HK1178534A

HK1178534A - Pyrrolopyrazine derivatives and their use as jak and syk inhibitors

Info

Publication number: HK1178534A
Application number: HK13106246.4A
Authority: HK
Inventors: Robert Than Hendricks; Johannes Cornelius Hermann; Rama K. Kondru; Yan Lou; Stephen M. Lynch; Timothy D. Owens; Michael Soth
Original assignee: F. Hoffmann-La Roche Ag
Priority date: 2010-03-22
Filing date: 2011-03-21
Publication date: 2013-09-13

Description

Pyrrolopyrazine derivatives and their use as JAK and SYK inhibitors

The present invention relates to the use of novel pyrrolopyrazine derivatives which are JAK and SYK inhibitors, and which selectively inhibit JAK3 and are useful in the treatment of autoimmune and inflammatory diseases.

Protein kinases constitute one of the largest families of human enzymes and regulate many different signaling processes by adding phosphate groups to proteins; in particular tyrosine kinases phosphorylate proteins on the alcohol moiety of tyrosine residues. The tyrosine kinase family includes members that control cell growth, migration, and differentiation. Abnormal kinase activity has been implicated in a variety of human diseases including cancer, autoimmune and inflammatory diseases. Since protein kinases belong to key regulators of cell signaling, they provide a means to modulate cellular function with small molecule inhibitors of kinase activity and thus are good targets for drug design. In addition to treating kinase-mediated disease processes, selective and potent inhibitors of kinase activity may also be used to study cell signaling processes and identify other cellular targets of therapeutic interest.

JAKs (JAnus kinases) are a family of cytoplasmic protein tyrosine kinases including JAK1, JAK2, JAK3 and TYK 2. Each JAK associates preferentially with the intracytoplasmic portion of a discrete cytokine receptor (annu. rev. immunol.16(1998), pp.293-322). Following ligand binding, JAKs are activated and signal transduction is initiated by phosphorylating cytokine receptors, which themselves lack intrinsic kinase activity. This phosphorylation creates docking sites on the receptor for other molecules called STAT proteins (signal transducers and activators of transcription), and phosphorylated JAKs bind to a variety of STAT proteins. STAT proteins, or STATs, are DNA binding proteins that are activated by phosphorylation of tyrosine residues and function both as signaling molecules and transcription factors, and ultimately bind to specific DNA sequences present in the promoters of cytokine-responsive genes (Leonard et al, (2000), J.allergy Clin.Immunol.105: 877-888).

JAK/STAT signaling has been implicated in the modulation of many abnormal immune responses, such as allergy, asthma, autoimmune diseases such as transplant (allograft) rejection, rheumatoid arthritis, amyotrophic lateral sclerosis and multiple sclerosis, and in solid and hematologic cancers such as leukemia and lymphoma.

Thus, JAK and STAT are components of a number of potential cross-linked signal transduction pathways (Oncogene 19(2000), pp.5662-5679), suggesting the difficulty of specifically targeting one element of the JAK-STAT pathway without interfering with other signal transduction pathways.

JAK kinases, including JAK3, are abundantly expressed in primary leukemia cells of children with acute lymphoblastic leukemia, the most common form of childhood cancer, and studies have correlated STAT activation in certain cells with signals that regulate apoptosis (Demoulin et al, (1996), mol.cell.biol.16: 4710-6; Jurlander et al, (1997), blood.89: 4146-52; Kaneko et al, (1997), Clin.Exp.Immun.109: 185-193; and Nakamura et al, (1996), J.biol.chem.271: 19483-8). They are also known to be important for lymphocyte differentiation, function and survival. JAK3 plays a fundamental role in the function of lymphocytes, macrophages and mast cells, among others. Given the importance of this JAK kinase, compounds that modulate the JAK pathway, including those selective for JAK3, may be useful in the treatment of diseases or disorders involving lymphocyte, macrophage, or mast cell function (Kudlacz et al, (2004) am.j. transplantation 4: 51-57; Changelian (2003) Science 302: 875-. Conditions in which the JAK pathway is targeted or JAK kinases (particularly JAK3) are modulated are expected to be effective in the treatment of conditions including: leukemias, lymphomas, transplant rejection (e.g., islet transplant rejection, bone marrow transplant applications (e.g., graft versus host disease), autoimmune diseases (e.g., diabetes), and inflammation (e.g., asthma, allergy.) conditions that can benefit from inhibition of JAK3 are discussed in more detail below.

However, JAK3 has more restricted and regulated expression than the relatively ubiquitous expression of JAK1, JAK2, and Tyk 2. Although some JAKs (JAK1, JAK2, Tyk2) are used by various cytokine receptors, JAK3 is used only by cytokines that contain yc in their receptors. Therefore, JAK3 plays a role in cytokine signaling of cytokines, suggesting that cytokine receptors use the common γ chain to date; IL-2, IL-4, IL-7, IL-9, IL-15 and IL-21. Wherein JAK1 interacts with receptors for the cytokines IL-2, IL-4, IL-7, IL-9, and IL-21, and wherein JAK2 interacts with receptors for IL-9 and TNF- α. Receptor oligomerization occurs through the binding of certain cytokines to their receptors (e.g., IL-2, IL-4, IL-7, IL-9, IL-15, and IL-21), resulting in the proximity of the cytoplasmic tail of the bound JAK kinase and facilitating the transphosphorylation of tyrosine residues on the JAK kinase. This transphosphorylation leads to the activation of JAK kinases.

Animal experiments have indicated that JAK3 not only plays a key role in B and T lymphocyte maturation, but that JAK3 is constitutively required for maintaining T cell function. Modulation of immune activity by this novel mechanism can prove useful in the treatment of T cell proliferative disorders (e.g., transplant rejection) and autoimmune diseases.

In particular, JAK3 has been implicated in a variety of biological processes. For example, IL-4 and IL-9 induced proliferation and survival of murine mast cells has been shown to be dependent on JAK 3-and γ chain-signaling (Suzuki et al, (2000), Blood 96: 2172-. JAK3 also plays a key role in IgE receptor-mediated mast cell degranulation responses (Malaviya et al, (1999), biochem. Biophys. Res. Commun.257: 807. 813) and inhibition of JAK3 kinase has been shown to prevent type I hypersensitivity reactions (including anaphylaxis) (Malaviya et al, (1999), J.biol. chem.274: 27028. 27038). JAK3 was also shown to inhibit immunosuppression leading to allograft rejection (Kirken, (2001), Transpl. Proc.33: 3268-. JAK3 kinase is also implicated in mechanisms involved in the following diseases: early and late rheumatoid arthritis (Muller-Ladner et al, (2000), J.Immunal.164: 3894-3901); familial amyotrophic lateral sclerosis (Trieu et al, (2000), Biochem Biophys.Res.Commun.267: 22-25); leukemia (Sudbeck et al (1999), Clin. cancer Res.5: 1569-1582); mycosis fungoides, a form of T-cell lymphoma (Nielsen et al, (1997), Prac. Natl. Acad. Sci. USA 94: 6764-6769); and abnormal cell growth (Yu et al, (1997), J.Immunol.159: 5206-5210; Catlett-Falcone et al, (1999), Immunity 10: 105-115).

JAK3 inhibitors are useful as immunosuppressive agents for the effective treatment of organ transplantation, xenotransplantation, lupus, multiple sclerosis, rheumatoid arthritis, psoriasis, type I diabetes and complications from diabetes, cancer, asthma, atopic dermatitis, autoimmune thyroid disorders, ulcerative colitis, crohn's disease, alzheimer's disease, leukemia and other indications where immunosuppression is desirable.

