RU2803084C1 - Aminopyridine derivatives and their use as selective alk-2 inhibitors - Google Patents

Abstract

FIELD: pharmaceutical.

SUBSTANCE: invention relates to a compound that is a hydrate of 2-amino-N-(4-hydroxy-bicyclo[2.2.2]octan-1-yl)-5-(4-((1R,5S)-3-(tetrahydro-2H-pyran-4-yl)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)nicotinamide or a pharmaceutically acceptable salt thereof, to a compound that is a pharmaceutically acceptable salt of 2-amino-N-(4-hydroxy-bicyclo[2.2.2]octan-1-yl)-5-(4-((1R,5S)-3-(tetrahydro-2H-pyran-4-yl)-3-azabicyclo[3.1.0]hexane-1-yl)phenyl)nicotinamide, where the pharmaceutically acceptable salt is selected from the group consisting of fumarate, phosphate, lactate, tartrate and malic acid salt and to a compound that is an anhydrate of 2-amino-N-(4-hydroxy-bicyclo[2.2.2]octan-1-yl)-5-(4-((1R,5S)-3-(tetrahydro-2H-pyran-4-yl)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)nicotinamide or a pharmaceutically acceptable salt thereof. The invention also relates to a pharmaceutical composition for inhibiting activin receptor-like kinase 2 (ALK-2) receptor activity in a subject, comprising a therapeutically effective amount of a compound of the invention and one or more pharmaceutically acceptable vehicles. The compounds of the invention are used in a method of inhibiting activin receptor-like kinase 2 (ALK-2) receptor activity in a subject, and of treating a disorder or disease selected from heterotopic ossification or progressive fibrodysplasia ossificans.

EFFECT: compounds being ALK-2 inhibitors.

22 cl, 7 tbl, 19 dwg, 94 ex

Description

BACKGROUND OF THE INVENTION

ALK–2, also known as activin A receptor type I (ACVR1) or serine–threonine protein kinase receptor R1 (SKR1), is a protein kinase encoded by the ACVR1 gene in humans.

ALK–2 is a type I BMP receptor that is widely expressed. It contains an extracellular ligand binding domain and a regulated intracellular serine/threonine kinase domain, both of which are required for signal transduction.

Bone morphogenetic proteins (BMPs) are multifunctional growth factors that are members of the transforming growth factor β (TGFβ) superfamily. BMP signaling plays an important role in the development of the heart, nervous system and cartilage, as well as bone formation in the postnatal period. BMPs ectopically induce endochondral bone formation and play a critical role in skeletal and joint morphogenesis (Urist, Science 110:893–899 (1965); Olsen et al. , Annu. Rev. Cell Dev. Biol. 16:191–220 (2000 ); Kronenberg, Nature 423:332–336 (2003); Thomas et al. , Nat. Genet. 12:315–317 (1996); Thomas et al. , Nat. Genet. 17:58–64 (1997); Polinkowsky et al. , Nat. Genet. 17:18–19 (1997); Storm et al ., Nature 368:639–643 (1994); and Wozney, Prog. Growth Factor Res. 1:267–280 (1989) ).

BMP signaling is regulated at multiple levels, including through extracellular antagonists such as noggin (Massague, Nat. Rev. MoI. Cell. Biol. 1:169–178 (2000)). It has been hypothesized that untimely or unwanted activation of signal transduction pathways essential for normal development may promote pathological processes such as spondyloarthropathies. The effects of BMP signaling on the initiation and progression of arthritis through genetic transfer of noggin have also been described (Lories et al. , J. Clin. Invest., 115, 1571–1579 (2005)). The physiological roles of BMP and BMP receptor signaling in normal bone formation, including skeletal and limb development, have been studied and reviewed in Zhao, Genetics 35:43–56 (2003).

Experiments with BMP antagonists demonstrate that regulation of BMP signaling proteins is a major factor in in vivo bone formation (Devlin et al ., Endocrinology 144:1972–1978 (2003) and Wu et al ., J. Clin. Invest., 112: 924 (2003)).

Fibrodysplasia ossificans progressiva (FOP) is a rare and disabling genetic disorder characterized by anatomical patterns of congenital big toe malformations and progressive heterotopic endochondral ossification. Ectopic expression of BMP4 has been identified in patients with FOP (Gannon et al., Hum. Pathol. 28:339–343 (1997) and Xu et al. , Clin. Genet. 58:291–298 (2000)). It has been shown that patients with FOP have activating mutations in ALK-2 (Shore et al ., Nat. Genet., 38(5):525–7 (2006)).

Excessive BMP signaling has been found to lead to a number of conditions described above. WO2008033408 and WO2009114180 describe inhibitors of the BMP signaling pathway. However, there is still an ongoing need to identify alternative pathways by which BMP signaling can be regulated.

This need may be met by the development of selective ALK-2 inhibitors.

Specific antibodies to ALK-2 are described, for example, in WO1994011502 and WO2008030611. Osteogenic proteins that bind to ALK-2 are described in WO2012023113 and WO2012077031.

WO2007123896 describes a method for treating pathology associated with heterotopic ossification by administering siRNA specific for a nucleic acid encoding mutant ALK-2.

WO2014160203 and WO2014138088 describe inhibitors of the BMP pathway. WO 2015152183 describes ALK-2 inhibitors for the treatment of FOP. ALK-2 inhibitors are also described in WO2014151871.

Brief description of the invention

There is a continuing need to develop new ALK–2 inhibitors that are good drug candidates. Such candidates may find applications inter alia in the treatment of fibrodysplasia ossificans progressiva (FOP) or non-hereditary heterotopic ossification (HO).

The present invention provides compounds, pharmaceutically acceptable salts thereof, pharmaceutical compositions containing them, wherein the compounds are ALK-2 inhibitors. The present invention further provides methods for treating, preventing, or reducing the severity of fibrodysplasia ossificans progressiva (FOP) or non-hereditary heterotopic ossification (HO), comprising administering to a subject in need thereof an effective amount of an ALK-2 inhibitor.

Various embodiments of the present invention are described herein.

In certain aspects, a compound of formula (I) is provided herein in free form or in pharmaceutically acceptable salt form

In another embodiment, the present invention provides a pharmaceutical composition containing a therapeutically effective amount of a compound of formula (I) as defined herein, in free form or in the form of a pharmaceutically acceptable salt or subformulas (Ia), (II), ( IIa), (IIb) and one or more pharmaceutically acceptable carriers.

In an additional aspect, the present invention relates to a method of inhibiting the activity of the ALK-2 receptor in a subject, wherein the method comprises administering to the subject a therapeutically effective amount of a compound of formula (I) as defined herein, in free form or in the form of a pharmaceutically acceptable salt or subformulas (Ia), (II), (IIa), (IIb) thereof.

In yet another aspect, the present invention provides a method of treating a disorder or disease selected from heterotopic ossification or fibrodysplasia ossificans progressiva, comprising administering to a subject a therapeutically effective amount of a compound of formula (I) as defined herein, in free form or in the form of a pharmaceutically acceptable salt or its subformulas (Ia), (II), (IIa), (IIb).

The compounds of the present invention exhibit acceptable pharmacokinetic properties, are non-toxic and exhibit few side effects. In particular, the compounds of the present invention are selective inhibitors of ALK-2 over other receptors. In addition, the ideal drug candidate will be in a form that is stable, non-hygroscopic and easy to formulate.

Figures

Figure 1 shows quantitative μCT analysis of heterotopic bone formed in the right hind limb of mice 6 weeks after adenovirus/CTX induction of FOP flare and shows the effect of treatment with Compound A for 6 weeks on HO formation. Mean +/- SEM. *: p < 0.05, **: p < 0.01, ***: p < 0.01 in comparison with treatment with carrier medium; one-way ANOVA using Dunnett's post hoc test.

Onfigure 2shows quantitative analysis using μCT of heterotopic bone formed in the right hind limb of mice 8 weeks after induction of FOP flare with adenovirus/CTX, showing the effect of treatment with Compound A for 6 weeks on HO formation. Mean +/- SEM. *: p < 0.05, **: p < 0.01, ***: p < 0.01 in comparison with treatment with carrier medium; Student's t-test.

Figure 3 shows a powder X-ray diffraction pattern of crystalline free form crystals of the H _A modification of compound A.

Figure 4 shows a differential scanning calorimetry diagram of crystals of the crystalline free form modification H _A of compound A.

Figure 5 shows a diagram of thermogravimetric analysis of the crystalline free form of the H _A modification of compound A.

Figure 6 shows a Fourier transform infrared spectroscopy diagram of the crystalline free form of the H _A modification of compound A.

Figure 7 shows a powder X-ray diffraction pattern of the crystalline free form of modification A of compound A.

Figure 8 shows a differential scanning calorimetry diagram of the crystalline free form of modification A of compound A.

Figure 9 shows a diagram of thermogravimetric analysis of the crystalline free form of modification A of compound A.

Figure 10 shows a Fourier transform infrared spectroscopy diagram of the crystalline free form of modification A of compound A.

Figure 11 shows a powder X-ray diffraction pattern of the crystalline fumaric acid salt of the H _A modification of compound A.

Figure 12 shows a differential scanning calorimetry diagram of a crystalline fumaric acid salt of the H _A modification of compound A.

Figure 13 shows a diagram of thermogravimetric analysis of the crystalline fumaric acid salt of the H _A modification of compound A.

Figure 14 shows the Fourier spectroscopy diagram in the infrared region of the crystalline fumaric acid salt of the H _A modification of compound A.

Figure 15 shows a diagram of the dynamic vapor sorption of crystalline fumaric acid salt modification H _A of compound A at 25 degrees C, method at relative humidity (RH) 50%–0%–90%.

Figure 16 shows a diagram of the dynamic vapor sorption of crystalline fumaric acid salt modification H _A of compound A at 40 degrees C, method at relative humidity (RH) 50%–0%–90%.

Figure 17 shows a powder X-ray diffraction pattern of a crystalline phosphoric acid salt of Compound A.

Figure 18 shows a powder X-ray diffraction pattern of the crystalline free form anhydride (Example 89 C).

Figure 19 shows the X-ray powder diffraction pattern of the crystalline free form trihydrate (Example 89 D)).

Detailed Description of the Invention

In particular, the present invention relates to a compound of formula (I) in free form or in the form of an acceptable salt,

Where

L represents the bond, (CH ₂ ) _n , –CH(CH ₃ )–, –O–(CH ₂ ) _n –, –C(O)–, –C(O)–NH–(CH ₂ ) _n – ;

n equals 1, 2, 3;

R ₁ is selected from C ₃ -C ₇ cycloalkyl, optionally substituted once or more than once with a substituent independently selected from hydroxyl, halogen, C ₁ -C ₃ alkyl; bridged C ₅ -C ₁₀ cycloalkyl, optionally substituted once or more than once with a substituent independently selected from hydroxyl, hydroxyC ₁ -C ₃ alkyl;

R ₂ and R ₃ are independently selected from H, halogen, C ₁ –C ₃ alkyl;

R ₄ is an N-containing heterocyclic non-aromatic ring, optionally containing one or more additional heteroatoms selected from N, O or S, wherein said ring is optionally substituted once or more than once with R ₇ ;

R ₇ is independently selected from C ₁ –C ₃ alkyl, hydroxyC ₁ –C ₃ alkyl, halogen _{C 1} –C ₃ alkyl, C ₁ –C ₃ alkoxyC ₁ –C ₃ alkyl, halogenC ₁ –C ₃ alkoxyC ₁ –C ₃ alkyl, C ₂ –C ₄ alkyl, cyanoC ₁ –C ₃ alkyl, (CH ₂ ) _m –R ₈ ;

m is 0, 1, 2 or 3;

R ₈ selected from:

– a 4-, 5- or 6-membered saturated or unsaturated non-aromatic heterocyclic ring containing one or more heteroatoms selected from N, O or S, wherein said ring is optionally substituted once or more than once with a substituent independently selected from oxo , SO ₂ C ₁ –C ₃ alkyl, halogen _{C 1} –C ₃ alkyl; or

– C ₃ –C ₆ cycloalkyl, optionally substituted once or more than once with halogen.

Unless otherwise indicated, the term "compounds of the present invention" or "compounds of the present invention" refers to the compounds of formula (I), (Ia), (II), (IIa), (IIb) and their salts, as well as all stereoisomers (including diastereoisomers and enantiomers), rotamers, tautomers and isotope-labeled compounds (including deuterium substitutions), as well as originally formed fragments.

As used herein, the term “C _1– C ₃ alkyl” refers to a linear or branched hydrocarbon chain radical consisting only of carbon and hydrogen atoms, containing no unsaturated bonds, having from one to three carbon atoms, and which is attached to the remainder molecules through a single bond. Examples of C1 _{– C3} alkyl include methyl, ethyl, n -propyl, 1-methylethyl ( isopropyl ).

As used herein, the term “hydroxyC _1– C ₃ alkyl” refers to a radical of the formula –R _a –OH, where R _a is C _1– C ₃ alkyl as defined above.

As used herein, the term “C _3– C ₇ cycloalkyl” refers to saturated monocyclic hydrocarbon groups of 3 to 7 carbon atoms. Examples of C3 _{- C7} cycloalkyl include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl.

As used herein, the term “C _{1 –} C ₃ alkoxy” refers to a radical of the formula –OR _a where R _a is a C _{1 –} C ₃ alkyl radical as generally defined above. Examples of C _{1 -} C ₃ alkoxy include, but are not limited to, methoxy, ethoxy, propoxy, isopropoxy, butoxy, isobutoxy, pentoxy and hexoxy.

As used herein, the term “C _1– C ₃ alkoxyC _1– C ₃ alkyl” refers to a radical of the formula –R _b –O–R _a , where R _a is a C _1– C ₃ alkyl radical and R _b is C _1– C ₃ alkyl radical as defined above. The oxygen atom can be bonded to any carbon atom in any alkyl radical. Examples of C _1– C ₃ alkoxyC _1– C ₃ alkyl include, for example, methoxymethyl, methoxyethyl, ethoxyethyl.

"Halogen" or "halogeno" refers to bromine, chlorine, fluorine or iodine.

As used herein, the term “haloC _1– C ₃ alkyl” or “haloC _1– C ₃ alkyl” refers to a C _1– C ₃ alkyl radical as defined above, substituted by one or more halogen radicals as defined above. Examples of halogenC1 _{- C3alkyl} include, for example, trifluoromethyl, difluoromethyl, fluoromethyl, trichloromethyl, 2,2,2-trifluoroethyl, 1-fluoromethyl-2-fluoroethyl, 3-bromo-2-fluoropropyl and 1-bromomethyl-2-bromoethyl .

As used herein, the term “haloC ₁ –C ₃ alkoxyC ₁ –C ₃ alkyl” refers to a radical of the formula –R _b –O–R _a where R _a is a C _1– C ₃ alkyl radical and R _b is a halogen C _1– C ₃ alkyl radical as defined above.

As used herein, the term "N-containing heterocyclic non-aromatic ring optionally containing one or more additional heteroatoms selected from N, O or S" refers to R₄ refers to a saturated or unsaturated non-aromatic ring or ring system which is a 4-, 5-, 6- or 7-membered monocyclic ring containing 1, 2 or 3 heteroatoms selected from O, S and N, where at least one a heteroatom is an N, 6-, 7-, 8-, 9-, 10-, 11- or 12-membered bicyclic ring system containing 1, 2, 3, 4 or 5 heteroatoms selected from O, S and N, where at least one heteroatom is N, or a 10-, 11-, 12-, 13-, 14- or 15-membered tricyclic ring system containing 1, 2, 3, 4, 5, 6 or 7 heteroatoms selected of O, S and N, wherein at least one heteroatom is N, wherein N and S may also optionally be oxidized to different oxidation states. An N-containing heterocyclic non-aromatic ring can be attached via a heteroatom or carbon atom. The N-containing heterocyclic non-aromatic ring may include fused or bridged rings, as well as spirocyclic rings. In a preferred embodiment, the "N-containing heterocyclic non-aromatic ring, optionally containing one or more additional heteroatoms selected from N, O or S" is an N-containing 5-membered saturated monocyclic ring, an N-containing 6-membered saturated monocyclic or bicyclic ring, N-containing a 7-membered saturated spirocyclic ring. Examples of N-containing heterocyclic non-aromatic ring include morpholine, piperazine, piperidine, imidazolidine, imidazoline, pyrroline, pyrrolidine, thiomorpholine, 3-azabicyclo[3.1.0]hexane.

As used herein, the term "N-containing 5-membered heterocyclic non-aromatic ring, optionally containing one or more additional heteroatoms selected from N, O or S" refers to R₄ includes, as examples, pyrrolidine, 2,3-dihydropyrrole, oxazolidine, imidazolidine. In a preferred embodiment, it refers to pyrrolidine. The ring is attached to the rest of the molecule via a carbon atom in the ring.

As used herein, the term "N-containing 6-membered heterocyclic non-aromatic monocyclic or bicyclic ring, optionally containing one or more additional heteroatoms selected from N, O or S" refers to R₄ includes, as examples, morpholine, thiomorpholine, piperidine, piperazine, 3-azabicyclo[3.1.0]hexane. In a preferred embodiment, it refers to 3-azabicyclo[3.1.0]hexane. The ring is attached to the rest of the molecule via a carbon atom in the ring.

As used herein, the term "5-membered ring optionally containing one additional heteroatom selected from N, O or S" refers to embodiments wherein R ₅ and R ₆ together with the N atom to which they are attached form said ring includes, as examples, pyrrolidine, 2,3-dihydropyrrole, oxazolidine, imidazolidine. In a preferred embodiment, it refers to pyrrolidine.

As used herein, the term "6-membered ring optionally containing one additional heteroatom selected from N, O or S" refers to embodiments wherein R ₅ and R ₆ together with the N atom to which they are attached form said ring includes, by way of example, piperidinyl, morpholinyl, piperazinyl.

As used herein, the term "7-membered spirocycle, optionally containing one additional heteroatom selected from N, O or S" refers to embodiments wherein _R5 and _R6 , together with the N atom to which they are attached, form said ring , includes, as examples, 2-azaspiro[3.3]heptane, 2-oxa-6-azaspiro[3.3]heptane.

As used herein, the term “bridged C ₅ –C ₁₀ cycloalkyl” refers to a saturated bicyclic or tricyclic ring system containing at least one bridge. Examples of bridged _C5 – _C10 cycloalkyl include, for example, bicyclo[1.1.1]pentane, bicyclo[2.2.2]octane, bicyclo[2.2.1]heptane, tricyclo[ ^3.3.1.13,7 ]decane.

As used herein, the term “C _2– C ₄ alkynyl” refers to a group that is a radical with a linear chain consisting of only carbon and hydrogen atoms, containing at least one triple bond, having from two to four carbon atoms, and which attached to the rest of the molecule via a single bond. Examples of C _2– C ₄ alkynyl include, for example, ethynyl, prop-1-ynyl, but-1-ynyl.

As used herein, the term “cyanoC ₁ –C ₃ alkyl” refers to a radical of the formula –R _a –CN, where R _a represents C _{1 –} C ₃ alkyl as defined above.

As used herein, the term "4-, 5-, or 6-membered saturated or unsaturated non-aromatic heterocyclic ring containing one or more heteroatoms selected from N, O or S" includes, as examples, tetrahydropyran, morpholine, piperidine, oxetane . In a preferred embodiment, it is tetrahydropyran.

As used herein, the term “ALK–2” refers to activin A type I receptor (ACVRI), also known as ACVRLK2; SKR1; ACVR1A; activin receptor type I; activin receptor-like kinase 2; serine/threonine protein kinase receptor R1; type I receptor of the TGF-B superfamily; ACTRI; TSRI; activin A receptor type II kinase 2; activin receptor type 1; hydroxyalkylprotein kinase; ACTR-I; TSR-I.

Various embodiments of the present invention are described herein. It will be appreciated that the features described in each embodiment may be combined with other described features to form additional embodiments of the present invention.

Embodiment 1. The compound of formula (I) in free form or in the form of a pharmaceutically acceptable salt,

Where

L represents the bond, (CH ₂ ) _n , –CH(CH ₃ )–, –O–(CH ₂ ) _n –, –C(O)–, –C(O)–NH–(CH ₂ ) _n – ;

n equals 1, 2, 3;

R ₁ is selected from C ₃ -C ₇ cycloalkyl, optionally substituted once or more than once with a substituent independently selected from hydroxyl, halogen, C ₁ -C ₃ alkyl; bridged C ₅ -C ₁₀ cycloalkyl, optionally substituted once or more than once with a substituent independently selected from hydroxyl, hydroxyC ₁ -C ₃ alkyl;

R ₂ and R ₃ are independently selected from H, halogen, C ₁ –C ₃ alkyl;

R ₄ is an N-containing heterocyclic non-aromatic ring, optionally containing one or more additional heteroatoms selected from N, O or S, wherein said ring is optionally substituted once or more than once with R ₇ ;

R ₇ is independently selected from C ₁ –C ₃ alkyl, hydroxyC ₁ –C ₃ alkyl, halogen _{C 1} –C ₃ alkyl, C ₁ –C ₃ alkoxyC ₁ –C ₃ alkyl, halogenC ₁ –C ₃ alkoxyC ₁ –C ₃ alkyl, C ₂ –C ₄ alkyl, cyanoC ₁ –C ₃ alkyl, (CH ₂ ) _m –R ₈ ;

m is 0, 1, 2 or 3;

R ₈ selected from:

– a 4-, 5- or 6-membered saturated or unsaturated non-aromatic heterocyclic ring containing one or more heteroatoms selected from N, O or S, wherein said ring is optionally substituted once or more than once with a substituent independently selected from oxo , SO ₂ C ₁ –C ₃ alkyl, halogen _{C 1} –C ₃ alkyl; or

– C ₃ –C ₆ cycloalkyl, optionally substituted once or more than once with halogen.

Embodiment 2. The compound according to Embodiment 1 of formula (Ia) in free form or in the form of a pharmaceutically acceptable salt,

Embodiment 3. The compound of any one of Embodiments 1 or 2 in free form or in the form of a pharmaceutically acceptable salt, wherein R ₄ is NR ₅ R ₆ wherein R ₅ and R ₆ together with the N atom to which they are attached form A 5-membered ring, optionally containing one additional heteroatom selected from N, O or S, optionally substituted once or more than once with R ₇ .

Embodiment 4. The compound of any one of Embodiments 1 or 2 in free form or in the form of a pharmaceutically acceptable salt, wherein R ₄ is NR ₅ R ₆ wherein R ₅ and R ₆ together with the N atom to which they are attached form A 6-membered ring, optionally containing one additional heteroatom selected from N, O or S, optionally substituted once or more than once with R ₇ .

Embodiment 5. The compound of any one of Embodiments 1 or 2 in free form or in the form of a pharmaceutically acceptable salt, wherein R ₄ is NR ₅ R ₆ wherein R ₅ and R ₆ together with the N atom to which they are attached form A 7-membered spirocycle, optionally containing one additional heteroatom selected from N, O or S, optionally substituted once or more than once with R ₇ .

Embodiment 6. The compound of any one of Embodiments 1 or 2 in free form or in pharmaceutically acceptable salt form, wherein R₄ is an N-containing 5-membered heterocyclic non-aromatic ring, optionally containing one or more additional heteroatoms selected from N, O or S, wherein said ring is optionally substituted once or more than once R₇ and where said ring is attached to the rest of the molecule via a carbon atom in the ring.

Embodiment 7. The compound of any one of Embodiments 1 or 2 in free form or in pharmaceutically acceptable salt form, wherein R₄ is an N-containing 6-membered heterocyclic non-aromatic monocyclic or bicyclic ring, optionally containing one or more additional heteroatoms selected from N, O or S, wherein said ring is optionally substituted once or more than once R₇ and where said ring is attached to the rest of the molecule via a carbon atom in the ring.

Embodiment 8. The compound according to any of embodiments 1, 2 or 7 of formula (II) in free form or in the form of a pharmaceutically acceptable salt,

where R ₇ is independently selected from C ₁ –C ₃ alkyl, hydroxyC ₁ –C ₃ alkyl, halogen _{C 1} –C ₃ alkyl, C ₁ –C ₃ alkoxyC _{1 –C 3} alkyl, halogenC ₁ –C ₃ alkoxyC ₁ –C ₃ alkyl , C ₂ –C ₄ alkynyl, cyanoC ₁ –C ₃ alkyl, (CH ₂ ) _m –R ₈ ;

m equals 0, 1, 2 or 3.

Embodiment 9. The compound according to embodiment 8 of formula (IIa) in free form or in the form of a pharmaceutically acceptable salt,

Embodiment 10. The compound according to embodiment 8 of formula (IIb) in free form or in the form of a pharmaceutically acceptable salt,

Embodiment 11. A compound according to any of the preceding embodiments in free form or in the form of a pharmaceutically acceptable salt, wherein:

R ₇ represents (CH ₂ ) _m –R ₈ ;

R ₈ is a 6-membered heterocyclic ring containing one or more heteroatoms selected from N, O or S, wherein said ring is optionally substituted once or more than once with a substituent independently selected from oxo, SO ₂ C ₁ –C ₃ alkyl, halogenC ₁ –C ₃ alkyl;

m equals 0, 1, 2 or 3.

Embodiment 12. The compound of Embodiment 11 in free or pharmaceutically acceptable salt form, wherein R ₈ is unsubstituted tetrahydropyran and m is 0.

Embodiment 13. The compound of Embodiment 11 in free form or in pharmaceutically acceptable salt form, wherein R ₈ is unsubstituted morpholine and m is 2 or 3.

Embodiment 14. A compound according to any of the preceding embodiments in free form or in the form of a pharmaceutically acceptable salt, wherein R ₁ is cyclohexyl, optionally substituted once or more than once with a substituent independently selected from hydroxyl, halogen, C ₁ –C ₃ alkyl.

Embodiment 15. A compound according to any of the preceding embodiments in free form or in the form of a pharmaceutically acceptable salt, wherein R ₁ is

.

Embodiment 16. A compound according to any of the preceding embodiments in free form or in the form of a pharmaceutically acceptable salt, wherein R ₁ is

.

Embodiment 17. The compound of any one of Embodiments 1 to 13 in free form or in pharmaceutically acceptable salt form, wherein R ₁ is bicyclo[2.2.2]octanyl, substituted in one case with hydroxyl.

Embodiment 18. The compound of Embodiment 17 in free form or in the form of a pharmaceutically acceptable salt, wherein R ₁ is .

Embodiment 19. The compound of embodiment, any of embodiments 1 to 13, in free form or in pharmaceutically acceptable salt form, wherein R ₁ is bicyclo[1.1.1]pentanyl substituted with hydroxymethyl.

Embodiment 20. The compound of Embodiment 19 in free form or in the form of a pharmaceutically acceptable salt, wherein R ₁ is

.

Embodiment 21. The compound of Embodiment 1 in free form or in pharmaceutically acceptable salt form, which is selected from

2-amino-N-(4-hydroxycyclohexyl)-5-(4-(3-(1-(methylsulfonyl)piperidin-4-yl)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)nicotinamide ;

2-amino-N-(4-hydroxycyclohexyl)-5-(4-(3-isopropyl-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)nicotinamide;

5-(4-(-1-(2-oxa-6-azaspiro[3.3]heptan-6-yl)ethyl)phenyl)-2-amino-N-(4-hydroxycyclohexyl)nicotinamide;

2-amino-N-(4-hydroxycyclohexyl)-5-(4-(3-(3-morpholinopropyl)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)nicotinamide;

2-amino-5-(4-(3-(1,3-difluoropropan-2-yl)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)-N-(4-hydroxycyclohexyl)nicotinamide;

2-amino-N-(4-hydroxybicyclo[2.2.2]octan-1-yl)-5-(4-(3-(2-methoxyethyl)-3-azabicyclo[3.1.0]hexan-1-yl) phenyl)nicotinamide;

2-amino-5-(2-fluoro-4-(4-isopropylpiperazin-1-yl)phenyl)-N-(4-hydroxycyclohexyl)nicotinamide;

2-amino-N-(4-hydroxycyclohexyl)-5-(4-(3-(2-morpholinoethyl)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)nicotinamide;

2-amino-5-(4-(3-(2-fluoroethyl)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)-N-(4-hydroxycyclohexyl)nicotinamide;

2-amino-N-(4-hydroxycyclohexyl)-5-(4-(1-isopropylpyrrolidin-3-yl)phenyl)nicotinamide;

2-amino-N-(4-hydroxy-4-methylcyclohexyl)-5-(4-(3-(tetrahydro-2H-pyran-4-yl)-3-azabicyclo[3.1.0]hexan-1-yl) phenyl)nicotinamide;

2-amino-N-(4-hydroxybicyclo[2.2.2]octan-1-yl)-5-(4-(3-(2-morpholinoethyl)-3-azabicyclo[3.1.0]hexan-1-yl) phenyl)nicotinamide;

2-amino-N-(3-(hydroxymethyl)bicyclo[1.1.1]pentan-1-yl)-5-(4-(3-isopropyl-3-azabicyclo[3.1.0]hexan-1-yl)phenyl )nicotinamide;

2-amino-N-(4-hydroxycyclohexyl)-5-(4-(3-(3,3,3-trifluoropropyl)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)nicotinamide;

2-amino-N-(4-hydroxycyclohexyl)-5-(4-(pyrrolidin-3-yl)phenyl)nicotinamide;

2-amino-N-(4-hydroxycyclohexyl)-5-(4-(3-(tetrahydro-2H-pyran-4-yl)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)nicotinamide;

2-amino-N-(4-hydroxycyclohexyl)-5-(4-(3-(2-methoxyethyl)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)nicotinamide;

2-amino-N-(3-fluoro-4-hydroxycyclohexyl)-5-(4-(3-(tetrahydro-2H-pyran-4-yl)-3-azabicyclo[3.1.0]hexan-1-yl) phenyl)nicotinamide;

5-(4-(2-azaspiro[3.3]heptan-2-ylmethyl)phenyl)-2-amino-N-(4-hydroxycyclohexyl)nicotinamide;

2-amino-N-(4-hydroxy-4-methylcyclohexyl)-5-(4-(3-isopropyl-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)nicotinamide;

2-amino-5-(4-(3-(4,4-difluorocyclohexyl)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)-N-(4-hydroxycyclohexyl)nicotinamide;

2-amino-N-(4-hydroxycyclohexyl)-5-(4-(3-(oxetan-3-yl)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)nicotinamide;

2-amino-N-(3-(hydroxymethyl)bicyclo[1.1.1]pentan-1-yl)-5-(4-(3-(tetrahydro-2H-pyran-4-yl)-3-azabicyclo[3.1 .0]hexan–1–yl)phenyl)nicotinamide;

2-amino-5-(2-fluoro-4-(piperazin-1-yl)phenyl)-N-(4-hydroxycyclohexyl)nicotinamide;

2-amino-N-(4-hydroxycyclohexyl)-5-(4-((2-methylpyrrolidin-1-yl)methyl)phenyl)nicotinamide;

2-amino-N-(4-hydroxycyclohexyl)-5-(4-(3-(tetrahydro-2H-pyran-4-yl)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)nicotinamide;

2-amino-N-(4-hydroxybicyclo[2.2.2]octan-1-yl)-5-(4-(3-(tetrahydro-2H-pyran-4-yl)-3-azabicyclo[3.1.0] hexan-1-yl)phenyl)nicotinamide;

2-amino-5-(4-(3-(4,4-difluorocyclohexyl)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)-N-(4-hydroxybicyclo[2.2.2]octane- 1-yl)nicotinamide;

2-amino-5-(4-(3-(but-2-in-1-yl)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)-N-(4-hydroxycyclohexyl)nicotinamide;

2-amino-N-(4-hydroxycyclohexyl)-5-(4-(3-(oxetan-3-ylmethyl)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)nicotinamide;

2-amino-N-(4-hydroxycyclohexyl)-5-(4-(3-(1-(2,2,2-trifluoroethyl)piperidin-4-yl)-3-azabicyclo[3.1.0]hexane-1 –yl)phenyl)nicotinamide;

2-amino-N-(4-hydroxycyclohexyl)-5-(4-(3-(2-(2,2,2-trifluoroethoxy)ethyl)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl )nicotinamide;

2-amino-N-cyclohexyl-5-(4-(3-(piperidin-1-yl)propoxy)phenyl)nicotinamide;

2-amino-5-(4-(3-(1,1-dioxidotetrahydro-2H-thiopyran-4-yl)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)-N-(4- hydroxycyclohexyl)nicotinamide;

2-amino-N-cyclohexyl-5-(4-(4-methylpiperazine-1-carbonyl)phenyl)nicotinamide;

2-amino-5-(3-fluoro-4-(2-methylpyrrolidin-1-yl)methyl)phenyl)-N-(4-hydroxy-4-methylcyclohexyl)nicotinamide;

2-amino-N-cyclohexyl-5-(4-morpholinophenyl)nicotinamide;

2-amino-5-(4-(3-(2,2-difluoropropyl)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)-N-(4-hydroxycyclohexyl)nicotinamide;

2-amino-5-(4-(3-(2-cyanoethyl)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)-N-(4-hydroxycyclohexyl)nicotinamide;

2-amino-N-cyclohexyl-5-(4-(3-morpholinopropoxy)phenyl)nicotinamide;

2-amino-N-(4-hydroxycyclohexyl)-5-(4-(3-(prop-2-yn-1-yl)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)nicotinamide;

5-(4-(1-(2-oxa-6-azaspiro[3.3]heptan-6-yl)ethyl)-2-chlorophenyl)-2-amino-N-(4-hydroxycyclohexyl)nicotinamide;

2-amino-5-(2-fluoro-4-((2-methylpyrrolidin-1-yl)methyl)phenyl)-N-(4-hydroxy-4-methylcyclohexyl)nicotinamide;

2-amino-5-(4-(3-(2-fluoroethyl)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)-N-(4-hydroxybicyclo[2.2.2]octane-1- yl)nicotinamide;

2-amino-5-(4-(3-(4,4-difluorocyclohexyl)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)-N-(4-hydroxy-4-methylcyclohexyl)nicotinamide;

5-(4-(1-(2-oxa-6-azaspiro[3.3]heptan-6-yl)ethyl)phenyl)-2-amino-N-(4-hydroxy-4-methylcyclohexyl)nicotinamide;

2-amino-N-cyclohexyl-5-(4-(piperidin-1-yl)phenyl)nicotinamide;

2-amino-N-(4-hydroxy-4-methylcyclohexyl)-5-(4-(3-(tetrahydro-2H-pyran-4-yl)-3-azabicyclo[3.1.0]hexan-1-yl) phenyl)nicotinamide;

2-amino-N-(4-hydroxybicyclo[2.2.2]octan-1-yl)-5-(4-(3-(oxetan-3-yl)-3-azabicyclo[3.1.0]hexane-1- yl)phenyl)nicotinamide;

2-amino-N-cyclohexyl-5-(4-((2-(4-methylpiperazin-1-yl)ethyl)carbamoyl)phenyl)nicotinamide;

2-amino-N-(4-hydroxycyclohexyl)-5-(4-(3-(2,2,2-trifluoroethyl)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)nicotinamide;

5-(4-(-1-(2-oxa-6-azaspiro[3.3]heptan-6-yl)ethyl)phenyl)-2-amino-N-(3-hydroxyadamantan-1-yl)nicotinamide;

2-amino-N-cyclohexyl-5-(4-(morpholine-4-carbonyl)phenyl)nicotinamide;

2-amino-N-(4-hydroxy-4-methylcyclohexyl)-5-(4-((2-methylpyrrolidin-1-yl)methyl)phenyl)nicotinamide;

2-amino-5-(2,3-difluoro-4-((2-methylpyrrolidin-1-yl)methyl)phenyl)-N-(4-hydroxy-4-methylcyclohexyl)nicotinamide;

2-amino-N-(4-hydroxycyclohexyl-4-d)-5-(4-(3-isopropyl-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)nicotinamide;

2-amino-N-(4-hydroxycyclohexyl)-5-(4-(4-methylpiperazine-1-carbonyl)phenyl)nicotinamide;

2-amino-5-(2-chloro-4-((2-methylpyrrolidin-1-yl)methyl)phenyl)-N-(4-hydroxy-4-methylcyclohexyl)nicotinamide;

2-amino-N-(3-(2-hydroxypropan-2-yl)bicyclo[1.1.1]pentan-1-yl)-5-(4-(3-(tetrahydro-2H-pyran-4-yl) –3-azabicyclo[3.1.0]hexane-1-yl)phenyl)nicotinamide;

2-amino-N-cyclohexyl-5-(3-(morpholine-4-carbonyl)phenyl)nicotinamide;

2-amino-5-(3-chloro-4-((2-methylpyrrolidin-1-yl)methyl)phenyl)-N-(4-hydroxy-4-methylcyclohexyl)nicotinamide;

5-(4-(1-(2-oxa-6-azaspiro[3.3]heptan-6-yl)ethyl)phenyl)-2-amino-N-(4-hydroxy-1-methylcyclohexyl)nicotinamide;

2-amino-N-(4-hydroxy-4-methylcyclohexyl)-5-(2-methyl-4-((2-methylpyrrolidin-1-yl)methyl)phenyl)nicotinamide and

2-amino-N-cyclohexyl-5-(3-(4-(2-hydroxyethyl)piperazine-1-carbonyl)phenyl)nicotinamide.

Embodiment 22. The compound of Embodiment 21 in free form or in pharmaceutically acceptable salt form, which is selected from

2-amino-N-((1r,4R)-4-hydroxycyclohexyl)-5-(4-((1R,5S)-3-(1-(methylsulfonyl)piperidin-4-yl)-3-azabicyclo[3.1 .0]hexan–1–yl)phenyl)nicotinamide;

2-amino-N-((1r,4R)-4-hydroxycyclohexyl)-5-(4-((1R,5S)-3-isopropyl-3-azabicyclo[3.1.0]hexan-1-yl)phenyl) nicotinamide;

5-(4-((R)-1-(2-oxa-6-azaspiro[3.3]heptan-6-yl)ethyl)phenyl)-2-amino-N-((1r,4R)-4-hydroxycyclohexyl )nicotinamide;

5-(4-((S)-1-(2-oxa-6-azaspiro[3.3]heptan-6-yl)ethyl)phenyl)-2-amino-N-((1r,4R)-4-hydroxycyclohexyl )nicotinamide;

2-amino-N-((1r,4S)-4-hydroxycyclohexyl)-5-(4-((1S,5R)-3-(3-morpholinopropyl)-3-azabicyclo[3.1.0]hexane-1- yl)phenyl)nicotinamide;

2-amino-5-(4-((1R,5S)-3-(1,3-difluoropropan-2-yl)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)-N-( (1r,4R)–4-hydroxycyclohexyl)nicotinamide;

2-amino-N-(4-hydroxybicyclo[2.2.2]octan-1-yl)-5-(4-((1S,5R)-3-(2-methoxyethyl)-3-azabicyclo[3.1.0] hexan-1-yl)phenyl)nicotinamide;

2-amino-5-(2-fluoro-4-(4-isopropylpiperazin-1-yl)phenyl)-N-((1r,4r)-4-hydroxycyclohexyl)nicotinamide;

2-amino-N-((1r,4S)-4-hydroxycyclohexyl)-5-(4-((1S,5R)-3-isopropyl-3-azabicyclo[3.1.0]hexan-1-yl)phenyl) nicotinamide;

2-amino-N-((1r,4R)-4-hydroxycyclohexyl)-5-(4-((1R,5S)-3-(2-morpholinoethyl)-3-azabicyclo[3.1.0]hexane-1- yl)phenyl)nicotinamide;

2-amino-5-(4-((1S,5R)-3-(2-fluoroethyl)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)-N-((1r,4S)- 4-hydroxycyclohexyl)nicotinamide;

2-amino-N-((1r,4r)-4-hydroxycyclohexyl)-5-(4-(1-isopropylpyrrolidin-3-yl)phenyl)nicotinamide;

2-amino-N-((1r,4S)-4-hydroxycyclohexyl)-5-(4-((1S,5R)-3-(2-morpholinoethyl)-3-azabicyclo[3.1.0]hexane-1- yl)phenyl)nicotinamide;

2-amino-N-((1r,4R)-4-hydroxy-4-methylcyclohexyl)-5-(4-((1R,5S)-3-(tetrahydro-2H-pyran-4-yl)-3- azabicyclo[3.1.0]hexan–1–yl)phenyl)nicotinamide;

2-amino-N-(4-hydroxybicyclo[2.2.2]octan-1-yl)-5-(4-((1R,5S)-3-(2-morpholinoethyl)-3-azabicyclo[3.1.0] hexan-1-yl)phenyl)nicotinamide;

2-amino-N-(3-(hydroxymethyl)bicyclo[1.1.1]pentan-1-yl)-5-(4-((1S,5R)-3-isopropyl-3-azabicyclo[3.1.0]hexane –1-yl)phenyl)nicotinamide;

2-amino-N-((1r,4S)-4-hydroxycyclohexyl)-5-(4-((1S,5R)-3-(3,3,3-trifluoropropyl)-3-azabicyclo[3.1.0] hexan-1-yl)phenyl)nicotinamide;

2-amino-N-((1r,4r)-4-hydroxycyclohexyl)-5-(4-(pyrrolidin-3-yl)phenyl)nicotinamide;

2-amino-N-((1r,4R)-4-hydroxycyclohexyl)-5-(4-((1R,5S)-3-(tetrahydro-2H-pyran-4-yl)-3-azabicyclo[3.1. 0]hexan–1–yl)phenyl)nicotinamide;

2-amino-N-((1r,4S)-4-hydroxycyclohexyl)-5-(4-((1S,5R)-3-(2-methoxyethyl)-3-azabicyclo[3.1.0]hexane-1- yl)phenyl)nicotinamide;

2-amino-N-((1R,3S,4R)-3-fluoro-4-hydroxycyclohexyl)-5-(4-((1R,5S)-3-(tetrahydro-2H-pyran-4-yl)- 3-azabicyclo[3.1.0]hexane-1-yl)phenyl)nicotinamide;

2-amino-N-((1s,4S)-4-hydroxycyclohexyl)-5-(4-((1R,5S)-3-(tetrahydro-2H-pyran-4-yl)-3-azabicyclo[3.1. 0]hexan–1–yl)phenyl)nicotinamide;

5-(4-(2-azaspiro[3.3]heptan-2-ylmethyl)phenyl)-2-amino-N-((1r,4r)-4-hydroxycyclohexyl)nicotinamide;

2-amino-N-((1r,4R)-4-hydroxy-4-methylcyclohexyl)-5-(4-((1R,5S)-3-isopropyl-3-azabicyclo[3.1.0]hexane-1- yl)phenyl)nicotinamide;

2-amino-5-(4-((1R,5S)-3-(4,4-difluorocyclohexyl)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)-N-((1r,4R )–4-hydroxycyclohexyl)nicotinamide;

2-amino-N-((1r,4S)-4-hydroxycyclohexyl)-5-(4-((1S,5R)-3-(oxetan-3-yl)-3-azabicyclo[3.1.0]hexane- 1-yl)phenyl)nicotinamide;

2-amino-5-(4-((1S,5R)-3-(4,4-difluorocyclohexyl)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)-N-((1r,4S )–4-hydroxycyclohexyl)nicotinamide;

2-amino-N-(3-(hydroxymethyl)bicyclo[1.1.1]pentan-1-yl)-5-(4-((1S,5R)-3-(tetrahydro-2H-pyran-4-yl) –3-azabicyclo[3.1.0]hexane-1-yl)phenyl)nicotinamide;

2-amino-5-(2-fluoro-4-(piperazin-1-yl)phenyl)-N-((1r,4r)-4-hydroxycyclohexyl)nicotinamide;

2-amino-N-(3-(hydroxymethyl)bicyclo[1.1.1]pentan-1-yl)-5-(4-((1R,5S)-3-(tetrahydro-2H-pyran-4-yl) –3-azabicyclo[3.1.0]hexane-1-yl)phenyl)nicotinamide;

