated and purified by flash column chromatography (eluting with a 0-100% EtOAc/heptane gradient) to give the title compound (0.94 g, 90%) as a clear oil.

¹ H NMR (CDCl ₃ , 500 MHz): δ [ppm] = 8.42 (d, J=4.3 Hz, 2H), 8.01 (s, 1H), 7.44-7, 37 (m, 1H), 7.26 (m, 1H, overlap with solvent signals), 4.85 (dd, J=5.1, 1.7 Hz, 2H), 3.70 (s, 3H), 3.55 (t, J=7.0 Hz, 2H), 3.48-3.41 (breadth m, 2H), 3.31-3.14 (m, 2H), 1.45 (s, 9H), 1.19 (s, 6H).

HPLC-MS (method A): [m/z]: 495.15 [M+H] ⁺ .

3-{[(tert-Butoxy)carbonyl][2-(4-{[(3-fluoropyridin-2-yl)methyl]carbamoyl}-1,3-thiazol-2yl)ethyl]amino}-2,2- dimethylpropanoic acid (176) f o

[|4^nU< ^N ^_ %-OtBu

Ui u ^- he oh

Similarly, using General Procedure 5, LiOH (0.27 g, 11.1 mmol) and 3-{[(tert-butoxy)carbonyl][2-(4-{[(3-fluoropyridin-2-yl)methyl]carbamoyl }-1,3-thiazol-2-yl)ethyl]amino}-2,2dimethylpropanoate (175) (0.94 g, 1.85 mmol) in THF (6 ml)/water (1.5 ml) gave the title compound (0.91 g, 96%) as an off-white solid.

¹ H NMR (DMSO-dg, 500 MHz): δ [ppm] = 12.41 (s, 1H), 8.63 (s, 1H), 8.39 (s, 1H), 8.17 (s, 1H), 7.71 (ddd, J=10.0, 8.3, 1.2 Hz, 1H), 7.41 (dt, J=8.6, 4.4 Hz, 1H), 4.67 (dd, J=5.6, 1.3 Hz, 2H), 3.53 (t, J=7.1 Hz, 2H), 3.38 (s, 2H), 3.23 (t , J=7.1 Hz, 2H), 1.34 (s, 9H), 1.07 (s, 6H).

HPLC-MS (method A): [m/z]: 481.15 [M+H] ⁺ .

tert-Butyl N-{2-[(2-aminophenyl)carbamoyl]-2,2-dimethylethyl}-N-[2-(4-{[(3-fluoropyridin-2yl)methyl]carbamoyl} -1,3- thiazol-2-yl)ethyl] carbamate (177)

Similarly, using the general procedure 6, 3-{[(tert-butoxy)carbonyl] [2-(4-{[(3fluoropyridin-2-yl)methyl]carbamoyl}-1,3-thiazol-2-yl)ethyl ]amino}-2,2-dimethylpropanoic acid (176) (0.5 g, 0.98 mmol), benzene-1,2-diamine (0.14 g, 1.3 mmol), TEA (0.18 ml , 1.3 mmol) and HATU (0.49 g, 1.3 mmol) in DMF (10 ml) gave the title compound (0.51 g, 83%) as a yellow oil after purification by flash column chromatography using a 0-100% EtOAc/heptane gradient.

1H NMR (DMSO-d ₆ , 250 MHz): δ [ppm] = 8.84 (s, 1H), 8.61 (s, 1H), 8.37 (d, J=4.5 Hz , 1H), 8.16 (s, 1H), 7.70 (ddd, J=10.1, 8.3, 1.2 Hz, 1H), 7.41 (dt, J=8.6, 4 .4 Hz, 1H), 7.02-6.86 (m, 2H), 6.78-6.68 (m, 1H), 6.57-6.45

- 154 039916 (m, 1H), 4.66 (d, J=4.4 Hz, 2H), 3.55 (t, J=6.9 Hz, 2H), 3.50 (s, 2H), 3.24 (t, J=6.9 Hz, 2H), 1.35 (s, 9H), 1.21 (s, 6H).

HPLC-MS (method A): [m/z]: 571.2 [M+H]+.

tert-Butyl N[2-(1H-1,3-benzodiazol-2-yl)-2-methylpropyl]-N-[2-(4-{[(3-fluoropyridin-2yl)methyl]carbamoyl}-1, 3-thiazol-2-yl)ethyl]carbamate (178)

tert-Butyl N-{2-[(2-aminophenyl)carbamoyl]-2,2-dimethylethyl}-N-[2-(4-{[(3-fluoropyridin-2yl)methyl]carbamoyl}-1,3- thiazol-2-yl)ethyl]carbamate (177) (260 mg, 0.415 mmol) in ice-cold AcOH (4 ml) was heated at 80°C for 35 min. The reaction mixture was cooled, evaporated in vacuo and the resulting residue was partitioned between EtOAc (10 ml) and saturated K2CO3 solution (10 ml). The layers were separated and the aqueous layer was extracted with ethyl acetate (2x10 ml). The combined organic layers were dried (Na ₂ SO ₄ ), filtered and evaporated to give a brown oil. Purification by flash column chromatography (KP-NH column, eluting with a 0-100% DCM/heptane gradient) gave the title compound (172 mg, 71%) as a pale brown oil.

1H NMR (CDCl3, 500 MHz): δ [ppm] = 8.43 (brs, 1H), 8.42 (dt, J=4.6 and 1.2 Hz, 2H), 7, 99 (s, 1H), 7.82-7.69 (m, 1H), 7.47-7.42 (m, 1H), 7.40-7.34 (m, 1H), 7.25- 7.18 (m, 2H), 4.91 (d, J=4.5 Hz, 2H), 3.75-3.69 (br. m, 2H), 3.59-3.39 (b.w. m, 2H), 3.11 (t, J=6.3 Hz, 2H), 1.59 (s, 9H), 1.45 (s, 6H).

HPLC-MS (method A): [m/z]: 553.2 [M+H] ⁺ .

2-(2-{[2-(1H-1,3-Benzodiazol-2-yl)-2-methylpropyl]amino}ethyl)-N-[(3-fluoropyridin-2-yl)methyl]1,3- thiazole-4-carboxamide (illustrative example No. 199)

Similar to general procedure 2, TFA (2 ml) was added to a solution of tert-butyl N-{2-[(2aminophenyl)carbamoyl]-2,2-dimethylethyl}-N-[2-(4-{[(3-fluoropyridine- 2-yl)methyl]carbamoyl}-1,3thiazol-2-yl)ethyl]carbamate (178) (172 mg, 0.25 mmol) in DCM (2 ml) and stirred at room temperature for 3 hours to give the title title compound (79 mg, 70%) as a beige solid after purification on an Isolute SCX-2 cartridge, eluting with DCM (2 column volumes), DCM/MeOH (1:1, 1 column volume), MeOH (2 column volumes column) and then 7M NH3 in MeOH (2 column volumes).

1H NMR (DMSO-d ₆ , 500 MHz): δ [ppm] = 12.01 (s, 1H), 8.62 (t, J=5.7 Hz, 1H), 8.37 (dt , J=4.7 and 1.5 Hz, 1H), 8.02 (s, 1H), 7.74-7.63 (m, 1H), 7.53-7.35 (m, 2H), 7.42-7.38 (m, 1H), 7.14-7.06 (m, 2H), 4.64 (dd, J=5.6, 1.4 Hz, 2H), 3.11 (t, J=6.5 Hz, 2H), 2.91-2.87 (m, 4H), 1.39 (s, 6H).

HPLC-MS (Method D): [m/z]: 453.2 [M+H] ⁺ .

2-(1H-1,3-Benzodiazol-2-yl)acetate (179) rfe

2M NaOH (90.7 ml, 181 mmol) was added to 2-(1H-1,3-benzodiazol-2-yl)acetonitrile (9.5 g, 60.4 mmol). The reaction mixture was heated at 100° C. for 3 hours. The reaction mixture was cooled to room temperature and acidified to pH 5-6 with 4M HC. The precipitate that formed was filtered off, washed with water and dried in vacuo to give the title compound (9.7 g, 77%) as a light brown solid.

1H NMR (DMSO-d ₆ , 500 MHz): δ [ppm] = 7.52-7.46 (m, 1H), 7.45-7.39 (m, 1H), 7.17- 7.11 (m, 1H), 7.117.06 (m, 1H), 2.47 (s, 2H).

HPLC-MS (method F): [m/z]: 177.0 [M+H] ⁺ .

Methyl 2-(1H-1,3-benzodiazol-2-yl)acetate (180)

Thionyl dichloride (0.7 ml, 9.65 mmol) was added dropwise to an ice-cold (0°C) suspension of 2-(1H-1,3-benzodiazol-2-yl)acetic acid (179) (0.7 g, 3. 97 mmol) in MeOH (30 ml). The reaction mixture was allowed to warm to room temperature and stirred for 18 hours. The resulting mixture was poured into saturated NaHCO ₃ solution (40 ml) and extracted with DCM (3x20 ml). The combined organic layers were washed with saturated sodium chloride solution (30 ml), dried (NaSO ₄ ), filtered and evaporated to give the title compound (0.65 g, 86%) as an off-white solid.

1H NMR (CDCl ₃ , 500 MHz): δ [ppm] = 10.10 (br s, 1H), 7.72 (br s, 1H), 7.46 (br s, 1H) , 7.297.23 (m, 2H), 4.09 (s, 2H), 3.82 (s, 3H).

HPLC-MS (method F): [m/z]: 191.2 [M+H] ⁺ .

tert-Butyl 2-(2-methoxy-2-oxoethyl)-1Н-1,3-benzodiazol-1-carboxylate (181)

Similar to general procedure 4, methyl 2-(1H-1,3-benzodiazol-2-yl)acetate (180) (650 mg, 3.42 mmol), TEA (0.5 ml, 3.59 mmol), Boc ₂ O (890 mg, 4.11 mmol) and DMAP (84 mg, 0.68 mmol) in THF (30 mL) at room temperature over 18 h gave the title compound (940 mg, 95%) as a solid cream color after purification by flash column chromatography (elution with a gradient of 0-100% EtOAc/heptane).

1H NMR (CDCl ₃ , 500 MHz): δ [ppm] = 7.96-7.90 (m, 1H), 7.74-7.68 (m, 1H), 7.38-7, 31 (m,

2H), 4.28 (s, 2H), 3.73 (s, 3H), 1.69 (s, 9H).

HPLC-MS (method A): [m/z]: 291 [M+H]+.

tert-Butyl 2-(1,1-difluoro-2-methoxy-2-oxoethyl)-1 H-1,3-benzodiazol-1-carboxylate (182)

KHMDS (1.5 g, 7.61 mmol) in THF (15 ml) was cooled to -78°C. tert-Butyl 2-(2-methoxy2-oxoethyl)-1H-1,3-benzodiazol-1-carboxylate (181) (0.74 g, 2.54 mmol) was added and the resulting mixture was stirred at -78°C for within 45 min. A mixture of 18-crown-6 (2.01 g, 7.61 mmol) and Ν-φτορ-Ν(phenylsulfonyl)benzenesulfonamide (2.2 g, 7.1 mmol) was added. The resulting mixture was stirred at -78°C for 30 minutes. More KHMDS (0.75 g, 3.76 mmol) was added and stirred for 20 minutes. More N-fluoro-N-phenylsulfonyl)benzenesulfonamide (1.1 g, 3.49 mmol) and 18-crown-6 (1 g, 3.78 mmol) were added. The reaction mixture was then allowed to slowly warm to room temperature and stirred for 18 hours to give a 1.6:1 ratio of bis:monofluorinated products. Saturated NH ₄ Cl solution (12 ml) was added followed by water (20 ml). The organic and aqueous layers were separated. The aqueous layer was extracted with EtOAc (3x50 ml). The combined organic layers were washed with saturated sodium chloride solution (100 ml), dried (NaSO ₄ ), filtered and evaporated to give a pink semi-solid (3 g). Purification by flash column chromatography (eluting with a gradient of 0-50% TBME/heptane, then 100% TBME) gave the title compound (0.47 g, 48%) as a yellow oil which solidified on standing.

1H NMR (CDCl ₃ , 250 MHz): δ [ppm] = 8.00-7.81 (m, 2H), 7.55-7.34 (m, 2H), 3.91 (s, 3H), 1.70 (s, 9H). HPLC-MS (method A): [m/z]: 226.9 [M-BOC+H] ⁺ .

tert-Butyl 2-(1,1-difluoro-2-hydroxy-2-methoxyethyl)-1H-1,3-benzodiazol-1-carboxylate (183)

NaBH ₄ (44 mg, 1.16 mmol) was added to an ice-cold (0°C) solution of tert-butyl 2-(1,1-difluoro-2-methoxy2-oxoethyl)-1H-1,3-benzodiazol-1-carboxylate (182) (295 mg, 0.76 mmol) in EtOH (3.5 ml). The reaction mixture was stirred at 0°C for 1.5 h, quenched by dropping 1 M HC (0.5 ml). Water (10 ml) was added and the aqueous layer was extracted with ethyl acetate (3x10 ml). The combined organic layers were dried (NaSO ₄ ), filtered and evaporated to give a white solid (300 mg). Purification by flash column chromatography (eluting with a 0-40% EtOAc/heptane gradient) gave the title compound (185 mg, 59%) as a white solid.

1H NMR (CDCl ₃ , 250 MHz): δ [ppm] = 8.03-7.77 (m, 1H), 7.63-7.31 (m, 3H), 6.17 (dd, J=6.4, 4.9 Hz, 1H), 3.69 (s, 3H), 1.52 (s, 9H).

- 156 039916

HPLC-MS (method A): [m/z]: 329.05 [M+H]+.

4-(2-{[2-(1H-1,3-Benzodiazol-2-yl)-2,2-difluoroethyl]amino}ethyl)-N-[(3-fluoropyridin-2-yl)methyl] 1, 3-thiazole-2-carboxamide (illustrative example No. 196)

tert-Butyl 2-(1,1-difluoro-2-hydroxy-2-methoxyethyl)-1H-1,3-benzodiazol-1-carboxylate (183) (160 mg, 0.4 mmol), 2-(2- aminoethyl)-N-[(3-fluoropyridin-2-yl)methyl]-1,3-thiazole-4-carboxamide (103) (136 mg, 0.49 mmol) and activated molecular sieves in dry toluene (3 ml) heated at 125° C. for 18 hours. The reaction mixture was cooled to room temperature and NaBH4 (40 mg, 1.06 mmol) was added followed by EtOH (4 ml). The reaction mixture was stirred for 1 hour. The reaction was quenched by dropping 1 M HC (10 drops). At the end of gas evolution, water (10 ml) was added and the pH was adjusted to 10 with 2M K ₂ CO ₃ solution. The resulting mixture was filtered and the aqueous layer was extracted with ethyl acetate (3x20 ml). The combined organic layers were dried (NaSO ₄ ), filtered and evaporated to give a brown oily solid (176 mg). Purification by flash column chromatography (eluting with a 0-10% MeOH/DCM gradient) gave a brown solid (40 mg). Further purification by basic prep-HPLC gave a yellow oil (21 mg). Purification by flash column chromatography (eluting with a 0-5% EtOH/DCM gradient) gave the title compound (12 mg, 6%) as a pale yellow solid.

1H NMR (Methanol-d ₄ , 250 MHz): δ [ppm] = 8.38-8.32 (m, 1H), 7.90 (s, 1H), 7.65-7.55 ( m, 3H), 7.42-7.36 (m, 1H), 7.36-7.30 (m, 2H), 4.73 (d, J=1.6 Hz, 2H), 3.60 (t, J=14.0 Hz, 2H), 3.20-3.08 (m, 4H).

HPLC-MS (Method D): [m/z]: 461.2 [M+H] ⁺ .

The above general scheme 11:

general technique 12:

ethyl 2-(2-{[(tert-butoxy)carbonyl]amino}ethyl)-5-chloro-1,3-thiazole-4-carboxylate (184)

A solution of ethyl 2-(2-{[(tert-butoxy)carbonyl]amino}ethyl)-1,3-thiazole-4-carboxylate (1) (2 g, 6.66 mmol) and hexachloroethane (1.49 g, 6.32 mmol) in THF (140 ml) was cooled to -78°C. A 2M NaHMDS solution (7.0 ml, 14.0 mmol) was added dropwise. The reaction mixture was stirred at -78°C for 1 h, quenched with saturated aqueous NH ₄ Cl and left to warm to room temperature. The resulting mixture was filtered and the resulting residue was washed with THF (50 ml). The filtrates were evaporated in vacuo and the resulting residue was partitioned between EtOAc and saturated aqueous NH ₄ Cl. The organic layer was separated, dried (MgSO ₄ ) and evaporated in vacuo. Purification by flash chromatography (eluting with a 0-30% EtOAc/DCM gradient) gave the title compound (1.47 g, 65%) as a white solid.

1H NMR (CDCl ₃ , 250 MHz): δ [ppm] = 4.91 (s, 1H), 4.45 (q, 7 = 7.1 Hz, 2H), 3.55 (q, J =6.3 Hz, 2H), 3.18 (t, J=6.4 Hz, 2H), 1.49-1.39 (m, 12H).

HPLC-MS (Method A): [m/z]: 356.95 [M+H] ⁺ .

2-(2-{[(tert-Butoxy)carbonyl]amino}ethyl)-5-chloro-1,3-thiazole-4-carboxylic acid (185)

Similarly, using General Procedure 5, LiOH (739 mg, 30.84 mmol) and ethyl 2-(2{[(tert-butoxy)carbonyl]amino}ethyl)-5-chloro-1,3-thiazole-4- carboxylate (184) (1.47 g, 4.41 mmol) in THF (30 ml)/water (10 ml) gave the title compound (1.35 g, quant.) as a yellow oil. The resulting compound was used in the next step without further purification.

1H NMR (CDCl3, 500 MHz): δ [ppm] = 4.92 (s, 1H), 3.58 (q, J=5.7 Hz, 2H), 3.18 (t, J= 5.8 Hz, 2H), 1.46 (s, 9H).

HPLC-MS (method A): [m/z]: 328.9 [M+H] ⁺ .

tert-Butyl N-[2-(5-chloro-4-{[(3-fluoropyridin-2-yl)methyl]carbamoyl}-1,3-thiazol-2-yl)ethyl]carbamate (186)

Similarly, using the general procedure 6, 2-(2-{[(tert-Butoxy)carbonyl]amino}ethyl)-5-157 039916 chloro-1,3-thiazole-4-carboxylic acid (185) (800 mg, 2.61 mmol), (3-fluoropyridin-2-yl)methanamine dihydrochloride (A2) (623 mg, 3.13 mmol), DIPEA (1.5 ml, 10.0 mmol), HATU (1.19 g, 3.0 mmol) in DCM (30 mL) gave the title compound (1.08 g, 80%) as a white solid after purification by flash column chromatography (eluting with a 0-60% EtOAc/heptane gradient). The compound was used in the next step without further purification.

1H NMR (CDCl3, 250 MHz): δ [ppm] = 8.49-8.39 (m, 2H), 7.44 (ddd, J=9.4, 8.4, 1.3 Hz , 1H), 7.34-7.23 (m, 3H), 4.94 (s, 1H), 4.84 (dd, J=5.1, 1.7 Hz, 2H), 3.59 ( q, J=6.3 Hz, 2H), 3.15 (t, J=6.4 Hz, 2H), 1.46 (s, 9H).

HPLC-MS (method A): [m/z]: 415 [M+H] ⁺ .

tert-Butyl N-[2-(5-chloro-4-{[(3-chloropyridin-2-yl)methyl]carbamoyl}-1,3-thiazol-2-yl)ethyl]carbamate (187)

Similarly, using the general procedure 6, 2-(2-{[(tert-butoxy)carbonyl]amino}ethyl)-5chloro-1,3-thiazole-4-carboxylic acid (185) (220 mg, 0.72 mmol ), (3-chloropyridin-2-yl)methanamine dihydrochloride (154.55 mg, 0.72 mmol), DIPEA (0.26 ml, 2 mmol), HATU (0.33 g, 0.85 mmol) in admixture tetrahydrofuran (10 ml)/DMF (2 ml) gave the title compound (360 mg, quant.) as a white solid after purification by flash column chromatography (eluting with a 20-60% EtOAc/heptane gradient). The compound was used in the next step without further purification.

1H NMR (CDCl3, 500 MHz): δ [ppm] = 8.66 (s, 1H), 8.56-8.52 (m, 1H), 7.74 (dd, J=8.0 , 1.4 Hz, 1H), 7.24 (dd, J=8.0, 4.7 Hz, 1H), 4.98 (s, 1H), 4.85 (d, J=4.8 Hz , 2H), 3.61 (q, J=6.1 Hz, 2H), 3.17 (t, J=6.3 Hz, 2H), 1.46 (s, 9H).

HPLC-MS (method A): [m/z]: 431.1 [M+H]+.

tert-Butyl N-(2-{5-chloro-4-[(pyridin-2-ylmethyl)carbamoyl]-1,3-thiazol-2-yl}ethyl)carbamate (188)

Similarly, using the general procedure 6, 2-(2-{[(tert-Butoxy)carbonyl]amino}ethyl)-5chloro-1,3-thiazole-4-carboxylic acid (185) (216 mg, 0.7 mmol ), pyridin-2-ylmethanamine (0.09 ml, 0.84 mmol), DIPEA (0.26 ml, 1 mmol), HATU (0.32 g, 0.84 mmol) in tetrahydrofuran (10 ml) was stirred at room temperature. More pyridin-2-ylmethanamine (0.09 ml, 0.84 mmol), DIPEA (0.26 ml, 0 mol) and HATU (0.32 g, 0 mol) were added and the reaction continued. The title compound (227 mg, 78%) was isolated as a brown oil after purification by flash column chromatography (eluting with a 5-100% EtOAc/heptane gradient). The compound was used in the next step without further purification.

1H NMR (Methanol-d4, 250 MHz): δ [ppm] = 8.50 (d, J=4.4 Hz, 1H), 7.81 (td, J=7.8, 1.5 Hz, 1H), 7.44 (d, J=7.9 Hz, 1H), 7.36-7.27 (m, 1H), 4.73 (s, 3H), 4.68 (s, 2H ), 3.13 (t, J=6.4 Hz, 2H), 1.41 (s, 9H).

HPLC-MS (Method A): [m/z]: 397.10 [M+H] ⁺ .

tert-Butyl N-(2-{5-chloro-4-[(pyridazin-3-ylmethyl)carbamoyl]-1,3-thiazol-2-yl}ethyl)carbamate (189)

Similarly, using the general procedure 6, 2-(2-{[(tert-butoxy)carbonyl]amino}ethyl)-5chloro-1,3-thiazole-4-carboxylic acid (185) (220 mg, 0.72 mmol ), pyridazin-3-ylmethanamine (0.12 g, 1 mmol), DIPEA (0.31 ml, 2 mmol), HATU (0.41 g, 1 mmol) in DCM (20 ml) gave the title compound ( 278 mg, 91%) as a yellow residue after purification by flash column chromatography (gradient elution 50-100% EtOAc/heptane followed by 2-6% methanol/ethyl acetate). The compound was used in the next step without further purification.

HPLC-MS (method A): [m/z]: 398.05 [M+H] ⁺ .

tert-Butyl N-(2-{5-chloro-4-[(pyrimidin-2-ylmethyl)carbamoyl]-1,3-thiazol-2-yl}ethyl)carbamate (190)

Similarly, using the general procedure 6, 2-(2-{[(tert-butoxy)carbonyl]amino}ethyl)-5chloro-1,3-thiazole-4-carboxylic acid (185) (579 mg, 1.54 mmol ), 1-(pyrimidin-2-yl)methanamine (0.14 g, 1.28 mmol), DIPEA (0.67 ml, 3.85 mmol), HATU (0.585 g, 1.54 mmol) in tetrahydrofuran (10 ml)/DMF (2 ml) gave the title compound (0.483 g, 73%) as a white solid after purification by flash column chromatography (eluting with a 50-100% EtOAc/heptane gradient). The compound was used in the next step without further purification.

- 158 039916

1H NMR (DMSO-66, 500 MHz): δ [ppm] = 8.83-8.79 (m, 1H), 8.78 (d, J=4.9 Hz, 2H), 7, 41 (t, J=4.9 Hz,

1H), 7.11-7.05 (m, 1H), 4.64 (d, J=5.9 Hz, 2H), 3.09 (t, J=6.6 Hz, 2H), 1, 37 (s, 9H).

HPLC-MS (method A): [m/z]: 398 [M+H]+.

tert-Butyl N-[2-(5-chloro-4-{[(5-methylpyrimidin-2-yl)methyl]carbamoyl}-1,3-thiazol-2yl)ethyl]carbamate (191) o

Similarly, using the general procedure 6, 2-(2-{[(tert-Butoxy)carbonyl]amino}ethyl)-5chloro-1,3-thiazole-4-carboxylic acid (185) (556 mg, 1.48 mmol ), (5-methylpyrimidin-2-yl)methanamine (0.14 g, 1.14 mmol), DIPEA (0.396 ml, 2.274 mmol), HATU (0.562 g, 1.478 mmol) in tetrahydrofuran (10 ml)/DMF (2 ml) gave the title compound (577 mg, 92%) as a colorless oil after purification by flash column chromatography (eluting with a 70-100% EtOAc/heptane gradient). The compound was used in the next step without further purification.

1H NMR (DMSO-06, 500 MHz) δ 8.77 (t, J=5.8 Hz, 1H), 8.63 (s, 2H), 7.08 (s, 1H), 4.60 (d , J=5.9 Hz, 2H), 3.09 (t, J=6.6 Hz, 2H), 2.27 (s, 3H), 1.37 (s, 9H).

HPLC-MS (method A): [m/z]: 412.05 [M+H] ⁺ .

2-(2-Aminoethyl)-5-χλορ-Ν-[(3-fluoropyridin-2-yl)methyl]-1,3-thiazole-4-carboxamide (192)

Similarly, using general procedure 2, 12M HC (8 ml) and tert-butyl Ν-[2-(5-χλορ-4{[(3-fluoropyridin-2-yl)methyl]carbamoyl}-1,3-thiazole -2-yl)ethyl]carbamate (186) (1 g, 2.41 mmol) in MeOH (40 mL) at room temperature gave the title compound (703 mg, 93%) as a white solid after cartridge purification SCX-2 (gradient elution with 100% DCM followed by 100% MeOH followed by 7N NHs/MeOH).

1H NMR (DMSO-66, 500 MHz): δ [ppm] = 8.73 (t, J=5.6 Hz, 1H), 8.40 (dt, J=4.6, 1.4 Hz, 1H), 7.71 (ddd, J=10.0, 8.3, 1.2 Hz, 1H), 7.42 (dt, J=8.6, 4.4 Hz, 1H), 4 .63 (dd, J=5.7, 1.5 Hz, 2H), 3.02 (t, J=6.0 Hz, 2H), 2.93 (t, J=6.1 Hz, 2H) .

HPLC-MS (method A): [m/z]: 314.95 [M+H] ⁺ .

2-(2-Aminoethyl)-5-chloro-N-[(3-chloro-pyridin-2-yl)methyl]-1,3-thiazole-4-carboxamide (193)

Similarly, using general procedure 2, 12M HC (2 ml) and tert-butyl Ν-[2-(5-χλορ-4{[(3-chloropyridin-2-yl)methyl]carbamoyl}-1,3-thiazole -2-yl)ethyl]carbamate (187) (360 mg, 0.83 mmol) in MeOH (10 ml) at room temperature gave the title compound (320 mg, 95%) as a white solid.

1H NMR (DMSO-66, 250 MHz): δ [ppm] = 8.93 (t, J=6.0 Hz, 1H), 8.52 (dd, J=4.7, 1.3 Hz, 1H), 8.04 (s, 2H), 7.97 (dd, J=8.1, 1.4 Hz, 1H), 7.40 (dd, J=8.1, 4.7 Hz , 1H), 4.68 (d, J=5.8 Hz, 2H), 3.36-3.26 (m, 4H).

HPLC-MS (method A): [m/z]: 331.1 [M+H] ⁺ .

2-(2-Aminoethyl)-5-chloro-N-(pyridin-2-ylmethyl)-1,3-thiazole-4-carboxamide dihydrochloride (194) o

Similarly, using general procedure 2, 12M HC (2 ml) and tert-butyl Ν-(2-{5-χλορ-4[(pyridin-2-ylmethyl)carbamoyl]-1,3-thiazol-2-yl} ethyl)carbamate (188) (260 mg, 0.66 mmol) in MeOH (10 mL) at room temperature gave the title compound (198 mg, 82%) as a dark yellow gum. The resulting crude product was used without further purification.

1H NMR (Methanol-a ₄ , 250 MHz): δ [ppm] = 8.80 (d, J=5.4 Hz, 1H), 8.64 (td, J=8.0, 1, 3 Hz, 1H), 8.16 (d, J=8.1 Hz, 1H), 8.03 (t, J=6.7 Hz, 1H), 4.99 (s, 2H), 3.60 -3.39 (m, 4H).

2-(2-Aminoethyl)-5-chloro-N-(pyridazin-3-ylmethyl)-1,3-thiazole-4-carboxamide dihydrochloride (195) o ^{2 ns | N} SG ^S ^ ^NH 2

Similarly, using General Procedure 2, 12M HC (1.86 mL) and tert-butyl Ν-(2-{5-χλορ4-[(pyridazin-3-ylmethyl)carbamoyl]-1,3-thiazol-2-yl }ethyl)carbamate (189) (278 mg, 0.65 mmol) in MeOH (10 mL) at 60°C gave the crude title compound (296 mg) as a light brown residue. The compound was used in the next step without further purification.

HPLC-MS (method A): [m/z]: 298 [M+H]+.

2-(2-Aminoethyl)-5-chloro-N-(pyrimidin-2-ylmethyl)-1,3-thiazole-4-carboxamide dihydrochloride (196)

Similarly, using General Procedure 2, 12M HC (1.56 mL) and t-butyl A(2-{5-chloro4-[(pyrimidin-2-ylmethyl)carbamoyl]-1,3-thiazol-2-yl} ethyl)carbamate (190) (483 mg, 0.938 mmol) in MeOH (3 mL) at 40°C gave the title compound (563 mg, 100%) as a beige solid.

¹ H NMR (Methanol-J4, 500 MHz): δ [ppm] = 8.83 (d, J=5.0 Hz, 2H), 7.49 (t, J=5.0 Hz, 1H ), 4.82 (s, 2H), 3.49 (t, J=6.2 Hz, 2H), 3.39 (t, J=6.2 Hz, 2H).

HPLC-MS (method A): [m/z]: 297.9 [M+H] ⁺ .

2-(2-Aminoethyl)-5-chloro-N-[(5-methylpyrimidin-2-yl)methyl]-1,3-thiazole-4-carboxamide dihydrochloride (197)

Similar to general procedure 2, 12M HC (1.74 ml) and tert-butyl Ν-[2-(5-χλορ-4-{[(5methylpyrimidin-2-yl)methyl]carbamoyl}-1,3-thiazol-2 -yl)ethyl]carbamate (191) (580 mg, 1.04 mmol) in MeOH (6 ml) at room temperature gave the title compound (447 mg, 100%) as a white solid.

¹ H NMR (Methanol-J4, 500 MHz): δ [ppm] = 8.74 (s, 2H), 4.80 (s, 2H), 3.49 (t, J=6.2 Hz , 2H), 3.38 (t, J=6.2 Hz, 2H), 2.39 (s, 3H).

HPLC-MS (Method A): [m/z]: 311.95 [M+H] ⁺ .

2-{2-[(1H-1,3-Benzodiazol-2-ylmethyl)amino]ethyl}-5-chloro-N-[(3-fluoropyridin-2-yl)methyl]-1,3thiazol-4-carboxamide (illustrative example #151)

Similar to general procedure 3, 2-(2-Aminoethyl)-5-chloro-N-[(3-fluoropyridin-2-yl)methyl]-1,3-thiazole4-carboxamide dihydrochloride (192) (143 mg, 0.37 mmol), 1H-1,3-benzodiazol-2-carbaldehyde (70.1 mg, 0.48 mmol), MgSO ₄ (200 mg) in MeOH (10 ml) at room temperature for 3 days, with the addition of NaBH ₄ (28 mg, 0.74 mmol) gave the title compound (94 mg, 56%) as a white solid after purification by preparative HPLC.

1H NMR (DMSO-a ₆ , 500 MHz): δ [ppm] = 12.19 (s, 1H), 8.71 (t, J=5.6 Hz, 1H), 8.37 (dt , J=4.6, 1.3 Hz, 1H), 7.70 (ddd, J=10.0, 8.4, 1.1 Hz, 1H), 7.54 (d, J=7.2 Hz, 1H), 7.45 (d, J=7.0 Hz, 1H), 7.40 (dt, J=8.6, 4.4 Hz, 1H), 7.14 (t, J=7 .5 Hz, 2H), 4.66-4.58 (m, 2H), 3.97 (s, 2H), 3.12 (t, J=6.3 Hz, 2H), 2.94 (t , J=6.3 Hz, 2H).

HPLC-MS (method A): [m/z]: 445.1 [M+H] ⁺ .

2-{2-[(1H-1,3-Benzodiazol-2-ylmethyl)amino]ethyl}-5-chloro-N-[(3-chloropyridin-2-yl)methyl]-1,3thiazol-4-carboxamide (illustrative example #159)

Similar to general procedure 3, 2-(2-Aminoethyl)-5-chloro-N-[(3-chloropyridin-2-yl)methyl]-1,3-thiazole4-carboxamide dihydrochloride (193) (320 mg, 0.79 mmol), 1H-1,3-benzodiazol-2-carbaldehyde (127.3 mg, 0.87 mmol), DIPEA (0.32 ml, 2.0 mmol) and MgSO ₄ (300 mg) in MeOH (20 ml ) at room temperature for 16 h followed by addition of NaBH ₄ (60 mg, 1.59 mmol) gave the title compound (191 mg, 52%) as a white solid after purification by preparative HPLC.

¹ H NMR (DMSO-J6, 500 MHz): δ [ppm] = 12.18 (s, 1H), 8.72 (t, J=5.6 Hz, 1H), 8.49 (dd , J=4.7, 1.3 Hz, 1H), 7.95 (dd, J=8.1, 1.3 Hz, 1H), 7.54 (s, 1H), 7.46 (s, 1H), 7.38 (dd, J=8.1, 4.7 Hz, 1H), 7.17-7.09 (m, 2H), 4.65 (d, J=5.5 Hz, 2H ), 3.98 (d, J=4.1 Hz, 2H), 3.13 (t, J=6.3 Hz, 2H), 2.95 (q, J=6.1, 5.3 Hz , 2H), 2.74 (s, 1H).

HPLC-MS (method C): [m/z]: 461 [M+H] ⁺ .

2-{2-[(1H-1,3-Benzodiazol-2-ylmethyl)amino]ethyl}-5-chloro-N-(pyridin-2-ylmethyl)-1,3-thiazole-4carboxamide (Illustrative Example No. 160 )

- 160 039916

Similar to general procedure 3, 2-(2-aminoethyl)-5-chloro-N-(pyridin-2-ylmethyl)-1,3-thiazole-4-carboxamide dihydrochloride (194) (190 mg, 0.51 mmol), 1H- 1,3-benzodiazol-2-carbaldehyde (113 mg, 0.77 mmol), DIPEA (0.3 ml, 2.0 mmol) and MgSO ₄ (400 mg) in DCM (10 ml) and MeOH (10 ml) at room temperature for 16 hours, followed by addition of NaBH ₄ (39 mg, 1.03 mmol) gave the title compound (60 mg, 27%) as a pale yellow solid after purification by preparative HPLC.

1H NMR (DMSO-J6, 500 MHz): δ [ppm] = 12.19 (s, 1H), 8.90 (t, J=6.0 Hz, 1H), 8.50 (d, J=5.5Hz, 1H), 7.75(td, J=7.7, 1.8Hz, 1H), 7.58-7.42(m, 2H), 7.31(d, J = 7.8 Hz, 1H), 7.27 (dd, J=7.0, 5.3 Hz, 1H), 7.17-7.10 (m, 2H), 4.54 (d, J= 6.0 Hz, 2H), 3.98 (s, 2H), 3.12 (t, J=6.3 Hz, 2H), 2.95 (t, J=6.3 Hz, 2H), 2 .76 (s, 1H).

HPLC-MS (method C): [m/z]: 427.1 [M+H] ⁺ .

5-Chloro-N-(pyridin-2-ylmethyl)-2-(2-{1,8,12-triazatricyclo[7.4,O.0 ² .7]trideca-2(7),3.5, 8-tetraen12-yl}ethyl)-1,3-thiazole-4-carboxamide (illustrative example No. 189)

Similar to general procedure 8, 2-(2-aminoethyl)-5-chloro-N-(pyridin-2-ylmethyl)-1,3-thiazole-4-carboxamide dihydrochloride (194) (361.5 mg, 0.98 mmol), N-(2-nitrophenyl)prop-2-enamide (D) (187.9 mg, 0.98 mmol), DBU (219 µl, 1.47 mmol) in MeCN (5 ml) at room temperature for 3 h gave a mixture of mono- and bis-alkylated intermediates, which were separated by flash column chromatography (gradient elution 0-10% MeOH/EtOAc, then 0-10% MeOH/DCM). Monoalkylated intermediate (5-chloro-2-[2-({2-[(2-nitrophenyl)carbamoyl]ethyl}amino)ethyl]-1h[-(pyridin-2ylmethyl)-1,3-thiazole-4-carboxamide) (66.0 mg, 0.135 mmol) was further reacted with iron powder (15.1 mg, 0.270 mmol) in AcOH (1 mL) at 80°C for 1 hour to give the title compound (61 mg, 25%) as a brown oil after purification by flash column chromatography (eluting with a 0-10% MeOH/DCM gradient).

¹ H NMR (Acetone-J6, 500 MHz): δ [ppm] = 8.53 (d, J=4.2 Hz, 2H), 7.76 (td, J=7.7, 1, 8 Hz, 1H), 7.61-7.53 (m, 1H), 7.45-7.37 (m, 2H), 7.26 (dd, J=7.0, 4.9 Hz, 1H ), 7.22-7.14 (m, 2H), 5.14 (s, 2H), 4.66 (d, J=5.7 Hz, 2H), 3.39 (t, J=6, 2 Hz, 2H), 3.36-3.28 (m, 2H), 3.20 (t, J=6.3 Hz, 2H), 3.14 (t, J=6.2 Hz, 2H) .

HPLC-MS (method B): [m/z]: 453.1 [M+H]+.

2-{2-[(1H-1,3-Benzodiazol-2-ylmethyl)amino]ethyl}-5-chloro-N-(pyridazin-3-ylmethyl)-1,3-thiazole-4carboxamide (Illustrative Example No. 161 )

Similar to general procedure 3, 2-(2-aminoethyl)-5-chloro-N-(pyridazin-3-ylmethyl)-1,3-thiazol-4-carboxamide dihydrochloride (195) (296 mg, 0.8 mmol), 1H- 1,3-benzodiazol-2-carbaldehyde (151.7 mg, 1.04 mmol), DIPEA (0.42 ml, 2.0 mmol) and MgSO ₄ (400 mg) in DCM (10 ml) and MeOH (10 ml) at room temperature for 3 days, adding NaBH ₄ (60 mg, 1.59 mmol) gave the title compound (96 mg, 28%) as a white solid after purification by preparative HPLC.

1H NMR (DMSO-J6, 500 MHz): δ [ppm] = 12.18 (s, 1H), 9.14 (dd, J=4.8, 1.5 Hz, 1H), 9, 07 (t, J=6.1Hz, 1H), 7.66 (dd, J=8.5, 4.8Hz, 1H), 7.60 (dd, J=8.5, 1.6Hz , 1H), 7.54 (d, J=7.4 Hz, W), 7.45 (d, J=7.7 Hz, 1H), 7.13 (n, J=7.0, 6, 6 Hz, 2H), 4.72 (d, J=6.1 Hz, 2H), 3.97 (s, 2H), 3.12 (t, J=6.3 Hz, 2H), 2.95 (t, J=6.2 Hz, 2H), 2.75 (s, 1H).

HPLC-MS (method C): [m/z]: 428 [M+H] ⁺ .

2-{2-[(1H-1,3-Benzodiazol-2-ylmethyl)amino]ethyl}-5-chloro-N-(pyrimidin-2-ylmethyl)-1,3-thiazole-4carboxamide (Illustrative Example No. 175 )

Similar to general procedure 3, 2-(2-aminoethyl)-5-chloro-N-(pyrimidin-2-ylmethyl)-1,3-thiazol-4- 161 039916 carboxamide dihydrochloride (196) (563 mg, 0.94 mmol ), 1H-benzimidazole-2-carbaldehyde (178 mg, 1.22 mmol), DIPEA (0.654 ml, 0.74 mmol) in MeOH (15 ml) at room temperature for 24 h, followed by addition of NaBH4 (53 mg , 1.41 mmol) gave the title compound (224 mg,

54%) as a white solid after purification by basic prep-HPLC.

¹ H NMR (DMSO-d6, 500 MHz): δ [ppm] = 12.20 (br s, 1H), 8.80-8.74 (m, 3H), 7.50 (br. s, 2H), 7.40 (t, J=4.9 Hz, 1H), 7.21-7.07 (m, 2H), 4.64 (d, J=5.9 Hz, 2H), 3.98 (s, 2H), 3.13 (t, J=6.3 Hz, 2H), 2.96 (t, J=6.3 Hz, 2H), 2.76 (br s, 1H ).

HPLC-MS (method d): [m/z]: 428.2 [M+H]+.

2-{2-[(1H-1,3-Benzodiazol-2-ylmethyl)amino]ethyl}-5-chloro-N-[(5-methylpyrimidin-2-yl)methyl]-1,3thiazol-4-carboxamide (illustrative example #178)

Similar to general procedure 3, 2-(2-Aminoethyl)-5-chloro-N-[(5-methylpyrimidin-2-yl)methyl]-1,3thiazole-4-carboxamide dihydrochloride (197) (223 mg, 0.52 mmol), 1H-benzimidazole-2-carbaldehyde (99 mg, 0.68 mmol), DIPEA (0.363 ml, 2.09 mmol) and MgSO ₄ (400 mg) in MeOH (6 ml) at room temperature for 24 h , followed by addition of NaBH ₄ (30 mg, 0.78 mmol) gave the title compound (120 mg, 52%) as a white solid after purification by basic pre-HPLC.

1H NMR (DMSO-d6, 500 MHz): δ [ppm] = 12.21 (s, 1H), 8.72 (t, J=5.8 Hz, 1H), 8.63-8, 57 (m, 2H), 7.50 (s, 1H), 7.14 (dd, J=6.0, 2.7 Hz, 2H), 4.61-4.56 (m, 2H), 3 .98 (s, 2H), 3.13 (t, J=6.3 Hz, 2H), 2.95 (t, J=6.3 Hz, 2H), 2.75 (s, 1H), 2 .26 (s, 3H).

HPLC-MS (method C): [m/z]: 442 [M+H] ⁺ .

5-Chloro-N-[(3-fluoropyridin-2-yl)methyl]-2-[2-({2-[(2-nitrophenyl)carbamoyl]ethyl}amino)ethyl]-1,3thiazole-4-carboxamide (198)

Similar to general procedure 8, 2-(2-aminoethyl)-5-chloro-N-[(3-fluoropyridin-2-yl)methyl]-1,3-thiazole4-carboxamide (192) (700 mg, 2.22 mmol ) and N-(2-nitrophenyl)prop-2-enamide (D) (470 mg, 2.45 mmol) were combined in MeCN (40 ml) and DBU (0.37 ml, 2.0 mmol) was added. The mixture was stirred at room temperature for 3 days. The reaction mixture was applied directly to the silica gel. Purification by flash column chromatography (eluting with a 0-10% MeOH/DCM gradient) gave the title compound (390 mg, 25%) as a yellow solid.

HPLC-MS (method A): [m/z]: 507 [M+H] ⁺ .

2-(2-{[2-(1H-1,3-Benzodiazol-2-yl)ethyl]amino}ethyl)-5-chloro-N-[(3-fluoropyridin-2-yl)methyl]-1, 3thiazole-4-carboxamide trihydrochloride (illustrative example No. 154)

Similar to the general procedure 8, 5-Chloro-N-[(3-fluoropyridin-2-yl)methyl]-2-[2-({2-[(2nitrophenyl)carbamoyl]ethyl}amino)ethyl]-1,3- thiazole-4-carboxamide (198) (390 mg, 0.77 mmol) and iron powder (172 mg, 3 mmol) in AcOH (5 ml) at 90° C. for 30 min gave the crude product. The resulting residue was flash chromatographed through a pad of silica gel eluting with a gradient of 0.5-40% MeOH/DCM followed by 20% 7M NH3 in MeOH/DCM followed by basic prep-HPLC purification to give the title product as the free base. The resulting residue was redissolved in MeOH (10 ml) and 12M HC (1 ml) was added for 1 hour. The solvent was evaporated in vacuo to give the title compound as a pale yellow solid.

1H NMR (D ₂ O, 500 MHz): δ [ppm] = 8.33-8.30 (m, 1H), 7.87 (ddd, J=9.6, 8.6, 1, 2 Hz, 1H), 7.71 (dt, J=6.8, 3.4 Hz, 2H), 7.59 (dt, J=7.5, 4.2 Hz, 1H), 7.55 ( dt, J=6.3, 3.4 Hz, 2H), 3.72-3.62 (m, 6H), 3.44 (t, J=6.6 Hz, 2H). The benzyl CH2 signal overlaps with the D2O signals.

HPLC-MS (method A): [m/z]: 459.1 [M+H] ⁺ .

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2-(2-{[2-(1H-1,3-Benzodiazol-2-yl)ethyl]amino}ethyl)-5-χΛορ-Ν-[(5-methylpyrimidin-2-yl)methyl] 1.3 -thiazole-4-carboxamide (illustrative example No. 182)

Similarly, using the general procedure 8, 2-(2-Aminoethyl)-5-χlor-N-[(5methylpyrimidin-2-yl)methyl]-1,3-thiazole-4-carboxamide dihydrochloride (197) (0.223 g, 0.522 mmol), N(2-nitrophenyl)prop-2-enamide (D) (0.090 g, 0.47 mmol) and DBU (0.171 ml, 1.148 mmol) in MeCN (10 ml) gave the intermediate (355 mg, 0. 46 mmol) which was reacted with iron powder (104 mg, 1.86 mmol) in AcOH (5 ml) at 80°C for 2.5 h to give the title compound (6 mg, 3%) as no off-white solid after purification by basic prep-HPLC, followed by flash column chromatography (eluting with a 0-5% MeOH/DCM gradient).