Nonhematopoietic expression of JAK3 has also been reported, although its functional significance has yet to be elucidated (j. immunol.168(2002), pp.2475-2482). Since bone marrow transplantation for SCID is curative (Blood 103(2004), pp.2009-2018), JAK3 seems unlikely to have the necessary non-redundant functions in other tissues or organs. Therefore, in contrast to other targets for immunosuppressive drugs, a restricted distribution of JAK3 is desirable. Drugs that act on molecular targets with restricted expression in the immune system may result in optimal potency-to-toxicity ratios. Thus, targeting JAK3 would theoretically provide immunosuppression (i.e., act on cells actively involved in the immune response) when needed, without any effect outside of these cell populations. Albeit at various STATs^-/-Defective immune responses have been described in strains (J.Investig.Med.44(1996), pp.304-311; Curr.Opin.cell biol.9(1997), pp.233-239), but the ubiquitous distribution of STATs and the fact that those molecules lack enzymatic activity that can be targeted with small molecule inhibitors have made them an unmet candidate as a key target for immunosuppression.

SYK (spleen tyrosine kinase) is a non-receptor tyrosine kinase that is important for B cell activation via BCR signaling. SYK becomes activated upon binding to phosphorylated BCR and thus initiates early signaling events following BCR activation. SYK-deficient mice display an early block in B-cell development (Cheng et al Nature 378:303,1995; Turner et al Nature 378:298,1995). Therefore, inhibition of SYK enzymatic activity in cells has been suggested for the treatment of autoimmune diseases through its role in autoantibody production.

In addition to its role in BCR signaling and B cell activation, SYK also plays a key role in fceri-mediated mast cell degranulation and eosinophil activation. Thus, SYK is implicated in allergic disorders including asthma (reviewed in Wong et al Expert Opin investigatDrugs 13:743,2004). SYK binds via its SH2 domain to the phosphorylated gamma chain of fceri and is essential for downstream signaling (Taylor et al mol. SYK-deficient mast cells exhibit defective degranulation, arachidonic acid and cytokine secretion (Costello et al Oncogene 13:2595,1996). Pharmacological agents that inhibit SYK activity in mast cells are also shown (Yamamoto et al J Pharmacol ExpTher 306:1174,2003). Treatment with SYK antisense oligonucleotide drugs inhibited antigen-induced infiltration of eosinophils and neutrophils in an animal model of asthma (Stenton et al JImmunol 169:1028,2002). SYK-deficient eosinophils also show impaired activation in response to Fc ε R stimulation (Lach-Trifilefiffe et al Blood 96:2506,2000). Therefore, small molecule inhibitors of SYK would be useful for the treatment of allergy-induced inflammatory diseases including asthma.

In view of the wide variety of conditions that are expected to benefit from treatment involving modulation of the JAK and/or SYK pathway, it is apparent that novel compounds that modulate the JAK and/or SYK pathway and methods of using these compounds will provide substantial therapeutic benefit to a wide variety of patients. Provided herein are novel pyrrolopyrazine derivatives which are useful in the treatment of disorders directed to the JAK and/or SYK pathway or inhibiting JAK and/or SYK kinases, particularly JAK3, and which are therapeutically effective in the treatment of autoimmune and inflammatory diseases.

The novel pyrrolopyrazine derivatives provided herein selectively inhibit JAK3 and are useful in the treatment of autoimmune and inflammatory diseases. The compounds of the present invention modulate the JAK and/or SYK pathway and are novel pyrrolopyrazine derivatives useful in the treatment of autoimmune and inflammatory diseases, with preferred compounds selectively inhibiting JAK 3. For example, the compounds of the present invention may inhibit JAK3 and SYK, with preferred compounds being selective for JAK3 in JAK kinases and being novel pyrrolopyrazine derivatives useful in the treatment of autoimmune and inflammatory diseases. Furthermore, the compounds of the present invention can inhibit JAK3 and JAK2, with preferred compounds being selective for JAK3 among JAK kinases and being novel pyrrolopyrazine derivatives useful in the treatment of autoimmune and inflammatory diseases. Similarly, the compounds of the present invention may inhibit JAK3 and JAK1, with preferred compounds being selective for JAK3 in the JAK kinases and being novel pyrrolopyrazine derivatives useful in the treatment of autoimmune and inflammatory diseases.

The present application provides compounds of formula I or a pharmaceutically acceptable salt thereof,

wherein:

y is C (R)¹)₂(C(R¹’)₂)_m

m is 0 or 1;

R¹each is H or R^1a；

R^1aEach independently is lower alkyl, lower alkoxy, phenyl, benzyl, heteroaryl, cycloalkyl, heterocycloalkyl, or cycloalkyl lower alkyl, optionally substituted with one or more R^1a' substitution;

R^1a' is halogen, lower alkyl, lower haloalkyl, lower alkoxy, lower hydroxyalkyl, oxo, hydroxy or cyano;

R¹' are each independently H, lower alkyl or lower haloalkyl;

R²independently is H or R^2a；

R^2aIndependently is lower alkyl, lower haloalkyl, lower alkoxy, lower hydroxyalkyl, cyano lower alkyl, cycloalkyl or heterocycloalkyl;

or R^2aAnd R^1aTogether form a ring, which is optionally substituted with one or more of the following groups: halogen, lower alkyl, cyano lower alkyl, hydroxy, lower haloalkyl, lower hydroxyAlkyl, lower alkoxy, lower alkylamino or lower dialkylamino;

R³independently is H or R^3a；

R^3aIndependently is lower alkyl, lower haloalkyl, lower alkoxy, lower hydroxyalkyl, cyano lower alkyl, C (= O) R^3a' or S (= O)₂R^3a’；

R^3a' are each independently H or lower alkyl;

q is H, halogen, hydroxy, cyano or Q';

q' is lower alkyl, lower alkenyl, lower alkynyl, cycloalkyl, phenyl, cycloalkenyl, heterocycloalkyl or heteroaryl, optionally substituted with one or more Q^aSubstitution;

Q^ais Q^bOr Q^c；

Q^bIs halogen, oxo, hydroxy, -CN, -SCH₃、–S(O)₂CH₃or-S (= O) CH₃；

Q^cIs Q^dOr Q^e；

Or two Q^aTogether form a bicyclic ring system, optionally substituted with one or more Q^bOr Q^cSubstitution;

Q^dis-O (Q)^e)、–S(=O)₂(Q^e)、–C(=O)N(Q^e)₂、–S(O)₂(Q^e)、–C(=O)(Q^e)、–C(=O)O(Q^e)、–N(Q^e)₂；–N(Q^e)C(=O)(Q^e)、–N(Q^e)C(=O)O(Q^e) or-N (Q)^e)C(=O)N(Q^e)₂；

Q^eEach independently is H or Q^e’；

Q^e' each independently is a lower alkanePhenyl, benzyl, lower haloalkyl, lower alkoxy, cycloalkyl, cycloalkenyl, heterocycloalkyl or heteroaryl, optionally substituted with one or more Q^fSubstitution;

Q^fis Q^gOr Q^h；

Q^gIs halogen, hydroxy, cyano, oxo or-C (= O) (Q)^h)；

Q^hIs lower alkyl, lower haloalkyl, lower alkoxy, amino, phenyl, benzyl, cycloalkyl, heterocycloalkyl or heteroaryl, optionally substituted with one or more QⁱSubstitution; and is

QⁱIs halogen, hydroxy, cyano, lower alkyl, lower haloalkyl or lower alkoxy.

In one aspect, the present application provides a method of treating an inflammatory and/or autoimmune disease comprising administering to a patient in need thereof a therapeutically effective amount of a compound of formula I.

The present application provides a pharmaceutical composition comprising a compound of formula I in admixture with at least 1 pharmaceutically acceptable carrier, excipient or diluent.

Definition of

The term "an" entity as used herein refers to one or more of that entity; for example, a compound refers to one or more compounds or at least one compound. Thus, the terms "a", "an", "one or more" and "at least one" are used interchangeably herein.

The term "as defined above" refers to the broadest definition or broadest claim of each group provided in the summary of the invention. In all other embodiments provided below, substituents that can be present in each embodiment and are not explicitly defined retain the broadest definition provided in the summary of the invention.