2-amino-N-((1r,4R)-4-hydroxycyclohexyl)-5-(4-(((R)-2-methylpyrrolidin-1-yl)methyl)phenyl)nicotinamide;

2-amino-N-((1r,4S)-4-hydroxy-4-methylcyclohexyl)-5-(4-((1S,5R)-3-isopropyl-3-azabicyclo[3.1.0]hexane-1- yl)phenyl)nicotinamide;

2-amino-N-((1r,4S)-4-hydroxycyclohexyl)-5-(4-((1S,5R)-3-(tetrahydro-2H-pyran-4-yl)-3-azabicyclo[3.1. 0]hexan–1–yl)phenyl)nicotinamide;

2-amino-N-(4-hydroxybicyclo[2.2.2]octan-1-yl)-5-(4-((1R,5S)-3-(tetrahydro-2H-pyran-4-yl)-3- azabicyclo[3.1.0]hexan–1–yl)phenyl)nicotinamide;

2-amino-5-(4-((1S,5R)-3-(4,4-difluorocyclohexyl)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)-N-(4-hydroxybicyclo[ 2.2.2]octan–1–yl)nicotinamide;

2-amino-N-(3-(hydroxymethyl)bicyclo[1.1.1]pentan-1-yl)-5-(4-((1R,5S)-3-isopropyl-3-azabicyclo[3.1.0]hexane –1-yl)phenyl)nicotinamide;

2-amino-5-(4-((1S,5R)-3-(but-2-in-1-yl)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)-N-( (1r,4S)–4-hydroxycyclohexyl)nicotinamide;

2-amino-N-((1r,4S)-4-hydroxycyclohexyl)-5-(4-((1S,5R)-3-(oxetan-3-ylmethyl)-3-azabicyclo[3.1.0]hexane- 1-yl)phenyl)nicotinamide;

2-amino-N-((1r,4R)-4-hydroxycyclohexyl)-5-(4-((1R,5S)-3-(1-(2,2,2-trifluoroethyl)piperidin-4-yl) –3-azabicyclo[3.1.0]hexane-1-yl)phenyl)nicotinamide;

2-amino-N-(4-hydroxybicyclo[2.2.2]octan-1-yl)-5-(4-((1S,5R)-3-(2-morpholinoethyl)-3-azabicyclo[3.1.0] hexan-1-yl)phenyl)nicotinamide;

2-amino-N-((1r,4R)-4-hydroxycyclohexyl)-5-(4-((1R,5S)-3-(2-(2,2,2-trifluoroethoxy)ethyl)-3-azabicyclo [3.1.0]hexan–1–yl)phenyl)nicotinamide;

2-amino-N-(4-hydroxybicyclo[2.2.2]octan-1-yl)-5-(4-((1S,5R)-3-(tetrahydro-2H-pyran-4-yl)-3- azabicyclo[3.1.0]hexan–1–yl)phenyl)nicotinamide;

2-amino-5-(4-((1R,5S)-3-(1,1-dioxidotetrahydro-2H-thiopyran-4-yl)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl) –N–((1r,4R)–4–hydroxycyclohexyl)nicotinamide;

2-amino-5-(3-fluoro-4-(((R)-2-methylpyrrolidin-1-yl)methyl)phenyl)-N-((1r,4R)-4-hydroxy-4-methylcyclohexyl)nicotinamide ;

2-amino-N-((1r,4R)-4-hydroxycyclohexyl)-5-(4-((1R,5S)-3-(3,3,3-trifluoropropyl)-3-azabicyclo[3.1.0] hexan-1-yl)phenyl)nicotinamide;

2-amino-5-(4-((1R,5S)-3-(2-fluoroethyl)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)-N-((1r,4R)- 4-hydroxycyclohexyl)nicotinamide;

2-amino-5-(4-((1R,5S)-3-(2,2-difluoropropyl)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)-N-((1r,4R )–4-hydroxycyclohexyl)nicotinamide;

2-amino-5-(4-((1S,5R)-3-(2-cyanoethyl)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)-N-((1r,4S)- 4-hydroxycyclohexyl)nicotinamide;

2-amino-N-((1r,4S)-4-hydroxycyclohexyl)-5-(4-((1S,5R)-3-(prop-2-in-1-yl)-3-azabicyclo[3.1. 0]hexan–1–yl)phenyl)nicotinamide;

2-amino-N-((1r,4R)-4-hydroxycyclohexyl)-5-(4-((1R,5S)-3-(prop-2-in-1-yl)-3-azabicyclo[3.1. 0]hexan–1–yl)phenyl)nicotinamide;

5-(4-((R)-1-(2-oxa-6-azaspiro[3.3]heptan-6-yl)ethyl)-2-chlorophenyl)-2-amino-N-((1r,4R)- 4-hydroxycyclohexyl)nicotinamide;

2-amino-5-(2-fluoro-4-(((R)-2-methylpyrrolidin-1-yl)methyl)phenyl)-N-((1r,4R)-4-hydroxy-4-methylcyclohexyl)nicotinamide ;

2-amino-5-(4-((1R,5S)-3-(2-fluoroethyl)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)-N-(4-hydroxybicyclo[2.2. 2]octan–1–yl)nicotinamide;

2-amino-5-(4-((1S,5R)-3-(4,4-difluorocyclohexyl)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)-N-((1r,4S )-4-hydroxy-4-methylcyclohexyl)nicotinamide;

5-(4-((R)-1-(2-oxa-6-azaspiro[3.3]heptan-6-yl)ethyl)phenyl)-2-amino-N-((1r,4R)-4-hydroxy –4-methylcyclohexyl)nicotinamide;

2-amino-5-(4-((1R,5S)-3-(but-2-in-1-yl)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)-N-( (1r,4R)–4-hydroxycyclohexyl)nicotinamide;

2-amino-N-((1r,4S)-4-hydroxy-4-methylcyclohexyl)-5-(4-((1S,5R)-3-(tetrahydro-2H-pyran-4-yl)-3- azabicyclo[3.1.0]hexan–1–yl)phenyl)nicotinamide;

2-amino-N-(4-hydroxybicyclo[2.2.2]octan-1-yl)-5-(4-((1S,5R)-3-(oxetan-3-yl)-3-azabicyclo[3.1. 0]hexan–1–yl)phenyl)nicotinamide;

2-amino-N-(4-hydroxybicyclo[2.2.2]octan-1-yl)-5-(4-((1R,5S)-3-(oxetan-3-yl)-3-azabicyclo[3.1. 0]hexan–1–yl)phenyl)nicotinamide;

2-amino-N-cyclohexyl-5-(4-((2-(4-methylpiperazin-1-yl)ethyl)carbamoyl)phenyl)nicotinamide;

2-amino-5-(4-((1S,5R)-3-(1,3-difluoropropan-2-yl)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)-N-( (1r,4S)–4-hydroxycyclohexyl)nicotinamide;

2-amino-N-((1r,4S)-4-hydroxycyclohexyl)-5-(4-((1S,5R)-3-(2,2,2-trifluoroethyl)-3-azabicyclo[3.1.0] hexan-1-yl)phenyl)nicotinamide;

5-(4-((R)-1-(2-oxa-6-azaspiro[3.3]heptan-6-yl)ethyl)phenyl)-2-amino-N-((1R,3R)-3-hydroxyadamantane –1-yl)nicotinamide;

2-amino-N-((1r,4R)-4-hydroxycyclohexyl)-5-(4-((1R,5S)-3-(oxetan-3-ylmethyl)-3-azabicyclo[3.1.0]hexane- 1-yl)phenyl)nicotinamide;

2-amino-N-((1s,4R)-4-hydroxycyclohexyl)-5-(4-((1S,5R)-3-isopropyl-3-azabicyclo[3.1.0]hexan-1-yl)phenyl) nicotinamide;

2-amino-N-((1r,4R)-4-hydroxy-4-methylcyclohexyl)-5-(4-(((R)-2-methylpyrrolidin-1-yl)methyl)phenyl)nicotinamide;

2-amino-5-(4-((1S,5R)-3-(2-fluoroethyl)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)-N-(4-hydroxybicyclo[2.2. 2]octan–1–yl)nicotinamide;

2-amino-5-(2,3-difluoro-4-(((R)-2-methylpyrrolidin-1-yl)methyl)phenyl)-N-((1r,4R)-4-hydroxy-4-methylcyclohexyl )nicotinamide;

2-amino-N-((1r,4R)-4-hydroxycyclohexyl)-5-(4-((1R,5S)-3-(2-methoxyethyl)-3-azabicyclo[3.1.0]hexane-1- yl)phenyl)nicotinamide;

2-amino-5-(4-((1S,5R)-3-(2,2-difluoropropyl)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)-N-((1r,4S )–4-hydroxycyclohexyl)nicotinamide;

2-amino-N-((1r,4R)-4-hydroxycyclohexyl)-5-(4-((1R,5S)-3-(2,2,2-trifluoroethyl)-3-azabicyclo[3.1.0] hexan-1-yl)phenyl)nicotinamide;

2-amino-N-((1r,4S)-4-hydroxycyclohexyl-4-d)-5-(4-((1S,5R)-3-isopropyl-3-azabicyclo[3.1.0]hexane-1- yl)phenyl)nicotinamide;

2-amino-N-((1s,4s)-4-hydroxycyclohexyl)-5-(4-(4-methylpiperazine-1-carbonyl)phenyl)nicotinamide;

2-amino-5-(4-((1R,5S)-3-(4,4-difluorocyclohexyl)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)-N-(4-hydroxybicyclo[ 2.2.2]octan–1–yl)nicotinamide;

2-amino-5-(2-chloro-4-(((R)-2-methylpyrrolidin-1-yl)methyl)phenyl)-N-((1r,4R)-4-hydroxy-4-methylcyclohexyl)nicotinamide ;

2-amino-5-(4-((1R,5S)-3-(4,4-difluorocyclohexyl)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)-N-((1r,4R )-4-hydroxy-4-methylcyclohexyl)nicotinamide;

2-amino-N-(3-(2-hydroxypropan-2-yl)bicyclo[1.1.1]pentan-1-yl)-5-(4-((1R,5S)-3-(tetrahydro-2H- pyran-4-yl)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)nicotinamide;

2-amino-5-(3-chloro-4-(((R)-2-methylpyrrolidin-1-yl)methyl)phenyl)-N-((1r,4R)-4-hydroxy-4-methylcyclohexyl)nicotinamide ;

5-(4-((R)-1-(2-oxa-6-azaspiro[3.3]heptan-6-yl)ethyl)phenyl)-2-amino-N-((1r,4R)-4-hydroxy –1-methylcyclohexyl)nicotinamide and

2-amino-N-((1r,4R)-4-hydroxy-4-methylcyclohexyl)-5-(2-methyl-4-(((R)-2-methylpyrrolidin-1-yl)methyl)phenyl)nicotinamide .

Embodiment 23. A pharmaceutical composition comprising a therapeutically effective amount of a compound according to any of the preceding embodiments in free or pharmaceutically acceptable salt form and one or more pharmaceutically acceptable carriers.

Embodiment 24. A combination comprising a therapeutically effective amount of a compound according to any of embodiments 1-22 in free form or in pharmaceutically acceptable salt form and one or more therapeutically active agents.

Embodiment 25. The compound according to any one of embodiments 1 to 22 in free form or in the form of a pharmaceutically acceptable salt for use as a drug.

Embodiment 26. The compound of any one of Embodiments 1-22 in free form or in pharmaceutically acceptable salt form for use in the treatment of a disorder or disease selected from heterotopic ossification or fibrodysplasia ossificans progressiva.

Embodiment 27. 2-Amino-N-(4-hydroxybicyclo[2.2.2]octan-1-yl)-5-(4-((1R,5S)-3-(tetrahydro-2H-pyran-4-yl )-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)nicotinamide in free form, in particular in the free form of the _HA modification, in particular (i) characterized by 2-theta values in the XRPD reflection of the first 2, the first 3, the first 4, the first 5, the first 6, the first 8, in particular the first ten, or in particular all the 2-theta values given in Table A below; or (ii) showing the DSC diagram shown in FIG. 4; or (iii) characterized by a TGA diagram shown in FIG. 5; or (iv) characterized by the FT-IR diagram shown in FIG. 6; or characterized by two, or three, or in particular all of the properties (i)–(iv) just mentioned; or, in particular, characterized by the XRPD diagram shown in FIG. 3.

Embodiment 28. 2-Amino-N-(4-hydroxybicyclo[2.2.2]octan-1-yl)-5-(4-((1R,5S)-3-(tetrahydro-2H-pyran-4-yl )-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)nicotinamide in free form, in particular in the form of modification A, in particular, (i) characterized by 2-theta values in the XRPD reflection of the first 2, first 3 , the first 4, the first 5, the first 6, the first 8, particularly the first ten, or all the 2-theta values given in Table B below; or (ii) showing the DSC diagram shown in FIG. 8; or (iii) characterized by a TGA diagram shown in FIG. 9; or (iv) characterized by the FT-IR diagram shown in FIG. 10; or characterized by two, or three, or in particular all of the properties (i)–(iv) just mentioned; or, in particular, characterized by the XRPD diagram shown in FIG. 7.

Embodiment 29. 2-Amino-N-(4-hydroxybicyclo[2.2.2]octan-1-yl)-5-(4-((1R,5S)-3-(tetrahydro-2H-pyran-4-yl )-3-azabicyclo[3.1.0]hexane-1-yl)phenyl)nicotinamide as the free form anhydride, particularly characterized by the XRPD diagram shown in FIG. 18.

Embodiment 30. 2-Amino-N-(4-hydroxybicyclo[2.2.2]octan-1-yl)-5-(4-((1R,5S)-3-(tetrahydro-2H-pyran-4-yl )-3-azabicyclo[3.1.0]hexane-1-yl)phenyl)nicotinamide as the free form trihydrate, particularly characterized by the XRPD diagram shown in Figure 19.

Embodiment 31. 2-Amino-N-(4-hydroxybicyclo[2.2.2]octan-1-yl)-5-(4-((1R,5S)-3-(tetrahydro-2H-pyran-4-yl )-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)nicotinamide in the form of a fumaric acid salt, in particular characterized by a 1:1 molar ratio of compound A and fumaric acid, in particular in the form of a fumaric acid salt acid modification _HA , in particular, (i) characterized by 2-theta values in the XRPD reflection of the first 2, first 3, first 4, first 5, first 6, first 8, in particular the first ten, or, in particular, all values 2–theta, shown in Table C below; or (ii) showing the DSC diagram shown in FIG. 12; or (iii) characterized by a TGA diagram shown in FIG. 13; or (iv) characterized by the FT-IR diagram shown in FIG. 14; or (v) characterized by the DVS diagram at 25°C shown in FIG. 15; or (vi) characterized by the DVS diagram at 40°C shown in FIG. 16; or characterized by two, or three, or four, or five, or in particular all of the properties (i)–(iv) just mentioned; or, in particular, characterized by the XRPD diagram shown in FIG. eleven.

Embodiment 32. 2-Amino-N-(4-hydroxybicyclo[2.2.2]octan-1-yl)-5-(4-((1R,5S)-3-(tetrahydro-2H-pyran-4-yl )-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)nicotinamide as fumaric acid amorphous salt form variant 1, in particular characterized by a 1:1 molar ratio of compound A and fumaric acid, in particular demonstrating glass transition at approximately 143°C when analyzed by modulated DSC at a heating rate of 1 K/min, temperature peak of 1 K, period of 60 seconds.

Embodiment 33. 2-Amino-N-(4-hydroxybicyclo[2.2.2]octan-1-yl)-5-(4-((1R,5S)-3-(tetrahydro-2H-pyran-4-yl )-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)nicotinamide as variant 2 of the amorphous fumaric acid salt form, in particular characterized by a 1:1 molar ratio of compound A and fumaric acid, in particular, showing a glass transition at approximately 78°C when analyzed by DSC at a heating rate of 10 K/min.

Embodiment 34. 2-Amino-N-(4-hydroxybicyclo[2.2.2]octan-1-yl)-5-(4-((1R,5S)-3-(tetrahydro-2H-pyran-4-yl )-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)nicotinamide in the form of a phosphoric acid salt, in particular characterized by a 1:1.5 molar ratio of compound A and orthophosphoric acid, in particular (i ) characterized by the 2-theta values in the XRPD reflection of the first 2, the first 3, the first 4, the first 5, the first 6, the first 8, in particular the first ten, or in particular all of the 2-theta values given in Table D below; or, in particular, characterized by the XRPD diagram shown in FIG. 17.

Embodiment 35: The compound is in the form of any one of embodiments 27 to 34 for use as a drug.

Embodiment 36. A compound in the form of any one of embodiments 27-34 for use in treating a disorder or disease selected from heterotopic ossification or fibrodysplasia ossificans progressiva.

Where the value 2-theta (or "angle" in the tables below) is given in this invention, it means a relative value of ±0.2, or more specifically the value as such.

Where the term "modification" is used, it refers to amorphous forms or, in particular, to crystalline polymorphs (i.e. forms of crystalline structures), also including pseudo-polymorphs such as solvates or, in particular, hydrates of free forms or salts, respectively .

Depending on the choice of starting materials and procedures, compounds can be present in the form of pure optical isomers or as mixtures of isomers, such as racemates and mixtures of diastereoisomers, depending on the number of asymmetric carbon atoms. The present invention is intended to include all such possible isomers, including racemic mixtures, mixtures of diastereoisomers, and optically pure forms. Optically active ( R )- and ( S )-isomers can be prepared using chiral synthons or chiral reagents or isolated using conventional techniques. If the compound contains a double bond, the substituent may have an E- or Z-configuration. If the compound contains a disubstituted cycloalkyl, the cycloalkyl substituent may have a cis or trans configuration. It is also intended to include all tautomeric forms.

As used herein, the terms “salt” or “salts” refer to the acid addition or base addition salt of a compound of the present invention. "Salts" include, in particular, "pharmaceutically acceptable salts". The term "pharmaceutically acceptable salts" refers to salts that retain the biological effectiveness and properties of the compounds of this invention and that are generally not biologically or otherwise undesirable. In many cases, the compounds of the present invention are capable of forming acid salts due to the presence of a basic aminopyridine moiety. Where "free form" is stated, this refers to the form without additional acids and/or bases, i.e. the compound itself (which, however, can form internal salts if the compound contains basic and acidic groups).

Pharmaceutically acceptable acid addition salts can be formed using inorganic acids and organic acids.

Inorganic acids from which salts can be prepared include, for example, hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid and the like.

Organic acids from which salts can be prepared include, for example, acetic acid, propionic acid, glycolic acid, oxalic acid, maleic acid, malonic acid, succinic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, toluenesulfonic acid, sulfosalicylic acid and the like.

In another aspect, the present invention provides compounds of formula (I) in the form of a salt of acetate, ascorbate, adipate, aspartate, benzoate, besylate, bromide/hydrobromide, bicarbonate/carbonate, bisulfate/sulfate, camphorsulfonic acid salt, capric acid salt, chloride /hydrochloride, chlortheophyllonate, citrate, ethanesulfonate, fumarate, gluceptate, gluconate, glucuronate, glutamate, glutarate, glycolate, hippurate, hydroiodide/iodide, isethionate, lactate, lactobionate, lauryl sulfate, malic acid salt, maleic acid salt, malonate, mandelic acid salt , mesylate, methyl sulfate, mucinic acid salt, naphthoic acid salt, napsylate, nicotinate, nitrate, octadecanoate, oleate, oxalate, palmitate, pamoate, phosphate/hydrogen phosphate/dihydrogen phosphate, polygalacturonate, propionate, sebacate, stearate, succinate, sulfosalicylate, sulfate, tartrate , tosylate triphenatate, trifluoroacetate or xinafoate.

Any formula given herein also implies the presence of unlabeled forms as well as isotopically labeled forms of the compounds. Isotopically labeled compounds have the structures depicted by the formulas given herein, except that one or more atoms are replaced by an atom having the selected atomic mass or mass number. Examples of isotopes that may be included in the compounds of the present invention include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, fluorine and chlorine, such as ² H, ³ H, ¹¹ C, ¹³ C, ¹⁴ C, ¹⁵ N, ¹⁸ F ³¹ P, ³² P, ³⁵ S, ³⁶ Cl, ¹²³ I, ¹²⁴ I, ¹²⁵ I, respectively. The present invention includes various isotopically labeled compounds as defined herein, for example, compounds in which radioactive isotopes are present, such as ³ H and ¹⁴ C, or compounds in which non-radioactive isotopes are present, such as ² H and ¹³ C Such isotope-labeled compounds are useful in metabolic studies (using ¹⁴ C), reaction kinetics studies (using, for example, ² H or ³ H), detection or imaging techniques such as positron emission tomography (PET) or single photon emission computed tomography (SPECT), including drug or substrate tissue distribution assays, or in radiotherapy of patients. In particular, ^{the 18} F labeled compound may be particularly needed for PET or SPECT studies. Isotope-labeled compounds of formula (I) can generally be prepared by conventional techniques known to those skilled in the art, or by methods similar to those described in the accompanying examples, using suitable isotope-labeled reagents in place of the unlabeled reagents previously used.

In addition, substitution with heavier isotopes, particularly deuterium (i.e., ^2H or D), may provide certain therapeutic benefits due to greater resistance to metabolic inactivation, such as increased half-life in vivo, or decreased dosage requirements, or improvement in terms of the therapeutic index. It should be understood that deuterium is considered in this context as a substitute for the compound of formula (I). The concentration of such a heavier isotope, specifically deuterium, can be determined by means of an isotope enrichment factor. As used herein, the term "isotopic enrichment factor" means the relationship between the abundance of an isotope and the natural occurrence of that isotope. Where the substituent in a compound of the present invention is said deuterium, the compound has an isotopic enrichment factor for each designated deuterium atom of at least 3500 (52.5% deuterium input for each designated deuterium atom), at least 4000 ( deuterium injection 60%), at least 4500 (deuterium injection 67.5%), at least 5000 (deuterium injection 75%), at least 5500 (deuterium injection 82.5%), at least 6000 (deuterium injection 90%), at least 6333.3 (95% deuterium injection), at least 6466.7 (97% deuterium injection), at least 6600 (99% deuterium injection) or at least 6633.3 (deuterium injection 99.5%).

Pharmaceutically acceptable solvates in accordance with the present invention include those where the crystallization solvent may be replaced by an isotope, for example, D₂O,d₆–acetone, d₆–DMSO.

The compounds according to the present invention, i.e. compounds of formula (I) which contain groups capable of acting as hydrogen bond donors and/or acceptors may be capable of forming cocrystals with suitable cocrystal forming agents. Such cocrystals can be prepared from compounds of formula (I) by known cocrystal formation procedures. Such procedures include grinding, heating, co-sublimation, co-melting or contacting in solution the compounds of formula (I) with an agent to form co-crystals under crystallization conditions and isolating the co-crystals thus formed. Suitable means for forming cocrystals include those described in WO 2004/078163. Therefore, the present invention further provides cocrystals containing a compound of formula (I).

As used herein, the term "pharmaceutically acceptable carrier" includes any and all solvents, dispersion media, coatings, surfactants, antioxidants, preservatives (e.g. antibacterial agents, antifungal agents), isotonicity agents, absorption retardants, salts, preservatives, drug stabilizers, binders, excipients, disintegrants, lubricants, sweeteners, flavoring agents, coloring agents, etc. and combinations thereof, which will be known to those skilled in the art (see, for example, Remington's Pharmaceutical Sciences, 18^th Ed. Mack Printing Company, 1990, pp. 1289–1329). Unless any conventional carrier is incompatible with the active ingredient, it is intended for use in therapeutic or pharmaceutical compositions.

The term "therapeutically effective amount" of a compound of the present invention refers to an amount of a compound of the present invention that will produce a biological or medical response in a subject, such as reducing or inhibiting the activity of an enzyme or protein, or reducing the severity of symptoms, alleviating a condition, slowing or inhibiting the progression diseases, or prevent diseases, etc. In one non-limiting embodiment, the term "therapeutically effective amount" refers to an amount of a compound of the present invention that, when administered to a subject, is effective to (1) at least partially alleviate, inhibit, prevent, and/or reduce the severity of a condition or disorder, or a disease (i) mediated by ALK-2, or (ii) associated with ALK-2 activity, or (iii) characterized by activity (normal or abnormal) of ALK-2; or (2) reducing or suppressing ALK-2 activity; or (3) reduction or suppression of ALK–2 expression. In another non-limiting embodiment, the term “therapeutically effective amount” refers to an amount of a compound of the present invention that, when administered to a cell or tissue or non-cellular biological material or environment, is effective to at least partially reduce or inhibit ALK-2 activity ; or at least partially reducing or suppressing ALK-2 expression.

As used herein, the term “subject” refers to a human being.

As used herein, the term “suppress,” “suppress,” or “suppress” refers to the reduction or attenuation of a given condition, symptom, or disorder, or disease, or a significant reduction in the original activity of a biological activity or process.

As used herein, the term “treat,” “treat,” or “cure” any disease or disorder refers in one embodiment to reducing the severity of the disease or disorder (i.e., slowing or stopping or reducing the progression of the disease or at least one of its clinical symptoms). In another embodiment, “treat,” “treat,” or “treat” refers to alleviating or reducing the severity of at least one physical parameter, including those that may not be apparent to the patient. In yet another embodiment, “treat,” “treat,” or “treat” refers to modulating a disease or disorder either physically, (eg, stabilization of an obvious symptom), or physiologically (for example, stabilization of a physical parameter), or with the help of both. In yet another embodiment, “treat,” “treat,” or “treat” refers to preventing or delaying the occurrence, or development, or progression of a disease or disorder.

As used herein, a subject "needs" treatment if such subject would benefit biologically, medically, or in terms of quality of life as a result of such treatment.

As used herein, the singular and similar terms used in the context of the present invention (particularly in the context of the claims) are to be construed to cover both the singular and plural forms unless otherwise specified herein or there is no obvious contradiction in context.

All methods described herein can be performed in any suitable order unless otherwise indicated herein or otherwise clearly contradicted by context. Use of any and all examples or exemplary language (e.g. "such as") as used herein is intended to better explain the present invention only and is not intended to limit the scope of the present invention otherwise claimed.

Any asymmetric atom (eg, carbon or the like) of the compound(s) of the present invention may be in a racemic or enantiomerically enriched form, for example, in the ( R )–, ( S )–, or ( R,S )– configuration. In certain embodiments, each asymmetric atom has an enantiomeric excess of at least 50%, an enantiomeric excess of at least 60%, an enantiomeric excess of at least 70%, an enantiomeric excess of at least 80%, an enantiomeric excess , an enantiomeric excess of at least 95%, or an enantiomeric excess of at least 99%, in the ( R )- or ( S )-configuration. Substituents on atoms with unsaturated double bonds may, if possible, be in the cis- ( Z )- or trans- ( E )-form.

Accordingly, the compound of the present invention as used herein may be in the form of one of possible isomers, rotamers, atropisomers, tautomers, or mixtures thereof, e.g., substantially pure geometric ( cis- or trans- ) isomers, diastereomers, optical isomers (antipodes) ), racemates or mixtures thereof.

Any resulting mixtures of isomers can be separated on the basis of physicochemical differences between the constituents into pure or substantially pure geometric or optical isomers, diastereomers, racemates, for example, by chromatography and/or fractional crystallization.

Any resulting racemates of final products or intermediates can be separated into optical antipodes by known methods, e.g. by separating their diastereomeric salts obtained with an optically active acid or base and isolating the optically active acid or base compound. In particular, the basic fragment can thus be used to separate the compounds of the present invention into their optical antipodes, e.g. by fractional crystallization of a salt formed with an optically active acid, e.g. tartaric acid, dibenzoyltartaric acid, diacetyltartaric acid, di-O,O'–P-toluene tartaric acid, mandelic acid, malic acid or camphor-10-sulfonic acid. Racemic products can also be separated by chiral chromatography, e.g. high pressure liquid chromatography (HPLC) using a chiral adsorbent.

In addition, the compounds of the present invention, including their salts, can also be obtained in the form of their hydrates or include other solvents used for their crystallization. The compounds of the present invention may, by nature or by design, form solvates with pharmaceutically acceptable solvents (including water); therefore, the present invention is intended to cover both solvated and non-solvated forms. The term "solvate" refers to a molecular complex of a compound of the present invention (including its pharmaceutically acceptable salts) with one or more solvent molecules. Such solvent molecules are those commonly used in the pharmaceutical field which are known to be non-toxic to the recipient, such as water, ethanol and the like. The term "hydrate" refers to a complex in which the solvent molecule is water.

The compounds of the present invention, including their salts, hydrates and solvates, may, by nature or by design, form polymorphs.

In general, compounds of formula (I) can be prepared according to the schemes presented below.

Scheme 1

Step 1. A compound of formula (III), wherein R ₂ , R ₃ , R ₄ and L are defined herein with respect to a compound of formula (I), can be prepared by coupling a compound of formula (IV), wherein moiety B( OR) ₂ forms a boronic acid derivative (for example, where R is hydrogen or pinacol), with a compound of formula (V), wherein R ₂ , R ₃ , R ₄ and L are defined herein in relation to a compound of formula (I), in the presence of a suitable solvent, such as, for example, dioxane or 2-methyl-2-butanol, a base, such as potassium carbonate, and a suitable catalyst, preferably a palladium-based catalyst, such as, for example, [1,1'-bis( diphenylphosphino)ferrocene]dichloropalladium(II). The compounds of formulas IV, V, III and II can be used in free form or in the form of their salts.

Step 2: A compound of formula (II), wherein R ₂ , R ₃ , R ₄ and L are defined herein with respect to a compound of formula (I), can be prepared by treating a compound of formula (III), wherein R ₂ , R ₃ , R ₄ and L are defined herein with respect to a compound of formula (I), using a suitable base, such as, for example, lithium hydroxide or sodium hydroxide, in the presence of a suitable solvent, such as, for example, tetrahydrofuran or an alkanol such as methanol. Addition of an acid such as HCl results in the formation of the corresponding salt, for example the hydrochloride salt, of the compound of formula II.

Step 3: A compound of formula (I) wherein R ₁ , R ₂ , R ₃ , R ₄ and L are defined herein can be prepared by coupling a compound of formula (II) wherein R ₂ , R ₃ , R ₄ and L are defined herein in relation to a compound of formula (I), with a suitable amine having the formula R ₁ –NH ₂ , wherein R ₁ is defined herein in relation to a compound of formula (I), such as, for example, trans - 4-aminocyclohexanol, a suitable amide coupling reagent such as O-(7-azabenzotriazol-1-yl)-N,N,N´,N´-tetramethyluronium hexafluorophosphate, and a base such as N-methylmorpholine or triethylamine, in the presence of a suitable solvent such as, for example, N,N-dimethylformamide or acetonitrile.

The compounds of formulas (IV) and (V) can be prepared as described in the examples provided further below.

Scheme 2

Step 1. A compound of formula (VII), wherein R ₁ is defined herein in relation to a compound of formula (I), can be prepared by treating a compound of formula (VIII) using a suitable amine of formula R ₁ –NH ₂ such as, for example , trans -4-aminocyclohexanol, a suitable amide coupling reagent such as O-(7-azabenzotriazol-1-yl)-N,N,N´,N´-tetramethyluronium hexafluorophosphate, and a base such as , for example N-methylmorpholine, in the presence of a suitable solvent, such as, for example, N,N-dimethylformamide.

Step 2. A compound of formula (VI), wherein R ₁ is defined herein with respect to a compound of formula (I), and the B(OR) ₂ moiety forms a boronic acid derivative (eg, where R is hydrogen or pinacol), may be prepared by reacting a compound of formula (VII), wherein R ₁ is defined herein with respect to a compound of formula (I), with a boron compound, such as, for example, bis(pinacolato)diboron, in the presence of a suitable solvent, such as, for example, dioxane, a salt such as potassium acetate, and a suitable catalyst, preferably a palladium-based catalyst such as, for example, [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(II).

Step 3. A compound of formula (I), wherein R ₁ , R ₂ , R ₃ , R ₄ and L are defined herein, can be prepared by coupling a compound of formula (VI), wherein R ₁ is defined herein with respect to to a compound of formula (I), and the B(OR) ₂ moiety forms a boronic acid derivative (eg, where R is hydrogen or pinacol), with a compound of formula (V), wherein R ₂ , R ₃ , R ₄ and L are defined in herein with respect to a compound of formula (I), in the presence of a suitable solvent, such as, for example, dioxane, a base, such as sodium hydroxide, and a suitable catalyst, preferably a palladium-based catalyst, such as, for example, [1,1' –bis(diphenylphosphino)ferrocene]dichloropalladium(II).

Scheme 3

Step 1. A compound of formula (VIII), wherein R ₂ , R ₃ , L, R ₄ are defined herein with respect to a compound of formula (I), and the B(OR) ₂ moiety forms a boronic acid derivative (for example, where R is is hydrogen or pinacol) can be prepared by coupling a compound of formula (V), wherein R ₂ , R ₃ , L, R ₄ are defined herein with respect to a compound of formula (I), with a boron compound, such as for example, bis(pinacolato)diboron, in the presence of a suitable solvent, such as, for example, dioxane, a salt, such as potassium acetate, and a suitable catalyst, preferably a palladium-based catalyst, such as, for example, [1,1'-bis( diphenylphosphino)ferrocene]dichloropalladium(II), or alternatively in a suitable solvent such as tetrahydrofuran and a strong base such as n-butyllithium or isopropylmagnesium chloride, or a mixture of the two with a boron compound such as trimethyl boronate.

Step 2. A compound of formula (I), wherein R ₁ , R ₂ , R ₃ , R ₄ and L are defined herein, can be prepared by coupling a compound of formula (VIII), where R ₂ , R ₃ , L, R ₄ is defined herein with respect to a compound of formula (I), and the B(OR) ₂ moiety forms a boronic acid derivative (eg, where R is hydrogen or pinacol), with a compound of formula (VII), where R ₁ is defined in herein with respect to a compound of formula (I), in the presence of a suitable solvent, such as, for example, dioxane, a base, such as potassium carbonate, and a suitable catalyst, preferably a palladium-based catalyst, such as, for example, [1,1' –bis(diphenylphosphino)ferrocene]dichloropalladium(II).

Scheme 4

Step 1. A compound of formula (IX), wherein R ₂ and R ₃ are defined herein with respect to a compound of formula (I), can be prepared by treating a compound of formula (X), wherein R ₂ and R ₃ are defined herein with respect to in relation to the compound of formula (I), using N-methyliminodiacetic acid in the presence of a suitable solvent, such as, for example, DMF.

Step 2: A compound of formula (VIII-1), wherein R ₁ , R ₂ , R ₃ , R ₄ and L are defined herein, can be prepared by treating a compound of formula (IX), wherein R ₂ and R ₃ are defined herein document in relation to a compound of formula (I), using a suitable amine with a suitable reducing agent, such as, for example, sodium triacetoxyborohydride, and an acid, such as acetic acid, in the presence of a suitable solvent, such as, for example, THF.

Step 3. A compound of formula (I), wherein R ₁ , R ₂ , R ₃ , R ₄ and L are defined herein, can be prepared by coupling a compound of formula (VIII-1), where R ₂ , R ₃ , L, R ₄ are defined herein in relation to a compound of formula (I), with a compound of formula (VII), wherein R ₁ is defined herein in relation to a compound of formula (I), in the presence of a suitable solvent, such as, for example, dioxane, a base such as potassium phosphate, and a suitable catalyst, preferably a palladium-based catalyst such as (2-dicyclohexylphosphino-2′,4′,6′-triisopropyl-1,1′-biphenyl) chloride[2 –(2-aminoethyl)phenyl)]palladium(II).

In a further aspect, the present invention relates to a process for preparing a compound of formula (I) in free form or in pharmaceutically acceptable salt form, comprising the steps of:

a) reacting a compound of formula (II) as defined herein with a compound of formula R ₁ –NH ₂ as defined herein to produce a compound of formula (I);

b) isolating the compound of formula (I) thus obtained in free form or in the form of a pharmaceutically acceptable salt.

In a further aspect, the present invention relates to a process for preparing a compound of formula (I) in free form or in pharmaceutically acceptable salt form, comprising the steps of:

a) reacting a compound of formula (VI) as defined herein with a compound of formula (V) as defined herein to produce a compound of formula (I);

b) isolating the compound of formula (I) thus obtained in free form or in the form of a pharmaceutically acceptable salt.

In a further aspect, the present invention relates to a process for preparing a compound of formula (I) in free form or in pharmaceutically acceptable salt form, comprising the steps of:

a) reacting a compound of formula (VIII) or a compound of formula (VIII-1) as defined herein with a compound of formula (VII) as defined herein to produce a compound of formula (I);

b) isolating the compound of formula (I) thus obtained in free form or in the form of a pharmaceutically acceptable salt.

The present invention further includes any embodiment of the methods of the present invention in which the intermediate product obtained from any step thereof is used as a starting material and the remaining steps are carried out, or in which the starting materials are produced in situ under the reaction conditions, or in which the components of the reaction mixture used in the form of their salts or optically pure material.

The compounds of the present invention and intermediates can also be converted into each other in accordance with methods generally known to those skilled in the art.

In another aspect, the present invention provides a pharmaceutical composition comprising a compound of the present invention or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier. In a further embodiment, the composition contains at least two pharmaceutically acceptable carriers, such as those described herein. For purposes of the present invention, unless otherwise indicated, solvates and hydrates generally refer to compositions. Preferably, the pharmaceutically acceptable carriers are sterile. The pharmaceutical composition may be formulated for specific routes of administration, such as oral administration, parenteral administration, rectal administration, transdermal administration, etc. In addition, the pharmaceutical compositions of the present invention may be formulated in solid form (including, but not limited to, capsules, tablets, pills, granules, powders, or suppositories) or liquid form (including, but not limited to, solutions, suspensions, or emulsions). Pharmaceutical compositions may undergo standard pharmaceutical operations such as sterilization, and/or may contain standard inert diluents, lubricants or buffers, as well as adjuvants such as preservatives, stabilizers, wetting agents, emulsifiers and buffers, etc.

Typically, pharmaceutical compositions are tablets or gelatin capsules containing the active ingredient together with one or more of:

a) thinners, for example, lactose, dextrose, sucrose, mannitol, sorbitol, cellulose and/or glycine;

b) lubricants, e.g. silicon dioxide, talc, stearic acid, its magnesium or calcium salt and/or polyethylene glycol; also for tablets

c) binding agents, e.g. magnesium aluminosilicate, starch paste, gelatin, tragacanth, methylcellulose, sodium carboxymethylcellulose and/or polyvinylpyrrolidone; if necessary

d) raising agents, e.g. types of starch, agar, alginic acid or its sodium salt or effervescent mixtures; And

e) absorbents, dyes, flavors and sweeteners.

Tablets can be either film-coated or enteric-coated according to methods known in the art.

Suitable compositions for oral administration include an effective amount of a compound of the present invention in the form of tablets, lozenges, aqueous or oily suspensions, dispersible powders or granules, emulsions, hard or soft capsules, or syrups or tinctures. Compositions intended for oral administration are prepared in accordance with any method known in the art for the manufacture of pharmaceutical compositions, and such compositions may contain one or more agents selected from the group consisting of sweeteners, flavoring agents, coloring agents and preservatives. means, with the aim of obtaining drugs that are pharmaceutically acceptable and have an attractive taste. Tablets may contain the active ingredient in admixture with non-toxic pharmaceutically acceptable excipients that are suitable for the manufacture of tablets. Such excipients are, for example, inert diluents such as calcium carbonate, sodium carbonate, lactose, calcium phosphate or sodium phosphate; granulating and disintegrating agents such as corn starch or alginic acid; binders such as starch, gelatin or gum arabic; and lubricants such as magnesium stearate, stearic acid or talc. The tablets are uncoated or coated by known techniques to delay disintegration and absorption in the gastrointestinal tract and thereby provide sustained action over a longer period. For example, a delayed release material such as glyceryl monostearate or glyceryl distearate may be used. Formulations for oral administration may be presented as hard gelatin capsules, where the active ingredient is mixed with an inert solid diluent, for example, calcium carbonate, calcium phosphate or kaolin, or as soft gelatin capsules, where the active ingredient is mixed with water or an oily vehicle, for example , peanut oil, liquid paraffin or olive oil.

The compounds of formula (I), in free form or in pharmaceutically acceptable salt form, exhibit valuable pharmacological properties, for example ALK-2-modulating properties, for example shown by the in vitro and in vivo assays presented in the following sections, and are therefore intended for therapy or for use as research chemicals, for example as pharmacologically active compounds.

The compounds of the present invention may be useful in the treatment of an indication selected from heterotopic ossification or fibrodysplasia ossificans progressiva.

Without being limited by theory, it is believed that the compounds of the present invention, which are selective inhibitors of ALK-2, reduce/suppress BMP signaling and the repair of abnormal tissue associated with it.

Thus, as a further embodiment, the present invention provides for the use of a compound of formula (I) or its subformulas (Ia), (II), (IIa), (IIb) in free form or in pharmaceutically acceptable salt form in therapy. In a further embodiment, the therapy is selected from a disease that can be treated by inhibiting the ALK-2 receptor. In another embodiment, the disease is selected from heterotopic ossification or fibrodysplasia ossificans progressiva.

Thus, as a further embodiment, the present invention provides a compound of formula (I) or subformulas (Ia), (II), (IIa), (IIb) in free form or in pharmaceutically acceptable salt form for use in therapy. In a further embodiment, the therapy is selected from a disease that can be treated by inhibiting the ALK-2 receptor. In another embodiment, the disease is selected from heterotopic ossification or fibrodysplasia ossificans progressiva.

In another embodiment, the present invention provides a method of treating a disease treated by inhibiting the ALK-2 receptor, comprising administering a therapeutically acceptable amount of a compound of formula (I) or subformulas (Ia), (II), (IIa), (IIb) thereof. in free form or in the form of a pharmaceutically acceptable salt. In a further embodiment, the disease is selected from heterotopic ossification or fibrodysplasia ossificans progressiva.

Thus, as a further embodiment, the present invention provides for the use of a compound of formula (I) or its subformulas (Ia), (II), (IIa), (IIb) in free form or in pharmaceutically acceptable salt form for the manufacture of a medicinal product. In a further embodiment, the medicinal product is for the treatment of a disease that can be treated by inhibiting the ALK-2 receptor. In another embodiment, the disease is selected from heterotopic ossification or fibrodysplasia ossificans progressiva.

In one embodiment of the present invention, 2-amino-N-(4-hydroxybicyclo[2.2.2]octan-1-yl)-5-(4-(3-(tetrahydro-2H-pyran-4-yl)-3 -azabicyclo[3.1.0]hexan-1-yl)phenyl)nicotinamide in free form or a pharmaceutically acceptable salt thereof, in particular in one of the modifications specified herein, for use in the treatment of heterotopic ossification or fibrodysplasia ossificans progressiva.

In one embodiment, 2-amino-N-(4-hydroxybicyclo[2.2.2]octan-1-yl)-5-(4-((1R,5S)-3-(tetrahydro-2H-pyran-4- yl)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)nicotinamide in free form or a pharmaceutically acceptable salt thereof, in particular in one of the modifications specified herein, for use in the treatment of heterotopic ossification or progressive fibrodysplasia ossificans.

In one embodiment, 2-amino-N-(4-hydroxybicyclo[2.2.2]octan-1-yl)-5-(4-((1S,5R)-3-(tetrahydro-2H-pyran-4- yl)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)nicotinamide in free form or a pharmaceutically acceptable salt thereof, in particular in one of the modifications specified herein, for use in the treatment of heterotopic ossification or progressive fibrodysplasia ossificans

The following examples are intended to illustrate the present invention and should not be construed as limiting it. Temperature values are given in degrees Celsius. Unless otherwise specified, all evaporation operations are carried out under reduced pressure, typically from about 15 mmHg. up to 100 mmHg (=20–133 mbar). The structures of the final products, intermediates and starting materials were confirmed by standard analytical methods, e.g. microanalysis and spectroscopic characteristics, eg MS, IR, NMR. The abbreviations used are standard prior art abbreviations.