1H NMR (DMSO-a ₄ , 500 MHz): δ [ppm] = 8.54 (s, 2H), 7.47-7.41 (m, 2H), 7.20-7.14 ( m, 2H), 4.62 (s, 2H), 3.23-3.16 (m, 4H), 3.15-3.10 (m, 4H), 2.28 (s, 3H).

HPLC-MS (Method D): [m/z]: 456.2 [M+H]+.

2-(2-{[2-(1H-1,3-Benzodiazol-2-yl)ethyl]amino}ethyl)-5-χlor-N-(pyridazin-3-ylmethyl)-1,3-thiazole-4carboxamide (illustrative example #183)

Similarly, using the general procedure 8, 2-(2-Aminoethyl)-5-χlor-N-(pyridazin-3ylmethyl)-1,3-thiazole-4-carboxamide dihydrochloride (195) (255 mg, 0.689 mmol), N -(2-nitrophenyl)prop2-enamide (D) (132 mg, 0.689) and DBU (0.154 ml, 1.03 mmol) in MeCN (5 ml) gave an intermediate (112 mg, 0.16 mmol) which reacted with iron powder (18 mg, 0.33 mmol) in AcOH (1 mL) at 80°C for 2.5 h to give the title compound (6.4 mg, 8.5%) as a yellow oil after purification flash column chromatography (0-20% MeOH/DCM gradient elution), followed by basic prep-HPLC.

1H NMR (Methanol-d ₄ , 500 MHz): δ [ppm] = 9.08 (dd, J=4.3, 2.3 Hz, 1H), 7.72-7.63 (m, 2H), 7.47 (dd, J=5.9, 3.2 Hz, 2H), 7.23-7.15 (m, 2H), 4.80 (s, 2H), 3.16-3 .02 (m, 8H).

HPLC-MS (method B): [m/z]: 440.4 [M-H] ^- .

2-(2-{[2-(1H-1,3-Benzodiazol-2-yl)ethyl]amino}ethyl)-5-χlor-N-(pyridin-2-ylmethyl)-1,3-thiazole-4carboxamide (illustrative example #184)

Similarly, using the general procedure 8, 2-(2-Aminoethyl)-5-χlor-N-(pyridine-2ylmethyl)-1,3-thiazole-4-carboxamide dihydrochloride (194) (361.6 mg, 0.978 mmol) , N-(2-nitrophenyl)prop2-enamide (D) (187.9 mg, 0.978 mmol), DBU (0.219 ml, 1.467 mmol) in MeCN (5 ml) gave the intermediate (180 mg, 0.26 mmol), which was reacted with iron powder (29 mg, 0.54 mmol) in AcOH (1 ml) at 80°C for 2 h to give the title compound (8.8 mg, 7.5%) as a yellow oil after purification basic prep-HPLC method.

1H NMR (Methanol-d ₄ , 500 MHz): δ [ppm] = 8.47 (ddd, J=5.0, 1.6, 0.9 Hz, 1H), 7.77 (td, J=7.7, 1.8 Hz, 1H), 7.47 (dd, J=5.9, 3.2 Hz, 2H), 7.37 (d, J=7.9 Hz, 1H), 7.33-7.25(m, 1H), 7.20-7.16(m, 2H), 4.61(s, 2H), 3.16-3.01(m, 8H).

HPLC-MS (method B): [m/z]: 439.3 [M-H] ^- .

Ethyl 5-bromo-2-(2-{[(tert-butoxy)carbonyl]amino}ethyl)-1,3-thiazole-4-carboxylate (199)

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Ethyl 2-(2-{[(tert-butoxy)carbonyl]amino}ethyl)-1,3-thiazole-4-carboxylate (1) (1.56 g, 5.19 mmol) and 1,2-dibromo- 1,1,2,2-tetrachloroethane (1.86 g, 10.0 mmol) was dissolved in THF (60 ml) and cooled to -78°C. 2M NaHMDS (5.45 ml) was added dropwise and the resulting mixture was stirred at -78°C for 2 h. The reaction was quenched with saturated aqueous NH ₄ Cl (30 ml) and left to warm to room temperature. The resulting mixture was extracted with dichloromethane (3x100 ml) and the combined organic extracts were dried (Na ₂ SO ₄ ), filtered and evaporated in vacuo. Purification by flash column chromatography eluting with a 10-50% EtOAc/heptane gradient gave the title compound (1.62 g, 80%) as a colorless oil.

¹ H NMR (CDCl ₃ , 250 MHz): δ [ppm] = 4.91 (s, 1H), 4.46 (q, J=7.1 Hz, 2H), 3.56 (q, J=6.3 Hz, 2H), 3.21 (t, J=6.3 Hz, 2H), 1.50-1.42 (m, 12H).

HPLC-MS (method A): [m/z]: 400.9/402.85 [M+H] ⁺ .

2-(2-{[(tert-Butoxy)carbonyl]amino}ethyl)-5-(prop-1-en-2-yl)-1,3-thiazole-4-carboxylate (200)

Ethyl 5-bromo-2-(2-{[(tert-butoxy)carbonyl]amino}ethyl)-1,3-thiazole-4-carboxylate (199) (626 mg, 1.65 mmol), PdCl ₂ (dppf (120 mg, 0.17 mmol) and K ₂ CO ₃ (460 mg, 3.0 mmol) were dissolved in MeCN (18 ml) and water (18 ml) under nitrogen atmosphere and 4,4,5,5-tetramethyl was added -2-(prop-1-en-2-yl)-1,3,2-dioxaborolane (0.38 ml, 1.98 mmol) The resulting mixture was heated at 80°C for 30 min, cooled to room temperature and diluted with water The resulting mixture was extracted with dichloromethane (3x80 ml) and the combined organic extracts were dried (Na ₂ SO ₄ ), filtered and evaporated in vacuo Purification by flash column chromatography eluting with a gradient of 10-60% EtOAc/heptane gave the indicated the title compound (558 mg, 99%) as a yellow oil.

Ή NMR (CDCl ₃ , 250 MHz): δ [ppm] = 5.23 (s, 1H), 5.12 (s, 1H), 4.94 (s, 1H), 4.32 (q , J=7.2Hz, 2H), 3.47(q, J=6.5Hz, 2H), 3.09(t, J=6.5Hz, 2H), 2.07(s, 3H ), 1.37 (s, 9H), 1.31 (t, J=7.2 Hz, 3H).

HPLC-MS (Method A): [m/z]: 341.05 [M+H]+.

Ethyl 2-(2-{[(tert-butoxy)carbonyl]amino}ethyl)-5-(trifluoromethyl)-1,3-thiazole-4-carboxylate (201)

Ethyl 5-bromo-2-(2-{[(tert-butoxy)carbonyl]amino}ethyl)-1,3-thiazole-4-carboxylate (199) (0.54 g, 1.43 mmol) was dissolved in DMF (30 ml) under nitrogen atmosphere and copper (I) iodide (1.36 g, 7.0 mmol), triphenylarsan (0.14 ml, 0.57 mmol), Pd ₂ cdba ₃ (0.065 g, 0.07 mmol) and methyl difluoro(fluorosulfonyl)acetate (0.23 ml, 1.85 mmol). The resulting mixture was heated at 100°C for 3 h, then cooled to room temperature and evaporated in vacuo. The resulting residue was partitioned between EtOAc and saturated aqueous NaHCO ₃ . The phases were separated, the aqueous phase was extracted with ethyl acetate (2x80 ml), and the combined organic extracts were washed with saturated sodium chloride solution (2x50 ml), dried (Na ₂ SO ₄ ), filtered and evaporated in vacuum. Purification by flash column chromatography (eluting with a 10-60% EtOAc/heptane gradient) gave the title compound (0.447 g, 84%) as a yellow oil.

1H NMR (DMSO-d ₆ , 500 MHz): δ [ppm] = 4.92 (s, 1H), 4.48 (q, J=7.1 Hz, 2H), 3.60 (q , J=6.2 Hz, 2H), 3.27 (t, J=6.2 Hz, 2H), 1.46 (s, 9H), 1.44 (t, J=7.1 Hz, 3H ).

HPLC-MS (method A): [m/z]: 391 [M+Na]+.

2-(2-{[(tert-Butoxy)carbonyl]amino}ethyl)-5-(prop-1-en-2-yl)-1,3-thiazole-4-carboxylic acid (202)

Similar to general procedure 5, ethyl 2-(2-{[(tert-butoxy)carbonyl]amino}ethyl)-5-(prop-1-en-2yl)-1,3-thiazole-4-carboxylate (200) ( 550 mg, 1.62 mmol) and LiOH (390 mg, 16.0 mmol) in THF (20 mL) and water (20 mL) gave the crude title compound (569 mg, quant) as a yellow oil.

¹ H NMR (CDCl ₃ , 500 MHz): δ [ppm] = 5.41 (s, 1H), 5.38 (s, 1H), 4.84 (s, 1H), 3.59 ( m, 2H), 3.16 (t, J=6.4 Hz, 2H), 2.23 (s, 3H), 1.47 (s, 9H).

HPLC-MS (method A): [m/z]: 312.9 [M+H] ⁺ .

2-(2-{[(tert-Butoxy)carbonyl]amino}ethyl)-5-(trifluoromethyl)-1,3-thiazole-4-carboxylic acid (203)

- 164 039916

Similar to general procedure 5, ethyl 2-(2-{[(tert-butoxy)carbonyl]amino}ethyl)-5-(trifluoromethyl)1,3-thiazole-4-carboxylate (201) (447 mg, 1.21 mmol ) and LiOH (320 mg, 13.0 mmol) in THF (15 ml) and water (15 ml) gave the title compound (365 mg, 88%) as a pale yellow solid.

¹ H NMR (CDCl ₃ , 500 MHz): δ [ppm] = 4.81 (s, 1H), 3.53 (q, J=6.0 Hz, 2H), 3.18 (br. s, 2H), 1.37 (s, 9H).

HPLC-MS (method A): [m/z]: 362.95 [M+Na]+.

tert-Butyl

N-[2-(4-{[(3-Fluoropyridin-2-yl)methyl]carbamoyl}-5-(prop-1-en-2-yl)-1,3-thiazol-2yl)ethyl]carbamate ( 204)

Similar to general procedure 6, 2-(2-{[(tert-butoxy)carbonyl]amino}ethyl)-5-(prop-1-en-2-yl)-1,3thiazole-4-carboxylic acid (202) ( 569 mg, 1.82 mmol), (3-fluoropyridin-2-yl)methanamine dihydrochloride (A2) (435 mg, 2.19 mmol), DIPEA (1.11 ml, 6.0 mmol) and HATU (830 mg , 2.0 mmol) in DCM (20 mL) gave the title compound (544 mg, 71%) as a colorless oil after purification by flash column chromatography (eluting with a 0-60% EtOAc/heptane gradient).

1H NMR (CDCl ₃ , 500 MHz): δ [ppm] = 8.51-8.43 (m, 2H), 7.45-7.38 (m, 1H), 7.30-7, 24 (m, 1H), 5.34 (m, 1H), 5.27 (s, 1H), 4.83 (dd, J=5.1, 1.6 Hz, 2H), 3.61 (m , 2H), 3.15 (m, 2H), 2.25 (s, 3H), 1.46 (s, 9H).

HPLC-MS (method A): [m/z]: 421.05 [M+H]+.

tert-Butyl N-[2-(4-{[(3-fluoropyridin-2-yl)methyl]carbamoyl}-5-(trifluoromethyl)-1,3-thiazol-2yl)ethyl]carbamate (205)

Similar to general procedure 6, 2-(2-{[(tert-butoxy)carbonyl]amino}ethyl)-5-(trifluoromethyl)-1,3thiazole-4-carboxylic acid (203) (365 mg, 1.07 mmol) , (3-fluoropyridin-2-yl)methanamine dihydrochloride (A2) (256.2 mg, 1.29 mmol), DIPEA (0.65 ml, 4 mmol) and HATU (490 mg, 1.3 mmol) in DCM (20 ml) gave the title compound (344 mg, 72%) after purification by flash column chromatography (eluting with a 20-30% EtOAc/heptane gradient).

1 H NMR (CDCl ₃ , 250 MHz): δ [ppm] = 8.49 (s, 1H), 8.35 (dt, J=4.6, 1.3 Hz, 1H), 7.35 (ddd, J=9.4, 8.4, 1.3 Hz, 1H), 7.25-7.15 (m, 1H), 4.85 (s, 1H), 4.77 (dd, J =5.0, 1.6 Hz, 2H), 3.54 (kv, J=6.3 Hz, 2H), 3.17 (t, J=6.3 Hz, 2H), 1.37 (s , 9H).

HPLC-MS (method A): [m/z]: 449.05 [M+H] ⁺ .

2-(2-Aminoethyl)-N-[(3-fluoropyridin-2-yl)methyl]-5-(prop-1-en-2-yl)-1,3-thiazol-4-carboxamide dihydrochloride (206)

Similar to general procedure 2, tert-butyl N-[2-(4-{[(3-fluoropyridin-2-yl)methyl]carbαmoyl}-5(prop-1-en-2-yl)-1,3-thiazole -2-yl)ethyl]carbamate (204) (544 mg, 1.29 mmol) and 12M HC (2 ml) in MeOH (10 ml) gave the title compound (486 mg, 94%) as a white solid .

HPLC-MS (method A): [m/z]: 321 [M+H]+.

2-(2-Aminoethyl)-N-[(3-fluoropyridin-2-yl)methyl]-5-(trifluoromethyl)-1,3-thiazole-4-carboxamide dihydrochloride (207)

Similar to general procedure 2, tert-butyl N-[2-(4-{[(3-fluoropyridin-2-yl)methyl]carbαmoyl}-5(trifluoromethyl)-1,3-thiazol-2-yl)ethyl]carbamate (205) (347 mg, 0.77 mmol) and 12M HC (2 ml) in MeOH (10 ml) gave the title compound (330 mg, quant) as a white solid.

1H NMR (DMSO-d6, 250 MHz): δ [ppm] = 9.22 (t, J=5.6 Hz, 1H), 8.40 (dt, J=4.6, 1.4 Hz, 1H), 8.16 (s, 3H), 7.74 (ddd, J=10.1, 8.4, 1.4 Hz, 1H), 7.43 (dt, J=8.4, 4.4 Hz, 1H), 4.68 (dd, J=5.8, 1.5 Hz, 2H), 3.42-3.22 (m, 4H).

- 165 039916

HPLC-MS (method A): [m/z]: 349 [M+H]+.

2-{2-[(1H-1,3-Benzodiazol-2-ylmethyl)amino]ethyl}-N-[(3-fluoropyridin-2-yl)methyl]-5-(prop-1-en-2yl) -1,3-thiazole-4-carboxamide (illustrative example No. 177)

Similarly, using the general procedure 3, 2-(2-aminoethyl)-N-[(3-fluoropyridin-2yl)methyl]-5-(prop-1-en-2-yl)-1,3-thiazol-4 -carboxamide dihydrochloride (206) (260 mg, 0.66 mmol), 1Hbenzimidazole-2-carbaldehyde (96.6 mg, 0.66 mmol), DIPEA (0.36 ml, 2.0 mmol) and MgSO ₄ (300 mg) in MeOH (20 mL) at room temperature for 18 h followed by addition of NaBH ₄ (500 mg, 1.3 mmol) gave the title compound (148 mg, 50%) as a light yellow solid after purification by basic prep-HPLC.

¹ H NMR (DMSO-d6, 500 MHz): δ [ppm] = 12.19 (s, 1H), 8.67 (t, J=5.5 Hz, 1H), 8.36 (d , J=4.6 Hz, 1H), 7.69 (t, J=8.8 Hz, 1H), 7.58-7.43 (m, 2H), 7.40 (m, 1H), 7 .16-7.09 (m, 2H), 5.21 (s, 1H), 5.15 (s, 1H), 4.61 (d, J=5.5 Hz, 2H), 3.97 ( s, 2H), 3.11 (t, J=6.7 Hz, 2H), 2.96 (t, J=6.7 Hz, 2H), 2.08 (s, 3H).

HPLC-MS (method C): [m/z]: 451.1 [M+H] ⁺ .

2-{2-[(1H-1,3-Benzodiazol-2-ylmethyl)amino]ethyl}-N-[(3-fluoropyridin-2-yl)methyl]-5(trifluoromethyl)-1,3-thiazol- 4-carboxamide (illustrative example No. 176)

Similarly, using the general procedure 3, 2-(2-aminoethyl)-N-[(3-fluoropyridin-2yl)methyl]-5-(trifluoromethyl)-1,3-thiazole-4-carboxamide dihydrochloride (207) (130 mg, 0.31 mmol), 1Hbenzimidazole-2-carbaldehyde (67.6 mg, 0.46 mmol), DIPEA (0.16 ml, 0.93 mmol) and MgSO ₄ (300 mg) in MeOH (10 ml) at room temperature for 16 h followed by addition of NaBH ₄ (23 mg, 0.62 mmol) gave the title compound (81 mg, 55%) as a light yellow solid after purification by basic prep-HPLC.

1H NMR (DMSO-d ₆ , 500 MHz): δ [ppm] = 12.21 (s, 1H), 8.96 (t, J=5.7 Hz, 1H), 8.37 (dt , J=4.6, 1.3 Hz, 1H), 7.71 (ddd, J=10.0, 8.3, 1.3 Hz, 1H), 7.53 (s, 1H), 7, 45 (s, 1H), 7.41 (dt, J=8.3, 4.4 Hz, 1H), 7.14 (s, 2H), 4.65 (dd, J=5.7, 1, 4 Hz, 2H), 3.99 (s, 2H), 3.22 (t, J=6.3 Hz, 2H), 2.99 (t, J=6.3 Hz, 2H), 2.80 (s, 1H).

HPLC-MS (Method C): [m/z]: 479.1 [M+H] ⁺ .

2-{2-[(1H-1,3-Benzodiazol-2-ylmethyl)amino]ethyl}-N-[(3-fluoropyridin-2-yl)methyl]-5-(propan-2yl)-1,3 -thiazole-4-carboxamide (illustrative example No. 180)

2-{2-[(1H-1,3-benzodiazol-2-ylmethyl)amino]ethyl}-N-[(3-fluoropyridin-2-yl)methyl]-5-(prop-1-en-2yl) -1,3-thiazole-4-carboxamide (illustrative example No. 177) (120 mg, 0.27 mmol) and palladium on carbon (10%, 28 mg, 0.027 mmol) were combined in EtOH (10 ml) and the resulting mixture stirred under hydrogen atmosphere for 1.5 hours. The reaction mixture was filtered through a pad of celite and the resulting residue was washed with MeOH. The combined filtrates were evaporated in vacuo and purified by basic prep-HPLC to give the title compound (68 mg, 56%) as a white solid.

1H NMR (DMSO-66, 500 MHz): δ [ppm] = 12.20 (s, 1H), 8.60 (t, J=5.6 Hz, 1H), 8.38-8, 34 (m, 1H), 7.70 (m, 1H), 7.58-7.43 (m, 2H), 7.40 (dt, J=8.6, 4.4 Hz, 1H), 7 .16-7.09 (m, 2H), 4.63 (dd, J=5.6, 1.4 Hz, 2H), 4.24 (hept, J=6.8 Hz, 1H), 3, 96 (s, 2H), 3.11 (t, J=6.8 Hz, 2H), 2.95 (t, J=6.8 Hz, 2H), 1.21 (d, J=6.8 Hz, 6H).

HPLC-MS (Method C): [m/z]: 453.2 [M+H] ⁺ .

2-(2-{[2-(1H-1,3-Benzodiazol-2-yl)ethyl]amino}ethyl)-X-[(3-fluoropyridin-2-yl)methyl]-5(trifluoromethyl)-1 ,3-thiazole-4-carboxamide (illustrative example No. 186)

- 166 039916

Similarly, using the general procedure 8, 2-(2-aminoethyl)-N-[(3-fluoropyridine-2yl)methyl]-5-(trifluoromethyl)-1,3-thiazole-4-carboxamide dihydrochloride (207) (242 mg, 0.57 mmol), N-(2nitrophenyl)prop-2-enamide (110.4 mg, 0.57 mmol) and DBU (0.301 ml, 2 mmol) in MeCN (10 ml) gave a crude intermediate which reacted with iron powder (56 mg, 1 mmol) in AcOH (3 mL) to give the title compound (4 mg, 2%) after two purifications by basic prep-HPLC and a final purification by flash column chromatography (gradient elution 0-20% MeOH/DCM).

1H NMR (DMSO-J6, 500 MHz): δ [ppm] = 12.14 (s, 1H), 8.94 (t, J=5.5 Hz, 1H), 8.39 (dt, J=4.5, 1.4 Hz, 1H), 7.71 (ddd, J=9.9, 8.3, 1.4 Hz, 1H), 7.41 (m, 2H), 7.10 (m, 2H), 4.64 (dd, J=5.7, 1.4 Hz, 2H), 3.18 (t, J=6.1 Hz, 2H), 3.06 (t, J= 6.1 Hz, 2H), 3.02-2.92 (m, 4H).

HPLC-MS (Method C): [m/z]: 493.1 [M+H] ⁺ .

2-(Ethoxycarbonyl)-1,3-thiazole-4-carboxylic acid (216)

3-Bromo-2-oxopropanoic acid and ethyl amino(thioxo)acetate in THF (100 ml) were stirred at 60° C. for 16 hours. The reaction mixture was evaporated in vacuo to give an orange solid. The resulting product was triturated in Et ₂ O, filtered and dried in vacuo to give the title compound (4.54 g, 62.8%) as a colorless solid.

1H NMR (DMSO-d6, 500 MHz): δ [ppm] = 13.40 (s, 1H), 8.77 (s, 1H), 4.40 (q, J=7.1 Hz, 2H), 1.35 (t, J=7.1 Hz, 3H).

HPLC-MS (ESI+): [m/z]: 201.90 [M+H]+.

Ethyl 4-(hydroxymethyl)-1,3-thiazole-2-carboxylate (217)

Isobutyl chloroformate (1.16 ml, 8.95 mmol) was added to an ice-cold (0°C) suspension of 2(ethoxycarbonyl)-1,3-thiazole-4-carboxylic acid (216) (1.5 g, 7.46 mmol ) and TEA (1.25 ml, 8.95 mmol) in THF (60 ml). The reaction mixture was stirred at 0° C. for 1 h. The reaction mixture was filtered through a pad of celite, NaBH ₄ (0.705 g, 18.64 mmol) was added to the filtrate and stirred for 2 h. The reaction mixture was diluted with saturated aqueous Na ₂ CO ₃ and stirred for 10 min, then extracted with ethyl acetate. The combined organic layers were washed with saturated sodium chloride solution, dried (Na ₂ SO ₄ ), filtered and the solvent was evaporated. Purification by flash column chromatography (eluting with a 40-50% EtOAc-Heptane gradient) gave the title compound (0.734 g, 52.6%) as a crystalline solid.

¹ H NMR (DMSO-d6, 500 MHz) δ [ppm] = 7.85 (s, 1H), 5.50 (t, J=5.8 Hz, 1H), 4.63 (dd, J=5.8, 0.8 Hz, 2H), 4.37 (q, J=7.1 Hz, 2H), 1.33 (d, J=7.1 Hz, 3H).

HPLC-MS (ESI+): [m/z]: 187.90 [M+H] ⁺ .

Ethyl 4-{[(benzyloxy)methoxy]methyl}-1,3-thiazole-2-carboxylate (218)

NaH (60%, 0.106 g, 2.66 mmol) was added to an ice-cold (0°C) solution of ethyl 4-(hydroxymethyl)-1,3thiazole-2-carboxylate (217) (0.415 g, 2.22 mmol) in THF (20 ml) and the reaction mixture was stirred at 0°C for 30 min. [(Chloromethoxy)methyl]benzene (0.416 g, 2.66 mmol) was added and the reaction mixture was allowed to warm to room temperature over 3 hours. The reaction was quenched by adding saturated aqueous NH ₄ Cl and extracted with ethyl acetate. The combined organic layers were combined, washed with saturated sodium chloride solution, dried (Na ₂ SO ₄ ), filtered and the solvent was evaporated. Purification by flash column chromatography (eluting with 30-50% EtOAcHeptane) gave the title compound (0.392 g, 48.9%) as a colorless oil.

1H NMR (CDCl ₃ , 500 MHz) δ [ppm] = 7.52-7.50 (m, 1H), 7.36-7.33 (m, 5H), 4.90 (s, 2H ), 4.86 (d, J=0.8 Hz, 2H), 4.66 (s, 2H), 4.48 (q, J=7.1 Hz, 2H), 1.44 (t, J =7.1, 3H).

HPLC-MS (ESI+): [m/z]: 307.95 [M+H] ⁺ .

4-{[(Benzyloxy)methoxy]methyl}-1,3-thiazole-2-carboxylic acid (219)

- 167 039916

Similar to general procedure 5, LiOH (64 mg, 1.53 mmol) was added to a solution of ethyl 4{[(benzyloxy)methoxy]methyl}-1,3-thiazole-2-carboxylate (218) (85%, 0.39 g , 1.28 mmol) in THF (5 ml),

MeOH (5 ml) and water (5 ml) at room temperature to give the title compound (0.35 g, 88.4%) as a colorless oil after 4 hours.

¹ H NMR (DMSO-d ₆ , 500 MHz) δ [ppm] = 7.95 (s, 1H), 7.36-7.33 (m, 5H), 4.84 (s, 2H) , 4.74 (s, 2H), 4.60 (s, 2H), 4.49 (s, 1H).

HPLC-MS (ESI+): [m/z]: 279.95 [M+H] ⁺ .

4-{[(Benzyloxy)methoxy]methyl}-N-[(3-fluoropyridin-2-yl)methyl]-1,3-thiazole-2-carboxamide (220) o

Similar to general procedure 6, 4-{[(benzyloxy)methoxy]methyl}-1,3-thiazol-2-carboxylic acid (219) (0.350 g, 1.25 mmol), (3-fluoropyridin-2-yl)methanamine dihydrochloride (A2) (0.374 g, 1.88 mmol), DIPEA (0.89 ml, 5.01 mmol) and HATU (0.714 g, 1.88 mmol) in DMF (10 ml) at room temperature for 16 h gave the title compound (0.381 g, 76.1%) as a yellow oil after purification by flash column chromatography (eluting with a 0-5% MeOH-DCM gradient).

1H NMR (CDCl ₃ , 250 MHz) δ [ppm] = 8.42-8.38 (m, 1H), 7.40-7.27 (m, 7H), 4.90 (s, 2H ), 4.86-4.82 (m, 2H), 4.80 (d, J=0.6 Hz, 2H), 4.68 (s, 2H).

HPLC-MS (ESI+): [m/z]: 388.05 [M+H] ⁺ .

N-[(3-Fluoropyridin-2-yl)methyl]-4-(hydroxymethyl)-1,3-thiazole-2-carboxamide (221) o

TFA (1 ml, 13.06 mmol) was added to an ice-cold (0°C) solution of 4-{[(benzyloxy)methoxy]methyl}-N[(3-fluoropyridin-2-yl)methyl]-1,3-thiazole -2-carboxamide (220) (0.185 g, 0.48 mmol) in DCM (4 ml). The reaction mixture was allowed to warm to room temperature and stirred for 16 h. The solvent was evaporated, the resulting residue was dissolved in EtOAc (5 ml) and washed with saturated aqueous NaHCO ₃ solution (20 ml), saturated sodium chloride solution (10 ml), dried (Na ₂ SO ₄ ), filtered and the solvent was evaporated. Purification by flash column chromatography (eluting with a 5% MeOH-DCM gradient) gave the title compound (0.085 g, 73.2%) as a colorless powder.

¹ H NMR (CDCl3, 500 MHz): δ [ppm] = 8.45-8.41 (m, 1H), 8.36 (br s, 1H), 7.45 (s, 1H) , 7.44-7.40 (m, 1H), 7.31-7.26 (m, 1H), 4.85 (dd, 7= 5.1, 1.6 Hz, 2H), 4.83 (d, J=5.6 Hz, 2H), 2.20 (t, J=6.0 Hz, 1H).

HPLC-MS (ESI+): [m/z]: 267.95 [M+H] ⁺ .

N-[(3-Fluoropyridin-2-yl)methyl]-4-formyl-1,3-thiazole-2-carboxamide (222)

IBX (189 mg, 0.70 mmol) was added to a solution of N-[(3-fluoropyridin-2-yl)methyl]-4-(hydroxymethyl)1,3-thiazole-2-carboxamide (221) (60 mg, 0 .22 mmol) in MeCN (5 ml) and the reaction mixture was heated at 80° C. for 3 h. The reaction mixture was cooled to room temperature and filtered through celite. The solution was adsorbed onto silica gel and purified by flash column chromatography (eluting with a 5% MeOH-DCM gradient). The resulting product was further purified by flash column chromatography (eluting with a 30-50% EtOAc-Heptane gradient) to give the title compound (56 mg, 94%) as a colorless solid.

¹ H NMR (CDCl ₃ , 500 MHz): δ [ppm] = 10.10 (s, 1H), 8.46 (br s, 1H), 8.46-8.44 (m, 1H ), 8.37 (s, 1H), 7.46-7.42 (m, 1H), 7.33-7.28 (m, 1H), 4.88 (dd, J=5.1, 1 .7 Hz, 2H).

HPLC-MS (ESI+): [m/z]: 265.95 [M+H] ⁺ .

4-({[2-(1H-1,3-Benzodiazol-2-yl)ethyl]amino}methyl)-N-[(3-fluoropyridin-2-yl)methyl]-1,3-thiazole-2-carboxamide ( illustrative example No. 205)

n

A solution of X-[(3-fluoropyridin-2-yl)methyl]-4-formyl-1,3-thiazol-2-carboxamide (222) (56 mg, 0.21 mmol) in DCE (1 ml) was added to the suspension 2-(1H-1,3-benzodiazol-2-yl)ethan-1-amine dihydrochloride

- 168 039916 (50 mg, 0.21 mmol) and DIPEA (147 µl, 0.84 mmol) in DCE (2 ml). Activated 4A molecular sieves were added and the solution was stirred at room temperature for 2 h, then filtered and the filtrates were evaporated in vacuo. The resulting residue was dissolved in MeOH (10 ml) and cooled to 0°C. NaBH ₄ (12 mg, 0.32 mmol) was added, the mixture was warmed to room temperature and stirred for 1 h, then quenched with saturated Na ₂ CO ₃ solution and extracted with ethyl acetate (2x10 ml). The combined organic extracts were dried (Na ₂ SO ₄ ), filtered and evaporated in vacuo. Purification by basic prep-HPLC gave the title compound (44 mg, 51%) as a colorless foam.

1H NMR (Methanol-b ₄ , 500 MHz): δ [ppm] = 8.31 (dt, J=4.5, 1.2 Hz, 1H), 7.62 (s, 1H), 7 .57 (ddd, J=9.8, 8.5, 1.2 Hz, 1H), 7.46 (dt, J=6.6, 3.3 Hz, 2H), 7.36 (dt, J = 8.5, 4.5 Hz, 1H), 7.18-7.14 (m, 2H), 4.75 (d, J=1.6 Hz, 2H), 3.96 (s, 2H) , 3.13-3.07 (m, 4H).

HPLC-MS (Method C): [m/z]: 411.1 [M+H] ⁺ .

The above general scheme 12:

general technique 13:

methyl 2-(3-{[(tert-butoxy)carbonyl]amino}propanamido)-3-hydroxypropanoate (223)

TEA (8.38 ml, 60.1 mmol) was added to a chilled (0°C) solution of methyl serinate hydrochloride (1:1) (8.5 g, 54.6 mmol) in DCM (250 ml) under nitrogen atmosphere. N-(tert-butoxycarbonyl)-beta-alanine (10.3 g, 54.6 mmol) and DCC (12.4 g, 60.1 mmol) were added in portions. The reaction mixture was allowed to warm to room temperature and stirred for 19 hours. The solvent was evaporated and EtOAc (250 ml) was added. The resulting mixture was heated to 50°C and stirred for 20 min, cooled to room temperature and filtered. The filtrate was evaporated to give a waxy off-white solid (21.7 g). The resulting solid was dissolved in MeOH (200 ml) and silica gel (300 ml) was added. The solvent was evaporated under reduced pressure to give a compound which was dry-loaded onto silica gel. Purification by flash column chromatography (gradient elution with 20% EtOAc/heptane (2 L), 40% EtOAc/heptane (4 L), 80% EtOAc/heptane (2 L), then EtOAc (8 L) gave the title compound (10.4 g, 64.4%) as an off-white solid.

¹ H NMR (CDCl3, 250 MHz): δ [ppm] = 6.62 (s, 1H), 5.12 (s, 1H), 4.65 (dt, J=7.1, 3, 4 Hz, 1H), 3.96 (t, J=3.2 Hz, 2H), 3.80 (s, 3H), 3.55-3.33 (m, 2H), 2.95 (s, 1H), 2.48 (td, J=5.8, 2.6 Hz, 2H), 1.43 (s, 9H).

HPLC-MS (method A): [m/z]: 291 [m+H] ⁺ .

Methyl 2-(2-{[(tert-butoxy)carbonyl]amino}acetamido)-3-hydroxypropanoate (224)

Similar to general procedure 13, methyl serinate hydrochloride (3 g, 19.28 mmol), TEA (2.96 ml, 21.21 mmol), N-(t-butoxycarbonyl)glycine (3.38 g, 19.28 mmol) and DCC (4.38 g, 21.21 mmol) in DCM (100 mL) gave the title compound (4.5 g, 69%) as a pale yellow oil after purification by flash column chromatography eluting with a gradient of 20 -100% EtOAc/heptane.

1H NMR (CDCl3, 250 MHz): δ [ppm] = 7.07 (d, J=7.4 Hz, 1H), 5.32 (s, 1H), 4.77-4.60 ( m, 1H), 3.99 (m, 2H), 3.87 (m, 2H), 3.81 (s, 3H), 3.04 (s, 1H), 1.48 (s, 9H).

HPLC-MS (Method A): [m/z]: 298.95 [M+Na] ⁺ .

General Procedure 14: Methyl 2-(2-{[(tert-butoxy)carbonyl]amino}ethyl)-4,5-dihydro-1,3-oxazole-4carboxylate(225)

Methyl 2-(3-{[(tert-butoxy)carbonyl]amino}propanamido)-3-hydroxypropanoate (223) (98%, 10.42 g, 35.17 mmol) was dissolved in anhydrous DCM (280 mL) under atmospheric nitrogen. The reaction mixture was cooled in a CO ₂ /MeCN bath (about -50° C.) and stirred for 30 minutes. DAST (5.58 ml, 42.21 mmol) was added dropwise and the resulting mixture was stirred at -50°C for 2.25 hours. K ₂ CO ₃ (4.86 g, 35.17 mmol) was added to one reception, and the reaction mixture was left to stir for 20 min before warming to room temperature. The reaction mixture was then placed in a water bath and water (60 ml) was carefully added (gas evolution observed) to the reaction mixture, then 2M NaOH solution (5 ml) was added. The reaction mixture was stirred for another 10 min and the layers were separated. The aqueous layer was extracted with dichloromethane (2x50 ml). The combined organic layers were dried (MgSO ₄ ), filtered and evaporated to give the title compound (10.3 g) as an orange oil. The crude product was used in the next step without further purification.

- 169 039916

Ή NMR (CDCl3, 500 MHz): δ [ppm] = 5.14 (s, 1H), 4.72 (ddt, J=10.3, 7.6, 1.2 Hz, 1H), 4.47 (dd, J=8.7,

7.7 Hz, 1H), 4.39 (dd, J=10.6, 8.8 Hz, 1H), 3.78 (s, 3H), 3.49-3.36 (m, 2H), 2.48 (td, J=6.2, 1.2 Hz, 2H), 1.42 (s,

9H).

HPLC-MS (method A): [m/z]: 273 [M+H] ⁺ .

Methyl 2-({[(tert-butoxy)carbonyl]amino}methyl)-4,5-dihydro-1,3-oxazole-4-carboxylate (226)

Similar to general procedure 14, methyl 2-(2-{[(tert-butoxy)carbonyl]amino}acetamido)-3hydroxypropanoate (224) (2.5 g, 7.42 mmol) and DAST (1.18 ml, 8. 9 mmol) in DCM (70 mL) gave the title compound (2.02 g, quant) as a yellow oil.

Ή NMR (CDCl ₃ , 250 MHz): δ [ppm] = 5.12 (s, 1H), 4.86-4.68 (m, 1H), 4.62-4.42 (m, 2H), 4.11-3.89 (m, 2H), 3.82 (s, 3H), 1.44 (s, 9H).

HPLC-MS (method A): [m/z]: 258.95 [M+H] ⁺ .

General Procedure 15: Methyl 2-(2-{[(tert-butoxy)carbonyl]amino}ethyl)-1,3-oxazole-4-carboxylate (227)

Bromo(trichloro)methane (10.4 mL, 105.56 mmol) was added to a solution of methyl 2-(2-{[(tert-butoxy)carbonyl]amino}ethyl)-4,5-dihydro-1,3-oxazole-4 -carboxylate (225) (93.2%, 10.28 g, 35.19 mmol) in anhydrous DCM (300 ml) under nitrogen, cooling to 0°C. DBU (15.75 mL, 105.56 mmol) was added dropwise and the resulting mixture was allowed to warm to room temperature and stirred for 2.5 hours. Saturated citric acid aqueous solution (25 mL) was added followed by water (100 mL). The reaction mixture was intensively stirred for 10 min. The layers were separated and the aqueous layer was extracted with dichloromethane (3x100 ml). The organic layer was dried (MgSO ₄ ), filtered and evaporated to give a dark brown oil (14.6 g). The resulting crude oil was applied to silica gel with MeOH (200 ml). Filter through a pad of silica gel [gradient elution with heptane (2x500 ml), 25% EtOAc/heptane (2x500 ml), and 50% EtOAc/heptane (11x500 ml)] to give the title compound (8.63 g) as yellow oil that hardens on standing.

Ή NMR (CDCl ₃ , 500 MHz): δ [ppm] = 8.16 (s, 1H), 4.99 (s, 1H), 3.90 (s, 3H), 3.57 (q , J=6.0 Hz, 2H), 2.99 (t, J=6.3 Hz, 2H), 1.40 (s, 9H).

HPLC-MS (method A): [m/z]: 293.0 [M+Na] ⁺ .

Methyl 2-({[(tert-butoxy)carbonyl]amino}methyl)-4,5-dihydro-1,3-oxazole-4-carboxylate (228)

Similar to general procedure 15, methyl 2-({[(tert-butoxy)carbonyl]amino}methyl)-4,5-dihydro-1,3oxazole-4-carboxylate (226) (2.02 g, 7.43 mmol) , DBU (3.33 ml, 22.29 mmol) and bromo(trichloro)methane (2.2 ml, 22.29 mmol) in DCM (75 ml) gave the title compound (1.04 g, 55%) as a colorless oil after purification by flash column chromatography eluting with a 0-100% TBME/heptane gradient.

Ή NMR (CDCl ₃ , 500 MHz): δ [ppm] = 8.19 (s, 1H), 5.19 (broad s, 1H), 4.49 (d, J=5.5 Hz , 2H), 3.92 (s, 3H), 1.45 (s, 9H).

HPLC-MS (Method A): [m/z]: 278.95 [M+Na]+.

2-(2-{[(tert-Butoxy)carbonyl]amino}ethyl)-1,3-oxazole-4-carboxylic acid (229)

Similarly, using General Procedure 5, LiOH (1.02 g, 42.76 mmol) and methyl 2-(2{[(tert-butoxy)carbonyl]amino}ethyl)-1,3-oxazole-4-carboxylate ( 227) (97%, 6.62 g, 23.76 mmol) in THF (100 mL) and water (25 mL) gave the title compound (4.8 g, 79%) as an off-white powder.

Ή NMR (DMSO-d6, 500 MHz): δ [ppm] = 8.62 (s, 1H), 6.94 (t, J=5.4 Hz, 1H), 3.31-3, 26 (m, 2H, partial overlap with HOD signals), 2.87 (t, J=6.8 Hz, 2H), 1.35 (s, 9H).

HPLC-MS (method A): [m/z]: 278.9 [M+Na] ⁺ .

2-({[(tert-Butoxy)carbonyl]amino}methyl)-1,3-oxazole-4-carboxylic acid (230)

- 170 039916

Similar to general procedure 5, methyl 2-({[(tert-butoxy)carbonyl]amino}methyl)-4,5-dihydro-1,3oxazole-4-carboxylate (228) (1.04 g, 4.06 mmol) and LiOH (0.15 g, 6.09 mmol) in THF/water (20 mL/5 mL) gave the title compound (1.07 g, quant) as an off-white solid.

1H NMR (CDCl ₃ , 500 MHz): δ [ppm] = 8.29 (s, 1H), 5.39 (broad s, 1H), 4.54 (d, J=4.9 Hz , 2H), 1.47 (s, 9H).

HPLC-MS (method A): [m/z]: 265 [M+Na] ⁺ .

tert-Butyl N-[2-(4-{[(3-fluoropyridin-2-yl)methyl]carbamoyl}-1,3-oxazol-2-yl)ethyl]carbamate (231)

Similarly, using the general procedure 6, 2-(2-{[(tert-butoxy)carbonyl]amino}ethyl)1,3-oxazole-4-carboxylic acid (229) (3.78 g, 14.75 mmol) , HATU (6.17 g, 16.23 mmol), (DIPEA) (8.48 ml, 48.68 mmol), (3-fluoropyridin-2-yl)methanamine dihydrochloride (A2) (2.94 g, 14 .75 mmol) in THF/water (100 mL/40 mL) gave the title compound (4.83 g, 87%) as a yellow glassy solid after purification by flash column chromatography (eluting with a gradient of 0-100 % EtOAc/heptane).

1H NMR (DMSO-06, 500 MHz): δ [ppm] = 8.51 (s, 2H), 8.40 (d, J=4.4 Hz, 1H), 7.70 (t, J=9.3Hz, 1H), 7.45-7.37(m, 1H), 6.98(t, J=5.5Hz, 1H), 4.62(d, J=5.5 Hz, 2H), 3.32-3.28 (m, 2H, partial overlap with HOD signals), 2.90 (t, J=6.7 Hz, 2H), 1.35 (s, 9H).

HPLC-MS (method A): [m/z]: 365.0 [M+H] ⁺ .

tert-Butyl N-(2-{4-[(pyridin-2-ylmethyl)carbamoyl]-1,3-oxazol-2-yl}ethyl)carbamate (232)

Similarly, using the general procedure 6, 2-(2-{[(tert-butoxy)carbonyl]amino}ethyl)1,3-oxazole-4-carboxylic acid (229) (4 g, 15.61 mmol), DIPEA (5.44 ml, 31.22 mmol), HATU (8.9 g, 23.41 mmol), pyridin-2-ylmethanamine (2.41 ml, 23.41 mmol) in DMF (65 ml) gave the indicated title compound (6.03 g) as a yellow viscous oil after purification by flash column chromatography (eluting with a 0-100% EtOAc/heptane gradient).

¹ H NMR (DMSO-d ₆ , 250 MHz): δ [ppm] = 8.74 (t, J=5.9 Hz, 1H), 8.54-8.48 (m, 2H), 7.75 (td, J=7.7, 1.8 Hz, 1H), 7.32-7.22 (m, 2H), 6.98 (s, 1H), 4.52 (d, J= 6.0 Hz, 2H), 3.29 (d, J=6.5 Hz, 2H), 2.90 (t, J=6.7 Hz, 2H), 1.34 (s, 9H).

HPLC-MS (method A): [m/z]: 347.1 [M+H]+.

tert-Butyl N-[2-(4-{[(3-chloropyridin-2-yl)methyl]carbamoyl}-1,3-oxazol-2-yl)ethyl]carbamate (233)

Similar to general procedure 6, 2-(2-{[(tert-butoxy)carbonyl]amino}ethyl)-1,3-oxazole-4carboxylic acid (229) (295 mg, 0.76 mmol, 66%), (3 -chloropyridin-2-yl)methanamine dihydrochloride (246 mg, 1.14 mmol), TEA (0.1 ml, 0.76 mmol) and HATU (0.43 g, 1.14 mmol) in DMF (10 ml) gave the title compound (202 mg, 59, 85% pure) as a yellow oil after purification by flash column chromatography (eluting with a 0-10% MeOH/DCM gradient).

1H NMR (DMSO-d ₆ , 250 MHz): δ [ppm] = 8.58-8.45 (m, 2H), 7.96 (dd, J=8.1, 1.4 Hz, 1H), 7.40 (dd, J=8.1, 4.7 Hz, 1H), 7.00 (t, J=6.2 Hz, 1H), 4.65 (d, J=5.4 Hz, 2H), 2.97-2.86 (m, 4H), 1.35 (s, 9H).

HPLC-MS (method A): [m/z]: 381.05 [M+H] ⁺ .

tert-Butyl N-[(4-{[(3-fluoropyridin-2-yl)methyl]carbamoyl}-1,3-oxazol-2-yl)methyl]karbamate (234)

Similar to general procedure 6, 2-({[(tert-butoxy)carbonyl]amino}methyl)-1,3-oxazole-4carboxylic acid (230) (0.5 g, 2.06 mmol), (3-fluoropyridine- 2-yl)methanamine dihydrochloride (A2) (0.41 g, 2.06 mmol), DIPEA (1.19 ml, 6.81 mmol) and HATU (0.86 g, 2.27 mmol) in THF (15 mL) and DMF (6 mL) gave the title compound (0.744 g, 98%) as a colorless oil after purification by flash column chromatography eluting with a 0-100% EtOAc/heptane gradient.