As used in this specification, the terms "comprises" and "comprising," whether in transitional terms or in the claims, are to be construed as having an open-ended meaning. That is, the term is to be interpreted as having the same meaning as the term "having at least" or "including at least". When used in the context of a method, the term "comprising" means that the method includes at least the recited steps, but may include other steps. The term "comprising" when used in the context of a compound or composition means that the compound or composition includes at least the recited features or components, but may also include other features or components.

As used herein, unless otherwise specifically indicated, the word "or" is used in the "inclusive" sense of "and/or" and not in the "exclusive" sense of "or".

The term "independently" as used herein means that the variables apply in either case, regardless of whether variables having the same or different definitions are present in the same compound. Thus, in a compound where R "appears 2 times and is defined as" independently being carbon or nitrogen ", 2R" can all be carbon, 2R "can all be nitrogen, or one R" can be carbon and the other nitrogen.

When any variable (e.g., R, R' or Q) occurs more than 1 time in any moiety or formula describing a compound used or claimed in the present invention, its definition in each case is independent of the other. Moreover, combinations of substituents and/or variables are permissible only if such compounds result in stable compounds.

The symbol "+" at the end of a bond or "- - - -" through a bond represents the point at which a functional group or other chemical moiety is attached to the rest of the molecule. Thus, for example

MeC(=O)OR⁴Wherein

A bond inserted into a ring system (which is different from the bond at the different end points) means that the bond can be attached to any suitable ring atom.

The term "optional" or "optionally" as used herein means that the subsequently described event or circumstance may, but need not, occur, and that the statement includes instances where the event or circumstance occurs and instances where it does not. For example, "optionally substituted" means that the optionally substituted moiety may represent hydrogen or a substituent.

The term "together form a bicyclic ring system" as used herein means linked to form a bicyclic ring system, wherein each ring may be composed of 4 to 7 carbon atoms or 4 to 7 carbons and heteroatoms, and may be saturated or unsaturated.

The term "together form a ring" as used herein means that the ring is formed together with the atoms to which the substituents are attached, wherein the ring may consist of 4 to 7 carbon atoms or 4 to 7 carbon atoms and heteroatoms, and may be saturated or unsaturated.

The term "about" as used herein means approximately, to the left of, roughly, or around. When the term "about" is used in connection with a numerical range, it modifies that range by extending the upper and lower bounds of the numerical values set forth. Generally, the term "about" is used herein to refer to a numerical value that differs by 20% from the upper or lower stated value.

The definitions described herein may be combined to form chemically relevant combinations such as "heteroalkylaryl," "haloalkylheteroaryl," "arylalkyl heterocyclyl," "alkylcarbonyl," "alkoxyalkyl," "cycloalkylalkyl," and the like. When the term "alkyl" is used as a suffix following another term, as in "phenylalkyl" or "hydroxyalkyl", this refers to an alkyl group, as defined below, which is substituted with 1-2 substituents selected from another group specifically named. Thus, for example, "phenylalkyl" refers to an alkyl group having 1-2 phenyl substituents, and thus includes benzyl, phenethyl, and biphenyl. "alkylaminoalkyl" is an alkyl group having 1-2 alkylamino substituents. "hydroxyalkyl" includes 2-hydroxyethyl, 2-hydroxypropyl, 1- (hydroxymethyl) -2-methylpropyl, 2-hydroxybutyl, 2, 3-dihydroxybutyl, 2- (hydroxymethyl), 3-hydroxypropyl and the like. Thus, as used herein, the term "hydroxyalkyl" is used to define a subset of heteroalkyl groups defined below. The term- (ar) alkyl refers to unsubstituted alkyl or aralkyl. The term (hetero) aryl refers to an aryl or heteroaryl group.

The compounds of formula I may exhibit tautomerism. Tautomeric compounds can exist in 2 or more tautomeric species. Migration of covalently bound hydrogen atoms between 2 atoms results in prototropic tautomers. Tautomers are usually in equilibrium, and attempts to separate individual tautomers usually result in mixtures whose chemical and physical properties are consistent with a mixture of compounds. The equilibrium point depends on the chemical nature within the molecule. For example, in many aliphatic aldehydes and ketones, such as acetaldehyde, the ketoform predominates; whereas in phenols the enol form predominates. Common prototropic tautomers include keto/enol (-C (= O) -CH-and-C (-OH) = CH-), amide/imide (-C (= O) -NH-and-C (-OH) = N-) and amidine (-C (= NR) -NH-and-C (-NHR) = N-) tautomers. The latter 2 species are particularly common in heteroaryl and heterocyclic rings, and the present invention includes all tautomeric forms of the compounds.

Unless defined otherwise, technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Reference is made herein to various methodologies and literature materials known to those skilled in the art. Standard references which set forth general principles of pharmacology include The Pharmacological Basis of Therapeutics, 10 th edition, McGrawHill Companies Inc., New York (2001), by Goodman and Gilman. Any suitable materials and/or methods known to the skilled person can be used in the practice of the present invention. However, preferred materials and methods are described. Unless otherwise indicated, materials, reagents and the like referred to in the following specification and examples may be obtained from commercial sources.

The term "acyl" as used herein refers to an acyl group of the formula-A group of C (= O) R, wherein R is hydrogen or lower alkyl as defined herein. The term or "alkylcarbonyl" as used herein refers to a group of formula C (= O) R, wherein R is alkyl as defined herein. Term C_1-6Acyl means that the-C (= O) R group contains 6 carbon atoms. The term "arylcarbonyl" as used herein refers to a group of formula C (= O) R, wherein R is an aryl group; the term "benzoyl" as used herein refers to an "arylcarbonyl" group wherein R is phenyl.

The term "alkyl" as used herein refers to an unbranched or branched chain-like, saturated, monovalent hydrocarbon radical containing from 1 to 10 carbon atoms. The term "lower alkyl" refers to a straight or branched chain hydrocarbon group containing 1 to 6 carbon atoms. As used herein, "C_1-10Alkyl "refers to an alkyl group consisting of 1-10 carbons. Examples of alkyl groups include, without limitation, lower alkyl groups including methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, tert-butyl or pentyl, isopentyl, neopentyl, hexyl, heptyl, and octyl.

When the term "alkyl" is used as a suffix following another term, as in "phenylalkyl" or "hydroxyalkyl", this refers to an alkyl group as defined above substituted with 1 to 2 substituents selected from other explicitly named groups. Thus, for example, "phenylalkyl" refers to an R 'R "-group, wherein R' is a phenyl group, and R" is an alkylene group, as defined herein, it being understood that the point of attachment of the phenylalkyl moiety will be on the alkylene group. Examples of arylalkyl groups include, but are not limited to, benzyl, phenethyl, 3-phenylpropyl. The term "arylalkyl" or "aralkyl" is similarly construed, wherein R' is aryl. The term "heteroarylalkyl" is similarly interpreted, wherein R' is optionally aryl or heteroaryl.

The term "haloalkyl" as used herein refers to an unbranched or branched chain alkyl group as defined above wherein 1,2, 3 or more hydrogen atoms are replaced by a halogen. The term "lower haloalkyl" refers to a straight or branched chain hydrocarbon residue containing 1 to 6 carbon atoms, wherein 1,2, 3 or more hydrogen atoms are substituted with halogen. Examples are 1-fluoromethyl, 1-chloromethyl, 1-bromomethyl, 1-iodomethyl, difluoromethyl, trifluoromethyl, trichloromethyl, tribromomethyl, triiodomethyl, 1-fluoroethyl, 1-chloroethyl, 1-bromoethyl, 1-iodoethyl, 2-fluoroethyl, 2-chloroethyl, 2-bromoethyl, 2-iodoethyl, 2, 2-dichloroethyl, 3-bromopropyl or 2,2, 2-trifluoroethyl.