All starting materials, building blocks, reagents, acids, bases, dehydrogenating agents, solvents and catalysts used for the synthesis of the compounds of the present invention are either commercially available or can be prepared by organic synthesis methods known to one skilled in the art, or through methods described or similar, respectively, to the methods presented in the Examples section. Also, the compounds of the present invention can be prepared by organic synthesis methods known to one skilled in the art, as shown in the following examples.

Examples

Abbreviations

δ chemical shift AcOH acetic acid water or aq. water APCI–MS chemical ionization with mass spectrometry at atmospheric pressure BH ₃ DMS borane–dimethyl sulfide complex Saline solution saturated sodium chloride solution n–BuLi n-butyllithium DCM dichloromethane DMAP 4-(dimethylamino)pyridine DMF N,N–dimethylformamide DIPEA diisopropylethylamine DMSO dimethyl sulfoxide DSC differential scanning calorimetry DVS dynamic vapor sorption eq. equivalent(s) Et ethyl Et _3N triethylamine EtOAc ethyl acetate EtOH ethanol ESI–MS electrospray ionization with mass spectrometry FIA–MS flow injection analysis with mass spectrometry FT–IR Fourier transform infrared spectroscopy h hour HATU O–(7–azabenzotriazol–1–yl)–N,N,N′,N′–tetramethyluronium hexafluorophosphate HPLC high performance liquid chromatography HRMS high resolution mass spectrometry IPA isopropanol IPAc isopropyl acetate i –PrOH isopropanol IT internal temperature _{K2CO3 _} potassium carbonate _{K3PO4 _} potassium phosphate KOAc potassium acetate l liter LC–MS liquid chromatography with mass spectrometry _LiAlH4 lithium aluminum hydride LiOH lithium hydroxide M molar MCC microcrystalline cellulose mg milligram mm millimolar MeOH methanol min. minutes ml milliliter _MgSO4 magnesium sulfate MHz megahertz MTBE methyl tert-butyl ether n. normal _{Na2SO4 _} sodium sulfate _NaHCO3 bicarbonate of soda NaHMDS sodium bis(trimethylsilyl)amide NaOH sodium hydroxide _NH4Cl ammonium chloride _NH4OH ammonium hydroxide _NH4OAc ammonium acetate NMR nuclear magnetic resonance _PdCl2 (dppf) or Pd(dppf)Cl ₂ [1,1'–bis(diphenylphosphino)ferrocene]dichloropalladium(II) PdCl ₂ (PPh ₃ ) ₂ bis(triphenylphosphine)palladium(II) chloride prepHPLC preparative high performance liquid chromatography ppm parts per million RT or k.t. room temperature (23±3°C) sat. saturated SFC supercritical fluid chromatography SPE solid phase extraction TEA triethylamine T.F.A. trifluoroacetic acid THF tetrahydrofuran TGA thermogravimetric analysis t _R retention time UPLC–MS ultra-high performance liquid chromatography with mass spectrometry XPhos Pd (2-dicyclohexylphosphino-2′,4′,6′-triisopropyl-1,1′-biphenyl)[2-(2-aminoethyl)phenyl)]palladium(II) chloride XRPD powder x-ray diffraction

ANALYTICAL METHODS

1H-NMR

Measurements were performed on a Bruker Ultrashield ^TM 400 (400 MHz), Bruker Ultrashield ^TM 600 (600 MHz), 400 MHz DRX Bruker CryoProbe (400 MHz) or 500 MHz DRX Bruker CryoProbe (500 MHz) spectrometer, with or without trimethylsilane as an internal standard. him. Chemical shifts (δ-values) are given in ppm, counting from tetramethylsilane, the spectra in the splitting diagram are designated as singlet ( s ), doublet ( d ), triplet ( t ), quartet ( q ), multiplet, unseparated or more overlapping signals ( m ), broad signal ( bs ). Solvents are given in parentheses.

UPLC–MS

Column: Waters Acquity HSS T3, C18, 1.8 µm, 2.1 x 50 mm, oven at 60°C. Flow rate: 1.0 ml/min. Gradient: 5% to 98% B in 1.40 min, then 98% B in 0.40 min, 98% to 5% B in 0.10 min, 5% B in 0.10 min; A=water+0.05% formic acid+3.75 mM NH ₄ OAc, B=acetonitrile+0.04% formic acid. UV/VIS detection (DAD), ESI (+/−). Mass spectrometer range: 100–1200 Da.

In the case of examples 46, 56, 74, 81, 85 and 87:

column: Waters Acquity BEH, C18, 1.7 µm, 2.1x50 mm, oven at 50°C. Flow rate: 1.0 ml/min. Gradient: 2% to 98% B in 4.40 min, then 98% B in 0.75 min, 98% to 2% B in 0.04 min; A=water+0.1% formic acid, B=acetonitrile+0.1% formic acid. UV/VIS detection (DAD), ESI (+/−). Mass spectrometer range: 100–1200 Da.

UPLC–HRMS:

Waters Acquity SDS, C18, 1.7 µm, 2.1 x 50 mm, thermostat at 50°C. Gradient: 5% to 98% B in 7.50 min, then 98% B in 0.40 min, 98% to 5% B in 0.15 min; A=water+5 mM NH4OH, B=acetonitrile+5 mM NH4OH. UV/VIS detection (DAD), ESI (+/−). Mass spectrometer range: 100–1200 Da.

HPLC–MS

Column: Waters Symmetry C8, 3.5 µm, 2.1 x 50 mm, oven at 50°C. Flow rate: 1.0 ml/min.

Gradient: 10% to 95% B in 2 min, then 95% B in 1 min, 95% to 10% B in 0.5 min. 10% B for 0.50 min; A=water+0.1% TFA, B=acetonitrile+0.1% TFA. UV/VIS detection (DAD), APCI (+). Mass spectrometer range: 100–1200 Da.

Measuring the amount of rotation of the plane of polarization of light

The amount of rotation of the plane of polarization of light was measured using a Perkin Elmer series PE241 polarimeter No. 5325 operating at 589 nm using chloroform as a solvent.

CLEANING METHODS

prepHPLC

Gilson GX-281, pumps 331/332.

Column: Waters Sunfire C18, 30x100 mm, 5 µm. Flow rate: 30 ml/min.

Mobile phase: water (containing 0.1% TFA) and acetonitrile (method 1a).

Or:

Column: X-Bridge C18, 30x50 mm, 5 µm. Flow rate: 75 ml/min.

Mobile phase: water (containing 5 mM NH ₄ OH) and acetonitrile (method 1b).

Flash chromatography with normal phase

Teledyne ISCO CombiFlash

Column: Redisep Rf for flash chromatography with silica gel.

Mobile phase: cyclohexane/EtOAc (method 2a) or DCM/MeOH (method 2b).

Biotage Flash–Master II

Column: Pre-packed with silica gel 60 (40-63 µm), from Merck.

Mobile phase: DCM and MeOH (containing 7.3 mM NH ₄ OH) (method 2c).

Reverse phase flash chromatography

Teledyne ISCO CombiFlash

Column: Redisep Rf Gold C18 HPLC, 15.5g, 50g or 240g pre-packed columns, 20-40µm, 100A.

Mobile phase: water and acetonitrile (containing 7.3 mM NH ₄ OH) (method 3a) or water (containing 0.1% TFA) and acetonitrile (method 3b).

Example 1 : 2-amino-N-((1r,4R)-4-hydroxycyclohexyl)-5-(4-((1R,5S)-3-(1-(methylsulfonyl)piperidin-4-yl)-3- azabicyclo[3.1.0]hexane–1–yl)phenyl)nicotinamide

Into a solution of TFA salt and 5-(4-((1R,5S)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)-2-amino-N-((1r,4R)-4-hydroxycyclohexyl )nicotinamide (intermediate 1a, 80 mg, 0.158 mmol) in DCM (4 ml) was added 1-(methylsulfonyl)piperidin-4-one (30.8 mg, 0.174 mmol) and AcOH (0.018 ml, 0.316 mmol) under atmosphere nitrogen at room temperature The reaction mixture was stirred at 45°C for 30 minutes. Sodium triacetoxyborohydride (84 mg, 0.395 mmol) was added at room temperature. and the reaction mixture was stirred at 45°C for 1 hour. The reaction mixture was then diluted with sat. aq. NaHCO ₃ solution and mixed with EtOAc. After phase separation aq. the layer was again extracted with EtOAc. The combined organic layers were washed with brine, dried over MgSO ₄ , filtered and concentrated under reduced pressure. The crude product was purified by normal phase chromatography (method 2b) to obtain the title compound as a colorless solid. ^1H NMR (400 MHz, DMSO–d6) δ 8.39 (d, 1H), 8.31 (d, 1H), 8.11 (d, 1H), 7.59 (d, 2H), 7. 24 (d, 2H), 7.12 (s, 2H), 4.58 (d, 1H), 3.80–3.63 (m, 1H), 3.45–3.36 (m, 4H) , 3.15 (dd, 1H), 2.89–2.83 (m, 5H), 2.65 (dd, 1H), 2.35–2.29 (m, 1H), 1.98–1 .81 (m, 7H), 1.57–1.18 (m, 8H), 0.83–0.70 (m, 1H). (UPLC–MS) t _R 0.49 min; ESI–MS 554 [M+H] ⁺ .

Intermediate 1a : 5-(4-((1R,5S)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)-2-amino-N-((1r,4R)-4-hydroxycyclohexyl) nicotinamide

Into a solution of (1R,5S) -tert -butyl-1-(4-(6-amino-5-(((1r,4R)-4-hydroxycyclohexyl)carbamoyl)pyridin-3-yl)phenyl)-3-azabicyclo [3.1.0]hexane-3-carboxylate (intermediate 1b, 940 mg, 1.55 mmol) in dioxane (10 ml) was added 4 M HCl in dioxane (1.55 ml, 6.20 mmol) at rt. T. The reaction mixture was stirred at 65°C for 5 hours and then concentrated under reduced pressure to obtain the title compound as the hydrochloride salt. The optional title compound was further purified by prepHPLC (Method 1a) to obtain the TFA salt after evaporation of the solvents. (UPLC–MS) t _R 0.45 min; ESI–MS 393 [M+H] ⁺ .

Intermediate 1b : (1R,5S) -tert -butyl-1-(4-(6-amino-5-(((1r,4R)-4-hydroxycyclohexyl)carbamoyl)pyridin-3-yl)phenyl)-3 –azabicyclo[3.1.0]hexane-3-carboxylate

Into a solution of 2-amino-5-(4-((1R,5S)-3-( tert -butoxycarbonyl)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)nicotinic acid (intermediate 1c, 630 mg, 1.59 mmol) in DMF (10 ml) was added trans -4-aminocyclohexanol (290 mg, 1.91 mmol), HATU (909 mg, 2.39 mmol) and N-methylmorpholine (0.53 ml, 4 .78 mmol) at room temperature The reaction mixture was stirred for 2 hours, diluted with sat. aq. NaHCO ₃ solution and extracted three times with EtOAc. The combined organic layers were washed with brine, dried over MgSO ₄ , filtered and concentrated under reduced pressure. The crude product was purified by prepHPLC (Method 1a) to obtain the title compound as a colorless solid. (UPLC–MS) t _R 1.00 min; ESI–MS 493 [M+H] ⁺ .

Intermediate 1c : 2-amino-5-(4-((1R,5S)-3-( tert -butoxycarbonyl)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)nicotinic acid

Into a solution of (1R,5S) -tert -butyl-1-(4-(6-amino-5-(methoxycarbonyl)pyridin-3-yl)phenyl)-3-azabicyclo[3.1.0]hexane-3-carboxylate ( intermediate 1d, 1.00 g, 2.44 mmol) in THF (15 ml) was added 2 M aq. LiOH solution (3.66 ml, 7.33 mmol) at RT. The reaction mixture was stirred at 65°C for 2 hours and then concentrated under reduced pressure. The crude product was purified by prepHPLC (Method 1a) to give the title compound. (UPLC–MS) t _R 0.92 min; ESI–MS 396 [M+H] ⁺ .

Intermediate 1d : (1R,5S) -tert -butyl-1-(4-(6-amino-5-(methoxycarbonyl)pyridin-3-yl)phenyl)-3-azabicyclo[3.1.0]hexane-3- carboxylate

Racemic tert -butyl-1-(4-(6-amino-5-(methoxycarbonyl)pyridin-3-yl)phenyl)-3-azabicyclo[3.1.0]hexane-3-carboxylate (intermediate 1e, 13) was separated 5 g, 33.0 mmol) using chiral preparative SFC (Waters SFC 200, Waters SFC 200, CHIRALPAK AD–H 5 µm, 30 * 250 mm, mobile phase CO ₂ /i–PrOH, 70:30, flow rate 120 g/min; UV detection at 278 nm). The title compound was obtained as an off-white solid after concentration under reduced pressure. Chiral analytical SFC (CHIRALPAK AD–3 3 µm, 2 * 100 mm, mobile phase CO ₂ /MeOH+0.1% DEA, 60:40, flow rate 1 ml/min; UV detection at 274 nm): t _R 2.30 min, ee=98.6%, [α] _D ²⁰ = +90°.

Intermediate 1e : tert -butyl-1-(4-(6-amino-5-(methoxycarbonyl)pyridin-3-yl)phenyl)-3-azabicyclo[3.1.0]hexane-3-carboxylate

Into a solution of tert -butyl-1-(4-bromophenyl)-3-azabicyclo[3.1.0]hexane-3-carboxylate (prepared as described in WO 2007/022935) (3.34 g, 9.38 mmol) in dioxane (75 ml) methyl 2-amino-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)nicotinate (prepared as described in WO 2012/087938) (3, 91 g, 14.1 mmol). 2 M aq was added. a solution of K ₂ CO ₃ (9.38 ml, 18.8 mmol) and the adduct of Pd(dppf)Cl ₂ and DCM (0.766 g, 0.938 mmol) under nitrogen and the mixture was heated to 80°C under nitrogen for 18 h The reaction mixture was diluted with 150 ml of water and extracted three times with EtOAc. The combined organic layers were washed with brine, dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure. The crude residue was purified by normal phase chromatography (method 2b) to give the title compound as an off-white solid. (UPLC–MS) t _R 1.29 min; ESI–MS 410 [M+H] ⁺ .

Example 2 : 2-amino-N-((1r,4R)-4-hydroxycyclohexyl)-5-(4-((1R,5S)-3-isopropyl-3-azabicyclo[3.1.0]hexan-1-yl )phenyl)nicotinamide

Into a solution of TFA salt and 2-amino-5-(4-((1R,5S)-3-isopropyl-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)nicotinic acid (intermediate 2a, 117 mg , 0.259 mmol) in DMF (3 ml), trans -4-aminocyclohexanol hydrochloride (39.3 mg, 0.259 mmol), HATU (148 mg, 0.389 mmol) and N-methylmorpholine (0.085 ml, 0.778 mmol) were added at room temperature . The reaction mixture was stirred at room temperature. for 2 hours and then diluted with sat. aq. NaHCO ₃ solution and mixed with EtOAc. After phase separation aq. the layer was again extracted with EtOAc. The combined organic layers were washed with brine, dried over MgSO ₄ , filtered and concentrated under reduced pressure. The crude product was purified by prepHPLC (Method 1a) to obtain the title compound as a yellow solid. 1H NMR (400 MHz, DMSO–d6) δ 8.38 (d,1H), 8.31 (d, 1H), 8.11 (d, 1H), 7.58 (d, 2H), 7.23 (d, 2H), 7.12 (s, 2H), 4.58 (d, 1H), 3.82–3.66 (m, 1H), 3.48–3.34 (m, 3H), 3.07 (d, 1H), 2.64–2.55 (m, 2H), 1.96–1.77 (m, 5H), 1.50–1.19 (m, 5H), 1, 04 (dd, 6H), 0.75 (dd, 1H). (UPLC–MS) t _R 0.50 min; ESI–MS 435 [M+H] ⁺ .

Intermediate 2a : TFA salt of 2-amino-5-(4-((1R,5S)-3-isopropyl-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)nicotinic acid

Into a solution of methyl 2-amino-5-(4-((1R,5S)-3-isopropyl-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)nicotinate (intermediate 2b, 660 mg, 1 .22 mmol) in THF (10 ml) was added 2 M aq. LiOH solution (1.22 ml, 2.44 mmol) at RT. The reaction mixture was stirred at 65°C for 2 hours and then concentrated under reduced pressure. The crude product was purified by prepHPLC (Method 1a) to obtain the title compound as the TFA salt. (UPLC–MS) t _R 0.37 min; ESI–MS 338 [M+H] ⁺ .

Intermediate 2b : Methyl-2-amino-5-(4-((1R,5S)-3-isopropyl-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)nicotinate

Into a solution of methyl 2-amino-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)nicotinate (prepared as described in WO 2012/087938) (462 mg, 1.663 mmol ) in acetonitrile (7 ml) was added (1R,5S)-1-(4-bromophenyl)-3-isopropyl-3-azabicyclo[3.1.0]hexane (intermediate 2c, 466 mg, 1.66 mmol), 2 M aq. K ₂ CO ₃ (1.66 ml, 3.33 mmol) and PdCl ₂ adduct (dppf) and DCM (67.9 mg, 0.083 mmol) at RT. The reaction mixture was capped and irradiated in a microwave reactor at 120°C for 10 min, then cooled, filtered through celite, diluted with sat. aq. NaHCO ₃ solution and mixed with EtOAc. After phase separation aq. the layer was again extracted with EtOAc. The combined organic layers were washed with brine, dried over MgSO ₄ , filtered and concentrated under reduced pressure. The crude product was used without further purification. (UPLC–MS) t _R 0.62 min; ESI–MS 352 [M+H] ⁺ .

Intermediate 2c : (1R,5S)–1–(4-bromophenyl)–3-isopropyl-3-azabicyclo[3.1.0]hexane

Into a solution of (1R,5S)-1-(4-bromophenyl)-3-azabicyclo[3.1.0]hexane (prepared as described in WO 2007/022935) (525 mg, 2.21 mmol) in acetonitrile (10 ml) 2-iodopropane (0.420 ml, 2.65 mmol) and K ₂ CO ₃ (609 mg, 4.41 mmol) were added at room temperature. in a nitrogen atmosphere. The reaction mixture was stirred at 65°C for 3 hours, then diluted with EtOAc and mixed with sat. aq. NaHCO ₃ solution. After phase separation aq. the phase was extracted with EtOAc. The combined organic layers were washed with brine, dried over MgSO ₄ , filtered and concentrated under reduced pressure to give the title compound as an off-white solid. (UPLC–MS) t _R 0.72 min; ESI–MS 280/282 [M+H] ⁺ .

Example 3 : 5-(4-((R)-1-(2-oxa-6-azaspiro[3.3]heptan-6-yl)ethyl)phenyl)-2-amino-N-((1r,4R)- 4-hydroxycyclohexyl)nicotinamide

To a solution of (6-amino-5-(((1r,4r)-4-hydroxycyclohexyl)carbamoyl)pyridin-3-yl)boronic acid (intermediate 3a, 300 mg, 0.645 mmol) in dioxane (5 ml) was added ( R)-6-(1-(4-bromophenyl)ethyl)-2-oxa-6-azaspiro[3.3]heptane (intermediate 3b, 420 mg, 0.774 mmol), PdCl ₂ (dppf) (23.6 mg, 0.032 mmol) and 2 n. aq. NaOH (0.645 ml, 1.29 mmol) under nitrogen at room temperature. The reaction mixture was stirred at 80°C for 2 hours, then diluted with EtOAc and water. After two extraction steps with EtOAc, the organic layers were washed with sat. aq. NaHCO ₃ solution and saline solution, respectively, dried over MgSO ₄ , filtered and concentrated under reduced pressure. The crude product was dissolved in MeOH and passed through a silica gel thiol cartridge (500 mg load; preloaded with MeOH) to remove palladium. The filtrate was concentrated under reduced pressure. The residue was purified by normal phase chromatography (method 2b) to give the title compound as a brownish solid. ^1H NMR (400 MHz, DMSO–d6) δ 8.37 (d, 1H), 8.27 (d, 1H), 8.09–8.14 (m, 1H), 7.57 (bs, 2H ), 7.33 (bs, 2H), 7.12 (bs, 2H), 4.54–4.58 (m, 1H), 4.51–4.66 (m, 3H), 3.55– 3.81 (m, 1H), 3.35–3.54 (m, 2H), 3.20–3.24 (m, 1H), 3.20–3.27 (m, 2H), 3, 15 (bs, 1H), 1.78–1.91 (m, 4H), 1.14–1.41 (m, 5H), 1.11 (bs, 3H). (UPLC–MS) t _R 0.45 min; ESI–MS 437 [M+H] ⁺ .

Intermediate 3a : (6-amino-5-(((1r,4r)-4-hydroxycyclohexyl)carbamoyl)pyridin-3-yl)boronic acid.

To a solution of 2-amino-5-bromo-N-((1r,4r)-4-hydroxycyclohexyl)nicotinamide (intermediate 3c, 1.00 g, 2.67 mmol) in dioxane (25 ml) was added bis(pinacolato) diboron (0.815 g, 3.21 mmol), PdCl ₂ (dppf) (0.098 g, 0.134 mmol) and KOAc (0.525 g, 5.35 mmol) under nitrogen at RT. The reaction mixture was stirred at 80°C for 18 hours, cooled, filtered through a pad of celite and concentrated under reduced pressure to give the title compound (the pinacolate ester was hydrolyzed under the reaction conditions) as a crude brownish solid, which was used without further purification . (UPLC–MS) t _R 0.32 min; ESI–MS 280 [M+H] ⁺ .

Intermediate 3b : (R)-6-(1-(4-bromophenyl)ethyl)-2-oxa-6-azaspiro[3.3]heptane.

To a solution of (R)-1-(4-bromophenyl)ethanamine (0.216 ml, 1.499 mmol) in DMF (7 ml) was added DIPEA (0.524 ml, 3.00 mmol) and 3,3-bis(bromomethyl)oxetane (439 mg, 1.80 mmol) in a nitrogen atmosphere at room temperature. The reaction mixture was stirred at 100°C for 5 hours, cooled and diluted with water and EtOAc. After phase separation, the aqueous layer was extracted twice with EtOAc and the combined organic layers were washed with brine, dried over MgSO ₄ , filtered and concentrated under reduced pressure to give the title compound as a yellow oil, which was used without further purification. (UPLC–MS) t _R 0.52 min; ESI–MS 282/284 [M+H] ⁺ .

Intermediate 3c : 2-amino-5-bromo-N-((1r,4r)-4-hydroxycyclohexyl)nicotinamide.

To a solution of 2-amino-5-bromonicotinic acid (2.00 g, 9.22 mmol) in DMF (30 ml) was added trans -4-aminocyclohexanol (1.68 g, 11.1 mmol), HATU (7.01 g, 18.4 mmol) and N-methylmorpholine (4.05 ml, 36.9 mmol) at room temperature. After stirring for 2 hours, the reaction mixture was diluted with EtOAc and sat. aq. NaHCO ₃ solution. The aqueous phase was extracted twice with EtOAc. The combined organic layers were dried over MgSO ₄ , filtered and concentrated under reduced pressure. The residue was purified by normal phase chromatography (method 2a) to give the title compound as a yellow solid. (UPLC–MS) t _R 0.62 min; ESI–MS 314/316 [M+H] ⁺ .

Example 4 : 5-(4-((S)-1-(2-oxa-6-azaspiro[3.3]heptan-6-yl)ethyl)phenyl)-2-amino-N-((1r,4R)- 4-hydroxycyclohexyl)nicotinamide

The title compound was prepared in a manner similar to that for 5-(4-((R)-1-(2-oxa-6-azaspiro[3.3]heptan-6-yl)ethyl)phenyl)-2-amino-N-( (1r,4R)-4-hydroxycyclohexyl)nicotinamide (Example 3), except that (S)-6-(1-(4-bromophenyl)ethyl)-2-oxa-6-azaspiro[3.3]heptane was used (intermediate 4a) instead of (R)-6-(1-(4-bromophenyl)ethyl)-2-oxa-6-azaspiro[3.3]heptane (intermediate 3b). ^1H NMR (400 MHz, DMSO–d6) δ 8.37 (d, 1H), 8.27 (d, 1H), 8.09–8.14 (m, 1H), 7.57 (bs, 2H ), 7.33 (bs, 2H), 7.12 (bs, 2H), 4.54–4.58 (m, 1H), 4.51–4.66 (m, 3H), 3.55– 3.81 (m, 1H), 3.35–3.54 (m, 2H), 3.20–3.24 (m, 1H), 3.20–3.27 (m, 2H), 3, 15 (bs, 1H), 1.78–1.91 (m, 4H), 1.14–1.41 (m, 5H), 1.11 (bs, 3H). (UPLC–MS) t _R 0.45 min; ESI–MS 437 [M+H] ⁺ .

Intermediate 4a : (S)-6-(1-(4-bromophenyl)ethyl)-2-oxa-6-azaspiro[3.3]heptane

The title compound was prepared in a manner similar to (R)-6-(1-(4-bromophenyl)ethyl)-2-oxa-6-azaspiro[3.3]heptane (intermediate 3b), except that ( S)-1-(4-bromophenyl)ethanamine instead of (R)-1-(4-bromophenyl)ethanamine. (UPLC–MS) t _R 0.55 min; ESI–MS 282/284 [M+H] ⁺ .

Example 5 : 2-amino-N-((1r,4S)-4-hydroxycyclohexyl)-5-(4-((1S,5R)-3-(3-morpholinopropyl)-3-azabicyclo[3.1.0]hexane –1-yl)phenyl)nicotinamide

Into a solution of TFA salt and 5-(4-((1S,5R)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)-2-amino-N-((1r,4S)-4-hydroxycyclohexyl )nicotinamide (intermediate 5a, 70 mg, 0.138 mmol) in acetonitrile (3 ml) was added K ₂ CO ₃ (47.7 mg, 0.345 mmol) and 4-(3-bromopropyl)morpholine (47.9 mg, 0.166 mmol ) at k.t. The reaction mixture was stirred at 65°C for 2 hours. The reaction mixture was then diluted with sat. aq. NaHCO ₃ solution and mixed with EtOAc. After phase separation aq. the layer was again extracted with EtOAc. The combined organic layers were washed with brine, dried over MgSO ₄ , filtered and concentrated under reduced pressure. The crude product was purified by prepHPLC (Method 1a) to obtain the title compound as a yellow solid. ^1H NMR (400 MHz, DMSO–d6) δ 8.38 (d, 1H), 8.31 (d, 1H), 8.11 (d, 1H), 7.58 (d, 2H), 7. 22 (d, 2H), 7.12 (s, 2H), 4.58 (d, 1H), 3.82–3.67 (m, 1H), 3.61–3.50 (m, 4H) , 3.41 (s, 2H), 3.05 (d, 1H), 2.48–2.24 (m, 9H), 1.93–1.76 (m, 5H), 1.61 (m , 2H), 1.46–1.17 (m, 6H), 0.76 (dd, 1H). (UPLC–MS) t _R 0.40 min; ESI–MS 520 [M+H] ⁺ .

Intermediate 5a : 5–(4–((1S,5R)–3–azabicyclo[3.1.0]hexan–1–yl)phenyl)–2–amino–N–((1r,4R)–4-hydroxycyclohexyl) nicotinamide

In solution (1S,5R)– tert -butyl–1–(4–(6–amino–5–(((1r,4S)–4–hydroxycyclohexyl)carbamoyl)pyridin–3–yl)phenyl)–3–azabicyclo [3.1.0]hexane-3-carboxylate (intermediate 5b, 1.18 g, 1.95 mmol) in dioxane (10 ml) was added 4 M HCl in dioxane (1.95 ml, 7.78 mmol) at k.t. The reaction mixture was stirred at 65°C for 5 hours and then concentrated under reduced pressure to obtain the title compound as the hydrochloride salt. Optionally, the product was further purified by prepHPLC (Method 1a) to obtain the title compound as the TFA salt after evaporation of the solvents. (UPLC–MS) t _R 0.45 min; ESI–MS 393 [M+H] ⁺ .

Intermediate 5b : (1S,5R) -tert -butyl-1-(4-(6-amino-5-(((1r,4S)-4-hydroxycyclohexyl)carbamoyl)pyridin-3-yl)phenyl)-3 –azabicyclo[3.1.0]hexane-3-carboxylate

The title compound was prepared in a manner similar to (1R,5S) -tert -butyl-1-(4-(6-amino-5-(((1r,4R)-4-hydroxycyclohexyl)carbamoyl)pyridin-3-yl )phenyl)-3-azabicyclo[3.1.0]hexane-3-carboxylate (intermediate 1b), except that 2-amino-5-(4-((1S,5R)-3-( tert - butoxycarbonyl)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)nicotinic acid (intermediate 5c) instead of 2-amino-5-(4-((1R,5S)-3-( tert -butoxycarbonyl) –3-azabicyclo[3.1.0]hexan-1-yl)phenyl)nicotinic acid (intermediate 1c). (UPLC–MS) t _R 1.00 min; ESI–MS 493 [M+H] ⁺ .

Intermediate 5c : 2-amino-5-(4-((1S,5R)-3-( tert -butoxycarbonyl)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)nicotinic acid

The title compound was prepared in a manner similar to that for 2-amino-5-(4-((1R,5S)-3-( tert -butoxycarbonyl)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)nicotinic acid (intermediate 1c), except that (1S,5R) -tert -butyl-1-(4-(6-amino-5-(methoxycarbonyl)pyridin-3-yl)phenyl)-3-azabicyclo was used [3.1.0]hexane-3-carboxylate (intermediate 5d) instead of (1R,5S)- tert -butyl-1-(4-(6-amino-5-(methoxycarbonyl)pyridin-3-yl)phenyl)- 3-azabicyclo[3.1.0]hexane-3-carboxylate (intermediate 1d). (UPLC–MS) t _R 0.92 min; ESI–MS 396 [M+H] ⁺ .

Intermediate 5d : (1S,5R)– tert -butyl–1–(4–(6–amino–5–(methoxycarbonyl)pyridin–3–yl)phenyl)–3–azabicyclo[3.1.0]hexane–3– carboxylate

Racemic tert -butyl-1-(4-(6-amino-5-(methoxycarbonyl)pyridin-3-yl)phenyl)-3-azabicyclo[3.1.0]hexane-3-carboxylate (intermediate 1e, 13) was separated 5 g, 33.0 mmol) using chiral preparative SFC (CHIRALPAK AD–H 5 µm 30 * 250 mm, mobile phase CO ₂ /i–PrOH, 70:30, flow rate 120 g/min; UV detection at 278 nm). The title compound was obtained as an off-white solid after concentration under reduced pressure. Chiral analytical HPLC (CHIRALPAK AD–3 3 µm, 2 * 100 mm, mobile phase CO ₂ /MeOH+0.1% DEA, 60:40, flow rate 1 ml/min; UV detection at 274 nm): t _R 2.90 min, ee=98.7%, [α] _D ²⁰ = −87°.

Example 6 : 2-amino-5-(4-((1R,5S)-3-(1,3-difluoropropan-2-yl)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)- N–((1r,4R)–4–hydroxycyclohexyl)nicotinamide

Into a solution of TFA salt and 5-(4-((1R,5S)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)-2-amino-N-((1r,4R)-4-hydroxycyclohexyl )nicotinamide (intermediate 1a, 70 mg, 0.138 mmol) in acetonitrile (3 ml) was added K ₂ CO ₃ (38.2 mg, 0.276 mmol) and 1,3-difluoropropan-2-yltrifluoromethanesulfonate (intermediate 6a) (37 .8 mg, 0.166 mmol) at room temperature. The reaction mixture was stirred at room temperature. for 1 hour, then diluted with sat. aq. NaHCO ₃ solution and mixed with EtOAc. After phase separation aq. the layer was again extracted with EtOAc. The combined organic layers were washed with brine, dried over MgSO ₄ , filtered and concentrated under reduced pressure. The crude product was purified by prepHPLC (Method 1a) to give the title compound. ^1H NMR (400 MHz, DMSO–d6) δ 8.41 (d, 1H), 8.37 (d, 1H), 8.18 (s, 1H), 7.62 (s, 2H), 7. 35 (d, 1H), 7.25 (m, 3H), 4.64 (d, 3H), 3.74 (m, 4H), 2.89 (d, 4H), 1.87 (t, 5H ), 1.34 (m, 6H), 0.82 (s, 1H). (UPLC–MS) t _R 0.68 min; ESI–MS 471 [M+H] ⁺ .

Intermediate 6a : 1,3-difluoropropane-2-yltrifluoromethanesulfonate

To a solution of 1,3-difluoropropan-2-ol (300 mg, 3.12 mmol) in DCM (8 ml) was added DMAP (26.7 mg, 0.219 mmol) and TEA (0.522 ml, 3.75 mmol) under atmosphere nitrogen at room temperature The reaction mixture was cooled to 0°C and trifluoromethanesulfonic anhydride (0.630 ml, 3.75 mmol) was added. After stirring at 0°C for 60 min and at room temperature. within 3 hours the reaction mixture was diluted with DCM. The organic layer was washed with water, twice aq. solution of citric acid and twice saturated. aq. NaHCO ₃ solution, dried over MgSO ₄ and concentrated at 500 mbar. The title compound was obtained as a crude oil which was used without further purification.

Example 7 : 2-amino-N-(4-hydroxybicyclo[2.2.2]octan-1-yl)-5-(4-((1S,5R)-3-(2-methoxyethyl)-3-azabicyclo[3.1 .0]hexan–1–yl)phenyl)nicotinamide

The title compound was prepared in a manner similar to (1R,5S) -tert -butyl-1-(4-(6-amino-5-(((1r,4R)-4-hydroxycyclohexyl)carbamoyl)pyridin-3-yl )phenyl)-3-azabicyclo[3.1.0]hexane-3-carboxylate (intermediate 1b), except that 2-amino-5-(4-((1S,5R)-3-(2- methoxyethyl)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)nicotinic acid (intermediate 7a) instead of 2-amino-5-(4-((1R,5S)-3-( tert -butoxycarbonyl) –3-azabicyclo[3.1.0]hexan-1-yl)phenyl)nicotinic acid (intermediate 1c) and used 4-aminobicyclo[2.2.2]octan-1-ol hydrochloride instead of trans –4-aminocyclohexanol. The crude product was first purified by prepHPLC (Method 1a) then by reverse phase flash chromatography (Method 3a) to obtain the title compound as a colorless solid. ^1H NMR (400 MHz, DMSO–d6) δ 8.34 (d, 1H), 7.98 (d, 1H), 7.79 (s, 1H), 7.57 (d, 2H), 7. 21 (d, 2H), 6.92 (s, 2H), 4.32 (s, 1H), 3.45 (t, 2H), 3.41–3.29 (m, 2H), 3.27 (s, 3H), 3.08 (d, 1H), 2.68–2.51 (m, 3H), 2.07–2.03 (m, 6H), 1.83–1.79 (m , 1H), 1.65–1.61 (m, 6H), 1.33 (t, 1H), 0.77 (bs, 1H). (UPLC–MS) t _R 0.52 min; ESI–MS 477 [M+H] ⁺ .

Intermediate 7a : 2-amino-5-(4-((1S,5R)-3-(2-methoxyethyl)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)nicotinic acid

The title compound was prepared in a manner similar to that for 2-amino-5-(4-((1R,5S)-3-( tert -butoxycarbonyl)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)nicotinic acid (intermediate 2c), except that methyl-2-amino-5-(4-((1S,5R)-3-(2-methoxyethyl)-3-azabicyclo[3.1.0]hexane-1) was used -yl)phenyl)nicotinate (intermediate 7b) instead of (1R,5S) -tert -butyl-1-(4-(6-amino-5-(methoxycarbonyl)pyridin-3-yl)phenyl)-3-azabicyclo[ 3.1.0]hexane-3-carboxylate (intermediate 1d). (UPLC–MS) t _R 0.32 min; ESI–MS 354 [M+H] ⁺ .

Intermediate 7b : methyl-2-amino-5-(4-((1S,5R)-3-(2-methoxyethyl)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)nicotinate

To a solution of methyl 5-(4-((1S,5R)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)-2-aminonicotinate hydrochloride (intermediate 7c, 210 mg, 0.607 mmol) in acetonitrile (6 ml) K ₂ CO ₃ (168 mg, 0.729 mmol) and 1-bromo-2-methoxyethane (0.068 ml, 0.729 mmol) were added at room temperature. The reaction mixture was stirred at 65°C for 3 hours. The reaction mixture was then diluted with sat. aq. NaHCO ₃ solution and mixed with EtOAc. After phase separation aq. the layer was again extracted with EtOAc. The combined organic layers were washed with brine, dried over MgSO ₄ , filtered and concentrated under reduced pressure to give the title compound, which was used without further purification. (UPLC–MS) t _R 0.62 min; ESI–MS 368 [M+H] ⁺ .

Intermediate 7c : methyl 5-(4-((1S,5R)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)-2-aminonicotinate hydrochloride

Into a solution of (1S,5R) -tert -butyl-1-(4-(6-amino-5-(methoxycarbonyl)pyridin-3-yl)phenyl)-3-azabicyclo[3.1.0]hexane-3-carboxylate ( intermediate 5d, 500 mg, 1.22 mmol) in dioxane (7 ml) was added 4 M HCl in dioxane (1.22 ml, 4.88 mmol) at RT. The reaction mixture was stirred at 60°C for 3 hours and then concentrated under reduced pressure to obtain the title compound as the hydrochloride salt. (UPLC–MS) t _R 0.58 min; ESI–MS 310 [M+H] ⁺ .

Example 8 : 2-amino-5-(2-fluoro-4-(4-isopropylpiperazin-1-yl)phenyl)-N-((1r,4r)-4-hydroxycyclohexyl)nicotinamide

To a solution of 2-amino-5-bromo-N-((1r,4r)-4-hydroxycyclohexyl)nicotinamide (intermediate 3c, 230 mg, 0.731 mmol) in dioxane (4 ml) was added 1-(3-fluoro-4 –(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)-4-isopropylpiperazine (intermediate 8a, 463 mg, 0.731 mmol), 2 N. aq. K ₂ CO ₃ (0.73 ml, 1.46 mmol) and the adduct of PdCl ₂ (dppf) and DCM (29.9 mg, 0.037 mmol) under nitrogen at room temperature. The reaction mixture was stirred at 80°C for 60 min, then diluted with EtOAc and aq. _NaHCO3 . After phase separation aq. the layer was extracted with EtOAc. The organic layers were washed with sat. aq. NaHCO ₃ solution and saline solution, respectively, were dried over MgSO ₄ , filtered and concentrated under reduced pressure. The crude product was purified by prepHPLC (Method 1a) to obtain the title compound as a yellow solid. ^1H NMR (400 MHz, DMSO–d6) δ 8.24–8.16 (m, 2H), 7.95 (d, 1H), 7.39 (t, 1H), 7.06 (s, 2H ), 6.86 (d, 2H), 4.54 (d,1H), 3.69 (m, 1H), 3.41–3.33 (m, 2H), 3.15 (s, 3H) , 2.66 (q, 3H), 1.82 (m, 4H), 1.42–0.88 (m, 10H). (UPLC–MS) t _R 0.48 min; ESI–MS 456 [M+H] ⁺ .

Intermediate 8a : 1-(3-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)-4-isopropylpiperazine

To a solution of 1-(4-bromo-3-fluorophenyl)-4-isopropylpiperazine (intermediate 8b, 212 mg, 0.662 mmol) in dioxane (8 ml) was added bis(pinacolato)diboron (202 mg, 0.794 mmol), KOAc ( 130 mg, 1.323 mmol) and the adduct of PdCl ₂ (dppf) and DCM (27 mg, 0.033 mmol) under nitrogen at room temperature. The reaction mixture was stirred at 90°C for 2 hours, then filtered through a pad of celite and concentrated under reduced pressure to give the title compound, which was used without any further purification. (UPLC–MS) t _R 0.79 min; ESI–MS 349 [M+H] ⁺ .

Intermediate 8b : 1-(4-bromo-3-fluorophenyl)-4-isopropylpiperazine

To a solution of 1-(4-bromo-3-fluorophenyl)piperazine (intermediate 8c, 215 mg, 0.797 mmol) in acetonitrile (7 ml) was added 2-iodopropane (0.095 ml, 0.956 mmol) and K ₂ CO ₃ (220 mg , 1.593 mmol) at room temperature. The reaction mixture was stirred at 65°C for 4 hours, then diluted with EtOAc and aq. _NaHCO3 . After phase separation aq. the layer was extracted with EtOAc. The organic layers were washed with brine, dried over MgSO ₄ , filtered and concentrated under reduced pressure to give the title compound as a colorless oil, which was used without any further purification. (UPLC–MS) t _R 0.66 min; ESI–MS 302 [M+H] ⁺ .

Intermediate 8c : 1-(4-bromo-3-fluorophenyl)piperazine

4 N was added to a solution of tert -butyl-4-(4-bromo-3-fluorophenyl)piperazine-1-carboxylate (300 mg, 0.835 mmol) in dioxane (8 ml). HCl in dioxane (0.84 ml, 3.34 mmol) under nitrogen at room temperature. The reaction mixture was stirred for 2 hours, then diluted with EtOAc and aq. _NaHCO3 . After phase separation aq. the layer was extracted with EtOAc. The organic layers were washed with brine, dried over MgSO ₄ , filtered and concentrated under reduced pressure to give the title compound, which was used without any further purification. (UPLC–MS) t _R 0.60 min; ESI–MS 260 [M+H] ⁺ .

Example 9 : 2-amino-N-((1r,4S)-4-hydroxycyclohexyl)-5-(4-((1S,5R)-3-isopropyl-3-azabicyclo[3.1.0]hexan-1-yl )phenyl)nicotinamide

The title compound was prepared in a manner similar to that for 2-amino-N-((1r,4R)-4-hydroxycyclohexyl)-5-(4-((1R,5S)-3-isopropyl-3-azabicyclo[3.1.0 ]hexan-1-yl)phenyl)nicotinamide (Example 2), except that the TFA salt and 2-amino-5-(4-((1S,5R)-3-isopropyl-3-azabicyclo[3.1. 0]hexan-1-yl)phenyl)nicotinic acid (intermediate 9a) instead of TFA salt and 2-amino-5-(4-((1R,5S)-3-isopropyl-3-azabicyclo[3.1.0]hexane –1-yl)phenyl)nicotinic acid (intermediate 2a). ^1H NMR (400 MHz, DMSO–d6) δ 8.38 (d,1H), 8.31 (d, 1H), 8.11 (d, 1H), 7.58 (d, 2H), 7. 23 (d, 2H), 7.12 (s, 2H), 4.58 (d, 1H), 3.82–3.66 (m, 1H), 3.48–3.34 (m, 3H) , 3.07 (d, 1H), 2.64–2.55 (m, 2H), 1.96–1.77 (m, 5H), 1.50–1.19 (m, 5H), 1 .04 (dd, 6H), 0.75 (dd, 1H). (UPLC–MS) t _R 0.50 min; ESI–MS 435 [M+H] ⁺ .

Intermediate 9a : TFA salt of 2-amino-5-(4-((1S,5R)-3-isopropyl-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)nicotinic acid

The title compound was prepared in a manner similar to that for TFA salt and 2-amino-5-(4-((1R,5S)-3-isopropyl-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)nicotinic acid (intermediate 2a), except that methyl-2-amino-5-(4-((1S,5R)-3-isopropyl-3-azabicyclo[3.1.0]hexan-1-yl)phenyl) was used nicotinate (intermediate 9b) instead of methyl 2-amino-5-(4-((1R,5S)-3-isopropyl-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)nicotinate (intermediate 2b ). (UPLC–MS) t _R 0.37 min; ESI–MS 338 [M+H] ⁺ .