1H NMR (DMSO-d ₆ , 500 MHz): δ [ppm] = 8.56 (s, 1H), 8.51 (t, J=5.2 Hz, 1H), 8.38 (d , J=4.6 Hz, 1H), 7.73-7.65 (m, 1H), 7.55 (t, J=5.8 Hz, 1H), 7.44-7.37 (m, 1H), 4.65-4.59 (m, 2H), 4.29 (d, J=5.9 Hz, 2H), 1.39 (s, 9H).

- 171 039916

HPLC-MS (method A): [m/z]: 351.0 [M+H]+.

2-(2-Aminoethyl)-N-[(3-fluoropyridin-2-yl)methyl]-1,3-oxazole-4-carboxamide dihydrochloride (235) ^nh ₂

2HCI

Similarly, using General Procedure 2, tert-butyl X-[2-(4-{[(3-fluoropyridin-2yl)methyl]carbamoyl}-1,3-oxazol-2-yl)ethyl]carbamate (231) ( 4.83 g, 12.86 mmol), a 4M solution of HC in 1,4dioxane (50 mL) and DCM (80 mL) gave the title compound (4.61 g) as an off-white solid.

¹ H NMR (DMSO-66, 500 MHz): δ [ppm] = 8.71 (t, J=5.8 Hz, 1H), 8.59 (s, 1H), 8.42-8 .36 (m, 1H), 8.13 (brs, 3H), 7.76-7.68 (m, 1H), 7.46-7.39 (m, 1H), 4.64 (dd , J=5.8, 1.4 Hz, 2H), 3.25 (pseudo square, J=6.1 Hz, 2H), 3.20-3.14 (m, 2H).

HPLC-MS (method F): [m/z]: 265.1 [M+H] ⁺ .

2-(2-Aminoethyl)-N-(pyridin-2-ylmethyl)-1,3-oxazole-4-carboxamide dihydrochloride (236)

Similarly, using General Procedure 2, tert-butyl X-(2-{4-[(pyridin-2ylmethyl)carbamoyl]-1,3-oxazol-2-yl}ethyl)carbamate (232) (90%, 6, 03 g, 15.68 mmol), 12M HC (26.13 ml) in MeOH gave the title compound (5.78 g) as a creamy foam.

1H NMR (Methanol-b ₄ , 250 MHz): δ [ppm] = 8.80 (dd, J=5.9, 0.8 Hz, 1H), 8.62 (td, J=8, 0, 1.6Hz, 1H), 8.44(s, 1H), 8.10(d, J=7.8Hz, 1H), 8.01(t, J=6.3Hz, 1H) , 4.93 (s, 2H), 3.51-3.43 (m, 2H), 3.26 (t, J=6.4 Hz, 2H).

HPLC-MS (method A): [m/z]: 246.9 [M+H] ⁺ .

2-(2-Aminoethyl)-N-[(3-chloropyridin-2-yl)methyl]-1,3-oxazole-4-carboxamide dihydrochloride (237)

Similarly, using General Procedure 2, tert-butyl X-[2-(4-{[(3-chloropyridin-2yl)methyl]carbamoyl}-1,3-oxazol-2-yl)ethyl]carbamate (233) ( 85%, 202 mg, 0.45 mmol), MeOH (5 ml) and 4M HC in dioxane (1.65 ml) gave the title compound (125 mg, 62%) as a brown solid.

1H NMR (DMSO-66, 500 MHz): δ [ppm] = 8.67 (t, J=5.6 Hz, 1H), 8.61 (s, 1H), 8.52 (dd, J=4.7, 1.4Hz, 1H), 8.15(s, 3H), 7.96(dd, J=8.1, 1.4Hz, 1H), 7.40(dd, J = 8.1, 4.7 Hz, 1H), 4.67 (d, J=5.6 Hz, 2H), 3.29-3.23 (m, 2H), 3.21-3.16 ( m, 2H).

HPLC-MS (method A): [m/z]: 281.1 [M+H] ⁺ .

2-(Aminomethyl)-X-[(3-fluoropyridin-2-yl)methyl]-1,3-oxazole-4-carboxamide dihydrochloride (238)

Similar to general procedure 2, t-butyl X-[(4-{[(3-fluoropyridin-2-yl)methyl]carbamoyl}-1,3oxazol-2-yl)methyl]carbamate (234) (744 mg, 2, 02 mmol) and a 4M solution of HC in 1,4-dioxane (15 ml) in DCM (20 ml) gave the title compound (668 mg, quant.) as a white solid.

¹ H NMR (DMSO-66, 500 MHz): δ [ppm] = 8.79 (s, 1H), 8.73 (br s, 3H), 8.54 (t, J=5, 6 Hz, 1H), 8.39 (d, J=4.7 Hz, 1H), 7.80-7.68 (m, 1H), 7.49-7.38 (m, 1H), 4, 66 (d, J=4.6 Hz, 2H), 4.32 (t, J=5.5 Hz, 2H).

HPLC-MS (method F): [m/z]: 251.1 [M+H] ⁺ .

2-(2-{[2-(1H-1,3-Benzodiazol-2-yl)ethyl]amino}ethyl)-X-[(3-fluoropyridin-2-yl)methyl]-1,3-oxazol- 4carboxamide trihydrochloride (illustrative example No. 127)

Similar to general procedure 8, 2-(2-Aminoethyl)-X-[(3-fluoropyridin-2-yl)methyl]-1,3-oxazole-4-carboxamide dihydrochloride (235) (1 g, 2.97 mmol), K -(2-nitrophenyl)prop-2-enamide (D) (0.63 g, 3.26

- 172 039916 mmol) and DBU (1.5 ml, 10.08 mmol) in MeCN (30 ml) at room temperature overnight gave a crude intermediate product which was reacted with iron powder (0.5 g) in AcOH (10 ml) under nitrogen atmosphere at 80° C. for 3 hours. The resulting crude product was purified by basic prep-HPLC followed by purification by flash column chromatography (eluting with a gradient of 0-30% MeOH in DCM) to give 2-(2- {[2-(1H-1,3-benzodiazol-2-yl)ethyl]amino}ethyl)-N-[(3-fluoropyridin-2yl)methyl]-1,3-oxazol-4-carboxamide. The resulting free base was dissolved in MeOH (10 ml) and 12M hydrogen chloride solution (1.5 ml) was added. The solvent was then evaporated in vacuo to give the title compound (333 mg, 22%) as a white solid.

1H NMR (D2O, 500 MHz): δ [ppm] = 8.34-8.30 (m, 2H), 7.86 (ddd, J=9.6, 8.7, 1.2 Hz , 1H), 7.74-7.69 (m, 2H), 7.62-7.57 (m, 1H), 7.57-7.52 (m, 2H), 3.74-3.66 (m, J=4.3 Hz, 4H), 3.64 (t, J=6.7 Hz, 2H), 3.33 (t, J=6.7 Hz, 2H).

HPLC-MS (method C): [m/z]: 409.1 [M+H] ⁺ .

2-(2-{bis[2-(1H-1,3-Benzodiazol-2-yl)ethyl]amino}ethyl)-X-[(3-fluoropyridin-2-yl)methyl]-1,3oxazol-4 -carboxamide tetrahydrochloride (illustrative example No. 128)

Similar to General Procedure 7, 2-(2-Aminoethyl)-X-[(3-fluoropyridin-2-yl)methyl]-1,3-oxazole-4-carboxamide dihydrochloride (Illustrative Example No. 127) (305 mg, 0.9 mmol ), 2-(2-chloroethyl)-III1,3-benzodiazol hydrochloride (216 mg, 1 mmol) and DIPEA (3.15 ml, 18 mmol) in DMF (5 ml) gave the title compound in free base form after purification by basic prepHPLC. The resulting free base was treated with 12M HC (0.5 ml) in MeOH (5 ml) to give the title compound as tetrahydrochloride (5 mg, 0.8%) as a brown residue.

1H NMR (Methanol ₄ , 500 MHz): δ [ppm] = 8.56-8.52 (m, 1H), 8.45 (s, 1H), 8.21-8.15 ( m, 1H), 7.90-7.84 (m, 2H), 7.84-7.77 (m, 3H), 7.69-7.60 (m, 3H), 7.58 (ddd, J=8.2, 7.4, 0.6 Hz, 1H), 5.32-5.22(m, 2H), 4.90(s, 2H), 4.17-4.11(m, 2H), 4.02-3.94 (m, 2H), 3.93-3.86 (m, 2H), 3.78 (t, J=6.3 Hz, 2H), 3.48 (t , J=6.3 Hz, 2H).

HPLC-MS (method C): [m/z]: 553.2 [M+H] ⁺ .

2-(2-{[2-(1H-1,3-Benzodiazol-2-yl)ethyl]amino}ethyl)-X-(pyridin-2-ylmethyl)-1,3-oxazole-4carboxamide trihydrochloride (illustrative example No. 171)

Similarly, using the general procedure 8, 2-(2-Aminoethyl)-X-(pyridin-2-ylmethyl)-1,3oxazol-4-carboxamide dihydrochloride (236) (86%, 1.2 g, 3.25 mmol ), DBU (1.46 ml, 9.76 mmol) and N-(2nitrophenyl)prop-2-enamide (D) (0.69 g, 3.58 mmol) in MeCN (30 ml) gave the crude intermediate, which reacted with iron powder (0.52 g) in AcOH (10 ml) to give the title compound (0.22 g, 12%) as a pale brown solid.

1H NMR (D2O), 500 MHz: δ [ppm] = 8.72 (d, J=5.8 Hz, 1H), 8.54 (td, J=8.0, 1.5 Hz , 1H), 8.46 (s, 1H), 8.00-7.94 (m, 2H), 7.81 (dt, J=6.2, 3.2 Hz, 2H), 7.63 ( dt, J=6.2, 3.3 Hz, 2H), 4.88 (s, 2H), 3.83-3.70 (m, 6H), 3.43 (t, J=6.9 Hz , 2H).

HPLC-MS (method C): [m/z]: 391.2 [M+H]+.

2-(2-{[2-(1H-1,3-Benzodiazol-2-yl)ethyl]amino}ethyl)-N-[(3-chloropyridin-2-yl)methyl]-1,3-oxazol- 4carboxamide (illustrative example No. 173)

Similarly, using the general procedure 8, 2-(2-Aminoethyl)-X-[(3-chloropyridin-2yl)methyl]-1,3-oxazole-4-carboxamide dihydrochloride (237) (75%, 120 mg, 0 .25 mmol), DBU (76 μl, 0.51 mmol) and N-(2-nitrophenyl)prop-2-enamide (D) (48 mg, 0.25 mmol) in MeCN (5 ml) gave crude intermediate 173 039916 the exact product which was reacted with iron powder (36.9 mg) in AcOH (2 ml) to give the title compound (4.6 mg, 3%) as a clear film after purification by basic pre-HPLC.

1H NMR (Methanol-d ₄ , 500 MHz): δ [ppm] = 8.44 (dd, J=4.7, 1.4 Hz, 1H), 8.28 (s, 1H), 7 .86 (dd, J=8.1, 1.4 Hz, 1H), 7.47-7.41 (m, 2H), 7.33 (dd, J=8.1, 4.8 Hz, 1H ), 7.19-7.14 (m, 2H), 4.70 (s, 2H), 3.18-3.04 (m, 8H).

HPLC-MS (method C): [m/z]: 425.2 [M+H] ⁺ .

2-({[2-(1H-1,3-Benzodiazol-2-yl)ethyl]amino}methyl)-N-[(3-fluoropyridin-2-yl)methyl]-1,3-oxazol-4carboxamide ( illustrative example No. 206)

Similar to general procedure 8, 2-(aminomethyl)-N-[(3-fluoropyridin-2-yl)methyl]-1,3-oxazole-4-carboxamide dihydrochloride (238) (300 mg, 0.93 mmol), N-( 2-nitrophenyl)prop-2-enamide (D) (196 mg, 1.02 mmol) and DBU (471.1 μl, 3.16 mmol) in MeCN (10 ml) gave a crude intermediate which was reacted with iron powder (156 mg, 2.79 mmol) in AcOH (5 mL) to give the title compound (20 mg, 27%) as a white solid after purification by basic prep-HPLC, followed by flash column chromatography eluting with gradient 2-20% MeOH/DCM.

1H NMR (DMSO-a ₆ , 500 MHz): δ [ppm] = 12.15 (s, 1H), 8.57 (s, 1H), 8.53 (t, J=5.6 Hz , 1H), 8.38 (dt, J=4.6, 1.3 Hz, 1H), 7.70 (m, 1H), 7.51 (d, J=6.9 Hz, 1H), 7 .41 (m, 2H), 7.15-7.07 (m, 2H), 4.62 (dd, J=5.6, 1.3 Hz, 2H), 3.92 (s, 2H), 3.04-2.99 (m, 2H), 2.98-2.94 (m, 2H).

HPLC-MS (Method C): [m/z]: 395.1 [M+H] ⁺ .

2-{2-[(1H-1,3-Benzodiazol-2-ylmethyl)amino]ethyl}-H-[(3-fluoropyridin-2-yl)methyl]-1,3-oxazole-4carboxamide trihydrochloride (illustrative example No. 126)

Similarly, using the general procedure 3, 2-(2-aminoethyl)-N-[(3-fluoropyridin-2yl)methyl]-1,3-oxazole-4-carboxamide dihydrochloride (235) (100 mg, 0.3 mmol ), 1H-benzimidazole-2carbaldehyde (43.3 mg, 0.3 mmol), DIPEA (0.207 ml, 1.19 mmol) and anhydrous MgSO ₄ (200 mg) in DCM (10 ml) at room temperature for 16 h , followed by the addition of NaBH ₄ (17 mg, 0.45 mmol) gave the compound as the free base after purification by basic prep-HPLC. The resulting residue was redissolved in MeOH (5 ml) and treated with 12M HC (1 ml) for 30 min to give the title compound (64 mg, 42%) as a white hygroscopic solid.

¹ H NMR (Methanol-d ₄ , 500 MHz): δ [ppm] = 8.57 (dd, J=5.4, 0.9 Hz, 1H), 8.46 (s, 1H), 8.26-8.19 (m, 1H), 7.88 (ddt, J=13.4, 8.9, 4.1 Hz, 3H), 7.68 (td, J=6.3.5 .5, 2.1 Hz, 2H), 5.02 (s, 2H), 4.94 (s, 2H), 3.83 (t, J=6.3 Hz, 2H), 3.47 (t , J=6.3 Hz, 2H).

HPLC-MS (method C): [m/z]: 395.2 [M+H]+.

2-{2-[(1H-1,3-Benzodiazol-2-ylmethyl)amino]ethyl}-N-(pyridin-2-ylmethyl)-1,3-oxazol-4carboxamide (illustrative example No. 137)

Similarly, using the general procedure 3, 2-(2-aminoethyl)-N-(pyridin-2-ylmethyl)-1,3oxazol-4-carboxamide dihydrochloride (236) (430 mg, 1.31 mmol), 1H-benzimidazole -2-carbaldehyde (229 mg, 1.57 mmol) and DIPEA (0.91 ml, 5.23 mmol) in MeOH (20 ml) at room temperature for 17 h, followed by the addition of NaBH ₄ (74 mg, 1 .96 mmol) gave the title compound (380 mg, 77%) as a yellow solid after purification by flash column chromatography eluting with a 0-20% MeOH/DCM gradient.

1H NMR (DMSO-d ₆ , 500 MHz): δ [ppm] = 12.17 (s, 1H), 8.72 (t, J=5.9 Hz, 1H), 8.53-8 .46 (m, 2H), 7.74 (td, J=7.7, 1.7 Hz, 1H), 7.48 (m, 2H), 7.31-7.22 (m, 2H), 7.12 (m, 2H), 4.52 (d, J=6.0 Hz, 2H), 3.95 (s, 2H), 3.032.93 (m, 4H).

HPLC-MS (method C): [m/z]: 377.2 [M+H] ⁺ .

2-{2-[(1H-1,3-Benzodiazol-2-ylmethyl)amino]ethyl}-N-[(3-chloropyridin-2-yl)methyl]-1,3-oxazole-4carboxamide (illustrative example no. 141)

- 174 039916

Similarly, using the general procedure 3, 2-(2-Aminoethyl)-N-[(3-chloro-pyridin-2yl)methyl]-1,3-oxazole-4-carboxamide dihydrochloride (237) (118 mg, 0.31 mmol ), 1H-benzimidazole-2carbaldehyde (54 mg, 0.37 mmol) and DIPEA (0.216 ml, 1.24 mmol) in MeOH (4 ml) at room temperature for 24 h, followed by the addition of NaBH ₄ (18 mg, 0.46 mmol) gave the title compound (51 mg, 40%) as a brown solid after purification by basic pre-HPLC.

1H NMR (Methanol-d ₄ , 500 MHz): δ [ppm] = 8.41 (dd, J=4.8, 1.4 Hz, 1H), 8.29 (s, 1H), 7 .86 (dd, J=8.1, 1.4 Hz, 1H), 7.53-7.49 (m, 2H), 7.34-7.30 (m, 1H), 7.21-7 .18 (m, 2H), 4.76 (s, 2H), 4.08 (s, 2H), 3.14-3.09 (m, 2H), 3.07-3.03 (m, 2H ).

HPLC-MS (Method D): [m/z]: 411.2 [M+H]+.

2-(2-{[2-(4,5-Difluoro-1H-1,3-benzodiazol-2-yl)ethyl]amino}ethyl)-N-[(3-fluoro-pyridin-2-yl)methyl]1 ,3-oxazole-4-carboxamide (illustrative example No. 257)

Similar to general procedure 8, 2-(2-Aminoethyl)-N-[(3-fluoropyridin-2-yl)methyl]-1,3-oxazole-4-carboxamide dihydrochloride (235) (350 mg, 1.04 mmol), N -(2,3-difluoro-6-nitrophenyl)prop-2-enamide (K10) (250 mg, 1.10 mmol) and DBU (465 µl, 3.11 mmol) in MeCN (10 ml) gave the intermediate (193 mg) after purification by flash column chromatography (elution with a 0-20% MeOH/DCM gradient). The resulting intermediate was further reacted with iron powder (88 mg, 1.57 mmol) in AcOH (2 mL) to give the title compound (14 mg, 8%) as a pale yellow solid after purification by flash column chromatography ( 0-30% MeOH/DCM gradient elution) followed by basic prep-HPLC.

1H NMR (DMSO-66, 500 MHz): δ [ppm] = 8.55-8.47 (m, 2H), 8.38 (d, J=4.6 Hz, 1H), 7, 74-7.67 (m, 1H), 7.44-7.36 (m, 1H), 7.23 (dd, J=8.8, 3.6 Hz, 1H), 7.17-7, 09 (m, 1H), 4.61 (d, J=4.8 Hz, 2H), 3.02-2.91 (m, 8H).

HPLC-MS (method d): [m/z]: 445.2 [M+H] ⁺ .

2-(2-{[2-(4-Chloro-1H-1,3-benzodiazol-2-yl)ethyl]amino}ethyl)-N-[(3-fluoropyridin-2-yl)methyl]-1, 3oxazole-4-carboxamide (illustrative example No. 226) ^C|

HN

V/

Similar to general procedure 8, 2-(2-Aminoethyl)-N-[(3-fluoropyridin-2-yl)methyl]-1,3-oxazole-4-carboxamide dihydrochloride (235) (340 mg, 1.01 mmol), N -(3-chloro-2-nitrophenyl)prop-2-enamide (H) (245 mg, 1.08 mmol) and DBU (147 μl, 0.98 mmol) in MeCN (10 ml) gave a crude intermediate which reacted with iron powder (81 mg, 1.45 mmol) in AcOH (4 mL) to give the title compound (31 mg, 19%) as a white solid after purification by basic prep-HPLC followed by flash column chromatography (elution with a 0-40% MeOH/DCM gradient).

1H NMR (DMSO-d6, 500 MHz): δ [ppm] = 8.51 (m, 2H), 8.38 (dt, J=4.6, 1.2 Hz, 1H), 7, 70 (ddd, J=10.0, 8.3, 1.2 Hz, 1H), 7.41 (m, 2H), 7.19 (dd, J=7.7, 0.8 Hz, 1H) , 7.12 (t, J=7.7 Hz, 1H), 4.62 (dd, J=5.6, 1.3 Hz, 2H), 3.09-2.95 (m, 8H).

HPLC-MS (method C): [m/z]: 443.1 [M+H] ⁺ .

N-[(3-Fluoropyridin-2-yl)methyl]-2-(2-{[2-(4-methoxy-1H-1,3-benzodiazol-2-yl)ethyl]amino}ethyl)1,3 -oxazole-4-carboxamide (illustrative example No. 227)

- 175 039916

Similar to general procedure 8, 2-(2-aminoethyl)-N-[(3-fluoropyridin-2-yl)methyl]-1,3-oxazole-4-carboxamide dihydrochloride (235) (301 mg, 0.89 mmol), N -(2-methoxy-6-nitrophenyl)prop-2-enamide (I) (198 mg, 0.89 mmol) and DBU (0.4 ml, 2.68 mmol) in MeCN (10 ml) gave the crude intermediate , which was partially purified by flash column chromatography eluting with a gradient of 010% MeOH in DCM. The resulting intermediate was further reacted with iron powder (86 mg) in AcOH (3 mL) to give the title compound (2 mg, 1%) as a white solid after purification by basic prep-HPLC, followed by flash column chromatography ( eluting with a gradient of 0-20% MeOH/DCM).

1H NMR (MeOD, 500 MHz): δ [ppm] = 8.30 (d, J=4.7 Hz, 1H), 8.26 (s, 1H), 7.60-7.54 ( m, 1H), 7.39-7.32 (m, 1H), 7.09 (t, J=8.0 Hz, 1H), 7.03 (d, 1H), 6.70 (d, J = 7.8 Hz, 1H), 4.65 (d, J=1.4 Hz, 2H), 3.93 (s, 3H), 3.2-3.16 (m, 4H), 3.14-3.05 (m, 4H).

HPLC-MS (method d): [m/z]: 439.2 [M+H] ⁺ .

2-(2-{[2-(4-Fluoro-1H-1,3-benzodiazol-2-yl)ethyl]amino}ethyl)-N-[(3-fluoropyridin-2-yl)methyl]-1, 3oxazole-4-carboxamide (illustrative example No. 228)

Similar to general procedure 8, 2-(2-Aminoethyl)-N-[(3-fluoropyridin-2-yl)methyl]-1,3-oxazole-4-carboxamide dihydrochloride (235) (200 mg, 0.59 mmol), N -(3-fluoro-2-nitrophenyl)prop-2-enamide (G) (198 mg, 0.89 mmol) and DBU (266 μl, 1.78 mmol) in MeCN (10 ml) gave an intermediate which was partially purified flash column chromatography eluting with a gradient of 0-10% MeOH in DCM. The resulting intermediate was further reacted with iron powder (114 mg) in AcOH (2 mL) to give the title compound (10 mg, 5%) as an off-white solid after purification by basic prep-HPLC, followed by flash column. chromatography (elution gradient 030% MeOH/DCM) and kp-NH flash chromatography (elution gradient 0-4% MeOH/DCM).

1H NMR (DMSO-d ₆ , 500 MHz): δ [ppm] = 8.53-8.45 (m, 2H), 8.39-8.35 (m, 1H), 7.72- 7.67 (m, 1H), 7.427.37 (m, 1H), 7.26 (d, J=8.0 Hz, 1H), 7.11-7.05 (m, 1H), 6.93 -6.88 (m, 1H), 4.61 (d, J=4.4 Hz, 2H), 3.01-2.91 (m, 8H).

HPLC-MS (method d): [m/z]: 427.1 [M+H] ⁺ .

2-(2-{[2-(7-Chloro-4-fluoro-1H-1,3-benzodiazol-2-yl)ethyl]amino}ethyl)-N-[(3-fluoropyridin-2yl)methyl]- 1,3-oxazole-4-carboxamide (illustrative example No. 247)

Similar to general procedure 8, 2-(2-Aminoethyl)-N-[(3-fluoropyridin-2-yl)methyl]-1,3-oxazole-4-carboxamide dihydrochloride (235) (300 mg, 0.89 mmol), N -(6-chloro-3-fluoro-2-nitrophenyl)prop-2-enamide (K2) (272 mg, 0.89 mmol) and DBU (0.398 ml, 2.67 mmol) in MeCN (15 ml) gave crude an intermediate which was purified by flash column chromatography eluting with a gradient of 015% MeOH in DCM. The resulting intermediate was further reacted with iron powder (145 mg) in AcOH (3 mL) to give the title compound (7 mg, 2%) as a beige solid after purification by flash column chromatography (eluting with a 0-35% gradient). MeOH/DCM), followed by kp-NH column chromatography (eluting with a 0-15% MeOH/DCM gradient) and basic prep-HPLC.

¹ H NMR (MeOD, 500 MHz): δ [ppm] = 8.34 (d, J=4.7 Hz, 1H), 8.27 (s, 1H), 7.63-7.56 (m, 1H), 7.42-7.36 (m, 1H), 7.16 (dd, J=8.6, 3.9 Hz, 1H), 6.93 (dd, J=10.2 , 8.6 Hz, 1H), 4.71 (d, J=1.6 Hz, 2H), 3.16-3.11 (m, 6H), 3.07-3.03 (m, 2H) .

HPLC-MS (Method D): [m/z]: 461.1 [M+H]+.

2-(2-{[2-(4,6-Difluoro-1H-1,3-benzodiazol-2-yl)ethyl]amino}ethyl)-N-[(3-fluoropyridin-2-yl)methyl]1 ,3-oxazole-4-carboxamide (illustrative example No. 249)

- 176 039916

Similar to general procedure 8, 2-(2-Aminoethyl)-N-[(3-fluoropyridin-2-yl)methyl]-1,3-oxazole-4-carboxamide dihydrochloride (235) (500 mg, 1.48 mmol), N -(2,4-difluoro-6-nitrophenyl)prop-2-enamide (K4) (338 mg, 1.48 mmol) and DBU (0.664 ml, 4.45 mmol) in MeCN (15 ml) gave the crude intermediate , which was partially purified by flash column chromatography using a gradient of 0-20% MeOH in DCM. The resulting intermediate was further reacted with iron powder (145 mg) in AcOH (4 mL) to give the title compound (23 mg, 8%) as an off-white solid after purification by flash column chromatography (eluting with a 0- 40% MeOH/DCM), followed by kp-NH flash chromatography (eluting with a gradient of 0-5% MeOH/DCM).

¹ H NMR (DMSO-ib, 500 MHz): δ [ppm] = 8.53-8.46 (m, 2H), 8.41-8.36 (m, 1H), 7.73- 7.67 (m, 1H), 7.437.37 (m, 1H), 7.14 (dd, J=8.9, 2.2 Hz, 1H), 6.96 (td, J=10.7, 2.2 Hz, 1H), 4.62 (dd, J=5.7, 1.4 Hz, 2H), 3.02-2.90 (m, 8H).

HPLC-MS (method d): [m/z]: 445.2 [M+H]+.

2-(2-{[2-(5-Fluoro-1H-1,3-benzodiazol-2-yl)ethyl]amino}ethyl)-N-[(3-fluoropyridin-2-yl)methyl]-1, 3oxazole-4-carboxamide (illustrative example No. 250)

Similar to general procedure 8, 2-(2-aminoethyl)-I-[(3-fluoropyridin-2-yl)methyl]-1,3-oxazole-4-carboxamide dihydrochloride (235) (1.07 g, 2.59 mmol, 82% pure), N-(5-fluoro-2-nitrophenyl)prop-2enamide (J) (0.54 g, 2.59 mmol) and DBU (1.28 ml, 8.54 mmol) in MeCN (30 ml) at room temperature for 20 hours gave a mixture of mono:bis-alkylated (1:1.4) adducts (1.1 g) which was reacted with iron powder (0.52 g) in AcOH (8 ml) to give the title compound (90 mg, 9%) as a brown solid after purification by flash column chromatography (elution gradient 0-30% MeOH/DCM), followed by kp-NH flash column chromatography (elution gradient 0 -2% MeOH/DCM), followed by flash column chromatography (eluting with a 0-20% MeOH/DCM gradient) and basic prep-HPLC.

1H NMR (DMSO-H6, 500 MHz): δ [ppm] = 8.52-8.47 (m, 2H), 8.39-8.36 (m, 1H), 7.72-7 .66 (m, 1H), 7.467.36 (m, 2H), 7.24 (dd, J=9.6, 2.3 Hz, 1H), 6.99-6.89 (m, 1H), 4.61 (dd, J=5.7, 1.3 Hz, 2H), 3.02-2.89 (m, 8H).

HPLC-MS (Method C): [m/z]: 427.2 [M+H] ⁺ .

2-(2-{[2-(4,7-Difluoro-1H-1,3-benzodiazol-2-yl)ethyl]amino}ethyl)-X-[(3-fluoropyridin-2-yl)methyl]1 ,3-oxazole-4-carboxamide (illustrative example no. 251)

Similar to general procedure 8, 2-(2-aminoethyl)-N-[(3-fluoropyridin-2-yl)methyl]-1,3-oxazole-4-carboxamide dihydrochloride (235) (400 mg, 1.19 mmol), N -(3,6-difluoro-2-nitrophenyl)prop-2-enamide (K6) (316 mg, 1.20 mmol) and DBU (0.53 ml, 3.56 mmol) in MeCN (15 ml) gave crude an intermediate which was partially purified by flash column chromatography eluting with a gradient of 0-20% MeOH in DCM. This intermediate was further reacted with iron powder (230 mg) in AcOH (4 ml) to give the title compound (63 mg, 14%) as an off-white solid after purification by flash column chromatography (eluting with a 0- 40% MeOH/DCM), followed by kp-NH flash chromatography (eluting with a gradient of 0-5% MeOH/DCM).

1H NMR (DMSO-H6, 500 MHz): δ [ppm] = 8.52-8.46 (m, 2H), 8.38 (m, 1H), 7.74-7.66 (m , 1H), 7.44-7.37 (m, 1H), 6.95-6.91 (m, 2H), 4.62 (dd, J=5.6, 1.4 Hz, 2H), 3.03-2.90 (m, 8H).

HPLC-MS (method d): [m/z]: 445.2 [M+H] ⁺ .

- 177 039916 N[(3-fluoropyridin-2-yl)methyl]-2-[2-({2-[4-(trifluoromethyl)-1H-1,3-benzodiazol-2yl]ethyl}amino)ethyl]- 1,3-oxazole-4-carboxamide (illustrative example No. 256)

Similar to general procedure 8, 2-(2-aminoethyl)-N-[(3-fluoropyridin-2-yl)methyl]-1,3-oxazole-4-carboxamide dihydrochloride (235) (300 mg, 0.89 mmol), no. [2-nitro-6-(trifluoromethyl)phenyl]prop-2enamide (K8) (272 mg, 0.90 mmol) and DBU (398 µl, 2.67 mmol) in MeCN (10 ml) gave a crude intermediate which partially purified by flash column chromatography using a gradient of 0-10% MeOH in DCM. The resulting intermediate was further reacted with iron powder (177 mg) in AcOH (3 mL) to give the title compound (99 mg, 26%) as a yellow solid after purification by flash column chromatography (eluting with a 0-25% gradient). MeOH/DCM), followed by kp-NH flash chromatography (eluting with a gradient of 0-6% MeOH/DCM).

1H NMR (DMSO-06, 500 MHz): δ [ppm] = 8.53-8.47 (m, 2H), 8.40-8.37 (m, 1H), 7.75 (d , J=8.0 Hz, 1H), 7.73-7.67 (m, 1h), 7.45 (d, J=7.6 Hz, 1H), 7.43-7.38 (m, 1H), 7.28 (t, J=7.8 Hz, 1H), 4.62 (d, J=4.7 Hz, 2H), 3.01 (s, 4H), 3.00-2, 93 (m, 4H).

HPLC-MS (method d): [m/z]: 477.4 [M+H]+.

2-(2-{[2-(4-fluoro-1H-1,3-benzodiazol-2-yl)ethyl]amino}ethyl)-N-(pyridin-2-ylmethyl)-1,3-oxazol-4carboxamide (illustrative example #236)

Similar to general procedure 8, 2-(2-aminoethyl)-N-(pyridin-2-ylmethyl)-1,3-oxazol-4-carboxamide dihydrochloride (236) (350 mg, 0.95 mmol), N-(3 -fluoro-2-nitrophenyl)prop-2-enamide (G) (206 mg, 0.95 mmol), DBU (1.0 ml, 2.85 mmol) in MeCN (15 ml) gave a mixture which was partially purified by the method flash column chromatography (eluting with a 0-15% MeOH/DCM gradient). The resulting mixture was further reacted with iron powder (136 mg) in AcOH (4 mL) to give the title compound (9 mg, 4%) as a pale brown solid after purification by flash column chromatography (gradient elution 0-40% MeOH/DCM), kp-NH flash column chromatography (elution with 0-10% MeOH/DCM gradient) and basic prep-HPLC.

1H NMR (MeOD, 500 MHz): δ [ppm] = 8.52-8.47 (m, 1H), 8.28 (s, 1H), 7.80 (td, J=7.7 , 1.8 Hz, 1H), 7.39 (d, J=7.9 Hz, 1H), 7.34-7.30 (m, 1H), 7.28 (d, J=8.0 Hz , 1H), 7.19-7.10 (m, 1H), 6.96-6.88 (m, 1H), 4.65 (s, 2H), 3.17-3.08 (m, 6H ), 3.08-3.02 (m, 2H).

HPLC-MS (Method D): [m/z]: 409.1 [M+H] ⁺ .

Methyl 2-(3-{[(tert-butoxy)carbonyl]amino}propanamido)-3-hydroxybutanoate (315)

Similar to general procedure 13, methyl L-threonate hydrochloride (1:1) (11 g, 64.9 mmol), N-(tert-butoxycarbonyl)-beta-alanine (12.3 g, 64.9 mmol), TEA (9, 94 ml, 71.3 mmol) and DCC (14.7 g, 71.3 mmol) in DCM (250 ml) gave the title compound (16.4 g, 72.4%, 87% pure) as a viscous yellow oil after purification by flash column chromatography (eluting with a 0-100% EtOAc/heptane gradient).

1H NMR (CDCl ₃ , 500 MHz): δ [ppm] = 6.35 (s, 1H), 5.12 (s, 1H), 4.59 (dd, J=8.8, 2, 5 Hz, 1H), 4.36 (q.d., J=6.4, 2.5 Hz, 1H), 3.78 (s, 3H), 3.45 (q.d., J=5.9 Hz, 2H), 2.57-2.46 (m, 2H), 1.43 (s, 9H), 1.23 (d, J=6.4 Hz, 3H).

HPLC-MS (method A): [m/z]: 291.05 [M+H] ⁺ .

Methyl 2-(2-{[(tert-butoxy)carbonyl]amino}ethyl)-5-methyl-4,5-dihydro-1,3-oxazole-4-carboxylate (316)

Similar to general procedure 14, methyl 2-(3-{[(tert-butoxy)carbonyl]amino}propanamido)-3hydroxybutanoate (315) (1 g, 2.87 mmol, 87% pure), DAST (0.45 ml, 3.44 mmol) and K ₂ CO ₃ (0.79 g, 5.73 mmol) in anhydrous DCM (25 mL) gave the title compound (0.61 g, 67%) as a pale yellow oil after column purification. flash chromatography (gradient elution 0-178 039916

100% EtOAc/heptane).

¹ H NMR (CDCl ₃ , 250 MHz): δ [ppm] = 5.22 (brs, 1H), 4.87 (d.s., J=9.9, 6.3 Hz, 1H), 4.76 (dt,

J=10.1, 1.2 Hz, 1H), 3.76 (s, 3H), 3.52-3.37 (m, 2H), 2.53-2.44 (m, 2H), 1 .44 (s, 9H), 1.29 (d, J=6.3 Hz, 3H).

Methyl 2-(2-{[(tert-butoxy)carbonyl]amino}ethyl)-5-methyl-1,3-oxazole-4-carboxylate (317) o

Ao'—NHBoc

As in general procedure 15, methyl 2-(2-{[(tert-butoxy)carbonyl]amino}ethyl)-5-methyl-4,5dihydro-1,3-oxazole-4-carboxylate (316) (0.61 g , 1.98 mmol), bromo(trichloro)methane (0.59 ml, 5.95 mmol) and DBU (0.89 ml, 5.95 mmol) in DCM (15 ml) gave the title compound (0. 42 g, 71%) as a dark orange oil which solidified on standing after purification by flash column chromatography (eluting with a 0-100% EtOAc/heptane gradient).

¹ H NMR (CDCl ₃ , 250 MHz): δ [ppm] = 4.94 (s, 1H), 3.90 (s, 3H), 3.55 (q, J=6.2 Hz, 2H), 2.93 (t, J=6.3 Hz, 2H), 2.59 (s, 3H), 1.42 (s, 9H).

HPLC-MS (Method A): [m/z]: 284.95 [M+H]+.

2-(2-{[(tert-Butoxy)carbonyl]amino}ethyl)-5-methyl-1,3-oxazole-4-carboxylic acid (318) o

ΗοΑγΝ_ /А-О '—NHBoc

Similar to general procedure 5, methyl 2-(2-{[(tert-butoxy)carbonyl]amino}ethyl)-5-methyl-1,3oxazole-4-carboxylate (317) (0.42 g, 1.42 mmol) , LiOH (0.067 g, 2.84 mmol) in THF (8 ml) and water (2 ml) gave the title compound (0.4 g, 98%) as a pale orange oil which solidified on standing.

1H NMR (DMSO-d ₆ , 250 MHz): δ [ppm] = 12.72 (s, 1H), 6.90 (t, J=5.5 Hz, 1H), 3.25 (d , J=6.2 Hz, 2H), 2.79 (t, J=6.9 Hz, 2H), 1.34 (s, 9H) [CH3 peak, overlap with DMSO signals].

HPLC-MS (method A): [m/z]: 293.00 [M+Na]+.

tert-Butyl N-[2-(4-{[(3-fluoropyridin-2-yl)methyl]carbamoyl}-5-methyl-1,3-oxazol-2yl)ethyl]carbamate (319) ^NHBoc

Similar to general procedure 6, 2-(2-{[(tert-butoxy)carbonyl]amino}ethyl)-5-methyl-1,3-oxazole-4carboxylic acid (318) (0.4 g, 1.39 mmol) , (3-fluoropyridin-2-yl)methanamine dihydrochloride (A2) (0.415 g, 2.09 mmol), DIPEA (0.8 ml, 4.59 mmol) and HATU (0.79 g, 2.09 mmol) in DMF (6 mL) gave the title compound (0.653 g, 79, 64% pure) as a yellow oil after purification by flash column chromatography (eluting with a 0-100% EtOAc/heptane gradient).

1H NMR (DMSO-b ₆ , 250 MHz): δ [ppm] = 8.44-8.30 (m, 2H), 7.75-7.64 (m, 1H), 7.46- 7.34 (m, 1H), 6.97 (brs, 1H), 4.60 (d, J=4.2 Hz, 2H), 3.34-3.24 (m, 2H), 2 .83 (t, J=6.8 Hz, 2H), 1.35 (s, 9H).

HPLC-MS (method A): [m/z]: 379.05 [M+H] ⁺ .

2-(2-Aminoethyl)-N-[(3-fluoropyridin-2-yl)methyl]-5-methyl-1,3-oxazole-4-carboxamide dihydrochloride (320)

1 ⁰ /^0 ^νη ₂

2HCI

Similar to general procedure 2, tert-butyl N-[2-(4-{[(3-fluoropyridin-2-yl)methyl]carbamoyl}-5methyl-1,3-oxazol-2-yl)ethyl]carbamate (319) (0.65 g, 1.10 mmol, 64% purity) and 12M HC (1.16 ml, 11.04 mmol) in MeOH (10 ml) gave the title compound (0.51 g, 76% purity) as a hygroscopic off-white solid. The compound was used in the next step without further purification.

¹ H NMR (MeOD, 250 MHz): δ [ppm] = 8.55 (dd, J=5.3, 1.0 Hz, 1H), 8.22-8.10 (m, 1H) , 7.86-7.75 (m, 1H), 3.70 (dd, J=11.2, 7.3 Hz, 3H), 3.43 (t, J=6.5 Hz, 2H), 3.17 (t, J=6.5 Hz, 2H), 2.58 (s, 3H) [CH ₂ peak, overlap with H ₂ O signals].

HPLC-MS (method A): [m/z]: 279.2 [M+H] ⁺ .

N-[(3-fluoropyridin-2-yl)methyl]-5-methyl-2-[2-({2-[(2-nitrophenyl)carbamoyl]ethyl}amino)ethyl]-1,3oxazole-4-carboxamide (illustrative example #230)

- 179 039916

Similar to general procedure 8, 2-(2-aminoethyl)-N-[(3-fluoropyridin-2-yl)methyl]-5-methyl-1,3oxazole-4-carboxamide dihydrochloride (320) (0.51 g, 1 .11 mmol, 76% pure), N-(2-nitrophenyl)prop-2enamide (D) (0.4 g, 2.09 mmol) and DBU (0.69 ml, 4.59 mmol) in MeCN (15 ml) at room temperature for 20 hours gave a mixture of mono:bis-alkylated (1:3.6) adducts which was reacted with iron powder (0.31 g) in AcOH (3.5 ml) to give the title compound (0.12 g, 20%) as a light green solid after purification by flash column chromatography (elution gradient 0-30% MeOH/DCM), followed by flash column chromatography (KP-NH, elution gradient 0- 5% MeOH/DCM), and final purification by flash column chromatography (eluting with a smooth gradient of 0-30% MeOH/DCM).

1H NMR (MeOD, 250 MHz): δ [ppm] = 8.33-8.30 (d, J=4.7 Hz, 1H), 7.60-7.54 (m, 1H), 7.48-7.40 (m, 2H), 7.38-7.33 (m, 1H), 7.18-7.14 (m, 2H), 4.65 (d, J=1.6 Hz, 2H), 3.15-3.07 (m, 6H), 2.97 (t, J=6.6 Hz, 2H), 2.52 (s, 3H).

HPLC-MS (Method C): [m/z]: 423.1 [M+H]+.

The above general scheme 12:

2-(2-{[2-(5-cyano-1H-1,3-benzodiazol-2-yl)ethyl]αmino}ethyl)-N-[(3-fluoropyridin-2-yl)methyl]-1, 3oxazole-4-carboxamide (illustrative example No. 265) ⁴

AjAcN

Similar to general procedure 8, 2-(2-aminoethyl)-N-[(3-fluoropyridin-2-yl)methyl]-1,3-oxazole-4-carboxamide dihydrochloride (235) (400 mg, 1.19 mmol) and N -(4-cyano-2-nitrophenyl)prop-2-enamide (K12) (325 mg, 1.49 mmol) and DBU (0.53 ml, 3.56 mmol) in MeCN (12 ml) gave the crude intermediate , which reacted with iron powder (239 mg, 4.28 mmol) in AcOH (4 mL) to give the title compound (24 mg, 5%) as a brown solid after purification by flash column chromatography (eluting with a 0- 20% MeOH/DCM).

1H NMR (DMSO-d6, 500 MHz): δ [ppm] = 8.51-8.46 (m, 2H), 8.39-8.36 (m, 1H), 8.01-7 .95 (m, 1H), 7.727.67 (m, 1H), 7.63-7.59 (m, 1H), 7.50 (dd, J=8.3, 1.5 Hz, 1H), 7.42-7.38 (m, 1H), 4.63-4.59 (m, 2H), 3.02-2.91 (m, 8H).

HPLC-MS (method B): [m/z]: 434.2 [M+H] ⁺ .

The above general scheme 13:

methyl 1-(cyanomethyl)-1H-1,2,4-triazole-3-carboxylate (239)

Methyl 1H-1,2,4-triazole-3-carboxylate (5 g, 39.34 mmol) and 2-bromoacetonitrile (2.7 ml, 39.34 mmol) in DMF (45 ml) were stirred for 5 minutes. K ₂ CO ₃ (5.44 g, 39.34 mmol) was added and the reaction mixture was stirred at room temperature for 23 h. The solvent was evaporated in vacuo, water (20 ml) was added and the aqueous layer was extracted with ethyl acetate (8x50 ml) . The combined organic layers were washed with 5% aqueous LiCl solution (2x20 ml), saturated sodium chloride solution (2x20 ml), dried (MgSO ₄ ), filtered and evaporated to give a mixture of regioisomers (1.5:1) as a dark brown oils (5.27 g). The resulting crude product was purified by flash column chromatography (eluting with a gradient of 10 to 100% EtOAc/heptane) to give the title compound (3.5 g, 45%) as a pale brown oil which solidified on standing.

1 H NMR (DMSOCH, 250 MHz): δ [ppm] = 8.30 (s, 1H), 5.79 (s, 2H), 3.94 (s, 3H).

Methyl 1-(2-{[(tert-butoxy)carbonyl]amino}ethyl)-1H-1,2,4-triazole-3-carboxylate (240) o

MeO^V^N.

I N

NHBoc

Methyl 1-(cyanomethyl)-1H-1,2,4-triazole-3-carboxylate (239) (1 g, 6.02 mmol), di-tert-butyl dicarbonate (2.63 g, 12.04 mmol) and TEA (1.7 ml, 12.04 mmol) in EtOH (30 ml) were stirred for 5 min, then 10% palladium on charcoal (128 mg, 0.602 mmol) was added. The reaction mixture was stirred under an atmosphere

- 180 039916 hydrogen 18 hours the Reaction mixture was filtered through Celite, and the resulting residue was washed with EtOH (30 ml). The filtrate was evaporated to give a brown oil which crystallized on standing (1.95 g).

Purification by flash column chromatography (eluting with a 0-100% EtOAc/heptane gradient) gave the title compound (1.35 g, 69.7%) as a pale brown solid.

¹ H NMR (CDCl ₃ , 250 MHz): δ [ppm] = 8.13 (s, 1H), 4.69 (br s, 1H), 4.40 (t, J=5.6 Hz, 2H), 4.00 (s, 3H), 3.60 (pseudo q, J=5.9 Hz, 2H), 1.42 (s, 9H).

HPLC-MS (method A): [m/z]: 271.1 [M+H]+.