The term "alkylene" as used herein, unless otherwise specified, refers to a divalent saturated straight chain hydrocarbon radical of 1 to 10 carbon atoms (e.g., (CH)₂)_n) Or a branched, saturated, divalent hydrocarbon radical of 2 to 10 carbon atoms (e.g. -CHMe-or-CH)₂CH(i-Pr)CH₂-). Except in the case of methylene, the open valences of the alkylene groups are not attached to the same atom. Examples of alkylene include, without limitation, methylene, ethylene, propylene, 2-methyl-propylene, 1-dimethyl-ethylene, butylene, 2-ethylbutylene.

The term "alkoxy" as used herein refers to an-O-alkyl group wherein alkyl is as defined above, e.g., methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, t-butoxy, pentyloxy, hexyloxy, including isomers thereof. "lower alkoxy" as used herein refers to an alkoxy group having a "lower alkyl" group as previously defined. As used herein, "C_1-10Alkoxy "means an-O-alkyl group wherein alkyl is C_1-10。

The term "hydroxyalkyl" as used herein refers to an alkyl group as defined herein wherein 1 to 3 hydrogen atoms on different carbon atoms are replaced by a hydroxyl group.

The term "cycloalkyl" as used herein refers to a saturated carbocyclic ring containing from 3 to 8 carbon atoms, i.e. cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl or cyclooctyl. As used herein, "C_3-7Cycloalkyl "refers to a cycloalkyl group consisting of 3 to 7 carbons in a carbocyclic ring.

Unless otherwise indicated, the term "cycloalkenyl" refers to partially unsaturated groups containing from 5 to 7 carbon atomsCarbocyclic, and having a carbon-carbon double bond in the ring. E.g. C_5-6Cycloalkenyl refers to cycloalkenyl groups having from 5 to 6 carbon atoms. In certain embodiments, cycloalkenyl groups have one carbon-carbon double bond in the ring. In certain embodiments, cycloalkenyl groups have multiple carbon-carbon double bonds in the ring. However, the cycloalkenyl ring is not aromatic. Cycloalkenyl groups may be optionally substituted with one or more substituents. Examples of cycloalkenyl groups include, but are not limited to, cyclopentenyl and cyclohexenyl.

The term "halogen" as used herein refers to fluorine, chlorine, bromine or iodine.

The term "amino" as used herein includes-NR₂Wherein each R group is independently H or lower alkyl, wherein lower alkyl is as defined herein. Examples of amino groups include dimethylamino, methylamino and NH₂。

The term "aryl" as used herein refers to a monocyclic or bicyclic (also referred to as "biaryl") substituted or unsubstituted carbocyclic aromatic group. Examples of aryl groups are phenyl, naphthyl, and the like.

The term "heteroaryl" as used herein refers to a monocyclic, bicyclic (heterodiaryl) or tricyclic group of 5 to 18 ring atoms having at least one aromatic ring, each ring containing 4 to 8 atoms, including one or more N, O or S heteroatoms, the other ring atoms being carbon, it being understood that the attachment point of the heteroaryl group will be on the aromatic ring. As is well known to those skilled in the art, heteroaryl rings have less aromaticity than their full carbon counterparts. Thus, for the purposes of the present invention, heteroaryl groups need only have some degree of aromaticity. Examples of heteroaryl moieties include monocyclic aromatic heterocycles having 5-6 ring atoms and 1-3 heteroatoms, including, without limitation, pyridyl, pyrimidinyl, pyrazinyl, pyrrolyl, pyrazolyl, imidazolyl, indolyl, substituted phenyl, pyridyl, pyrazin,Azolyl radical, isoOxazolyl, thiazole, isothiazole, triazoline, triazolyl, thienyl, furyl, thiadiazole anda bisoxazoline which may be optionally substituted with one or more, preferably 1 or 2, substituents selected from the group consisting of hydroxy, cyano, alkyl, alkoxy, thio, lower haloalkoxy, alkylthio, halogen, haloalkyl, alkylsulfinyl, alkylsulfonyl, halogen, amino, alkylamino, dialkylamino, aminoalkyl, alkylaminoalkyl and dialkylaminoalkyl, nitro, alkoxycarbonyl and carbamoyl, alkylcarbamoyl, dialkylcarbamoyl, arylcarbamoyl, alkylcarbonylamino and arylcarbonylamino. Examples of bicyclic moieties include, without limitation, quinolinyl, indazolyl, isoquinolinyl, benzofuranyl, benzothienyl, benzoAzole, benzisohOxazole, benzothiazole, pyrrolopyridinyl, pyrrolopyrazinyl and benzisothiazole.

The terms "heterocycloalkyl", "heterocyclyl" or "heterocycle" as used herein, unless otherwise specified, refer to a monovalent saturated cyclic radical consisting of one or more rings, preferably 1-2 or 3 rings, each ring having 3-8 atoms, containing one or more ring carbon atoms and one or more ring heteroatoms (selected from N, O or S (= O)_0-2) Wherein the point of attachment can be through a carbon or heteroatom and which can optionally be independently substituted with one or more, preferably 1 or 2 or 3, substituents selected from hydroxy, oxo, cyano, lower alkyl, lower alkoxy, lower haloalkoxy, alkylthio, halogen, haloalkyl, hydroxyalkyl, nitro, alkoxycarbonyl, amino, alkylamino, alkylsulfonyl, arylsulfonyl, alkylaminosulfonyl, optionally substituted with one or more, preferably 1 or 2 or 3, substituents selected from the group consisting of hydroxy, oxo, cyano, lower alkyl, lower alkoxy, lower haloalkoxy, alkylthio, halogen, haloalkyl, hydroxyalkyl, nitro, alkoxycarbonyl, amino, alkylamino, alkylsulfonyl, arylsulfonyl,arylaminosulfonyl, alkylsulfonylamino, arylsulfonylamino, alkylaminocarbonyl, arylaminocarbonyl, alkylcarbonylamino, arylcarbonylamino. Examples of heterocyclic groups include, without limitation, azetidinyl, pyrrolidinyl, hexahydroazepinylAn oxacyclobutyl group, a tetrahydrofuryl group, a tetrahydrothienyl group,Oxazolidinyl, thiazolidinyl, isooxazolidinylOxazolidinyl, pyrrolidinyl, morpholinyl, piperazinyl, piperidinyl, isoindolinyl, dihydroisoquinolinyl, tetrahydropyranyl, tetrahydrocarbolinyl, imidazolinyl, thiomorpholinyl, quinuclidinyl, and imidazolinyl groups.

The term "organ rejection" includes acute allograft or xenograft rejection and chronic allograft or xenograft rejection in the case of vascularized and/or non-vascularized (e.g. bone marrow, islet cells) transplants.

JAK inhibitors

The present application provides compounds of formula I or pharmaceutically acceptable salts thereof

Wherein:

y is C (R)¹)₂(C(R¹’)₂)_m

m is 0 or 1;

R¹each is H or R^1a；

R^1aEach is independentIs lower alkyl, lower alkoxy, phenyl, benzyl, heteroaryl, cycloalkyl, heterocycloalkyl or cycloalkyl lower alkyl, optionally substituted with one or more R^1a' substitution;

R¹' are each independently H, lower alkyl or lower haloalkyl;

R²independently is H or R^2a；

R³independently is H or R^3a；

R^3a' are each independently H or lower alkyl;

q is H, halogen, hydroxy, cyano or Q';

Q^ais Q^bOr Q^c；

Q^bIs halogen, oxo, hydroxy, -CN, -SCH₃、–S(O)₂CH₃or-S (= O) CH₃；

Q^cIs Q^dOr Q^e；

Q^eEach independently is H or Q^e’；

Q^e' are each independently lower alkyl, phenyl, benzyl, lower haloalkyl, lower alkoxy, cycloalkyl, cycloalkenyl, heterocycloalkyl, or heteroaryl, optionally substituted with one or more Q^fSubstitution;

Q^fis Q^gOr Q^h；

Q^gIs halogen, hydroxy, cyano, oxo or-C (= O) (Q)^h)；

QⁱIs halogen, hydroxy, cyano, lower alkyl, lower haloalkyl or lower alkoxy.