Intermediate 9b : Methyl-2-amino-5-(4-((1R,5S)-3-isopropyl-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)nicotinate

The title compound was prepared in a manner similar to that for methyl 2-amino-5-(4-((1R,5S)-3-isopropyl-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)nicotinate (intermediate compound 2b), except that (1S,5R)-1-(4-bromophenyl)-3-isopropyl-3-azabicyclo[3.1.0]hexane (intermediate 9c) was used instead of (1R,5S)-1 –(4-bromophenyl)-3-isopropyl-3-azabicyclo[3.1.0]hexane (intermediate 2c). (UPLC–MS) t _R 0.62 min; ESI–MS 352 [M+H] ⁺ .

Intermediate 9c : (1S,5R)–1–(4-bromophenyl)–3-isopropyl-3-azabicyclo[3.1.0]hexane

The title compound was prepared in a manner similar to (1R,5S)-1-(4-bromophenyl)-3-isopropyl-3-azabicyclo[3.1.0]hexane (intermediate 2c), except that (1S ,5R)-1-(4-bromophenyl)-3-azabicyclo[3.1.0]hexane (prepared as described in WO 2007/022935) instead of (1R,5S)-1-(4-bromophenyl)-3-azabicyclo[ 3.1.0]hexane. (UPLC–MS) t _R 0.70 min; ESI–MS 280/282 [M+H] ⁺ .

Example 10 : 2-amino-N-((1r,4R)-4-hydroxycyclohexyl)-5-(4-((1R,5S)-3-(2-morpholinoethyl)-3-azabicyclo[3.1.0]hexane –1-yl)phenyl)nicotinamide

The title compound was prepared in a manner similar to that for 2-amino-5-(4-((1R,5S)-3-(1,3-difluoropropan-2-yl)-3-azabicyclo[3.1.0]hexane-1 -yl)phenyl)-N-((1r,4R)-4-hydroxycyclohexyl)nicotinamide (Example 6), except that 4-(2-bromoethyl)morpholine was used instead of 1,3-difluoropropan-2-yltrifluoromethanesulfonate ( intermediate 6a) and the reaction mixture was stirred at 65°C for 1 h. ¹ H NMR (400 MHz, DMSO–d6) δ 8.38 (d, 1H), 8.31 (d, 1H), 8.11 (d, 1H), 7.59 (d, 2H), 7.22 (d, 2H), 7.12 (s, 2H), 4.59 (d, 1H), 3.84–3.67 ( m, 1H), 3.63–3.51 (m, 4H), 3.46–3.39 (m, 2H), 3.09 (d, 1H), 2.71–2.57 (m, 3H), 2.42 (s, 6H), 1.95–1.75 (m, 5H), 1.53–1.15 (m, 5H), 0.77 (d, 1H). (UPLC–MS) t _R 0.49 min; ESI–MS 506 [M+H] ⁺ .

Example 11 : 2-amino-5-(4-((1S,5R)-3-(2-fluoroethyl)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)-N-((1r, 4S)–4-hydroxycyclohexyl)nicotinamide

The title compound was prepared in a manner similar to that for 2-amino-5-(4-((1R,5S)-3-(1,3-difluoropropan-2-yl)-3-azabicyclo[3.1.0]hexane-1 -yl)phenyl)-N-((1r,4R)-4-hydroxycyclohexyl)nicotinamide (Example 5), except that 1-bromo-2-fluoroethane was used instead of 4-(3-bromopropyl)morpholine and the reaction mixture stirred at 60°C for 5 h. ^1H NMR (400 MHz, DMSO–d6) δ 8.38 (d, 1H), 8.31 (d, 1H), 8.11 (d, 1H), 7 .59 (d, 2H), 7.22 (d, 2H), 7.13 (s, 2H), 4.59 (d, 2H), 4.50 (s, 1H), 3.73 (d, 1H), 3.41 (d, 3H), 2.83–2.78 (m, 3H), 1.85 (t, 5H), 1.47–1.16 (m, 6H), 0.78 (s, 1H). (UPLC–MS) t _R 0.47 min; ESI–MS 439 [M+H] ⁺ .

Example 12 : 2-amino-N-((1r,4r)-4-hydroxycyclohexyl)-5-(4-(1-isopropylpyrrolidin-3-yl)phenyl)nicotinamide

The title compound was prepared in a manner similar to that for 2-amino-5-(2-fluoro-4-(4-isopropylpiperazin-1-yl)phenyl)-N-((1r,4r)-4-hydroxycyclohexyl)nicotinamide (example 8), except that 1-isopropyl-3-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)pyrrolidine (intermediate 12a) was used instead 1-(3-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)-4-isopropylpiperazine (intermediate 8a). ^1H NMR (400 MHz, DMSO–d6) δ 8.36 (d,1H), 8.26 (d, 1H), 8.09 (d, 1H), 7.62–7.51 (m, 2H ), 7.37–7.28 (m, 2H), 7.09 (s, 2H), 4.55 (d, 1H), 3.70 (dq, 1H), 3.44–3.34 ( m, 2H), 3.03 (d, 1H), 2.75 (s, 2H), 2.29–2.06 (m, 2H), 1.87–1.68 (m, 5H), 1 .42–1.17 (m, 5H), 1.06 (t, 6H). (UPLC–MS) t _R 0.48 min; ESI–MS 423 [M+H] ⁺ .

Intermediate 12a : 1-isopropyl-3-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)pyrrolidine

The title compound was prepared in a manner similar to 1-(3-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)-4-isopropylpiperazine (intermediate 8a), except that 3-(4-bromophenyl)-1-isopropylpyrrolidine (intermediate 12b) was used instead of 1-(4-bromo-3-fluorophenyl)-4-isopropylpiperazine (intermediate 8b). (UPLC–MS) t _R 0.79 min; ESI–MS 316 [M+H] ⁺ .

Intermediate 12b : 3-(4-bromophenyl)-1-isopropylpyrrolidine

The title compound was prepared in a manner similar to 1-(4-bromo-3-fluorophenyl)-4-isopropylpiperazine (intermediate 8b), except that 3-(4-bromophenyl)pyrrolidine hydrochloride was used instead of 1-(4 –bromo-3-fluorophenyl)piperazine (intermediate 8c). (UPLC–MS) t _R 0.62 min; ESI–MS 269 [M+H] ⁺ .

Example 13 : 2-amino-N-((1r,4S)-4-hydroxycyclohexyl)-5-(4-((1S,5R)-3-(2-morpholinoethyl)-3-azabicyclo[3.1.0]hexane –1-yl)phenyl)nicotinamide

The title compound was prepared in a manner similar to that for 2-amino-5-(4-((1R,5S)-3-(1,3-difluoropropan-2-yl)-3-azabicyclo[3.1.0]hexane-1 -yl)phenyl)-N-((1r,4R)-4-hydroxycyclohexyl)nicotinamide (Example 5), except that 4-(2-bromoethyl)morpholine was used instead of 4-(3-bromopropyl)morpholine. ^1H NMR (400 MHz, DMSO–d6) δ 8.38 (d, 1H), 8.31 (d, 1H), 8.11 (d, 1H), 7.59 (d, 2H), 7. 22 (d, 2H), 7.12 (s, 2H), 4.59 (d, 1H), 3.84–3.67 (m, 1H), 3.63–3.51 (m, 4H) , 3.46–3.39 (m, 2H), 3.09 (d, 1H), 2.71–2.57 (m, 3H), 2.42 (s, 6H), 1.95–1 .75 (m, 5H), 1.53–1.15 (m, 5H), 0.77 (d, 1H). (UPLC–MS) t _R 0.49 min; ESI–MS 506 [M+H] ⁺ .

Example 14 : 2-amino-N-((1r,4R)-4-hydroxy-4-methylcyclohexyl)-5-(4-((1R,5S)-3-(tetrahydro-2H-pyran-4-yl) –3-azabicyclo[3.1.0]hexane-1-yl)phenyl)nicotinamide

The title compound was prepared in a manner similar to (1R,5S) -tert -butyl-1-(4-(6-amino-5-(((1r,4R)-4-hydroxycyclohexyl)carbamoyl)pyridin-3-yl )phenyl)-3-azabicyclo[3.1.0]hexane-3-carboxylate (intermediate 1b), except that 2-amino-5-(4-((1R,5S)-3-(tetrahydro- 2H-pyran-4-yl)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)nicotinic acid (intermediate 14a) instead of 2-amino-5-(4-((1R,5S)-3 -( tert -butoxycarbonyl)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)nicotinic acid (intermediate 1c) and used trans -4-amino-1-methylcyclohexanol instead of trans -4-aminocyclohexanol. The crude product was purified by prepHPLC (Method 1a) to obtain the title compound as the TFA salt. ^1H NMR (400 MHz, DMSO–d6) δ 8.53 (d, 1H), 8.46 (d, 1H), 8.35 (s, 1H), 7.72 (d, 2H), 7. 71 (bs, 2H), 7.41 (d, 2H), 4.06 (dd, 1H), 3.98 (dd, 2H), 3.85–3.43 (m, 5H), 3.27 (t, 2H), 2.26 (dd, 1H), 1.99 (d, 2H), 1.83–1.76 (m, 4H), 1.68–1.56 (m, 3H), 1.51–1.45 (m, 5H), 1.21–1.13 (m, 5H). (UPLC–MS) t _R 0.51 min; ESI–MS 491 [M+H] ⁺ .

Intermediate 14a : 2-amino-5-(4-((1R,5S)-3-(tetrahydro-2H-pyran-4-yl)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl) niacin (TFA salt)

In solution methyl-2-amino-5-(4-((1R,5S)-3-(tetrahydro-2H-pyran-4-yl)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl) nicotinate (intermediate 14b, 1.30 g, 2.97 mmol) in anhydrous THF (16 ml) was added 2 M aq. LiOH solution (4.46 ml, 8.92 mmol) at RT. The reaction mixture was stirred at 65°C for 140 minutes and then concentrated under reduced pressure. The residue was purified by reverse phase chromatography (method 3b). Pure fractions were concentrated and lyophilized to give the title compound as an off-white TFA salt. (UPLC–MS) t _R 0.40 min; ESI–MS 380 [M+H] ⁺ .

Intermediate 14b : methyl-2-amino-5-(4-((1R,5S)-3-(tetrahydro-2H-pyran-4-yl)-3-azabicyclo[3.1.0]hexan-1-yl) phenyl)nicotinate

Into a solution of methyl 5-(4-((1R,5S)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)-2-aminonicotinate hydrochloride (intermediate 14c, 1.18 g, 3.28 mmol) in DCM (16 ml) was added dihydro-2H-pyran-4(3H)-one (0.33 ml, 3.60 mmol) and AcOH (0.38 ml, 6.55 mmol) at RT. The reaction mixture was stirred at 45°C for 30 minutes. Sodium triacetoxyborohydride (1.74 g, 8.19 mmol) was added at room temperature. and the reaction mixture was stirred at 45°C for 1 hour. The reaction mixture was then diluted with sat. NaHCO ₃ solution and mixed with EtOAc. After phase separation aq. the layer was extracted again with EtOAc. The combined organic layers were washed with brine, dried over MgSO ₄ , filtered and concentrated under reduced pressure to give the title compound, which was used without further purification. (UPLC–MS) t _R 0.67 min; ESI–MS 394 [M+H]+.

Intermediate 14c : methyl 5-(4-((1R,5S)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)-2-aminonicotinate hydrochloride

Into a solution of (1R,5S) -tert -butyl-1-(4-(6-amino-5-(methoxycarbonyl)pyridin-3-yl)phenyl)-3-azabicyclo[3.1.0]hexane-3-carboxylate ( intermediate 1d, 1.50 g, 3.66 mmol) in dioxane (25 ml) was added 4 M HCl in dioxane (3.66 ml, 14.7 mmol) at RT. The reaction mixture was stirred at 60°C for 3 hours and then concentrated under reduced pressure to obtain the title compound as the hydrochloride salt. (UPLC–MS) t _R 0.58 min; ESI–MS 310 [M+H] ⁺ .

Example 15 : 2-amino-N-(4-hydroxybicyclo[2.2.2]octan-1-yl)-5-(4-((1R,5S)-3-(2-morpholinoethyl)-3-azabicyclo[3.1 .0]hexan–1–yl)phenyl)nicotinamide

The title compound was prepared in a manner similar to (1R,5S) -tert -butyl-1-(4-(6-amino-5-(((1r,4R)-4-hydroxycyclohexyl)carbamoyl)pyridin-3-yl )phenyl)-3-azabicyclo[3.1.0]hexane-3-carboxylate (intermediate 1b), except that 2-amino-5-(4-((1R,5S)-3-(2- morpholinoethyl)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)nicotinic acid (intermediate 15a) instead of 2-amino-5-(4-((1R,5S)-3-( tert -butoxycarbonyl) –3-azabicyclo[3.1.0]hexan-1-yl)phenyl)nicotinic acid (intermediate 1c) and used 4-aminobicyclo[2.2.2]octan-1-ol hydrochloride instead of trans –4-aminocyclohexanol. The crude product was first purified by prepHPLC (Method 1a) then by reverse phase flash chromatography (Method 3a) to obtain the title compound as a colorless solid. ^1H NMR (400 MHz, DMSO–d6) δ 8.34 (d, 1H), 7.98 (d, 1H), 7.79 (s, 1H), 7.56 (d, 2H), 7. 21 (d, 2H), 6.92 (s, 2H), 4.31 (s, 1H), 3.58–3.56 (m, 4H), 3.09 (d, 1H), 2.64 –2.62 (m, 2H), 2.59–2.41 (m, 9H), 2.07–2.04 (m, 6H), 1.82–1.79 (m, 1H), 1 .65–1.61 (m, 6H), 1.31 (t, 1H), 0.75 (dd, 1H). (UPLC–MS) t _R 0.53 min; ESI–MS 532 [M+H] ⁺ .

Intermediate 15a : 2-amino-5-(4-((1R,5S)-3-(2-morpholinoethyl)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)nicotinic acid

The title compound was prepared in a manner similar to that for 2-amino-5-(4-((1R,5S)-3-( tert -butoxycarbonyl)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)nicotinic acid (intermediate 2c), except that methyl-2-amino-5-(4-((1R,5S)-3-(2-morpholinoethyl)-3-azabicyclo[3.1.0]hexane-1) was used -yl)phenyl)nicotinate (intermediate 15b) instead of (1R,5S) -tert -butyl-1-(4-(6-amino-5-(methoxycarbonyl)pyridin-3-yl)phenyl)-3-azabicyclo[ 3.1.0]hexane-3-carboxylate (intermediate 1d). (UPLC–MS) t _R 0.39 min; ESI–MS 409 [M+H] ⁺ .

Intermediate 15b : Methyl-2-amino-5-(4-((1R,5S)-3-(2-morpholinoethyl)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)nicotinate

The title compound was prepared in a manner similar to that for methyl-2-amino-5-(4-((1S,5R)-3-(2-methoxyethyl)-3-azabicyclo[3.1.0]hexane-1-yl)phenyl )nicotinate (intermediate 7b), except that methyl 5-(4-((1R,5S)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)-2-aminonicotinate hydrochloride ( intermediate 14c) instead of methyl 5-(4-((1S,5R)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)-2-aminonicotinate hydrochloride (intermediate 7c) and used 4-( 2-bromoethyl)morpholine instead of 1-bromo-2-methoxyethane. (UPLC–MS) t _R 0.62 min; ESI–MS 423 [M+H] ⁺ .

Example 16 : 2-amino-N-(3-(hydroxymethyl)bicyclo[1.1.1]pentan-1-yl)-5-(4-((1S,5R)-3-isopropyl-3-azabicyclo[3.1. 0]hexan–1–yl)phenyl)nicotinamide

The title compound was prepared in a manner similar to that for 2-amino-N-((1r,4R)-4-hydroxycyclohexyl)-5-(4-((1R,5S)-3-isopropyl-3-azabicyclo[3.1.0 ]hexan-1-yl)phenyl)nicotinamide (Example 2), except that 2-amino-5-(4-((1S,5R)-3-isopropyl-3-azabicyclo[3.1.0]hexane was used –1-yl)phenyl)nicotinic acid (intermediate 9a) instead of TFA salt and 2-amino-5-(4-((1R,5S)-3-isopropyl-3-azabicyclo[3.1.0]hexane-1- yl)phenyl)nicotinic acid (intermediate 2a) and used (3-aminobicyclo[1.1.1]pentan-1-yl)methanol (intermediate 16a) instead of trans -4-aminocyclohexanol hydrochloride. ^1H NMR (400 MHz, DMSO–d6) δ 9.05 (s, 1H), 8.41 (d,1H), 8.18 (d,1H), 7.61 (d, 2H), 7, 48 (dd, 1H), 7.33–7.17 (m, 4H), 4.55 (t, 1H), 3.51 (d, 3H), 3.31 (s, 2H), 3.08 (s, 2H), 2.07–1.99 (m, 1H), 1.41–1.22 (m, 3H), 1.17–0.98 (m, 9H), 0.79 (d , 1H). (UPLC–MS) t _R 0.54 min; ESI–MS 433 [M+H] ⁺ .

Intermediate 16a : (3-aminobicyclo[1.1.1]pentan-1-yl)methanol

To a solution of HCl salt and methyl-3-aminobicyclo[1.1.1]pentane-1-carboxylate (218 mg, 1.23 mmol) in THF (7 ml), LiAlH ₄ (140 mg, 3.68 mmol) was added in small portions at 0°C. The reaction mixture was stirred at room temperature. for 2 hours and quenched at 0°C with a mixture of THF and water, then filtered through a pad of celite and concentrated under reduced pressure to give the title compound as a yellow oil, which was used without further purification. FIA–MS 114 [M+H] ⁺ .

Example 17 : 2-amino-N-((1r,4S)-4-hydroxycyclohexyl)-5-(4-((1S,5R)-3-(3,3,3-trifluoropropyl)-3-azabicyclo[3.1 .0]hexan–1–yl)phenyl)nicotinamide

The title compound was prepared in a manner similar to that for 2-amino-5-(4-((1R,5S)-3-(1,3-difluoropropan-2-yl)-3-azabicyclo[3.1.0]hexane-1 -yl)phenyl)-N-((1r,4R)-4-hydroxycyclohexyl)nicotinamide (Example 5), except that 3,3,3-trifluoropropyltrifluoromethanesulfonate was used instead of 4-(3-bromopropyl)morpholine and the reaction mixture stirred at room temperature. within 60 min. ^1H NMR (400 MHz, DMSO–d6) δ 8.39 (d, 1H), 8.31 (d, 1H), 8.12 (d, 1H), 7.60 (d, 2H), 7. 23 (d, 2H), 7.12 (s, 2H), 4.59 (d, 1H), 3.79–3.66 (m, 1H), 3.39 (m, 3H), 3.09 (s, 1H), 2.72–2.59 (m, 4H), 1.86 (m, 5H), 1.52–1.18 (m, 6H), 0.79 (s, 1H). (UPLC–MS) t _R 0.59 min; ESI–MS 489 [M+H] ⁺ .

Example 18 : 2-amino-N-((1r,4r)-4-hydroxycyclohexyl)-5-(4-(pyrrolidin-3-yl)phenyl)nicotinamide

Into a solution of tert -butyl-3-(4-(6-amino-5-(((1r,4r)-4-hydroxycyclohexyl)carbamoyl)pyridin-3-yl)phenyl)pyrrolidine-1-carboxylate (intermediate 18a, 120 mg, 0.140 mmol) in dioxane (3 ml) was added 4 N. HCl in dioxane (0.210 ml, 0.839 mmol) under nitrogen at room temperature. and the resulting mixture was stirred for 4 hours. After concentration under reduced pressure, the crude product was purified by reverse phase chromatography (method 3a). Pure fractions were divided between sat. aq. a solution of NaHCO ₃ and EtOAc. The combined organic layers were washed with brine, dried over MgSO ₄ , filtered and concentrated under reduced pressure to give the title compound as a mixture of diastereomers. ^1H NMR (400 MHz, DMSO–d6) δ 8.26 (d,1H), 8.09 (d, 1H), 7.57 (d, 2H), 7.35–7.29 (m, 2H ), 7.09 (s, 2H), 4.55 (d, 1H), 3.77–3.69 (m, 2H), 3.22–3.15 (m, 2H), 3.12– 2.91 (m, 2H), 2.74–2.62 (m, 2H), 2.24–2.07 (m, 1H), 1.83 (dd, 4H), 1.42–1, 16 (m, 5H). (UPLC–MS) t _R 0.42 min; ESI–MS 381 [M+H] ⁺ .

Intermediate 18a : tert -butyl-3-(4-(6-amino-5-(((1r,4r)-4-hydroxycyclohexyl)carbamoyl)pyridin-3-yl)phenyl)pyrrolidine-1-carboxylate

The title compound was prepared in a manner similar to that for 2-amino-5-(2-fluoro-4-(4-isopropylpiperazin-1-yl)phenyl)-N-((1r,4r)-4-hydroxycyclohexyl)nicotinamide (example 8), except that tert -butyl-3-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)pyrrolidine-1-carboxylate (intermediate) was used compound 18b) instead of 1-(3-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)-4-isopropylpiperazine (intermediate 8a). (UPLC–MS) t _R 0.95 min; ESI–MS 481 [M+H] ⁺ .

Intermediate 18b : tert -butyl-3-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)pyrrolidine-1-carboxylate

The title compound was prepared in a manner similar to 1-(3-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)-4-isopropylpiperazine (intermediate 8a), except that tert -butyl-3-(4-bromophenyl)pyrrolidine-1-carboxylate (intermediate 18c) was used instead of 1-(4-bromo-3-fluorophenyl)-4-isopropylpiperazine (intermediate 8b ). (UPLC–MS) t _R 1.43 min; ESI–MS 374 [M+H] ⁺ .

Intermediate 18c : tert -butyl-3-(4-bromophenyl)pyrrolidine-1-carboxylate

To a solution of 3-(4-bromophenyl)pyrrolidine (200 mg, 0.885 mmol) in DCM (7 ml) was added di- tert -butyl dicarbonate (0.308 ml, 1.327 mmol) and TEA (0.247 ml, 1.769 mmol) under a nitrogen atmosphere at .T. The reaction mixture was stirred for 2 hours. The reaction mixture was then diluted with sat. aq. NaHCO ₃ solution and mixed with DCM. After phase separation aq. the layer was again extracted with DCM. The combined organic layers were washed with brine, dried over MgSO ₄ , filtered and concentrated under reduced pressure to give the title compound as a colorless oil which was used without further purification. (UPLC–MS) t _R 1.33 min; ESI–MS 270 [M+H] ⁺ .

Example 19 : 2-amino-N-((1r,4R)-4-hydroxycyclohexyl)-5-(4-((1R,5S)-3-(tetrahydro-2H-pyran-4-yl)-3-azabicyclo [3.1.0]hexan–1–yl)phenyl)nicotinamide

The title compound was prepared in a manner similar to that for 2-amino-N-((1r,4R)-4-hydroxycyclohexyl)-5-(4-((1R,5S)-3-(1-(methylsulfonyl)piperidine-4 –yl)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)nicotinamide (Example 1), except that dihydro-2H-pyran-4(3H)-one was used instead of 1-(methylsulfonyl) piperidine-4-one. ^1H NMR (400 MHz, DMSO–d6) δ 8.39 (s, 1H), 8.31 (d, 1H), 8.12 (d, 1H), 7.62 (dd, 2H), 7. 25 (s, 2H), 7.12 (s, 2H), 4.59 (d, 1H), 3.97–3.63 (m, 4H), 3.40 (m, 2H), 3.31 (m, 2H), 3.19–3.03 (m, 2H), 1.99–1.65 (m, 8H), 1.51–1.16 (m, 8H). (UPLC–MS) t _R 0.48 min; ESI–MS 477 [M+H] ⁺ .

Example 20 : 2-amino-N-((1r,4S)-4-hydroxycyclohexyl)-5-(4-((1S,5R)-3-(2-methoxyethyl)-3-azabicyclo[3.1.0]hexane –1-yl)phenyl)nicotinamide

The title compound was prepared in a manner similar to that for 2-amino-5-(4-((1R,5S)-3-(1,3-difluoropropan-2-yl)-3-azabicyclo[3.1.0]hexane-1 -yl)phenyl)-N-((1r,4R)-4-hydroxycyclohexyl)nicotinamide (Example 5), except that 1-bromo-2-methoxyethane was used instead of 4-(3-bromopropyl)morpholine and the reaction mixture stirred at 60°C for 4 h. ^1H NMR (400 MHz, DMSO–d6) δ 8.38 (s, 1H), 8.11 (s, 1H), 7.59 (d, 2H), 7 .22 (d, 2H), 7.13 (s, 2H), 4.60 (d, 1H), 3.73 (d, 1H), 3.54–3.36 (m, 4H), 3. 27 (s, 3H), 2.64 (m, 4H), 1.87 (m, 6H), 1.40–1.27 (m, 6H), 0.76 (s, 1H). (UPLC–MS) t _R 0.49 min; ESI–MS 451 [M+H] ⁺ .

Example 21 : 2-amino-N-((1R,3S,4R)-3-fluoro-4-hydroxycyclohexyl)-5-(4-((1R,5S)-3-(tetrahydro-2H-pyran-4- yl)-3-azabicyclo[3.1.0]hexane-1-yl)phenyl)nicotinamide

The title compound was prepared in a manner similar to (1R,5S) -tert -butyl-1-(4-(6-amino-5-(((1r,4R)-4-hydroxycyclohexyl)carbamoyl)pyridin-3-yl )phenyl)-3-azabicyclo[3.1.0]hexane-3-carboxylate (intermediate 1b), except that 2-amino-5-(4-((1R,5S)-3-(tetrahydro- 2H-pyran-4-yl)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)nicotinic acid (intermediate 14a) instead of 2-amino-5-(4-((1R,5S)-3 –( tert -butoxycarbonyl)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)nicotinic acid (intermediate 1c) and used (1R,2S,4R)-4-amino-2-fluorocyclohexanol hydrochloride (intermediate compound 21a) instead of trans –4-aminocyclohexanol. The crude product was purified by prepHPLC (Method 1a) to obtain the title compound as the TFA salt. ^1H NMR (400 MHz, DMSO–d6) δ 9.60 (bs, 1H), 8.42 (s, 1H), 8.35 (d, 1H), 8.12 (s, 1H), 7, 64 (bs, 2H), 7.36 (bs, 2H), 7.14 (s, 2H), 4.94 (t, 1H), 4.80 (d, 1H), 4.16–3.82 (m, 4H), 3.81–3.45 (m, 4H), 3.30–3.25 (m, 4H), 2.15 (bs, 1H), 1.88–1.85 (m , 2H), 1.65–1.59 (m, 5H), 1.76–1.27 (m, 3H), 1.18 (bs, 1H). (UPLC–MS) t _R 0.48 min; ESI–MS 495 [M+H] ⁺ .

Intermediate 21a : (1R,2S,4R)–4-amino-2-fluorocyclohexanol hydrochloride

To a solution of benzyl-((1R,3S,4R)-3-fluoro-4-hydroxycyclohexyl)carbamate (intermediate 21b, 376 mg, 1.41 mmol) in EtOH (20 ml) was added palladium on carbon (10%) ( 80 mg, 0.075 mmol) in a nitrogen atmosphere. The reaction vessel was purged with nitrogen three times, then purged with hydrogen three times. After stirring at room temperature. The reaction mixture was passed through a pad of celite for 17 hours and the filter cake was washed with EtOH to obtain a colorless solution. Hydrochloric acid (1.25 M in EtOH, 14 mL, 17.5 mmol) was added slowly and the mixture was stirred at room temperature. for 5 hours. Concentration under reduced pressure gave the title compound as an off-white, hygroscopic solid. FIA ESI–MS 134 [M+H] ⁺ .

Intermediate 21b : benzyl-((1R,3S,4R)-3-fluoro-4-hydroxycyclohexyl)carbamate

To a solution of benzyl-((1R,3S)-3-fluoro-4-oxocyclohexyl)carbamate (intermediate 21c, 768 mg, 2.90 mmol) in MeOH (15 ml) was added NaBH ₄ (274 mg, 7.24 mmol) at 0°C. After stirring for 30 min at 0°C, the reaction mixture was diluted with sat. aq. NH ₄ Cl solution and ensured its heating to room temperature. The solvent was removed under reduced pressure and DCM and water were added. After phase separation aq. the layer was extracted three times with DCM. The combined organic layers were washed with brine, dried over anhydrous MgSO ₄ , filtered and concentrated under reduced pressure to give a white solid. The crude product was purified by normal phase chromatography (method 2b) to obtain the title compound as a colorless solid. (UPLC–MS) t _R 0.76 min; ESI–MS 268 [M+H] ⁺ . The absolute configuration depicted was confirmed by X-ray crystallography.

Intermediate 21c : benzyl-((1R,3S)-3-fluoro-4-oxocyclohexyl)carbamate

To a solution of benzyl-((1R,3S,4S)-3-fluoro-4-hydroxycyclohexyl)carbamate (intermediate 21d, 710 mg, 2.66 mmol) in DCM (26 mL) was added pyridinium chlorochromate (859 mg, 3. 98 mmol). The reaction mixture was stirred for 6 hours at room temperature. Another portion of pyridinium chlorochromate (573 mg, 2.66 mmol) was added and the reaction mixture was stirred for another 18 hours. After addition of DCM and sat. aq. NaHCO ₃ solution, both phases were separated, the aqueous layer was extracted twice with DCM and the combined organic layers were washed with brine, dried over anhydrous MgSO ₄ , filtered and concentrated under reduced pressure. The residue was purified by normal phase chromatography (Method 2b) to give the title compound as a colorless oil. (UPLC–MS) t _R 0.82 min; ESI–MS 266 [M+H] ⁺ .

Intermediate 21d : benzyl-((1R,3S,4S)-3-fluoro-4-hydroxycyclohexyl)carbamate

Benzyl (1R,3R,6S)-7-oxabicyclo[4.1.0]heptan-3-ylcarbamate (intermediate 21e, 1.50 g, 6.07 mmol) was charged to the vessel and TEA trihydrofluoride (4.94 mL) was added , 30.3 mmol). The vessel was sealed and the contents were stirred at 100°C for 2 hours. After cooling, the reaction mixture was slowly poured into a stirred solution of K ₂ CO ₃ (5.87 g, 42.5 mmol) in water (400 ml) and extracted three times with using DCM. The combined organic layers were washed with water and brine, dried over anhydrous MgSO ₄ , filtered and concentrated under reduced pressure. The crude material was purified by normal phase chromatography (method 2b) to obtain a mixture of regioisomers, which was separated by preparative chiral HPLC (column: ChiralPak AD, 20 μm, 50 x 5 cm, flow rate: 90 ml/min, wavelength for detection: 220 nm, mobile phase: n-heptane: EtOH, 90:10 until t=52 min, 85:15 until t=69 min, then 80:20). The title compound was isolated as a colorless oil and characterized by UPLC–MS (SQ13, column: Acquity HSS T3 1.8 μm 2.1 x 50 mm at 60°C, eluent A: water + 0.05% formic acid + 3.75 mM NH ₄ OAc, eluent B: acetonitrile + 0.04% formic acid, gradient: from 5 to 98% B in 1.4 min, flow 1.0 ml/min, t _R 0.81 min; ESI–MS 268 [M+H] ⁺ ).

Intermediate 21e : (1R,3R,6S)–7-oxabicyclo[4.1.0]heptan-3-ylcarbamate

Syn -benzyl-(7-oxabicyclo[4.1.0]heptan-3-yl)carbamate (prepared as described by Gómez-Sánchez et al., Tetrahedron 2005, 61(5), 1207–1219) was separated using preparative chiral HPLC (column: ChiralPak AY, 10 µm, 25x5 cm, flow rate: 30 ml/min, detection wavelength: 214 nm, mobile phase: n-heptane: isopropanol, 80:20). The title compound was isolated as a colorless oil and characterized by chiral HPLC (ChiralPak AY-H, 5 µm, 15 x 0.46 cm, flow rate: 1 ml/min, detection wavelength: 214 nm, mobile phase: hexane:isopropanol, 70:30, t _R 2.24 min, ee 99.2%). (UPLC–MS) t _R 0.90 min; ESI–MS 248 [M+H] ⁺ .

Example 22 : 2-amino-N-((1s,4S)-4-hydroxycyclohexyl)-5-(4-((1R,5S)-3-(tetrahydro-2H-pyran-4-yl)-3-azabicyclo [3.1.0]hexan–1–yl)phenyl)nicotinamide

The title compound was prepared in a manner similar to (1R,5S) -tert -butyl-1-(4-(6-amino-5-(((1r,4R)-4-hydroxycyclohexyl)carbamoyl)pyridin-3-yl )phenyl)-3-azabicyclo[3.1.0]hexane-3-carboxylate (intermediate 1b), except that 2-amino-5-(4-((1R,5S)-3-(tetrahydro- 2H-pyran-4-yl)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)nicotinic acid (intermediate 14a) instead of 2-amino-5-(4-((1R,5S)-3 -( tert -butoxycarbonyl)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)nicotinic acid (intermediate 1c) and used cis -4-aminocyclohexanol instead of trans -4-aminocyclohexanol. The crude product was purified by prepHPLC (Method 1a) to obtain the title compound as the TFA salt. ^1H NMR (400 MHz, DMSO–d6) δ 9.50 (bs, 1H), 8.41–8.33 (m, 2H), 8.17 (d, 1H), 7.62 (bs, 2H ), 7.26 (bs, 2H), 7.14 (s, 1H), 4.42 (d, 1H), 4.05–3.79 (m, 5H), 3.34–3.24 ( m, 4H), 3.11 (bs, 1H), 2.70–2.50 (m, 1H), 1.83–1.71 (m, 7H), 1.62–1.47 (m, 7H), 1.35–1.29 (m, 1H). (UPLC–MS) t _R 0.52 min; ESI–MS 477 [M+H] ⁺ .

Example 23 : 5-(4-(2-azaspiro[3.3]heptan-2-ylmethyl)phenyl)-2-amino-N-((1r,4r)-4-hydroxycyclohexyl)nicotinamide

The title compound was prepared in a manner similar to that for 2-amino-5-(2-fluoro-4-(4-isopropylpiperazin-1-yl)phenyl)-N-((1r,4r)-4-hydroxycyclohexyl)nicotinamide (example 8), except that 2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzyl)-2-azaspiro[3.3]heptane (intermediate) was used 23a) instead of 1-(3-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)-4-isopropylpiperazine (intermediate 8a). ^1H NMR (400 MHz, DMSO–d6) δ 8.40 (d, 1H), 8.30 (d, 1H), 8.14 (d, 1H), 7.61 (d, 2H), 7. 33 (d, 2H), 7.14 (s, 2H), 4.58 (d, 1H), 3.79–3.66 (m, 1H), 3.55 (s, 2H), 3.41 (dt, 1H), 3.14 (s, 3H), 2.07 (t, 4H), 1.94–1.72 (m, 6H), 1.47–1.20 (m, 5H) ( UPLC–MS) t _R 0.51 min; ESI–MS 421 [M+H] ⁺ .

Intermediate 23a : 2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzyl)-2-azaspiro[3.3]heptane

2-Azaspiro[ 3.3]heptane (148 mg, 1.11 mmol) and cesium carbonate (428 mg, 1.31 mmol) under nitrogen at room temperature. The reaction mixture was stirred for 60 minutes, then diluted with sat. aq. a solution of NaHCO ₃ and EtOAc. After phase separation aq. the layer was extracted with EtOAc and the combined organic layers were washed with brine, dried over MgSO ₄ , filtered and concentrated under reduced pressure to give the title compound, which was used without further purification. (UPLC–MS) t _R 0.82 min; ESI–MS 314 [M+H] ⁺ .

Example 24 : 2-amino-N-((1r,4R)-4-hydroxy-4-methylcyclohexyl)-5-(4-((1R,5S)-3-isopropyl-3-azabicyclo[3.1.0]hexane –1-yl)phenyl)nicotinamide

The title compound was prepared in a manner similar to that for 2-amino-N-((1r,4R)-4-hydroxycyclohexyl)-5-(4-((1R,5S)-3-isopropyl-3-azabicyclo[3.1.0 ]hexan-1-yl)phenyl)nicotinamide (Example 2), except that trans -4-amino-1-methylcyclohexanol was used instead of trans -4-aminocyclohexanol hydrochloride. ^1H NMR (400 MHz, DMSO–d6) δ 8.43 (d, 1H), 8.34 (d, 1H), 8.16 (d, 1H), 7.68 (d, 2H), 7. 39 (d, 2H), 4.06 (dd, 1H), 3.86–3.41 (m, 9H), 2.26 (dt, 1H), 1.78 (d, 2H), 1.68 –1.55 (m, 2H), 1.47 (t, 4H), 1.33 (dd, 6H), 1.18 (s, 4H). (UPLC–MS) t _R 0.53 min; ESI–MS 449 [M+H] ⁺ .

Example 25 : 2-amino-5-(4-((1R,5S)-3-(4,4-difluorocyclohexyl)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)-N-(( 1r,4R)–4–hydroxycyclohexyl)nicotinamide

The title compound was prepared in a manner similar to that for 2-amino-N-((1r,4R)-4-hydroxycyclohexyl)-5-(4-((1R,5S)-3-(1-(methylsulfonyl)piperidine-4 –yl)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)nicotinamide (Example 1), except that 4,4-difluorocyclohexanone was used instead of 1-(methylsulfonyl)piperidin-4-one. ^1H NMR (400 MHz, DMSO–d6) δ 8.38 (d, 1H), 8.32 (d, 1H), 8.11 (d, 1H), 7.59 (d, 2H), 7. 23 (d, 2H), 7.13 (s, 2H), 4.59 (d, 1H), 3.73 (d, 1H), 3.39 (d, 2H), 3.09 (d, 1H ), 2.68–2.55 (m, 2H), 2.37 (d, 2H), 2.00 (s, 2H), 1.84 (d, 8H), 1.64 (d, 2H) , 1.46–1.16 (m, 5H), 0.78 (s, 1H). (UPLC–MS) t _R 0.56 min; ESI–MS 511 [M+H] ⁺ .

Example 26 : 2-amino-N-((1r,4S)-4-hydroxycyclohexyl)-5-(4-((1S,5R)-3-(oxetan-3-yl)-3-azabicyclo[3.1.0 ]hexan–1–yl)phenyl)nicotinamide

Into a solution of TFA salt and 5-(4-((1S,5R)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)-2-amino-N-((1r,4S)-4-hydroxycyclohexyl )nicotinamide (intermediate 5a, 70 mg, 0.138 mmol) in DCM (4 ml) was added oxetan-3-one (12.0 mg, 0.166 mmol) and AcOH (0.012 ml, 0.207 mmol) under nitrogen at room temperature . The reaction mixture was stirred at 45°C for 30 minutes. Sodium triacetoxyborohydride (58.6 mg, 0.276 mmol) was added at room temperature. and the reaction mixture was stirred at 45°C for 3 hours. The reaction mixture was then diluted with sat. aq. NaHCO ₃ solution and mixed with EtOAc. After phase separation aq. the layer was again extracted with EtOAc. The combined organic layers were washed with brine, dried over MgSO ₄ , filtered and concentrated under reduced pressure. The crude product was first purified by prepHPLC (Method 1a) then by reverse phase flash chromatography (Method 3a) to obtain the title compound as a colorless solid. (UPLC–MS) t _R 0.48 min; ESI–MS 449 [M+H] ⁺ .

Example 27 : 2-amino-5-(4-((1S,5R)-3-(4,4-difluorocyclohexyl)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)-N-(( 1r,4S)–4–hydroxycyclohexyl)nicotinamide

The title compound was prepared in a manner similar to that for 2-amino-N-((1r,4S)-4-hydroxycyclohexyl)-5-(4-((1S,5R)-3-(oxetan-3-yl)-3 -azabicyclo[3.1.0]hexan-1-yl)phenyl)nicotinamide (Example 26), except that 4,4-difluorocyclohexanone was used instead of oxetan-3-one. After purification, the title compound was obtained as the TFA salt. ^1H NMR (400 MHz, DMSO–d6) δ 9.50 (bs, 1H), 8.38 (d, 1H), 8.32 (d, 1H), 8.11 (d, 1H), 7, 59 (d, 2H), 7.23 (d, 2H), 7.13 (s, 2H), 4.59 (d, 1H), 4.12 (bs, 1H), 3.73 (d, 1H ), 3.39 (d, 2H), 3.09 (d, 1H), 2.68–2.55 (m, 2H), 2.37 (d, 2H), 2.00 (s, 2H) , 1.84 (d, 8H), 1.64 (d, 2H), 1.46–1.16 (m, 5H). (UPLC–MS) t _R 0.56 min; ESI–MS 511 [M+H] ⁺ .

Example 28 : 2-amino-N-(3-(hydroxymethyl)bicyclo[1.1.1]pentan-1-yl)-5-(4-((1S,5R)-3-(tetrahydro-2H-pyran-4 –yl)-3-azabicyclo[3.1.0]hexane-1-yl)phenyl)nicotinamide

The title compound was prepared in a manner similar to (1R,5S) -tert -butyl-1-(4-(6-amino-5-(((1r,4R)-4-hydroxycyclohexyl)carbamoyl)pyridin-3-yl )phenyl)-3-azabicyclo[3.1.0]hexane-3-carboxylate (intermediate 1b), except that 2-amino-5-(4-((1S,5R)-3-(tetrahydro- 2H-pyran-4-yl)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)nicotinic acid (intermediate 28a) instead of 2-amino-5-(4-((1R,5S)-3 –( tert -butoxycarbonyl)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)nicotinic acid (intermediate 1c) and (3-aminobicyclo[1.1.1]pentan-1-yl)methanol (intermediate compound 16a) instead of trans –4-aminocyclohexanol. The crude product was first purified by prepHPLC (Method 1a) then by reverse phase flash chromatography (Method 3a) to obtain the title compound as a colorless solid. ^1H NMR (400 MHz, DMSO–d6) δ 9.35 (bs, 1H), 9.06 (s, 1H), 8.44 (s, 1H), 8.19 (s, 1H), 7, 68 (d, 2H), 7.37 (d, 2H), 7.26 (bs, 2H), 4.57 (t, 1H), 4.12 (d, 1H), 3.97 (bs, 1H ), 3.80–3.56 (m, 2H), 3.51 (d, 2H), 3.35–3.20 (m, 6H), 2.29–2.23 (m, 1H), 2.11–2.05 (m, 1H), 1.99 (bs, 1H), 1.97 (s, 6H), 1.67–1.61 (m, 1H), 1.19–1, 14 (bs, 1H). (UPLC–MS) t _R 0.50 min; ESI–MS 475 [M+H] ⁺ .

Intermediate 28a : 2-amino-5-(4-((1S,5R)-3-(tetrahydro-2H-pyran-4-yl)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl) a nicotinic acid

The title compound was prepared in a manner similar to that for 2-amino-5-(4-((1R,5S)-3-( tert -butoxycarbonyl)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)nicotinic acid (intermediate 2c), except that methyl-2-amino-5-(4-((1S,5R)-3-(tetrahydro-2H-pyran-4-yl)-3-azabicyclo[3.1 .0]hexan-1-yl)phenyl)nicotinate (intermediate 28b) instead of (1R,5S)-tert-butyl-1-(4-(6-amino-5-(methoxycarbonyl)pyridin-3-yl)phenyl )-3-azabicyclo[3.1.0]hexane-3-carboxylate (intermediate 1d). (UPLC–MS) t _R 0.37 min; ESI–MS 380 [M+H] ⁺ .

Intermediate 28b : methyl-2-amino-5-(4-((1S,5R)-3-(tetrahydro-2H-pyran-4-yl)-3-azabicyclo[3.1.0]hexan-1-yl) phenyl)nicotinate

The title compound was prepared in a manner similar to that for methyl-2-amino-5-(4-((1R,5S)-3-(tetrahydro-2H-pyran-4-yl)-3-azabicyclo[3.1.0]hexane -1-yl)phenyl)nicotinate (intermediate 14b), except that methyl 5-(4-((1S,5R)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl) was used –2-aminonicotinate hydrochloride (intermediate 7c) instead of methyl 5-(4-((1R,5S)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)-2-aminonicotinate hydrochloride (intermediate 14c ). (UPLC–MS) t _R 0.62 min; ESI–MS 394 [M+H] ⁺ .