-(2-{[(tert-Butoxy)carbonyl]amino}ethyl)-1H-1,2,4-triazole-3-carboxylic acid (241) o ^NHBoc

Similarly, using General Procedure 5, LiOH (0.11 g, 4.72 mmol) and methyl 1-(2{[(tert-butoxy)carbonyl]amino}ethyl)-1H-1,2,4-triazole- 3-carboxylate (240) (0.87 g, 2.69 mmol) in THF (20 mL)/water (5 mL) gave the title compound (0.71 g, 100%) as a cream colored solid . The compound was used in the next step without further purification.

1H NMR (DMSO-d6, 250 MHz): δ [ppm] = 8.54 (s, 1H), 6.97 (br s, 1H), 4.26 (t, J=5.9 Hz, 2H), 3.34 (pseudo q, J=5.8 Hz, 2H) overlap with water peak, 1.34 (s, 9H).

HPLC-MS (method A): [m/z]: 257.05 [M+H] ⁺ .

tert-Butyl N-[2-(3-{[(3-fluoropyridin-2-yl)methyl]carbamoyl}-1H-1,2,4-triazol-1-yl)ethyl]carbamate (242)

F about

UJ, n । No.=/A _NHBoc

Similarly, using General Procedure 6, 1-(2-{[(tert-butoxy)carbonyl]amino}ethyl)1H-1,2,4-triazole-3-carboxylic acid (241) (0.5 g, 1 .95 mmol), (3-fluoropyridin-2-yl)methanamine dihydrochloride (A2) (0.58 g, 2.93 mmol), DIPEA (1.7 ml, 9.76 mmol), HATU (1.11 g , 2.93 mmol) in DMF (10 mL) gave the title compound (0.55 g, 76%) as an off-white solid after purification by flash column chromatography (eluting with a gradient of 0-10% MeOH/ DCM), followed by a second purification by flash column chromatography (eluting with a gradient of 010% MeOH/EtOAc).

1H NMR (DMSO-d ₆ , 500 MHz): δ [ppm] = 8.74 (t, J=5.6 Hz, 1H), 8.57 (s, 1H), 8.39 (d , J=4.6 Hz, 1H), 7.78-7.63 (m, 1H), 7.42 (dt, J=8.6, 4.4 Hz, 1H), 7.00 (s, 1H), 4.64 (d, J=4.7 Hz, 2H), 4.28 (t, J=5.9 Hz, 2H), 3.36 (pseudo q, J=5.9 Hz, 2H ), 1.35 (s, 9H).

HPLC-MS (method A): [m/z]: 365.15 [M+H] ⁺ .

tert-Butyl N-[2-(3-{[(3-chloropyridin-2-yl)methyl]carbamoyl}-1H-1,2,4-triazol-1-yl)ethyl]carbamate (243)

CI O

I । n I

A _NHBoc

Similarly, using General Procedure 6, 1-(2-{[(tert-butoxy)carbonyl]amino}ethyl)1H-1,2,4-triazole-3-carboxylic acid (241) (0.58 g, 2 .26 mmol), (3-chloropyridin-2-yl)methanamine dihydrochloride (0.732 g, 3.4 mmol), HATU (1.29 g, 3.4 mmol), DIPEA (1971 µl, 11.32 mmol) in DMF (11 mL) gave the title compound (0.87 g, 96%) as a brown foam after purification by flash column chromatography eluting with a 0-10% MeOH/DCM gradient.

1H NMR (DMSO-d6, 500 MHz): δ [ppm] = 8.73 (t, J=5.3 Hz, 1H), 8.57 (s, 1H), 8.52 (d, J=4.4 Hz, 1H), 7.96 (d, J=8.0 Hz, 1H), 7.39 (dd, J=8.0, 4.7 Hz, 1H), 7.00 ( s, 1H), 4.67 (d, J=5.4 Hz, 2H), 4.28 (t, J=5.9 Hz, 2H), 3.36 (pseudo q, J=6.1 Hz , 2H), 1.35 (s, 9H).

HPLC-MS (method A): [m/z]: 381.05 [M+H] ⁺ .

-(2-Aminoethyl)-N -[(3-fluoropyridin-2-yl)methyl]-1H-1,2,4-triazole-3-carboxamide dihydrochloride (244)

Similarly, using the general procedure 2, 12 M HC (2.48 ml) and tert-butyl N-[2-(3-{[(3fluoropyridin-2-yl)methyl]carbamoyl}-1H-1,2,4 -triazol-1-yl)ethyl]carbamate (242) (0.54 g, 1.49 mmol) in MeOH (10 ml) at 40°C for 2 h gave the title compound (0.52 g, 100 %) as an off-white foam. The compound was used in the next step without further purification.

¹ H NMR (DMSO-d ₆ , 250 MHz): δ [ppm] = 8.89 (t, J=5.6 Hz, 1H), 8.70 (s, 1H), 8.39 ( d, J=4.7 Hz, 1H), 8.25 (s, 3H), 7.80-7.67 (m, 1H), 7.43 (dt, J=8.6, 4.5 Hz , 1H), 4.66 (d, J=4.4 Hz, 2H), 4.55 (t, J=5.9 Hz, 2H), 3.32

- 181 039916 (kv, J=5.6 Hz, 2H).

HPLC-MS (method A): [m/z]: 265.05 [M+H]+.

-(2-Aminoethyl)-N-[(3-chloropyridin-2-yl)methyl]-1H-1,2,4-triazole-3-carboxamide dihydrochloride (245)

Similarly, using General Procedure 2, 12M HC (3.6 mL) and t-butyl N-[2-(3-{[(3chloropyridin-2-yl)methyl]carbamoyl}-1H-1,2,4- triazol-1-yl)ethyl]carbamate (243) (0.87 g, 2.17 mmol) in MeOH (15 ml) at 40°C for 3 h gave the title compound (0.84g, 99%) in the form of light orange foam.

1H NMR (DMSO-d ₆ , 250 MHz): δ [ppm] = 8.89 (t, J=5.5 Hz, 1H), 8.74 (s, 1H), 8.53 (dd , J=4.7, 1.3 Hz, 1H), 8.35 (s, 3H), 7.99 (dd, J=8.1, 1.4 Hz, 1H), 7.42 (dd, J=8.1, 4.7 Hz, 1H), 4.69 (d, J=5.5 Hz, 2H), 4.58 (t, J=5.9 Hz, 2H), 3.33 ( q, J=5.8 Hz, 2H), 3.17 (s, 1H), 2.69 (s, 1H).

HPLC-MS (Method A): [m/z]: 280.95 [M+H] ⁺ .

1-{2-[(1H-1,3-Benzodiazol-2-ylmethyl)amino]ethyl} -N-[(3-fluoropyridin-2-yl)methyl]-1H-1,2,4triazol-3- carboxamide (illustrative example No. 169) °

Ui ^N

Η

Similarly, using General Procedure 3, 1H-1,3-benzodiazol-2-carbaldehyde (104 mg, 0.71 mmol), 1-(2-aminoethyl)-N-[(3-fluoropyridin-2-yl)methyl ]-1H-1,2,4-triazole-3-carboxamide dihydrochloride (244) (200 mg, 0.59 mmol) and DIPEA (0.41 ml, 2.37 mmol) in MeOH (8 ml) at room temperature over 17 hours, followed by the addition of NaBH ₄ (32 mg, 0.86 mmol) gave the title compound (79 mg, 34%) as a beige solid after purification by basic pre-HPLC.

1H NMR (DMSO-d ₆ , 500 MHz): δ [ppm] = 12.16 (s, 1H), 8.76 (t, J=5.6 Hz, 1H), 8.65 (s , 1H), 8.38 (d, J=4.7 Hz, 1H), 7.70 (pseudo t, J=9.3 Hz, 1H), 7.49 (broad d, J=38.6 Hz, 2H), 7.40-7.43 (m, 1H), 7.12 (d, J=3.7 Hz, 2H), 4.64 (d, J=5.5 Hz, 2H), 4.34 (t, J=6.0 Hz, 2H), 3.92 (s, 2H), 3.02 (t, J=5.9 Hz, 2H).

HPLC-MS (Method C): [m/z]: 395.2 [M+H] ⁺ .

1-(2-{[2-(1H-1,3-Benzodiazol-2-yl)ethyl]amino}ethyl)-N-[(3-fluoropyridin-2-yl)methyl]-1H-1,2, 4triazole-3-carboxamide (illustrative example No. 181)

Similarly, using General Procedure 8, DBU (0.39 ml, 2.58 mmol), 1-(2-aminoethyl)N-[(3-fluoropyridin-2-yl)methyl]-1H-1,2,4 -triazole-3-carboxamide dihydrochloride (244) (300 mg, 0.86 mmol) and N-(2-nitrophenyl) prop-2-enamide (D) (165 mg, 0.86 mmol) in MeCN (14 ml) at room temperature for 16 hours gave the crude intermediate (0.364 g) as a mixture of mono:bis alkylated (2:1) adducts. The resulting mixture was dissolved in AcOH (5 ml), iron (112 mg) was added and the reaction mixture was heated at 80° C. for 4 h to give the title compound (20 mg, 11%) as an off-white solid after purification by basic prep-HPLC, followed by flash column chromatography (x2) (KP-NH, eluting with a gradient of 0-30% MeOH/DCM and 5-20% MeOH/DCM).

Ή NMR (DMSO-d ₆ , 500 MHz) δ [ppm] = 12.14 (s, 1H), 8.75 (t, J=5.6 Hz, 1H), 8.59 (s, 1H), 8.41-8.35(m, 1H), 7.73-7.67(m, 1H), 7.45(brs, 2H), 7.43-7.38(m, 1H), 7.13-7.07 (m, 2H), 4.64 (dd, J=5.6, 1.3 Hz, 2H), 4.31 (t, J=6.0 Hz, 2H ), 2.99 (dt, J=12.3, 6.0 Hz, 4H), 2.94-2.89 (m, 2H).

HPLC-MS (method C): [m/z]: 409.3 [M+H]+.

1-{2-[(1H-1,3-Benzodiazol-2-ylmethyl)amino]ethyl}-N-[(3-chloropyridin-2-yl)methyl]-1H-1,2,4triazol-3-carboxamide (illustrative example #170)

- 182 039916

Similarly, using General Procedure 3, 1H-1,3-Benzodiazol-2-carbaldehyde (156 mg, 1.07 mmol), 1-(2-aminoethyl)-N-[(3-chloropyridin-2-yl)methyl ]-1H-1,2,4-triazole-3-carboxamide dihydrochloride (245) (350 mg, 0.89 mmol) and DIPEA (0.62 ml, 3.56 mmol) in MeOH (10 ml) at 40° C for 2 h followed by addition of NaBH ₄ (50.5 mg, 1.34 mmol) gave the title compound (190 mg, 52%) as a cream solid after purification by basic prep-HPLC.

1H NMR (DMSO-a ₆ , 500 MHz): δ [ppm] = 12.16 (s, 1H), 8.75 (t, J=5.4 Hz, 1H), 8.66 (s , 1H), 8.51 (dd, J=4.7, 1.1 Hz, 1H), 7.95 (dd, J=8.1, 1.2 Hz, 1H), 7.49 (br. d, J=37.6 Hz, 2H), 7.39 (dd, J=8.0, 4.7 Hz, 1H), 7.12 (s, 2H), 4.67 (d, J=5 .4 Hz, 2H), 4.35 (t, J=6.0 Hz, 2H), 3.93 (s, 2H), 3.03 (t, J=5.9 Hz, 2H).

HPLC-MS (method C): [m/z]: 411.3 [M+H] ⁺ .

The above general scheme 14:

methyl 1-(2-{[(tert-butoxy)carbonyl]amino}ethyl)-1H-pyrazole-4-carboxylate (246)

DIAD (9.39 ml, 43.61 mmol) was added dropwise to an ice-cold solution of methyl 1H-pyrazole-4-carboxylate (5 g, 39.65 mmol), tert-butyl (2-hydroxyethyl) carbamate (6.13 ml, 39 .65 mmol) and PPh ₃ (11.44 g, 43.61 mmol) in THF (300 ml). The reaction mixture was then allowed to warm to room temperature and stirred for 1.5 hours. The reaction was quenched with water (150 ml) and the layers were separated. The aqueous layer was extracted with ethyl acetate (3x150 ml). The combined organic layers were washed with brine, dried (MgSO4), filtered and the solvent was evaporated to give the crude product. Purification by flash column chromatography (eluting with a 15-100% EtOAc/heptane gradient) gave the title compound (11.3 g, 68%) as a white solid.

1H NMR (CDCl ₃ , 500 MHz): δ [ppm] = 7.93 (s, 1H), 7.87 (s, 1H), 4.25 (t, J=5.3 Hz, 2H ), 3.83 (s, 3H), 3.61-3.54 (m, 2H), 1.44 (s, 9H).

HPLC-MS (method A): [m/z]: 270.1 [M+H] ⁺ .

Ethyl 1-(cyanomethyl)-3,5-dimethyl-1H-pyrazole-4-carboxylate (247)

A mixture of ethyl 3,5-dimethyl-1H-nirazole-4-carboxylate (2 g, 11.89 mmol), bromoacetonitrile (1.24 ml, 17.84 mmol), K ₂ CO ₃ (3.29 g, 23, 78 mmol) and acetone (6 ml) were heated at 100°C in a microwave oven for 1 hour. More bromoacetonitrile (0.828 ml, 11.89 mmol) was added and the resulting mixture was heated at 100°C in a microwave oven for another 1 hour. The reaction mixture was evaporated in vacuo, the resulting residue was dissolved in water (100 ml) and extracted with ethyl acetate (5x50 ml) . The combined organic extracts were dried (MgSO ₄ ), filtered and evaporated in vacuo. Purification by flash column chromatography eluting with a 0-20% MeOH/DCM gradient gave the title compound (2.22 g, 71%) as an orange solid.

1H NMR (DMSOCH, 500 MHz): δ [ppm] = 5.42 (s, 2H), 4.21 (q, J=7.1 Hz, 2H), 2.50 (s, 3H) , 2.30 (s, 3H), 1.28 (t, J=7.1 Hz, 3H).

HPLC-MS (Method A): [m/z]: 207.95 [M+H]+.

Ethyl 1-(2-{[(tert-butoxy)carbonyl]amino}ethyl)-3,5-dimethyl-1H-pyrazole-4-carboxylate (248)

A suspension of ethyl 1-(cyanomethyl)-3,5-dimethyl-1H-pirazole-4-carboxylate (247) (2.22 g, 9.64 mmol), BOC anhydride (4.21 g, 19.27 mmol), TEA (2.69 ml, 19.27 mmol) and palladium-carbon (10% wt, 0.103 g, 0.964 mmol) in EtOH (100 ml) were stirred under hydrogen atmosphere for 28 hours. The reaction mixture was filtered and the resulting residue was washed with MeOH. The combined filtrates were evaporated in vacuo and the resulting residue was partitioned between water (100 ml) and EtOAc (50 ml). The phases were separated, the aqueous phase was extracted with ethyl acetate (3x50 ml). The combined organic layers were dried (MgSO ₄ ), filtered and evaporated in vacuo. Purification by flash column chromatography eluting with a 70-100% EtOAc/heptane gradient gave the title compound (1.52 g, 51%) as a tan solid.

1H NMR (DMSO-06, 500 MHz): δ [ppm] = 6.90 (s, 1H), 4.18 (q, J=7.1 Hz, 2H), 4.02 (t, J=6.1Hz, 2H), 3.21(q, J=6.1Hz, 2H), 2.42(s, 3H), 2.27(s, 3H), 1.38-1, 28 (s, 9H), 1.26 (t, J=7.1 Hz, 3H).

HPLC-MS (method A): [m/z]: 312.45 [M+H] ⁺ .

- 183 039916

Ethyl (248)

1-(2-{[(tert-butoxy)carbonyl]amino}ethyl)-3-(trifluoromethyl)-III-pyrazole-4-carboxylate

Solution

ethyl 3-(trifluoromethyl)-1H-pyrazole-4-carboxylate (3 g, 14.41 mmol), tert-butyl (2 hydroxyethyl) carbamate (2.45 ml, 15.85 mmol) and PPh ₃ (4.16 g, 15.85 mmol) in THF (50 ml) was stirred at room temperature for 16 h. More (2-hydroxyethyl)carbamate (2.45 ml, 15.85 mmol) was added and the reaction mixture was stirred at room temperature in within 48 hours the Reaction was extinguished with water (30 ml) and was extracted with EtOAc (3x50 ml). The combined organic layers were washed with saturated sodium chloride solution (30 ml), dried (MgSO ₄ ), filtered and evaporated to give a yellow oil. Purification by flash column chromatography (elution with a 5-80% TBME/heptane gradient) gave a residue which was repurified by flash column chromatography (elution with a 5-60% TBME/heptane gradient) to give the title compound (1, 19 g, 21%) as a white solid.

1H NMR (CDCl3, 500 MHz): δ [ppm] = 7.97 (s, 1H), 4.72 (broad s, 1H), 4.32 (q, J=7.1 Hz, 2H), 4.32-4.27 (m, 2H), 3.59 (pseudo q, J=5.9 Hz, 2H), 1.44 (s, 9H), 1.35 (t, J= 7.1 Hz, 3H).

HPLC-MS (method A): [m/z]: 374 [M+Na]+.

1-(2-{[(tert-Butoxy)carbonyl]amino}ethyl)-1H-pyrazole-4-carboxylic acid (249)

Similar to general procedure 5, LiOH (0.72 g, 30.08 mmol) and 1-(2-{[(tert-butoxy)carbonyl]amino}ethyl)-1H-pyrazole-4-carboxylate (246) (54%, 5 g, 10.03 mmol) in THF (80 ml) and water (30 ml) at room temperature for 24 hours gave the title compound (2.6 g, 96.5%). The compound was used further without purification.

1H NMR (Methanol-b ₄ , 500 MHz): δ [ppm] = 8.06 (s, 1H), 7.87 (s, 1H), 4.23 (t, J=5.8 Hz , 2H), 3.46 (pseudo q, J=5.5 Hz, 2H), 1.42 (d, J=19.0 Hz, 9H).

HPLC-MS (method A): [m/z]: 255.95 [M+H] ⁺ .

1-(2-{[(tert-Butoxy)carbonyl]amino}ethyl)-3,5-dimethyl-1H-pyrazole-4-carboxylic acid (250)

Similar to general procedure 5, ethyl 1-(2-{[(tert-butoxy)carbonyl]amino}ethyl)-3,5-dimethyl-1Hpyrazole-4-carboxylate (247) (1 g, 2.58 mmol, 80% purity) and LiOH (0.37 g, 15.49 mmol) in THF (60 ml) and water (30 ml) at 70°C gave the title compound (0.417 g, 52%, 90% pure) as yellow solid.

1H NMR (DMSO-66, 500 MHz): δ [ppm] = 12.02 (s, 1H), 6.91 (t, J=5.6 Hz, 1H), 4.00 (t, J=6.0Hz, 2H), 3.21(kv, J=6.0Hz, 2H), 2.40(s, 3H), 2.26(s, 3H), 1.37(s, 9H).

HPLC-MS (method A): [m/z]: 284.05 [M+H] ⁺ .

1-(2-{[(tert-Butoxy)carbonyl]amino}ethyl)-3-(trifluoromethyl)-1H-pyrazole-4-carboxylic acid (251)

Similar to general procedure 5, LiOH (0.241 g, 10.08 mmol) and ethyl 1-(2-{[(tert-butoxy)carbonyl]amino}ethyl)-3-(trifluoromethyl)-1H-pyrazole-4-carboxylate (248) (89%, 1.18 g, 2.99 mmol) in THF/water (25 ml/8 ml) at room temperature for 16 h gave the title compound (0.54 g, 45%) as white solid.

1H NMR (CDCl ₃ , 250 MHz): δ [ppm] = 8.03 (s, 1H), 4.80 (br.s, 1H), 4.36-4.18 (br.m, 2H), 3.713.54 (m, 2H), 1.45 (s, 9H).

HPLC-MS (method A): [m/z]: 346.1 [M+Na] ⁺ .

tert-Butyl N-(2-{4-[(pyridin-2-ylmethyl)carbamoyl]-1H-pyrazol-1-yl}ethyl)carbamate (252)

Similarly, using the general procedure 6, 1-(2-{[(tert-butoxy)carbonyl]amino}ethyl)1H-pyrazole-4-carboxylic acid (249) (0.50 g, 1.96 mmol), 1 -(pyridin-2-yl)methanamine (0.30 ml, 2.94 mmol), DIPEA (1.0 ml, 5.88 mmol) and HATU (1.12 g, 2.94 mmol) in DCM (30 mL) gave the title compound (0.647 g, 56%, 59% pure) as a yellow oil after column purification.

- 184 039916 flash chromatography (elution gradient 3:2 EtOAc/heptane and 100% EtOAc). The compound was used in the next step without further purification.

¹ H NMR (CDC13, 500 MHz): δ [ppm] = 8.56 (d, J=4.4 Hz, 1H), 7.87 (d, J=3.8 Hz, 2H), 7.68 (td, J=7.7, 1.8

Hz, 1H), 7.31 (d, J=7.8 Hz, 1H), 7.24-7.20 (m, 1H), 7.14 (s, 1H), 4.84 (s, 1H ), 4.71 (d, J=4.8 Hz, 2H), 4.25 (t,

J=5.4Hz, 2H), 3.58(q, J=5.4Hz, 2H), 1.43(s, 9H).

HPLC-MS (method A): [m/z]: 346.3 [M+H]+.

tert-Butyl N-{2-[4-({5H,6H,7H-cyclopenta[b]pyridin-7-yl}carbamoyl)-1H-pyrazol-1yl]ethyl}carbamate (253)

I—\ ° // ud Ν^\

P λ ⁿ Itz x—NHBoc \^NN

Similarly, using the general procedure 6, 1-(2-{[(tert-Butoxy)carbonyl]amino}ethyl)III-pyrazole-4-carboxylic acid (249) (0.7 g, 2.74 mmol), 5H ,6H,7H-cyclopenta[b]pyridine-7-amine hydrochloride (0.7 g, 4.11 mmol), DIPEA (1.4 ml, 8.23 mmol) and HATU (1.56 g, 4.11 mmol) in DMF (15 mL) gave the title compound (0.709 g, 70%) as an off-white solid after purification by flash column chromatography (80-100% EtOAc/heptane gradient elution).

¹ H NMR (MeOD-d ₆ , 500 MHz): δ [ppm] = 8.35 (d, J=4.6 Hz, 1H), 8.07 (s, 1H), 7.94 ( s, 1H), 7.73 (d, J=7.8 Hz, 1H), 7.28 (dd, J=7.6, 5.0 Hz, 1H), 5.53 (t, J=8 .1 Hz, 1H), 4.23 (t, J=5.7 Hz, 2H), 3.49-3.43 (m, 2H), 3.133.03 (m, 1H), 2.95 (dt , J=16.8, 8.5 Hz, 1H), 2.66 (ddt, J=11.3, 8.1, 3.9 Hz, 1H), 2.07-1.95 (m, 1H ), 1.40 (s, 9H).

HPLC-MS (method A): [m/z]: 372.1 [M+H] ⁺ .

tert-Butyl N-[2-(4-{[(3-fluoropyridin-2-yl)methyl]carbamoyl}-1H-pyrazol-1-yl)ethyl]carbamate (254) F Ϊ ^H A ^ ^NHBoc

Similarly, using the general procedure 6, 1-(2-{[(tert-butoxy)carbonyl]amino}ethyl)1H-pyrazole-4-carboxylic acid (249) (0.35 g, 1.3 mmol), ( 3-fluoropyridin-2-yl)methanamine dihydrochloride (A2) (0.46 g, 1.95 mmol), DIPEA (1.13 ml, 6.51 mmol) and HATU (0.74 g, 1.95 mmol) in DMF (7 mL) gave the title compound (0.4 g, 82%) as an orange foam after purification by flash column chromatography (eluting with a 0-50% MeOH/EtOAc gradient).

¹ H NMR (CDC13, 500 MHz): δ [ppm] = 8.40-8.37 (m, 1H), 7.89 (s, 2H), 7.46-7.38 (m, 1H), 7.30-7.24 (m, 1H), 4.86 (brs, 1H), 4.78 (dd, J=4.4 and 1.2 Hz, 2H), 4.26 (t, J=5.3 Hz, 2H), 3.64-3.50 (m, 2H), 1.43 (s, 9H).

HPLC-MS (method A): [m/z]: 364.40 [M+H] ⁺ .

tert-Butyl N-[2-(4-{[(3-chloropyridin-2-yl)methyl]carbamoyl}-1Н-pyrazol-1-yl)ethyl]carbamate (255) α ?

^nA ^ ^NHBoc

Similarly, using the general procedure 6, 1-(2-{[(tert-butoxy)carbonyl]amino}ethyl)1H-pyrazole-4-carboxylic acid (249) (0.7 g, 2.74 mmol), ( 3-chloropyridin-2-yl)methanamine dihydrochloride (0.89 g, 4.11 mmol), DIPEA (2.39 ml, 13.71 mmol) and HATU (1.56 g, 4.11 mmol) in DMF ( 15 ml) at room temperature for 24 hours gave the title compound (0.79 g, 76%) as a viscous brown oil after purification by flash column chromatography (eluting with a 0-50% MeOH/EtOAc gradient). The resulting oil solidifies on standing.

¹ H NMR (CDC13, 500 MHz): δ [ppm] = 8.48 (dd, J=4.7, 1.0 Hz, 1H), 7.91 (s, 1H), 7.91 (s, 1H), 7.72 (dd, J=8.0, 1.2 Hz, 1H), 7.50 (broad s, 1H), 7.23 (dd, J=8.0, 4 .8 Hz, 1H), 4.87 (broad s, 1H), 4.78 (d, J=4.2 Hz, 2H), 4.26 (t, J=5.3 Hz, 2H), 3.59 (pseudo q, J=5.6 Hz, 2H), 1.44 (s, 9H).

HPLC-MS (method A): [m/z]: 380.4 [M+H] ⁺ .

tert-Butyl N-[2-(4-{[(3-fluoropyridin-2-yl)methyl]carbamoyl}-3,5-dimethyl-1H-pyrazol-1yl)ethyl]carbamate (256)

Di ^Н >N ™°С

Similar to general procedure 6, 1-(2-{[(tert-butoxy)carbonyl]amino}ethyl)-3,5-dimethyl-1Hpyrazole-4-carboxylic acid (250) (0.42 g, 1.33 mmol, 90% pure), (3-fluoropyridin-2-yl)methanamine dihydrochloride (A2) (0.47 g, 2.0 mmol), DIPEA (1.16 ml, 6.67 mmol) and HATU (0.76 g , 2.0 mmol) in THF (15 mL) and DMF (3 mL) gave the title compound (0.40 g, 77%) as a colorless glassy solid after purification by flash column chromatography (kp-NH, eluting with a gradient of 60-100% EtOAc/heptane).

¹ H NMR (DMSO-a ₆ , 500 MHz): δ [ppm] = 8.38 (d, J=4.7 Hz, 1H), 7.81-7.77 (m, 1H), 7.71-7.65 (m, 1H), 7.39 (dt, J=8.6, 4.4 Hz, 1H), 6.94-6.89 (m, 1H), 4.59 ( d, J=4.3 Hz, 2H), 3.99 (t, J=6.5 Hz, 2H), 3.20 (kv, J=6.5

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Hz, 2H), 2.34 (s, 3H), 2.25 (s, 3H), 1.37 (s, 9H).

HPLC-MS (method A): [m/z]: 392.1 [M+H] ⁺ .

tert-Butyl N-[2-(4-{[(3-fluoropyridin-2-yl)methyl]carbamoyl}-3 -(trifluoromethyl)-1 H-pyrazol-1 yl)ethyl]carbamate (257)

Similarly, using General Procedure 6, 1-(2-{[(tert-butoxy)carbonyl]amino}ethyl)-3(trifluoromethyl)-1H-pyrazole-4-carboxylic acid (251) (90%, 0.54 g, 1.53 mmol), (3-fluoropyridin-2yl)methanamine dihydrochloride (A2) (0.23 g, 1.8 mmol), DIPEA (0.87 ml, 5.01 mmol) and HATU (0.57 g, 1.5 mmol) in DMF (3 ml) and THF (15 ml) at room temperature for 72 h gave the title compound (0.646 g, 65%) after purification by flash column chromatography (eluting with a gradient of 5 -100% EtOAc/heptane).

1H NMR (CDCl ₃ , 250 MHz): δ [ppm] = 8.40-8.35 (m, 1H), 7.98 (s, 1H), 7.56 (s, 1H), 7 .46-7.37 (m, 1H), 7.31-7.23 (m, 4H), 4.80 (dd, J=4.3, 1.2 Hz, 2H), 4.30 (t , J=5.4 Hz, 2H), 3.60 (pseudo q, J=6.0 Hz, 2H), 1.44 (s, 9H).

HPLC-MS (Method A): [m/z]: 432.1 [M+H] ⁺ .

1-(2-Aminoethyl)-N-(pyridin-2-ylmethyl)-1H-pyrazole-4-carboxyamide dihydrochloride (258)

Similarly, using general procedure 2, 12 M HC (6.5 mL) and tert-butyl N-(2-{4[(pyridin-2-ylmethyl)carbamoyl]-1H-pyrazol-1-yl}ethyl)carbamate (252) (0.647 g, 1.11 mmol, 59% pure) in MeOH (6.5 mL) at room temperature for 2.5 h gave the title compound (0.513 g, quant) as a pale yellow oils. The compound was used in the next step without further purification.

1H NMR (DMSO-d ₆ , 500 MHz): δ [ppm] = 9.16 (s, 1H), 8.77 (s, 1H), 8.33 (s, 1H), 8.13 (s, 2H), 8.05 (s, 1H), 7.80 (s, 2H), 4.72 (s, 2H), 4.42 (s, 2H), 3.27 (d, J= 5.9 Hz, 2H), 3.05-2.78 (m, 3H).

HPLC-MS (method A): [m/z]: 246 [M+H] ⁺ .

1-(2-Aminoethyl)-N-{5Н,6Н,7Н-cyclopenta[b]pyridin-7-yl}-1Н-pyrazol-4-carboxamide dihydrochloride (259)

Similarly, using the general procedure 2, 12 M HC (3.2 ml) and tert-butyl N-{2-[4({5H,6H,7H-cyclopenta[b]pyridin-7-yl}carbamoyl)-1H -pyrazole-1-sphthyl}carbamate (253) (0.709 g, 1.909 mmol) in MeOH (20 mL) at room temperature over 20 hours gave the title compound (0.745 g, quant) as a gray solid.

1H NMR (500 MHz, Methanol-d ₄ ): δ [ppm] = 8.60 (d, J=5.8 Hz, 1H), 8.51 (d, J=7.8 Hz, 1H ), 8.29 (s, 1H), 8.05 (s, 1H), 7.93 (dd, J=7.7, 6.0 Hz, 1H), 5.80 (t, J=8, 4 Hz, 1H), 4.53-4.49 (m, 2H), 3.46 (t, J=5.7 Hz, 2H), 3.39-3.35 (m, 1H), 3, 18 (dt, J=17.0, 8.5 Hz, 1H), 2.83-2.74 (m, 1H), 2.41-2.31 (m, 1H).

HPLC-MS (Method A): [m/z]: 272.05 [M+H]+.

1-(2-Aminoethyl)-N-[(3-fluoropyridin-2-yl)methyl]-1H-pyrazole-4-carboxamide dihydrochloride (260)

Similarly, using the general procedure 2, 12 M HC (3.67 ml) and tert-butyl N-[2-(4-{[(3fluoropyridin-2-yl)methyl]carbamoyl}-1H-pyrazole-1-shftyl ]carbamate (254) (0.4 g, 1.1 mmol) in MeOH (5 mL) at room temperature over 16 h gave the title compound (0.38 g, quant) as a creamy foam. in the next step without further purification.

1H NMR (DMSO-d ₆ , 500 MHz): δ [ppm] = 8.65 (t, J=5.7 Hz, 1H), 8.38 (dt, J=4.6, 1, 4 Hz, 1H), 8.25 (s, 1H), 8.12 (br s, 3H), 8.00-7.97 (m, 1H), 7.73-7.68 (m, 1H ), 7.43-7.39 (m, 1H), 4.59 (dd, J=5.6, 1.5 Hz, 2H), 4.40 (t, J=6.1 Hz, 2H) , 3.25 (pseudo hept, J=5.7 Hz, 2H).

HPLC-MS (method A): [m/z]: 264.05 [M+H] ⁺ .

-(2-Aminoethyl)-N -[(3-chloropyridin-2-yl)methyl]-1 H-pyrazole-4-carboxamide dihydrochloride (261)

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Similarly, using the general procedure 2, 12 M HC (6.5 ml) and tert-butyl N-[2-(4-{[(3chloropyridin-2-yl)methyl]carbamoyl}-1H-pyrazol-1-yl )ethyl]carbamate (255) (0.79 g, 1.97 mmol) in MeOH (10 mL) at room temperature over 25 h gave the title compound (0.7 g, 95%) as a beige solid . The compound was used in the next step without further purification.

¹ H NMR (Methanol-b ₄ , 500 MHz): δ [ppm] = 8.69 (dd, J=5.5, 1.2 Hz, 1H), 8.51 (dd, J=8 .3, 1.2 Hz, 1H), 8.26 (s, 1H), 8.05 (s, 1H), 7.86 (dd, J=8.3, 5.5 Hz, 1H), 4 .88 (s, 2H), 4.51 (t, J=5.6 Hz, 2H), 3.46 (t, J=5.6 Hz, 2H).

HPLC-MS (method A): [m/z]: 279.9 [M+H] ⁺ .

1-(2-Aminoethyl)-N-[(3-fluoropyridin-2-yl)methyl]-3,5-dimethyl-1Н-pyrazole-4-carboxamide dihydrochloride (262)

Similar to general procedure 2, tert-butyl N[2-(4-{[(3-fluoropyridin-2-yl)methyl]carbamoyl}-3,5dimethyl-1H-pyrazol-1-yl)ethyl]carbamate (256) ( 0.4 g, 1.02 mmol) and 12 M HC (1.7 ml) in MeOH (5 ml) gave the title compound (0.399 g, 93% pure) as a white solid.

¹ H NMR (Methanol-b ₄ , 500 MHz): δ [ppm] = 8.64 (dd, J=5.5, 1.2 Hz, 1H), 8.33 (t, J=8 .9 Hz, 1H), 7.97-7.92 (m, 1H), 4.86-4.85 (m, 2H), 4.34-4.29 (m, 2H), 3.40 ( t, J=5.8 Hz, 2H), 2.46 (s, 3H), 2.38 (s, 3H).

HPLC-MS (method A): [m/z]: 292 [M+H]+.

1-(2-Aminoethyl)-N-[(3-fluoropyridin-2-yl)methyl]-3-(trifluoromethyl)-1H-pyrazole-4-carboxamide dihydrochloride (263)

Similarly, using the general procedure 2, 12 M HC (2.1 ml) and tert-butyl N-[2-(4-{[(3fluoropyridin-2-yl)methyl]carbamoyl}-3-(trifluoromethyl)-1H -pyrazol-1-yl)ethyl]carbamate (257) (80%, 0.68 g, 1.26 mmol) in MeOH (10 mL) at room temperature over 17 h gave the title compound (0.588 g, 93 .4%) as a pink solid. The compound was used in the next step without further purification.

¹ H NMR (DMSO-66, 250 MHz): δ [ppm] = 8.80 (t, J=5.7 Hz, 1H), 8.52 (s, 1H), 8.42-8 .36 (m, 1H), 8.23 (s, 3H), 7.78-7.68 (m, 1H), 7.48-7.39 (m, 1H), 4.59 (dd, J =5.5, 1.3 Hz, 2H), 4.50 (t, J=6.0 Hz, 2H), 3.29 (kv, J=5.7 Hz, 2H), 2.89 (s , 1H), 2.69 (s, 7H).

HPLC-MS (method A): [m/z]: 332.2 [M+H] ⁺ .

1-{2-[(1H-1,3-Benzodiazol-2-ylmethyl)amino]ethyl}-N-(pyridin-2-ylmethyl)-1H-pyrazol-4carboxamide (illustrative example No. 92)

Similarly, using General Procedure 3, 1-(2-aminoethyl)-N-(pyridin-2-ylmethyl)-1Hpyrazol-4-carboxamide dihydrochloride (258) (256 mg, 0.805 mmol), 1H-benzimidazol-2-carbaldehyde (129 mg, 0.885 mmol) and DIPEA (0.70 ml, 4.02 mmol) in MeOH (7 ml) at room temperature for 65 h, followed by addition of NaBH ₄ (46 mg, 1.21 mmol), gave the title compound (58 mg, 19%) as a pale yellow solid after purification by preparative HPLC.

1H NMR (DMSO-b ₆ , 500 MHz): δ [ppm] = 12.17 (s, 1H), 8.68 (t, J=6.0 Hz, 1H), 8.49 (d , J=4.2 Hz, 1H), 8.23 (s, 1H), 7.91 (s, 1H), 7.74 (td, J=7.7, 1.8 Hz, 1H), 7 .51 (s, 1H), 7.45 (s, 1H), 7.29 (d, J=7.8 Hz, 1H), 7.25 (dd, J=7.1, 5.1 Hz, 1H), 7.18-7.05 (m, 2H), 4.49 (d, J=6.0 Hz, 2H), 4.22 (t, J=6.2 Hz, 2H), 3, 91 (s, 2H), 2.97 (t, J=6.1 Hz, 2H), 2.45 (s, 1H).

HPLC-MS (method B): [m/z]: 376.2 [M+H] ⁺ .

1-{2-[(1H-1,3-Benzodiazol-2-ylmethyl)amino]ethyl}-N-{5H,6H,7H-cyclopenta[b]pyridin-7-yl}-1Hpyrazol-4-carboxamide ( illustrative example No. 132)

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Similarly, using the general procedure 3, 1-(2-Aminoethyl)-N-{5H,6H,7Hcyclopenta[b]pyridin-7-yl}-1H-pyrazole-4-carboxamide dihydrochloride (259) (150 mg, 0 .38 mmol), 1Hbenzimidazole-2-carbaldehyde (67 mg, 0.46 mmol) and DIPEA (0.267 ml, 1.53 mmol) in MeOH (6 ml) at room temperature for 24 h, followed by the addition of NaBH ₄ ( 22 mg, 0.57 mmol) gave the title compound (23 mg, 15%) as a yellow solid after purification by basic prep-HPLC, followed by flash column chromatography (kp-NH, eluting with a 0- 20% MeOH/EtOAc).

¹ H NMR (Methanol-d ₄ , 500 MHz): δ [ppm] = 8.35 (d, J=4.9 Hz, 1H), 8.17 (s, 1H), 7.93 ( s, 1H), 7.74 (d, J=7.6 Hz, 1H), 7.51 (s, 2H), 7.28 (dd, J=7.6, 4.9 Hz, 1H), 7.20 (dd, J=6.0, 3.2 Hz, 2H), 5.54 (t, J=8.2 Hz, 1H), 4.29 (t, J=5.9 Hz, 2H ), 4.01 (s, 2H), 3.09 (t, J=5.9 Hz, 2H), 3.08-3.04 (m, 1H), 2.99-2.91 (m, 1H), 2.71-2.63 (m, 1H), 2.06-1.97 (m, 1H).

HPLC-MS (method B): [m/z]: 402.1 [M+H]+.

1-{2-[(1H-1,3-Benzodiazol-2-ylmethyl)amino]ethyl}-N-[(3-fluoropyridin-2-yl)methyl]-1H-niprazol-4carboxamide (Illustrative Example No. 145)

Similarly, using General Procedure 3, 1-(2-aminoethyl)-N-[(3-fluoropyridin-2yl)methyl]-1H-pyrazole-4-carboxamide dihydrochloride (260) (200 mg, 0.59 mmol), 1H-benzimidazole-2carbaldehyde (104 mg, 0.71 mmol) and DIPEA (0.41 ml, 2.38 mmol) in MeOH (8 ml) at room temperature for 18 h, followed by addition of NaBH ₄ (32.7 mg, 0.87 mmol) gave the title compound (91 mg, 40%) as a colorless film after purification by flash column chromatography (kp-NH, 0-10% MeOH/DCM gradient elution), followed by basic prepHPLC .

1H NMR (DMSO-66, 500 MHz): δ [ppm] = 12.15 (brs, 1H), 8.53 (t, J=5.7 Hz, 1H), 8.37 ( dt, J=4.6, 1.4 Hz, 1H), 8.21 (s, 1H), 7.88 (s, 1H), 7.70-7.65 (m, 1H), 7.59 -7.41 (m, 2H), 7.41-7.37 (m, 1H), 7.12 (d, J=4.7 Hz, 2H), 4.57 (dd, J=5.7 , 1.6 Hz, 2H), 4.21 (t, J=6.2 Hz, 2H), 3.90 (brs, 2H), 2.96 (pseudo brs, 2H).

HPLC-MS (method C): [m/z]: 394.1 [M+H] ⁺ .

1-{2-[(1H-1,3-Benzodiazol-2-ylmethyl)amino]ethyl }-N-[(3-chloropyridin-2-yl)methyl]-1H-pyrazole-4carboxamide (Illustrative Example No. 138 )

Similarly, using General Procedure 3, 1-(2-aminoethyl)-N-[(3-chloropyridin-2yl)methyl]-1H-pyrazole-4-carboxamide dihydrochloride (261) (200 mg, 0.54 mmol), 1H-benzimidazole-2carbaldehyde (94.5 mg, 0.65 mmol) and DIPEA (0.38 ml, 2.16 mmol) in MeOH (8 ml) at room temperature for 16 h, followed by the addition of NaBH ₄ (30 .6 mg, 0.81 mmol) gave the title compound (127 mg, 56%) as a light green foam after purification by basic pre-HPLC.

1H NMR (DMSO-66, 500 MHz): δ [ppm] = 12.17 (s, 1H), 8.51-8.46 (m, 2H), 8.23 (s, 1H), 7.92 (dd, J=8.1, 1.4 Hz, 1H), 7.90 (s, 1H), 7.58-7.41 (breadth m, 2H), 7.37 (dd, J=8.1, 4.7 Hz, 1H), 7.17-7.08 (m, 2H), 4.62 (d, J=5.6 Hz, 2H), 4.23 (t, J =6.2 Hz, 2H), 3.92 (brs, 2H), 2.98 (brs, 2H).

HPLC-MS (method C): [m/z]: 410.2 [M+H] ⁺ .

1-{2-[(1H-1,3-Benzodiazol-2-ylmethyl)amino]ethyl}-N-[(3-fluoropyridin-2-yl)methyl]-3,5-dimethyl1H-pyrazole-4-carboxamide (illustrative example #142)

Similarly, using General Procedure 3, 1-(2-aminoethyl)-N-[(3-fluoropyridin-2yl)methyl]-3,5-dimethyl-1H-pyrazole-4-carboxamide dihydrochloride (262) (150 mg, 0.384 mmol), 1H-188 039916 benzimidazole-2-carbaldehyde (73 mg, 0.499 mmol), DIPEA (0.268 ml, 1.537 mmol) and MgSO ₄ (100 mg) in

MeOH (7 ml) at room temperature for 16 hours, followed by the addition of NaBH ₄ (22 mg,

0.576 mmol) gave the title compound (14 mg, 9%) as a yellow solid after purification by basic prep-HPLC.

1H NMR (DMSO-d6, 500 MHz): δ [ppm] = 12.16 (s, 1H), 8.40-8.37 (m, 1H), 7.84 (t, J=5 .6 Hz, 1H), 7.727.66 (m, 1H), 7.56-7.50 (m, 1H), 7.46-7.42 (m, 1H), 7.40 (dt, J= 8.4, 4.4 Hz, 1H), 7.17-7.09 (m, 2H), 4.59 (d, J=4.3 Hz, 2H), 4.05 (t, J=6 .4 Hz, 2H), 3.92 (s, 2H), 2.90 (t, J=6.4 Hz, 2H), 2.38 (s, 3H), 2.24 (s, 3H).

HPLC-MS (Method D): [m/z]: 422.3 [M+H]+.

1-{2-[(1H-1,3-Benzodiazol-2-ylmethyl)amino]ethyl}-N-[(3-fluoropyridin-2-yl)methyl]-3(trifluoromethyl)-1H-pyrazol-4- carboxamide (illustrative example No. 156)

Similarly, using General Procedure 3, 1-(2-aminoethyl)-N-[(3-fluoropyridin-2yl)methyl]-3-(trifluoromethyl)-1H-pyrazole-4-carboxamide dihydrochloride (263) (81%, 100 mg, 0.20 mmol), N-benzimidazole-2-carbaldehyde (35 mg, 0.24 mmol) and DIPEA (0.14 ml, 0.8 mmol) in MeOH (5 ml) at room temperature for 16 h followed by addition of NaBH ₄ (22 mg, 0.6 mmol) gave the title compound (12 mg, 13%) as a white solid after purification by basic prep-HPLC followed by flash column chromatography (elution in a 010% MeOH/DCM gradient).

1H NMR (DMSO-H6, 500 MHz): δ [ppm] = 12.15 (s, 1H), 8.69 (t, 1H), 8.45 (s, 1H), 8.37 ( d, 1H), 7.72-7.66 (m, 1H), 7.53 (d, 1H), 7.45-7.38 (m, 2H), 7.12 (t, 2H), 4 .71-4.45 (m, 2H), 4.29 (t, 2H), 3.92 (s, 2H), 2.99 (s, 2H).

HPLC-MS (Method C): [m/z]: 462.1 [M+H]+.

-(2-{[2-(1H-1,3-Benzodiazol-2-yl)ethyl]amino}ethyl)-N-(pyridin-2-ylmethyl)-1H-pyrazole-4carboxamide (Illustrative Example No. 109) o ⁴ ©N ^Η ϋ

Similarly, using the general procedure 7, 1-(2-aminoethyl)-N-(pyridin-2-ylmethyl)-1Hpyrazol-4-carboxamide dihydrochloride (258) (250 mg, 0.79 mmol), 2-(2- chloroethyl)-1H-benzimidazole hydrochloride (340 mg, 1.58 mmol) and DIPEA (2.1 ml, 11.8 mmol) in DMF (7.5 ml) at room temperature for 6 days gave the title compound ( 17 mg, 5%) as a pale yellow solid after purification by preparative HPLC, followed by (KP-NH) flash column chromatography using 4:6 EtOAc/heptane to 100% EtOAc, then 20% MeOH: 80% DCM , then prep-HPLC (MeCN/water, 2 mM NH4CO3).