The present application provides compounds of formula I as described above, wherein Q is cycloalkyl or heterocycloalkyl, optionally substituted with one or more Q^aAnd (4) substitution.

The present application provides compounds of formula I as described above, wherein Q is cycloalkyl.

The present application provides compounds of formula I as described above, wherein Q is cyclopropyl, optionally substituted with one or more Q^aAnd (4) substitution.

The application provides compounds of formula I, wherein one R¹Is H, lower alkyl or cycloalkyl or R^2aAnd R¹Together form a piperidinyl ring, and the other R¹Is H.

The application provides compounds of formula I, wherein one R¹Is lower alkyl or cycloalkyl, and other R¹Is H.

The application provides compounds of formula I, wherein one R¹Is lower alkyl, and the other R¹Is H.

The present application provides compounds of formula I, wherein Q is cyclopropyl, optionally substituted with one or more Q^aSubstituted, one R¹Is lower alkyl, and the other R¹Is H.

The present application provides compounds of formula I, wherein m is 0.

The present application provides compounds of formula I, wherein Q is cyclopropyl, optionally substituted with one or more Q^aAnd m is 0.

The application provides compounds of formula I, R¹Each independently is H, lower alkyl or cycloalkyl.

The present application provides compounds of formula I, Q is cyclopropyl, optionally substituted with one or more Q^aIs substituted, and R¹Each independently is H, lower alkyl or cycloalkyl.

The present application provides compounds of formula I, wherein R¹Is methyl, cyclopropyl or sec-butyl.

The present application provides compounds of formula I, wherein R¹Is methyl, cyclopropyl or sec-butyl, Q is cyclopropyl, which is optionallyBy one or more Q^aIs substituted, and R¹Each independently is H, lower alkyl or cycloalkyl.

The present application provides compounds of formula I, wherein R^2aAnd R^1aTogether form a ring, which is optionally substituted with lower alkyl, cyano or cyano lower alkyl.

The present application provides compounds of formula I, wherein Q is cyclopropyl, optionally substituted with one or more Q^aSubstituted, m is 0, and R^2aAnd R^1aTogether form a ring, which is optionally substituted with lower alkyl, cyano or cyano lower alkyl.

The application provides compounds of formula I, wherein m is 1 and R¹' are each H.

The present application provides compounds of formula I, wherein Q is cyclopropyl, optionally substituted with one or more Q^aSubstituted, m is 1 and R¹' are each H.

The present application provides compounds of formula I, wherein R¹Each independently is H, lower alkyl or cycloalkyl, Q is cyclopropyl, optionally substituted with one or more Q^aSubstituted, m is 1 and R¹' are each H.

The present application provides compounds of formula I, wherein R²And R³Independently is H, lower alkyl, cycloalkyl, cyano lower alkyl or lower haloalkyl.

The present application provides compounds of formula I, wherein R²And R³Independently is H, lower alkyl, cycloalkyl, cyano lower alkyl or lower haloalkyl, and Q is cyclopropyl, optionally substituted with one or more Q^aAnd (4) substitution.

The present application provides compounds of formula I, wherein R²And R³Independently is H, lower alkylCycloalkyl, cyano lower alkyl or lower haloalkyl, Q is cyclopropyl, optionally substituted with one or more Q^aAnd m is 0.

The present application provides compounds of formula I, wherein Q is cyclopropyl, optionally substituted with one or more Q^aSubstituted, m is 1, R¹' each is H, and R²And R³Independently is H, lower alkyl, cycloalkyl, cyano lower alkyl or lower haloalkyl.

The present application provides a compound selected from the group consisting of:

2-cyclopropyl-5H-pyrrolo [2,3-b ] pyrazine-7-carboxylic acid carbamoylmethyl-amide;

2-cyclopropyl-5H-pyrrolo [2,3-b ] pyrazine-7-carboxylic acid [ 1-methyl-2- (2,2, 2-trifluoro-ethylcarbamoyl) -ethyl ] -amide;

2-cyclopropyl-5H-pyrrolo [2,3-b ] pyrazine-7-carboxylic acid (2-ethylcarbamoyl-1-methyl-ethyl) -amide;

2-cyclopropyl-5H-pyrrolo [2,3-b ] pyrazine-7-carboxylic acid (1-methyl-2-propylcarbamoyl-ethyl) -amide;

2-cyclopropyl-5H-pyrrolo [2,3-b ] pyrazine-7-carboxylic acid (2-cyclopropylcarbamoyl-1-methyl-ethyl) -amide;

2-cyclopropyl-5H-pyrrolo [2,3-b ] pyrazine-7-carboxylic acid ((R) -1-dimethylcarbamoyl-ethyl) -amide;

2-cyclopropyl-5H-pyrrolo [2,3-b ] pyrazine-7-carboxylic acid (2-carbamoyl-1-cyclopropyl-2-methyl-propyl) -amide;

2-cyclopropyl-5H-pyrrolo [2,3-b ] pyrazine-7-carboxylic acid [ (R) -1- (cyanomethyl-methyl-carbamoyl) -ethyl ] -amide;

2-cyclopropyl-5H-pyrrolo [2,3-b ] pyrazine-7-carboxylic acid ((S) -1-dimethylcarbamoyl-ethyl) -amide; and

2-cyclopropyl-5H-pyrrolo [2,3-b ] pyrazine-7-carboxylic acid ((R) -1-dimethylcarbamoyl-3-methyl-butyl) -amide.

2-cyclopropyl-5H-pyrrolo [2,3-b ] pyrazine-7-carboxylic acid ((R) -2-oxo-piperidin-3-yl) -amide;

2-cyclopropyl-5H-pyrrolo [2,3-b ] pyrazine-7-carboxylic acid ((R) -1-methyl-2-oxo-piperidin-3-yl) -amide; and

2-cyclopropyl-5H-pyrrolo [2,3-b ] pyrazine-7-carboxylic acid [ (R) -1- (2-cyano-ethyl) -2-oxo-piperidin-3-yl ] -amide.

The present application provides a method of treating an inflammatory or autoimmune disease comprising administering to a patient in need thereof a therapeutically effective amount of a compound of formula I.

The present application provides the above method further comprising administering an additional therapeutic agent selected from a chemotherapeutic or anti-proliferative agent, an anti-inflammatory agent, an immunomodulatory or immunosuppressive agent, a neurotrophic factor, an agent for treating cardiovascular disease, an agent for treating diabetes, or an agent for treating immunodeficiency disorders.

The present application provides a method of treating rheumatoid arthritis comprising administering to a patient in need thereof a therapeutically effective amount of a compound of formula I.

The present application provides a method of treating asthma comprising administering to a patient in need thereof a therapeutically effective amount of a compound of formula I.

The present application provides a method of treating an inflammatory disorder comprising administering to a patient in need thereof a therapeutically effective amount of a compound of formula I.

The present application provides a method of inhibiting a T-cell proliferative disease comprising administering to a patient in need thereof a therapeutically effective amount of a compound of formula I.

The present application provides the above method wherein the proliferative disease is cancer.

The present application provides a method of treating a B-cell proliferative disease comprising administering to a patient in need thereof a therapeutically effective amount of a compound of formula I.

The present application provides methods of treating immune diseases including lupus, multiple sclerosis, rheumatoid arthritis, psoriasis, type I diabetes, organ transplant complications, xenotransplantation, diabetes, cancer, asthma, atopic dermatitis, autoimmune thyroid disease, ulcerative colitis, crohn's disease, alzheimer's disease, and leukemia comprising administering to a patient in need thereof a therapeutically effective amount of a compound of formula I.

The present application provides methods of preventing or treating all forms of organ rejection, including acute allograft or xenograft rejection and chronic allograft or xenograft rejection of vascularized or non-vascularized transplants, comprising administering a compound of formula I to a patient in need thereof.