Example 29 : 2-amino-5-(2-fluoro-4-(piperazin-1-yl)phenyl)-N-((1r,4r)-4-hydroxycyclohexyl)nicotinamide

The title compound was prepared in a manner similar to that for 2-amino-N-((1r,4r)-4-hydroxycyclohexyl)-5-(4-(pyrrolidin-3-yl)phenyl)nicotinamide (Example 18), except that they used tert -butyl-4-(4-(6-amino-5-(((1r,4r)-4-hydroxycyclohexyl)carbamoyl)pyridin-3-yl)-3-fluorophenyl)piperazine-1-carboxylate ( intermediate 29a) instead of 3-(4-(6-amino-5-(((1r,4r)-4-hydroxycyclohexyl)carbamoyl)pyridin-3-yl)phenyl)pyrrolidine-1-carboxylate (intermediate 18a). ^1H NMR (400 MHz, DMSO–d6) δ 8.29–8.16 (m, 2H), 7.95 (d, 1H), 7.38 (t, 1H), 7.06 (s, 2H ), 6.88–6.75 (m, 2H), 4.54 (d, 1H), 3.69 (m, 1H), 3.42–3.32 (m, 2H), 3.17 ( dd, 4H), 2.92 (dt, 4H), 1.88–1.73 (m, 4H), 1.43–1.15 (m, 4H). (UPLC–MS) t _R 0.42 min; ESI–MS 414 [M+H] ⁺ .

Intermediate 29a : tert -butyl-4-(4-(6-amino-5-(((1r,4r)-4-hydroxycyclohexyl)carbamoyl)pyridin-3-yl)-3-fluorophenyl)piperazine-1-carboxylate

The title compound was prepared in a manner similar to that for 1-(3-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)-4-isopropylpiperazine (Example 8 ), except that tert -butyl-4-(3-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)piperazine-1- was used carboxylate (intermediate 29b) instead of 1-(3-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)-4-isopropylpiperazine (intermediate 8a) . The reaction mixture was stirred at 80°C for 60 minutes. and purified by normal phase chromatography (Method 2b) to obtain the title compound as a brownish solid. (UPLC–MS) t _R 0.96 min; ESI–MS 514 [M+H] ⁺ .

Intermediate 29b : tert -butyl-4-(3-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)piperazine-1-carboxylate

The title compound was prepared in a manner similar to tert -butyl-4-(3-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)piperazine-1 -carboxylate (intermediate 8a), except that tert -butyl-4-(4-bromo-3-fluorophenyl)piperazine-1-carboxylate was used instead of 1-(4-bromo-3-fluorophenyl)piperazine (intermediate 8b). (UPLC–MS) t _R 1.38 min; ESI–MS 407 [M+H] ⁺ .

Example 30 : 2-amino-N-(3-(hydroxymethyl)bicyclo[1.1.1]pentan-1-yl)-5-(4-((1R,5S)-3-(tetrahydro-2H-pyran-4 –yl)-3-azabicyclo[3.1.0]hexane-1-yl)phenyl)nicotinamide

The title compound was prepared in a manner similar to (1R,5S) -tert -butyl-1-(4-(6-amino-5-(((1r,4R)-4-hydroxycyclohexyl)carbamoyl)pyridin-3-yl )phenyl)-3-azabicyclo[3.1.0]hexane-3-carboxylate (intermediate 1b), except that 2-amino-5-(4-((1R,5S)-3-(tetrahydro- 2H-pyran-4-yl)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)nicotinic acid (intermediate 14a) instead of 2-amino-5-(4-((1R,5S)-3 –( tert -butoxycarbonyl)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)nicotinic acid (intermediate 1c) and (3-aminobicyclo[1.1.1]pentan-1-yl)methanol (intermediate compound 16a) instead of trans –4-aminocyclohexanol. The crude product was first purified by prepHPLC (Method 1a) then by reverse phase flash chromatography (Method 3a) to obtain the title compound as a colorless solid. ^1H NMR (400 MHz, DMSO–d6) δ 9.05 (s, 1H), 8.41 (d,1H), 8.18 (d,1H), 7.61 (d, 2H), 7, 24–7.22 (m, 4H), 4.55 (t, 1H), 3.87–3.84 (m, 2H), 3.51 (d, 2H), 3.48–3.25 ( m, 2H), 3.10 (bs, 1H), 2.56–2.47 (m, 4H), 1.97 (s, 6H), 1.96–1.74 (m, 3H), 1 .39–0.30 (m, 3H), 0.77 (bs, 1H). (UPLC–MS) t _R 0.51 min; ESI–MS 475 [M+H] ⁺ .

Example 31 : 2-amino-N-((1r,4R)-4-hydroxycyclohexyl)-5-(4-(((R)-2-methylpyrrolidin-1-yl)methyl)phenyl)nicotinamide

The title compound was prepared in a manner similar to that for 2-amino-5-(2-fluoro-4-(4-isopropylpiperazin-1-yl)phenyl)-N-((1r,4r)-4-hydroxycyclohexyl)nicotinamide (example 8), except that (R)-(4-((2-methylpyrrolidin-1-yl)methyl)phenyl)boronic acid (intermediate 31a) was used instead of 1-(3-fluoro-4-(4, 4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)-4-isopropylpiperazine (intermediate 8a). Purification was carried out using normal phase chromatography (method 2b) to obtain the title compound as a yellow solid. ^1H NMR (400 MHz, DMSO–d6) δ 8.75 (d, 1H), 8.65 (d, 1H), 8.48 (d, 1H), 7.81 (dd, 2H), 7. 63 (d, 2H), 4.52 (dd, 1H), 4.18 (dd, 1H), 3.69 (ddt, 1H), 3.51–3.31 (m, 2H), 3.25 –3.04 (m, 3H), 2.19 (ddt, 1H), 1.82 (td, 6H), 1.64–1.52 (m, 1H), 1.39–1.15 (m , 7H). (UPLC–MS) t _R 0.44 min; ESI–MS 409 [M+H] ⁺ .

Intermediate 31a : (R)–(4–((2-methylpyrrolidin-1-yl)methyl)phenyl)boronic acid

To a solution of (4-(bromomethyl)phenyl)boronic acid (125 mg, 0.582 mmol) in acetonitrile (4 ml) was added K ₂ CO ₃ (161 mg, 1.164 mmol) and (R)-2-methylpyrrolidine (54.5 mg , 0.640 mmol) in a nitrogen atmosphere at room temperature. The reaction mixture was stirred for 60 minutes, then filtered through a pad of celite. The filtrate was concentrated under reduced pressure to give the title compound as an off-white solid, which was used without further purification. (UPLC–MS) t _R 0.33 min; ESI–MS 220 [M+H] ⁺ .

Example 32 : 2-amino-N-((1r,4S)-4-hydroxy-4-methylcyclohexyl)-5-(4-((1S,5R)-3-isopropyl-3-azabicyclo[3.1.0]hexane –1-yl)phenyl)nicotinamide

The title compound was prepared in a manner similar to that for 2-amino-N-((1r,4R)-4-hydroxycyclohexyl)-5-(4-((1R,5S)-3-isopropyl-3-azabicyclo[3.1.0 ]hexan-1-yl)phenyl)nicotinamide (Example 2), except that the TFA salt and 2-amino-5-(4-((1S,5R)-3-isopropyl-3-azabicyclo[3.1. 0]hexan-1-yl)phenyl)nicotinic acid (intermediate 9a) instead of TFA salt and 2-amino-5-(4-((1R,5S)-3-isopropyl-3-azabicyclo[3.1.0]hexane -1-yl)phenyl)nicotinic acid (intermediate 2a) and used trans -4-amino-1-methylcyclohexanol instead of trans -4-aminocyclohexanol hydrochloride. ^1H NMR (400 MHz, DMSO–d6) δ 8.43 (d, 1H), 8.34 (d, 1H), 8.16 (d, 1H), 7.68 (d, 2H), 7. 39 (d, 2H), 4.06 (dd, 1H), 3.86–3.41 (m, 9H), 2.26 (dt, 1H), 1.78 (d, 2H), 1.68 –1.55 (m, 2H), 1.47 (t, 4H), 1.33 (dd, 6H), 1.18 (s, 4H). (UPLC–MS) t _R 0.50 min; ESI–MS 449 [M+H] ⁺ .

Example 33 : 2-amino-5-(4-((1S,5R)-3-(4,4-difluorocyclohexyl)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)-N-(( 1r,4S)–4–hydroxycyclohexyl)nicotinamide

The title compound was prepared in a manner similar to that for 2-amino-N-((1r,4S)-4-hydroxycyclohexyl)-5-(4-((1S,5R)-3-(oxetan-3-yl)-3 -azabicyclo[3.1.0]hexan-1-yl)phenyl)nicotinamide (Example 26), except that dihydro-2H-pyran-4(3H)-one was used instead of oxetan-3-one. ^1H NMR (400 MHz, DMSO–d6) δ 8.39 (s, 1H), 8.31 (d, 1H), 8.12 (d, 1H), 7.62 (dd, 2H), 7. 25 (s, 2H), 7.12 (s, 2H), 4.59 (d, 1H), 3.97–3.63 (m, 4H), 3.40 (m, 2H), 3.31 (m, 2H), 3.19–3.03 (m, 2H), 1.99–1.65 (m, 8H), 1.51–1.16 (m, 8H). (UPLC–MS) t _R 0.47 min; ESI–MS 477 [M+H] ⁺ .

Example 34 : 2-amino-N-(4-hydroxybicyclo[2.2.2]octan-1-yl)-5-(4-((1R,5S)-3-(tetrahydro-2H-pyran-4-yl) –3-azabicyclo[3.1.0]hexan-1-yl)phenyl)nicotinamide (also referred to as Compound A herein)

Into a solution of TFA salt and 2-amino-5-(4-((1R,5S)-3-(tetrahydro-2H-pyran-4-yl)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl )nicotinic acid (intermediate 14a, 4.10 g, 8.14 mmol) and 4-aminobicyclo[2.2.2]octan-1-ol hydrochloride (2.17 g, 12.2 mmol) in anhydrous DMF (60 ml ) N-methylmorpholine (2.24 ml, 20.4 mmol) and HATU (4.64 g, 12.2 mmol) were added under nitrogen at room temperature. The reaction mixture was stirred for 2 hours and then diluted with sat. aq. NaHCO ₃ solution and extracted three times with EtOAc. The combined organic extracts were washed with brine, dried over MgSO ₄ , filtered and concentrated under reduced pressure. The crude product was purified by reverse phase chromatography (method 3b). Pure fractions were treated with sat. aq. NaHCO ₃ solution and extracted three times with EtOAc. The combined organic extracts were washed with brine, dried over MgSO ₄ , filtered and concentrated under reduced pressure to give the title compound as an off-white solid. The absolute configuration depicted was confirmed by X-ray crystallography of the title compound in complex with the ALK-2-kinase domain. ^1H NMR (400 MHz, DMSO–d6) δ 8.34 (d, 1H), 7.98 (d, 1H), 7.79 (s, 1H), 7.57 (d, 2H), 7. 23 (d, 2H), 6.92 (s, 2H), 4.31 (s, 1H), 3.91–3.78 (m, 2H), 3.40 (bs, 1H), 3.33 –3.24 (m, 2H), 3.11 (d, 1H), 2.57 (bs, 1H), 2.50–2.34 (m, 1H), 2.34 (bs, 1H), 2.12–1.94 (m, 6H), 1.90–1.72 (m, 3H), 1.71–1.51 (m, 6H), 1.51–1.34 (m, 2H ), 1.31 (t, 1H), 0.82–0.68 (m, 1H). (UPLC–MS) t _R 0.54 min; ESI–MS 503 [M+H] ⁺ . Chiral HPLC (ChiralPak Id, 5 µm, flow rate: 1 ml/min, detection wavelength: 270 nm, mobile phase: heptane:isopropanol, 60:40 (+ 0.1% diethylamine)): t _R 18.7 min, ee 92.3%.

Alternate Example 34A. Into a solution of 2-amino-5-(4-((1R,5S)-3-(tetrahydro-2H-pyran-4-yl)-3-azabicyclo[3.1.0]hexan-1-yl) phenyl)nicotinic acid hydrochloride (1 kg, 1.683 mol) and 4-aminobicyclo[2.2.2]octan-1-ol hydrochloride (343.9 g, 1.935 mol) in DMF (3500 ml) was added Et ₃ N (681.2 g, 6.732 mol ) and HATU (767.9 g, 2.019 mol) at RT. The reaction mixture was stirred at room temperature. for 1 hour. The mixture was heated to IT=45°C, a 5% solution of NH ₃ ⋅H ₂ O (5200 g) was added. Stir for approximately 30 minutes and add additional 5% NH ₃ ⋅H ₂ O solution (1800 g). The mixture was heated to IT=45°C for 2 hours. The mixture was cooled to IT=22°C, filtered, and the wet filter cake was washed with H ₂ O (1500 ml x 3). The wet filter cake was dried under vacuum at 45°C for 24 hours. The crude product was dissolved in acetone (3000 ml), then filtered to remove some undissolved solid. The filtrate was heated to IT=50°C. H ₂ O (2000 ml) was added. The mixture was stirred at IT=50°C for 30 minutes until a white precipitate formed. H ₂ O (4000 ml) was added slowly. The mixture was stirred at IT=50°C for 2 hours. The mixture was cooled to IT=22°C for 2 hours, filtered, the wet filter cake was washed with acetone: H ₂ O=1:2 (v/v, 1000 ml × 2). The wet filter cake was dried under vacuum at 45°C for 24 hours. A total of 760 g of a white solid was obtained (yield 89%, ee 99.4%).

^1H NMR (DMSO–d6) δ: 8.32 (d, J=2.3 Hz, 1H), 7.97 (d, J=2.3 Hz, 1H), 7.77 (s, 1H) , 7.54 (d, J=8.3 Hz, 2H), 7.20 (d, J=8.4 Hz, 2H), 6.90 (s, 2H), 4.31 (s, 1H) , 3.82 (m, 2H), 3.29 (m, 2H), 3.07 (d, J=8.5 Hz, 1H), 2.54 (d, J=8.3 Hz, 1H) , 2.44 (dd, J=8.5, 3.5 Hz, 1H), 2.37 (m, 1H), 2.31 (td, J=10.2, 5.0 Hz, 1H), 2.04 (m, 6H), 1.80 (dt, J=7.9, 3.8 Hz, 1H), 1.71 (d, J=12.3 Hz, 1H), 1.65 (d , J=11.5 Hz, 1H), 1.62 (m, 6H), 1.38 (m, 1H), 1.34 (m, 1H), 1.29 (t, J=3.9 Hz , 1H), 0.73 (dd, J=7.9, 3.6 Hz, 1H).

¹³ C NMR (DMSO–d6) δ: 167.80, 157.69, 148.28, 141.27, 134.91, 134.79, 126.40, 125.66, 123.53, 111.01, 66.22, 65.59, 59.10, 55.46, 52.04, 33.72, 31.92, 31.77, 30.59, 29.61, 24.14, 17.20.

MS(ESI–TOF): 503.3018 [M+H]+.

The starting material (hydrochloride salt) was prepared as follows.

Methyl-2-amino-5-(4-((1R,5S)-3-(tetrahydro-2H-pyran-4-yl)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)nicotinate was suspended dihydrochloride (10 g, 19.5 mmol, 1.0 eq) in MeOH (31.7 g). A solution of NaOH (2.9 g, 72.2 mmol, 3.7 eq) in H ₂ O (10 g) was then added. The reaction mixture was heated to 45 ± 5°C and stirred for more than 3 hours to obtain a suspension.

5–6 N was added to another bottle containing acetone (200 g). HCl in i –PrOH (14.8 g, 97.6 mmol, 5 eq.). The solution was heated to 47 ± 3°C. The above MeOH suspension was then added dropwise to the mixture and stirred at 47 ± 3°C for 3 hours. The mixture was cooled to 23 ± 3°C and stirred for 3 hours. After filtration, the wet filter cake was washed with acetone (40 g). The wet filter cake was dried under vacuum at 55°C for 8 hours. 2-amino-5-(4-((1 R ,5 S )-3-(tetrahydro-2 H -pyran-4-yl)- 3-Azabicyclo[3.1.0]hexan-1-yl)phenyl)nicotinic acid hydrochloride (12.3 g, 99.3% purity by HPLC, 62.1% analytical yield) as an off-white solid .

^1H NMR (DMSO–d6) δ: 11.53 (br s, 1H), 8.64 (br s, 1H), 8.54 (br s, 1H), 7.72–8.42 (m, 2H), 7.64 (br d, J=7.9 Hz, 2H), 7.38 (br d, J=7.8 Hz, 2H), 3.85–4.04 (m, 3H), 3.40–3.73 (m, 4H), 3.15–3.33 (m, 2H), 2.18 (br d, J=3.9 Hz, 1H), 1.95–2.12 (m, 4H), 1.88 (br d, J=10.0 Hz, 1H), 1.05 (br t, J=6.4 Hz, 1H).

¹³ C NMR (DMSO–d6) δ: 167.2, 155.8, 144.3, 142.0, 139.6, 133.8, 127.7, 126.3, 124.1, 110.0, 65.8, 62.5, 55.7, 53.2, 29.9, 28.8, 28.7, 23.5, 16.6.

MS(ESI–TOF): 380.1974 [M+H]+.

Starting material, 2-amino-5-(4-(( 1R , 5S )-3-(tetrahydro- 2H -pyran-4-yl)-3-azabicyclo[3.1.0]hexan-1-yl) Phenyl)nicotinate dihydrochloride was prepared as follows.

1 R ,5 S )-1-(4-bromophenyl)-3-(tetrahydro-2 H -pyran-4-yl)-3-azabicyclo[3.1.0]hexane-3-ium was loaded into a 500 ml round bottom flask chloride (20 g, 1 eq.), EA (200 ml) and 25% K ₂ CO ₃ (62 g). The mixture was stirred for 30 minutes until all solids had dissolved. After phase separation, the organic layer was concentrated. 2-methyl-2-butanol (48 g, 60 ml) was added. The organic layer was concentrated. 2-methyl-2-butanol (144 g, 180 ml) was added.

The mixture was transferred into a 500 ml Redlay. Add K ₂ CO ₃ (18.8 g, 2.5 eq.) and methyl 2-amino-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)nicotinate (16.8 g, 1.04 eq; see intermediate 1e). The mixture was degassed three times with N ₂ . The mixture was heated to IT=50 ± 5°C for 1 hour. Pd(dppf)Cl ₂ (1.2 g, 0.03 eq.) was added. The mixture was heated to IT=70 ± 3°C and stirred for 2 hours. After cooling to 22°C, H ₂ O (120 g) and EA (180 g) were added and stirred for 30 minutes. MCC (6 g) was added and the mixture was filtered through MCC. The filter cake was washed with EA (54 g). After phase separation, the organic layer was washed with 5% NaCl (124 g). Quadrasil MP (heavy metal scavenger from Johnson Matthey, 6 g) was then added to the organic layer. The mixture was heated to IT=55°C for 8 hours, filtered through MCC and washed with EA (54 g). Quadrasil MP (2 g) was added to the organic layer. The mixture was heated to IT=55°C for 6 hours, filtered through CMC and washed with EA (54 g). The organic layer was concentrated. Acetone (158 g, 200 ml) was added. After stirring at IT=22 ± 3°C for 30 min, the mixture was heated to IT=40 ± 3°C. 15.5% HCl (38.4 g) was added dropwise at IT <50°C. The mixture was stirred at IT=45 ± 3°C for 1 hour. The mixture was cooled to 22 ± 3°C. The mixture was stirred at 22 ± 3°C for 1 hour and filtered. The filter cake was washed with acetone (32 g × 2). The wet filter cake was dried under vacuum at 50°C for at least 8 hours. The starting material was 22.5 g of a white solid (purity 97.1% by HPLC, water content 5.2%, yield at analysis 87%).

^1H NMR (DMSO–d6) δ: 11.43 (br d, J=5.7 Hz, 1H), 8.62–8.80 (m, 2H), 7.85–8.58 (m, 2H), 7.67 (d, J=8.3 Hz, 2H), 7.39 (d, J=8.4 Hz, 2H), 3.85–4.06 (m, 6H), 3, 60–3.69 (m, 2H), 3.50–3.59 (m, 1H), 3.44 (br d, J=7.7 Hz, 1H), 3.14–3.31 (m , 2H), 2.21 (dt, J=8.4, 4.2 Hz, 1H), 1.94–2.12 (m, 4H), 1.76–1.93 (m, 1H), 1.07 (br t, J=7.1 Hz, 1H).

¹³ C NMR (DMSO–d6) δ: 165.0, 154.0, 143.5, 142.0, 140.1, 132.8, 127.7, 126.5, 124.2, 110.7, 65.8, 62.5, 55.6, 53.3, 53.3, 29.9, 28.9, 28.8, 23.6, 16.8.

MS(ESI–TOF): 394.2071 [M+H]+.

The starting material, 1R , 5S )-1-(4-bromophenyl)-3-(tetrahydro-2H-pyran-4-yl)-3-azabicyclo[3.1.0]hexan-3-ium chloride, was prepared as follows .

A 1 L volume of Redlay was loaded with (1 R .5 S )-1-(4-bromophenyl)-3-azabicyclo[3.1.0]hexan-3-ium chloride (30 g, 1 eq.), dihydro-2H-pyran –4( 3H )–one (13.13 g, 1.2 eq.) and THF (300 ml). The mixture was stirred at IT=22 ± 5°C for 1 hour. NaBH(OAc) ₃ (30.1 g, 1.3 eq.) was added in portions while maintaining IT < 30°C. The mixture was stirred at IT = 22 ± 5°C for 2 hours. 6.2% HCl (93 g, 90 ml, 1.5 eq.) was added to maintain IT < 30°C and pH < 2. The mixture was stirred for 10 min. 25% K ₂ CO ₃ (259 g, 210 ml) was added, adjusting pH=8-9. IPAc (300 ml) was added. The mixture was stirred for 10 minutes. After phase separation, H ₂ O (150 g) was added to the organic layer. The mixture was stirred for 10 minutes. After phase separation, the organic layer was concentrated under vacuum (50–100 mbar, water bath 50°C). IPA (120 g, 150 ml) was added. The organic layer was concentrated under vacuum (50–100 mbar, water bath 50°C). IPA (144 g, 180 ml) was added. The mixture was filtered through CMC. The filter cake was washed with IPA (24 g x 2). H ₂ O (5 g) was added to the organic layer. 31% HCl (19.3 g) was added dropwise at IT < 35°C. The mixture was stirred at IT=22 ± 5°C for 2 hours and filtered. The filter cake was washed with IPA (48 g x 2). The wet filter cake was dried under vacuum at 50°C for at least 6 hours. The desired product (31.4 g, 98% purity by HPLC, 78% yield) was obtained as a white solid.

¹ H NMR (DMSO–d6 and D ₂ O) δ: 7.46 (br d, J=8.4 Hz, 2H), 7.15 (br d, J=8.4 Hz, 2H), 3, 90 (br d, J=7.8 Hz, 3H), 3.62 (br s, 1H), 3.51 (br s, 2H), 3.16–3.38 (m, 3H), 2, 05–2.21 (m, 1H), 1.93 (br s, 2H), 1.49–1.71 (m, 2H), 1.05–1.30 (m, 1H).

¹³ C NMR (DMSO–d6) δ: 138.3, 131.9, 129.2, 120.4, 65.5, 62.3, 56.2, 53.9, 29.1, 28.9, 24.8, 23.0.

MS(ESI–TOF): 322.0761 [M+H]+.

The complete method for making Alternative Example 34A is depicted in the following Reaction Scheme 34A:

The first connection in this diagram, A1, can be made as follows.

Stage 1–2. Synthesis of 1c and 1d

Scheme 34A1

Reaction procedure (see also Xu, Feng et al., Org. Lett. Vol 8, No. 17, 2006, pages 3885–3888):

Add NaHMDS (2.0 M in THF, 19.2 L, 38.4 mol, 2.5 eq.) dropwise to solution 1a (3.00 kg, 15.3 mol, 1.0 eq. obtained, e.g. , from Sigma–Aldrich) and 1b (1.80 kg, 19.47 mmol, 1.27 eq; obtained, for example, from Sigma–Aldrich) in THF (30 L) at –15–20°C under N atmosphere ₂ for 5 hours. The reaction mixture was stirred for an additional 3 hours at –15°C, then gradually heated to room temperature. and stirred for 16 hours. HPLC showed that 4-bromophenylacetonitrile was completely consumed.

BF was added slowly₃.Et₂O (4.74 kg, 15.3 mol, 1.0 eq.) at 20–40°C. After addition, BH was added slowly₃⋅DMS (19.2 L, 38.4 mol, 2.5 eq.) at 20–40°C. After addition, the mixture was stirred at 50–55°C for 16 hours. HPLC showed that the intermediate was completely consumed.

The mixture was cooled to –5–0°C and AcOH (4.5 kg, 74.4 mol, 4.86 eq) was added slowly. After addition, 3N was added slowly. HCl (36 kg). The mixture was stirred for 1 hour and most of the THF was removed under vacuum at 40–45°C. The residue was then extracted with MTBE (3 x 10 L). The MTBE phase was washed with 3N. HCl (4x5 l). The combined aqueous layer was cooled to 0°C. The aqueous layer was then neutralized with 30% NaOH to pH >10, extracted with IPAc (3 x 15 kg), washed with brine (5 L), evaporated in vacuo to give crude 1d (3.71 kg) which was used directly to the next stage.

Stage 3–4. Synthesis of 1e and 1f

Scheme 34A2

Reaction procedure

Cool IPAc (12 L) to 0°C and add SOCl ₂ (5.46 kg, 45.9 mol, 1.5 eq) while maintaining temperature at 0–4°C. After addition, a solution of crude 1d (7.46 kg, two batches combined from 6 kg of 1a) in IPAc (24 L) was slowly added. After addition, the mixture was stirred for 1 hour at 0°C, then heated to room temperature. and stirred for 4 hours. HPLC showed that the intermediate was completely consumed.

The mixture was cooled to –5–0°C and quenched with water (30 L), then 30% aqueous NaOH was added slowly, adjusting pH to 8.5–9 and stirred overnight at 0–5°C. HPLC showed that the intermediate was completely consumed.

A 30% aqueous NaOH solution was slowly added, adjusting the pH to 11–12 at 0–10°C and stirred at room temperature. for 30 min. The phases were separated, the aqueous layer was extracted with IPAc (10 kg), the combined organic phase was washed with brine (5 L), dried over Na ₂ SO ₄ and filtered. The filtrate was then cooled to 0–5°C. 4 M HCl/IPA (8.0 kg, 151 mol) was added slowly at 0–5°C. After addition, the reaction mixture was stirred for 4 h at 0–5°C. The mixture was filtered, collected the solid, washed with MTBE (6 L) and dried under vacuum to give compound 1f (3.10 kg, analysis 98%, yield 37%)

Alternative Example 34B.

Starting material A5 in Scheme 34A can alternatively also be prepared as follows (Reaction Scheme 34B) and then used as shown in Alternate Example 34A above.

The detailed procedure is as follows.

In a 100 ml three-neck flask, a solution of (1 R .5 S )-1-(4-bromophenyl)-3-tetrahydropyran-4-yl-3-azabicyclo[3.1.0]hexane (5.00 g, 15.52 mmol, 1.00 eq.) in anhydrous THF (50.00 ml) to –78°C under N ₂ atmosphere. Add n –BuLi (2.5 M in hexane, 7.45 mL, 1.20 eq.) dropwise at –78°C and stir for 1 hour at –78°C. A solution of trimethyl borate (4.84 g, 46.56 mmol, 5.26 mL, 3.00 eq) in anhydrous THF (10.00 mL) was added dropwise at –78°C. After addition, the reaction mixture was heated to 25°C and stirred at 25°C for 16 hours. After the starting material was completely consumed, LC–MS was monitored and 74.23% of the target compound was detected. The reaction mixture was cooled to 0°C in an ice bath and quenched with saturated NH ₄ Cl solution (50 ml). Adjust the pH of the mixture to 12–13 using NaOH solution (1 M) and extract it with EtOAc (30 ml x 3). The aqueous layer was concentrated under reduced pressure to obtain a residue (18 g). The residue was purified by prep-HPLC (TFA conditions) to give [4-[( 1R , 5S )-3-tetrahydropyran-4-yl-3-azabicyclo[3.1.0]hexane-1-yl]phenyl] boronic acid (4.60 g, 11.47 mmol, 73.88% yield, TFA salt) as a white solid.

^1H NMR (400 MHz, DMSO–d6) δ: 7.72 (d, 2H), 7.24 (d, 2H), 4.13 (d, 1H), 4.06–4.03 (m, 2H), 3.86 (d, 1H), 3.70–3.67 (m, 2H), 3.45–3.32 (m, 3H), 2.26–2.23 (m, 1H) , 2.11–2.05 (m, 2H), 1.81–1.77 (m, 2H), 1.34–1.20 (m, 2H). ESI–MS 288 [M+H] ⁺ .

A mixture of [4-[(1 R .5 S )-3-tetrahydropyran-4-yl-3-azabicyclo[3.1.0]hexan-1-yl]phenyl]boronic acid (500.00 mg, 1.25 mmol) was heated , 1.00 equiv TFA salt), methyl 2-amino-5-bromopyridine-3-carboxylate (433.22 mg, 1.88 mmol, 1.50 equiv.), Pd(dppf)Cl ₂ (91, 46 mg, 0.125 mmol, 0.10 eq.) and K ₂ CO ₃ (518.29 mg, 3.75 mmol, 3.00 eq.) in dioxane (5.00 ml) and H ₂ O (1.00 ml) to 100°C and stirred for 16 hours at 100°C under N ₂ atmosphere. The reaction mixture was cooled to 30°C and concentrated under reduced pressure. The residue was purified by prep-HPLC (neutral conditions) to give methyl 2-amino-5-[4-[( 1R , 5S )-3-tetrahydropyran-4-yl-3-azabicyclo[3.1.0]hexane –1-yl]phenyl]pyridine-3-carboxylate (100.00 mg, 20.02% yield, 98.47% HPLC purity) as a brown solid.

^1H NMR (400 MHz, DMSO–d6) δ: 8.49 (d, 1H), 8.35 (d, 1H), 7.46 (d, 2H), 7.24 (d, 2H), 4 .01–3.98 (m, 2H), 3.94 (s, 3H), 3.47–3.41 (m, 3H), 3.19 (d, 1H), 2.65 (d, 1H ), 2.50–2.55 (m, 1H), 2.45–2.35 (m, 1H), 1.81–1.76 (m, 3H), 1.64–1.46 (m , 2H), 1.45–1.40 (m, 1H), 0.86–0.83 (m, 1H). ESI–MS 394 [M+H] ⁺ .

Example 35 : 2-amino-5-(4-((1S,5R)-3-(4,4-difluorocyclohexyl)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)-N-(4 –hydroxybicyclo[2.2.2]octan-1-yl)nicotinamide

The title compound was prepared in a manner similar to (1R,5S) -tert -butyl-1-(4-(6-amino-5-(((1r,4R)-4-hydroxycyclohexyl)carbamoyl)pyridin-3-yl )phenyl)-3-azabicyclo[3.1.0]hexane-3-carboxylate (intermediate 1b), except that 2-amino-5-(4-((1S,5R)-3-(4, 4-difluorocyclohexyl)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)nicotinic acid (intermediate 35a) instead of 2-amino-5-(4-((1R,5S)-3-( tert - butoxycarbonyl)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)nicotinic acid (intermediate 1c) and 4-aminobicyclo[2.2.2]octan-1-ol hydrochloride was used instead of trans -4-aminocyclohexanol. The crude product was purified by prepHPLC (Method 1a) to obtain the title compound as the TFA salt. ^1H NMR (400 MHz, DMSO–d6) δ 9.05 (bs, 1H), 8.42 (d, 1H), 8.22 (s, 1H), 8.02 (s, 1H), 7. 69 (d, 2H), 7.50 (bs, 2H), 7.39 (d, 2H), 4.83 (bs, 1H), 4.08 (dd, 1H), 3.80–3.25 (m, 6H), 2.31–2.09 (m, 4H), 2.09–1.99 (m, 6H), 1.98–1.71 (m, 4H), 1.70–1 .56 (m, 6H), 1.42–1.40 (m, 1H), 1.10 (t, 1H). (UPLC–MS) t _R 0.62 min; ESI–MS 537 [M+H] ⁺ .

Intermediate 35a : 2-amino-5-(4-((1S,5R)-3-(4,4-difluorocyclohexyl)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)nicotinic acid

The title compound was prepared in a manner similar to that for 2-amino-5-(4-((1R,5S)-3-( tert -butoxycarbonyl)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)nicotinic acid (intermediate 2c), except that methyl-2-amino-5-(4-((1S,5R)-3-(4,4-difluorocyclohexyl)-3-azabicyclo[3.1.0]hexane) was used –1-yl)phenyl)nicotinate (intermediate 35b) instead of (1R,5S)- tert -butyl-1-(4-(6-amino-5-(methoxycarbonyl)pyridin-3-yl)phenyl)-3- azabicyclo[3.1.0]hexane-3-carboxylate (intermediate 1d). (UPLC–MS) t _R 0.46 min; ESI–MS 414 [M+H] ⁺ .

Intermediate 35b : Methyl-2-amino-5-(4-((1S,5R)-3-(4,4-difluorocyclohexyl)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)nicotinate

The title compound was prepared in a manner similar to that for methyl-2-amino-5-(4-((1R,5S)-3-(tetrahydro-2H-pyran-4-yl)-3-azabicyclo[3.1.0]hexane -1-yl)phenyl)nicotinate (intermediate 14b), except that methyl 5-(4-((1S,5R)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl) was used –2-aminonicotinate hydrochloride (intermediate 7c) instead of methyl 5-(4-((1R,5S)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)-2-aminonicotinate hydrochloride (intermediate 14c ) and used 4,4-difluorocyclohexanone instead of dihydro-2H-pyran-4(3H)-one. (UPLC–MS) t _R 0.73 min; ESI–MS 428 [M+H] ⁺ .

Example 36 : 2-amino-N-(3-(hydroxymethyl)bicyclo[1.1.1]pentan-1-yl)-5-(4-((1R,5S)-3-isopropyl-3-azabicyclo[3.1. 0]hexan–1–yl)phenyl)nicotinamide

The title compound was prepared in a manner similar to that for 2-amino-N-((1r,4R)-4-hydroxycyclohexyl)-5-(4-((1R,5S)-3-isopropyl-3-azabicyclo[3.1.0 ]hexan-1-yl)phenyl)nicotinamide (Example 2), except that (3-aminobicyclo[1.1.1]pentan-1-yl)methanol (intermediate 16a) was used instead of trans -4-aminocyclohexanol hydrochloride. ^1H NMR (400 MHz, DMSO–d6) δ 9.05 (s, 1H), 8.41 (d,1H), 8.18 (d,1H), 7.61 (d, 2H), 7, 48 (dd, 1H), 7.33–7.17 (m, 4H), 4.55 (t, 1H), 3.51 (d, 3H), 3.31 (s, 2H), 3.08 (s, 2H), 2.07–1.99 (m, 1H), 1.41–1.22 (m, 3H), 1.17–0.98 (m, 9H), 0.79 (d , 1H). (UPLC–MS) t _R 0.54 min; ESI–MS 433 [M+H] ⁺ .

Example 37 : 2-amino-5-(4-((1S,5R)-3-(but-2-in-1-yl)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)- N–((1r,4S)–4–hydroxycyclohexyl)nicotinamide

The title compound was prepared in a manner similar to that for 2-amino-5-(4-((1R,5S)-3-(1,3-difluoropropan-2-yl)-3-azabicyclo[3.1.0]hexane-1 -yl)phenyl)-N-((1r,4R)-4-hydroxycyclohexyl)nicotinamide (Example 5), except that 1-bromobut-2-yne was used instead of 4-(3-bromopropyl)morpholine and the reaction mixture stirred at room temperature. within 30 min. ^1H NMR (400 MHz, DMSO–d6) δ 8.38 (s, 1H), 8.31 (d, 1H), 8.12 (d, 1H), 7.59 (d, 2H), 7. 21 (s, 2H), 7.13 (s, 2H), 4.60 (d, 1H), 3.72 (s, 1H), 3.39 (s, 3H), 3.21 (s, 1H ), 2.98–2.58 (m, 3H), 1.84 (dd, 8H), 1.49–1.17 (m, 5H), 0.78 (s, 1H). (UPLC–MS) t _R 0.54 min; ESI–MS 445 [M+H] ⁺ .

Example 38 : 2-amino-N-((1r,4S)-4-hydroxycyclohexyl)-5-(4-((1S,5R)-3-(oxetan-3-ylmethyl)-3-azabicyclo[3.1.0 ]hexan–1–yl)phenyl)nicotinamide

The title compound was prepared in a manner similar to that for 2-amino-5-(4-((1R,5S)-3-(1,3-difluoropropan-2-yl)-3-azabicyclo[3.1.0]hexane-1 -yl)phenyl)-N-((1r,4R)-4-hydroxycyclohexyl)nicotinamide (Example 5), except that 3-(iodomethyl)oxetane was used instead of 4-(3-bromopropyl)morpholine and the reaction mixture was stirred at 60°C for 60 min. ^1H NMR (400 MHz, DMSO–d6) δ 8.39 (d, 1H), 8.30 (d, 1H), 8.11 (d, 1H), 7.59 (d, 2H), 7. 22 (d, 2H), 7.12 (s, 2H), 4.73–4.61 (m, 2H), 4.58 (d, 1H), 4.29 (s, 2H), 3.84 –3.67 (m, 1H), 3.52–3.38 (m, 2H), 3.27–3.12 (m, 2H), 2.94 (d, 1H), 2.80 (s , 2H), 1.95–1.76 (m, 5H), 1.54–1.14 (m, 6H), 0.76 (s, 1H). (UPLC–MS) t _R 0.49 min; ESI–MS 463 [M+H] ⁺ .

Example 39 : 2-amino-N-((1r,4R)-4-hydroxycyclohexyl)-5-(4-((1R,5S)-3-(1-(2,2,2-trifluoroethyl)piperidine-4 –yl)-3-azabicyclo[3.1.0]hexane-1-yl)phenyl)nicotinamide

The title compound was prepared in a manner similar to that for 2-amino-N-((1r,4R)-4-hydroxycyclohexyl)-5-(4-((1R,5S)-3-(1-(methylsulfonyl)piperidine-4 -yl)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)nicotinamide (Example 1), except that 1-(2,2,2-trifluoroethyl)piperidin-4-one was used instead of 1 –(methylsulfonyl)piperidin-4-one. ^1H NMR (400 MHz, DMSO–d6) δ 8.38 (d, 1H), 8.31 (d,1H), 8.11 (d, 1H), 7.58 (d, 2H), 7. 23 (d, 2H), 7.12 (s, 2H), 4.55 (bs, 1H), 3.80–3.62 (m, 1H), 3.20–3.04 (m, 3H) , 2.88 (d, 2H), 2.57 (d, 1H), 2.46 (dd, 1H), 2.41–2.29 (m, 2H), 2.15 (t, 1H), 1.99–1.69 (m, 8H), 1.49–1.17 (m, 7H), 1.15 (t, 1H), 0.75 (dd, 1H). (UPLC–MS) t _R 0.62 min; ESI–MS 558 [M+H] ⁺ .

Example 40 : 2-amino-N-(4-hydroxybicyclo[2.2.2]octan-1-yl)-5-(4-((1S,5R)-3-(2-morpholinoethyl)-3-azabicyclo[3.1 .0]hexan–1–yl)phenyl)nicotinamide

The title compound was prepared in a manner similar to that for 2-amino-5-(4-((1R,5S)-3-(1,3-difluoropropan-2-yl)-3-azabicyclo[3.1.0]hexane-1 -yl)phenyl)-N-((1r,4R)-4-hydroxycyclohexyl)nicotinamide (Example 5), except that the HCl salt and 5-(4-((1S,5R)-3-azabicyclo[ 3.1.0]hexan-1-yl)phenyl)-2-amino-N-(4-hydroxybicyclo[2.2.2]octan-1-yl)nicotinamide (intermediate 40a) instead of 5-(4-((1S, 5R)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)-2-amino-N-((1r,4R)-4-hydroxycyclohexyl)nicotinamide (intermediate 5a) and 4-(2- bromoethyl)morpholine instead of 4-(3-bromopropyl)morpholine, and the reaction mixture was stirred at 60°C for 3 hours. ^1H NMR (400 MHz, DMSO–d6) δ 8.34 (d, 1H), 7.98 (d, 1H), 7.79 (s, 1H), 7.57 (d, 2H), 7.21 (d, 2H), 6.92 (s, 2H), 4.32 (s, 1H) , 3.65–3.49 (m, 4H), 3.08 (d, 1H), 2.61 (t, 2H), 2.59–2.39 (m, 9H), 2.15–1 .98 (m, 6H), 1.85–1.73 (m, 1H), 1.70–1.54 (m, 6H), 1.31 (t, 1H), 0.75 (dd, 1H ). (UPLC–MS) t _R 0.52 min; ESI–MS 532 [M+H] ⁺ .

Intermediate 40a : 5–(4–((1S,5R)–3–azabicyclo[3.1.0]hexan–1–yl)phenyl)–2–amino–N–(4–hydroxybicyclo[2.2.2]octane– 1-yl)nicotinamide

The title compound was prepared in a manner similar to that for 5-(4-((1S,5R)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)-2-amino-N-((1r,4R) –4-hydroxycyclohexyl)nicotinamide (intermediate 5a), except that (1S,5R) -tert -butyl-1-(4-(6-amino-5-((4-hydroxybicyclo[2.2.2] octan-1-yl)carbamoyl)pyridin-3-yl)phenyl)-3-azabicyclo[3.1.0]hexane-3-carboxylate (intermediate 40b) instead of (1S,5R) -tert -butyl-1-(4 –(6-amino-5-(((1r,4S)-4-hydroxycyclohexyl)carbamoyl)pyridin-3-yl)phenyl)-3-azabicyclo[3.1.0]hexane-3-carboxylate (intermediate 5b). One fraction of the title compound was obtained as the hydrochloride salt after evaporation of the dioxane; the other fraction was obtained as the TFA salt after further purification by prepHPLC (method 1a). (UPLC–MS) t _R 0.48 min; ESI–MS 419 [M+H] ⁺ .

Intermediate 40b : (1S,5R)- tert -butyl-1-(4-(6-amino-5-((4-hydroxybicyclo[2.2.2]octan-1-yl)carbamoyl)pyridin-3-yl) phenyl)-3-azabicyclo[3.1.0]hexane-3-carboxylate

The title compound was prepared in a manner similar to (1R,5S) -tert -butyl-1-(4-(6-amino-5-(((1r,4R)-4-hydroxycyclohexyl)carbamoyl)pyridin-3-yl )phenyl)-3-azabicyclo[3.1.0]hexane-3-carboxylate (intermediate 1b), except that 2-amino-5-(4-((1S,5R)-3-( tert - butoxycarbonyl)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)nicotinic acid (intermediate 5c) instead of 2-amino-5-(4-((1R,5S)-3-( tert -butoxycarbonyl) –3-azabicyclo[3.1.0]hexan-1-yl)phenyl)nicotinic acid (intermediate 1c) and used 4-aminobicyclo[2.2.2]octan-1-ol hydrochloride instead of trans –4-aminocyclohexanol. (UPLC–MS) t _R 1.05 min; ESI–MS 519 [M+H] ⁺ .