1H NMR (DMSO-66, 500 MHz): δ [ppm] = 12.10 (s, 1H), 8.68 (t, J=6.0 Hz, 1H), 8.50 (d, J=4.1 Hz, 1H), 8.21 (s, 1H), 7.90 (s, 1H), 7.74 (td, J=7.7, 1.8 Hz, 1H), 7, 58-7.33 (m, 2H), 7.29 (d, J=7.9 Hz, 1H), 7.25 (dd, J=7.4, 4.9 Hz, 1H), 7.09 (dd, J=6.0, 2.8 Hz, 2H), 4.49 (d, J=6.0 Hz, 2H), 4.19 (t, J=6.2 Hz, 2H), 3 .00-2.93 (m, 4H), 2.91 (t, J=6.2 Hz, 2H), 2.05 (s, 1H).

HPLC-MS (method B): [m/z]: 390.2 [M+H] ⁺ .

-(2-{[2-(1H-1,3-Benzodiazol-2-yl)ethyl]amino}ethyl)-N-[(3-fluoropyridin-2-yl)methyl]-1H-pyrazole4-carboxamide tetrahydrochloride (illustrative example No. 111)

Similarly, using the general procedure 7, 1-(2-aminoethyl)-N-[(3-fluoropyridin-2yl)methyl]-1H-pyrazole-4-carboxamide dihydrochloride (260) (0.7 g, 2.08 mmol ), 2-(2-chloroethyl)-1Hbenzimidazole (0.451 g, 2.499 mmol) and DIPEA (5.4 ml, 31.23 mmol) in DMF (15 ml) at 30° C. for 6 days gave the product as free base (63 mg) after purification by silica gel column chromatography (kp-NH, elution with 0-40% MeOH/EtOAc gradient), followed by basic prep-HPLC. The free base product was dissolved in MeOH (3 ml) and 12 M HC (2 ml) at room temperature for 2 hours to give the title compound (65 mg, 5.6%) as a brown solid. .

1H NMR (Methanol-b ₄ , 500 MHz): δ [ppm] = 8.46 (s, 1H), 8.25 (s, 1H), 8.01 (s, 1H), 7.81 (dd, J=6.2, 3.2 Hz, 3H), 7.63 (dd, J=6.2, 3.1 Hz, 3H), 4.78 (s, 2H), 4.64 ( s, 2H), 3.72 (d, J=7.6 Hz, 6H).

HPLC-MS (method B): [m/z]: 408.2 [M+H] ⁺ .

1-(2-{[2-(1H-1,3-Benzodiazol-2-yl)ethyl]amino}ethyl)-N-{5H,6H,7H-cyclopenta[b]pyridin-7-yl}1H- pyrazole-4-carboxamide trihydrochloride (illustrative example No. 131)

Similarly, using the general procedure 7, 1-(2-Aminoethyl)-N-{5H,6H,7Hcyclopenta[b]pyridin-7-yl}-1H-pyrazole-4-carboxamide dihydrochloride (259) (0.2 g , 0.51 mmol), 2-(2chloroethyl)-1H-benzimidazole hydrochloride (0.13 g, 0.61 mmol) and DIPEA (0.44 ml, 2.55 mmol) in DMF (3 ml) at room temperature for 72 h, then at 40°C for 21 h gave the product in the form of a free base (55 mg) after purification by basic prep-HPLC. The free base product was then dissolved in MeOH (3 ml) and 12M HC (0.5 ml) at room temperature for 2 hours to give the title compound (0.049 g, 18%) as a brown solid.

1H NMR (Methanol-b ₄ , 500 MHz): δ [ppm] = 8.60 (d, J=5.2 Hz, 1H), 8.51 (d, J=6.8 Hz, 1H ), 8.31 (s, 1H), 8.03 (s, 1H), 7.95-7.91 (m, 1H), 7.83-7.79 (m, 2H), 7.65- 7.62 (m, 2H), 5.80 (t, J = 8.5 Hz, 1H), 4.68-4.64 (m, 2H), 3.77-3.70 (m, 6H) , 3.34 (d, J=3.3 Hz, 1H), 3.23-3.17 (m, 1H), 2.83-2.75 (m, 1H), 2.39-2.30 (m, 1H).

HPLC-MS (method B): [m/z]: 416.1 [M+H]+.

1-(2-{[2-(1H-1,3-Benzodiazol-2-yl)ethyl]amino}ethyl)-N-[(3-chloropyridin-2-yl)methyl]-1H-pyrazole-4carboxamide (illustrative example #144)

Similarly, using the general procedure 7, 1-(2-Aminoethyl)-N-[(3-chloropyridin-2yl)methyl]-1H-pyrazole-4-carboxamide dihydrochloride (261) (0.45 g, 1.28 mmol ), 2-(2-chloroethyl)-1Hbenzimidazole (0.28 g, 1.53 mmol) and DIPEA (3.3 ml, 19.14 mmol) in DMF (7 ml) at room temperature for 66 h, then at 40°C for 90 h, gave the title compound (112 mg, 21%) as a colorless film after purification by flash column chromatography (kp-NH, 0-10% MeOH/DCM gradient elution), s followed by basic prep-HPLC.

¹ H NMR (DMSO-06, 500 MHz): δ [ppm] = 12.10 (s, 1H), 8.52-8.44 (m, 2H), 8.22 (s, 1H) , 7.92 (dd, J=8.1, 1.4 Hz, 1H), 7.90 (s, 1H), 7.55-7.38 (breadth m, 2H), 7.37 (dd , J=8.1, 4.7 Hz, 1H), 7.13-7.07 (m, 2H), 4.63 (d, J=5.7 Hz, 2H), 4.20 (t, J=6.2 Hz, 2H), 2.99-2.93 (m, 4H), 2.93-2.89 (m, 2H).

HPLC-MS (method C): [m/z]: 424.1 [M+H]+.

1-(2-{[2-(1H-1,3-Benzodiazol-2-yl)ethyl]amino}ethyl)-N-[(3-fluoropyridin-2-yl)methyl]-3(trifluoromethyl)-1H -pyrazole-4-carboxamide (illustrative example No. 162)

Similarly, using General Method 8, DBU (62.94 µl, 0.42 mmol),

1-(2-aminoethyl)-N-[(3-fluoropyridin-2-yl)methyl]-3-(trifluoromethyl)-1H-pyrazole-4-carboxamide dihydrochloride (263) (81%, 140 mg, 0.28 mmol) and N-(2-nitrophenyl)prop-2-enamide (D) (52.84 mg, 0.27 mmol) in MeCN (5 ml) at room temperature over 16 hours gave a residue (0.160 g) as mixtures of mono:bis-alkylated (1:1) adducts.

The resulting residue was dissolved in AcOH (4 ml). Iron (0.03 g) was added and the reaction mixture was heated at 80° C. for 1 hour. The reaction mixture was cooled and diluted with water (5 ml). The mixture was cooled in an ice bath and basified to pH 12 with 10M NaOH solution. Water (10 ml) was added and the resulting mixture was extracted with CHC ₃ :IPA (4:1, 4x30 ml). The combined organic layers were dried (Na ₂ SO ₄ ), filtered and evaporated. Purification by flash column chromatography (gradient elution with 5-10% MeOH/DCM followed by 0-10% 7 N NH ₃ in MeOH/DCM), followed by basic prep-HPLC purification, gave the title compound (19 mg, 13.9%) as a colorless oil.

¹ H NMR (DMSO-d ₆ , 500 MHz): δ [ppm] = 12.11 (broad s, 1H), 8.69 (t, J=5.6 Hz, 1H), 8, 43 (s, 1H), 8.428.35 (m, 1H), 7.74-7.65 (m, 1H), 7.52-7.36 (bdm, 2H), 7.43-7, 37 (m, 1H), 7.15-7.06 (breadth m, 2H), 4.59-4.55 (m, 2H), 4.26 (t, J=6.1 Hz, 2H) , 3.02-2.94 (m, 4H), 2.94-2.89 (m, 2H).

HPLC-MS (Method C): [m/z]: 476.2 [M+H] ⁺ .

Ethyl 1-[2-(benzyloxy)-2-oxoethyl]-1Н-pyrazole-4-carboxylate (268)

Benzyl bromoacetate (2.82 ml, 18 mmol) was added dropwise to an ice-cold suspension of ethyl 1Hpyrazole-4-carboxylate (2.3 g, 16 mmol) and K2CO3 (3.4 g, 25 mmol) in DMF (20 ml). The resulting mixture was warmed to room temperature and stirred for 2 hours. The resulting mixture was diluted with diethyl ether (100 ml) and washed with water (2x40 ml) and saturated sodium chloride solution (5x20 ml). The organic phase was dried (Na ₂ SO ₄ ), filtered and evaporated in vacuo. Purification by flash column chromatography eluting with a 0-100% DCM/heptane gradient gave the title compound (1.57 g, 33%) as a colorless oil.

1H NMR (CDCl ₃ , 500 MHz): δ [ppm] = 8.01 (s, 1H), 7.98 (s, 1H), 7.44-7.32 (m, 5H), 5 .24 (s, 2H), 4.98 (s, 2H), 4.32 (q, J=7.1 Hz, 2H), 1.37 (t, J=7.1 Hz, 3H).

HPLC-MS (method A): [m/z]: 288.95 [M+H] ⁺ .

2-[4-(Ethoxycarbonyl)-1H-pyrazol-1-yl]acetic acid (269)

A suspension of ethyl 1-[2-(benzyloxy)-2-oxoethyl]-1H-pyrazole-4-carboxylate (268) (1.57 g, 5.44 mmol) and palladium on charcoal (10% wt, 0.590 g, 0 .55 mol) in EtOH (50 ml) was stirred under a hydrogen atmosphere for 2.5 h. The resulting mixture was filtered through a pad of celite and the resulting residue was washed with MeOH. The combined filtrates were evaporated in vacuo to give the title compound (0.99 mg, 92%) as a yellow solid.

1H NMR (CDCl3, 500 MHz): δ [ppm] = 8.07 (s, 1H), 8.03 (s, 1H), 5.07 (s, 2H), 4.34 (q, J=7.1 Hz, 2H), 1.37 (t, J=7.1 Hz, 3H).

HPLC-MS (method A): [m/z]: 198.9 [M+H] ⁺ .

Ethyl 1-({[2-(1H-1,3-benzodiazol-2-yl)ethyl]carbamoyl}methyl)-1H-pyrazole-4-carboxylate (270)

Similar to general procedure 6, HATU (1.1 g, 2.88 mmol), 2-[4-(ethoxycarbonyl)-1H-pyrazol-1yl]acetic acid (269) (0.476 g, 2.4 mmol), 2- (1H-1,3-benzodiazol-2-yl)ethan-1-amine dihydrochloride (0.62 g, 2.64 mmol) and DIPEA (1.67 ml, 9.61 mmol) in THF (30 ml) at room temperature for 2 hours gave the title compound (0.676 g, 82%) as a white solid after purification by flash column chromatography (eluting with a gradient of 0-8% MeOH/DCM).

1H NMR (CDCl ₃ , 500 MHz): δ [ppm] = 8.01 (s, 1H), 7.87 (s, 1H), 7.62 (m, 2H), 7.36 (m , 3H), 4.90 (s, 2H), 4.28 (kv, J=7.1 Hz, 2H), 3.80 (kv, J=5.9 Hz, 3H), 3.39 (t , J=5.9 Hz, 2H), 1.34 (t, J=7.1 Hz, 3H).

HPLC-MS (method A): [m/z]: 342.1 [M+H] ⁺ .

1-({[2-(1H-1,3-Benzodiazol-2-yl)ethyl]carbamoyl}methyl)-1H-pyrazole-4-carboxylic acid (271)

Similar to general procedure 5, ethyl 1-({[2-(1H-1,3-benzodiazol-2-yl)ethyl]carbamoyl}methyl)-1Hpyrazole-4-carboxylate (270) (667 mg, 1.95 mmol) and LiOH (140 mg, 5.85 mmol) in THF/water (30 ml/4 ml) at room temperature for 16 hours gave the crude title compound (420 mg) as a white residue.

HPLC-MS (method A): [m/z]: 314 [M+H]+.

1-({[2-(1H-1,3-Benzodiazol-2-yl)ethyl]carbamoyl}methyl)-N-[(3-fluoropyridin-2-yl)methyl]-1 Npyrazole-4-carboxamide dihydrochloride ( illustrative example No. 117)

- 191 039916

Similar to general procedure 6, crude 1-({[2-(1H-1,3-benzodiazol-2-yl)ethyl]carbamoyl}methyl)-1Hpyrazole-4-carboxylic acid (271) (200 mg), (3- fluoropyridin-2-yl)methanamine dihydrochloride (A2) (140 mg, 0.7 mmol), DIPEA (0.44 ml, 2.55 mmol) and HATU (290 mg, 0.76 mmol) in THF (10 ml) and DMF (2 mL) gave the title compound as the free base after purification by flash column chromatography (0-20% MeOH/DCM gradient elution), followed by basic pre-HPLC. The resulting free base was converted to di-HCl salt (28 mg, 9%) after reaction with 12M HC (1 ml) in MeOH (5 ml).

1H NMR (Methanol-d ₄ , 500 MHz): δ [ppm] = 8.49 (d, J=5.2 Hz, 1H), 8.13 (s, 1H), 8.00-7 .92 (m, 2H), 7.76 (dt, J=6.7, 3.4 Hz, 2H), 7.67 (dt, J=8.8, 4.8 Hz, 1H), 7. 59 (dt, J=6.2, 3.4 Hz, 2H), 4.88 (s, 2H), 4.79 (d, J=1.4 Hz, 2H), 3.77 (t, J =6.4 Hz, 2H), 3.39 (t, J=6.4 Hz, 2H).

HPLC-MS (method C): [m/z]: 422.2 [M+H]+.

The above general scheme 15:

1-{1-[(tert-butoxy)carbonyl]piperidin-4-yl}-1H-pyrazole-4-carboxylic acid (272) o

Ethyl 1H-pyrazole-4-carboxylate (0.2 g, 1.427 mmol), tert-butyl 4-[(methylsulfonyl)oxy]piperidine-1-carboxylate (0.362 g, 1.296 mmol) in DMF (5 ml) was stirred for 5 min . The reaction mixture was cooled to 0° C. and NaH (0.234 g, 5.85 mmol) was added portionwise. After the evolution of gas, the reaction was heated at 50°C for 26 hours, the Reaction mixture was cooled, quenched with water (15 ml) and evaporated to dryness. Water (40 ml) was added and the aqueous layer was extracted with ethyl acetate (3x15 ml). The aqueous layer was acidified with saturated KHSO4 to pH 3 and extracted with ethyl acetate (4x20 ml). The combined organic layers were dried (MgSO ₄ ), filtered and evaporated to give the title compound (0.331 g, 79%) as a pale yellow solid.

¹ H NMR (DMSO-d6, 500 MHz): δ [ppm] = 8.29 (s, 1H), 7.81 (s, 1H), 4.44-4.36 (m, 1H) , 4.08-3.97 (m, 2H), 2.89 (s, 2H), 2.01-1.95 (m, 2H), 1.79 (q.d., J=12.4, 4.4 Hz, 2H), 1.41 (s, 9H).

HPLC-MS (method A): [m/z]: 318.0 [M+Na] ⁺ .

tert-Butyl 4-(4-{[(3-Fluoropyridin-2-yl)methyl]carbamoyl} -1 H-pyrazol-1 -yl)piperidine-1 carboxylate(273)

F O

Similarly, using the general procedure 6, 1-{1-[(tert-butoxy)carbonyl]piperidin-4yl}-1H-pyrazole-4-carboxylic acid (272) (301 mg, 1.02 mmol), 3-fluoropyridine -2-yl)methanamine dihydrochloride (A2) (358 mg, 1.53 mmol), DIPEA (0.8 ml), HATU (581 mg, 1.53 mmol) in THF (15 ml) and DMF (3 ml) gave the title compound (315 mg, 65%) as a glassy solid after purification by flash column chromatography (eluting with a 20-100% EtOAc/heptane gradient).

1H NMR (DMSO-J6, 500 MHz): δ [ppm] = 8.51 (t, J=5.7 Hz, 1H), 8.37 (dt, J=4.6 and 1.5 Hz, 1H), 8.25 (s, 1H), 7.89 (s, 1H), 7.71-7.64 (m, 1H), 7.42-7.36 (m, 1H), 4 .57 (dd, J=5.7, 1.5 Hz, 2H), 4.44-4.32 (m, 1H), 4.043.96 (m, 2H), 2.89 (brs, 2H ), 2.01 (s, 2H), 1.74 (q.d., J=12.4, 4.4 Hz, 2H), 1.41 (s, 9H).

HPLC-MS (method A): [m/z]: 404.1 [M+H] ⁺ .

N-[(3-Fluoropyridin-2-yl)methyl]-1-(piperidin-4-yl)-1H-pyrazol-4-carboxamide dihydrochloride (274)

F O

2HCI

Similarly, using General Procedure 2, 12M HC (1.1 mL) and tert-butyl 4-(4-{[(3fluoropyridin-2-yl)methyl]carbamoyl}-1H-pyrazol-1-yl)piperidin-1 -carboxylate (273) (315 mg, 0.664 mmol) in MeOH (4 mL) at 35° C. for 16 hours gave the title compound (277 mg) as an off-white solid. The compound was used in the next step without further purification.

1H NMR (Methanol-J4, 500 MHz): δ [ppm] = 8.54-8.47 (m, 1H), 8.22 (d, J=2.4 Hz, 1H), 8, 00-8.11 (m, 1H), 7.98 (s, 1H), 7.69-7.78 (m, 1H), 4.80 (s, 2H), 4.66-4.56 ( m, 1H), 3.56 (dt, J=13.1, 3.4 Hz, 2H), 3.22 (td, J=12.8, 3.4 Hz, 2H), 2.38-2 .19 (m, 4H).

- 192 039916

HPLC-MS (method A): [m/z]: 304 [M+H]+.

1-[1-(1H-1,3-Benzodiazol-2-ylmethyl)piperidin-4-yl]-N-[(3-fluoropyridin-2-yl)methyl]-1H-pyrazole4-carboxamide (illustrative example no. 150)

Similar to general procedure 7, 2-(chloromethyl)-1H-benzimidazole (71 mg, 0.424 mmol), N-[(3fluoropyridin-2-yl)methyl]-1-(piperidin-4-yl)-1H-pyrazol-4 -carboxamide (274) (99 mg, 0.326 mmol) and DIPEA (0.171 ml, 0.98 mmol) in DMF (2 ml) at 30° C. for 16 h gave the title compound (21 mg, 15%) in as a white solid after purification by basic prep-HPLC, followed by purification by flash column chromatography (kp-NH, eluting with a 020% MeOH/EtOAc gradient).

¹ H NMR (DMSO-a ₆ , 500 MHz): δ [ppm] = 12.28 (s, 1H), 8.52 (t, J=5.7 Hz, 1H), 8.37 ( dt, J=4.6, 1.5 Hz, 1H), 8.22 (s, 1H), 7.89 (s, 1H), 7.71-7.65 (m, 1H), 7.54 (brs, 1H), 7.44 (brs, 1H), 7.42-7.36 (m, 1H), 7.14 (brs, 2H), 4.58 (dd, J =5.7, 1.6 Hz, 2H), 4.26-4.12 (m, 1H), 3.77 (s, 2H), 2.95 (d, J=12.0 Hz, 2H) , 2.322.23 (m, 2H), 2.05-1.94 (m, 4H).

HPLC-MS (Method D): [m/z]: 434.3 [M+H]+.

1-{1-[2-(1H-1,3-Benzodiazol-2-yl)ethyl]piperidin-4-yl}-N-[(3-fluoropyridin-2-yl)methyl]-1Hpyrazol-4-carboxamide (illustrative example #198)

Similar to general procedure 8, DBU (0.28 mL, 1.89 mmol), N-[(3-fluoropyridin-2-yl)methyl]-1(piperidin-4-yl)-1H-pyrazol-4-carboxamide dihydrochloride (274) (248 mg, 0.47 mmol) and N-(2nitrophenyl)prop-2-enamide (D) (154 mg, 0.8 mmol) in MeCN (10 ml) at room temperature for 17 h gave the target intermediate (200 mg, 82%) after purification by flash column chromatography (elution with 0-25% MeOH/EtOAc gradient) to give the title compound as a yellow solid.

HPLC-MS (method A): [m/z]: 496.1 [M+H]+.

The resulting intermediate was further reacted with iron powder (87 mg, 1.56 mmol) in AcOH (3 mL) at 80°C for 1.5 h to give the title compound (72 mg, 33%) as an off-white solid after purification by flash column chromatography eluting with 0-20% MeOH/DCM.

1H NMR (DMSOCH, 500 MHz): δ [ppm] = 12.14 (s, 1H), 8.51 (t, J=5.7 Hz, 1H), 8.38 (dt, J= 4.6, 1.5 Hz, 1H), 8.24 (s, 1H), 7.89 (s, 1H), 7.72-7.65 (m, 1H), 7.51 (s, 1H ), 7.46-7.37 (m, 2H), 7.11 (d, J=5.1 Hz, 2H), 4.58 (dd, J=5.7, 1.6 Hz, 2H) , 4.22-4.12 (m, 1H), 3.07-2.96 (m, 4H), 2.82 (t, J=7.5 Hz, 2H), 2.23-2.14 (m, 2H), 2.051.97 (m, 2H), 1.95-1.86 (m, 2H).

HPLC-MS (method C): [m/z]: 448.3 [M+H]+.

tert-Butyl 3-{[4-(ethoxycarbonyl)-1 H-pyrazol-1-yl]methyl}piperidine-1-carboxylate (275)

Ethyl 1H-pyrazole-4-carboxylate (334 mg, 2.38 mmol), tert-butyl 3-(hydroxymethyl)piperidine-1-carboxylate (667 mg, 3.1 mmol) and triphenylphosphine (1.25 g, 4.8 mmol ) was dissolved in THF (30 ml) and cooled in an ice bath. DIAD (0.94 ml, 4.8 mmol) was added dropwise. The mixture was stirred while warming to room temperature for 18 hours. The reaction mixture was diluted with ethyl acetate (80 ml) and washed with saturated aqueous NaHCO ₃ (50 ml) and saturated sodium chloride solution (50 ml). The organic phase was dried (Na ₂ SO ₄ ), filtered and evaporated in vacuo. Purification by flash column chromatography eluting with a 0-40% EtOAc/heptane gradient gave the crude title compound (2.23 g) as a colorless oil. The crude product was used in the next step without further purification.

HPLC-MS (method A): [m/z]: 338.15 [M+H]+.

1-({1-[(tert-Butoxy)carbonyl]piperidin-3-yl}methyl)-1H-pyrazole-4-carboxylic acid (276)

- 193 039916

Similarly, using General Procedure 5, tert-Butyl 3-{[4-(ethoxycarbonyl)-1Hpyrazol-1-yl]methyl}piperidine-1-carboxylate (275) (2.23 g, 4.69 mmol, 71% purity) and LiOH (1.12 g, 0.05 mol) in THF/water (20 ml/20 ml) gave the crude product (638 mg, 16%, 38% purity) as a colorless oil.

HPLC-MS (method A): [m/z]: 310.05 [M+H]+.

tert-Butyl 3-[(4-{[(3-fluoropyridin-2-yl)methyl]carbamoyl}-1 H-pyrazol-1-yl)methyl]piperidine-1-carboxylate(277) f o ||Υ% γΑ

Cn ^N U

N )---y / NBoc

Similarly, using the general procedure 6, 1-({1-[(tert-butoxy)carbonyl]piperidin-3yl}methyl)-1H-pyrazole-4-carboxylic acid (276) (0.638 g, 2.06 mmol), (3-fluoropyridin-2yl)methanamine dihydrochloride (A2) (0.451 g, 2.27 mmol), DIPEA (1.26 ml, 7.2 mmol) and HATU (1.18 g, 3.1 mmol) in THF ( 40 mL) and DMF (5 mL) gave the crude title compound (0.348 g, 32%, 79% pure) as a hygroscopic yellow solid after purification by flash column chromatography (elution gradient 0-100% EtOAc/heptane, then 4-8% MeOH/EtOAc).

HPLC-MS (method A): [m/z]: 418.15 [M+H] ⁺ .

N-[(3-Fluoropyridin-2-yl)methyl]-1-(piperidin-3-ylmethyl)-1H-pyrazole-4-carboxamide (278) °

)—\/NH

Similarly, using General Procedure 2, tert-butyl 3-[(4-{[(3-fluoropyridin-2yl)methyl]carbamoyl}-1H-pyrazol-1-yl)methyl]piperidine-1-carboxylate (277) ( 348 mg, 0.83 mmol) and 12 M HC (0.69 mL) gave the title compound (182 mg, 64%) as the free base after purification on an SCX-2 cartridge with DCM and MeOH washes, then eluting with 7N . a solution of ammonia in MeOH.

¹ H NMR (Methanol-d4, 500 MHz): δ [ppm] = 8.36 (dt, J=4.7, 1.1 Hz, 1H), 8.18 (s, 1H), 7 .97 (s, 1H), 7.60 (ddd, J=9.8, 8.4, 1.2 Hz, 1H), 7.39 (dt, J=8.6, 4.4 Hz, 1H ), 4.73 (d, J=1.5 Hz, 2H), 4.18 (td, J=7.1, 2.6 Hz, 2H), 3.29-3.22 (m, 1H) , 3.19-3.13 (m, 1H), 2.81 (td, J=12.5, 3.1 Hz, 1H), 2.70 (t, J=12.1 Hz, 1H), 2.42-2.30(m, 1H), 1.94-1.85(m, 1H), 1.83-1.66(m, 2H), 1.35-1.22(m, 1H ).

HPLC-MS (method A): [m/z]: 318.10 [M+H] ⁺ .

1-{[1-(1H-1,3-Benzodiazol-2-ylmethyl)piperidin-3-yl]methyl}-N-[(3-fluoropyridin-2-yl)methyl]-1Hpyrazol-4-carboxamide (illustrative example no. 153)

Similarly, using General Procedure 7, N-[(3-fluoropyridin-2-yl)methyl]-1(piperidin-3-ylmethyl)-1H-pyrazol-4-carboxamide (278) (180 mg, 0.57 mmol ), 2-(chloromethyl)-1H-1,3benzodiazol (189 mg, 1.13 mmol) and DIPEA (180 mg, 1.4 mmol) in DMF (8 mL) gave the title compound (66 mg, 25% ) after purification by basic prep-HPLC.

¹ H NMR (DMSO-J6, 500 MHz): δ [ppm] = 12.20 (s, 1H), 8.54 (t, J=5.7 Hz, 1H), 8.37 (dt , J=4.6, 1.2 Hz, 1H), 8.16 (s, 1H), 7.88 (s, 1H), 7.71-7.65 (m, 1H), 7.53 ( d, J=7.4 Hz, 1H), 7.45-7.36 (m, 2H), 7.18-7.07 (m, 2H), 5.75 (s, OH), 4.60 -4.54 (m, 2H), 4.12-3.99 (m, 2H), 3.67 (kv, J=14.0 Hz, 2H), 2.71-2.62 (m, 3H ), 2.17-2.04 (m, 2H), 1.96 (t, J=10.4 Hz, 1H), 1.67-1.58 (m, 1H), 1.54-1, 38 (m, 2H), 1.05-0.91 (m, 1H).

HPLC-MS (method c): [m/z]: 448.3 [M+H] ⁺ .

Ethyl 1-({ 1-[(tert-butoxy)carbonyl]azetidin-3-yl}methyl)-1 H-pyrazole-4-carboxylate (279)

- 194 039916

Ethyl 1H-pyrazole-4-carboxylate (0.55 g, 3.92 mmol), tert-butyl 3-(bromomethyl)azetidine-1-carboxylate (0.982 g, 3.92 mmol) and potassium carbonate (1.08 g, 8 mmol) in DMF (10 ml) was stirred at room temperature for 16 hours. The resulting mixture was diluted with water and extracted with ethyl acetate (3x50 ml). The combined organic extracts were washed with saturated sodium chloride solution (4x50 ml), dried (Na ₂ SO ₄ ), filtered and evaporated in vacuo to give the title compound (1.33 g) as a pale yellow oil.

1H NMR (CDCl3, 500 MHz): δ [ppm] = 7.92 (s, 1H), 7.90 (s, 1H), 4.38-4.26 (m, 4H), 4, 06 (t, J=9.0 Hz, 2H), 3.74 (dd, J=9.0, 5.1 Hz, 2H), 3.17-3.01 (m, 1H), 1.46 (s, 9H), 1.37 (t, 7= 7.1 Hz, 3H).

HPLC-MS (Method A): [m/z]: 310.15 [M+H]+.

1-({1-[(tert-Butoxy)carbonyl]azetidin-3-yl}methyl)-1H-pyrazole-4-carboxylic acid (280)

Similarly, using General Procedure 5, LiOH (2.06 g, 86 mmol) and ethyl 1-({1-[(tert-butoxy)carbonyl]azetidin-3-yl}methyl)-1H-pyrazole-4-carboxylate (279 ) (1.33 g, 4.3 mmol) in THF (30 mL) and water (30 mL) gave the title compound (1.02 g, 83%) as a white solid.

1H NMR (CDCl3, 500 MHz): δ [ppm] = 12.31 (s, 1H), 8.31 (s, 1H), 7.81 (s, 1H), 4.36 (d, J=7.3 Hz, 2H), 3.94-3.81 (m, 2H), 3.72-3.61 (m, 2H), 3.07-2.89 (m, 1H), 1 .37 (s, 9H).

HPLC-MS (method A): [m/z]: 304.05 [M+Na]+.

tert-Butyl 3-[(4-{[(3-fluoropyridin-2-yl)methyl]carbamoyl}-1H-pyrazol-1-yl)methyl]azetidine-1carboxylate(281)

Similarly, using the general procedure 6, 1-({1-[(tert-butoxy)carbonyl]azetidin-3yl}methyl)-1H-pyrazole-4-carboxylic acid (280) (500 mg, 1.78 mmol), (3-fluoropyridin-2-yl)methanamine dihydrochloride (A2) (389 mg, 1.96 mmol), DIPEA (1.1 ml, 6 mmol) and HATU (740 mg, 2 mmol) in DCM (30 ml) gave the title compound (624 mg, 89%) as a colorless oil after purification by flash column chromatography eluting with a gradient of 50-100% EtOAc/heptane followed by 0-10% MeOH/EtOAc.

1H NMR (CDCl3, 500 MHz): δ [ppm] = 8.32 (dt, J=4.7, 1.1 Hz, 1H), 7.84 (s, 1H), 7.79 ( s, 1H), 7.35 (m, 1H), 7.24-7.20 (m, 1H), 4.71 (dd, J=4.5, 1.3 Hz, 2H), 4.27 (d, J=7.6 Hz, 2H), 3.97 (t, J=8.6 Hz, 2H), 3.66 (dt, J=7.6, 3.9 Hz, 2H), 3 .02-2.98 (m, 1H), 1.36 (s, 9H).

HPLC-MS (method A): [m/z]: 390.1 [M+H] ⁺ .

1-(Azetidin-3-ylmethyl)-N-[(3-fluoropyridin-2-yl)methyl]-1H-pyrazole-4-carboxamide (282)

Similarly, using General Procedure 2, tert-butyl 3-[(4-{[(3-fluoropyridin-2yl)methyl]carbamoyl}-1H-pyrazol-1-yl)methyl]azetidine-1-carboxylate (281) ( 624 mg, 1.6 mmol) and 12 M HC (2 ml) in MeOH (20 ml) gave the free base of the title compound (248 mg, 54%) as a white solid after purification on an SCX-2 cartridge, with washing with DCM and MeOH, then eluting with 7N. a solution of ammonia in MeOH.

¹ H NMR (CDCla, 500 MHz): δ [ppm] = 8.42 (d, J=4.6 Hz, 1H), 7.94 (s, 1H), 7.88 (s, 1H ), 7.44 (t, J=9.3 Hz, 1H), 7.34-7.25 (m, 1H), 4.81 (d, J=4.2 Hz, 2H), 4.42 (d, J=7.4 Hz, 2H), 3.82 (t, J=8.2 Hz, 2H), 3.55-3.47 (m, 2H), 3.28 (m, 1H) .

HPLC-MS (method A): [m/z]: 290 [M+H] ⁺ .

1-{[1-(1H-1,3-Benzodiazol-2-ylmethyl)azetidin-3-yl]methyl }-N-[(3-fluoropyridin-2-yl)methyl]-1Hpyrazol-4-carboxamide (illustrative example no. 187)

Similarly, using General Procedure 3, 1-(azetidin-3-ylmethyl)-N-[(3-fluoropyridin-2yl)methyl]-1H-pyrazole-4-carboxamide (281) (50 mg, 0.17 mmol) , 1H-benzimidazole-2-carbaldehyde (37.9

- 195 039916 mg, 0.26 mmol), DIPEA (0.06 ml, 0.34 mmol) and MgSO ₄ (300 mg) in MeOH (5 ml) at room temperature for 18 h, followed by the addition of NaBH ₄ ( 7 mg, 0.17 mmol) gave the title compound (32 mg, 43%) as a light yellow solid after purification by basic prep-HPLC.

1H NMR (DMSO-d ₆ , 500 MHz): δ [ppm] = 12.08 (s, 1H), 8.54 (t, J=5.7 Hz, 1H), 8.37 (dt , J=4.6, 1.4 Hz, 1H), 8.18 (s, 1H), 7.87 (s, 1H), 7.68 (ddd, J=10.1, 8.3, 1 .3 Hz, 1H), 7.47 (s, 2H), 7.40 (dt, J=8.6, 4.4 Hz, 1H), 7.15-7.10 (m, 2H), 4 .57 (dd, J=5.6, 1.5 Hz, 2H), 4.35 (d, J=7.5 Hz, 2H), 3.77 (s, 2H), 3.38 (t, J=7.5 Hz, 2H), 3.12-3.05 (m, 2H), 2.89-2.79 (m, 1H).

HPLC-MS (method C): [m/z]: 420.1 [M+H]+.

1-({1-[2-(1H-1,3-Benzodiazol-2-yl)ethyl]azetidin-3-yl}methyl)-N-[(3-fluoropyridin-2-yl)methyl]-1Hpyrazol- 4-carboxamide [illustrative example No. 192] °

Cn ⁿ L ' ^N ^ ⁴ Ν

HN^I

M

Similarly, using General Procedure 8, 1-(azetidin-3-ylmethyl)-N-[(3-fluoropyridin-2yl)methyl]-1H-pyrazole-4-carboxamide (281) (200 mg, 0.69 mmol) , N(2-nitrophenyl)prop-2-enamide (D) (139.5 mg, 0.73 mmol) and DBU (0.12 ml, 0.83 mmol) in MeCN (10 ml) gave the crude intermediate, which reacted with iron powder (130 mg) in AcOH (3 ml). Purification by basic prep-HPLC followed by flash column chromatography (eluting with a 0-20% MeOH/DCM gradient) gave the title compound (67 mg, 25%) as a white solid.

1H NMR (Methanol-04, 500 MHz): δ [ppm] = 8.36 (dt, J=4.5, 1.3 Hz, 1H), 8.12 (s, 1H), 7, 93 (s, 1H), 7.61 (ddd, J=9.8, 8.5, 1.3 Hz, 1H), 7.52 (td, J=6.1, 3.1 Hz, 2H) , 7.40 (dt, J=8.5, 4.5 Hz, 1H), 7.24-7.17 (m, 2H), 4.73 (d, J=1.6 Hz, 2H), 4.37 (d, J=7.2 Hz, 2H), 3.45 (t, J=7.9 Hz, 2H), 3.14-3.07 (m, 2H), 3.00 (d q, J=13.8, 7.0 Hz, 1H), 2.94 (s, 4H).

HPLC-MS (method C): [m/z]: 434.2 [M+H] ⁺ .

The above general scheme 16:

methyl 1H-pyrazole-4-carboxylate (282) o

[NH

1H-pyrazole-4-carboxylic acid (20 g, 178.4 mmol) and sulfuric acid (39.65 ml) in MeOH (200 ml) were heated at 70°C for 20 h. The reaction mixture was cooled to room temperature and evaporated in a vacuum. An aqueous solution of NaOH was added, adjusting the pH to ~6. The aqueous layer was extracted with ethyl acetate (3x200 ml) and the combined organic extracts were dried (MgSO ₄ ), filtered and evaporated in vacuo to give the title compound (18.5 g, 78%) as a white solid.

1 H NMR (CDCl ₃ , 500 MHz): δ [ppm] = 8.10 (s, 2H), 3.87 (s, 3H).

HPLC-MS (method A): [m/z]: 126.85 [M+H] ⁺ .

Methyl 1-[1 -(diphenylmethyl)azetidin-3-yl]-1H-pyrazole-4-carboxylate (283)

A suspension of methyl 1H-pyrazole-4-carboxylate (282) (3.34 g, 26.48 mmol), 1(diphenylmethyl)azetidin-3-yl methanesulfonate (10.93 g, 34.43 mmol) and potassium carbonate (10 .98 g, 79 mmol) in DMF (70 ml) was heated at 100° C. for 3 hours. The reaction mixture was cooled to room temperature and diluted with water (150 ml). The resulting mixture was extracted with ethyl acetate (3x150 ml), the combined organic extracts were washed with saturated sodium chloride solution (5x100 ml), dried (Na ₂ SO ₄ ), filtered and evaporated in vacuum. The resulting residue was purified by flash chromatography through a pad of silica gel (~15 g) (eluting with 10-20% EtOAc/heptane), the eluent was evaporated in vacuo and the title compound (4.03 g, 44%) was obtained as a white solid after precipitation from a mixture of EtOAc/heptane.

1H NMR (CDCl3, 250 MHz): δ [ppm] = 8.14 (s, 1H), 7.96 (s, 1H), 7.49-7.42 (m, 4H), 7, 36-7.17 (m, 6H), 4.98 (m, 1H), 4.54 (s, 1H), 3.86 (s, 3H), 3.75-3.69 (td, J= 7.2, 1.5 Hz, 2H), 3.54-3.45 (m, 2H).

HPLC-MS (method A): [m/z]: 348.10 [M+H] ⁺ .

1-[1-(Diphenylmethyl)azetidin-3-yl]-1H-pyrazole-4-carboxylic acid (284)

- 196 039916

Similar to general procedure 5, methyl 1-[1-(diphenylmethyl)azetidin-3-yl]-1H-pyrazole-4carboxylate (283) (2.1 g, 6.04 mmol) and LiOH (1.45 g, 60 mmol ) in THF/water (20 ml/20 ml) gave the title compound (1.95 g, 93%) as a white solid.

1H NMR (CDCl ₃ , 250 MHz): δ [ppm] = 8.15 (s, 1H), 8.04 (s, 1H), 7.82-7.70 (m, 4H), 7 .51-7.37 (m, 6H), 5.65-5.55 (m, 1H), 5.55-5.42 (m, 1H), 4.84-4.69 (m, 2H) , 4.39-4.25 (m, 2H).

HPLC-MS (method A): [m/z]: 334.1 [M+H] ⁺ .

1-[1-(Diphenylmethyl)azetidin-3-yl]-X-[(3-fluoropyridin-2-yl)methyl]-1H-pyrazole-4-carboxamide (285)

Similar to general procedure 6, 1-[1-(diphenylmethyl)azetidin-3-yl]-1H-pyrazole-4-carboxylic acid (284) (1.18 g, 2.46 mmol), (3-fluoropyridine-2- yl)methanamine dihydrochloride (A2) (0.587 g, 2.95 mmol), DIPEA (1.71 ml, 9.82 mmol) and HATU (1.21 g, 3.19 mmol) in THF (50 ml) and DMF (10 mL) gave the title compound (1.06 g, 82%) as a yellow solid after purification by flash column chromatography (eluting with a 60-100% EtOAc/heptane gradient).

1H NMR (DMSO-06, 500 MHz): δ [ppm] = 8.59 (t, J=5.7 Hz, 1H), 8.40 (s, 1H), 8.37 (d, J=4.6Hz, 1H), 7.93(s, 1H), 7.68(t, J=9.3Hz, 1H), 7.47(m, 4H), 7.39(m, 1H), 7.29 (t, J=7.6 Hz, 4H), 7.19 (t, J=7.3 Hz, 2H), 5.04 (n, J=6.8 Hz, 1H) , 4.62-4.56 (m, 3H), 3.59 (t, J=7.7 Hz, 2H), 3.35 (s, 2H).

HPLC-MS (method A): [m/z]: 442.15 [M+H] ⁺ .

N-{5Н,6Н,7Н-cyclopenta[b]pyrudin-7-yl}-1-[1 -(diphenylmethyl)azetidin-3-yl]-1Н-pyrazole-4carboxamide (286)

Similar to general procedure 6 1-[1-(diphenylmethyl)azetidin-3-yl]-1H-pyrazole-4-carboxylic acid (284) (500 mg, 1.5 mmol), 5H,6H,7H-cyclopenta[b] pyridine-7-amine dihydrochloride (404 mg, 1.95 mmol), DIPEA (0.86 ml, 5 mmol) and HATU (0.74 g, 2 mmol) in DCM (20 ml) gave the crude title compound ( 949 mg) as a white residue, which was used in the next step without further purification.

HPLC-MS (method A): [m/z]: 450.15 [M+H]+.

1-(Azetidin-3-yl)-X-[(3-fluoropyridin-2-yl)methyl]-1H-pyrazole-4-carboxamide (287)

1-[1-(diphenylmethyl)azetidin-3-yl]-X-[(3-fluoropyridin-2-yl)methyl]-1H-pyrazole-4-carboxamide (285) (1.06 g, 2.01 mmol ) and Pd/C (10%) (0.20 g, 0.201 mmol) were suspended in EtOH (11 ml). A solution of TEA (0.849 mL, 6.04 mmol) and formic acid (0.228 mL, 6.04 mmol) in EtOH (11 mL) was added and the reaction mixture was stirred at reflux for 2 h. celite, and the resulting residue was washed with MeOH (10 ml). The filtrate was evaporated in vacuo to give the crude product. Purification on an SCX-2 cartridge, washing with DCM and MeOH, and eluting with 7N. Ammonia/MeOH solution gave the title compound (0.354 g, 56%) as a colorless oil.

1H NMR (DMSO-d ₆ , 500 MHz): δ [ppm] = 8.57 (t, J=5.6 Hz, 1H), 8.37 (d, J=4.6 Hz, 1H ), 8.34 (s, 1H), 7.93 (s, 1H), 7.72-7.65 (m, 1H), 7.40 (dd, J=8.5, 4.3 Hz, 1H), 5.18 (n, J=7.3 Hz, 1H), 4.58 (dd, J=5.6, 1.4 Hz, 2H), 3.84 (t, J=7.7 Hz, 2H), 3.73 (t, J=8.1 Hz, 2H).

HPLC-MS (method A): [m/z]: 276.1 [M+H] ⁺ .

1-(Azetidin-3-yl)-N-{5Н,6Н,7Н-cyclopenta[b]pyridin-7-yl}-1Н-pyrazol-4-carboxamide (288)

Crude N-{5H,6H,7H-cyclonent[b]nuridin-7-yl}-1-[1-(diphenylmethyl)azetidin-3-yl]-1H-pyrazole

- 197 039916

4-carboxamide (286) (949 mg) and palladium (10 wt% on carbon, 0.12 g, 0.11 mmol) were suspended in EtOH (20 ml). TEA (454 μl, 3 mmol), formic acid (125 μl, 3 mmol) in EtOH (20 ml) was added and the resulting mixture was heated at reflux for 1 hour. The reaction mixture was cooled to room temperature and filtered through a pad of celite. The resulting residue was washed with MeOH and the combined filtrates were evaporated in vacuo. The resulting residue was dissolved in a minimum amount of MeOH and purified by passing through an SCX-2 cartridge, washing with DCM and MeOH and eluting with 7N. a solution of ammonia in MeOH. Evaporation of the eluent gave the title compound (342 mg, quant) as a white solid.

¹ H NMR (DMSO-d ₆ , 500 MHz): δ [ppm] = 8.40-8.34 (m, 3H), 7.94 (s, 1H), 7.68 (d, J =7.6Hz, 1H), 7.22(dd, J=7.6, 4.9Hz, 1H), 5.48-5.40(m, 1H), 5.18(m, 1H) , 3.88-3.81 (m, 2H), 3.78-3.71 (m, 2H), 2.98 (ddd, J=16.0, 8.9, 3.0 Hz, 1H) , 2.86 (dt, J=16.0, 8.3 Hz, 2H), 2.47 (dd, J=6.5, 4.8 Hz, 1H), 1.95-1.84 (m , 1H).

HPLC-MS (method A): [m/z]: 284.05 [M+H]+.

1-[1-(1H-1,3-Benzodiazol-2-ylmethyl)azetidin-3-yl]-N-[(3-fluoropyridin-2-yl)methyl]-1H-pyrazole-4carboxamide [Illustrative Example No. 136]

Similarly, using General Procedure 3, 1-(azetidin-3-yl)-N-[(3-fluoropyridin-2yl)methyl]-1H-pyrazole-4-carboxamide (287) (196 mg, 0.71 mmol) , 1H-1,3-benzodiazol-2-carbaldehyde (125 mg, 0.85 mmol) and MgSO ₄ (100 mg) in MeOH (8 ml) at room temperature for 3 days, followed by the addition of NaBH ₄ (81 mg , 2.1 mmol) gave the title compound (82 mg, 28%) as a white solid after purification by basic prep-HPLC.

1H NMR (DMSO-d ₆ , 500 MHz): δ [ppm] = 12.32 (s, 1H), 8.58 (t, J=5.7 Hz, 1H), 8.43 (s , 1H), 8.39-8.37 (m, 1H), 7.96 (s, 1H), 7.69 (ddd, J=10.0, 8.3, 1.2 Hz, 1H), 7.61-7.45(m, 2H), 7.40(dt, J=8.6, 4.4Hz, 1H), 7.177.10(m, 2H), 5.06(m, 1H) , 4.59 (dd, J=5.7, 1.5 Hz, 2H), 3.92 (s, 2H), 3.85-3.81 (m, 2H), 3.61-3.57 (m, 2H).