The present application provides a method of inhibiting JAK3 activity comprising administering a compound of formula I, wherein the compound exhibits an IC of 50 micromolar or less in an in vitro biochemical assay of JAK3 activity₅₀。

The present application provides the above-described methods wherein the compound exhibits an IC of 100 nanomolar or less in an in vitro biochemical assay for JAK3 activity₅₀。

The present application provides the above-described methods wherein the compound exhibits an IC of 10 nanomolar or less in an in vitro biochemical assay for JAK3 activity₅₀。

A method of inhibiting SYK activity comprising administering a compound of formula I, wherein the compound exhibits an IC of 50 micromolar or less in an in vitro biochemical assay for SYK activity₅₀。

The present application provides the above-described method, wherein the compound exhibits an IC of 100 nanomolar or less in an in vitro biochemical assay of SYK activity₅₀。

The present application provides the above-described method, wherein the compound exhibits an IC of 10 nanomolar or less in an in vitro biochemical assay of SYK activity₅₀。

The present application provides a method of treating an inflammatory disorder comprising administering to a patient in need thereof a therapeutically effective amount of an anti-inflammatory compound in combination with a compound of formula I.

The present application provides a method of treating an immune disease comprising administering to a patient in need thereof a therapeutically effective amount of an immunosuppressive compound in combination with a compound of formula I.

The present application provides the above composition further comprising an additional therapeutic agent selected from the group consisting of chemotherapeutic or anti-proliferative agents, anti-inflammatory agents, immunomodulatory or immunosuppressive agents, neurotrophic factors, agents for treating cardiovascular disease, agents for treating diabetes, and agents for treating immunodeficiency disorders.

The present application provides processes for preparing compounds of formula I.

The present application provides the compounds described above for use in the treatment of inflammatory or autoimmune diseases.

The present application provides the compounds described above for use in the treatment of any one of the diseases described above.

The present application provides the use of a compound of formula I for the manufacture of a medicament for the treatment of inflammatory diseases.

The present application provides the use of a compound of formula I for the preparation of a medicament for the treatment of an autoimmune disease.

The present application provides the invention as described above.

The present application provides compounds of formula I' or pharmaceutically acceptable salts thereof

Wherein:

y is C (R)¹)₂(C(R¹’)₂)_m

m is 0 or 1;

R¹each is H or R^1a；

R¹' are each independently H, lower alkyl or lower haloalkyl;

R²independently is H or R^2a；

or R^2aAnd R^1aTogether form a ring, which is optionally substituted with one or more of the following groups: halogen, lower alkyl, cyano lower alkyl, hydroxy, lower haloalkyl,Lower hydroxyalkyl, lower alkoxy, lower alkylamino or lower dialkylamino;

R³independently is H or R^3a；

R^3a' are each independently H or lower alkyl;

q is H, halogen, hydroxy, cyano or Q';

Q^ais Q^bOr Q^c；

Q^bIs halogen, oxo, hydroxy, -CN, -SCH₃、–S(O)₂CH₃or-S (= O) CH₃；

Q^cIs Q^dOr Q^e；

Q^eEach independently is H or Q^e’；

Q^e' independently of each otherIs lower alkyl, phenyl, benzyl, lower haloalkyl, lower alkoxy, cycloalkyl, cycloalkenyl, heterocycloalkyl or heteroaryl, optionally substituted with one or more Q^fSubstitution;

Q^fis Q^gOr Q^h；

Q^gIs halogen, hydroxy, cyano, oxo or-C (= O) (Q)^h)；

QⁱIs halogen, hydroxy, cyano, lower alkyl, lower haloalkyl or lower alkoxy.

In a variant of formula I', R¹Is H.

In a variant of formula I', R¹Is R^1a。

In one variation of formula I', Q is cycloalkyl or heterocycloalkyl, optionally substituted with one or more R^1a' substitution.

In one variation of formula I', Q is cycloalkyl, optionally substituted with one or more R^1a' substitution.

In one variation of formula I', Q is cyclopropyl, optionally substituted with one or more R^1a' substitution.

In a variant of formula I', R^1a' is lower alkyl.

In one variation of formula I', Q is cyclopropyl.

In one variation of formula I', Q is heterocycloalkyl, optionally substituted with one or more R^1a' substitution.

In one variation of formula I', Q is pyrazolyl.

In one variation of formula I', m is 0.

In a variant of formula I', R¹Each independently is H, lower alkyl or cycloalkyl.

In a variant of formula I', Y is CHR^1a。

In a variant of formula I', R^1aIs lower alkyl or cycloalkyl.

In a variant of formula I', R^1aIs methyl, cyclopropyl or sec-butyl.

In a variant of formula I', R^2aAnd R^1aTogether form a ring, which is optionally substituted with lower alkyl, cyano or cyano lower alkyl.

In a variant of formula I', R^2aAnd R^1aTogether forming a ring.

In one variation of formula I', m is 1.

In a variant of formula I', R²And R³Independently is H, lower alkyl, cycloalkyl, cyano lower alkyl or lower haloalkyl.

In a variant of formula I', R²Or R³Is H.

2-cyclopropyl-5H-pyrrolo [2,3-b ] pyrazine-7-carboxylic acid ((S) -1-dimethylcarbamoyl-ethyl) -amide;

2-cyclopropyl-5H-pyrrolo [2,3-b ] pyrazine-7-carboxylic acid ((R) -1-dimethylcarbamoyl-3-methyl-butyl) -amide;

In one variation of the above method, the above method further comprises administering an additional therapeutic agent selected from a chemotherapeutic or anti-proliferative agent, an anti-inflammatory agent, an immunomodulatory or immunosuppressive agent, a neurotrophic factor, an agent for treating cardiovascular disease, an agent for treating diabetes, or an agent for treating immunodeficiency disorders.

In one aspect, the application provides a method of treating an inflammatory disorder comprising administering to a patient in need thereof a therapeutically effective amount of a compound of formula I.

In one aspect, the application provides a method of inhibiting a T-cell proliferative disease comprising administering to a patient in need thereof a therapeutically effective amount of a compound of formula I.

In one variation of the above method, the proliferative disease is cancer.

In one aspect, the application provides a method of treating a B-cell proliferative disease comprising administering to a patient in need thereof a therapeutically effective amount of a compound of formula I.

In one aspect, the application provides a method of treating an immune disease comprising administering to a patient in need thereof a therapeutically effective amount of a compound of formula I, said immune disease comprising lupus, multiple sclerosis, rheumatoid arthritis, psoriasis, type I diabetes, organ transplant complications, xenotransplantation, diabetes, cancer, asthma, atopic dermatitis, autoimmune thyroid disease, ulcerative colitis, crohn's disease, alzheimer's disease, and leukemia.

In one aspect, the present application provides a method of preventing or treating all forms of organ rejection, including acute allograft or xenograft rejection and chronic allograft or xenograft rejection of vascularized or non-vascularized transplants, comprising administering to a patient in need thereof a therapeutically effective amount of a compound of formula I.

In one aspect, the application provides a method of inhibiting JAK3 activity comprising administering a compound of formula I', wherein the compound exhibits an IC of 50 micromolar or less in an in vitro biochemical assay of JAK3 activity₅₀。

In a variation of the above method, wherein the compound exhibits an IC of 100 nanomolar or less in an in vitro biochemical assay for JAK3 activity₅₀。

In a variation of the above method, wherein the compound exhibits an IC of 10 nanomolar or less in an in vitro biochemical assay for JAK3 activity₅₀。

In one aspect, the present application provides a method of inhibiting SYK activity comprising administering a compound of formula I, wherein the compound exhibits an IC of 50 micromolar or less in an in vitro biochemical assay of SYK activity₅₀。

In a variation of the above method, wherein the compound exhibits an IC of 100 nanomolar or less in an in vitro biochemical assay of SYK activity₅₀。

In a variation of the above method, wherein the compound exhibits an IC of 10 nanomolar or less in an in vitro biochemical assay of SYK activity₅₀。

In one aspect, the application provides a method of treating an inflammatory disorder comprising administering to a patient in need thereof an anti-inflammatory compound in combination with a therapeutically effective amount of a compound of formula I.