Example 41 : 2-amino-N-((1r,4R)-4-hydroxycyclohexyl)-5-(4-((1R,5S)-3-(2-(2,2,2-trifluoroethoxy)ethyl)- 3-azabicyclo[3.1.0]hexane-1-yl)phenyl)nicotinamide

The title compound was prepared in a manner similar to that for 2-amino-5-(4-((1R,5S)-3-(1,3-difluoropropan-2-yl)-3-azabicyclo[3.1.0]hexane-1 -yl)phenyl)-N-((1r,4R)-4-hydroxycyclohexyl)nicotinamide (Example 6), except that 2-(2,2,2-trifluoroethoxy)ethyl trifluoromethanesulfonate (intermediate 41a) was used instead of 1 ,3-difluoropropane-2-yltrifluoromethanesulfonate (intermediate 6a). After purification, the title compound was obtained as the TFA salt. ^1H NMR (400 MHz, DMSO–d6) δ 9.15 (bs, 1H), 8.40 (s, 1H), 8.31 (d, 1H), 8.12 (d, 1H), 7, 63 (d, 2H), 7.23 (s, 2H), 7.13 (s, 2H), 4.59 (d, 1H), 4.11 (s, 2H), 3.80–3.63 (m, 3H), 3.40 (m, 4H), 2.75–2.60 (m, 3H), 1.86 (m, 5H), 1.51–1.15 (m, 6H). (UPLC–MS) t _R 0.60 min; ESI–MS 519 [M+H] ⁺ .

Intermediate 41a : 2-(2,2,2-trifluoroethoxy)ethyl trifluoromethanesulfonate

The title compound was prepared in a manner similar to 1,3-difluoropropane-2-yltrifluoromethanesulfonate (intermediate 6a), except that 2-(2,2,2-trifluoroethoxy)ethanol was used instead of 1,3-difluoropropane-2 -ol, and it was obtained as a crude oil, which was used without further purification.

Example 42 : 2-amino-N-(4-hydroxybicyclo[2.2.2]octan-1-yl)-5-(4-((1S,5R)-3-(tetrahydro-2H-pyran-4-yl) –3-azabicyclo[3.1.0]hexane-1-yl)phenyl)nicotinamide

The title compound was prepared in a manner similar to that for 2-amino-N-((1r,4R)-4-hydroxycyclohexyl)-5-(4-((1R,5S)-3-(1-(methylsulfonyl)piperidine-4 -yl)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)nicotinamide (Example 1), except that the HCl salt and 5-(4-((1S,5R)-3-azabicyclo [3.1.0]hexan-1-yl)phenyl)-2-amino-N-(4-hydroxybicyclo[2.2.2]octan-1-yl)nicotinamide (intermediate 40a) instead of 5-(4-((1R ,5S)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)-2-amino-N-((1r,4R)-4-hydroxycyclohexyl)nicotinamide (intermediate 1a) and dihydro-2H- pyran-4(3H)-one instead of 1-(methylsulfonyl)piperidin-4-one. After purification, the title compound was obtained as the TFA salt. 1H NMR (400 MHz, DMSO–d6) δ 9.62 (bs, 1H), 8.38 (d, 1H), 7.98 (d, 1H), 7.81 (s, 1H), 7.65 (d, 2H), 7.37 (d, 2H), 6.95 (s, 2H), 4.33 (s, 1H), 4.06–3.97 (m, 3H), 3.75– 3.43 (m, 4H), 3.29–3.26 (m, 2H), 2.28–2.19 (m, 1H), 2.07–2.03 (m, 6H), 2. 03–1.96 (m, 2H), 1.75–1.63 (m, 1H), 1.63–1.61 (m, 6H), 1.63–1.61 (m, 1H), 1.25–1.17 (m, 1H). (UPLC–MS) t _R 0.52 min; ESI–MS 503 [M+H] ⁺ .

Example 43 : 2-amino-N-cyclohexyl-5-(4-(3-(piperidin-1-yl)propoxy)phenyl)nicotinamide

1-(3-(4-(4 ,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenoxy)propyl)piperidine (60.9 mg, 0.168 mmol), K ₂ CO ₃ (69.5 mg, 0.503 mmol) and PdCl ₂ (PPh ₃ ) ₂ (11.8 mg, 0.017 mmol). The reaction mixture was stirred for 15 minutes. at 80°C, then cooled and diluted with 5 ml EtOAc and filtered through a pad of Na ₂ SO ₄ . After concentration under reduced pressure, the crude product was purified by prepHPLC (Method 1a) to obtain the title compound as a colorless solid. ^1H NMR (400 MHz, DMSO–d6) δ 8.34 (d, 1H), 8.33 (d, 1H), 8.09 (d, 1H), 7.59 (d, 2H), 7. 06 (s, 2H), 7.01 (d, 2H), 4.03 (t, 2H), 3.75 (s, 1H), 2.50–2.22 (m, 6H), 1.95 –1.65 (m, 6H), 1.62 (d, 1H), 1.52–1.48 (m, 4H), 1.46–1.34 (m, 6H), 1.32–1 .15 (m, 1H). (HPLC–MS) t _R 1.46 min; APCI–MS 437.3 [M+H] ⁺ .

Intermediate 43a : 2-amino-5-bromo-N-cyclohexylnicotinamide

The title compound was prepared in a manner similar to (1R,5S) -tert -butyl-1-(4-(6-amino-5-(((1r,4R)-4-hydroxycyclohexyl)carbamoyl)pyridin-3-yl )phenyl)-3-azabicyclo[3.1.0]hexane-3-carboxylate (intermediate 1b), except that 2-amino-5-bromonicotinic acid was used instead of 2-amino-5-(4-((1R ,5S)-3-( tert -butoxycarbonyl)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)nicotinic acid (intermediate 1c) and used cyclohexanamine instead of trans -4-aminocyclohexanol. The crude product was used without further purification. (HPLC–MS) t _R 0.98 min; ESI–MS 208/300 [M+H] ⁺ .

Example 44 : 2-amino-5-(4-((1R,5S)-3-(1,1-dioxidotetrahydro-2H-thiopyran-4-yl)-3-azabicyclo[3.1.0]hexan-1-yl )phenyl)–N–((1r,4R)–4–hydroxycyclohexyl)nicotinamide

The title compound was prepared in a manner similar to that for 2-amino-N-((1r,4R)-4-hydroxycyclohexyl)-5-(4-((1R,5S)-3-(1-(methylsulfonyl)piperidine-4 –yl)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)nicotinamide (1), except that dihydro-2H-thiopyran-4(3H)-one-1,1-dioxide was used instead 1-(methylsulfonyl)piperidin-4-one. ^1H NMR (400 MHz, DMSO–d6) δ 8.39 (d, 1H), 8.31 (d, 1H), 8.12 (d, 1H), 7.59 (d, 2H), 7. 24 (d, 2H), 7.12 (s, 2H), 4.58 (d, 1H), 3.82–3.65 (m, 1H), 3.48–3.35 (m, 2H) , 3.19–2.91 (m, 5H), 2.62–2.51 (m, 2H), 2.17–1.97 (m, 4H), 1.89–1.83 (m, 5H), 1.45–1.19 (m, 6H), 0.81 (dd, 1H). (UPLC–MS) t _R 0.48 min; ESI–MS 525 [M+H] ⁺ .

Example 45 : 2-amino-N-cyclohexyl-5-(4-(4-methylpiperazine-1-carbonyl)phenyl)nicotinamide

The title compound was prepared in a manner similar to that for 2-amino-N-cyclohexyl-5-(4-(3-(piperidin-1-yl)propoxy)phenyl)nicotinamide (Example 43), except that (4 –methylpiperazin-1-yl)-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)methanone instead of 1-(3-(4-(4,4 ,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenoxy)propyl)piperidine. ^1H NMR (400 MHz, DMSO–d6) δ 8.46 (d, 1H), 8.37 (s, 1H), 8.21 (d, 1H), 7.76 (d, 2H), 7. 47 (d, 2H), 7.22 (s, 2H), 3.82–3.55 (m, 1H), 3.35 (t, 4H), 2.31–2.22 (t, 4H) , 2.21 (s, 3H), 1.90–1.80 (m, 2H), 1.80–1.70 (m, 2H), 1.70–1.55 (m, 1H), 1 .35–1.29 (m, 4H), 1.28–1.05 (m, 1H). (HPLC–MS) t _R 1.29 min; APCI–MS 422 [M+H] ⁺ .

Example 46 : 2-amino-5-(3-fluoro-4-(((R)-2-methylpyrrolidin-1-yl)methyl)phenyl)-N-((1r,4R)-4-hydroxy-4- methylcyclohexyl)nicotinamide

The microwave vessel was loaded with 2-amino-5-bromo-N-((1r,4r)-4-hydroxy-4-methylcyclohexyl)nicotinamide (intermediate 59b, 33 mg, 0.10 mmol), (R)- 2-(3-fluoro-4-((2-methylpyrrolidin-1-yl)methyl)phenyl)-6-methyl-1,3,6,2-dioxazaborocane-4,8-dione (intermediate 46a, 38, 3 mg, 0.11 mmol) and XPhos Pd (7.39 mg, 0.01 mmol). Dioxane (1 ml) was added followed by 3N. aq. K ₃ PO ₄ solution (0.10 ml, 0.30 mmol). The reaction mixture was purged with a stream of nitrogen for 5 min and then irradiated in a microwave reactor at 120°C for 30 min. After cooling, the reaction mixture was concentrated under reduced pressure and purified by prepHPLC (Method 1b) to give the title compound. UPLC–HRMS t _R 3.21 min; ESI 441.26 [M+H] ⁺ .

Intermediate 46a : (R)-2-(3-fluoro-4-((2-methylpyrrolidin-1-yl)methyl)phenyl)-6-methyl-1,3,6,2-dioxazaborocane-4,8- dione

Shake a solution of 2-fluoro-4-(6-methyl-4,8-dioxo-1,3,6,2-dioxazaborocan-2-yl)benzaldehyde (intermediate 46b, 112 mg, 0.40 mmol), (R )-(2)-methylpyrrolidine (37.5 mg, 0.44 mmol) and acetic acid (2.4 mg, 0.040 mmol) in THF (2 ml) for 2 hours at room temperature. and sodium triacetoxyborohydride (110 mg, 0.52 mmol) was added. The resulting reaction mixture was shaken for 18 hours and then filtered through an SPE carbonate cartridge, followed by washing with THF. The filtrate and washings were concentrated under reduced pressure to give the title compound, which was used without further purification. ESI–MS 349 [M+H] ⁺ .

Intermediate 46b : 2-fluoro-4-(6-methyl-4,8-dioxo-1,3,6,2-dioxazaborocan-2-yl)benzaldehyde

Stir a solution of (3-fluoro-4-formylphenyl)boronic acid (67 mg, 0.40 mmol) and N-methyliminodiacetic acid (65 mg, 0.44 mmol) in DMF (2 ml) at 90°C for 18 hours The reaction mixture was cooled and filtered through an SPE carbonate cartridge, followed by washing with acetonitrile three times. The filtrate and washings were concentrated under reduced pressure to give the title compound, which was used without further purification. ESI–MS 558 [2M+H] ⁺ .

Example 47 : 2-amino-N-((1r,4R)-4-hydroxycyclohexyl)-5-(4-((1R,5S)-3-(3,3,3-trifluoropropyl)-3-azabicyclo[3.1 .0]hexan–1–yl)phenyl)nicotinamide

The title compound was prepared in a manner similar to that for 2-amino-5-(4-((1R,5S)-3-(1,3-difluoropropan-2-yl)-3-azabicyclo[3.1.0]hexane-1 -yl)phenyl)-N-((1r,4R)-4-hydroxycyclohexyl)nicotinamide (Example 6), except that 3,3,3-trifluoropropyltrifluoromethanesulfonate was used instead of 1,3-difluoropropan-2-yltrifluoromethanesulfonate (intermediate compounds 6a). ^1H NMR (400 MHz, DMSO–d6) δ 8.39 (s, 1H), 8.31 (d, 1H), 8.12 (d, 1H), 7.60 (d, 2H), 7. 23 (d, 2H), 7.13 (s, 2H), 4.59 (d, 1H), 3.74 (m, 1H), 3.40 (m, 3H), 3.09 (m, 1H ), 2.72–2.64 (m, 4H), 1.86 (m, 5H), 1.49–1.14 (m, 6H), 0.78 (s, 1H). (UPLC–MS) t _R 0.60 min; ESI–MS 489 [M+H] ⁺ .

Example 48 : 2-amino-5-(4-((1R,5S)-3-(2-fluoroethyl)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)-N-((1r, 4R)–4-hydroxycyclohexyl)nicotinamide

The title compound was prepared in a manner similar to that for 2-amino-5-(4-((1R,5S)-3-(1,3-difluoropropan-2-yl)-3-azabicyclo[3.1.0]hexane-1 -yl)phenyl)-N-((1r,4R)-4-hydroxycyclohexyl)nicotinamide (Example 6), except that 1-bromo-2-fluoroethane was used instead of 1,3-difluoropropan-2-yltrifluoromethanesulfonate (intermediate compound 6a) and the reaction mixture was stirred at 60°C for 3 hours. After purification, the title compound was obtained as the TFA salt. ^1H NMR (400 MHz, DMSO–d6) δ 9.95 (bs, 1H), 8.41 (s, 1H), 8.32 (d, 1H), 8.13 (d, 1H), 7. 70–7.57 (m, 2H), 7.36 (bs, 2H), 7.16 (s, 2H), 4.80–4.62 (m, 2H), 4.59 (d, 1H) , 4.08 (bs, 1H), 3.77–3.52 (m, 4H), 3.51–3.35 (m, 3H), 2.24 (bs, 1H), 1.87 (t , 4H), 1.59–1.13 (m, 6H). (UPLC–MS) t _R 0.46 min; ESI–MS 439 [M+H] ⁺ .

Example 49 : 2-amino-N-cyclohexyl-5-(4-morpholinophenyl)nicotinamide

The title compound was prepared in a manner similar to 2-amino-N-cyclohexyl-5-(4-(3-(piperidin-1-yl)propoxy)phenyl)nicotinamide (Example 43), except that 4- (4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)morpholine instead of 1-(3-(4-(4,4,5,5-tetramethyl-1 ,3,2-dioxaborolan-2-yl)phenoxy)propyl)piperidine. ^1H NMR (400 MHz, DMSO–d6) δ 8.35 (d, 1H), 8.34 (s, 1H), 8.08 (d, 1H), 7.55 (d, 2H), 7. 03 (d, 2H), 7.03 (s, 2H), 3.85–3.65 (m, 1H), 3.77 (t, 4H), 3.14 (t, 4H), 1.90 –1.80 (m, 2H), 1.80–1.70 (m, 2H), 1.80 (dd, 1H), 1.38–1.20 (m, 4H), 1.19–1 .05 (m, 1H). (HPLC–MS) t _R 1.60 min; APCI–MS 381 [M+H] ⁺ .

Example 50 : 2-amino-5-(4-((1R,5S)-3-(2,2-difluoropropyl)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)-N-(( 1r,4R)–4–hydroxycyclohexyl)nicotinamide

The title compound was prepared in a manner similar to that for 2-amino-5-(4-((1R,5S)-3-(1,3-difluoropropan-2-yl)-3-azabicyclo[3.1.0]hexane-1 -yl)phenyl)-N-((1r,4R)-4-hydroxycyclohexyl)nicotinamide (Example 6), except that 2,2-difluoropropyltrifluoromethanesulfonate was used instead of 1,3-difluoropropane-2-yltrifluoromethanesulfonate (intermediate 6a ) and the reaction mixture was stirred at room temperature. within 60 min. ^1H NMR (400 MHz, DMSO–d6) δ 8.39 (s, 1H), 8.33 (s, 1H), 8.14 (s, 1H), 7.59 (d, 2H), 7. 32–7.11 (m, 4H), 4.58 (bs, 1H), 3.74 (bs, 1H), 3.11 (bs, 1H), 3.00–2.66 (m, 5H) , 1.86 (bs, 5H), 1.63 (t, 3H), 1.48–1.18 (m, 6H), 0.81 (bs, 1H). (UPLC–MS) t _R 0.81 min; ESI–MS 471 [M+H] ⁺ .

Example 51 : 2-amino-5-(4-((1S,5R)-3-(2-cyanoethyl)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)-N-((1r, 4S)–4-hydroxycyclohexyl)nicotinamide

The title compound was prepared in a manner similar to that for 2-amino-5-(4-((1R,5S)-3-(1,3-difluoropropan-2-yl)-3-azabicyclo[3.1.0]hexane-1 -yl)phenyl)-N-((1r,4R)-4-hydroxycyclohexyl)nicotinamide (Example 5), except that 3-bromopropanenitrile was used instead of 4-(3-bromopropyl)morpholine and the reaction mixture was stirred at 60° C for 24 h. ¹ H NMR (400 MHz, DMSO–d6) δ 8.39 (d, 1H), 8.30 (d, 1H), 8.12 (d, 1H), 7.60 (d , 2H), 7.23 (d, 2H), 7.12 (s, 2H), 4.59 (d, 1H), 3.73 (d, 1H), 3.18 (d, 1H), 3 .09 (d, 1H), 2.81–2.62 (m, 6H), 1.99–1.78 (m, 5H), 1.48–1.18 (m, 6H), 0.80 (dd, 1H). (UPLC–MS) t _R 0.50 min; ESI–MS 446 [M+H] ⁺ .

Example 52 : 2-amino-N-cyclohexyl-5-(4-(3-morpholinopropoxy)phenyl)nicotinamide

The title compound was prepared in a manner similar to 2-amino-N-cyclohexyl-5-(4-(3-(piperidin-1-yl)propoxy)phenyl)nicotinamide (Example 43), except that 4- (3–(4–(4,4,5,5–tetramethyl–1,3,2–dioxaborolan–2–yl)phenoxy)propyl)morpholine instead of 1–(3–(4–(4,4,5, 5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenoxy)propyl)piperidine. ^1H NMR (400 MHz, DMSO–d6) δ 8.35 (d, 1H), 8.34 (s, 1H), 8.10 (d, 1H), 7.59 (d, 2H), 7. 06 (s, 2H), 7.01 (d, 2H), 4.04 (t, 2H), 3.82–3.68 (m, 1H), 3.58 (t, 4H), 2.50 –2.22 (m, 6H), 1.90–1.78 (m, 4H), 1.78–1.69 (m, 2H), 1.63 (dd, 1H), 1.38–1 .20 (m, 4H), 1.19–1.05 (m, 1H). (HPLC–MS) t _R 1.40 min; APCI–MS 439 [M+H] ⁺ .

Example 53 : 2-amino-N-((1r,4S)-4-hydroxycyclohexyl)-5-(4-((1S,5R)-3-(prop-2-in-1-yl)-3-azabicyclo [3.1.0]hexan–1–yl)phenyl)nicotinamide

The title compound was prepared in a manner similar to that for 2-amino-N-((1r,4S)-4-hydroxycyclohexyl)-5-(4-((1S,5R)-3-(3-morpholinopropyl)-3-azabicyclo [3.1.0]hexan-1-yl)phenyl)nicotinamide (Example 5), except that 3-bromoprop-1-yne was used instead of 4-(3-bromopropyl)morpholine. ^1H NMR (400 MHz, DMSO–d6) δ 8.38 (d, 1H), 8.31 (d, 1H), 8.12 (d, 1H), 7.59 (d, 2H), 7. 21 (d, 2H), 7.13 (s, 2H), 4.59 (d, 1H), 3.73 (d, 1H), 3.52–3.38 (m, 2H), 3.21 (d, 1H), 2.93 (d, 1H), 2.83 (d, 1H), 2.72 (d, 1H), 2.40 (s, 3H), 1.95–1.75 ( m, 4H), 1.44–1.19 (m, 5H), 0.80 (dd, 1H). (UPLC–MS) t _R 0.49 min; ESI–MS 431 [M+H] ⁺ .

Example 54 : 2-amino-N-((1r,4R)-4-hydroxycyclohexyl)-5-(4-((1R,5S)-3-(prop-2-in-1-yl)-3-azabicyclo [3.1.0]hexan–1–yl)phenyl)nicotinamide

The title compound was prepared in a manner similar to that for 2-amino-5-(4-((1R,5S)-3-(1,3-difluoropropan-2-yl)-3-azabicyclo[3.1.0]hexane-1 -yl)phenyl)-N-((1r,4R)-4-hydroxycyclohexyl)nicotinamide (Example 6), except that 3-bromoprop-1-yne was used instead of 1,3-difluoropropan-2-yltrifluoromethanesulfonate (intermediate compounds 6a). ^1H NMR (400 MHz, DMSO–d6) δ 8.39 (d, 1H), 8.31 (d, 1H), 8.12 (d, 1H), 7.59 (d, 2H), 7. 21 (d, 2H), 7.12 (s, 2H), 4.59 (d, 1H), 3.76–3.70 (m, 1H), 3.46 (s, 2H), 3.45 –3.37 (m, 1H), 3.22 (d, 1H), 2.83 (d, 1H), 2.72 (d, 1H), 2.71–2.69 (m, 2H), 1.92–1.77 (m, 5H), 1.51–1.17 (m, 5H), 0.80 (dd, 1H). (UPLC–MS) t _R 0.52 min; ESI–MS 431 [M+H] ⁺ .

Example 55 : 5-(4-((R)-1-(2-oxa-6-azaspiro[3.3]heptan-6-yl)ethyl)-2-chlorophenyl)-2-amino-N-((1r, 4R)–4-hydroxycyclohexyl)nicotinamide

The title compound was prepared in a manner similar to that for 5-(4-((R)-1-(2-oxa-6-azaspiro[3.3]heptan-6-yl)ethyl)phenyl)-2-amino-N-( (1r,4R)-4-hydroxycyclohexyl)nicotinamide (Example 3), except that (R)-6-(1-(4-bromo-3-chlorophenyl)ethyl)-2-oxa-6-azaspiro was used [3.3]heptane (intermediate 55a) instead of (R)-6-(1-(4-bromophenyl)ethyl)-2-oxa-6-azaspiro[3.3]heptane (intermediate 3b). ^1H NMR (400 MHz, DMSO–d6) δ 8.17 (m, 2H), 8.00 (s, 1H), 7.42–7.44 (m, 2H), 7.40 (m, 1H ), 7.34 (bs, 2H), 4.62 (s, 3H), 4.55–4.57 (m, 1H), 3.72 (m, 1H), 3.35–3.54 ( m, 4H), 3.20–3.24 (m, 2H), 3.15 (bs, 1H), 1.78–1.88 (m, 4H), 1.20–1.41 (m, 4H), 1.11 (bs, 3H). (UPLC–MS) t _R 0.47 min; ESI–MS 471/473 [M+H] ⁺ .

Intermediate 55a : (R)-6-(1-(4-bromo-3-chlorophenyl)ethyl)-2-oxa-6-azaspiro[3.3]heptane

The title compound was prepared in a manner similar to (R)-6-(1-(4-bromophenyl)ethyl)-2-oxa-6-azaspiro[3.3]heptane (intermediate 3b), except that ( R)-1-(4-bromo-3-chlorophenyl)ethanamine instead of (R)-1-(4-bromophenyl)ethanamine. (UPLC–MS) t _R 0.63 min; ESI–MS 316/318 [M+H] ⁺ .

Example 56 : 2-amino-5-(2-fluoro-4-(((R)-2-methylpyrrolidin-1-yl)methyl)phenyl)-N-((1r,4R)-4-hydroxy-4- methylcyclohexyl)nicotinamide

The title compound was prepared in a manner similar to that for 2-amino-5-(3-fluoro-4-(((R)-2-methylpyrrolidin-1-yl)methyl)phenyl)-N-((1r,4R)- 4-hydroxy-4-methylcyclohexyl)nicotinamide (Example 46), except that (R)-2-(2-fluoro-4-((2-methylpyrrolidin-1-yl)methyl)phenyl)-6- methyl-1,3,6,2-dioxazaborocane-4,8-dione (intermediate 56a) instead of (R)-2-(3-fluoro-4-((2-methylpyrrolidin-1-yl)methyl)phenyl) –6-methyl-1,3,6,2-dioxazaborocane-4,8-dione (intermediate 46a). UPLC–HRMS t _R 3.22 min; ESI 441.26 [M+H] ⁺ .

Intermediate 56a : (R)-2-(2-fluoro-4-((2-methylpyrrolidin-1-yl)methyl)phenyl)-6-methyl-1,3,6,2-dioxazaborocane-4,8- dione

The title compound was prepared in a manner similar to that for (R)-2-(3-fluoro-4-((2-methylpyrrolidin-1-yl)methyl)phenyl)-6-methyl-1,3,6,2-dioxazaborocane –4,8-dione (intermediate 46a), except that 3-fluoro-4-(6-methyl-4,8-dioxo-1,3,6,2-dioxazaborocan-2-yl)benzaldehyde was used (intermediate 56b) instead of 2-fluoro-4-(6-methyl-4,8-dioxo-1,3,6,2-dioxazaborocan-2-yl)benzaldehyde (intermediate 46b). ESI–MS 349 [M+H] ⁺ .

Intermediate 56b : 3-fluoro-4-(6-methyl-4,8-dioxo-1,3,6,2-dioxazaborocan-2-yl)benzaldehyde

The title compound was prepared in a manner similar to 2-fluoro-4-(6-methyl-4,8-dioxo-1,3,6,2-dioxazaborocan-2-yl)benzaldehyde (intermediate 46b), except that they used (2-fluoro-4-formylphenyl)boronic acid instead of (3-fluoro-4-formylphenyl)boronic acid. ESI–MS 576 [2M+H ₂ O+H] ⁺ .

Example 57 : 2-amino-5-(4-((1R,5S)-3-(2-fluoroethyl)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)-N-(4-hydroxybicyclo [2.2.2]octan–1–yl)nicotinamide

The title compound was prepared in a manner similar to that for 2-amino-5-(4-((1R,5S)-3-(1,3-difluoropropan-2-yl)-3-azabicyclo[3.1.0]hexane-1 -yl)phenyl)-N-((1r,4R)-4-hydroxycyclohexyl)nicotinamide (Example 6), except that the HCl salt and 5-(4-((1R,5S)-3-azabicyclo[ 3.1.0]hexan-1-yl)phenyl)-2-amino-N-(4-hydroxybicyclo[2.2.2]octan-1-yl)nicotinamide (intermediate 34a) instead of 5-(4-((1R, 5S)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)-2-amino-N-((1r,4R)-4-hydroxycyclohexyl)nicotinamide (intermediate 1a) and 1-bromo-2 -fluoroethane instead of 1,3-difluoropropane-2-yltrifluoromethanesulfonate (intermediate 6a). After purification, the title compound was obtained as the TFA salt. ^1H NMR (400 MHz, DMSO–d6) δ 10.00 (bs, 1H), 8.38 (d, 1H), 8.04 (d, 1H), 7.85 (s, 1H), 7. 77 (d, 2H), 7.35 (d, 2H), 7.08 (s, 2H), 4.83 (td, 2H), 4.74 (bs, 1H), 4.08 (bs, 1H ), 3.89–3.57 (m, 4H), 3.18 (d, 1H), 2.24 (bs, 1H), 2.13–1.97 (m, 6H), 1.74– 1.53 (m, 6H), 1.48 (bs, 1H), 1.17 (bs, 1H). (UPLC–MS) t _R 0.53 min; ESI–MS 465 [M+H] ⁺ .

Example 58 : 2-amino-5-(4-((1S,5R)-3-(4,4-difluorocyclohexyl)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)-N-(( 1r,4S)–4–hydroxy–4–methylcyclohexyl)nicotinamide

The title compound was prepared in a manner similar to (1R,5S) -tert -butyl-1-(4-(6-amino-5-(((1r,4R)-4-hydroxycyclohexyl)carbamoyl)pyridin-3-yl )phenyl)-3-azabicyclo[3.1.0]hexane-3-carboxylate (intermediate 1b), except that 2-amino-5-(4-((1S,5R)-3-(4, 4-difluorocyclohexyl)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)nicotinic acid (intermediate 35a) instead of 2-amino-5-(4-((1R,5S)-3-( tert - butoxycarbonyl)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)nicotinic acid (intermediate 1c) and used trans -4-amino-1-methylcyclohexanol instead of trans -4-aminocyclohexanol. The crude product was purified by prepHPLC (method 1a) and passed through an ion exchange cartridge (PL–HCO ₃ ) for desalting to give the title compound. ^1H NMR (400 MHz, DMSO–d6) δ 8.39 (d, 1H), 8.29 (d, 1H), 8.10 (d,1H), 7.59 (d, 2H), 7. 25 (d, 2H), 7.10 (s, 2H), 4.33 (s, 1H), 3.81 (bs, 1H), 3.39 (d, 2H), 2.62–2.56 (m, 1H), 2.00 (bs, 2H), 1.88–1.76 (m, 8H), 1.67–1.53 (m, 4H), 1.50–1.43 (m , 4H), 1.30 (t, 1H), 1.17 (s, 3H), 0.86–0.68 (m, 1H). (UPLC–MS) t _R 0.61 min; ESI–MS 525 [M+H] ⁺ .

Example 59 : 5-(4-((R)-1-(2-oxa-6-azaspiro[3.3]heptan-6-yl)ethyl)phenyl)-2-amino-N-((1r,4R)- 4-hydroxy-4-methylcyclohexyl)nicotinamide

The title compound was prepared in a manner similar to that for 5-(4-((R)-1-(2-oxa-6-azaspiro[3.3]heptan-6-yl)ethyl)phenyl)-2-amino-N-( (1r,4R)-4-hydroxycyclohexyl)nicotinamide (Example 3), except that (6-amino-5-(((1r,4r)-4-hydroxy-4-methylcyclohexyl)carbamoyl)pyridine-3 was used -yl)boronic acid (intermediate 59a) instead of (6-amino-5-(((1r,4r)-4-hydroxycyclohexyl)carbamoyl)pyridin-3-yl)boronic acid (intermediate 3a). ^1H NMR (400 MHz, DMSO–d6) δ 8.38 (s, 1H), 8.25 (bs, 1H), 8.11 (d, 1H) 7.71–7.54 (m, 2H) , 7.49–7.28 (m, 2H), 7.12 (bs, 2H), 4.61 (bs, 4H), 4.31 (s, 1H), 3.89–3.68 (m , 2H), 3.31 (s, 15H), 3.27–2.99(m, 2H), 1.86–1.69 (m, 2H), 1.66–1.52 (m, 2H ), 1.51–1.35 (m, 5H), 1.16 (s, 1H), 1.30–0.95 (m, 2H). (UPLC–MS) t _R 0.47 min; ESI–MS 451 [M+H] ⁺ .

Intermediate 59a : (6-amino-5-(((1r,4r)-4-hydroxy-4-methylcyclohexyl)carbamoyl)pyridin-3-yl)boronic acid

The title compound was prepared in a manner similar to (6-amino-5-(((1r,4r)-4-hydroxycyclohexyl)carbamoyl)pyridin-3-yl)boronic acid (intermediate 3a), except that 2-amino-5-bromo-N-((1r,4r)-4-hydroxy-4-methylcyclohexyl)nicotinamide (intermediate 59b) instead of 2-amino-5-bromo-N-((1r,4r)-4 –hydroxycyclohexyl)nicotinamide (intermediate 3c). (UPLC–MS) t _R 0.33 min; ESI–MS 394 [M+H] ⁺ .

Intermediate 59b : 2-amino-5-bromo-N-((1r,4r)-4-hydroxy-4-methylcyclohexyl)nicotinamide

The title compound was prepared in a manner similar to 2-amino-5-bromo-N-((1r,4r)-4-hydroxycyclohexyl)nicotinamide (intermediate 3c), except that trans -4-amino-1 was used -methylcyclohexanol instead of trans -4-aminocyclohexanol. (UPLC–MS) t _R 0.68 min; ESI–MS 328/330 [M+H] ⁺ .

Example 60 : 2-amino-5-(4-((1R,5S)-3-(but-2-in-1-yl)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)- N–((1r,4R)–4–hydroxycyclohexyl)nicotinamide

The title compound was prepared in a manner similar to that for 2-amino-5-(4-((1R,5S)-3-(1,3-difluoropropan-2-yl)-3-azabicyclo[3.1.0]hexane-1 -yl)phenyl)-N-((1r,4R)-4-hydroxycyclohexyl)nicotinamide (Example 6), except that 1-bromobut-2-yne was used instead of 1,3-difluoropropan-2-yltrifluoromethanesulfonate (intermediate compounds 6a). ^1H NMR (400 MHz, DMSO–d6) δ 8.39 (d, 1H), 8.31 (d, 1H), 8.12 (d, 1H), 7.59 (d, 2H), 7. 21 (d, 2H), 7.12 (s, 2H), 4.58 (d, 1H), 3.77–3.71 (m, 1H), 3.43–3.37 (m, 3H) , 3.22 (d, 1H), 2.92 (d, 1H), 2.80 (d, 1H), 1.93–1.78 (m, 8H), 1.44–1.19 (m , 6H), 0.79 (dd, 1H). (UPLC–MS) t _R 0.54 min; ESI–MS 445 [M+H] ⁺ .

Example 61 : 2-amino-N-cyclohexyl-5-(4-(piperidin-1-yl)phenyl)nicotinamide

The title compound was prepared in a manner similar to 2-amino-N-cyclohexyl-5-(4-(3-(piperidin-1-yl)propoxy)phenyl)nicotinamide (Example 43), except that 4- (piperidin-1-yl)phenylboronic acid instead of 1-(3-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenoxy)propyl)piperidine. ^1H NMR (400 MHz, DMSO–d6) δ 8.33 (d, 1H), 8.33 (s, 1H), 8.08 (d, 1H), 7.51 (d, 2H), 7. 01 (s, 2H), 7.00 (d, 2H), 3.82–3.68 (m, 1H), 3.17 (t, 4H), 1.90–1.78 (m, 2H) , 1.78–1.69 (m, 2H), 1.68–1.49 (m, 7H), 1.38–1.20 (m, 4H), 1.19–1.05 (m, 1H). (HPLC–MS) t _R 1.40 min; APCI–MS 379 [M+H] ⁺ .

Example 62 : 2-amino-N-(3-(hydroxymethyl)bicyclo[1.1.1]pentan-1-yl)-5-(4-((1S,5R)-3-(tetrahydro-2H-pyran-4 –yl)-3-azabicyclo[3.1.0]hexane-1-yl)phenyl)nicotinamide

The title compound was prepared in a manner similar to (1R,5S) -tert -butyl-1-(4-(6-amino-5-(((1r,4R)-4-hydroxycyclohexyl)carbamoyl)pyridin-3-yl )phenyl)-3-azabicyclo[3.1.0]hexane-3-carboxylate (intermediate 1b), except that 2-amino-5-(4-((1S,5R)-3-(tetrahydro- 2H-pyran-4-yl)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)nicotinic acid (intermediate 28a) instead of 2-amino-5-(4-((1R,5S)-3 -( tert -butoxycarbonyl)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)nicotinic acid (intermediate 1c) and used trans -4-amino-1-methylcyclohexanol instead of trans -4-aminocyclohexanol. The crude product was first purified by prepHPLC (Method 1a) followed by reverse phase flash chromatography (Method 3a) to obtain the title compound as a colorless solid. ^1H NMR (400 MHz, DMSO–d6) δ 8.92 (d, 1H), 8.46 (s, 1H), 8.22 (s, 1H), 7.61 (d, 2H), 7. 25 (d, 2H), 7.18 (s, 2H), 4.15–4.09 (m, 1H), 3.88–3.82 (m, 2H), 3.42–3.36 ( m, 3H), 3.10 (bs, 1H), 2.65–2.34 (m, 12H), 1.85–1.75 (m, 3H), 1.45–1.31 (m, 6H), 0.77 (bs, 1H). (UPLC–MS) t _R 0.49 min; ESI–MS 491 [M+H] ⁺ .

Example 63 : 2-amino-N-(4-hydroxybicyclo[2.2.2]octan-1-yl)-5-(4-((1S,5R)-3-(oxetan-3-yl)-3-azabicyclo [3.1.0]hexan–1–yl)phenyl)nicotinamide

The title compound was prepared in a manner similar to that for 2-amino-N-((1r,4R)-4-hydroxycyclohexyl)-5-(4-((1R,5S)-3-(1-(methylsulfonyl)piperidine-4 -yl)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)nicotinamide (Example 1), except that the HCl salt and 5-(4-((1S,5R)-3-azabicyclo [3.1.0]hexan-1-yl)phenyl)-2-amino-N-(4-hydroxybicyclo[2.2.2]octan-1-yl)nicotinamide (intermediate 40a) instead of 5-(4-((1R ,5S)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)-2-amino-N-((1r,4R)-4-hydroxycyclohexyl)nicotinamide (intermediate 1a) and oxetane-3- it instead of 1-(methylsulfonyl)piperidin-4-one. ^1H NMR (400 MHz, DMSO–d6) δ 8.34 (d, 1H), 7.98 (d, 1H), 7.80 (s, 1H), 7.57 (d, 2H), 7. 22 (d, 2H), 6.92 (s, 2H), 4.61–4.56 (m, 2H), 4.52–4.48 (m, 2H), 4.32 (s, 1H) , 3.80–3.74 (m, 1H), 3.03 (d, 1H), 2.57–2.51 (m, 3H), 2.14–1.98 (m, 6H), 1 .88–1.85 (m, 1H), 1.71–1.55 (m, 6H), 1.45–1.35 (m, 1H), 0.83–0.80 (m, 1H) . (UPLC–MS) t _R 0.51 min; ESI–MS 475 [M+H] ⁺ .

Example 64 : 2-amino-N-(4-hydroxybicyclo[2.2.2]octan-1-yl)-5-(4-((1R,5S)-3-(oxetan-3-yl)-3-azabicyclo [3.1.0]hexan–1–yl)phenyl)nicotinamide

The title compound was prepared in a manner similar to that for 2-amino-N-((1r,4R)-4-hydroxycyclohexyl)-5-(4-((1R,5S)-3-(1-(methylsulfonyl)piperidine-4 -yl)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)nicotinamide (Example 1), except that the HCl salt and 5-(4-((1R,5S)-3-azabicyclo [3.1.0]hexan-1-yl)phenyl)-2-amino-N-(4-hydroxybicyclo[2.2.2]octan-1-yl)nicotinamide (intermediate 34a) instead of 5-(4-((1R ,5S)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)-2-amino-N-((1r,4R)-4-hydroxycyclohexyl)nicotinamide (intermediate 1a) and oxetane-3- it instead of 1-(methylsulfonyl)piperidin-4-one. ^1H NMR (400 MHz, DMSO–d6) δ 8.34 (d, 1H), 7.98 (d, 1H), 7.80 (s, 1H), 7.57 (d, 2H), 7. 22 (d, 2H), 6.92 (s, 2H), 4.61–4.56 (m, 2H), 4.52–4.48 (m, 2H), 4.31 (s, 1H) , 3.81–3.76 (m, 1H), 3.03 (d, 1H), 2.57–2.51 (m, 3H), 2.07–2.03 (m, 6H), 1 .88–1.85 (m, 1H), 1.65–1.61 (m, 6H), 1.45–1.35 (m, 1H), 0.83–0.80 (m, 1H) . (UPLC–MS) t _R 0.55 min; ESI–MS 475 [M+H] ⁺ .

Example 65 : 2-amino-N-cyclohexyl-5-(4-((2-(4-methylpiperazin-1-yl)ethyl)carbamoyl)phenyl)nicotinamide

The title compound was prepared in a manner similar to (1R,5S) –tert -butyl–1–(4–(6–amino–5–(((1r,4R)–4–hydroxycyclohexyl)carbamoyl)pyridin–3–yl )phenyl)-3-azabicyclo[3.1.0]hexane-3-carboxylate (intermediate 1b), except that 4-(6-amino-5-(cyclohexylcarbamoyl)pyridin-3-yl)benzoic acid was used ( intermediate 65a) instead of 2-amino-5-(4-((1R,5S)-3-( tert -butoxycarbonyl)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)nicotinic acid (intermediate 1c) and used 2-(4-methylpiperazin-1-yl)ethanamine instead of trans -4-aminocyclohexanol. The crude product was purified by normal phase chromatography (method 2c) to give the title compound as an off-white solid. ^1H NMR (400 MHz, DMSO–d6) δ 8.50 (d, 1H), 8.42 (t, 1H), 8.37 (d, 1H), 8.23 (d, 1H), 7. 91 (d, 2H), 7.80 (d, 2H), 7.25 (s, 2H), 3.82–3.65 (m, 1H), 3.40 (q, 2H), 3.32 –3.13 (m, 3H), 2.99–2.83 (m, 1H), 2.43–2.31 (m, 6H), 2.17 (s, 3H), 1.90–1 .80 (m, 2H), 1.80–1.70 (m, 2H), 1.70–1.55 (m, 1H), 1.40–1.25 (m, 4H), 1.20 –1.10 (m, 1H). (HPLC–MS) t _R 1.28 min; APCI–MS 465 [M+H] ⁺ .

Intermediate 65a : 4-(6-amino-5-(cyclohexylcarbamoyl)pyridin-3-yl)benzoic acid

The title compound was prepared in a manner similar to 2-amino-N-cyclohexyl-5-(4-(3-(piperidin-1-yl)propoxy)phenyl)nicotinamide (Example 43), except that 4- (4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzoic acid instead of 1-(3-(4-(4,4,5,5-tetramethyl-1,3,2 –dioxaborolan-2-yl)phenoxy)propyl)piperidine. The crude product was triturated with 2-propanol to give a brownish solid, which was purified by prepHPLC (method 1a). The pure fractions were collected and concentrated under reduced pressure, then increased in basicity with Na ₂ CO ₃ , after which the title compound precipitated as a colorless solid, which was filtered and dried under reduced pressure. (HPLC–MS) t _R 1.52 min; ESI–MS 340 [M+H] ⁺ .

Example 66 : 2-amino-5-(4-((1S,5R)-3-(1,3-difluoropropan-2-yl)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)- N–((1r,4S)–4–hydroxycyclohexyl)nicotinamide

The title compound was prepared in a manner similar to that for 2-amino-5-(4-((1R,5S)-3-(1,3-difluoropropan-2-yl)-3-azabicyclo[3.1.0]hexane-1 -yl)phenyl)-N-((1r,4R)-4-hydroxycyclohexyl)nicotinamide (Example 5), except that 1,3-difluoropropan-2-yltrifluoromethanesulfonate (intermediate 6a) was used instead of 4-(3 –bromopropyl)morpholine. ^1H NMR (400 MHz, DMSO–d6) δ 8.39 (d, 1H), 8.31 (d, 1H), 8.12 (d, 1H), 7.59 (d, 2H), 7. 24 (d, 2H), 7.12 (s, 2H), 4.68 (t, 2H), 4.57 (dd, 3H), 3.81–3.65 (m, 1H), 3.46 –3.35 (m, 2H), 3.08 (d, 1H), 2.93 (d, 1H), 2.83 (dd, 1H), 2.05–1.76 (m, 5H), 1.53–1.16 (m, 6H), 0.78 (dd, 1H). (UPLC–MS) t _R 0.68 min; ESI–MS 471 [M+H] ⁺ .

Example 67 : 2-amino-N-((1r,4S)-4-hydroxycyclohexyl)-5-(4-((1S,5R)-3-(2,2,2-trifluoroethyl)-3-azabicyclo[3.1 .0]hexan–1–yl)phenyl)nicotinamide

The title compound was prepared in a manner similar to that for 2-amino-5-(4-((1R,5S)-3-(1,3-difluoropropan-2-yl)-3-azabicyclo[3.1.0]hexane-1 -yl)phenyl)-N-((1r,4R)-4-hydroxycyclohexyl)nicotinamide (Example 5), except that 2,2,2-trifluoroethyltrifluoromethanesulfonate was used instead of 4-(3-bromopropyl)morpholine. ^1H NMR (400 MHz, DMSO–d6) δ 8.46–8.31 (m, 2H), 8.18 (bs, 1H), 7.59 (d, 2H), 7.29 (bs, 1H ), 7.23 (d, 2H), 3.83–3.54 (m, 1H), 3.51–3.24 (m, 5H), 3.11 (d, 1H), 2.92 ( d, 1H), 2.86 (m, 1H), 1.92–1.77 (m, 5H), 1.29–1.21 (m, 1H), 1.42–1.18 (m, 4H), 1.14 (bs, 1H), 0.85–0.79 (m, 1H). (UPLC–MS) t _R 1.04 min; ESI–MS 475 [M+H] ⁺ .