HPLC-MS (Method C): [m/z]: 406.2 [M+H] ⁺ .

1-{1-[2-(1H-1,3-Benzodiazol-2-yl)ethyl]azetidin-3-yl}-N-[(3-fluoropyridin-2-yl)methyl]-1Hpyrazol-4-carboxamide (illustrative example #167)

Similar to general procedure 8, 1-(azetidin-3-yl)-N-[(3-fluoropyridin-2-yl)methyl]-1H-pyrazole-4carboxamide (287) (364 mg, 1.32 mmol), N- (2-nitrophenyl)prop-2-enamide (D) (254 mg, 1.32 mmol) and DBU (0.197 ml, 1.32 mmol) in MeCN (12 ml) gave a crude intermediate which was further reacted with AcOH (8 ml) and iron powder (62 mg) gave the title compound (104 mg, 43%) as a white solid after purification by basic prep-HPLC.

1H NMR (DMSO-d ₆ , 500 MHz): δ [ppm] = 12.15 (s, 1H), 8.56 (t, J=5.7 Hz, 1H), 8.37 (d , J=4.6 Hz, 1H), 8.34 (s, 1H), 7.93 (s, 1H), 7.71-7.65 (m, 1H), 7.49 (brs, 1H), 7.45-7.37 (m, 2H), 7.14-7.07 (m, 2H), 4.98 (p, J=6.8 Hz, 1H), 4.59-4 .56 (m, 2H), 3.69 (t, J=7.6 Hz, 2H), 3.38 (t, J=7.3 Hz, 2H), 2.94 (t, J=7, 1 Hz, 2H), 2.85 (t, J=7.0 Hz, 2H).

HPLC-MS (Method D): [m/z]: 420.3 [M+H] ⁺ .

1-{1-[2-(1H-1,3-Benzodiαzol-2-yl)ethyl]azetidin-3-yl}-N-{5H,6H,7H-cyclopenta[b]pyridin-7-yl}1 N-pyrazole-4-carboxamide (illustrative example No. 191)

Similar to general procedure 8, 1-(azetidin-3-yl)-N-{5H,6H,7H-cyclopenta[b]pyridin-7-yl}-1Hpyrazol-4-carboxamide (288) (342 mg, 1.21 mmol), N-(2-nitrophenyl)prop-2-enamide (D) (244 mg, 1.27 mmol) and DBU (0.22 ml, 1.5 mmol) in MeCN (10 ml) gave the crude intermediate , which reacted with iron powder (170 mg, 3 mmol) in AcOH (4 mL) to give the title compound (145 mg, 44%) as a white solid after purification by basic prep-HPLC followed by flash column chromatography (elution with a 0-20% MeOH/DCM gradient).

1H NMR (Methanol-d ₄ , 500 MHz): δ [ppm] = 8.37 (d, J=5.0 Hz, 1H), 8.26 (s, 1H), 8.01 (s , 1H), 7.78-7.73 (m, 1H), 7.52 (dd, J=5.4, 3.0 Hz, 2H), 7.30 (dd, J=7.6, 5 .0 Hz, 1H), 7.24-7.18 (m, 2H), 5.56 (t, J=8.2 Hz, 1H), 5.06 (m, 1H), 3.86 (td , J=7.3, 1.6 Hz, 2H), 3.64-3.58 (m, 2H), 3.11-3.07 (m, 3H), 3.03-2.92 (m , 3H), 2.69 (m, 1H),

- 198 039916

2.02 (m, 1H).

HPLC-MS (method C): [m/z]: 428.2 [M+H]+.

The above general scheme 18:

tert-butyl 2-(cyanomethyl)-1Н-1,3-benzodiazol-1-carboxylate (296)

2-(1H-1,3-benzodiazol-2-yl)acetonitrile (1 g, 6.36 mmol) and TEA (887 μl, 6.36 mmol) were dissolved in THF (20 ml) and BOC anhydride (1. 64 g, 7.51 mmol). The resulting mixture was stirred at room temperature for 16 h.

The reaction mixture was evaporated in vacuo. The resulting residue was dissolved in chloroform and washed with 1M aqueous HC solution (50 ml), saturated aqueous NaHCO ₃ solution (50 ml) and saturated sodium chloride solution (50 ml). The organic phase was dried (Na ₂ SO ₄ ), filtered and evaporated in vacuo. Purification by flash column chromatography eluting with a 10-30% EtOAc/heptane gradient gave the title compound (923 mg, 56%) as an orange solid after trituration in DCM/heptane.

1H NMR (CDCl3, 250 MHz): δ [ppm] = 8.00-7.91 (m, 1H), 7.84-7.74 (m, 1H), 7.48-7.36 (m, 2H), 4.37 (s, 2H), 1.77 (s, 9H).

HPLC-MS (Method A): [m/z]: 158.35 [M+H-Boc] ⁺ .

tert-Butyl 2-(2-{[(tert-butoxy)carbonyl]amino}ethyl)- 1H-1,3-benzodiazol-1-carboxylate (297)

Suspension of tert-butyl 2-(cyanomethyl)-1H-1,3-benzodiazol-1-carboxylate (296) (597 mg, 2.32 mmol), TEA (258 mg, 2.55 mmol), Boc ₂ O (557 mg, 2.55 mmol) and Pd (10 wt% on charcoal, 247 mg, 0.23 mmol) in EtOH (20 ml) was stirred under hydrogen atmosphere for 16 h. The reaction mixture was filtered through a pad of celite and the resulting residue was washed with MeOH and 7n. a solution of ammonia in MeOH. The combined filtrates were evaporated in vacuo. Purification by flash column chromatography eluting with a 0-60% EtOAc/heptane gradient gave the title compound (748 mg, 89%) as a white solid.

1H NMR (CDCl3, 250 MHz): δ [ppm] = 7.99-7.90 (m, 1H), 7.75-7.67 (m, 1H), 7.39-7.31 (m, 2H), 5.50 (s, 1H), 3.76 (q, J=5.7 Hz, 2H), 3.36 (t, J=5.7 Hz, 2H), 1.73 (s, 9H), 1.43 (s, 9H).

HPLC-MS (method A): [m/z]: 362.1 [M+H] ⁺ .

Ethyl 3-(bromomethyl)-1-methyl-1H-pyrazole-5-carboxylate (298) o / ^N ~N Br

Ethyl 1,3-dimethyl-1H-nirazol-5-carboxylate (433 mg, 2.57 mmol), NBS (0.623 g, 3.5 mmol) and AIBN (40 mg) in DCE (10 ml) was heated at 80° C for 1.5 hours. The reaction mixture was cooled to room temperature and evaporated in vacuo. Purification by flash column chromatography eluting with a 0-10% Et ₂ O/heptane gradient gave the title compound (419mg, 66%) as a white waxy solid.

1H NMR (CDCl3, 250 MHz): δ [ppm] = 6.90 (s, 1H), 4.48 (s, 2H), 4.36 (q, J=7.1 Hz, 3H) , 4.18 (s, 3H), 1.40 (t, J=7.1 Hz, 3H).

HPLC-MS (method A): [m/z]: 246.75 /248.75 [M+H] ⁺ .

tert-Butyl 2-(2-{[(tert-butoxy)carbonyl] ({[5-(ethoxycarbonyl)-1-methyl-1 H-pyrazol-3yl]methyl} )amino}ethyl)- 1H-1,3 -benzodiazol-1 -carboxylate (299)

tert-Butyl 2-(2-{[(tert-butoxy)carbonyl]amino}ethyl)-1H-1,3-benzodiazol-1-carboxylate (298) (263 mg, 0.73 mmol) in THF (5 ml ) were cooled in an ice bath. NaBH ₄ (60% in mineral oil, 60 mg, 1.5 mmol) was added and the resulting mixture was stirred for 5 minutes. Added ethyl 3

- 199 039916 (bromomethyl)-1-methyl-1H-pyrazole-5-carboxylate (297) (90 mg, 0.36 mmol) in THF (2 ml), the mixture was allowed to warm to room temperature and stirred for 10 min. The reaction was quenched with water dropwise, then further diluted with water and extracted with DCM (3x20 ml). The combined organic extracts were dried (Na ₂ SO 4 ), filtered and evaporated in vacuo. Purification by flash column chromatography eluting with a 0-80% EtOAc/heptane gradient gave the title compound (126 mg, 66%) as a colorless oil.

1H NMR (CDCl ₃ , 500 MHz): δ [ppm] = 7.68-7.62 (m, 1H), 7.32-7.27 (m, 1H), 7.18-7, 13 (m, 2H), 6.43 (s, 1H), 5.26 (s, 2H), 4.19 (q, J=7.1 Hz, 2H), 4.09-4.06 (m , 2H), 4.05 (s, 3H), 3.18 (dd, J=8.4, 6.8 Hz, 2H), 1.39 (s, 18H), 1.23 (t, J= 7.1 Hz, 3H).

HPLC-MS (method A): [m/z]: 528.2 [M+H]+.

3-({[(tert-Butoxy)carbonyl] (2-{1-[(tert-butoxy)carbonyl]-1H-1,3-benzodiazol-2 yl}ethyl)amino}methyl)-1-methyl-1 N-pyrazole-5-carboxylic acid (3 00)

Similar to general procedure 5, tert-butyl 2-(2-{[(tert-butoxy)carbonyl]({[5-(ethoxycarbonyl)-1methyl-1H-pyrazol-3-yl]methyl})amino}ethyl)-1H -1,3-benzodiazol-1-carboxylate (299) (126 mg, 0.24 mmol) and LiOH (17 mg, 0.72 mmol) in THF (5 ml) and water (5 ml) gave the title compound (152 mg) as a pale yellow oil.

HPLC-MS (method A): [m/z]: 500.15 [M+H] ⁺ .

tert-Butyl 2-(2-{[(tert-butoxy)carbonyl][(5-{[(3-fluoropyridin-2-yl)methyl]carbamoyl}-1-methyl1 H-pyrazol-3-yl)methyl] amino} ethyl)- 1H-1,3-benzodiazol-1-carboxylate (301)

Similar to general procedure 6, tert-butyl 2-(2-{[(tert-butoxy)carbonyl]({[5-(ethoxycarbonyl)-1methyl-1H-pyrazol-3-yl]methyl})amino}ethyl)-1H -1,3-benzodiazol-1-carboxylate (300) (130 mg, 0.26 mmol), (3-fluoropyridin-2-yl)methanamine dihydrochloride (A2) (57 mg, 0.29 mmol), DIPEA (150 μl, 0.86 mmol) and HATU (119 mg, 0.31 mmol) in DCM (10 ml) gave the crude title compound (317 mg, 30% pure) as a white solid, which was used in the next step without cleaning.

HPLC-MS (method A): [m/z]: 608.2 [M+H] ⁺ .

3-({[2-(1H-1,3-Benzodiazol-2-yl)ethyl)]amino} methyl)-N-[(3-fluoropyridin-2-yl)methyl]-1-methyl-1Hpyrazol-5 -carboxamide (illustrative example No. 220)

Similar to general procedure 2, tert-butyl 2-(2-{[(tert-butoxy)carbonyl][(5-{[(3-fluoropyridin-2yl)methyl]carbamoyl}-1-methyl-1H-pyrazole-3 -yl)methyl]amino}ethyl)-1H-1,3-benzodiazol-1-carboxylate (301) (30%, 317 mg, 0.16 mmol) and 12 M HC (0.63 ml) in MeOH (5 ml) at 60° C. for 15 min gave the title compound (44 mg, 69%) as a white solid after purification by basic prep-HPLC.

1H NMR (DMSO-H6, 500 MHz): δ [ppm] = 8.93 (m, 1H), 8.34 (dt, J=4.6, 1.4 Hz, 1H), 7, 72-7.63 (m, 1H), 7.56 (m, 1H), 7.52 (dd, J=7.0, 1.4 Hz, 1H), 7.43-7.33 (m, 1H), 7.21-7.11 (m, 2H), 6.71 (s, 1H), 5.41 (s, 2H), 4.574.49 (m, 2H), 4.00 (s, 3H ), 3.05-2.96 (m, 4H).

HPLC-MS (method C): [m/z]: 408.1 [M+H] ⁺ .

The above general scheme 19:

methyl 2-bromo-1-methyl-1Н-imidazole-4-carboxylate (302)

- 200 039916 about

MeO^>r ^N A

Ι %Br \

To a stirred solution of methyl 1-methyl-1H-imidazole-4-carboxylate (5 g, 46.38 mmol) in THF (100 mL) was added NBS (8.25 g, 46.38 mmol). The reaction mixture was allowed to stir at room temperature for 72 h. The resulting mixture was evaporated and the resulting crude residue was dissolved in EtOAc (80 ml), washed with saturated aqueous Na ₂ S ₂ O ₃ (100 ml) and NaOH solution pH ~ 12 ( 50 ml). The NaOH solution was extracted with 1:4 IPA:CHC3 (5x10 ml), the organic layers were combined, dried over MgSO ₄ , filtered and evaporated to give the crude product. Purification by column chromatography with a gradient of 0-100% EtOAc/heptane gave a purified product, which was dissolved in pH~12 NaOH solution (50 ml) and extracted with dichloromethane. The organic layers were combined, dried over MgSO ₄ , filtered and evaporated to give the title compound (5.58 g, 71%) as a white solid.

1 H NMR (DMSO-d ₆ , 250 MHz): δ [ppm] = 8.08 (s, 1H), 3.74 (s, 3H), 3.63 (s, 3H).

HPLC-MS (method A): [m/z]: 218.8, 220.75 [M+H]+.

Methyl 2-formyl-1-methyl-1H-imidazole-4-carboxylate (303) o

^MeOD <%_ N O \

A 2M solution of iPrMgCl in THF (3.31 ml, 6.62 mmol). After 20 minutes, the reaction mixture was cooled to -78° C. and DMF (1.08 ml, 13.96 mmol) was added dropwise. The reaction mixture was allowed to warm to room temperature, after 40 min the reaction was quenched with saturated NaHCO ₃ solution (10 ml) to give a thick white emulsion. The resulting product was extracted with ethyl acetate (5x10 ml). The combined organic layers were dried over MgSO ₄ , filtered and evaporated to give the crude product. Purification by flash column chromatography on silica gel with a gradient of 0-70% EtOAc/heptane gave the title compound (128 mg, 66%) as a white solid.

1H NMR (DMSO-de, 250 MHz): δ [ppm] = 9.74 (d, J=0.7 Hz, 1H), 8.26 (s, 1H), 3.96 (s, 3H), 3.80 (s, 3H).

HPLC-MS (method A): [m/z]: 168.9 [M+H] ⁺ .

Methyl 2-(1-hydroxy-2-nitroethyl)-1-methyl-1 H-imidazole-4-carboxylate (304)

To a stirred solution of methyl 2-formyl-1-methyl-1H-imidazole-4-carboxylate (303) (523 mg, 1.47 mmol), MeOH (12 ml) and nitromethane (78.9 µl, 1.47 mmol) while 1M NaOH solution (12 ml) was added to the flame and the reaction mixture was allowed to warm to room temperature throughout the experiment. After 1 h, the solution was acidified with 2M HC to ~pH 4 and evaporated to remove methanol. The aqueous layer was extracted with 1:4 IPA/SHC ₃ (4x10 ml) and the combined organic layers were dried over MgSO ₄ , filtered and evaporated to give the crude product which was purified by silica gel flash column chromatography with a gradient of 0-80% EtOAc/ heptane to give the final product (165 mg, 36%) as an off-white oil.

1H NMR (DMSO-de, 500 MHz): δ [ppm] = 7.94 (s, 1H), 6.35 (d, J=7.7 Hz, 1H), 5.40 (m, 1H), 5.13 (dd, J=13.3, 4.0 Hz, 1H), 4.90 (dd, J=13.3, 9.4 Hz, 1H), 3.75 (s, 3H ), 3.73 (s, 3H).

HPLC-MS (method A): [m/z]: 229.9 [M+H] ⁺ .

Methyl 1-methyl-2-[(E)-2-nitroethenyl]-1H-imidazole-4-carboxylate (305) o

M eO 'U- \ ^N ° ² \

A stirred solution of methyl 2-(1-hydroxy-2-nitroethyl)-1-methyl-1H-imidazole-4-carboxylate (304) (197 mg, 0.64 mmol) and acetic anhydride (3.5 ml) was heated to 45 °C. After 4 h the reaction mixture was evaporated, obtaining a yellow residue, which was divided between saturated aqueous NaHCO ₃ (20 ml) and EtOAc (20 ml). The organic layer was separated and the aqueous layer was extracted with ethyl acetate (4x5 ml). The combined organic layers were dried over MgSO ₄ , filtered and evaporated to give the crude product, purified by silica gel flash column chromatography with a gradient of 20-100% EtOAc/heptane to give the title product (97 mg, 60%) as a yellow solid.

1H NMR (DMSO-d6, 500 MHz): δ [ppm] = 8.19 (s, 1H), 8.02-7.93 (m, 2H), 3.89 (s, 3H), 3.78 (s, 3H).

HPLC-MS (method A): [m/z]: 211.9 [M+H]+.

- 201 039916

Methyl 2-(2-{[(tert-butoxy)carbonyl]amino}ethyl)-1-methyl-1Н-imidazole-4-carboxylate (306)

Into a nitrogen-purged and stirred solution of methyl 1-methyl-2-[(E)-2-nitroethenyl]-1H-imidazole4-carboxylate (305) (110 mg, 0.44 mmol), di-tert-butyl dicarbonate (386, 5 mg, 1.77 mmol) and EtOH (5 ml) was added Pd/C (10%) (47.1 mg, 0.04 mmol). The reaction mixture was purged (x3) with nitrogen, then with hydrogen. After 48 h the reaction mixture was filtered under vacuum through a glass filter using methanol (20 ml) to wash the filter cake and the filtrate was evaporated to give the crude product. The resulting crude product was purified by flash column chromatography (eluting with a 0-8% MeOH/DCM gradient) to give the purified product (75 mg, 26%) as a yellow solid.

HPLC-MS (method A): [m/z]: 284 [M+H]+.

2-(2-{[(tert-Butoxy)carbonyl]amino}ethyl)-1-methyl-1H-imidazole-4-carboxylic acid (307)

Similar to general procedure 5, methyl 2-(2-{[(tert-butoxy)carbonyl]amino}ethyl)-1-methyl-1Himidazole-4-carboxylate (306) (0.5 g, 0.79 mmol) and LiOH (0.19 g, 7.94 mmol) in THF (20 ml) and water (10 ml) gave the crude product as a yellow solid (0.703 g, quant.).

HPLC-MS (Method A): [m/z]: 270.05 [M+H]+.

tert-Butyl yl)ethyl]carbamate (308)

N-[2-(4-{[(3-Fluoropyridin-2-yl)methyl]carbamoyl}-1-methyl-1 H-imidazol-2-

Similar to general procedure 6, 2-(2-{[(tert-butoxy)carbonyl]amino}ethyl)-1-methyl-1H-imidazole4-carboxylic acid (307) (0.703 g, 0.79 mmol), HATU (0.604 g, 1.59 mmol), (3-fluoropyridin-2yl)methanamine dihydrochloride (A2) (0.316 g, 1.58 mmol) and DIPEA (0.968 ml, 5.56 mmol) in THF (20 ml) and DMF (5 ml) at room temperature for 3 h gave the title compound (0.092 g, 30%) as a yellow solid after purification by flash column chromatography (eluting with a 0-9% MeOH/DCM gradient).

HPLC-MS (method A): [m/z]: 378.15 [M+H] ⁺ .

2-(2-Aminoethyl)-N-[(3-fluoropyridin-2-yl)methyl]-1-methyl-1H-imidazole-4-carboxamide (309) °

C ⁿ 1 \

Similar to general procedure 2, tert-butyl N-[2-(4-{[(3-fluoropyridin-2-yl)methyl]carbamoyl}-1 methyl-1H-imidazol-2-yl)ethyl]carbamate (308) ( 94 mg, 0.25 mmol) and 12M HC (0.415 ml, 4.98 mmol) in

MeOH (4 ml) at 40°C for 2 h gave the product (9 mg, 13%) as a brown solid after flash column chromatography with 7M NHs/MeOH (x3).

HPLC-MS (Method D): [m/z]: 278.05 [M+H] ⁺ .

2-{2-[(1H-1,3-Benzodiazol-2-ylmethyl)amino]ethyl}-N-[(3-fluoropyridin-2-yl)methyl]-1-methyl-1nimidazole-4-carboxamide (illustrative example no. 210)

Similar to general procedure 3, 2-(2-aminoethyl)-N-[(3-fluoropyridin-2-yl)methyl]-1-methyl-1Himidazole-4-carboxamide (309) (9 mg, 0.032 mmol), anhydrous MgSO ₄ (150 mg), 1H-benzimidazole-2carbaldehyde (6.2 mg, 0.042 mmol) and DIPEA (11.3 μl, 0.065 mmol) in MeOH (2 ml) for 72 h, after addition of NaBH ₄ (1.8 mg, 0.049 mmol) gave the title compound (4.1 mg, 30%) as a brown solid after purification by flash column chromatography with a gradient of 050% MeOH/DCM.

1H NMR (MeOD, 500 MHz): δ [ppm] = 8.18 (d, J=4.7 Hz, 1H), 7.49-7.44 (m, 1H), 7.44 ( s, 1H), 7.42-7.37 (m, 2H), 7.28-7.22 (m, 1H), 7.11-7.06 (m, 2H), 4.63 (s, 2H), 3.97 (s, 2H), 3.53 (s, 3H), 2.97 (t, J=6.6 Hz, 2H), 2.81 (t, J=6.6 Hz, 2H).

HPLC-MS (method A): [m/z]: 408.1 [M+H] ⁺ .

The above general scheme 20:

- 202 039916 ethyl 1-(cyanomethyl)-5-methyl-1H-imidazole-4-carboxylate (310)

A solution of ethyl 4-methyl-1H-imidazole-5-carboxylate (3 g , 19.46 mmol) in DMF (25 ml). After 10 minutes, bromoacetonitrile (1.63 ml, 23.35 mmol) was added dropwise to the resulting solution, giving a red mixture. The reaction mixture was allowed to warm to room temperature. After 16 h the reaction mixture was evaporated and the resulting residue was diluted by adding saturated aqueous NaHCO ₃ (40 ml). The solution was extracted with ethyl acetate (6x20 ml), the combined organic layers were washed with saturated sodium chloride solution (4x15 ml), dried over MgSO ₄ , filtered and evaporated to give the crude product. Purification by flash column chromatography with a 0-6% MeOH/DCM gradient gave the title compound (1.3 g, 35%) as an orange solid.

Ή NMR (DMSO-J6, 500 MHz): δ [ppm] = 7.76 (s, 1H), 5.38 (s, 2H), 4.22 (kv, J=7.1 Hz, 2H), 2.52 (s, 3H), 1.27 (t, J=7.1 Hz, 3H).

HPLC-MS (method A): [m/z]: 194 [M+H]+.

Ethyl 1-(2-{[(tert-butoxy)carbonyl]amino}ethyl)-4-methyl-1H-imidazole-4-carboxylate (311)

Into a nitrogen-purged, stirred solution of ethyl 1-(cyanomethyl)-5-methyl-1H-imidazole-4-carboxylate (310) (0.72 g, 3.73 mmol), EtOH (20 ml), TEA (0.675 ml, 4.85 mmol) and di-tert-butyl dicarbonate (1.06 g, 4.85 mmol) Pd/C (10%) (0.397 g, 0.37 mmol) was added at room temperature. The reaction mixture was purged with nitrogen (x3) and then with hydrogen. After 16 h the reaction mixture was filtered through a glass filter and the filter cake was washed with 7M NH ₃ /MeOH (40 ml). The filtrate was evaporated to give crude product which was further purified by flash column chromatography using a 0-10% MeOH/DCM gradient to give the title product (1.07 g, 84%) as an off-white solid.

1H NMR (DMSO-d ₆ , 500 MHz): δ [ppm] = 7.53 (s, 1H), 7.01-6.93 (m, 1H), 4.19 (q, J= 7.1 Hz, 2H), 3.97 (t, J=5.9 Hz, 2H), 3.19 (kv, J=6.0 Hz, 2H), 2.43 (s, 3H), 1 .37-1.32 (m, 9H), 1.28-1.23 (m, 3H).

HPLC-MS (method A): [m/z]: 298.1 [M+H] ⁺ .

Lithium 1-(2-{[(tert-butoxy)carbonyl]amino}ethyl)-5-methyl-1H-imidazole-4-carboxylate (312)

To a stirred solution of ethyl 1-(2-{[(tert-butoxy)carbonyl]amino}ethyl)-5-methyl-1Himidazole-4-carboxylate (311) (1.07 g, 3.12 mmol), THF (35 ml) and water (10 ml) was added LiOH (0.224 g, 9.36 mmol), the reaction mixture was heated to 50°C. After 24 hours the reaction mixture was evaporated to give the lithium salt of the product (0.670 g) as a yellow solid.

HPLC-MS (method A): [m/z]: 270.05 [M+H] ⁺ .

tert-Butyl yl)ethyl]carbamate (313)

N-[2-(4-{[(3-fluoropyridin-2-yl)methyl]carbamoyl}-5-methyl-1 H-imidazol-1 -

As in general procedure 6, to lithium 1-(2-{[(tert-butoxy)carbonyl]amino}ethyl)-5-methyl-1nimidazole-4-carboxylate (312) (0.670 g, 2.488 mmol), THF (30 ml ), DMF (10 mL) and DIPEA (1.733 mL, 9.952 mmol) HATU (1.419 g, 3.732 mmol) and (3-fluoropyridin-2yl)methanamine dihydrochloride (0.743 g, 3.732 mmol) were added at room temperature. After 2 h the reaction mixture was evaporated and the resulting residue was dissolved in saturated aqueous NaHCO ₃ (100 ml) which was extracted with ethyl acetate (6x20 ml). The combined organic layers were washed with saturated sodium chloride solution (3x20 ml), dried over MgSO ₄ , filtered and evaporated to give the crude product, which was further purified by silica gel column chromatography with a gradient of 0-10% MeOH in DCM, obtaining an orange oil, further purified using column chromatography on kp-NH silica gel with a gradient of 0-100% EtOAc in heptane, obtaining the final product (0.320 g, 31%) as a yellow oil.

¹ H NMR (DMSO-a ₆ , 500 MHz) 5 [ppm] = 8.39 (d, J=4.6 Hz, 1H), 8.22 (t, J=5.4 Hz, 1H), 7.70 (t, J=9.3 Hz, 1H), 7.54 (s, 1H), 7.45-7.37 (m, 1H), 6.98 (t, J=5 .6 Hz, 1H), 4.60 (d, J=4.4 Hz, 2H), 3.96 (t, J=6.0 Hz, 2H), 3.19 (q, J=5.9 Hz, 2H), 2.44 (s, 3H), 1.39-1.20 (m, 9H).

HPLC-MS (Method A): [m/z]: 378.05 [M+H] ⁺ .

- 203 039916

-(2-Aminoethyl)-N -[(3-fluoropyridin-2-yl)methyl] -5-methyl-1 H-imidazole-4-carboxamide dihydrochloride (314)

Similar to general procedure 2, a solution of tert-butyl N-[2-(4-{[(3-fluoropyridin-2yl)methyl]carbamoyl}-5-methyl-1H-imidazol-1-yl)ethyl]carbamate (313 ) (0.320 g, 0.776 mmol) in MeOH (10 ml) at room temperature was added conc. HC (1.293 ml, 15.52 mmol), the reaction mixture was heated to 50°C. After 2 hours the reaction mixture was evaporated to give the title product (0.294 g, quant) as a yellow solid.

¹ H NMR (Methanol-d ₄ , 500 MHz): δ [ppm] = 9.03 (s, 1H), 8.54 (s, 1H), 8.06 (s, 1H), 7, 75 (s, 1H), 4.89 (s, 2H), 4.58 (t, J=6.5 Hz, 2H), 3.48 (t, J=6.4 Hz, 2H), 2, 67 (s, 3H).

HPLC-MS (Method A): [m/z]: 277.95 [M+H]+.

-(2-{[2-(1H-1,3-Benzodiazol-2-yl)ethyl]amino}ethyl)-N-[(3-fluoropyridin-2-yl)methyl]-5-methyl-1nimidazol-4 -carboxamide (illustrative example No. 219)

Similar to general procedure 8, 1-(2-aminoethyl)-N-[(3-fluoropyridin-2-yl)methyl]-5-methyl-1Himidazole-4-carboxamide dihydrochloride (314) (0.294 g, 0.77 mmol) , DBU (0.347 ml, 2.33 mmol) and N-(2nitrophenyl)prop-2-enamide (D) (0.332 g, 1.73 mmol) in MeCN (20 ml), after silica gel column chromatography (0-3 % MeOH in DCM) and further reaction with AcOH (2 ml) and iron powder (0.041 g) gave the title compound (0.030 g, 30%) as an off-white solid, after purification by basic prep-HPLC, followed by flash column chromatography (kp-NH, 0-5% MeOH/DCM gradient elution).

1H NMR (DMSO-d ₆ , 500 MHz): δ [ppm] = 12.09 (brs, 1H), 8.39 (d, J=4.7 Hz, 1H), 8.22 (t, J=5.5 Hz, 1H), 7.75-7.66 (m, 1H), 7.61 (s, 1H), 7.53-7.35 (m, 3H), 7, 13-7.06 (m, 2H), 4.60 (d, J=4.3 Hz, 2H), 3.97 (t, J=6.3 Hz, 2H), 3.00-2.93 (m, 2H), 2.94-2.88 (m, 2H), 2.84 (t, J=6.3 Hz, 2H), 2.47 (s, 3H).

HPLC-MS (method d): [m/z]: 422.1 [M+H] ⁺ .

The above general scheme 21:

ethyl 2-{1-[(tert-butoxy)carbonyl]-3-chloroazetidin-3-yl}-5-chloro-1,3-thiazole-4-carboxylate (321)

Ethyl 2-{1-[(tert-butoxy)carbonyl]azetidin-3-yl}-1,3-thiazole-4-carboxylate (161) (1.15 g, 3.68 mmol) and hexachloroethane (0.83 g, 3 mmol) was dissolved in THF (40 ml) and cooled to -78°C. 2M NaHMDS (1.75 ml) was added dropwise and the reaction mixture was allowed to stir at -78°C for 30 min. More 2M NaHMDS (0.6 ml) was added and the reaction mixture was allowed to stir at -78°C for another 30 minutes.

The reaction was quenched with saturated aqueous NH ₄ Cl, warmed to room temperature, diluted with water and extracted with DCM (3x80 ml). The combined organic extracts were dried (Na ₂ SO ₄ ), filtered and evaporated in vacuo. Two main components have been identified in the raw product. Purification by flash column chromatography (elution with a 2-15% EtOAc/heptane gradient) gave the title compound (456 mg, 32%) as a pale yellow oil.

1 H NMR (CDCl ₃ , 500 MHz): δ [ppm] = 4.72 (d, J=9.2 Hz, 2H), 4.37 (q, J=7.1 Hz, 2H), 4.32 (m, 2H), 1.40 (s, 9H), 1.35 (t, J=7.1 Hz, 3H).

HPLC-MS (method A): [m/z]: 402.9 [M+Na]+.

2-{1-[(tert-Butoxy)carbonyl]-3- ^{chloroazetidin} -3-yl}-5-chloro-1,3-thiazole-4-carboxylic acid (322) NBoc 01^ ³

As in general procedure 5, ethyl 2-{1-[(tert-butoxy)carbonyl]-3-chloroazetidin-3-yl}-5-chloro1,3-thiazole-4-carboxylate (321) (450 mg, 1.18 mmol) and LiOH (0.08 g, 4 mmol) in THF (5 ml) and water (5 ml) gave the crude title compound (461 mg) as a colorless oil which crystallized on standing.

- 204 039916

1H NMR (CDC13, 250 MHz): δ [ppm] = 4.76 (dd, J=9.6, 1.0 Hz, 2H), 4.45 (dd, J=9.6, 1 .0 Hz, 2H), 1.50 (s,

9H).

HPLC-MS (method A): [m/z]: 378.85 [M+Na]+.

tert-Butyl 3-chloro-3-(5-chloro-4-{[(3-fluoropyridin-2-yl)methyl]carbamoyl}-1,3-thiazol-2-yl)azetidine1-carboxylate(323)

As in General Procedure 6, 2-{1-[(tert-butoxy)carbonyl]-3-chloroazetidin-3-yl}-5-chloro-1,3thiazole-4-carboxylic acid (322) (460 mg, 1.3 mmol), (3-fluoropyridin-2-yl)methanamine dihydrochloride (A2) (311 mg, 1.56 mmol), DIPEA (0.75 ml, 4 mmol) and HATU (0.59 g, 2 mmol) in DCM (10 ml) gave the title compound (479 mg, 80%) after purification by flash column chromatography (eluting with a 10-50% EtOAc/heptane gradient).

1H NMR (CDC13, 500 MHz): δ [ppm] = 8.34 (dt, J=4.7, 1.1 Hz, 1H), 8.26-8.20 (m, 1H), 7.38-7.31(m, 1H), 7.23-7.16(m, 1H), 4.75(m, 2H), 4.70(m, 2H), 4.35(m, 2H), 1.40 (s, 9H).

HPLC-MS (Method A): [m/z]: 460.95 [M+H]+.

5-Chloro-2-(3-chloroazetidin-3-yl)-N-[(3-fluoropyridin-2-yl)methyl]-1,3-thiazole-4-carboxamide (324)

Similar to general procedure 2, tert-butyl 3-chloro-3-(5-chloro-4-{[(3-fluoropyridin-2yl)methyl]carbamoyl}-1,3-thiazol-2-yl)azetidine-1-carboxylate (323) (479 mg, 1.04 mmol) and 12M HC (2 ml) in MeOH (20 ml) gave the title compound (375 mg, quant) as a white solid after purification on an SCX-2 cartridge, with washing with DCM and MeOH, then eluting with 7N. a solution of ammonia in MeOH.

1H NMR (DMSO-de, 250 MHz): δ [ppm] = 9.05-8.96 (m, 1H), 8.40 (dt, J=4.6, 1.4 Hz, 1H ), 7.78-7.66 (m, 1H), 7.48-7.37 (m, 1H), 4.66 (dd, J=5.8, 1.6 Hz, 2H), 4, 41 (d, J=10.5 Hz, 2H), 4.07 (d, J=10.5 Hz, 2H).

HPLC-MS (method A): [m/z]: 360.8 [M+H] ⁺ .

2-{1-[2-(1H-1,3-benzodiazol-2-yl)ethyl]azetidin-3-yl}-5-chloro-N-[(3-fluoropyridin-2-yl)methyl]-1 ,3thiazol-4-carboxamide (Illustrative Example No. 209) and 2-{1-[2-(1H-1,3-benzodiazol-2-yl)ethyl]-3hydroxyazetidin-3-yl}-5-chloro-N- [(3-fluoropyridin-2-yl)methyl]-1,3-thiazole-4-carboxamide (illustrative example No. 211)

As per general procedure 8,5-chloro-2-(3-chloroazetidin-3-yl)-N-[(3-fluoropyridin-2-yl)methyl]1,3-thiazole-4-carboxamide (324) (375 mg , 1.04 mmol), N-(2-nitrophenyl)prop-2-enamide (D) (219 mg, 1.14 mmol) and DBU (0.17 ml, 1.14 mmol) in MeCN (20 ml) gave a crude mixture which was reacted with iron powder (0.2 g) in AcOH (5 ml). Purification by basic prep-HPLC followed by flash column chromatography (elution with a 0-20% MeOH/DCM gradient) gave the title compound (Illustrative Example #209) (60 mg, 14%) as a white solid.

1H NMR (DMSO-de, 500 MHz): δ [ppm] = 12.14 (s, 1H), 8.77 (t, J=5.6 Hz, 1H), 8.38 (dt, J=4.6, 1.3 Hz, 1H), 7.75-7.67 (m, 1H), 7.56-7.37 (m, 3H), 7.12 (dd, J=5, 9, 2.6 Hz, 2H), 4.66-4.60 (m, 2H), 3.89 (ddd, J=13.1, 7.4, 5.6 Hz, 1H), 3.61 (t, J=7.2 Hz, 2H), 2.94-2.88 (m, 2H), 2.87-2.81 (m, 2H).

One CH2 signal overlapped with water signals.

HPLC-MS (method C): [m/z]: 471.1 [M+H] ⁺ .

The by-product was also isolated by basic prep-HPLC to give 2-{1-[2-(1H-1,3benzodiazol-2-yl)ethyl]-3-hydroxyazetidin-3-yl}-5-chloro-N-[( 3-Fluoropyridin-2-yl)methyl]-1,3-thiazole-4carboxamide (Illustrative Example No. 211) (2 mg) as a white solid.

1H NMR (DMSO-de, 500 MHz): δ [ppm] = 12.16 (s, 1H), 8.86 (t, J=5.5 Hz, 1H), 8.35 (dt, J= 4.6, 1.4 Hz, 1H), 7.74-7.67 (m, 1H), 7.54-7.48 (m, 1H), 7.44-7.34 (m, 2H), 7.15-7.07 (m, 2H), 4.65 (d, J=5.5 Hz, 2H), 3.70 (d, J=8.3 Hz, 2H), 3, 43 (d, J=8.3 Hz, 2H), 3.01 (t, J=7.2 Hz, 2H), 2.87 (t, J=7.2 Hz, 2H).

HPLC-MS (Method C): [m/z]: 487.1 [M+H] ⁺ .

The above general scheme 22:

tert-butyl (1R,5S,6S)-6-carbamoyl-3-azabicyclo[3.1.0]hexane-3-carboxylate(325)

- 205 039916

Similar to general procedure 12, (1R,5S,6S)-3-[(tert-butoxy)carbonyl]-3-azabicyclo[3.1.0]hexane6-carboxylic acid (1 g, 4.4 mmol), TEA ( 1.04 ml, 0.01 mol), isobutyl chloroformate (0.86 ml, 0.01 mol) and 28% aqueous NH3 (1.33 ml, 0.07 mol) in THF (15 ml) gave crude the title product (1.20 g) as a pale yellow solid. This substance was directly used in the next step.

HPLC-MS (method A): [m/z]: 249.05 [M+Na]+.

tert-Butyl (1R,5S,6S)-6-carbamothioyl-3-azabicyclo[3.1.0]hexane-3-carboxylate (326)

Similar to general procedure 11, tert-butyl (1R,5S,6S)-6-carbamoyl-3-azabicyclo[3.1.0]hexane-3carboxylate (325) (1.2 g, 5.3 mmol) and Lawsson's reagent (1.18 g, 2.9 mmol) in DCM (40 mL) gave the title compound (939 mg, 73%) as a white foam after purification by flash column chromatography eluting with a 0-10% EtOAc/heptane gradient.

HPLC-MS (method A): [m/z]: 242.95 [M+H] ⁺ .

tert-Butyl carboxylate (327) (1R,5S,6R)-6-[4-(ethoxycarbonyl)-1,3-thiazol-2-yl]-3-azabicyclo[3.1.0]hexane-3-

Similar to general procedure 1, tert-butyl (1R,5S,6S)-6-carbamothioyl-3-azabicyclo[3.1.0]hexane3-carboxylate (326) (939 mg, 3.87 mmol), ethyl 3-bromo -2-oxopropanoate (0.53 ml, 4 mmol) and CaCO ₃ (0.21 g, 2 mmol) in EtOH (20 ml) gave the title compound (546 mg, 42%) as a colorless residue after purification by the method flash column chromatography eluting isocratically with 30% EtOAc/heptane followed by a second flash column chromatography eluting with a 040% EtOAc/heptane gradient.

HPLC-MS (method A): [m/z]: 339.1 [M+H] ⁺ .

2-[(1R,5S,6R)-3-[(tert-Butoxy)carbonyl]-3-azabicyclo[3.1.0]hexan-6-yl]-1,3-thiazole-4-carboxylic acid( 328)

Similar to general procedure 5, tert-butyl (1R,5S,6R)-6-[4-(ethoxycarbonyl)-l,3-thiazol-2-yl]-3azabicyclo[3.1.0]hexane-3-carboxylate ( 327) (546 mg, 1.61 mmol) and LiOH (0.12 g, 5 mmol) in THF (20 mL) and water (10 mL) gave the crude title compound (500 mg, 1.61 mmol) in as a pale yellow oil which was used in the next step without purification.

HPLC-MS (method A): [m/z]: 311.1 [M+H] ⁺ .

tert-Butyl (1 R,5S,6S)-6-(4-{[(3-fluoropyridin-2-yl)methyl]carbamoyl}-1,3-thiazol-2-yl)-3azabicyclo[3,1, 0]hexane-3-carboxylate (329)

F

H

Similar to the general procedure 6, 2-[(1R,5S,6R)-3-[(tert-butoxy)carbonyl]-3azabicyclo[3.1.0]hexan-6-yl]-1,3-thiazol-4- carboxylic acid (328) (500 mg, 1.61 mmol), (3fluoropyridin-2-yl)methanamine dihydrochloride (A2) (352.7 mg, 1.77 mmol), DIPEA (0.93 ml, 5.31 mmol ) and HATU (0.74 g, 1.93 mmol) in DCM (30 mL) gave the title compound (650 mg, 96%) as a colorless foam after purification by flash column chromatography eluting with a gradient of 20-100 % EtOAc/heptane.

HPLC-MS (method A): [m/z]: 419.1 [M+H] ⁺ .

2-[(1R,5S,6S)-3-azabicyclo[3.1.0]hexan-6-yl]-N-[(3-fluoropyridin-2-yl)methyl]-1,3-thiazole-4carboxamide (330)

F

H

Similar to general procedure 4, tert-butyl (1R,5S,6S)-6-(4-{[(3-fluoropyridin-2yl)methyl]carbamoyl}-1,3-thiazol-2-yl)-3-azabicyclo[ 3.1.0]hexane-3-carboxylate (329) (624 mg, 1.49

- 206 039916 mmol) and 12M HC (2 ml) in MeOH (20 ml) gave the title compound (475 mg, 83%) after purification on an SCX-2 cartridge, washing with DCM and MeOH, then eluting with 7N. a solution of ammonia in MeOH.

1H NMR (DMSO-a ₆ , 500 MHz): δ [ppm] = 8.65 (t, J=5.6 Hz, 1H), 8.39 (dt, J=4.6, 1, 3 Hz, 1H), 8.03 (s, 1H), 7.75-7.69 (m, 1H), 7.45-7.39 (m, 1H), 4.67-4.63 (m , 2H), 3.05 (m, 3H), 2.78 (d, J=11.4 Hz, 2H), 2.38 (t, J=3.3 Hz, 1H), 2.03-2 .00 (m, 2H).

HPLC-MS (method A): [m/z]: 319.05 [M+H] ⁺ .

2-[( 1 R,5S,6S)-3 -(1H-1,3 -Benzodiazol-2-ylmethyl)-3 -azabicyclo[3.1.0]hexan-6-yl] -N-[(3 fluoropyridin-2-yl)methyl]-1,3-thiazole-4-carboxamide (331)

Similar to the general procedure 3, 2-[(1R,5S,6S)-3-azabicyclo[3.1.0]hexan-6-yl]-N-[(3-fluoropyridin2-yl)methyl]-1,3- thiazole-4-carboxamide (330) (200 mg, 0.63 mmol), N-1,3-benzodiazol-2-carbaldehyde (110.17 mg, 0.75 mmol), DIPEA (0.33 ml, 2 mmol ) and MgSO ₄ (300 mg) in MeOH (10 ml) at room temperature for 18 h, followed by addition of NaBH ₄ (48 mg, 1.3 mmol) gave the title compound (62 mg, 22%) in as a pale yellow solid after trituration of the resulting residue in 1:1 DMSO/MeCN, followed by washing with MeOH.

1H NMR (DMSO-d6, 500 MHz): δ [ppm] = 12.17 (s, 1H), 8.63 (t, J=5.7 Hz, 1H), 8.40 (d, J=4.6Hz, 1H), 8.04(s, 1H), 7.76-7.67(m, 1H), 7.55(d, J=7.8Hz, 1H), 7, 46 (d, J=7.8 Hz, 1H), 7.44-7.39 (m, 1H), 7.20-7.09 (m, 2H), 4.66 (d, J=4, 9 Hz, 2H), 3.91 (s, 2H), 3.15 (d, J=9.1 Hz, 2H), 2.96 (t, J=2.8 Hz, 1H), 2.66 -2.60 (m, 2H), 2.14-2.10 (m, 2H).

HPLC-MS (Method C): [m/z]: 449.1 [M+H] ⁺ .

2-[( 1 R,5S,6S)-3 -(1H-1,3 -Benzodiazol-2-ylmethyl)-3 -azabicyclo[3.1.0]hexan-6-yl] -N-[(3 fluoropyridin-2-yl)methyl]-1,3-thiazole-4-carboxamide trihydrochloride (illustrative example No. 213)

2-[( 1 R,5S,6S)-3 -(1H-1,3-benzodiazol-2-ylmethyl)-3-azabicyclohexan-6-yl]-N-[(3[3.1.0] fluoropyridin-2-yl)methyl]-1,3-thiazole-4-carboxamide (331) (20 mg, 0.44 mmol) was suspended in MeOH (3 ml) and 12M HC (0.5 ml) was added, resulting in to dissolution. The reaction mixture was stirred for 1 h, evaporated in vacuo to give the title compound (25 mg, quant) as a yellow solid.

1H NMR (MeOD, 500 MHz): δ [ppm] = 8.58-8.55 (m, 1H), 8.19-8.13 (m, 1H), 8.05 (s, 1H ), 7.86-7.78 (m, 3H), 7.66-7.61 (m, 2H), 4.90 (s, 2H), 4.52 (s, 2H), 3.60- 3.52 (m, 2H), 3.22-3.13 (m, 2H), 3.11 (t, J=3.1 Hz, 1H), 2.43 (s, 2H).

HPLC-MS (Method C): [m/z]: 449.1 [M+H] ⁺ .