In one aspect, the present application provides a method of treating an immune disease comprising administering to a patient in need thereof an immunosuppressive compound in combination with a therapeutically effective amount of a compound of formula I.

The present application provides a pharmaceutical composition comprising a compound of formula I' in admixture with at least 1 pharmaceutically acceptable carrier, excipient or diluent.

In a variation, the pharmaceutical composition described above further comprises an additional therapeutic agent selected from the group consisting of chemotherapeutic or anti-proliferative agents, anti-inflammatory agents, immunomodulatory or immunosuppressive agents, neurotrophic factors, agents for treating cardiovascular disease, agents for treating diabetes, and agents for treating immunodeficiency disorders.

In one aspect, the application provides the use of a compound of formula I in the manufacture of a medicament for the treatment of an inflammatory disease.

In one aspect, the application provides the use of a compound of formula I for the manufacture of a medicament for the treatment of an autoimmune disease.

The present application provides a compound or method as described herein.

Compound (I)

Examples of representative compounds encompassed by and within the scope of the present invention are provided in the following tables. The following examples and preparations are provided to enable those skilled in the art to more clearly understand and practice the present invention. They should not be considered as limiting the scope of the invention, but merely as being illustrative and representative thereof.

Generally, the nomenclature used in this application is based on AUTONOMTM v.4.0, the Beilstein Institute computerized system used to generate IUPAC systematic nomenclature. The depicted structure is meant to control if there is a discrepancy between the depicted structure and the name given to the structure. Further, if the stereochemistry of a structure or a portion of a structure is not indicated with, for example, bold or dashed lines, the structure or portion of the structure is to be interpreted as encompassing all stereoisomers of it.

Table I describes exemplary compounds according to formula I.

TABLE I

Synthesis of

General procedure

Scheme 1.

As shown in scheme 1 above, Q can be H, halogen, hydroxy, cyano, or Q'; q' may be lower alkyl, lower alkenyl, lower alkynyl, cycloalkyl, phenyl, cycloalkenyl, heterocycloalkyl or heteroaryl, optionally substituted with one or more Q^aSubstitution; q^aMay be Q^bOr Q^c；Q^bCan be halogen, oxo, hydroxy, -CN, -SCH₃、–S(O)₂CH₃or-S (= O) CH₃；Q^cMay be Q^dOr Q^e(ii) a Or two Q^aTogether form a bicyclic ring system, optionally substituted with one or more Q^bOr Q^cSubstitution; q^dMay be-O (Q)^e)、–S(=O)₂(Q^e)、–C(=O)N(Q^e)₂、–S(O)₂(Q^e)、–C(=O)(Q^e)、–C(=O)O(Q^e)、–N(Q^e)₂；–N(Q^e)C(=O)(Q^e)、–N(Q^e)C(=O)O(Q^e) or-N (Q)^e)C(=O)N(Q^e)₂；Q^eMay each independently be H or Q^e’；Q^e' may each independently be lower alkyl, phenyl, benzyl, lower haloalkyl, lower alkoxy, cycloalkyl, cycloalkenyl, heterocycloalkyl, or heteroaryl, optionally substituted with one or more Q^fSubstitution; q^fMay be Q^gOr Q^h；Q^gCan be halogen, hydroxy, cyano, oxo or-C (= O) (Q)^h)；Q^hCan beLower alkyl, lower haloalkyl, lower alkoxy, amino, phenyl, benzyl, cycloalkyl, heterocycloalkyl, or heteroaryl, optionally substituted with one or more QⁱSubstitution; and QⁱMay be halogen, hydroxy, cyano, lower alkyl, lower haloalkyl or lower alkoxy.

As shown in scheme 1 above, Y can be C (R)¹)₂(C(R¹’)₂)_m(ii) a m may be 0 or 1; r¹May each be H or R^1a；R^1aMay each independently be lower alkyl, lower alkoxy, phenyl, benzyl, heteroaryl, cycloalkyl, heterocycloalkyl or cycloalkyl lower alkyl, optionally substituted with one or more R^1a' substitution; r^1a' may be halogen, lower alkyl, lower haloalkyl, lower alkoxy, lower hydroxyalkyl, oxo, hydroxy or cyano; r¹' may each independently be H, lower alkyl or lower haloalkyl; r²May independently be H or R^2a；R^2aMay independently be lower alkyl, lower haloalkyl, lower alkoxy, lower hydroxyalkyl, cyano lower alkyl, cycloalkyl or heterocycloalkyl; or R^2aAnd R^1aTogether form a ring, which is optionally substituted with one or more of the following groups: halogen, lower alkyl, cyano lower alkyl, hydroxy, lower haloalkyl, lower hydroxyalkyl, lower alkoxy, lower alkylamino or lower dialkylamino; r³May independently be H or R^3a；R^3aMay independently be lower alkyl, lower haloalkyl, lower alkoxy, lower hydroxyalkyl, cyano lower alkyl, C (= O) R^3a' or S (= O)₂R^3a'; and R is^3a' may each independently be H or lower alkyl.

As shown in scheme 1 above, to 2-halo-5H-pyrrolo [2,3-b ]]Pyrazines in 1, 4-bisAdding 2.0M NaOH aqueous solution and formaldehyde aqueous solution into partial suspension of the alkaneAnd (4) liquid. The reaction mixture was stirred at room temperature overnight, the organics were evaporated under reduced pressure, the aqueous layer was neutralized with 1.0M HCl and extracted with EtOAc (2 ×). The combined organics were then concentrated and precipitated from the aqueous layer, collected by filtration and dried.

To a suspension of the product from the above reaction in THF (150mL) was added aqueous NaOH, stirred overnight, and the organics removed under reduced pressure as shown in scheme 1 above. The aqueous residue is then brought to pH4 with 1.0M aqueous HCl, the precipitate is collected by filtration and washed with H₂And (4) flushing.

By concentrating H as shown in scheme 1 above₂SO₄Adding CrO₃Then with H₂O dilution to prepare jones reagent stock. To a partial suspension of the product obtained from the above reaction in acetone was then added jones reagent. The reaction mixture was stirred, quenched with i-PrOH, filtered through celite, and rinsed with acetone. The filtrate was concentrated to give the aldehyde, which was used without further purification. To a solution of the aldehyde in cooled DMF was added NaH, the reaction mixture was stirred at room temperature, cooled, and 2- (trimethylsilyl) ethoxymethyl chloride (SEMCl) was added slowly. The reaction mixture was then warmed to room temperature, stirred, and then washed with H₂O quenched and extracted with EtOAc. The combined organics were then washed with H₂O and brine, MgSO₄Dried, concentrated and the residue of the aldehyde purified by chromatography.

As shown in scheme 1 above, a mixture of the aldehyde and substituted boronic acid, tricyclohexylphosphine, palladium (II) acetate, and tripotassium phosphate was heated overnight, cooled, filtered, washed with EtOAc, and concentrated under reduced pressure. The residue was purified by silica gel chromatography.

To the aldehyde in the cooled 1, 4-bis as shown in scheme 1 aboveTo the alkane/water solution was added sulfamic acid followed by dropwise addition of an aqueous solution of sodium chlorite and potassium dihydrogen phosphate. After the addition is finished, the reaction is carried outThe mixture was warmed to room temperature, stirred, and partitioned between water and ethyl acetate. The organic layer was washed with brine, dried over sodium sulfate, and concentrated under reduced pressure. The residue was triturated with hexane to isolate the carboxylic acid.

To the carboxylic acid in CH as shown in scheme 1 above₂Cl₂To the solution of (a) was added EDC, 4- (dimethylamino) pyridine and amine. The reaction mixture was stirred at room temperature overnight with H₂Diluting with O and using CH₂Cl₂And (4) extracting. The combined organics were washed with brine, washed with Na₂SO₄Dried and concentrated. The residue was purified by chromatography to give the condensed ester product.