Example 68 : 5-(4-((R)-1-(2-oxa-6-azaspiro[3.3]heptan-6-yl)ethyl)phenyl)-2-amino-N-((1r,3R)- 3-hydroxyadamantan-1-yl)nicotinamide

The title compound was prepared in a manner similar to that for 5-(4-((R)-1-(2-oxa-6-azaspiro[3.3]heptan-6-yl)ethyl)phenyl)-2-amino-N-( (1r,4R)-4-hydroxycyclohexyl)nicotinamide (Example 3), except that (6-amino-5-(((1r,3r)-3-hydroxyadamantan-1-yl)carbamoyl)pyridine-3 was used -yl)boronic acid (intermediate 68a) instead of (6-amino-5-(((1r,4r)-4-hydroxycyclohexyl)carbamoyl)pyridin-3-yl)boronic acid (intermediate 3a). ^1H NMR (400 MHz, DMSO–d6) δ 8.65 (bs, 1H), 8.49 (s, 1H), 8.35 (s, 1H), 7.87 (d, 2H), 7. 59 (d, 2H), 4.73 (dd, 2H), 4.65–4.54 (m, 4H), 4.45–4.39 (m, 1H), 4.16–4.10 ( m, 1H), 3.98 (s, 1H), 3.90 (bs, 1H), 2.21 (s, 2H), 2.04–1.94 (m, 6H), 1.60 (bs , 2H), 1.49–1.43 (m, 3H). (UPLC–MS) t _R 0.56 min; ESI–MS 489 [M+H] ⁺ .

Intermediate 68a : 6-amino-5-(((1r,3r)-3-hydroxyadamantan-1-yl)carbamoyl)pyridin-3-yl)boronic acid

The title compound was prepared in a manner similar to (6-amino-5-(((1r,4r)-4-hydroxycyclohexyl)carbamoyl)pyridin-3-yl)boronic acid (intermediate 3a), except that 2-amino-5-bromo-N-((1r,3r)-3-hydroxyadamantan-1-yl)nicotinamide (intermediate 68b) instead of 2-amino-5-bromo-N-((1r,4r)-4 –hydroxycyclohexyl)nicotinamide (intermediate 3c). (UPLC–MS) t _R 0.47 min; ESI–MS 332 [M+H] ⁺ .

Intermediate 68b : 2-amino-5-bromo-N-((1r,3r)-3-hydroxyadamantan-1-yl)nicotinamide

The title compound was prepared in a manner similar to 2-amino-5-bromo-N-((1r,4r)-4-hydroxycyclohexyl)nicotinamide (intermediate 3c), except that 3-amino-1-adamantanol was used instead of trans -4-aminocyclohexanol. (UPLC–MS) t _R 0.82 min; ESI–MS 366/368 [M+H] ⁺ .

Example 69 : 2-amino-N-cyclohexyl-5-(4-(morpholine-4-carbonyl)phenyl)nicotinamide

The title compound was prepared in a manner similar to 2-amino-N-cyclohexyl-5-(4-(3-(piperidin-1-yl)propoxy)phenyl)nicotinamide (Example 43), except that 4- (morpholine-4-carbonyl)phenylboronic acid instead of 1-(3-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenoxy)propyl)piperidine. ^1H NMR (400 MHz, DMSO–d6) δ 8.47 (d, 1H), 8.42 (s, 1H), 8.30 (d, 1H), 7.78 (d, 2H), 7. 51 (d, 2H), 7.49 (s, 2H), 3.85–3.70 (m, 1H), 3.70–3.50 (m, 8H), 1.90–1.80 ( m, 2H), 1.80–1.70 (m, 2H), 1.70–1.55 (m, 1H), 1.35–1.29 (m, 4H), 1.28–1, 05 (m, 1H). (HPLC–MS) t _R 1.51 min; APCI–MS 409 [M+H] ⁺ .

Example 70 : 2-amino-N-((1r,4R)-4-hydroxycyclohexyl)-5-(4-((1R,5S)-3-(oxetan-3-ylmethyl)-3-azabicyclo[3.1.0 ]hexan–1–yl)phenyl)nicotinamide

The title compound was prepared in a manner similar to that for 2-amino-5-(4-((1R,5S)-3-(1,3-difluoropropan-2-yl)-3-azabicyclo[3.1.0]hexane-1 -yl)phenyl)-N-((1r,4R)-4-hydroxycyclohexyl)nicotinamide (Example 6), except that oxetan-3-ylmethyltrifluoromethanesulfonate (intermediate 70a) was used instead of 1,3-difluoropropane-2- yltrifluoromethanesulfonate (intermediate 6a) and the reaction mixture was stirred at 60°C for 3 hours. ^1H NMR (400 MHz, DMSO–d6) δ 8.38 (d, 1H), 8.30 (d, 1H), 8 .11 (d, 1H), 7.58 (d, 2H), 7.21 (d, 2H), 7.12 (s, 2H), 4.66 (dt, 2H), 4.58 (d, 1H), 4.28 (t, 2H), 3.90–3.66 (m, 1H), 3.43–3.35 (m, 1H), 3.25 (dd, 1H), 3.19 –3.09 (m, 1H), 2.97 (d, 1H), 2.79 (d, 2H), 2.62–2.51 (m, 2H), 1.98–1.75 (m , 5H), 1.48–1.13 (m, 5H), 0.75 (dd, 1H). (UPLC–MS) t _R 0.46 min; ESI–MS 463 [M+H] ⁺ .

Intermediate 70a : Oxetan-3-ylmethyltrifluoromethanesulfonate

The title compound was prepared in a manner similar to 1,3-difluoropropan-2-yltrifluoromethanesulfonate (intermediate 6a), except that oxetan-3-ylmethanol was used instead of 1,3-difluoropropan-2-ol and was prepared in in the form of crude oil, which was used without further purification.

Example 71 : 2-amino-N-((1s,4R)-4-hydroxycyclohexyl)-5-(4-((1S,5R)-3-isopropyl-3-azabicyclo[3.1.0]hexan-1-yl )phenyl)nicotinamide

The title compound was prepared in a manner similar to (1R,5S) -tert -butyl-1-(4-(6-amino-5-(((1r,4R)-4-hydroxycyclohexyl)carbamoyl)pyridin-3-yl )phenyl)-3-azabicyclo[3.1.0]hexane-3-carboxylate (intermediate 1b), except that 2-amino-5-(4-((1S,5R)-3-isopropyl-3 -azabicyclo[3.1.0]hexan-1-yl)phenyl)nicotinic acid (intermediate 71a) instead of 2-amino-5-(4-((1R,5S)-3-( tert -butoxycarbonyl)-3-azabicyclo [3.1.0]hexan-1-yl)phenyl)nicotinic acid (intermediate 1c) and used cis -4-aminocyclohexanol hydrochloride instead of trans -4-aminocyclohexanol. The crude product was purified by prepHPLC (Method 1a) to obtain the title compound as a yellow solid. ^1H NMR (400 MHz, DMSO–d6) δ 8.49–8.34 (m, 2H), 8.17 (s, 1H), 7.67 (s, 2H), 7.36 (s, 2H ), 7.16 (s, 2H), 4.43 (d, 1H), 3.81 (s, 2H), 3.81–3.59 (m, 4H), 1.88–1.62 ( m, 5H), 1.61–1.44 (m, 5H), 1.35 (s, 6H), 1.16 (s, 2H). (UPLC–MS) t _R 0.52 min; ESI–MS 435 [M+H] ⁺ .

Intermediate 71a : 2-amino-5-(4-((1S,5R)-3-isopropyl-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)nicotinic acid

The title compound was prepared in a manner similar to that for 2-amino-5-(4-((1R,5S)-3-( tert -butoxycarbonyl)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)nicotinic acid (intermediate 1c), except that methyl-2-amino-5-(4-((1S,5R)-3-isopropyl-3-azabicyclo[3.1.0]hexan-1-yl)phenyl was used )nicotinate (intermediate 71b) instead of (1R,5S) -tert -butyl-1-(4-(6-amino-5-(methoxycarbonyl)pyridin-3-yl)phenyl)-3-azabicyclo[3.1.0] hexane-3-carboxylate (intermediate 1d). (UPLC–MS) t _R 0.40 min; ESI–MS 338 [M+H] ⁺ .

Intermediate 71b : Methyl-2-amino-5-(4-((1S,5R)-3-isopropyl-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)nicotinate

The title compound was prepared in a manner similar to that for methyl-2-amino-5-(4-((1S,5R)-3-(2-methoxyethyl)-3-azabicyclo[3.1.0]hexane-1-yl)phenyl )nicotinate (intermediate 7b), except that 2-iodopropane was used instead of 1-bromo-2-methoxyethane. (UPLC–MS) t _R 0.65 min; ESI–MS 352 [M+H] ⁺ .

Example 72 : 2-amino-N-((1r,4R)-4-hydroxy-4-methylcyclohexyl)-5-(4-(((R)-2-methylpyrrolidin-1-yl)methyl)phenyl)nicotinamide

(R)-2-methylpyrrolidine (29.0 mg, 0.340 mmol) and K ₂ CO ₃ ( 78 mg, 0.567 mmol) at room temperature. under a nitrogen atmosphere and the resulting white suspension was stirred at 60°C for 2 hours. Water (1 ml) was added at room temperature. followed by the addition of 2-amino-5-bromo-N-((1r,4r)-4-hydroxy-4-methylcyclohexyl)nicotinamide (intermediate 59b, 150 mg, 0.283 mmol) in acetonitrile (2 ml). PdCl ₂ (dppf) (10.37 mg, 0.014 mmol) was then added and the reaction mixture was stirred at 90°C for 60 minutes. After filtration through a pad of celite and concentration under reduced pressure, the crude product was diluted with sat. aq. a solution of NaHCO ₃ and EtOAc. After phase separation, the aqueous layer was extracted with EtOAc and the combined organic layers were washed with brine, dried over MgSO ₄ , filtered and concentrated under reduced pressure. The resulting crude product was first purified by prepHPLC (Method 1a) then by normal phase chromatography (Method 2b) to obtain the title compound as a colorless solid. ^1H NMR (400 MHz, DMSO–d6) δ 9.75 (d, 1H), 8.52 (t, 1H), 8.41 (d, 1H), 8.27 (d, 1H), 7. 85 (d, 2H), 7.66 (d, 2H), 7.38 (bs, 2H), 4.67–4.53 (m, 1H), 4.25 (dd, 1H), 3.85 (s, 1H), 3.58–3.49 (m, 1H), 3.37–3.18 (m, 3H), 2.29 (m, 1H), 2.06–1.88 (m , 2H), 1.83 (s, 2H), 1.65 (dd, 2H), 1.51 (d, 4H), 1.39 (dd, 3H), 1.22 (d, 3H). (UPLC–MS) t _R 0.47 min; ESI–MS 423 [M+H] ⁺ .

Example 73 : 2-amino-5-(4-((1S,5R)-3-(2-fluoroethyl)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)-N-(4-hydroxybicyclo [2.2.2]octan–1–yl)nicotinamide

The title compound was prepared in a manner similar to that for 2-amino-5-(4-((1R,5S)-3-(1,3-difluoropropan-2-yl)-3-azabicyclo[3.1.0]hexane-1 -yl)phenyl)-N-((1r,4R)-4-hydroxycyclohexyl)nicotinamide (Example 5), except that the HCl salt and 5-(4-((1S,5R)-3-azabicyclo[ 3.1.0]hexan-1-yl)phenyl)-2-amino-N-(4-hydroxybicyclo[2.2.2]octan-1-yl)nicotinamide (intermediate 40a) instead of 5-(4-((1S, 5R)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)-2-amino-N-((1r,4R)-4-hydroxycyclohexyl)nicotinamide (intermediate 5a) and 1-bromo-2 –fluoroethane instead of 4-(3-bromopropyl)morpholine, and the reaction mixture was stirred at 60°C for 3 hours. ^1H NMR (400 MHz, DMSO–d6) δ 8.35 (d, 1H), 7.98 ( d, 1H), 7.79 (s, 1H), 7.57 (d, 2H), 7.22 (d, 2H), 6.92 (s, 2H), 4.55 (td, 2H), 4.31 (s, 1H), 3.38 (d, 1H), 3.10 (d, 1H), 2.84 (t, 1H), 2.76 (t, 1H), 2.64 (d , 1H), 2.54–2.51 (m, 1H), 2.13–1.96 (m, 6H), 1.83 (dd, 1H), 1.73–1.54 (m, 6H ), 1.34 (t, 1H), 0.78 (dd, 1H). (UPLC–MS) t _R 0.51 min; ESI–MS 465 [M+H] ⁺ .

Example 74 : 2-amino-5-(2,3-difluoro-4-(((R)-2-methylpyrrolidin-1-yl)methyl)phenyl)-N-((1r,4R)-4-hydroxy- 4-methylcyclohexyl)nicotinamide

The title compound was prepared in a manner similar to that for 2-amino-5-(3-fluoro-4-(((R)-2-methylpyrrolidin-1-yl)methyl)phenyl)-N-((1r,4R)- 4-hydroxy-4-methylcyclohexyl)nicotinamide (example 46), except that (R)-2-(2,3-difluoro-4-((2-methylpyrrolidin-1-yl)methyl)phenyl)- 6-methyl-1,3,6,2-dioxazaborocane-4,8-dione (intermediate 74a) instead of (R)-2-(3-fluoro-4-((2-methylpyrrolidin-1-yl)methyl) phenyl)-6-methyl-1,3,6,2-dioxazaborocane-4,8-dione (intermediate 46a). (UPLC–MS) t _R 0.76 min; ESI–MS 459 [M+H] ⁺ .

Intermediate 74a : (R)-2-(2,3-difluoro-4-((2-methylpyrrolidin-1-yl)methyl)phenyl)-6-methyl-1,3,6,2-dioxazaborocane-4, 8-dion

The title compound was prepared in a manner similar to that for (R)-2-(3-fluoro-4-((2-methylpyrrolidin-1-yl)methyl)phenyl)-6-methyl-1,3,6,2-dioxazaborocane –4,8-dione (intermediate 46a), except that 2,3-difluoro-4-(6-methyl-4,8-dioxo-1,3,6,2-dioxazaborocane-2-yl) was used )benzaldehyde (intermediate 74b) instead of 2-fluoro-4-(6-methyl-4,8-dioxo-1,3,6,2-dioxazaborocan-2-yl)benzaldehyde (intermediate 46b). ESI–MS 367 [M+H] ⁺ .

Intermediate 74b : 2,3-difluoro-4-(6-methyl-4,8-dioxo-1,3,6,2-dioxazaborocane-2-yl)benzaldehyde

The title compound was prepared in a manner similar to 2-fluoro-4-(6-methyl-4,8-dioxo-1,3,6,2-dioxazaborocan-2-yl)benzaldehyde (intermediate 46b), except that they used (2,3-difluoro-4-formylphenyl)boronic acid instead of (3-fluoro-4-formylphenyl)boronic acid. ESI–MS 595 [2M+H] ⁺ .

Example 75 : 2-amino-N-((1r,4R)-4-hydroxycyclohexyl)-5-(4-((1R,5S)-3-(2-methoxyethyl)-3-azabicyclo[3.1.0]hexane –1-yl)phenyl)nicotinamide

The title compound was prepared in a manner similar to that for 2-amino-5-(4-((1R,5S)-3-(1,3-difluoropropan-2-yl)-3-azabicyclo[3.1.0]hexane-1 -yl)phenyl)-N-((1r,4R)-4-hydroxycyclohexyl)nicotinamide (Example 6), except that 1-bromo-2-methoxyethane was used instead of 1,3-difluoropropan-2-yltrifluoromethanesulfonate (intermediate compound 6a) and the reaction mixture was stirred at 60°C for 3 hours. After purification, the title compound was obtained as the TFA salt. ^1H NMR (400 MHz, DMSO–d6) δ 9.70 (bs, 1H), 8.41 (s, 1H), 8.31 (d, 1H), 8.12 (d, 1H), 7, 66–7.60 (m, 2H), 7.30 (bs, 2H), 7.14 (s, 2H), 4.59 (d, 1H), 3.81–3.38 (m, 8H) , 3.31 (s, 3H), 2.75–2.25 (m, 4H), 1.86 (t, 4H), 1.50–1.21 (m, 6H). (UPLC–MS) t _R 0.48 min; ESI–MS 451 [M+H] ⁺ .

Example 76 : 2-amino-5-(4-((1S,5R)-3-(2,2-difluoropropyl)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)-N-(( 1r,4S)–4–hydroxycyclohexyl)nicotinamide

The title compound was prepared in a manner similar to that for 2-amino-5-(4-((1R,5S)-3-(1,3-difluoropropan-2-yl)-3-azabicyclo[3.1.0]hexane-1 -yl)phenyl)-N-((1r,4R)-4-hydroxycyclohexyl)nicotinamide (Example 5), except that 2,2-difluoropropyltrifluoromethanesulfonate was used instead of 4-(3-bromopropyl)morpholine. ^1H NMR (400 MHz, DMSO–d6) δ 8.39 (d, 1H), 8.33 (d, 1H), 8.14 (d, 1H), 7.60 (d, 2H), 7. 32–7.09 (m, 4H), 4.59 (bs, 1H), 3.74 (bs, 1H), 3.11 (d, 1H), 2.98–2.63 (m, 5H) , 1.89–1.83 (m, 5H), 1.63 (t, 3H), 1.42–1.25 (m, 6H), 0.81 (bs, 1H). (UPLC–MS) t _R 0.82 min; ESI–MS 471 [M+H] ⁺ .

Example 77 : 2-amino-N-((1r,4R)-4-hydroxycyclohexyl)-5-(4-((1R,5S)-3-(2,2,2-trifluoroethyl)-3-azabicyclo[3.1 .0]hexan–1–yl)phenyl)nicotinamide

The title compound was prepared in a manner similar to that for 2-amino-5-(4-((1R,5S)-3-(1,3-difluoropropan-2-yl)-3-azabicyclo[3.1.0]hexane-1 -yl)phenyl)-N-((1r,4R)-4-hydroxycyclohexyl)nicotinamide (Example 6), except that 2,2,2-trifluoroethyltrifluoromethanesulfonate was used instead of 1,3-difluoropropan-2-yltrifluoromethanesulfonate (intermediate compound 6a) and the reaction mixture was stirred at 60°C for 1 h. ¹ H NMR (400 MHz, DMSO–d6) δ 8.39 (d, 1H), 8.32 (d, 1H), 8.13 ( d, 1H), 7.60 (d, 2H), 7.24 (d, 2H), 7.16 (s, 2H), 4.58 (s, 1H), 3.73 (dt, 1H), 3.41 (m, 4H), 3.13 (d, 1H), 2.93 (d, 1H), 2.84 (dd, 1H), 1.92–1.77 (m, 5H), 1 .51–1.15 (m, 5H), 0.84 (dd, 1H). (UPLC–MS) t _R 1.03 min; ESI–MS 475 [M+H] ⁺ .

Example 78 : 2-amino-N-((1r,4S)-4-hydroxycyclohexyl-4- d )-5-(4-((1S,5R)-3-isopropyl-3-azabicyclo[3.1.0]hexane –1-yl)phenyl)nicotinamide

The title compound was prepared in a manner similar to (1R,5S) -tert -butyl-1-(4-(6-amino-5-(((1r,4R)-4-hydroxycyclohexyl)carbamoyl)pyridin-3-yl )phenyl)-3-azabicyclo[3.1.0]hexane-3-carboxylate (intermediate 1b), except that 2-amino-5-(4-((1S,5R)-3-isopropyl-3 -azabicyclo[3.1.0]hexan-1-yl)phenyl)nicotinic acid (intermediate 71a) instead of 2-amino-5-(4-((1R,5S)-3-( tert -butoxycarbonyl)-3-azabicyclo [3.1.0]hexan-1-yl)phenyl)nicotinic acid (intermediate 1c) and used trans -4-aminocyclohexan-1- d -1-ol hydrochloride (intermediate 78a) instead of trans -4-aminocyclohexanol. The crude product was first purified by prepHPLC (Method 1a) then by reverse phase flash chromatography (Method 3a) to obtain the title compound as a colorless solid. ^1H NMR (400 MHz, DMSO–d6) δ 8.38 (d, 1H), 8.31 (d, 1H), 8.11 (d, 1H), 7.58 (d, 2H), 7. 22 (d, 2H), 7.12 (s, 2H), 4.56 (s, 1H), 3.73 (dt, 1H), 3.37 (s, 1H), 3.06 (d, 1H ), 2.57 (d, 1H), 2.49–2.41 (m, 2H), 1.93–1.73 (m, 5H), 1.31 (m, 5H), 1.04 ( dd, 6H), 0.74 (dd, 1H). (UPLC–MS) t _R 0.50 min; ESI–MS 436 [M+H] ⁺ .

Intermediate 78a : trans -4-aminocyclohexane-1- d -1-ol hydrochloride

Deuterated aminocyclohexyl starting materials can be prepared using methods similar to those known in the art (eg, Quirante, J. et al., J. Org. Chem. 67(7): 2323–2328 (2002)). For example, tert -butylbenzyl(4-oxocyclohexyl)carbamate can be reduced with NaBD ₄ to produce deuterated tert -butylbenzyl((1r,4r)-4-hydroxycyclohexyl)carbamate, which, after removal of the tert -butyloxycarbonyl and benzyl protecting groups, can be used in the synthesis methods described herein.

Example 79 : TFA salt of 2-amino-N-((1s,4s)-4-hydroxycyclohexyl)-5-(4-(4-methylpiperazine-1-carbonyl)phenyl)nicotinamide

The title compound was prepared in a manner similar to 2-amino-N-cyclohexyl-5-(4-(3-(piperidin-1-yl)propoxy)phenyl)nicotinamide (Example 43), except that 2- amino-5-bromo-N-((1s,4s)-4-hydroxycyclohexyl)nicotinamide (intermediate 79a) instead of 2-amino-5-bromo-N-cyclohexylnicotinamide (intermediate 43a) and used (4-methylpiperazine-1 –yl)(4–(4,4,5,5–tetramethyl–1,3,2–dioxaborolan–2–yl)phenyl)methanone instead of 1–(3–(4–(4,4,5,5– tetramethyl-1,3,2-dioxaborolan-2-yl)phenoxy)propyl)piperidine. The reaction was carried out in a microwave reactor (irradiation time 15 min at 80°C). The title compound was obtained as the TFA salt. ^1H NMR (400 MHz, DMSO–d6) δ 10.01 (s, 2H), 8.56 (d, 1H), 8.51 (d, 1H), 8.43 (d, 1H), 7, 85 (d, 2H), 7.57 (d, 2H), 3.86–3.55 (m, 3H), 3.55–3.20 (m, 4H), 3.40–3.15 ( m, 3H), 2.85 (s, 3H), 1.85–1.70 (m, 2H), 1.70–1.65 (m, 2H), 1.65–1.40 (m, 4H). (HPLC–MS) t _R 0.35 min; ESI–MS 438 [M+H] ⁺ .

Intermediate 79a : 2-amino-5-bromo-N-((1s,4s)-4-hydroxycyclohexyl)nicotinamide

The title compound was prepared in a manner similar to 2-amino-5-bromo-N-((1r,4r)-4-hydroxycyclohexyl)nicotinamide (intermediate 3c), except that cis -4-aminocyclohexanol was used instead of trans –4-aminocyclohexanol. (UPLC–MS) t _R 0.53 min; ESI–MS 314/316 [M+H] ⁺ .

Example 80 : 2-amino-5-(4-((1R,5S)-3-(4,4-difluorocyclohexyl)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)-N-(4 –hydroxybicyclo[2.2.2]octan-1-yl)nicotinamide

The title compound was prepared in a manner similar to that for 2-amino-N-((1r,4R)-4-hydroxycyclohexyl)-5-(4-((1R,5S)-3-(1-(methylsulfonyl)piperidine-4 -yl)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)nicotinamide (Example 1), except that the HCl salt and 5-(4-((1R,5S)-3-azabicyclo [3.1.0]hexan-1-yl)phenyl)-2-amino-N-(4-hydroxybicyclo[2.2.2]octan-1-yl)nicotinamide (intermediate 34a) instead of 5-(4-((1R ,5S)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)-2-amino-N-((1r,4R)-4-hydroxycyclohexyl)nicotinamide (intermediate 1a) and used 4,4- difluorocyclohexanone instead of 1-(methylsulfonyl)piperidin-4-one. ^1H NMR (400 MHz, DMSO–d6) δ 8.32 (d, 1H), 7.95 (d, 1H), 7.76 (s, 1H), 7.55 (d, 2H), 7. 21 (d, 2H), 6.88 (s, 2H), 4.28 (s, 1H), 3.36 (s, 1H), 3.07 (s, 1H), 2.55 (s, 1H ), 2.45 (s, 1H), 2.32 (d, 1H), 2.30–1.95 (m, 8H), 1.77 (d, 5H), 1.70–1.32 ( m, 8H), 1.27 (t, 1H), 0.75 (s, 1H). (UPLC–MS) t _R 0.63 min; ESI–MS 537 [M+H] ⁺ .

Example 81 : TFA salt and 2-amino-5-(2-chloro-4-(((R)-2-methylpyrrolidin-1-yl)methyl)phenyl)-N-((1r,4R)-4-hydroxy –4-methylcyclohexyl)nicotinamide

The title compound was prepared in a manner similar to that for 2-amino-5-(3-fluoro-4-(((R)-2-methylpyrrolidin-1-yl)methyl)phenyl)-N-((1r,4R)- 4-hydroxy-4-methylcyclohexyl)nicotinamide (example 46), except that (R)-2-(2-chloro-4-((2-methylpyrrolidin-1-yl)methyl)phenyl)-6- methyl-1,3,6,2-dioxazaborocane-4,8-dione (intermediate 81a) instead of (R)-2-(3-fluoro-4-((2-methylpyrrolidin-1-yl)methyl)phenyl) –6-methyl-1,3,6,2-dioxazaborocane-4,8-dione (intermediate 46a). UPLC–HRMS t _R 3.45 min, 457.24/459.24 [M+H] ⁺ .

Intermediate 81a : (R)-2-(2-chloro-4-((2-methylpyrrolidin-1-yl)methyl)phenyl)-6-methyl-1,3,6,2-dioxazaborocane-4,8- dione

The title compound was prepared in a manner similar to that for (R)-2-(3-fluoro-4-((2-methylpyrrolidin-1-yl)methyl)phenyl)-6-methyl-1,3,6,2-dioxazaborocane –4,8-dione (intermediate 46a), except that 3-chloro-4-(6-methyl-4,8-dioxo-1,3,6,2-dioxazaborocan-2-yl)benzaldehyde was used (intermediate 81b) instead of 2-fluoro-4-(6-methyl-4,8-dioxo-1,3,6,2-dioxazaborocan-2-yl)benzaldehyde (intermediate 46b). ESI–MS 365/367 [M+H] ⁺ .

Intermediate 81b : 3-chloro-4-(6-methyl-4,8-dioxo-1,3,6,2-dioxazaborocan-2-yl)benzaldehyde

The title compound was prepared in a manner similar to 2-fluoro-4-(6-methyl-4,8-dioxo-1,3,6,2-dioxazaborocan-2-yl)benzaldehyde (intermediate 46b), except that they used (2-chloro-4-formylphenyl)boronic acid instead of (3-fluoro-4-formylphenyl)boronic acid. ESI–MS 313/315 [M+H ₂ O+H] ⁺ .

Example 82 : 2-amino-5-(4-((1R,5S)-3-(4,4-difluorocyclohexyl)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)-N-(( 1r,4R)–4–hydroxy–4–methylcyclohexyl)nicotinamide

The title compound was prepared in a manner similar to (1R,5S) -tert -butyl-1-(4-(6-amino-5-(((1r,4R)-4-hydroxycyclohexyl)carbamoyl)pyridin-3-yl )phenyl)-3-azabicyclo[3.1.0]hexane-3-carboxylate (intermediate 1b), except that 2-amino-5-(4-((1R,5S)-3-(4, 4-difluorocyclohexyl)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)nicotinic acid (intermediate 82a) instead of 2-amino-5-(4-((1R,5S)-3-( tert - butoxycarbonyl)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)nicotinic acid (intermediate 1c) and used trans -4-amino-1-methylcyclohexanol instead of trans -4-aminocyclohexanol. The crude product was purified by prepHPLC (Method 1a) to give the title compound, which was obtained as the TFA salt. ^1H NMR (400 MHz, DMSO–d6) δ 8.37 (s, 1H), 8.26 (d, 1H), 8.09 (d,1H), 7.57 (d, 2H), 7. 23 (d, 2H), 7.08 (s, 2H), 4.33 (s, 1H), 3.81 (bs, 1H), 3.39 (d, 2H), 2.62–2.56 (m, 2H), 2.00 (bs, 2H), 1.88–1.76 (m, 8H), 1.67–1.53 (m, 4H), 1.50–1.43 (m , 4H), 1.30 (t, 1H), 1.17 (s, 3H), 0.76 (bs, 1H). (UPLC–MS) t _R 0.61 min; ESI–MS 525 [M+H] ⁺ .

Intermediate 82a : 2-amino-5-(4-((1R,5S)-3-(4,4-difluorocyclohexyl)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl) nicotinic acid

The title compound was prepared in a manner similar to that for 2-amino-5-(4-((1R,5S)-3-( tert -butoxycarbonyl)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)nicotinic acid (intermediate 2c), except that methyl-2-amino-5-(4-((1R,5S)-3-(4,4-difluorocyclohexyl)-3-azabicyclo[3.1.0]hexane) was used –1-yl)phenyl)nicotinate (intermediate 82b) instead of (1R,5S)- tert -butyl-1-(4-(6-amino-5-(methoxycarbonyl)pyridin-3-yl)phenyl)-3- azabicyclo[3.1.0]hexane-3-carboxylate (intermediate 1d). (UPLC–MS) t _R 0.46 min; ESI–MS 414 [M+H] ⁺ .

Intermediate 82b : Methyl-2-amino-5-(4-((1R,5S)-3-(4,4-difluorocyclohexyl)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)nicotinate

The title compound was prepared in a manner similar to that for methyl-2-amino-5-(4-((1R,5S)-3-(tetrahydro-2H-pyran-4-yl)-3-azabicyclo[3.1.0]hexane –1-yl)phenyl)nicotinate (intermediate 14b), except that 4,4-difluorocyclohexanone was used instead of dihydro-2H-pyran-4(3H)-one. (UPLC–MS) t _R 0.72 min; ESI–MS 428 [M+H] ⁺ .

Example 83 : 2-amino-N-(3-(2-hydroxypropan-2-yl)bicyclo[1.1.1]pentan-1-yl)-5-(4-((1R,5S)-3-(tetrahydro –2H-pyran-4-yl)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)nicotinamide

The title compound was prepared in a manner similar to (1R,5S) -tert -butyl-1-(4-(6-amino-5-(((1r,4R)-4-hydroxycyclohexyl)carbamoyl)pyridin-3-yl )phenyl)-3-azabicyclo[3.1.0]hexane-3-carboxylate (intermediate 1b), except that 2-amino-5-(4-((1R,5S)-3-(tetrahydro- 2H-pyran-4-yl)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)nicotinic acid (intermediate 14a) instead of 2-amino-5-(4-((1R,5S)-3 –( tert -butoxycarbonyl)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)nicotinic acid (intermediate 1c) and 2-(3-aminobicyclo[1.1.1]pentan-1-yl)propane –2-ol hydrochloride (intermediate 83a) instead of trans –4-aminocyclohexanol. The crude product was purified by prepHPLC (Method 1a) to give the title compound. ^1H NMR (400 MHz, DMSO–d6) δ 8.98 (s, 1H), 8.37 (s, 1H), 8.15 (s, 1H), 7.57 (d, 2H), 7. 20 (d, 2H), 7.18 (s, 2H), 4.15 (s, 1H), 3.85–3.79 (m, 2H), 3.38 (bs, 1H), 3.08 (bs, 1H), 2.55–2.25 (m, 5H), 1.90 (s, 6H), 1.89–1.71 (m, 3H), 1.39–1.22 (m , 3H), 1.06 (s, 6H), 0.75 (bs, 1H). (UPLC–MS) t _R 0.59 min; ESI–MS 503 [M+H] ⁺ .

Intermediate 83a : 2-(3-aminobicyclo[1.1.1]pentan-1-yl)propan-2-ol hydrochloride

Stir tert -butyl(3-(2-hydroxypropan-2-yl)bicyclo[1.1.1]pentan-1-yl)carbamate (intermediate 83b, 298 mg, 1.235 mmol) in a saturated solution of HCl in ethanol (3.8 ml) for 29 hours at room temperature. The reaction mixture was concentrated under reduced pressure to give the title compound as the crude hydrochloride salt, which was used without further purification. ^1H NMR (400 MHz, DMSO–d6) δ 8.51 (s, 3H), 1.78 (s, 6H), 1.03 (s, 6H).

Intermediate 83b : tert -butyl(3-(2-hydroxypropan-2-yl)bicyclo[1.1.1]pentan-1-yl)carbamate

To a solution of methyl 3-(( tert -butoxycarbonyl)amino)bicyclo[1.1.1]pentane-1-carboxylate (679 mg, 2.81 mmol) in anhydrous THF, 3 M bromomethylmagnesium in THF (4.13 ml) was added dropwise , 12.4 mmol) at –78°C in an argon atmosphere. The reaction mixture was heated to room temperature. and it was stirred for 4 hours, then slowly diluted with 2 ml MeOH while adjusting the temperature. The mixture was concentrated under reduced pressure and the crude product was purified by normal phase chromatography (Method 2a) to give the title compound. (UPLC–MS) t _R 0.22 min; ESI–MS 142 [M+H] ⁺ indicated the loss of the BOC group under ionization conditions.

Example 84 : 2-amino-N-cyclohexyl-5-(3-(morpholine-4-carbonyl)phenyl)nicotinamide

The title compound was prepared in a manner similar to that for 2-amino-N-cyclohexyl-5-(4-(3-(piperidin-1-yl)propoxy)phenyl)nicotinamide (Example 43), except that 3- (morpholine-4-carbonyl)phenylboronic acid instead of 1-(3-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenoxy)propyl)piperidine. ^1H NMR (400 MHz, DMSO–d6) δ 8.45 (s, 1H), 8.38 (d, 1H), 8.18 (s, 1H), 7.78 (d, 1H), 7. 71 (s, 1H), 7.52 (t, 1H), 7.34 (d, 1H), 7.20 (s, 2H), 3.85–3.70 (m, 1H), 3.70 –3.50 (m, 8H), 1.90–1.80 (m, 2H), 1.80–1.70 (m, 2H), 1.70–1.55 (m, 1H), 1 .35–1.29 (m, 4H), 1.28–1.05 (m, 1H). (HPLC–MS) t _R 1.52 min; APCI–MS 409 [M+H] ⁺ .

Example 85 : 2-amino-5-(3-chloro-4-(((R)-2-methylpyrrolidin-1-yl)methyl)phenyl)-N-((1r,4R)-4-hydroxy-4- methylcyclohexyl)nicotinamide

The title compound was prepared in a manner similar to that for 2-amino-5-(3-fluoro-4-(((R)-2-methylpyrrolidin-1-yl)methyl)phenyl)-N-((1r,4R)- 4-hydroxy-4-methylcyclohexyl)nicotinamide (Example 46), except that (R)-2-(3-chloro-4-((2-methylpyrrolidin-1-yl)methyl)phenyl)-6- methyl-1,3,6,2-dioxazaborocane-4,8-dione (intermediate 85a) instead of (R)-2-(3-fluoro-4-((2-methylpyrrolidin-1-yl)methyl)phenyl) –6-methyl-1,3,6,2-dioxazaborocane-4,8-dione (intermediate 46a). (UPLC–MS) t _R 0.79 min; ESI–MS 457/459 [M+H] ⁺ .

Intermediate 85a : (R)-2-(3-chloro-4-((2-methylpyrrolidin-1-yl)methyl)phenyl)-6-methyl-1,3,6,2-dioxazaborocane-4,8- dione

The title compound was prepared in a manner similar to that for (R)-2-(3-fluoro-4-((2-methylpyrrolidin-1-yl)methyl)phenyl)-6-methyl-1,3,6,2-dioxazaborocane –4,8-dione (intermediate 46a), except that 2-chloro-4-(6-methyl-4,8-dioxo-1,3,6,2-dioxazaborocane-2-yl)benzaldehyde was used (intermediate 85b) instead of 2-fluoro-4-(6-methyl-4,8-dioxo-1,3,6,2-dioxazaborocan-2-yl)benzaldehyde (intermediate 46b). ESI–MS 365/367 [M+H] ⁺ .

Intermediate 85b : 2-chloro-4-(6-methyl-4,8-dioxo-1,3,6,2-dioxazaborocan-2-yl)benzaldehyde

The title compound was prepared in a manner similar to 2-fluoro-4-(6-methyl-4,8-dioxo-1,3,6,2-dioxazaborocan-2-yl)benzaldehyde (intermediate 46b), except that they used (3-chloro-4-formylphenyl)boronic acid instead of (3-fluoro-4-formylphenyl)boronic acid. ESI–MS 591 [2M+H] ⁺ .

Example 86 : 5-(4-((R)-1-(2-oxa-6-azaspiro[3.3]heptan-6-yl)ethyl)phenyl)-2-amino-N-((1r,4R)- 4-hydroxy-1-methylcyclohexyl)nicotinamide

To a solution of 2-amino-5-bromo-N-((1r,4r)-4-hydroxy-1-methylcyclohexyl)nicotinamide (intermediate 86a, 100 mg, 0.152 mmol) in dioxane (3 ml) was added (R)– 6-(1-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)ethyl)-2-oxa-6-azaspiro[3.3]heptane (intermediate 86b, 100 mg, 0.152 mmol), PdCl ₂ (dppf) (5.57 mg, 7.62 μmol) and 2 N. aq. NaOH (0.152 ml, 0.305 mmol) under nitrogen at room temperature. The reaction mixture was stirred at 80°C for 2 hours, then diluted with EtOAc and water. After two extraction steps with EtOAc, the combined organic layers were washed with sat. aq. NaHCO ₃ solution and saline solution, respectively. After drying over MgSO ₄ , filtration and concentration under reduced pressure, the crude product was purified by normal phase chromatography (method 2b) to give the title compound. ^1H NMR (400 MHz, DMSO–d6) δ 8.37 (s, 1H), 8.05 (bs, 1H), 7.70 (s, 1H), 7.62 (bs, 2H), 7. 38 (bs, 2H), 6.92 (bs, 2H), 4.62 (s, 4H), 4.47 (s, 1H), 3.48–3.42 (m, 1H), 3.40 –3.35 (m, 4H), 3.27–3.20 (m, 2H), 3.15 (bs, 1H), 1.60 (m, 2H), 1.65–1.16 (m , 13H). (UPLC–MS) t _R 0.52 min; ESI–MS 451 [M+H] ⁺ .

Intermediate 86a : 2-amino-5-bromo-N-((1r,4r)-4-hydroxy-1-methylcyclohexyl)nicotinamide

The title compound was prepared in a manner similar to 2-amino-5-bromo-N-((1r,4r)-4-hydroxycyclohexyl)nicotinamide (intermediate 3c), except that trans -4-amino-4 was used -methylcyclohexanol instead of trans -4-aminocyclohexanol. (UPLC–MS) t _R 0.70 min; ESI–MS 328/330 [M+H] ⁺ .

Intermediate 86b : (R)-6-(1-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)ethyl)-2-oxa-6- azaspiro[3.3]heptane

The title compound was prepared in a manner similar to 1-(3-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)-4-isopropylpiperazine (intermediate 8a), except that (R)-6-(1-(4-bromophenyl)ethyl)-2-oxa-6-azaspiro[3.3]heptane (intermediate 3b) was used instead of 1-(4-bromo- 3-fluorophenyl)-4-isopropylpiperazine (intermediate 8b). (UPLC–MS) t _R 0.70 min; ESI–MS 330 [M+H] ⁺ .

Example 87 : 2-amino-N-((1r,4R)-4-hydroxy-4-methylcyclohexyl)-5-(2-methyl-4-(((R)-2-methylpyrrolidin-1-yl)methyl) phenyl)nicotinamide

The title compound was prepared in a manner similar to that for 2-amino-5-(3-fluoro-4-(((R)-2-methylpyrrolidin-1-yl)methyl)phenyl)-N-((1r,4R)- 4-hydroxy-4-methylcyclohexyl)nicotinamide (Example 46), except that (R)-6-methyl-2-(2-methyl-4-((2-methylpyrrolidin-1-yl)methyl)phenyl was used )-1,3,6,2-dioxazaborocane-4,8-dione (intermediate 87a) instead of (R)-2-(3-fluoro-4-((2-methylpyrrolidin-1-yl)methyl)phenyl) –6-methyl-1,3,6,2-dioxazaborocane-4,8-dione (intermediate 46a). (UPLC–MS) t _R 0.65 min; ESI–MS 437 [M+H] ⁺ .

Intermediate 87a : (R)-6-methyl-2-(2-methyl-4-((2-methylpyrrolidin-1-yl)methyl)phenyl)-1,3,6,2-dioxazaborocane-4,8- dione

The title compound was prepared in a manner similar to that for (R)-2-(3-fluoro-4-((2-methylpyrrolidin-1-yl)methyl)phenyl)-6-methyl-1,3,6,2-dioxazaborocane –4,8-dione (intermediate 46a), except that 3-methyl-4-(6-methyl-4,8-dioxo-1,3,6,2-dioxazaborocan-2-yl)benzaldehyde was used (intermediate 87b) instead of 2-fluoro-4-(6-methyl-4,8-dioxo-1,3,6,2-dioxazaborocan-2-yl)benzaldehyde (intermediate 46b). ESI–MS 345 [M+H] ⁺ .

Intermediate 87b : 3-methyl-4-(6-methyl-4,8-dioxo-1,3,6,2-dioxazaborocan-2-yl)benzaldehyde

The title compound was prepared in a manner similar to 2-fluoro-4-(6-methyl-4,8-dioxo-1,3,6,2-dioxazaborocan-2-yl)benzaldehyde (intermediate 46b), except that they used (4-formyl-2-methylphenyl)boronic acid instead of (3-fluoro-4-formylphenyl)boronic acid. ESI–MS 568 [2M+H ₂ O+H] ⁺ .

Example 88 : 2-amino-N-cyclohexyl-5-(3-(4-(2-hydroxyethyl)piperazine-1-carbonyl)phenyl)nicotinamide

The title compound was prepared in a manner similar to that for 2-amino-N-cyclohexyl-5-(4-(3-(piperidin-1-yl)propoxy)phenyl)nicotinamide (Example 43), except that 3- (4-(2-hydroxyethyl)piperazine-1-carbonyl)phenylboronic acid instead of 1-(3-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenoxy) propyl)piperidine. ^1H NMR (400 MHz, DMSO–d6) δ 8.44 (s, 1H), 8.38 (d, 1H), 8.17 (s, 1H), 7.77 (d, 1H), 7. 68 (s, 1H), 7.51 (t, 1H), 7.31 (d, 1H), 7.19 (s, 2H), 4.44 (t, 1H), 3.85–3.70 (m, 1H), 3.70–3.58 (m, 2H), 3.58–3.48 (m, 2H), 3.48–3.35 (m, 2H), 2.50–2 .32 (m, 6H), 1.90–1.80 (m, 2H), 1.80–1.70 (m, 2H), 1.70–1.55 (m, 1H), 1.35 –1.29 (m, 4H), 1.28–1.05 (m, 1H). (HPLC–MS) t _R 1.31 min; APCI–MS 452 [M+H] ⁺ .