2-[(1R,5S,6S)-3-[2-(1H-1,3-Benzodiazol-2-yl)ethyl]-3-azabicyclo[3.1.0]hexan-6-yl]-N -[(3fluoropyridin-2-yl)methyl]-1,3-thiazole-4-carboxamide (illustrative example No. 218)

Similar to the general procedure 8, 2-[(1R,5S,6S)-3-azabicyclo[3.1.0]hexan-6-yl]-N-[(3-fluoropyridin2-yl)methyl]-1,3- thiazole-4-carboxamide (330) (264 mg, 0.83 mmol), N-(2-nitrophenyl)prop-2-enamide (D) (175 mg, 0.91 mmol) and DBU (0.14 ml, 0.91 mmol) in MeCN (15 mL) gave a crude intermediate which was reacted with iron powder (180 mg, 3.2 mmol) in AcOH (5 mL) to give the title compound (4 mg, 1%) in as a colorless residue after successive purification by flash chromatography (elution gradient 0-40% MeOH/DCM), basic prep-HPLC and flash column chromatography (elution gradient 0-5% MeOH/DCM).

Ή NMR (MeOD, 500 MHz): δ [ppm] = 8.38 (d, J=4.7 Hz, 1H), 7.94 (s, 1H), 7.61 (ddd, J= 9.7, 8.4, 1.2 Hz, 1H), 7.53 (m, 2H), 7.43-7.38 (m, 1H), 7.22 (m, 2H), 4.77 (d, J=1.5 Hz, 2H), 3.30 (m, 2H), 3.09 (m, 2H), 3.01 (m, 2H), 2.66 (m, 1H), 2 .58 (m, 2H), 2.15 (s, 2H).

HPLC-MS (method C): [m/z]: 463.1 [M+H] ⁺ .

- 207 039916

General scheme 23

2-(2-{[2-(1H-1,3-Benzodiazol-2-yl)ethyl](methyl)amino}ethyl)-N-[(3-fluoropyridin-2-yl)methyl]-1,3oxazole -4-carboxamide (illustrative example No. 223)

2-(2-{[2-(1H-1,3-benzodiazol-2-yl)ethyl]amino}ethyl)-N-[(3-fluoropyridin-2-yl)methyl]-1,3-oxazol- 4-carboxamide (illustrative example #127) (59 mg, 0.14 mmol), TEA (60 µl, 0.43 mmol) in DMF (1 ml) was stirred at room temperature for 1 h. MeI (8.9 µl, 0.14 mmol) and stirred at room temperature for 24 hours. The reaction mixture was again treated with MeI (62.9 μl, 0.79 mmol) and TEA (80 μl, 0.58 mmol) and allowed to stir at room temperature for 2 days. The resulting mixture was evaporated vigorously in vacuo to give a yellow oil (200 mg) which was purified by basic prep-HPLC followed by kp-NH column chromatography (0-10% MeOH/DCM gradient elution) and flash column chromatography (gradient elution 020% MeOH/DCM) to give the title compound (13 mg, 21%) as an off-white solid.

¹ H NMR (MeOD, 500 MHz): δ [ppm] = 8.34-8.29 (m, 1H), 8.15 (s, 1H), 7.61-7.54 (m, 1H), 7.44 (brs, 2H), 7.38-7.33 (m, 1H), 7.19-7.13 (m, 2H), 4.69 (d, J=1, 6 Hz, 2H), 3.09-3.04 (m, 2H), 3.04-2.99 (m, 2H), 2.982.91 (m, 4H), 2.39 (s, 3H).

HPLC-MS (method d): [m/z]: 423.1 [M+H]+.

The above general scheme 25:

tert-butyl 3-[(3-hydroxy-1-methoxy-1-oxopropan-2-yl)carbamoyl]azetidine-1-carboxylate (3 3 9) no-^ o

OV ^N G/\ ^N \-NBoc OMe

A stirred solution of methyl serinate hydrochloride (1:1) (2.3 g, 14.78 mmol) and TEA (2.27 ml, 16.26 mmol) in DCM (150 ml) was cooled to 0°C. 1-[(tert-Butoxy)carbonyl]azetidine-3-carboxylic acid (2.97 g, 14.78 mmol) was added followed by portions of DCC (3.36 g, 16.26 mmol) after which the reaction mixture was allowed to warm to room temperature. temperature. After 24 h the reaction mixture was evaporated and dissolved in EtOAc (~250 ml). The reaction mixture was allowed to stir at 50° C. for 45 minutes. The precipitate was then filtered off and the filtrate was evaporated to give the crude product as a white solid. Purification by flash column chromatography eluting with a gradient of 100% TBME followed by DCM/MeOH gave the title compound (4.5 g, 91%) as a yellow solid.

1H NMR (CDCl ₃ , 500 MHz): δ [ppm] = 6.75 (d, 1H), 4.66 (dt, 1H), 4.14-4.02 (m, 4H), 3 .99 (dd, 1H), 3.88 (d, 1H), 3.77 (s, 3H), 3.29 (dd, 1H), 1.42 (s, 9H).

HPLC-MS (method A): [m/z]: 246.95 [M-tBu] ⁺ .

Methyl 2-{1-[(tert-butoxy)carbonyl]azetidin-3-yl}-4,5-dihydro-1,3-oxazole-4-carboxylate (340) o

MeO'Ny^N < ^X X~<fjNBoc o

Similar to general procedure 14, tert-butyl 3-[(3-hydroxy-1-methoxy-1-oxopropan-2yl)carbamoyl]azetidine-1-carboxylate (339) (4.5 g, 13.4 mmol) and DAST ( 2.3 ml, 17.42 mmol) in DCM (120 ml) gave the title compound (3.3 g, 78%) as a pale yellow oil. The compound was used in the next step without further purification.

1H NMR (CDCl3, 500 MHz): δ [ppm] = 4.78 (m, 1H), 4.60-4.53 (m, 1H), 4.47 (m, 1H), 4, 12 (m, 4H), 3.80 (s, 3H), 3.48-3.36 (m, 1H), 1.43 (s, 9H).

Methyl 2-{1-[(tert-butoxy)carbonyl]azetidin-3-yl}-1,3-oxazole-4-carboxylate (341)

- 208 039916

Similar to general procedure 15, methyl 2-{1-[(tert-butoxy)carbonyl]azetidin-3-yl}-4,5-dihydro1,3-oxazole-4-carboxylate (340) (3.3 g, 10 45 mmol), DBU (2.21 ml, 14.77 mmol) and bromo(trichloro)methane (2.57 ml, 26.12 mmol) in DCM (25 ml) gave the title compound (2.3 g, 70%) as a yellow oil after purification by flash column chromatography using a gradient of 0-10% MeOH in DCM.

1H NMR (CDCl ₃ , 500 MHz): δ [ppm] = 8.21 (s, 1H), 4.30-4.24 (m, 4H), 3.96-3.89 (m, 4H), 1.44 (s, 9H).

HPLC-MS (Method A): [m/z]: 305.00 [M+Na]+.

2-{1-[(tert-Butoxy)carbonyl]azetidin-3-yl}-1,3-oxazole-4-carboxylic acid (342)

Similar to general procedure 5, methyl 2-{1-[(tert-butoxy)carbonyl]azetidin-3-yl}-1,3-oxazol-4carboxylate (341) (2.3 g, 7.33 mmol) and LiOH ( 0.26 g, 0.01 mol) in THF (35 ml) and water (8.5 ml) at room temperature for 4 hours gave the title compound (2.0 g, 86%) as a pale yellow oil. The resulting crude product was used in the next step without further purification.

HPLC-MS (method A): [m/z]: 290.95 [M+Na]+.

2-{1-[(tert-Butoxy)carbonyl]azetidin-3-yl}-1,3-oxazole-4-carboxylic acid (343)

Similar to general procedure 6, tert-butyl 3-{4-[(pyridin-2-ylmethyl)carbamoyl]-1,3-oxazol-2yl}azetidine-1-carboxylate (342) (1 g, 3.17 mmol), pyridin-2-ylmethanamine (0.343 g, 3.17 mmol), TEA (0.42 ml, 3.17 mmol) and HATU (1.81 g, 4.75 mmol) in DMF (10 ml) at room temperature in gave the title compound (1.07 g, 32%) as a brown oil over 48 hours after purification by flash column chromatography (eluting with a 40-80% EtOAc/heptane gradient).

¹ H NMR (CDCl ₃ , 500 MHz): δ [ppm] = 8.55-8.51 (m, 1H), 8.11 (s, 1H), 7.84 (s, 1H), 7.61 (td, J=7.7, 1.8 Hz, 1H), 7.25 (d, J=7.8 Hz, 1H), 7.15 (dd, J=7.4, 4, 9 Hz, 1H), 4.67 (d, J=5.5Hz, 2H), 4.22 (t, J=8.8 Hz, 2H), 4.16 (dd, J=8.6, 6 .2 Hz, 2H), 3.86-3.78 (m, 1H), 1.39 (s, 9H).

HPLC-MS (method A): [m/z]: 359.10 [M+H] ⁺ .

tert-Butyl carboxylate (344)

3-(4-{[(3-Fluoropyridin-2-yl)methyl]carbamoyl}-1,3-oxazol-2-yl)azetidine-1 -

Similar to general procedure 6, 2-{1-[(tert-butoxy)carbonyl]azetidin-3-yl}-1,3-oxazole-4carboxylic acid (342) (1.0 g, 3.48 mmol), (3 -fluoropyridin-2-yl)methanamine dihydrochloride (A2) (0.83 g, 4.17 mmol), DIPEA (2.42 ml, 13.91 mmol) and HATU (1.59 g, 4.17 mmol) in THF (50 ml) and DMF (10 ml) gave the title compound (1.56 g, 90%) as a yellow oil after purification by flash column chromatography (eluting with a 20-100% EtOAc/heptane gradient).

1H NMR (DMSO-d ₆ , 250 MHz): δ [ppm] = 8.68-8.54 (m, 2H), 8.38 (d, J=4.6 Hz, 1H), 7 .75-7.63(m, 1H), 7.40(m, 1H), 4.66-4.57(m, 2H), 4.31-4.15(m, 2H), 4.11 -3.95 (m, 3H), 1.39 (s, 9H).

HPLC-MS (method A): [m/z]: 377.15 [M+H] ⁺ .

tert-Butyl 3-{4-[(pyridazin-3-ylmethyl)carbamoyl]-1,3-oxazol-2-yl}azetidine-1-carboxylate (345)

Similar to general procedure 6, 2-{1-[(tert-butoxy)carbonyl]azetidin-3-yl}-1,3-oxazole-4carboxylic acid (342) (0.8 g, 2.98 mmol), 1- (pyridazin-3-yl)methanamine (0.42 g, 3.28 mmol, 85% purity), DIPEA (779 μl, 4.47 mmol) and HATU (1.36 g, 3.58 mmol) in DCM ( 20 ml) gave the title compound (0.90 g, 84%) as a brown residue after purification by flash column chromatography (gradient elution 50-100% EtOAc/heptane followed by 0-20% MeOH/EtOAc).

HPLC-MS (method M): [m/z]: 360.00 [M+H] ⁺ .

2-(Azetidin-3-yl)-N-(pyridin-2-ylmethyl)-1,3-Oksazol-4-carboxamide (346)

Similar to general procedure 2, tert-butyl 3-{4-[(pyridin-2-ylmethyl)carbamoyl]-1,3-oxazol-2-209 039916 yl}azetidine-1-carboxylate (343) (1.07 g, 2.99 mmol) and 12M HC (2 ml) in MeOH (20 ml) at 50°C for 30 min gave the title compound (0.457 g, 59%) as a yellow solid after purification on an SCX-2 cartridge (10 g), washing with DCM and MeOH, and then eluting with 7N.

NH ₃ /MeOH. The main eluent was then evaporated to give the title compound as the free base.

HPLC-MS (method A): [m/z]: 259.00 [M+H]+.

2-(Azetidin-3-yl)-N-[(3-fluoropyridin-2-yl)methyl]-1,3-oxazol-4-carboxαmide dihydrochloride (347)

Similar to general procedure 2, tert-butyl 3-(4-{[(3-fluoropyridin-2-yl)methyl]carbamoyl}-1,3oxazol-2-yl)azetidine-1-carboxylate (344) (1.56 g , 3.12 mmol) and conc. HC (5.2 ml, 62.51 mmol) in MeOH (25 ml) at room temperature for 16 hours gave the title compound (1.31 g, quant) as an off-white solid. The resulting crude product was used in the next step without purification.

HPLC-MS (method A): [m/z]: 277.05 [M+H] ⁺ .

2-(Azetidin-3-yl)-N-(pyridazin-3-ylmethyl)-1,3-oxazol-4-carboxamide (348)

Similar to general procedure 2, tert-butyl 3-{4-[(pyridazin-3-ylmethyl)carbamoyl]-1,3-oxazol-2yl}azetidine-1-carboxylate (345) (900 mg, 2.5 mmol) and TFA (10 ml, 130.6 mmol) in DCM (10 ml) at room temperature over 40 min gave the title compound (447 mg, 69%) as a beige solid after purification on an SCX-2 cartridge (10 g ), washing with DCM and MeOH and then eluting with 7N. NH ₃ /MeOH. The main eluent was then evaporated to give the title compound as the free base.

1H NMR (DMSO-d ₆ , 250 MHz): δ [ppm] = 9.14 (dd, J=4.7, 1.7 Hz, 1H), 8.95 (t, J=6, 0 Hz, 1H), 8.60 (s, 1H), 7.67 (dd, J=8.5, 4.7 Hz, 1H), 7.59 (dd, J=8.5, 1.8 Hz, 1H), 4.73 (d, J=6.2 Hz, 2H), 4.13-3.99 (m, 1H), 3.893.70 (m, 4H).

HPLC-MS (method M): [m/z]: 260.00 [M+H] ⁺ .

2-{1-[2-(1H-1,3-benzodiazol-2-yl)ethyl]azetidin-3-yl}-N-(pyridin-2-ylmethyl)-1,3-oxazole-4carboxamide (illustrative example No. 207)

Similar to general procedure 8, 2-(azetidin-3-yl)-N-(pyridin-2-ylmethyl)-1,3-oxazol-4carboxamide (346) (457 mg, 1.769 mmol), N-(2-nitrophenyl) prop-2-enamide (D) (408 mg, 2.123 mmol) and DBU (0.32 ml, 2.123 mmol) in MeCN (20 ml) gave a crude intermediate which was purified by flash column chromatography (eluting with a 0- 5% MeOH/DCM). The resulting intermediate was further reacted with iron powder (134 mg) in AcOH (3 mL) to give the title compound (26 mg, 8%) as a white solid after purification by basic prep-HPLC.

1H NMR (DMSO-d ₆ , 500 MHz): δ [ppm] = 8.50 (d, J=4.3 Hz, 1H), 8.33 (s, 1H), 7.80 (m , 1H), 7.50 (s, 2H), 7.40 (d, J=7.9 Hz, 1H), 7.34-7.29 (m, 1H), 7.22-7.16 ( m, 2H), 4.67 (s, 2H), 3.88 (n, J=7.3 Hz, 1H), 3.75 (t, J=8.0 Hz, 2H), 3.54 ( t, J=7.5 Hz, 2H), 3.05-2.99 (m, 2H), 2.99-2.93 (m, 2H).

HPLC-MS (method d): [m/z]: 403.2 [M+H] ⁺ .

2-{1-[2-(1H-1,3-Benzodiazol-2-yl)ethyl]azetidin-3-yl}-N-[(3-fluoropyridin-2-yl)methyl]-1,3-oxazol4 -carboxamide (illustrative example No. 208)

Similar to general procedure 8, 2-(αzetidin-3-yl)-N-[(3-fluoropyridin-2-yl)methyl]-1,3-oxazole-4carboxamide dihydrochloride (347) (0.657 g, 1.571 mmol), DBU (0.703 ml, 4.713 mmol) and N-(2nitrophenyl)prop-2-enamide (D) (0.332 g, 1.728 mmol) in MeCN (30 ml) at room temperature over 4 h gave a crude intermediate which was purified by column flash chromatography (eluting with a 0-3% MeOH/DCM gradient) to give a yellow oil (0.778 g). It was further reacted with iron powder (0.191 g) in AcOH (4 mL) at 80°C for 2 hours to give the title compound (0.072 g, 15%) as a white solid after column purification. flash chromatography (kp-NH, 0-3% MeOH/DCM gradient elution), followed by flash column chromatography (0-20% MeOH/DCM gradient elution).

1H NMR (MeOD, 500 MHz): δ [ppm] = 8.36 (d, J=4.7 Hz, 1H), 8.32 (s, 1H), 7.62-7.56 ( m, 1H), 7.50 (brs, 2H), 7.41-7.36 (m, 1H), 7.22-7.17 (m, 2H), 4.75 (d, J= 1.5 Hz, 2H), 3.88 (n, J= 7.3 Hz, 1H), 3.75 (t, J=8.0 Hz, 2H), 3.53 (t, J=7, 5 Hz, 2H), 3.04-2.99 (m, 2H), 2.99-2.93 (m, 2H).

HPLC-MS (Method C): [m/z]: 421.1 [M+H]+.

2-{1-[2-(1H-1,3-Benzodiazol-2-yl)ethyl]azetidin-3-yl}-N-(pyridazin-3-ylmethyl)-1,3-oxazol-4carboxamide (illustrative example No. 244)

Similar to general procedure 8, 2-(azetidin-3-yl)-N-(pyridazin-3-ylmethyl)-1,3-oxazol-4carboxamide (348) (440 mg, 1.7 mmol), N-(2- nitrophenyl)prop-2-enamide (D) (326 mg, 1.7 mmol) and DBU (279 μl, 1.87 mmol) in MeCN (20 ml) gave a crude intermediate which was reacted with iron powder (302 mg, 5.41 mmol) in AcOH (5 mL) to give the title compound (82 mg, 15%) as a white solid after purification by basic prep-HPLC.

1H NMR (MeOD, 500 MHz): δ [ppm] = 9.12 (dd, J=4.5, 2.1 Hz, 1H), 8.36 (s, 1H), 7.76- 7.69 (m, 2H), 7.51 (s, 2H), 7.24-7.19 (m, 2H), 4.88 (s, 2H), 3.93-3.85 (m, 1H), 3.78-3.74 (m, 2H), 3.58-3.53 (m, 2H), 3.07-3.01 (m, 2H), 3.01-2.95 ( m, 2H).

HPLC-MS (method B): [m/z]: 404.2 [M+H] ⁺ .

2-{1-[2-(4-Fluoro-1H-1,3-benzodiazol-2-yl)ethyl]azetidin-3-yl}-N-[(3-fluoropyridin-2-yl)methyl]-1 ,3oxazole-4-carboxamide (illustrative example No. 239)

Similar to the general procedure 8, 2-(azetidin-3-yl)-N-[(3-fluoropyridin-2-yl)methyl]-1,3-oxazol-4carboxamide; bis(trifluoroacetic acid) (347) (700 mg, 1.39 mmol), DBU (1.04 ml, 6.94 mmol) and N(3-fluoro-2-nitrophenyl) prop-2-enamide (G) ( 292 mg, 1.39 mmol) in MeCN (30 mL) gave a crude intermediate which was reacted with iron powder (173 mg, 3.1 mmol) in AcOH (5 mL) to give the title compound (98 mg, 29 %) as a pale pink solid after purification by flash column chromatography (eluting with a gradient of 0-8% MeOH/DCM).

1H NMR (MeOD, 500 MHz): δ [ppm] = 8.38 (dt, J=4.6, 1.1 Hz, 1H), 8.35 (s, 1H), 7.62 ( ddd, J=9.8, 8.4, 1.2 Hz, 1H), 7.41 (dt, J=8.6, 4.5 Hz, 1H), 7.31 (d, J=8, 1 Hz, 1H), 7.18 (td, J=8.1, 4.8 Hz, 1H), 6.95 (dd, J=10.8, 8.0 Hz, 1H), 4.77 ( d, J=1.5 Hz, 2H), 3.94-3.86 (m, 1H), 3.78 (t, J=7.9 Hz, 2H), 3.57 (t, J=7 .5 Hz, 2H), 3.08-3.03 (m, 2H), 3.02-2.97 (m, 2H).

HPLC-MS (method B): [m/z]: 439.2 [M+H] ⁺ .

2-{1-[2-(4-Fluoro-1H-1,3-benzodiazol-2-yl)ethyl]azetidin-3-yl}-N-(pyridin-2-ylmethyl)-1,3-oxazol4- carboxamide (illustrative example No. 258)

Similar to general procedure 8, 2-(azetidin-3-yl)-N-(pyridin-2-ylmethyl)-1,3-oxazol-4-carboxamide (346) (0.88 g, 3.21 mmol), N-( 3-fluoro-2-nitrophenyl)prop-2-enamide (G) (0.80 g, 3.53 mmol) and DBU (580 μl, 3.86 mmol) in MeCN (30 ml) gave a crude intermediate which reacted with iron powder (0.57 g, 10.24 mmol) in AcOH (16 ml) to give the title compound (0.59 g, 55%) as an off-white solid after purification by flash column chromatography (elution in a 0-15% MeOH/DCM gradient) followed by a basic prep HPLC.

1H NMR (MeOD, 500 MHz): δ [ppm] = 8.51-8.48 (m, 1H), 8.33 (s, 1H), 7.80 (td, J=7.7 , 1.8 Hz, 1H), 7.40 (d, J=7.9 Hz, 1H), 7.34-7.27 (m, 2H), 7.16 (td, J=8.1, 4.8 Hz, 1H), 6.92 (dd, J=10.8, 8.1 Hz, 1H), 4.67 (s, 2H), 3.93-3.83 (m, 1H), 3.75 (t, J=7.9 Hz, 2H), 3.54 (t, J=7.5 Hz, 2H), 3.05-3.00 (m, 2H), 2.99-2 .95 (m, 2H).

HPLC-MS (method C): [m/z]: 421.3 [M+H] ⁺ .

2-{1-[2-(4-Fluoro-1H-1,3-benzodiazol-2-yl)ethyl]azetidin-3-yl}-N-(pyridazin-3-ylmethyl)-1,3oxazol-4- carboxamide (illustrative example No. 261)

- 211 039916

Similar to general procedure 8, 2-(azetidin-3-yl)-N-(pyridazin-3-ylmethyl)-1,3-oxazol-4carboxamide (348) (417 mg, 1.61 mmol), N-(3- fluoro-2-nitrophenyl)prop-2-enamide (G) (372 mg, 1.77 mmol) and DBU (264 μl, 1.77 mmol) in MeCN (50 ml) gave a crude intermediate which was reacted with iron powder (261 mg, 4.67 mmol) in AcOH (10 mL) to give the title compound (268 mg,

54%) as a white solid after purification by basic prep-HPLC.

1H NMR (MeOD, 500 MHz): δ [ppm] = 9.12 (dd, J=4.5, 2.1 Hz, 1H), 8.36 (s, 1H), 7.76- 7.70 (m, 2H), 7.31 (d, J=8.1 Hz, 1H), 7.18 (td, J=8.1, 4.8 Hz, 1H), 6.97-6 .91 (m, 1H), 4.88 (s, 2H), 3.89 (n, J=7.3 Hz, 1H), 3.793.74 (m, 2H), 3.58-3.53 ( m, 2H), 3.06-3.02 (m, 2H), 3.02-2.96 (m, 2H).

HPLC-MS (method B): [m/z]: 422.2 [M+H]+.

N-[(3-Fluoropyridin-2-yl)methyl]-2-(1-{2-[7-(trifluoromethyl)-1H-1,3-benzodiazol-2yl]ethyl}azetidin-3-yl)-1 ,3-oxazole-4-carboxamide (illustrative example No. 273)

Similar to general procedure 8, 2-(azetidin-3-yl)-N-[(3-fluoropyridin-2-yl)methyl]-1,3-oxazole-4-carboxamide (347) (393 mg, 1.42 mmol), N-[2-nitro-6-(trifluoromethyl)phenyl]prop-2-enamide (K8) (474 mg, 1.42 mmol, 78% pure) and DBU (0.23 ml, 1.56 mmol) in MeCN (18 mL) gave a crude intermediate which was reacted with iron powder (280 mg, 5.01 mmol) in AcOH (8 mL) to give the title compound (197 mg, 32%) as a light brown foam after purification by flash column chromatography (elution with a gradient of 0-20% MeOH/DCM).

¹ H NMR (DMSO-66, 500 MHz): δ [ppm] = 12.64 (s, 1H), 8.55 (s, 1H), 8.52 (t, J=5.7 Hz , 1H), 8.39-8.35(m, 1H), 7.76(s, 1H), 7.71-7.65(m, 1H), 7.48-7.43(m, 1H ), 7.43-7.37 (m, 1H), 7.30-7.26 (m, 1H), 4.66-4.58 (m, 2H), 3.81 (n, J=7 .4 Hz, 1H), 3.63 (t, J=7.5 Hz, 2H), 3.35 (t, J=7.2 Hz, 2H), 2.90 (s, 4H).

HPLC-MS (Method D): [m/z]: 489.1 [M+H]+.

The above general scheme 26:

2-(3-chloropropyl)-1-{[2-(trimethylsilyl)ethoxy]methyl}-1Н-1,3-benzodiazol (349)

[2-(chloromethoxy)ethyl](trimethyl)silane (553 µl, 3.12 mmol) was added to a solution of 2-(3chloropropyl)-1H-1,3-benzodiazol hydrochloride (555 mg, 2.4 mmol) and DIPEA ( 962 μl, 5.52 mmol) in THF (25 ml). The reaction mixture was stirred at room temperature for 18 h, then extinguished with saturated aqueous NaHCO ₃ and was extracted with ethyl acetate (3x80 ml). The combined organic extracts were washed with saturated sodium chloride solution (50 ml), dried (Na ₂ SO ₄ ) and evaporated in vacuo. Purification by flash column chromatography (eluting with a 050% EtOAc/heptane gradient) gave the title compound (571 mg, 73%) as a pale yellow oil.

1H NMR (CDCl ₃ , 250 MHz): δ [ppm] = 7.85-7.75 (m, 1H), 7.53-7.44 (m, 1H), 7.38-7, 31 (m, 2H), 5.57 (s, 2H), 3.78 (t, J=6.1 Hz, 2H), 3.60 (dd, J=8.6, 7.7 Hz, 2H ), 3.21 (t, J=7.3 Hz, 2H), 2.59-2.44 (m, 2H), 0.96 (dd, J=8.6, 7.7 Hz, 2H) , 0.00 (s, 9H).

HPLC-MS (method M): [m/z]: 325.50 [M+H] ⁺ .

Methyl 2-(azetidin-3-yl)-1,3-oxazole-4-carboxylate hydrochloride (350)

Similar to general procedure 4, methyl 2-{1-[(tert-butoxy)carbonyl]azetidin-3-yl}-1,3-oxazole-4carboxylate (341) (634 mg, 2.25 mmol) and 12M HC (0 .89 ml) in MeOH (20 ml) at 60° C. gave the title compound (201 mg, 41%) as a white solid after trituration in DCM/Et ₂ O.

1H NMR (DMSO-H6, 250 MHz): δ [ppm] = 8.91 (s, 1H), 4.36-4.15 (m, 5H), 3.83 (s, 3H).

HPLC-MS (method M): [m/z]: 183.20 [M+H] ⁺ .

Methyl 2-{1-[3-(1-{[2-(trimethylsilyl)ethoxy]methyl}-1H-1,3-benzodiazol-2-yl)propyl]azetidin-3yl}-1,3-oxazol-4 -carboxylate (351)

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Suspension of methyl 2-(azetidin-3-yl)-1,3-oxazol-4-carboxylate hydrochloride (350) (300 mg, 1.37 mmol), 2-(3-chloropropyl)-1-{[2-( trimethylsilyl)ethoxy]methyl}-1H-1,3-benzodiazole (349) (758 mg, 2.33 mmol), DIPEA (837 µl, 4.8 mmol) and KI (228 mg, 1.37 mmol) in DMF (10 ml) was stirred at room temperature for 7 days. The reaction was quenched with saturated aqueous NaHCO ₃ and extracted with ethyl acetate (3x50 ml). The combined organic extracts were washed with saturated sodium chloride solution (4x50 ml), dried (Na ₂ SO ₄ ), filtered and evaporated in vacuo. Purification by flash column chromatography (gradient elution 40-100%

EtOAc/heptane then 2-40% MeOH/EtOAc) gave the title compound (329 mg, 46%, 91% pure) as a yellow residue.

HPLC-MS (method M): [m/z]: 471.15 [M+H]+.

2-{1-[3-(1-{[2-(Trimethylsilyl)ethoxy]methyl}-1H-1,3-benzodiazol-2-yl)propyl]azetidin-3-yl}-1,3oxazol-4- carboxylic acid (352)

Similar to general procedure 5, 2-{1-[3-(1-{[2-(trimethylsilyl)ethoxy]methyl}-1H-1,3-benzodiazol2-yl)propyl]azetidin-3-yl}-1,3 -oxazole-4-carboxylate (351) (329 mg, 0.7 mmol) and LiOH (50 mg, 2.1 mmol) in THF (10 mL) and water (10 mL) gave the title compound (363 mg, 91%, 80% pure) as a yellow residue. The compound was used in the next step without purification.

HPLC-MS (method M): [m/z]: 457.10 [M+H]+.

N-[(3-fluoropyridin-2-yl)methyl]-2-{1-[3-(1-{[2-(trimethylsilyl)ethoxy]methyl}- 1H-1,3-benzodiazol2-yl)propyl] azetidin-3 -yl} -1,3-oxazole-4-carboxamide (353)

Similar to general procedure 6, 2-{1-[3-(1-{[2-(trimethylsilyl)ethoxy]methyl}-1H-1,3-benzodiazol2-yl)propyl]azetidin-3-yl}-1,3 -oxazole-4-carboxylic acid (352) (363 mg, 0.64 mmol, 80% purity), (3-fluoropyridin-2-yl)methanamine dihydrochloride (A2) (152 mg, 0.76 mmol), DIPEA ( 388 µl, 2.23 mmol) and HATU (290 mg, 0.76 mmol) in DCM (20 ml) gave the title compound (302 mg, 84%) as a pale yellow residue after purification by flash column chromatography (elution with a 0-40% MeOH/DCM gradient).

HPLC-MS (method M): [m/z]: 565.15 [M+H]+.

2-{1-[3-(1H-1,3-Benzodiazol-2-yl)propyl]azetidin-3-yl}-N-[(3-fluoropyridin-2-yl)methyl]-1,3oxazol-4 -carboxamide (illustrative example No. 233)

TFA (4 ml, 52.23 mmol) was added to a solution of N-[(3-fluoropyridin-2-yl)methyl]-2-{1-[3-(1-{[2(trimethylsilyl)ethoxy]methyl}- 1H-1,3-benzodiazol-2-yl)propyl]azetidin-3-yl}-1,3-oxazole-4carboxamide (353) (300 mg, 0.53 mmol) in DCM (4 ml). The reaction mixture was stirred at room temperature for 3 hours. The reaction mixture was evaporated in vacuo, the resulting residue was dissolved in water, making basic by adding 2M aqueous NaOH solution. The resulting mixture was then extracted with 4:1 CHC ₃ /IPA (4x50 ml). The combined organic extracts were dried (Na ₂ SO ₄ ), filtered and evaporated in vacuo. The resulting residue was dissolved in a minimum volume of MeOH and loaded onto an SCX-2 cartridge (10 g). The cartridge was washed with DCM followed by MeOH and eluted with 7N. NH ₃ /MeOH.

The main eluent was evaporated in vacuo. Purification of the resulting residue by basic prep-HPLC gave the title compound (75 mg, 33%) as a white solid.

¹ H NMR (MeOD, 500 MHz): δ [ppm] = 8.37 (dt, J=4.6, 1.1 Hz, 1H), 8.33 (s, 1H), 7.61 (ddd, J=9.8, 8.4, 1.2 Hz, 1H), 7.50 (s, 2H), 7.41 (dt, J=8.7, 4.4 Hz, 1H), 7.23-7.17 (m, 2H), 4.77 (d, J=1.5 Hz, 2H), 3.93-3.84 (m, 1H), 3.75 (t, J= 8.0 Hz, 2H), 3.53 (t, J=7.5 Hz, 2H), 2.94 (t, J=7.6 Hz, 2H), 2.66-2.60 (m, 2H), 1.97-1.88 (m, 2H).

HPLC-MS (method B): [m/z]: 435.1 [M+H]+.

The above general scheme 28:

3-{[(tert-butoxy)carbonyl]amino}-3-methylbutanoic acid (360)

ΙΗΟ,^-χ,ΝΗΒοο o'

1M KOH (15.19 mL) was added to a mixture of 3-amino-3-methylbutanoic acid (1.78 g, 15.19 mmol) and Boc2O (3.48 g, 15.95 mmol) in 1,4-dioxane ( 30 ml) and stirred at room temperature for 40 h. The solvent was evaporated and diluted with water (60 ml). 1M LiOH was added to pH 13. The aqueous phase was extracted with Et ₂ O (3x30 ml) then the pH was adjusted to 3 with 2M HC and extracted with ethyl acetate (4x60 ml). The combined organic layers were washed with saturated sodium chloride solution (30 ml), dried (Na ₂ SO ₄ ), filtered and evaporated to give the title compound (2.13 g, 65%) as an off-white solid. The resulting compound was used in the next step without further purification.

¹ H NMR (CDCl ,, 250 MHz): δ [ppm] = 5.07 (s, 1H), 2.75 (s, 2H), 1.44 (s, 9H), 1.40 (s, 6H).

- {[(tert-Butoxy)carbonyl]amino} -2,2-dimethylpropanoic acid (361)

1M KOH (52.08 ml, 52.08 mmol) was added to a mixture of 3-amino-2,2-dimethylpropanoic acid hydrochloride (4 g, 26.04 mmol) and Boc ₂ O (5.97 g, 27.34 mmol ) in 1,4-dioxane (80 ml) and stirred at room temperature for 20 hours. The solvent was evaporated and diluted with water (50 ml). 1M LiOH was added to pH 13. The aqueous phase was extracted with Et ₂ O (3x50 ml) then the pH was adjusted to 3 with 2M HC and extracted with ethyl acetate (4x50 ml). The combined organic layers were washed with saturated sodium chloride solution (50 mL), dried (Na2SO4), filtered and evaporated to give the title compound (5.38 g, 95%) as a white solid. The resulting compound was used in the next step without further purification.

¹ H NMR (CDCI3, 250 MHz): δ [ppm] = 5.01 (s, 1H), 3.29-3.18 (m, 2H), 1.45 (d, 9H), 1 .23 (s, 6H).

tert-Butyl N-(1-carbamoyl-2-methylpropan-2-yl)carbamate (362)

IH ₂ N./-\.NHBoc o'

TEA (2.82 ml, 20.27 mmol) was added to an ice-cold (0°C) solution of 3-{[(tert-butoxy)carbonyl]amino}3-methylbutanoic acid (360) (2.59 g, 11.92 mmol) in THF (30 ml). The reaction mixture was stirred for 20 min, then 2-methylpropyl carbochloridate (2.78 ml, 17.88 mmol) was added dropwise at 0°C. The reaction mixture was stirred for 1 h, then a 35% aqueous solution of NH ₃ (3.09 ml, 66.76 mmol) was added dropwise, the reaction mixture was warmed to room temperature and stirred for 18 h. Saturated aqueous NaHCO solution was added ₃ (50 ml) and the aqueous layer was extracted with DCM (3x50 ml). The combined organic layers were dried (MgSO ₄ ), filtered and evaporated to give a pale yellow oil (4.11 g). Purification by flash column chromatography (eluting with a 0-10% MeOH-DCM gradient) gave the title compound (1.7 g, 55%, 84% pure) as a clear oil which solidified on standing.

¹ H NMR (CDCl ₃ , 250 MHz): δ [ppm] = 5.85 (br s, 1H), 5.39 (br s, 1H), 4.89 (br s, 1H ), 2.64 (s, 2H), 1.42 (s, 9H), 1.39 (s, 6H).

HPLC-MS (method A): [m/z]: 238.95 [M+Na]+.

tert-Butyl N-(2-carbamoyl-2,2-dimethylethyl)carbamate (363)

TEA (5.73 mL, 41.08 mmol) was added to an ice cold (0°C) solution of 3-{[(tert-butoxy)carbonyl]amino}2,2-dimethylpropanoic acid (361) (5.25 g, 24 .16 mmol) in THF (50 ml). The reaction mixture was stirred for 20 min, then 2-methylpropyl carbochloridate (4.7 ml, 36.3 mmol) was added dropwise at 0°C. The reaction mixture was stirred for 1 h, then a 35% solution of NH ₃ (7.48 ml, 135.31 mmol) was added dropwise. The reaction mixture was warmed to room temperature and stirred for 22 hours. Saturated aqueous NaHCO ₃ solution (100 ml) was added and the aqueous layer was extracted

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DXM (3x150 ml). The combined organic layers were dried (MgSO4), filtered and evaporated to give an off-white semi-solid (6.61 g). Purification by flash column chromatography (eluting with a 0-10% MeOH/DCM gradient) gave the title compound (3.7 g, 71%) as a white solid.

¹ H NMR (CDCl ₃ , 250 MHz): δ [ppm] = 5.95 (s, 1H), 5.30 (s, 1H), 5.08 (s, 1H), 3.25 ( e, J=6.6 Hz, 2H), 1.43 (s, 9H), 1.21 (s, 6H).

HPLC-MS (Method A): [m/z]: 239.10 [M+Na]+.

tert-Butyl N-(1-carbamothioyl-2-methylpropan-2-yl)carbamate (364)

IH ₂ Nv^\^NHBoc S \

Similar to general procedure 11, Lawsson's reagent (1.65 g, 4.08 mmol) and tert-butyl N-(1-carbamoyl-2-methylpropan-2-yl)carbamate (362) (1.7 g, 6.6 mmol, 84% pure) in DCM (50 ml) at room temperature for 19 hours gave the title compound (0.914 g, 60%) as a yellow oil after purification by flash column chromatography (eluting with a gradient of 0-50% EtOAc/ heptane) which solidified on standing.

¹ H NMR (MeOD, 250 MHz): δ [ppm] = 2.91 (s, 2H), 1.43 (s, 9H), 1.38 (s, 6H).

HPLC-MS (method A): [m/z]: 232.95 [M+H] ⁺ .

tert-Butyl N-(2-carbamothioyl-2,2-dimethylethyl)carbamate (365) ^H 2 ^N ^\©NHBoc s

Similar to general procedure 11, Lawsson's reagent (3.29 g, 8.14 mmol) and tert-butyl N-(2carbamoyl-2,2-dimethylethyl)carbamate (363) (3.2 g, 14.8 mmol) in DCM (65 ml) at room temperature for 24 h gave the title compound (1.67 g, 49%) as a white solid after purification by flash column chromatography (eluting with a 0-50% EtOAc/heptane gradient).

1 H NMR (MeOD, 250 MHz): δ [ppm] = 3.35 (s, 2H), 1.44 (s, 9H), 1.25 (s, 6H).

HPLC-MS (Method A): [m/z]: 254.95 [M+Na] ⁺ .

Ethyl 2-(2-{[(tert-butoxy)carbonyl]amino}-2-methylpropyl)-1,3-thiazole-4-carboxylate (366) o

^{e,oA xA} >-NHBoc

Similar to general procedure 1, tert-butyl N-(1-carbamothioyl-2-methylpropan-2-yl)carbamate (364) (0.91 g, 3.93 mmol), ethyl 3-bromo-2-oxopropanoate (0. 64 ml, 4.33 mmol) and CaCO ₃ (0.22 g, 2.16 mmol) in EtOH (10 ml) gave the title compound (0.285 g, 19%) as an orange oil after purification by flash column. chromatography (elution gradient 0-100% EtOAc/heptane).

1H NMR (DMSO-d6, 250 MHz): δ [ppm] = 8.42 (s, 1H), 4.29 (q, J=7.2 Hz, 2H), 3.39 (s, 2H), 1.36 (s, 9H), 1.30 (d, J=7.0 Hz, 3H), 1.20 (s, 6H).

HPLC-MS (method A): [m/z]: 329.00 [M+H] ⁺ .

Ethyl 2-(1-{[(tert-butoxy)carbonyl]amino}-2-methylpropan-2-yl)-1,3-thiazole-4-carboxylate (367)

Similar to general procedure 1, tert-butyl N-(2-carbamothioyl-2,2-dimethylethyl)carbamate (365) (1.67 g, 7.9 mmol), ethyl 3-bromo-2-oxopropanoate (1.2 ml , 7.9 mmol) and CaCO3 (0.4 g, 3.95 mmol) in EtOH (20 ml) were stirred at room temperature for 72 hours. MgSO ₄ (0.8 g) and CaCO ₃ were then added to the reaction mixture. (0.4 g, 3.95 mmol) and heated at 80°C for 7 h. The reaction mixture was cooled and the solvent was evaporated, obtaining a residue, which was dissolved in EtOAc (50 ml) and washed with saturated aqueous NaHCO ₃ (7 ml ). The aqueous layer was extracted with ethyl acetate (3x15 ml) and the combined organic layers were dried (MgSO ₄ ), filtered and evaporated to give a brown oil (2.61 g). The resulting oil was dissolved in DCM (50 ml) and TEA (2.9 ml, 20.58 mmol) was added, then di-tert-butyl dicarbonate (3.74 g, 17.15 mmol) in DCM (20 ml) was added dropwise ). The reaction mixture was stirred at room temperature for 22 hours. The reaction mixture was diluted with DCM (10 ml), the organic layer was washed with water (20 ml), brine (20 ml), dried (MgSO ₄ ), filtered and evaporated to give brown oil (4.82 g). Purification by flash column chromatography (eluting with 0-50% EtOAc-heptane) gave the title compound (1.4 g, 36%) as a yellow oil.

1H NMR (DMSO-d6, 250 MHz): δ [ppm] = 8.06 (s, 1H), 5.19 (br s, 1H), 4.40 (q, J=7.1 Hz, 2H), 3.50 (br s, 2H), 1.44 (s, 6H), 1.42 (s, 9H).

HPLC-MS (method A): [m/z]: 329.00 [M+H] ⁺ .

2-(2-{[(tert-Butoxy)carbonyl]amino}-2-methylpropyl)-1,3-thiazole-4-carboxylic acid (368)

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Similar to general procedure 5, ethyl 2-(2-{[(tert-butoxy)carbonyl]amino}-2-methylpropyl)-1,3thiazole-4-carboxylate (366) (0.285 g, 0.867 mmol, 85% pure) and LiOH (88 mg, 3.67 mmol) in THF (4 mL) and H2O (2 mL) at room temperature for 5 h gave the title compound (0.215 g, 66%, 68% pure) as dark yellow oils. The compound was used in the next step without further purification.

1H NMR (DMSO-d ₆ , 250 MHz): δ [ppm] = 8.35 (s, 1H), 3.38 (s, 2H), 1.43 (s, 9H), 1.21 (s, 6H).

HPLC-MS (method A): [m/z]: 301.00 [M+H] ⁺ .

2-(1-{[(tert-Butoxy)carbonyl]amino}-2-methylpropan-2-yl)-1,3-thiazole-4-carboxylic acid (369)

As in general procedure 5, ethyl 2-(1-{[(tert-butoxy)carbonyl]amino}-2-methylpropan-2-yl)1,3-thiazole-4-carboxylate (367) (1.4 g, 4 .12 mmol) and LiOH (0.49 g, 20.6 mmol) in THF (20 ml) and water (10 ml) at room temperature over 3 hours gave the title compound (1.5 g, 98%, 83% pure) as a yellow oil that solidified on standing. Further used without purification.

1H NMR (DMSOA, 250 MHz): δ [ppm] = 8.32 (s, 1H), 6.98-6.88 (m, 1H), 3.18 (d, J=6.5 Hz, 2H), 1.34 (s, 9H), 1.32 (s, 6H).

HPLC-MS (method A): [m/z]: 301.05 [M+H] ⁺ .

tert-Butyl N-[1-(4-{[(3-fluoropyridin-2-yl)methyl]carbamoyl}-1,3-thiazol-2-yl)-2-methylpropan-2yl]carbamate (370)

Similar to general procedure 6, 2-(2-{[(tert-butoxy)carbonyl]amino}-2-methylpropyl)-1,3-thiazole4-carboxylic acid (368) (0.28 g, 0.63 mmol, 68 % purity), (3-fluoropyridin-2-yl)methanamine dihydrochloride (A2) (0.20 g, 1.01 mmol), HATU (0.4 g, 1.04 mmol) and DIPEA (0.4 ml, 2.29 mmol) in DMF (6 mL) was stirred at room temperature for 2 h to give the title compound (0.33 g, 84%, 72% pure) as a dark yellow oil after purification by flash column chromatography ( eluting with a gradient of 0-100% EtOAc/heptane). Further used without further purification.

¹ H NMR (DMSO-d ₆ , 250 MHz): δ [ppm] = 8.58 (t, J=5.6 Hz, 1H), 8.39 (dt, J=4.7, 1 .5 Hz, 1H), 8.19 (s, 1H), 7.78-7.63 (m, 1H), 7.47-7.35 (m, 1H), 6.68 (s, 1H) , 4.67 (dd, J=5.6, 1.3 Hz, 2H), 3.41 (s, 2H), 1.41 (s, 9H), 1.23 (s, 6H).

HPLC-MS (method A): [m/z]: 409.05 [M+H]+.

tert-Butyl methylpropyl]carbamate (371)

N-[2-(4-{[(3-fluoropyridin-2-yl)methyl]carbamoyl}-1,3-thiazol-2-yl)-2-

Similar to general procedure 6, 2-(1-{[(tert-butoxy)carbonyl]amino}-2-methylpropan-2-yl)-1,3thiazole-4-carboxylic acid (369) (0.64 g, 1. 73 mmol, 83% purity), (3-fluoropyridin-2-yl)methanamine dihydrochloride (A2) (0.41 g, 2.08 mmol), HATU (0.79 g, 2.08 mmol) and DIPEA (0 .99 ml, 5.71 mmol) in DCM (8 ml) at room temperature for 17 h gave the title compound (1.78 g, 90% pure) after purification by flash column chromatography (eluting with a 0100% gradient EtOAc/heptane) as a yellow oil which solidified on standing.

1H NMR (DMSO-d ₆ , 250 MHz): δ [ppm] = 8.71 (t, J=5.8 Hz, 1H), 8.43-8.33 (m, 1H), 8 .15 (s, 1H), 7.767.65 (m, 1H), 7.47-7.34 (m, 1H), 6.97 (t, J=6.3 Hz, 1H), 4.67 ( dd, J=5.8, 1.5 Hz, 2H), 3.25 (d, J=6.5 Hz, 2H), 1.34 (s, 6H), 1.34 (s, 9H).

HPLC-MS (method A): [m/z]: 409.45 [M+H] ⁺ .

2-(2-Amino-2-methylpropyl)-N-[(3-fluoropyridin-2-yl)methyl]-1,3-thiazole-4-carboxamide dihydrochloride (372)

Similar to general procedure 2, tert-butyl N-[1-(4-{[(3-fluoropyridin-2-yl)methyl]karbamoyl}-1,3-216 039916 thiazol-2-yl)-2-methylpropan-2 -yl]carbamate (370) (0.33 g, 0.58 mmol, 72% pure) and 12M HC (0.73 ml,

8.78 mmol) in MeOH (5 ml) at 50°C for 2 h gave the title compound (0.26 g, 90%,

76% pure) as a beige residue, which was used in the next step without purification.

1H NMR (MeOD, 250 MHz): δ [ppm] = 8.53 (dd, J=5.3, 1.0 Hz, 1H), 8.24 (s, 1H), 8.16- 8.06 (m, 1H), 7.827.73 (m, 1H), 4.91 (d, J=1.4 Hz, 2H), 3.42 (s, 2H), 1.46 (s, 6H ).

HPLC-MS (method A): [m/z]: 308.95 as free base [M+H]+.

2-(1-Amino-2-methylpropan-2-yl)-N-[(3-fluoropyridin-2-yl)methyl]-1,3-thiazole-4-carboxamide dihydrochloride (373)

Similar to general procedure 2, tert-butyl N-[2-(4-{[(3-fluoropyridin-2-yl)methyl]carbamoyl}-1,3thiazol-2-yl)-2-methylpropyl]carbamate (371) ( 1.78 g, 3.93 mmol, 90% pure) and 12M HC (6.61 mL, 79.3 mmol) in MeOH (30 mL) at room temperature over 70 h gave the title compound (1.54 g, 98%) as a brown foam. Further used without purification.

1H NMR (MeOD, 250 MHz): δ [ppm] = 8.49-8.44 (m, 1H), 8.24 (s, 1H), 7.98-7.88 (m, 1H ), 7.68-7.59 (m, 1H), 4.88 (d, J=1.5 Hz, 2H), 3.37 (s, 2H), 1.58 (s, 6H).

HPLC-MS (method A): [m/z]: 309.00 as free base [M+H]+.

2-{2-[(1H-1,3-Benzodiazol-2-ylmethyl)amino]-2-methylpropyl}-H-[(3-fluoropyridin-2-yl)methyl]-1,3thiazol-4-carboxamide ( illustrative example No. 231)

Similar to general procedure 3, 2-(2-amino-2-methylpropyl)-H-[(3-fluoropyridin-2-yl)methyl]-1,3thiazol-4-carboxamide dihydrochloride (372) (0.26 g, 0 .52 mmol, 77% pure) 1H-1,3-benzodiazol-2carbaldehyde (0.106 g, 0.73 mmol), DIPEA (0.36 ml, 2.09 mmol) in MeOH (5 ml) at 50°C in over 3.5 h, after addition of NaBH ₄ (35 mg, 0.93 mmol) at 0°C, gave the title compound (0.054 g, 24%) as a cream-colored solid after purification by kp-NH column flash chromatography (elution gradient 0-6% MeOH/DCM), followed by flash column chromatography (elution gradient 0-20% MeOH/DCM).

1H NMR (MeOD, 500 MHz): δ [ppm] = 8.14-8.12 (m, 1H), 8.11 (s, 1H), 7.52-7.46 (m, 1H ), 7.46-7.41 (m, 2H), 7.27-7.22 (m, 1H), 7.17-7.12 (m, 2H), 4.70 (d, J=1 .5 Hz, 2H), 4.21(s, 2H), 3.27(s, 2H), 1.25(s, 6H).

HPLC-MS (Method C): [m/z]: 439.1 [M+H]+.

2-(1-{[2-(1H-1,3-benzodiazol-2-yl)ethyl]amino}-2-methylpropan-2-yl)-N-[(3-fluoropyridin-2yl)methyl]-1 ,3-thiazole-4-carboxamide (illustrative example No. 242)

Similar to general procedure 8, 2-(1-Amino-2-methylpropan-2-yl)-H-[(3-fluoropyridin-2-yl)methyl]1,3-thiazole-4-carboxamide dihydrochloride (373) (1 .54 g, 3.85 mmol), N-(2-nitrophenyl) prop-2-enamide (D) (0.81 g, 4.24 mmol) and DBU (1.73 ml, 11.56 mmol) in MeCN (35 ml) at room temperature for 18 h gave a mixture of mono:bis-alkylated adducts (3.7:1) (2.23 g) as an orange oil. It was further reacted with iron powder (0.86 g) in AcOH (10 mL) at 75°C for 0.5 h to give the title compound (0.29 g, 17%) as an off-white solid. after purification by flash column chromatography (elution gradient 0-40% MeOH/DCM), followed by kp-NH column chromatography (elution gradient 0-5% MeOH/DCM) and basic prep-HPLC.

1H NMR (MeOD, 500 MHz): δ [ppm] = 8.30-8.25 (m, 1H), 7.97 (s, 1H), 7.57-7.52 (m, 1H ), 7.43-7.35 (m, 2H), 7.34-7.29 (m, 1H), 7.18-7.13 (m, 2H), 4.65 (d, J=1 .5 Hz, 2H), 3.09-3.05 (m, 2H), 3.05-3.01 (m, 2H), 2.98 (s, 2H), 1.46 (s, 6H) .

HPLC-MS (method C): [m/z]: 453.1 [M+H]+.

The above general scheme 1:

N-(1H-1,3-benzodiazol-2-ylmethyl)-N-[2-(4-{[(3-methylpyridin-2-yl)methyl]carbamoyl}-1,3-thiazol2-yl)ethyl] carbamate (441)

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Similar to general procedure 6, 2-{2-[(1H-1,3-benzodiazol-2-ylmethyl)[(tert-butoxy)carbonyl]amino]ethyl}-1,3-thiazole-4-carboxylic acid (8) (2 .75 g, 6.49 mmol), 1-(3methylpyridin-2-yl)methanamine (1.19 g, 9.74 mmol), DIPEA (3.39 ml, 19.47 mmol) and HATU (4.94 g, 12.98 mmol) in DMF (50 mL) over 16 h gave the title compound (1.7 g, 51%) as a yellow foam after purification by flash column chromatography (Kp-NH, gradient elution 20100% EtOAc/heptane), followed by azeotropic drying with heptane.

¹ H NMR (MeOD, 250 MHz): δ [ppm] = 8.32 (d, J=4.4 Hz, 1H), 8.09 (s, 1H), 7.63 (d, J = 7.6 Hz, 1H), 7.53 (s, 2H), 7.30-7.14 (m, 3H), 4.74 (s, 2H), 4.69 (s, 2H), 3 .86 (s, 2H), 3.34 (s, 3H), 2.40 (s, 3H), 1.40 (s, 10H).

HPLC-MS (method C): [m/z]: 507.1 [M+H]+.

2-{2-[(1H-1,3-Benzodiazol-2-ylmethyl)amino]ethyl}-N-[(3-methylpyridin-2-yl)methyl]-1,3-thiazole-4carboxamide trihydrochloride (illustrative example No. 55)

Similar to general procedure 2, tert-butyl N-(1H-1,3-benzodiazol-2-ylmethyl)-N-[2-(4-{[(3methylpyridin-2-yl)methyl]carbamoyl}-1,3- thiazol-2-yl)ethyl]carbamate (441) (1.7 g, 3.36 mmol) and 4M HC/dioxane (8.4 ml) in dioxane (30 ml) at room temperature for 16 h gave the indicated in title compound (1.09 g, 63%) as a white solid after trituration in Et ₂ O (2x30 ml), DCM (2x20 ml) and Et ₂ O (2x30 ml), followed by recrystallization from DCM/MeOH and heptane.

1H NMR (DMSO-d6, 500 MHz): δ [ppm] = 10.29 (s, 1H), 9.56 (t, J=5.6 Hz, 1H), 8.64-8, 57 (m, 1H), 8.37 (d, J=7.7 Hz, 1H), 8.29 (s, 1H), 7.85 (dd, J=7.8, 5.8 Hz, 1H ), 7.73 (dt, J=6.6, 3.3 Hz, 2H), 7.41 (dt, J=6.1, 3.3 Hz, 2H), 4.85 (d, J= 5.7 Hz, 3H), 4.74 (s, 2H), 3.64 (dt, J=35.5, 7.0 Hz, 5H), 2.50 (s, 3H).

HPLC-MS (Method C): [m/z]: 407.05 [M+H] ⁺ .

The above general scheme 41:

tert-butyl 3-[(pyridin-2-ylmethyl)carbamoyl]-4H,5H,6H,7H-thieno[2,3-c]pyridine-6-carboxylate (449)

Similar to the general procedure 6, 6-(tert-Butoxycarbonyl)-4,5,6,7-tetrahydrothieno[2,3-c]pyridine-3-carboxylic acid (220 mg, 0.77 mmol), 1-(pyridine-2- yl)methanamine (88 µl, 0.85 mmol), DIPEA (407 µl, 2.33 mmol) and HATU (443 mg, 1.17 mmol) in DCM (12 ml) at room temperature for 1 h gave the indicated in title compound (466 mg) as a yellow oil after purification by flash column chromatography (eluting with a 50-100% EtOAc/heptane gradient).

1H NMR (CDCl ₃ , 500 MHz): δ [ppm] = 8.55 (d, J=4.8 Hz, 1H), 7.68 (td, J=7.7, 1.8 Hz , 1H), 7.64 (s, 1H), 7.31 (d, J=7.8 Hz, 1H), 7.22 (dd, J=7.2, 5.2 Hz, 1H), 4 .70 (d, J=4.8 Hz, 2H), 4.62 (s, 2H), 3.76-3.61 (m, 2H), 2.99 (s, 2H), 1.48 ( s, 9H), 1.44 (d, J=6.6 Hz, 1H).

HPLC-MS (Method E): [m/z]: 374.05 [M+H] ⁺ .

N-(pyridin-2-ylmethyl)-4H,5H,6H,7H-thieno[2,3-c]pyridine-3-carboxamide (450)

As in general procedure 2, t-butyl 3-[(pyridin-2-ylmethyl)carbamoyl]-4H,5H,6H,7Hthieno[2,3-c]pyridine-6-carboxylate (449) (466 mg, 1.09 mmol) and TFA (940 μl) in DCM (5 ml) at room temperature for 16 hours gave the title compound (130 mg, 44%) as a yellow oil. The compound was used in the next step without purification.

1H NMR (MeOD, 500 MHz): δ [ppm] = 8.49 (d, J=4.3 Hz, 1H), 7.92 (s, 1H), 7.82 (td, J= 7.7, 1.8 Hz, 1H), 7.43 (d, J=7.9 Hz, 1H), 7.37-7.27 (m, 1H), 4.59 (s, 2H), 4.16 (s, 2H), 3.21 (t, J=6.0 Hz, 2H), 3.01 (t, J=6.0 Hz, 2H).

HPLC-MS (Method E): [m/z]: 273.95 [M+H] ⁺ .

6-(1H-1,3-Benzodiazol-2-ylmethyl)-N-(pyridin-2-ylmethyl)-4H,5H,6H,7H-thieno[2,3-c]pyridine-3carboxamide (Illustrative Example No. 48 )

- 218 039916

Similar to general procedure 7, N-(pyridin-2-ylmethyl)-4H,5H,6H,7H-thieno[2,3-c]pyridine-3carboxamide (450) (65 mg, 0.24 mmol), K2CO3 (49 mg, 0.36 mmol) and 2-(chloromethyl)-1H-benzimidazole (44 mg, 0.26 mmol) in acetone (3 mL) at room temperature over 72 h gave the title compound (25 mg, 26% ) as a pale yellow solid after purification by flash column chromatography (kp-NH, 0-10% MeOH/DCM gradient elution), followed by basic prep-HPLC.

1H NMR (DMSO-d ₆ , 500 MHz): δ [ppm] = 12.36 (s, 1H), 8.78 (t, J=6.0 Hz, 1H), 8.50 (d , J=4.7 Hz, 1H), 7.95 (s, 1H), 7.76 (td, J=7.7, 1.7 Hz, 1H), 7.49 (s, 2H), 7 .31 (d, J=7.8 Hz, 1H), 7.29-7.23 (m, 1H), 7.14 (dd, J=5.9, 3.1 Hz, 2H), 4, 49 (d, J=6.0 Hz, 2H), 3.93 (s, 2H), 3.73 (s, 2H), 2.89 (d, J=5.5 Hz, 2H), 2, 79 (t, J=5.7 Hz, 2H).

HPLC-MS (Method D): [m/z]: 404.2 [M+H]+.

6-[2-(1H-1,3-Benzodiazol-2-yl)ethyl]-N-(pyridin-2-yl-methyl)-4H,5H,6H,7H-thieno [2,3-c]pyridine -3carboxamide (illustrative example No. 49)

Similar to general procedure 7, N-(pyridin-2-ylmethyl)-4H,5H,6H,7H-thieno[2,3-c]pyridine-3carboxamide (450) (65 mg, 0.24 mmol), K2CO3 (49 mg, 0.36 mmol) and 2-(2-chloroethyl)-1H-benzimidazole (47 mg, 0.26 mmol) in acetone (3 ml) at room temperature for 24 hours, after adding DMF (5 ml), NaI (39 mg, 0.26 mmol), DIPEA (0.16 ml, 0.95 mmol) and 2-(2-chloroethyl)-1H-benzimidazole (94 mg, 0.52 mmol), at room temperature for 72 h gave the title compound (9 mg, 9%) as an orange solid after purification by basic prep-HPLC.

¹ H NMR (DMSO-d ₆ , 500 MHz): δ [ppm] = 12.17 (s, 1H), 8.75 (t, J=6.0 Hz, 1H), 8.50 ( d, J=4.2 Hz, 1H), 7.94 (s, 1H), 7.75 (td, J=7.7, 1.8 Hz, 1H), 7.46 (s, 2H), 7.31 (d, J=7.8 Hz, 1H), 7.28-7.23 (m, 1H), 7.10 (dd, J=6.0, 3.1 Hz, 2H), 4 .48 (d, J=6.0 Hz, 2H), 3.71 (s, 2H), 3.05 (t, J=7.2 Hz, 2H), 2.96 (t, J=7, 2 Hz, 2H), 2.82 (d, J=5.3 Hz, 2H), 2.75 (t, J=5.7 Hz, 2H).

HPLC-MS (method B): [m/z]: 418.2 [M+H]+.

The above general scheme 42:

2-{2-[N-(1H-1,3-benzodiazol-2-ylmethyl)-2-bromoacetamido]ethyl}-N-[(3-fluoropyridin-2-yl)methyl]1,3-thiazol-4 -carboxamide (451)

2-{2-[(1H-1,3-Benzodiazol-2-ylmethyl)amino]ethyl}-N-[(3-fluoropyridin-2-yl)methyl]-1,3-thiazole-4carboxamide (Illustrative Example No. 40) (150 mg, 0.37 mmol) and TEA (127 μl, 0.91 mmol) were dissolved in DCM (10 ml) and bromoacetyl chloride (61 μl, 0.73 mmol) was added dropwise. The resulting mixture was stirred at room temperature for 15 min, then quenched with saturated aqueous NaHCO ₃ (20 ml) and extracted with DCM (3x30 ml) and 4:1 chloroform/IPS (30 ml). The combined organic extracts were dried (Na2SO4) and evaporated in vacuo. Purification by flash column chromatography (eluting with a 0-5% MeOH/DCM gradient) gave the title compound (224 mg, 82% pure) as a brown residue. The compound was used in the next step without further purification.

HPLC-MS (method M): [m/z]: 530.85/532.85 [M+H]+.

N-[(3-fluoropyridin-2-yl)methyl]-2-(2-{12-oxo-1,8,11-triazatricyclo[7.4.0.0 ² ' ⁷ ]trideca-2(7) ,3,5,8tetraen-11-yl}ethyl)-1,3-thiazole-4-carboxamide (illustrative example No. 275)

2-{2-[Na-(III-1,3-benzodiazol-2-ylmethyl)-2-bromoacetamido]ethyl}-Na-[(3-fluoropyridin-2-yl)methyl]1,3-thiazol-4 -carboxamide (451) (220 mg, 0.41 mmol, 82% purity) was dissolved in THF (10 ml) and NaH (60%, 50 mg, 1.24 mmol) was added. The resulting mixture was stirred at room temperature for 10 minutes. The reaction was quenched with saturated aqueous NaHCO ₃ (20 ml) and was extracted with DCM (3x20 ml) and a mixture of 4:1 chloroform/IPS (20 ml). The combined organic extracts were dried (Na ₂ SO ₄ ) and

- 219 039916 was evaporated in vacuum. Purification by basic prep-HPLC gave the title compound (15 mg, 8%) as an off-white solid.

¹ H NMR (DMSO-d ₆ , 500 MHz): δ [ppm] = 8.65 (t, J=5.6 Hz, 1H), 8.36-8.32 (m, 1H), 8.18 (s, 1H), 7.717.64 (m, 1H), 7.61 (dt, J=5.4, 3.4 Hz, 1H), 7.56-7.51 (m, 1H) , 7.40-7.34 (m, 1H), 7.26-7.21 (m, 2H), 4.91 (s, 2H), 4.88 (s, 2H), 4.63-4 .57 (m, 2H), 3.93 (t, J=7.2 Hz, 2H), 3.42 (t, J=7.2 Hz, 2H).

HPLC-MS (method B): [m/z]: 451.2 [M+H]+.

N-[(3-fluoropyridin-2-yl)methyl]-2-(2-{1,8,11-triazatricyclo[7.4.0.0 ^2.7 ]trideca-2(7),3.5 ,8-tetraen11-yl}ethyl)-1,3-thiazole-4-carboxamide (illustrative example No. 276)

2-{2-[(1H-1,3-Benzodiazol-2-ylmethyl)amino]ethyl}-N-[(3-fluoropyridin-2-yl)methyl]-1,3-thiazole-4carboxamide (illustrative example no. 40) (150 mg, 0.37 mmol) and TEA (509 μl, 3.65 mmol) were combined in DMF (4 ml) and 1,2-dibromoethane (315 μl, 3.65 mmol) was added. The resulting mixture was heated at 100°C for 40 min, then cooled to room temperature and quenched with saturated aqueous NaHCO ₃ . The resulting mixture was extracted with dichloromethane (3x50 ml), the combined organic extracts were washed with an aqueous solution of LiCl (2M, 50 ml), dried (Na ₂ SO ₄ ) and evaporated in vacuum. Purification by basic prep-HPLC followed by flash column chromatography (elution with a 0-4% MeOH/DCM gradient) gave the title compound (45 mg, 28%) as a pale yellow solid.

¹ H NMR (MeOD, 500 MHz): δ [ppm] = 8.37 (dt, J=4.7, 1.2 Hz, 1H), 8.09 (s, 1H), 7.64 -7.57(m, 2H), 7.517.46(m, 1H), 7.42-7.37(m, 1H), 7.32-7.26(m, 2H), 4.80(d , J=1.6 Hz, 2H), 4.23 (t, J=5.5 Hz, 2H), 4.04 (s, 2H), 3.41 (t, J=6.8 Hz, 2H ), 3.25-3.20 (m, 2H), 3.16 (t, J=6.8 Hz, 2H).

HPLC-MS (method B): [m/z]: 437.1 [M+H] ⁺ .

tert-Butyl (2-(4-(((3-fluoropyridin-2-yl)methyl)carbamoyl)-1H-1,2,3-triazol-1-yl)ethyl)carbamate (452)

1-(2-((tert-butoxycarbonyl)amino)ethyl)-1H-1,2,3-triazole-4-carboxylic acid (commercially available) (2.0 g, 7.8 mmol) in 50 ml DCM was cooled to -5°C and added to the resulting mixture of N-methylmorpholine (0.94 ml, 8.6 mmol) and ethyl chloroformate. The temperature was kept below 0°C during the addition, and at the end of the addition the mixture was stirred at 0°C for 1 hour. A second portion of N-methylmorpholine (2.7 ml, 24.2 mmol) and (3-Fluoropyridin-2-yl) methanamine dihydrochloride (A2). The reaction mixture was allowed to warm to room temperature and stirred overnight. Was added to the reaction mixture 50 ml of water and was extracted with DCM. The organic phase was dried (Na ₂ SO ₄ ), filtered and evaporated in vacuo to give a crude product which was purified by flash column chromatography to give the title compound (2.1 g, 74%).

¹ H NMR (DMSO, 400 MHz): δ [ppm] = 8.82 (t, 1H), 8.50 (s, 1H), 8.38 (d, 1H), 7.70 (m , 1H), 7.40 (m, 1H), 7.01 (s, 1H), 4.65 (d, 2H), 4.46 (m, 2H), 3.39 (m, 2H), 1 .40-1.24 (m, 9H).

-(2-Aminoethyl)-N-((3-fluoropyridin-2-yl)methyl)- 1H-1,2,3-triazole-4-carboxamide hydrochloride (453) ml conc. HC was added to a solution of tert-butyl (2-(4-(((3-fluoropyridin-2-yl)methyl)carbamoyl)-1H1,2,3-triazol-1-yl)ethyl)carbamate (452) (2, 00 g, 5.49 mmol) in 100 ml methanol and stirred at room temperature for 2 h. The resulting mixture was evaporated in vacuo to give the title compound (1.74 g, 99%) as a white solid.

¹ H NMR (DMSO, 400 MHz): δ [ppm] = 8.99 (t, 1H), 8.73 (s, 1H), 8.51 (s, 3H), 8.45 (d , 1H), 7.87 (m, 1H), 7.53 (m, 1H), 4.78 (m, 2H), 4.71 (d, 2H), 3.37 (m, 2H).

N-((3-Fluoropyridin-2-yl)methyl)-1 -(2-((3-((2-nitrophenyl)amino)-3-oxopropyl)amino)ethyl)- 1H1,2,3-triazole- 4-carboxamide (454)

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1-(2-aminoethyl)-N-((3-fluoropyridin-2-yl)methyl)-1H-1,2,3-triazole-4-carboxamide hydrochloride (453) (4.46 g, 13.2 mmol ) and DBU (10.1 g, 66.1 mmol) were suspended in 120 ml of acetonitrile. N-(2-nitrophenyl)prop-2-enamide (D) (2.54 g, 13.2 mmol) in 10 ml of acetonitrile was added dropwise to the reaction mixture over 1 h and stirred at room temperature overnight. The reaction mixture was evaporated to dryness and redissolved in 100 ml DCM. 50 ml water was added and extracted with DCM. The organic phases were dried (Na ₂ SO ₄ ), filtered and evaporated in vacuo to give a crude product which was purified by flash column chromatography to give the title compound (2.1 g, 35%).

1H NMR (DMSO, 400 MHz): δ [ppm] = 10.55 (s, 1H), 8.78 (t, 1H), 8.57 (s, 1H), 8.38 (d, 1H), 7.93(m, 1H), 7.76-7.62(m, 3H), 7.40(m, 1H), 7.32(m, 1H), 4.65(d, 2H ), 4.50 (m, 2H), 3.02 (t, 2H), 2.81 (t, 2H), 2.51 (m, 2H).

1-(2-((2-(1H-benzo[d]imidazol-2-yl)ethyl)amino)ethyl)-N-((3-fluoropyridin-2-yl)methyl)-1H-1,2, 3-triazole-4-carboxamide (illustrative example No. 277)

N-((3-fluoropyridin-2-yl)methyl)-1 -(2-((3 -((2-nitrophenyl)amino)-3-oxopropyl)amino)ethyl)-1 H1,2,3-triazole -4-carboxamide (454) (900 mg, 1.97 mmol) and iron powder (335 mg, 5.92 mmol) were suspended in 10 ml of acetic acid and stirred under nitrogen at 80° C. for 2 hours. evaporated to dryness, and the resulting residue was added 60 ml of a mixture of chloroform/isopropanol (1:4). The pH was adjusted to 11 by adding 1N. NaOH solution, and the phases were separated. After extraction of the aqueous phase with chloroform, the combined organic phases were dried (Na ₂ SO ₄ ), filtered and evaporated in vacuo to give a crude product which was purified by flash column chromatography to give the title compound (430 mg, 53%).

¹ H NMR (H4-methanol, 400 MHz): δ [ppm] = 8.40 (s, 1H), 8.38 (m, 1H), 7.63-7.58 (m, 1H) , 7.50-7.45(m, 2H), 7.41-7.36(m, 1H), 7.18-12(m, 2H), 4.78(d, 2H), 4.56 (m, 2H), 3.16 (t, 2H), 3.10-3.0 (m, 4H).

5-(2-((tert-Butoxycarbonyl)amino)ethyl)-1,2,4-oxadiazole-3-carboxylic acid

Ethyl 5-(2-((tert-butoxycarbonyl)amino)ethyl)-1,2,4-oxadiazole 3-carboxylate (4 g, 14.06 mmol), THF (80 ml) and water (20 ml ) with stirring at room temperature. Lithium hydroxide (1 g, 41.75 mmol) was then added to the solution and the reaction mixture was stirred at room temperature for 2 h. The reaction mixture was acidified to pH 2.8 (with 32% aqueous HC), extracted with ethyl acetate ( 3x80 ml), dried (Na ₂ SO ₄ ), filtered and evaporated to give the title compound as a brown oil (2.95 g) which was used without purification.

tert-Butyl (2-(3-((pyridin-2-ylmethyl)carbamoyl)-1,3,4-oxadiazol-5-yl)ethyl)carbamate

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To a solution of 5-(2-((tert-butoxycarbonyl)amino)ethyl)-1,2,4-oxadiazole-3-carboxylic acid (2.95 g, 11.47 mmol) in 100 ml of dichloromethane at 0°C was added 4-methylmorpholine (2.8 ml, 25.47 mmol) and ethyl chloroformate (1 ml, 10.46 mmol). After 60 min stirring at 0° C., 2-picolylamine (1.7 mL, 16.82) was added and the reaction mixture was stirred 2 h at room temperature. The reaction mixture was washed with water (100 ml), dried (Na ₂ SO ₄ ), filtered and evaporated. The crude product was purified using a 100 g SNAP Ultra column (eluting with a gradient of 0-100% ethyl acetate in petroleum ether) to give a pale brown oil (2.29 g, 6.6 mmol, 58%).

1H NMR (400 MHz, CDCl ₃ ): δ 8.5 (s, 1H), 8.1 (m, 2H), 7.7 (m, 1H), 7.3 (m, 2H), 6.8 (s, 1H), 4.6 (s, 2H), 3.4 (m, 2H), 2.5 (m, 2H), 1.4 (s, 9H).

5-(2-Aminoethyl)-N-(pyridin-2-ylmethyl)-1,2,4-oxadiazole-3-carboxamide dihydrochloride

HCl

5-(2-aminoethyl)-M-(pyridin-2-ylmethyl)-1,2,4-oxadiazole-3-carboxamide dihydrochloride

To a solution of tert-butyl (2-(3-((pyridin-2-ylmethyl)carbamoyl)-1,2,4-oxadiazol-5-yl)ethyl)carbamate (2.41 g, 6.94 mmol) in dioxane (25 ml) was added a 4M solution of HC in dioxane (25 ml). After stirring at room temperature for 2 hours, the reaction mixture was evaporated to give the title compound as a brown oil (2.83 g) which was used without purification.

5-(2-((3-((2-nitrophenyl)amino)-3-oxopropyl)amino)ethyl)-N-(pyridin-2-ylmethyl)-1,2,4oxadiazole-3-carboxamide

O

Ν ^N Y ⁺ DBU

HCI θ ^V -NH _a

HCl

Η . N about

acetonitrile

Η,Ν

Η •Ν.

Ο,Ν Η .Ν

5-(2-((3-((2-nitrophenyl)amino)-3-oxopropyl)amino)ethyl)-N-(pyridin-2-ylmethyl)-1,2,4-oxadiazole-3carboxamide

In a round bottom flask, 5-(2-aminoethyl)-N-(pyridin-2-ylmethyl)-1,2,4-oxadiazole-3-carboxamide dihydrochloride (3.48 g, 9.76 mmol) was suspended in acetonitrile (100 ml ) under nitrogen, 1,8-diazabicycloundec-7-ene (DBU) (7.1 mL, 47.48 mmol, 4.9 eq.) was added dropwise, followed by N-(2nitrophenyl)prop-2-enamide ( 1.9 g, 9.89 mmol). The reaction mixture was stirred at room temperature overnight, then evaporated, diluted with 100 ml dichloromethane, washed with 50 ml water, dried (Na ₂ SO ₄ ), filtered and evaporated. The crude product was purified using a 100 g SNAP Ultra column (elution with a gradient of 0-25% methanol in dichloromethane) to give a light brown oil (0.74 g, 1.7 mmol, 17%).

1H NMR (400 MHz, CDCl ₃ ): δ 8.5 (s, 1H), 8.1 (m, 2H), 7.8 (m, 1H), 7.6 (m, 1H), 7.3 (m, 3H), 4.4 (s, 2H), 3.1 (m, 4H), 2.7 (m, 2H), 2.5 (m, 2H).

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5-(2-((2-(1H-benzo[d]imidazol-2-yl)ethyl)amino)ethyl)-N-(pyridin-2-ylmethyl)-1,2,4-oxadiazole-3carboxamide (illustrative example no. 278)

ASON

Fe---------1

about

5-(2-((2-(1H-benzo[c1]imidazol-2-yl)ethyl)amino)ethyl)-U-(pyridin-2-ylmethyl)-1,2,4-oxadiazole-3carboxamide

5-(2-((3-((2-Nitrophenyl)amino)-3-oxopropyl)amino)ethyl)-N-(pyridin-2-ylmethyl)-1,2,4oxadiazole-3-carboxamide (0.74 g, 1.68 mmol) was dissolved in acetic acid (5 ml) under nitrogen atmosphere and iron powder (0.3 g) was added. The reaction mixture was heated at 80° C. for 3 hours. After cooling to room temperature, toluene (50 ml) was added and the resulting mixture was evaporated. A mixture of chloroform: IPA (4:1, 40 ml) was added, the reaction mixture was cooled to 0°C, basified with 6M NaOH solution to pH 10-12, filtered, dried (Na ₂ SO ₄ ), filtered and evaporated. The crude product was purified using a 50 g SNAP KP-Sil column (elution with a gradient of 0-25% methanol in dichloromethane) to give a light brown oil (0.31 g, 0.8 mmol, 47%).

1H NMR (400 MHz, MeOD): δ 8.4 (s, 1H), 7.9 (s, 1H), 7.7 (m, 2H), 7.3 (m, 1H), 7.2 ( m, 1H), 7.1 (m, 2H), 4.4 (s, 2H), 3.1 (m, 4H), 2.9 (m, 2H), 2.5 (m, 2H).

tert-Butyl (2-(3-(((3-fluoropyridin-2-yl)methyl)carbamoyl)-1,2,4-oxadiazol-5-yl)ethyl)carbamate

tert-butyl (2-(3-(((3-fluoropyridin-2-yl)methyl)carbamoyl)-1,2,4-oxadiazol-5-yl)ethyl)carbamate

To a solution of 5-(2-((tert-butoxycarbonyl)amino)ethyl)-1,2,4-oxadiazole-3-carboxylic acid (8.16 g, 31.72 mmol) in 400 ml dichloromethane at -5°C 4-methylmorpholine (20 ml, 182 mmol) and ethyl chloroformate (2.7 ml, 28.24 mmol) were added. After 60 minutes of stirring at -5° C., (3-fluoropyridin2-yl)methanamine dihydrochloride (9.50 g, 47.73 mmol) was added and the reaction mixture was stirred for 2 hours at room temperature. The reaction mixture was washed with water (400 ml), dried (Na ₂ SO ₄ ), filtered and evaporated. The crude product was purified using a 100 g SNAP Ultra column (gradient elution 0-100% ethyl acetate in petroleum ether) to give a light brown oil (4.15 g, 11.4 mmol, 36%).

1H NMR (400 MHz, CDCl ₃ ): δ 8.3 (s, 1H), 7.4 (m, 1H), 7.3 (m, 1H), 6.9 (s, 1H), 4.6 (s, 2H), 3.4 (m, 2H), 2.5 (m, 2H), 1.4 (s, 9H).

5-(2-Aminoethyl)-N-((3-fluoropyridin-2-yl)methyl)-1,2,4-oxadiazole-3-carboxamide dihydrochloride

- 223 039916

5-(2-aminoethyl)-M-((3-fluoropyridin-2-yl)methyl)-1,2,4-oxadiazole-3-carboxamide dihydrochloride

To a solution of tert-butyl (2-(3-(((3-fluoropyridin-2-yl)methyl)carbamoyl)-1,2,4-oxadiazol-5yl)ethyl)carbamate (4.23 g, 11.58 mmol ) in dioxane (50 ml) was added 4M HCl solution in dioxane (50 ml). After stirring at room temperature for 3 hours, the reaction mixture was evaporated to give the title compound as a brown oil (3.78 g) which was used without purification.

N-((3-fluoropyridin-2-yl)methyl)-5-(2-((3-((2-nitrophenyl)amino)-3-oxopropyl)amino)ethyl)-1,2,4oxadiazole-3- carboxamide

N-((3-fluoropyridin-2-yl)methyl)-5-(2-((3-((2-nitrophenyl)amino)-3-oxopropyl)amino)ethyl)-1,2,4 oxadiazole-3 -carboxamide

5-(2-Aminoethyl)-N-((3-fluoropyridine-2-yl)methyl)-1,2,4oxadiazole-3-carboxamide dihydrochloride (3.72 g, 9.93 mmol) in acetonitrile was suspended in a round bottom flask. (100 ml) under nitrogen and 1,8-diazabicycloundec-7-ene (DBU) (7.42 ml, 49.65 mmol, 5 eq.) was added dropwise followed by N-(2nitrophenyl)prop-2-enamide (1.91 g, 9.93). The reaction mixture was stirred at room temperature overnight, then evaporated, diluted with 100 ml dichloromethane, washed with 50 ml water, dried (Na ₂ SO ₄ ), filtered and evaporated. The crude product was purified using a 100 g SNAP KP-Sil column (elution with a gradient of 0-25% methanol in dichloromethane) to give a light brown oil (1.7 g, 3.7 mmol, 37%).

¹ H NMR (400 MHz, MeOD): δ 8.3 (s, 1H), 8.1 (m, 2H), 7.6 (m, 1H), 7.5 (m, 1H), 7.3 (m, 2H), 4.5 (s, 2H), 3.0 (m, 4H), 2.7 (m, 2H), 2.5 (m, 2H).

5-(2-((2-(1H-benzo[d]imidazol-2-yl)ethyl)amino)ethyl)-N-((3-fluoropyridin-2-yl)methyl)-1,2,4oxadiazol- 3-carboxamide (illustrative example No. 279)

5-(2-((2-(1H-benzo[c1]imidazol-2-yl)ethyl)amino)ethyl)-M-((3-fluoropyridin-2-yl)methyl)-1,2,4- oxadiazole-Zcarboxamide

N-((3-fluoropyridin-2-yl)methyl)-5-(2-((3-((2-nitrophenyl)amino)-3-oxopropyl)amino)ethyl)-1,2,4oxadiazol-3- carboxamide (2.3 g, 5.03 mmol) was dissolved in acetic acid (20 ml) under nitrogen atmosphere and iron powder (1.4 g) was added. The reaction mixture was heated at 80° C. for 3 hours. After cooling to room temperature, toluene (200 ml) was added and the resulting mixture was evaporated. A mixture of chloroform: IPA (4:1, 100 ml) was added, the reaction mixture was cooled to 0°C, basified with 6M NaOH solution to pH 10-12, filtered, dried (Na ₂ SO ₄ ), filtered and evaporated. The crude product was purified using a 100 g SNAP KP-Sil column (elution with a gradient of 0-25% methanol in dichloromethane) to give a light brown oil (1.51 g, 3.7 mmol, 74%).

- 224 039916

Ή NMR (400 MHz, MeOD): δ 8.2 (s, 1H), 7.5 (m, 3H), 7.3 (m, 1H), 7.1 (m, 2H), 4.5 ( s, 2H), 3.1 (m, 4H),

2.9 (m, 2H), 2.4 (m, 2H).

2-(1-((1H-benzo[d]imidazol-2-yl)methyl)azetidin-3-yl)-N-((3-fluoropyridin-2-yl)methyl)oxazole-5carboxamide (illustrative example No. 280 )

2-(1-((1Н-benzo[0]imidazol-2-yl)methyl)azetidin-3-yl)-MD(3-fluoropyridin-2-yl)methyl)oxazole-5carboxamide

In a round bottom flask, 2-(azetidin-3-yl)-NX(3-fluoropyridin-2-yl)methyl]-1,3-oxazol-5-carboxamide dihydrochloride (starting compound similar to that used in Illustrative Example No. 2 described above) (4 g, 11.5 mmol) was diluted with dichloromethane (1600 ml) under nitrogen atmosphere, and diisopropylethylamine (20 ml, 115 mmol, 11 eq.) was added dropwise. The reaction mixture was stirred for 10 min at room temperature and then 1H-benzo[b]imidazole-2-carbaldehyde (2 g, 13.7 mmol, 1.3 eq.) was added. The reaction mixture was stirred at room temperature for 18 hours, then sodium triacetoxyborohydride (5 g, 23.6 mmol, 2.3 eq.) was added and the reaction mixture was stirred at room temperature for 18 hours. The reaction mixture was washed with water (1000 ml), separated, dried (MgSO ₄ ), filtered and evaporated. The crude product was purified using a 100 g SNAP Ultra column followed by a 55 g SNAP NH column (elution each time with a gradient of 0-25% methanol in dichloromethane) to give a light brown oil (1.34 g, 3.3 mmol, 29% ).

1H NMR (400 MHz, CDCl ₃ ): δ 7.99 (m, 1H), 7.51 (m, 1H), 7.15 (m, 2H), 7.01 (m, 1H), 6.88 (m, 1H), 6.81 (m, 2H), 4.4 (s, 2H), 3.6 (2H), 3.42 (m, 1H), 3.35 (m, 2H), 3 .28 (m, 2H).

MS (ESI+): m/z 407 [M+H] ⁺ .

Claims (15)

CLAIM 1. A compound having the formula (A-II) o (A-H) where R ⁴ means 0, 1, 2 or 3 substituents independently selected from the group consisting of the following representatives: a halogen atom, a cyano group, a linear or branched Cl-C ₃ -alkyl bearing 0, 1, 2 or 3 halogen atoms, a C 1 -C ₂ alkoxy group and a carboxyl group; Xi, X ₂ , X ₃ and X ₄ are selected from C, N, S or O to form a group selected from: - 225 039916 where * indicates the point of attachment to the aminocarbonyl group, and ** indicates the point of attachment to the A ₁ group; and where X1, X ₃ and X ₄ when they represent C or N, are unsubstituted or carry an additional substituent selected from halogen, C ₁ -C ₃ -alkyl bearing 0, 1, 2 or 3 halogen atoms; and where Y ¹ represents a hydrogen atom, halogen or C ₁ -C ₃ -alkyl, which carries 0, 1, 2 or 3 halogen atoms; R ¹ is selected from the group consisting of a hydrogen atom and linear or branched C ₁ -C ₃ -alkyl; Cycl is selected from the group consisting of substituted 6-membered monocyclic aryl, where the aryl substituents are selected from the group consisting of F and Cl, cyano, linear or branched C ₁ -C ₃ alkyl which carries 0, 1, 2 or 3 halogen atoms, dimethylamino, methoxy, difluoromethoxy and trifluoromethoxy, pyrrolinyl, morpholinyl and piperazinyl , substituted with 0 or 1 methyl group, and a sulfonyl group of the formula substituted or unsubstituted 5- or 6-membered monocyclic heteroaryl containing 1-3 heteroatoms selected from S, O and N, where possible substituents are selected from a halogen atom, cyano, unsubstituted C ₁ -C ₃ -alkyl or C ₁ -C ₃ -alkyl, which carries 0, 1, 2 or 3 halogen atoms or hydroxy groups, C ₁ -C ₃ -alkoxy, oxo groups (=O), forming an oxo-substituted heteroaryl of the formula , N-morpholinyl groups, NH ₂ -(C=O)- aminocarbonyl groups, or dimethylamino groups; Q is a hydrogen atom or C1-C4 alkyl which can form a conjugated 5- or 6-membered cycle with Cycl; n is 0 or an integer from 1 to 2; A1 is an unsubstituted linear C1-C4 alkanediyl; And ₂ represents a linear or branched unsubstituted or substituted C1-C4-alkanediyl, where possible substituents are selected from 1 or 2 halogens, and oxo groups or a direct connection; R ³ represents a hydrogen atom or unsubstituted or substituted linear C ₁ -C ₃ -alkyl, where possible substituents are selected from the benzimidazolyl group, the cyclopropyl group and the amino NH ₂ group; or A1 and R ³ together with the nitrogen atom to which they are attached form an unsubstituted or substituted 4-6 membered mono- or bicyclic ring, where possible substituents are selected from methyl, - 226 039916 ethyl and hydroxyl group; or R ³ and A ₂ together with the nitrogen atom to which they are attached form an unsubstituted or substituted 4-membered ring, where the possible substituents are selected from a heteroaryl containing from 1 to 3 heteroatoms selected from S, O and N, and an oxo group ; or its pharmaceutically acceptable salts. 2. The compound according to claim 1, where Cycl is substituted forming compounds of formula (A-IIIb) or unsubstituted pyridinyl, or formula (A-IVb) where R ⁵ means 0, 1, 2 or 3 substituents, which are independently selected from the group consisting of the following representatives: F and Cl, methyl, trifluoromethyl, hydroxymethyl, methoxy group, oxo group (=O) forming a substituted pyridinyl group having the general formula dimethylamino group, NH ₂ -(C=O)-, a cyano group and a morpholinyl group, or pharmaceutically acceptable salts thereof. 3. The compound according to claim 1 or 2, having the formula (A-IVd) or its pharmaceutically acceptable salts. 4. The compound according to claim 2 or 3, where R ⁵ represents a halogen atom, or its pharmaceutically acceptable salts. 5. The compound of claim 1 wherein Cycl is a substituted phenyl group, forming compounds of formula (A-Vb) where R ⁶ means 1, 2 or 3 substituents, which are independently selected from the group consisting of the following representatives: F and Cl, cyano group, linear or branched C ₁ -C ₃ -alkyl bearing 0, 1, 2 or 3 halogen atoms, dimethylamino group, - 227 039916 methoxy, difluoromethoxy and trifluoromethoxy group, pyrrolinyl, morpholinyl and piperazinyl substituted with 0 or 1 methyl group and a sulfonyl group of the formula 0=3=0 I or pharmaceutically acceptable salts thereof. 6. A compound according to any one of the preceding claims 1 to 5, wherein X ₁ is O or S, X2 is C, X ₃ is N and a hydrogen atom, a halogen or a linear or branched C ₁ -C ₃ -alkyl which carries 0, 1, 2 or 3 halogen atoms; and where * indicates the point of attachment to the aminocarbonyl group, and ** indicates the point of attachment to the A ₁ group; or its pharmaceutically acceptable salts. 7. A compound according to any one of claims 1 to 6, wherein the additional substituents X1, X ₃ and X ₄ are selected from the group consisting of Cl, a methyl group, an isopropyl group, a CF ₃ group, or a pharmaceutically acceptable salt thereof. 8. A compound according to any one of claims 1 to 7, wherein n is 1 and/or Q is H and/or R ¹ is H, or pharmaceutically acceptable salts thereof. 9. Connection according to any one of claims 1 to 8, where A1 and A2 are the same or different and are independently selected from methylene and ethane-1,2-diyl, or A ₁ and R ³ together with the nitrogen atom to which they are attached form an unsubstituted or substituted 4-membered aliphatic monocyclic ring, where possible substituents are selected from methyl and hydroxyl group; or its pharmaceutically acceptable salts. 10. A compound according to any one of the preceding claims, selected from - 228 039916 - 229 039916 - 230 039916 - 231 039916 - 232 039916 - 233 039916 - 234 039916 - 235 039916 - 236 039916 - 237 039916 - 238 039916 - 239 039916 - 240 039916 - 241 039916 - 242 039916 - 243 039916 - 244 039916 or its pharmaceutically acceptable salts. 11. A compound according to any one of claims 1 to 10 having the formula or its pharmaceutically acceptable salts. 12. The use of a compound according to any one of claims 1 to 11 as a drug effective as an inhibitor of ferroportin or in inhibition of iron transport mediated by ferroportin for the prevention and/or treatment of diseases associated with or caused by a reduced level of hepcidin, or metabolic disorders iron, leading to elevated iron levels or increased iron absorption and/or iron overload. 13. Use according to claim 12, wherein the diseases associated with or caused by elevated iron levels, increased iron absorption or iron overload are selected from thalassemia, including alpha thalassemia, beta thalassemia and delta thalassemia, hemoglobinopathies, hemoglobin E disease, hemoglobin diseases H, hemochromatosis, hemolytic anemia, including sickle cell anemia and congenital dyserythropoietic anemia. 14. The use of a compound according to any one of claims 1 to 11 as a drug effective as an inhibitor of ferroportin or in inhibition of iron transport mediated by ferroportin for the prevention and/or treatment of diseases caused by a reduced level of hepcidin. 15. The use of a compound according to any one of claims 1 to 11 as a drug, effective - 245 039916 as a ferroportin inhibitor or in the inhibition of iron transport mediated by ferroportin in order to prevent and/or treat the formation of radicals, reactive oxygen species (ROS) and oxidative stress; and/or damage to the heart, liver, and endocrine system caused by iron overload; and/or inflammation initiated by excess iron.