Example 89 : Salts, amorphous forms and crystalline polymorphs and pseudopolymorphs of the compound of Example 34.

A) Free form modification H _A. 800 g of the free form 2-amino-N-(4-hydroxybicyclo[2.2.2]octan-1-yl)-5-(4-((1R,5S)-3-(tetrahydro-2H-pyran-4-) were dissolved yl)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)nicotinamide in 3 liters of acetone. The resulting solution was stirred at 50°C and 2 liters of water were gradually added. The mixture was stirred at 50°C for 30 minutes, then 4 liters of water were gradually added. The mixture was stirred at 50°C for 2 hours and cooled to 22°C over a period of 2 hours. The solid was separated by suction filtration, washed twice with 1 liter of acetone:water 1:2 and dried at 45°C for 24 hours without applying a vacuum. We obtained approximately 660 g of the free form of the modification _HA 2-amino-N-(4-hydroxybicyclo[2.2.2]octan-1-yl)-5-(4-((1R,5S)-3-(tetrahydro-2H- pyran-4-yl)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)nicotinamide as a white solid. The free form of modification _HA is a monohydrate.

In fig. Figure 3 shows a free-form XRPD diagram of the modification _HA .

Table A below details the corresponding peaks and their 2-theta values ("angle"), as well as their relative intensity values (all 2-theta values are +/− 0.2).

Table A

Corner Rel. intensity 15.5 100.0% 17.5 87.6% 17.8 74.4% 7.4 68.7% 14.3 41.9% 18.6 35.8% 17.2 33.6% 12.4 32.7% 14.8 28.9% 16.8 27.7% 15.9 15.5% 9.4 14.4% 19.8 11.7% 24.0 11.7% 12.2 11.7% 20.7 10.9% 19.6 10.6% 24.7 10.5% 10.8 10.7%

In fig. Figure 4 shows an open differential scanning calorimetry (DSC) diagram of the free form modification _HA . The first endothermic peak in the DSC diagram corresponds to the dehydration of the free form of the _HA modification. The second endothermic peak corresponds to the melting of the anhydrous form resulting from dehydration.

In fig. Figure 5 shows a thermogravimetric analysis (TGA) diagram of the free form modification _HA .

The TGA results demonstrate that the free form of the H _A modification contains approximately 3.5% water by weight, which corresponds to approximately 1 molecule of water.

In fig. Figure 6 shows the FT–IR diagram of the free form modification _HA . Detected peaks:

wavenumber (in cm ⁻¹ ): 3481(w), 3328(m), 2931(m), 2886(w), 2863(w), 2787(w), 1632(s), 1617(m), 1584 (w), 1524(s), 1459(s), 1242(m), 1090(m), 885(m), 769(m). (s=strong, m=medium, w=weak).

B) Free form modification A. Added 0.5 g of the free form of modification _HA 2-amino-N-(4-hydroxybicyclo[2.2.2]octan-1-yl)-5-(4-((1R,5S)-3-(tetrahydro-2H –pyran-4-yl)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)nicotinamide in 3.5 ml of acetone. The mixture was stirred at 40°C for 1 hour and cooled to 22°C. The solid was filtered and dried at 40°C for 12 hours. Approximately 0.4 g of the free form of modification A was obtained as a white solid.

In fig. Figure 7 shows a free-form XRPD diagram of modification A.

Table B below details the corresponding peaks and their 2-theta values ("angle"), as well as their relative intensity values (all 2-theta values are +/− 0.2).

Table B

Rel. intensity 16.2 100.0% 17.8 85.3% 18.6 57.1% 20.5 50.2% 20.7 48.0% 20.3 47.4% 17.2 42.9% 16.9 41.6% 17.0 36.5% 12.4 31.9% 19.1 25.6% 19.6 21.0% 26.4 18.6% 24.7 16.6% 21.3 10.9% 22.4 10.6% 15.1 10.0% 25.5 9.8% 28.1 9.1% 10.2 8.9%

In fig. Figure 8 shows a free-form open differential scanning calorimetry (DSC) diagram of modification A.

In fig. 9 shows a thermogravimetric analysis (TGA) diagram of the free form of modification A. The TGA results indicate that the free form of modification A is an anhydride.

In fig. Figure 10 shows the FT-IR diagram of the free form of modification A. Detected peaks:

wavenumber (in cm ⁻¹ ): 3480(w), 3436(w), 3401(w), 3301(m), 2944(w), 2864(w), 2805(w), 1644(s), 1615 (m), 1583(w), 1520(s), 1459(m), 1247(m), 1098(m), 883(m), 801(m). (s=strong, m=medium, w=weak).

C) Free anhydride form. 100 mg of the free form of modification was subjected to H _A 2-amino-N-(4-hydroxybicyclo[2.2.2]octan-1-yl)-5-(4-((1R,5S)-3-(tetrahydro-2H-pyran –4-yl)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)nicotinamide exposed to an N ₂ atmosphere at 25°C for 6 hours. The resulting substance was characterized by XRPD under a protective atmosphere of N ₂ . The solid is the free form of the anhydride 2-amino-N-(4-hydroxybicyclo[2.2.2]octan-1-yl)-5-(4-((1R,5S)-3-(tetrahydro-2H-pyran- 4-yl)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)nicotinamide.

Figure 18 shows the XRPD diagram of this anhydride form.

D)Free trihydrate form. Subjected 100 mg of free form of modification H_A 2-amino-N-(4-hydroxybicyclo[2.2.2]octan-1-yl)-5-(4-((1R,5S)-3-(tetrahydro-2H-pyran-4-yl)-3- azabicyclo[3.1.0]hexan–1–yl)phenyl)nicotinamide exposed to 80% relative humidity at 25°C overnight. The solid was characterized by XRPD at 80% relative humidity at 25°C. The solid is the free form of 2-amino-N-(4-hydroxybicyclo[2.2.2]octan-1-yl)-5-(4-((1R,5S)-3-(tetrahydro-2H-pyran- 4-yl)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)nicotinamide.

Figure 19 shows the XRPD diagram of this trihydrate form.

E)Fumarate salt modification H _A .To 3.03 g of the free form 2-amino-N-(4-hydroxybicyclo[2.2.2]octan-1-yl)-5-(4-((1R,5S)-3-(tetrahydro-2H-pyran- 4-yl)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)nicotinamide was added to 30 ml of acetone:water (80:20, v/v). The mixture was stirred at 50°C for 0.5 hour and a clear solution was obtained. A solution of 708.4 mg of fumaric acid in 15 ml of acetone:water (80:20, v/v) was slowly added to the solution. The resulting suspension was gradually cooled to 25°C and stirring was continued for 12 hours. The solid was separated by suction filtration, washed once with 10 ml acetone and exposed to ambient conditions (approx. 25°C, RH 50%) overnight. We obtained 3.3 g of fumarate salt modification H_A 2-amino-N-(4-hydroxybicyclo[2.2.2]octan-1-yl)-5-(4-((1R,5S)-3-(tetrahydro-2H-pyran-4-yl)-3- azabicyclo[3.1.0]hexan–1–yl)phenyl)nicotinamide as a white solid. Fumarate salt modification H_A is a dihydrate.

In fig. 11 shows an XRPD diagram of the fumarate salt modification _HA .

Table C below details the corresponding peaks and their 2-theta values ("angle"), as well as their relative intensity values (all 2-theta values are +/− 0.2).

Table C

Corner Rel. intensity 15.6 100.0% 18.0 91.6% 14.2 89.4% 16.2 65.0% 7.0 65.1% 25.1 55.9% 19.2 56.0% 14.7 38.6% 18.2 37.3% 14.9 33.9% 22.2 31.4% 18.7 30.5% 19.4 29.4% 20.2 28.5% 21.0 28.3% 13.2 29.4% 25.5 26.1% 16.6 26.3% 20.8 25.7% 25.8 22.0% 5.2 10.9%

In fig. 12 shows a DSC diagram of the fumarate salt modification _HA . A broad endothermic peak up to 150°C corresponds to the dehydration of the fumarate salt of the _HA modification.

In fig. 13 shows a thermogravimetric analysis (TGA) diagram of the fumarate salt modification _HA . The TGA results show that the fumarate salt of the H _A modification contains approximately 5.9% water by weight, which corresponds to approximately 2 molecules of water.

In fig. Figure 14 shows the FT–IR diagram of the fumarate salt of the _HA modification. Detected peaks:

wavenumber (in cm ⁻¹ ): 3241(m), 2951(w), 2867(w), 1669(m), 1538(m), 1456(w), 1355(m), 1249(m), 1087 (w), 979(w), 885(w), 797(w). (s=strong, m=medium, w=weak).

In fig. 15 shows the DVS diagram of the fumarate salt modification _HA at 25°C. Based on this, it can be concluded that the fumarate salt of modification _HA is physically stable at an RH of at least 0% to 90% at 25°C.

In fig. 16 shows the DVS diagram of the fumarate salt modification _HA at 40°C. Based on this, it can be concluded that the fumarate salt of modification _HA is physically stable at an RH of at least 10% to 90% at 40°C.

It should be noted that the fumarate salt modification _HA is stable over a wider range of humidity than the free form polymorphs (data not shown).

F)Amorphous form of fumarate salt (option 1). Added 500 mg of fumarate salt modification H_A 2-amino-N-(4-hydroxybicyclo[2.2.2]octan-1-yl)-5-(4-((1R,5S)-3-(tetrahydro-2H-pyran-4-yl)-3- azabicyclo[3.1.0]hexan–1–yl)phenyl)nicotinamide in 10 ml of ethanol at 25°C to form a clear solution. Then 3 ml of heptane was added to the solution. A white solid precipitated. The solid was separated by suction filtration and dried at 40°C under vacuum overnight. We obtained 380 mg of the amorphous form of the fumarate salt 2-amino-N-(4-hydroxybicyclo[2.2.2]octan-1-yl)-5-(4-((1R,5S)-3-(tetrahydro-2H-pyran- 4-yl)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)nicotinamide as a white solid. The amorphous form exhibits a transition to a glassy state at approximately 143°C when analyzed using modulated DSC at a heating rate of 2 K/min, a temperature amplitude of 1 K, a period of 60 seconds.

G)Amorphous form of fumarate salt (option 2). The fumarate salt of modification H was heated_A 2-amino-N-(4-hydroxybicyclo[2.2.2]octan-1-yl)-5-(4-((1R,5S)-3-(tetrahydro-2H-pyran-4-yl)-3- azabicyclo[3.1.0]hexan–1–yl)phenyl)nicotinamide using DSC to 150°C at 10 K/min, then cooled to –20°C at 20 K/min. The resulting material was an amorphous form of the fumarate salt 2-amino-N-(4-hydroxybicyclo[2.2.2]octan-1-yl)-5-(4-((1R,5S)-3-(tetrahydro-2H-pyran –4-yl)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)nicotinamide, which was a white solid and exhibited a glassy state at ca. 78°C for DSC analysis at a heating rate of 10 K/min.

H)Phosphate salt.In 3.01 g of the free form 2-amino-N-(4-hydroxybicyclo[2.2.2]octan-1-yl)-5-(4-((1R,5S)-3-(tetrahydro-2H-pyran- 4-yl)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)nicotinamide was added to 30 ml of acetone:water (75:25, v/v). The resulting mixture was stirred at 50°C for 0.5 hour and a clear solution was obtained. A solution of 695.2 mg in 10 ml of acetone:water (95:5, v/v) was slowly added to the solution. The resulting suspension was gradually cooled to 25°C and stirring was continued for 24 hours. 1.5 ml of water was added to the suspension. Stirring of the mixture was continued at 50°C for 12 hours and it was slowly cooled to 25°C. The solid was separated by suction filtration, washed once with 10 ml acetone:water (90:10, v/v) and exposed to ambient conditions (approx. 25°C, RH 50%) overnight. We obtained 2.7 g of phosphate salt 2-amino-N-(4-hydroxybicyclo[2.2.2]octan-1-yl)-5-(4-((1R,5S)-3-(tetrahydro-2H-pyran- 4-yl)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)nicotinamide as a white solid.

In fig. Figure 17 shows the XRPD diagram of the phosphate salt.

Table D below details the corresponding peaks and their 2-theta values ("angle"), as well as their relative intensity values (all 2-theta values are +/− 0.2).

Table D

Corner Rel. intensity 21.7 100.0% 20.7 84.7% 16.2 78.4% 22.7 69.9% 11.9 69.5% 14.7 56.9% 19.2 55.2% 16.3 50.9% 20.4 48.0% 18.4 40.3% 19.5 36.5% 17.3 36.5% 12.4 35.3% 15.8 31.6% 18.5 27.9% 25.2 23.9% 24.3 21.7% 24.9 21.0% 23.6 20.1% 23.3 19.5% 27.6 15.4% 13.5 15.2% 6.8 15.0% 29.6 13.1% 7.4 9.8%

I) Fumarate salt modification H _A (F) above) shows higher solubility than the free form of modification H _A in some aqueous buffers.

Parameter Free form modification H _A Fumarate salt modification H _A Solubility (at 25°C, equilibrium 24 hours, target substance concentration = 2 mg/ml) Media/measured pH Solubility (mg/ml) pH Solubility (mg/ml) pH 1 n. HCl with pH 1.0/1.1 >2 1.03 >2 1.00 HCl based buffer with pH 2.0/2.1 >2 2.20 >2 2.09 Acetate buffer pH 4.7/4.8 >2 4.87 >2 4.68 Phosphate buffer pH 6.8/6.8 0.34 6.61 1.08 6.50 Borate buffer pH 9.0/8.6 <LOQ 8.45 0.03 8.05 Water/7.2 0.07 9.04 >2 4.57 SGF with pH 2.0/2.0 >2 2.87 >2 2.43 FaSSIF pH 6.5/6.5 0.47 6.47 1.69 6.09 FeSSIF pH 5.8/5.9 1.69 5.92 >2 5.67

(LOQ=limit of quantitation;

FaSSIF=Simulated intestinal juice on an empty stomach;

FeSSIF=Simulation of intestinal juice after eating;

SGF=Simulated gastric juice.

FeSSIF

Excipients Quantity per 100 ml Sodium taurocholate [mg] 538.7 Water [mg] 3500 Lipoid E PCS [mg] 155 Glyceryl monooleate
Rylo MG 19 PHARMA [mg] 178.3 Oleate Na [mg] 24.4

FaSSIF

Excipients Quantity per 100 ml Sodium taurocholate [mg] 161.4 Water [mg] 1000 Lipoid E PCS [mg] 15.5 Buffer FaSSIF Add until final volume is reached

Retrieving the FaSSIF Buffer

Excipients Quantity per 100 ml NaCl [g] 0.401 Maleic acid [g] 0.222

SGF

Excipient Quantity/volume per 1 liter NaCl 2 g Triton X-100 1 g 0.1 M HCl 100 ml

J) Lactic acid salt, tartaric acid salt and malic acid salt of Compound A can be prepared by methods similar to those above.

K) Various solvates can be prepared from many of the salts and salt forms of Compound A above (not shown in detail).

The XRPD, TGA, DSC, FT–IR and DVS data mentioned in the examples above were obtained using the following methods.

a) TGA method

Device: TA Discovery, TA Discovery, New Castle, Delaware, USA

Temperature range: 30 to 300°C

Scanning speed: 10°C/min.

Nitrogen flow: 20 ml/min.

Sample weight: ~2–10 mg

b) DSC method

Device: TA Discovery

Temperature range: 30 to 250 or 300°C

Scanning speed: 10°C/min.

Nitrogen flow: 50 ml/min.

Sample weight: ~2 mg

c) XRPD method. All XRPD diagrams were obtained in reflected light mode.

(i) XRPD Method 1 (Fig. 3, 11, 17)

Device: Bruker D8 Advance (Bruker, Bruchsal, Germany))

Detector: LYNXEYE (1D mode), open angle: 1.996°

Emission: CuKα (0.15406 nm)

Monochromator: nickel filter

X-ray generator power: 40 kV, 40 mA

Step size, separation: 0.041 degrees

Scan range: 2° to 45° (2-theta value)

Scan duration: 36 min.

Slits: primary: fixed illuminated sample size: 5 mm, secondary slit: 5 mm, Soller corner slit: 2.5°

(ii) XRPD Method 2 (Fig. 7)

Device: Bruker D8 Advance

Detector: LYNXEYE (1D mode), open angle: 1.996°

Emission: CuKα (0.15406 nm)

Monochromator: nickel filter

X-ray generator power: 40 kV, 40 mA

Step size, separation: 0.041 degrees

Scan range: 2° to 45° (2-theta value)

Scan duration: 330 seconds

Slits: Primary: Fixed illuminated sample size: 5mm, Secondary slit: 5mm, Soller corner slit: 2.5

XRPD method 3 (Fig. 18, 19)

Device: Bruker D8 Advance XRPD with Cryo–RH chamber (Bruker, Bruchsal, Germany)

Detector: VANTEC–1 (1D detector)

Emission: CuKα (0.15406 nm)

Monochromator: nickel filter

X-ray generator power: 40 kV, 40 mA

Step size, separation: 0.0165 degrees

Scan range: 2° to 45° (2-theta value)

Scan duration: 17 min.

Slits: scattering slit: V12, anti-scattering slit: 10.0 mm, detector slit: 17.43 mm,

primary Soller gap: 2.5°, secondary Soller gap: 2.5°

d) DVS

Device: Advantage (Surface Measurement Systems, London, UK)

Sample weight: ~10 mg

Temperature: 25°C

dm/dt: 0.002%/min.

e) FT-IR

Thermo Fisher Nicolet 6700 Attenuated Total Reflectance (ATR), Thermo Fisher Scientific, Waltham, MA, USA.

Biochemical tests (examples 90 and 91)

For all biochemical analyses, recombinant human proteins were expressed in and purified from baculovirus-transfected insect cells. The constructs contained the GS domain and kinase domain of wild-type ALK2 (aa172–499), FOP mutant ALK2 (aa172–499 R206H), ALK3 (aa198–525), ALK5 (aa162–503), and ALK6 (aa168–495).

Example 90: In Vitro Enzyme Inhibition Assay Using a Biochemical Autophosphorylation Assay (ADPGlo Luminescence-Based Kinase Activity Assay) - "ADPGlo Assay"

The kinase selectivity panel, which measures autophosphorylation using the ADP-Glo™ Kinase Activity Assay Kit (Promega, V9101), was set up for wild-type ALK2 (aa172–499) and ALK3 (aa198–525).

Analyzes were carried out in 384-well small-volume analytical microtiter plates with a final volume of the reaction mixture of 6 μl. Dose-response curves were obtained by incubating 10 nM of each kinase in 50 mM Hepes pH 7.5, 0.02% Tween 20, 0.02% BSA, 1 mM DTT, 10 µM Na ₃ VO ₄ , 10 mM β-glycerol phosphate , 1 mM MgCl ₂ , 12 mM MnCl ₂ and 15 µM ATP for 60 min. at 32°C in the presence or absence of compound diluted in DMSO. The amount of ADP produced is a measure of kinase activity and was quantified using the ADP-Glo™ Kinase Activity Assay Kit (Promega) according to the manufacturer's instructions. ADP is converted to ATP by adding 3 µl of ADP–Glo™ reagent and incubating at 32°C for 60 min. ATP is subsequently converted to a bioluminescent signal by the addition of 6 μl of luciferase test kit reagents (kinase detection buffer+kinase detection substrate (Promega)) and further incubation at 32°C for 60 min. To measure luminescence, a PHERAstar™ multi-channel reader was used with a measurement interval time of 0.1 seconds (optical module for luminescence measurements in the wavelength range from 230 nm to 750 nm). The luminescent signal was positively correlated with kinase activity.

The specific activity values are shown in the table below.

Note IC ₅₀ ALK2 with ADPGlo [µmol l ^-1 ] IC ₅₀ ALK3 with ADPGlo [µmol l ^-1 ] 1 0.005 0.310 2 0.012 1,767 3 0.012 1,050 4 0.005 0.830 5 0.007 1,100 6 0.018 0.940 7 0.012 0.875 8 0.013 1,900 9 0.011 1,600 10 0.011 1,400 eleven 0.009 1,100 12 0.014 3,100 13 0.008 1,400 14 0.018 4,100 15 0.029 0.967 16 0.012 1,400 17 0.012 2,000 18 0.014 2,300 19 0.012 1,800 20 0.007 1,350 21 0.006 0.260 22 0.009 1,500 23 0.009 1,600 24 0.011 6,475 25 0.009 1,300 26 0.018 1,700 27 0.010 1,300 28 0.011 0.530 29 0.022 2,500 thirty 0.007 0.500 31 0.013 1,200 32 0.022 12,700 33 0.010 2,400 34 0.018 0.914 35 0.017 0.903 36 0.016 1,450 37 0.008 0.960 38 0.007 1,300 39 0.013 0.830 40 0.032 0.995 41 0.019 1,800 42 0.026 1,300 43 0.031 2,350 44 0.018 0.430 45 0.020 1,600 46 0.024 14,600 47 0.021 1,300 48 0.008 0.970 49 0.027 1,600 50 0.011 2,400 51 0.016 0.780 52 0.067 2,800 53 0.013 1,200 54 0.011 0.920 55 0.022 2,440 56 0.028 9,900 57 0.038 1.185 58 0.025 3.725 59 0.024 2,700 60 0.011 0.990 61 0.043 2,100 62 0.024 4,500 63 0.091 1,800 64 0.110 2,200 65 0.082 2,400 66 0.015 1,200 67 0.022 1,900 68 0.034 8,000 69 0.041 3,100 70 0.020 2,850 71 0.022 2,900 72 0.046 9,900 73 0.075 1,900 74 0.064 13,200 75 0.011 1,800 76 0.034 3,200 77 0.028 3,300 78 0.037 8,300 79 0.086 6,500 80 0.058 1,800 81 0.150 15,100 82 0.082 9,500 83 0.100 15,100 84 2,400 21,900 85 0.190 15,100 86 0.110 8,200 87 0.880 15,100 88 0.990 22,800

Example 91. In vitro enzyme inhibition assay using a biochemical peptide phosphorylation assay - "Caliper analysis"

The kinase selectivity panel, which measures the level of phosphorylation of a peptide substrate, was set up for wild-type ALK2 (aa172-499), FOP mutants ALK2 (aa172-499 R206H), ALK1 (aa166-493), ALK5 (aa162–503) and ALK6 (aa168–495). The technology used for the described analysis is based on the separation and quantification of substrate and product in an electric field. During the reaction carried out by the kinase, the substrate, which is a peptide, is phosphorylated by the kinase. The transfer of the phosphate residue also introduces two additional negative charges and therefore changes the overall charge of the phosphopeptide compared to the non-phosphorylated peptide. Due to this charge difference, phosphorylated and unphosphorylated peptides migrate in the electric field at different speeds.

In the method used, this separation occurs inside a chip, which contains a complex capillary system for simultaneous analysis of 12 samples (“12-channel chip”, Caliper Technologies Corp., Mountain View, USA). To enable detection and quantification of peptides in the capillary system, the peptides were labeled with a fluorescent label (fluorescein). With this label present, the peptides can be quantified from fluorescence intensity using laser and detection system instruments (LC3000, Caliper Life Sciences).

Analyzes were carried out in 384-well small-volume analytical microtiter plates with a final volume of the reaction mixture of 9 μl. Dose-response curves were obtained by incubating 10 nM of each kinase together with 2 μmol of fluorescently labeled substrate, peptide 5-Fluo-Ahx-KKYQAEEN-T-YDEYENKK-amide (10 mM stock solution in DMSO), in 50 mM Hepes with pH 7.5, 0.02% Tween 20, 0.02% BSA, 1 mM DTT, 10 μM Na ₃ VO ₄ , 10 mM β-glycerol phosphate, 1 mM MgCl ₂ , 12 mM MnCl ₂ (for 7 mM ALK1 and ALK6) and 15 µM ATP for 60 min at 30°C in the presence or absence of the compound diluted in DMSO.

The kinase reaction was stopped by adding 15 μl of STOP buffer (100 mM HEPES pH 7.5, 5% DMSO, 0.1% Caliper coating reagent, 10 mM EDTA, and 0.015% Brij35).

Plates in which kinase reactions were stopped were moved to a Caliper LC3000 workstation (Caliper Technologies Corp., Mountain View, USA) for reading. The relative amount of phosphorylated peptide r was calculated using the values of the substrate peak height, s, and the product peak height, p: r=p/(p+s).

The specific activity values are shown in the table below.

Example EPK ALK2 (IC ₅₀ [µmol.
l ^-1 ]) EPK ALK2 FOP (IC ₅₀ [µmol.
l ^-1 ]) EPK ALK1 (IC ₅₀ [µmol.
l ^-1 ]) EPK ALK5 (IC ₅₀ [µmol.
l ^-1 ]) EPK ALK6 (IC ₅₀ [µmol.
l ^-1 ]) 1 0.009 0.008 0.075 0.560 0.740 2 0.011 0.011 0.093 2,167 2,500 3 0.011 0.015 0.136 2.133 1,733 4 0.013 0.007 0.175 2,450 1,850 5 0.010 0.011 0.081 2,200 2,500 6 0.015 0.028 0.220 10,000 5,200 7 0.012 0.008 0.195 5,450 2,400 8 0.012 0.022 0.220 4,000 5,000 9 0.011 0.014 0.127 3.133 3.133 10 0.011 0.013 0.089 2,600 2,500 eleven 0.012 0.012 0.150 2,400 3,900 12 0.012 0.021 0.160 1,900 2,400 13 0.013 0.016 0.140 3,100 3,000 14 0.016 0.030 0.710 10,000 10,000 15 0.020 0.029 0.370 3,767 2.233 16 0.015 0.018 0.240 9,900 5,000 17 0.014 0.014 0.110 1,600 2,400 18 0.015 0.025 0.200 2,700 2,100 19 0.018 0.014 0.105 1,850 2,250 20 0.017 0.011 0.130 3,800 3,950 21 0.019 0.010 0.055 0.900 1,000 22 0.016 0.014 0.250 2,500 3,200 23 0.018 0.019 0.250 2,700 2,400 24 0.016 0.021 0.543 6,800 7,300 25 0.018 0.011 0.130 1,400 3,000 26 0.018 0.024 0.230 4,300 2,500 27 0.018 0.014 0.135 1,300 1,600 28 0.018 0.017 0.250 9,400 3,300 29 0.020 0.039 0.280 4,000 4,000 thirty 0.022 0.014 0.210 4,000 2,800 31 0.020 0.017 0.230 3,500 4,500 32 0.020 0.027 0.743 3,200 3,300 33 0.020 0.014 0.220 1,800 2,000 34 0.023 0.021 0.323 6,280 3,300 35 0.021 0.018 0.283 5.725 4,500 36 0.023 0.024 0.295 10,000 7,200 37 0.027 0.008 0.100 1,100 1,700 38 0.028 0.007 0.160 1,500 2,600 39 0.024 0.019 0.190 1,600 2,000 40 0.029 0.049 0.410 3,900 2,850 41 0.023 0.027 0.310 6,700 6,900 42 0.029 0.036 0.540 4,767 4,100 43 0.025 0.026 0.765 3,500 11,050 44 0.024 0.025 0.170 3,200 2,000 45 0.025 0.027 0.750 2,700 4,800 46 0.025 0.029 1,200 10,000 10,000 47 0.025 0.027 0.290 4,700 3,500 48 0.034 0.011 0.120 5,300 4,400 49 0.027 0.033 0.590 3,400 8,800 50 0.037 0.014 0.500 6,400 5,100 51 0.031 0.018 0.120 1,400 1,700 52 0.031 0.056 0.970 8,600 20,200 53 0.032 0.020 0.190 2,000 2,500 54 0.033 0.018 0.140 5,300 3,500 55 0.030 0.033 0.627 5,033 2,900 56 0.030 0.038 0.930 5,800 10,000 57 0.035 0.051 0.485 4,400 3,200 58 0.032 0.032 0.663 7,550 8,400 59 0.033 0.038 0.770 9,100 7,700 60 0.042 0.019 0.170 6,300 6,300 61 0.035 0.046 0.940 7,200 20,200 62 0.036 0.043 0.840 8,800 10,000 63 0.057 0.140 0.940 6,700 3,500 64 0.063 0.160 1,000 7,500 3,800 65 0.044 0.034 0.580 1,700 10,000 66 0.056 0.026 0.190 10,000 6,200 67 0.050 0.034 0.180 10,000 8,700 68 0.049 0.067 2,500 10,000 10,000 69 0.052 0.042 1,100 5,200 9,000 70 0.068 0.037 0.305 9,400 8,750 71 0.065 0.032 0.490 3,000 5,800 72 0.061 0.068 2,950 10,000 10,000 73 0.061 0.091 0.640 6,900 4,000 74 0.070 0.092 2,500 10,000 10,000 75 0.140 0.018 0.210 4,600 7,700 76 0.079 0.055 0.370 9,400 8,400 77 0.080 0.059 0.320 10,000 8,600 78 0.109 0.043 0.310 7,800 10,000 79 0.087 0.053 0.890 4,100 10,000 80 0.087 0.087 0.680 6,600 4,600 81 0.110 0.150 3,000 10,000 10,000 82 0.117 0.091 1,600 10,000 10,000 83 0.145 0.180 5,600 10,000 10,000 84 0.350 1,800 20,200 20,200 20,200 85 0.170 0.210 9,800 10,000 10,000 86 0.205 0.190 2,300 10,000 10,000 87 0.630 0.900 10,000 10,000 10,000 88 0.810 0.950 20,200 20,200 20,200

Example 92 BMP Signal Transduction Assay Using a Reporter Gene

A human hepatocellular carcinoma cell line (HuH7) stably transfected with a reporter plasmid consisting of a BMP response element (BRE) from the Id1 promoter fused to a luciferase reporter gene was generated by lentiviral transduction.

Cells were maintained in DMEM ( GIBCO no. 41965 high glucose plus L-glutamine), 10% FCS (Amimed no. 2-01F10-I), 1% Pen/Strp (Amimed no. 4-01F00), and 5 μg/ml blastidicin ( InvivoGen No. ant–bl–1) at 37°C, 5% CO ₂ . Assays were performed in 384-well flat-bottomed polystyrene microtiter plates (cell culture-treated) with a sterile lid. 16 h before the start of the assay, cells were starved by replacing the medium with medium without blasticidin and FCS. Before analysis, cells were separated from the stock culture flask using trypsin/EDTA and counted. Cell suspension was obtained in the same medium without blasticidin and FCS. 2 x 10 ⁴ cells were added to each well of a plate already containing serial dilutions of each compound in DMSO (final DMSO concentration 0.5%) in a total volume of 40 μl. Cells and compound were incubated for 1 hour at 37°C, 5% CO ₂ , followed by stimulation with recombinant BMP6 (R&D Systems #507-BP/CF) at 5 μl per well at a final concentration of 100 ng/ml. The assay plates were incubated for an additional 5 hours at 37°C, 5% CO ₂ , after which luciferase levels were measured.

Quantification of expressed luciferase was performed using the Steady–Glo® Luciferase Assay System (Promega #E2520). 5 µl of Steady-Glo® reagent was added to each well, the samples were mixed by vigorously shaking the plate, after which luminescence was measured on a PHERAstar™ multichannel reader in 1 second/well mode (optical module for luminescence measurements in the wavelength range from 230 nm to 750 nm).

The specific activity values of the compounds of the present invention are further described in the table below.

Note IC ₅₀ MSD BMP RGA
[µmol l ^-1 ] 1 0.076 2 0.021 3 0.062 4 0.108 5 0.120 6 0.870 7 0.051 8 0.069 9 0.043 10 0.061 eleven 0.051 12 0.125 13 0.078 14 0.205 15 0.074 16 0.079 17 0.150 18 2,000 19 0.021 20 0.040 21 0.035 22 0.041 23 0.023 24 0.081 25 0.035 26 0.110 27 0.059 28 0.017 29 0.685 thirty 0.068 31 0.115 32 0.093 33 0.026 34 0.049 35 0.073 36 0.043 37 0.037 38 0.058 39 0.078 40 0.075 41 0.158 42 0.235 43 0.130 44 0.365 45 0.365 46 0.120 47 0.130 48 0.360 49 0.790 50 0.825 51 0.064 52 0.340 53 0.050 54 0.395 55 0.543 56 0.285 57 0.144 58 0.143 59 0.405 60 0.084 61 2,300 62 0.134 63 0.390 64 0.380 65 0.415 66 0.910 67 1.165 68 0.950 69 1.105 70 0.513 71 0.086 72 0.190 73 0.130 74 0.550 75 0.330 76 2,300 77 1,300 78 0.220 79 2,700 80 0.058 81 1,250 82 0.056 83 0.360 84 9,500 85 0.860 86 1,080 87 3,550 88 9,300

Example 93 In Vivo Efficacy in a Mouse Model of FOP (Fibrodysplasia Ossificans Progressiva) in Children

Universal heterozygous expression of the BMP type I receptor mutant Alk2 R206H, which mediates FOP, in mice leads to perinatal mortality (Chakkalakal et al., 2012).

To study the role of Alk2(R206H) in the postnatal period in the body of a young individual, a mutant mouse with inducible Alk2(R206H) (Prof. D. Goldhamer, UConn, ASBMR abstract, 2013) with an integrated stop cassette flanked is required as a preclinical mouse model of FOP. loxP sites located upstream of the mutant exon, which makes the expression of the mutant allele dependent on Cre recombinase. They were crossed with Rosa26–CreERt2 mice, which allowed for temporal and spatial regulation of heterozygous Alk2(R206H) expression.

In this case, mice heterozygous for the universally inducible Alk2(R206H) were used in combination with tamoxifen-inducible CreER–loxP technology (tam). Successful induction of Alk2(R206H) expression was achieved by administering tamoxifen to 1-week-old male and female Alk2(R206H) mice; Rosa26–CreERt2 .

FOP flares were induced by deep muscle trauma, achieved by local intramuscular (im.) injection of cardiotoxin (CTX) and concomitant intramuscular adenovirus injection, producing Alk2(R206H) -dependent local FOP flares. CTX causes skeletal muscle damage by inhibiting Ca/Mg–ATPase in the plasma membrane and inducing calcium release from the sarcoplasmic reticulum.

Body weight of 1-week-old Alk2(R206H) males and females; Rosa26–CreERt2 was measured at the beginning of the experiment and then daily. All pups were treated once daily with a subcutaneous (sc.) injection of 20 mg/kg tamoxifen (tamoxifen free base (Sigma T5648) dissolved in 10% absolute alcohol, 90% sunflower oil (Sigma S5007), at 8 mg/kg ml), starting from the age of 1 week for 10 days.

On day 3 of tamoxifen injection, all pups were subjected to local muscle injury by a single intramuscular injection of 100 μl adenovirus+cardiotoxin (CTX) into the gastrocnemius muscle of the right hind limb, taking care not to inject into blood vessels or touch the needle to bone. For this purpose, 90 μl of adenovirus (Ad–GFP–2A–iCre, cat. no. 1772, Vector Biolabs; titer: 1×10 ¹⁰ PFU (plaque forming units)/ml) was mixed with 10 μl of 100 μM cardiotoxin stock solution (derived from cardiotoxin , Cat# L8102, Latoxan, or Cardiotoxin, Cat# C9759, Sigma, using sterile saline). Animals were anesthetized using low-dose inhalation of isoflurane, the right hind limb was shaved and the skin was disinfected using betaseptic, followed by intramuscular injection, which was administered slowly and carefully using an insulin syringe.

Therapeutic oral treatment of mice heterozygous for Alk2(R206H) at bid frequency was started 3 days after muscle injury and was carried out with 2, 5, 10 and 25 mg/kg of compound A (compound from example 34=2-amino-N- (4-hydroxybicyclo[2.2.2]octan-1-yl)-5-(4-((1R,5S)-3-(tetrahydro-2H-pyran-4-yl)-3-azabicyclo[3.1.0] hexane-1-yl)phenyl)nicotinamide) or an appropriate carrier medium for 6 weeks. In all mice, heterotopic ossification was assessed biweekly using whole body and hindlimb radiography (Faxitron device) and in vivo imaging using microcomputed tomography (μCT) starting 2 weeks after Ad/CTX application.

To determine the full extent of HO, right hindlimbs were collected at necropsy for ex vivo micro-CT imaging. The entire right hind limb with attached muscle was cut as quickly as possible and transferred to 70% ethanol at 4°C. After 24 hours, samples were transferred to fresh 70% ethanol for high-resolution μCT imaging using the Scanco Medical μCT40 (voxel size: 16 μm, integration time: 250 ms, average resolution, 55 E(kVp), 145 μA ,mode: continuous cone beam rotation, threshold 275, Gaussian filter: standard deviation 0.7, neighborhood 1). The total volume of bone tissue (mm ³ ) of ectopic bone in the hind limb was determined. The results are shown in Figure 1. From the figure it can be seen that the use of compound A (compound from example 34) led to a decrease in bone volume of the right hind limb ex vivo .

To examine the effect of compound A (compound from Example 34) on the progression of heterotopic ossification after initiation of the process, mice heterozygous for Alk2(R206H) were similarly injected with 100 μl of adenovirus + cardiotoxin (CTX), but allowed heterotopic bone to develop for 2.5 weeks. After confirmation of HO production after 2 weeks, mice were randomized based on the presence of HO, genetic background, and sex. Treatment began 2.5 weeks post-injury with vehicle or 10 mg/kg Compound A bid and continued for 6 weeks. In all mice, heterotopic ossification was assessed biweekly using whole body and hindlimb radiography (Faxitron device) and in vivo imaging using microcomputed tomography (μCT) starting 2 weeks after Ad/CTX application.

To determine the full extent of HO, right hindlimbs were collected at necropsy for ex vivo micro-CT imaging. The entire right hind limb with attached muscle was cut as quickly as possible and transferred to 70% ethanol at 4°C. After 24 hours, samples were transferred to fresh 70% ethanol for high-resolution μCT imaging using the Scanco Medical μCT40 (voxel size: 16 μm, integration time: 250 ms, average resolution, 55 E(kVp), 145 μA ,mode: continuous cone beam rotation, threshold 275, Gaussian filter: standard deviation 0.7, neighborhood 1). The total volume of bone tissue (mm ³ ) of ectopic bone in the hind limb was determined. The results are shown in Figure 2. The results show that the use of Compound A (Compound of Example 34) prevented the increase in bone volume of the right hind limb ex vivo.

From the results shown in Figures 1 and 2, it is apparent that the compound of the present invention (Compound A) inhibited heterotopic ossification when dosed shortly after muscle injury and prevented further progression of heterotopic ossification in litters of ALK2(R206H) mice when administered doses were administered after heterotopic ossification had already begun.

Example 94. Heterotopic ossification in rats caused by tenotomy of the Achilles tendon at the midpoint

To test whether the compounds of the present invention are capable of preventing trauma-induced heterotopic ossification (HO) of soft tissue, therapeutic efficacy can be applied in a rat model of unilateral midpoint Achilles tendon tenotomy (Rooney et al. , Matrix 12: 274–281, 1992) . For this purpose, the left Achilles tendon of 8-week-old female Wistar rats (body weight range 190–265 g) was completely dissected using a sterile scalpel (blade number 11) under inhalational anesthesia with isoflurane, accompanied by analgesic treatment with 0.03 mg. /kg buprenorphine for 48 hours every 10–12 hours subcutaneously. Prophylactic oral treatment with a compound of the present invention (10 mg/kg qd) or vehicle (sodium carboxymethylcellulose:water:Tween 80, 0.5:99:0.5) was administered for 10 weeks starting from the day of surgery (n=11–12 rats per group). After surgery, rats were housed individually for 3–4 days and then housed in groups of two animals per cage at 25°C on a 12:12 h light/dark cycle and fed a standard rodent diet containing 18.2% protein and 3.0% fat, with a calorific value of 15.8 MJ/kg (3890, Provimi Kliba SA), with food and water provided ad libitum . The effectiveness of the treatment was assessed longitudinally by obtaining radiographs of the operated distal limb (Faxitron LX-60 system) at 4 and 10 weeks after tenotomy. Heterotopic bone volume was quantified in vivo using microcomputed tomography (micro-CT) under isoflurane inhalation anesthesia (vivaCT40 device, Scanco Medical AG; resolution 17.5 μm) at 6 and 9 weeks after surgery.

Claims (22)

1. A compound representing the hydrate of 2-amino-N-(4-hydroxy-bicyclo[2.2.2]octan-1-yl)-5-(4-((1R,5S)-3-(tetrahydro-2H- pyran-4-yl)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)nicotinamide or a pharmaceutically acceptable salt thereof. 2. The compound according to claim 1, where the compound is in free form. 3. A compound according to claim 1 or 2, wherein the compound is a monohydrate. 4. The compound of claim 3, wherein the compound is a free form modification of H _A characterized by an XRPD (X-ray powder diffraction) pattern shown in Figure 3. 5. A compound which is a pharmaceutically acceptable salt of 2-amino-N-(4-hydroxy-bicyclo[2.2.2]octan-1-yl)-5-(4-((1R,5S)-3-(tetrahydro- 2H-pyran-4-yl)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)nicotinamide, wherein the pharmaceutically acceptable salt is selected from the group consisting of fumarate, phosphate, lactate, tartrate and malic acid salt. 6. The compound according to claim 5, which is 2-amino-N-(4-hydroxy-bicyclo[2.2.2]octan-1-yl)-5-(4-((1R,5S)-3-(tetrahydro -2H-pyran-4-yl)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)nicotinamide phosphate, characterized by the XRPD (X-ray powder diffraction) pattern shown in Figure 17. 7. The compound according to claim 1, which is a trihydrate. 8. The compound according to claim 7, characterized by an XRPD (X-ray powder diffraction) pattern shown in Figure 19. 9. The compound is the anhydrate of 2-amino-N-(4-hydroxy-bicyclo[2.2.2]octan-1-yl)-5-(4-((1R,5S)-3-(tetrahydro-2H- pyran-4-yl)-3-azabicyclo[3.1.0]hexan-1-yl)phenyl)nicotinamide or a pharmaceutically acceptable salt thereof. 10. The compound of claim 9, wherein the compound is free form modification A characterized by the XRPD (X-ray powder diffraction) pattern shown in Figure 7. 11. The compound of claim 9, wherein the compound is characterized by an XRPD (X-ray powder diffraction) pattern shown in Figure 18. 12. Pharmaceutical composition for suppressing the activity of the receptor activin receptor-like kinase 2 (ALK-2) in a subject, containing a therapeutically effective amount of a compound according to any one of claims 1 to 11 and one or more pharmaceutically acceptable carriers. 13. Method of inhibiting the activity of the receptor activin receptor-like kinase 2 (ALK-2) in a subject, comprising administering to the subject a therapeutically effective amount of a compound according to any one of claims 1 to 11 or a pharmaceutical composition according to claim 12. 14. A method of treating a disorder or disease selected from heterotopic ossification or fibrodysplasia ossificans progressiva, comprising administering to a subject a therapeutically effective amount of a compound of any one of claims 1 to 11 or a pharmaceutical composition of claim 12. 15. The method of claim 14, wherein the disorder is heterotopic ossification. 16. The method of claim 14, wherein the disorder is fibrodysplasia ossificans progressive. 17. A compound according to any one of claims 1 to 11 for use as a medicament for the treatment of a disorder or disease selected from heterotopic ossification or fibrodysplasia ossificans progressiva. 18. A compound for use according to claim 17, wherein the disorder or disease is heterotopic ossification. 19. A compound for use according to claim 17, wherein the disorder or disease is fibrodysplasia ossificans. 20. Use of a compound according to any one of claims 1 to 11 for the preparation of a medicament for the treatment of a disorder or disease selected from heterotopic ossification or fibrodysplasia ossificans progressiva. 21. Use according to claim 20, wherein the disorder or disease is heterotopic ossification. 22. Use according to claim 20, wherein the disorder or disease is fibrodysplasia ossificans.