As shown in scheme 1 above, to the ester is added THF/H₂Adding LiOH & H into O solution₂O, and subsequently acidifying the reaction mixture with aqueous acid and with CH₂Cl₂And (4) extracting. The combined organics were washed with Na₂SO₄Drying, concentration and chromatography of the residue afforded the acid product.

To the acid in CH as shown in scheme 1 above₂Cl₂To the solution of (a) was added EDC, 4- (dimethylamino) pyridine and amine. The reaction mixture was stirred at room temperature and H was added₂Diluting with O and CH₂Cl₂And (4) extracting. The combined organics were washed with brine, washed with Na₂SO₄Dried and concentrated. The residue was purified by chromatography to give the bisamide product.

As shown in scheme 1 above, to bisamides at CH₂Cl₂Trifluoroacetic acid was added to the solution to remove the SEM protecting groups, followed by stirring at room temperature and concentration. The residue can be dissolved in MeOH/H₂O, and Et is added₃And N is added. The reaction mixture was stirred at room temperature overnight, then concentrated, and the residue was purified by chromatography to give the bisamide.

Detailed description the specific methods for the synthesis of pyrrolo [2,3-b ] pyrazin-5-yl) starting materials are as follows:

step 1

To 2-bromo-5H-pyrrolo [2,3-b ]]Pyrazine (5.0g,25.2mmol) in 1, 4-bisTo a partial suspension of an alkane (100mL) was added 2.0M aqueous NaOH (25mL,50.0mmol) and 37% aqueous formaldehyde (19mL,252 mmol). The dark homogeneous reaction mixture was stirred at room temperature overnight. The organics were evaporated under reduced pressure. The aqueous layer was neutralized with 1.0M HCl and extracted with EtOAc (2 ×). The combined organics were concentrated to give 2.6g of an orange solid. After standing, a thick brown precipitate formed in the aqueous layer. The precipitate was collected by filtration and dried. The brown solid was extracted with hot 10% MeOH/EtOAC (3X 200 mL). The extracts were combined and evaporated to give a further 3.05g of an orange solid. The total yield was 5.65g (87%) of (2-bromo-7-hydroxymethyl-pyrrolo [2, 3-b)]Pyrazin-5-yl) -methanol.

Step 2

To (2-bromo-7-hydroxymethyl-pyrrolo [2, 3-b)]To a suspension of pyrazin-5-yl) -methanol (5.65g,21.9mmol) in THF (150mL) was added 2.0M aqueous NaOH (33mL,66 mmol). The homogeneous reaction mixture was stirred overnight, followed by removal of organics under reduced pressure. The aqueous residue was adjusted to pH4 with 1.0M aqueous HCl. The resulting precipitate was collected by filtration and washed with H₂O rinse to give 3.68g of a yellow solid. The filtrate was extracted with EtOAc (2 ×), and the organics were concentrated under reduced pressure to give a further 0.92g of yellow solid. The total yield was 4.60g (92%) (2-bromo-5H-pyrrolo [2, 3-b)]Pyrazin-7-yl) -methanol.

Step 3

By careful addition of concentrated H₂SO₄(2.3mL) CrO was added₃(2.67g), followed by H₂Stock solutions of jones reagent (2.67M) were prepared by diluting O to 10 mL. To (2-bromo-5H-pyrrolo [2, 3-b)]Pyrazin-7-yl) -methanol (4.6g,20.1mmol) in a portion of acetone (300mL) was added slowly to Jones' reagent (9mL,24.0 mmol). During the addition, the raw materials gradually dissolved and a thick green precipitate formed. The reaction mixture was stirred for 15 minutes, then quenched with i-PrOH (2mL) and filtered through celite, rinsing with acetone. The filtrate was concentrated to give 4.76g of 2-bromo-5H-pyrrolo [2,3-b ]]Pyrazine-7-carbaldehyde, a yellow-orange solid, which was used without further purification. To a solution of the solid in DMF (50mL) at 0 ℃ was added NaH (60% solution in mineral oil, 1.2g,30.1 mmol). The reaction mixture was stirred at room temperature for 30 minutes, then cooled to 0 ℃ and 2- (trimethylsilyl) ethoxymethyl chloride (4.3mL,24.1mmol) was added slowly. The reaction mixture was warmed to room temperature and stirred for 1 hour, then taken up with H₂O extraction and extraction with EtOAc (3 ×). The combined organic matter is treated with H₂O (3X) and brine, followed by MgSO₄Dried and concentrated. Passing the residue through SiO₂Chromatographically purify (20% -30% EtOAc/hexanes) to isolate 3.82g (53%) of 2-bromo-5- (2-trimethylsilyl-ethoxymethyl) -5H-pyrrolo [2,3-b ]]Pyrazine-7-carbaldehyde as a yellow solid.

Method 2

Step 1

2-bromo-5- (2-trimethylsilyl-ethoxymethyl) -5H-pyrrolo [2,3-b ] pyrazine-7-carbaldehyde (0.33g,0.93mmol), cyclopropylboronic acid (0.12g,1.39mmol), tricyclohexylphosphine (0.026g,0.09mmol), palladium (II) acetate (0.01g,0.046mmol), and tripotassium phosphate (0.63g,2.97mmol) were purged with argon in a mixture of 4mL of toluene and 0.5mL of water for 5 minutes, followed by heating at 100 ℃ for 18 hours. The cooled mixture was filtered through a celite column, washed with EtOAc, and concentrated under reduced pressure. The residue was purified by silica gel chromatography eluting with 10% EtOAc/hexanes to give 0.24g (81%) of 2-cyclopropyl-5- (2-trimethylsilyl-ethoxymethyl) -5H-pyrrolo [2,3-b ] pyrazine-7-carbaldehyde as a yellow powder.

Step 2

To 2-cyclopropyl-5- (2-trimethylsilyl-ethoxymethyl) -5H-pyrrolo [2,3-b ] at 0 ℃]Pyrazine-7-carbaldehyde (0.24g,0.75mmol) in 1, 4-bisTo a solution of alkane (10mL) and water (2mL) was added sulfamic acid (0.44g,4.54 mmol). A solution of sodium chlorite (0.09g,0.98mmol) and potassium dihydrogen phosphate (1.22g,9.0mmol) in 6mL of water was then added dropwise. After the addition was complete, the reaction mixture was warmed to room temperature and stirred for 2 hours, then partitioned between water and ethyl acetate. The organic layer was washed with brine, dried over sodium sulfate and concentrated under reduced pressure. The residue was triturated with hexane to give 0.22g (87%) of 2-cyclopropyl-5- (2-trimethylsilyl-ethoxymethyl) -5H-pyrrolo [2,3-b ]]Pyrazine-7-carboxylic acid as a pale yellow powder.

The following examples illustrate the preparation and biological evaluation of compounds within the scope of the present invention. The following examples and preparations are provided to enable those skilled in the art to more clearly understand and practice the present invention. They should not be construed as limiting the scope of the invention but merely as being illustrative and representative thereof.

Pharmaceutical compositions and administration

The compounds of the present invention may be formulated into a variety of oral dosage forms and carriers. The oral administration can be in the form of tablets, coated tablets, dragees, hard and soft gelatine capsules, solutions, emulsions, syrups or suspensions. The compounds of the present invention are effective when administered by other routes of administration, including continuous (intravenous drip) topical parenteral, intramuscular, intravenous, subcutaneous, transdermal (which may include a penetration enhancer), buccal, nasal, inhalation, and suppository administration, and the like. The preferred mode of administration is generally oral using a convenient daily dosing regimen which can be adjusted according to the degree of affliction and the patient's response to the active ingredient.

The compounds of the present invention and their pharmaceutically acceptable salts can be formulated into pharmaceutical compositions and unit dosages with 1 or more conventional excipients, carriers, or diluents. The pharmaceutical compositions and unit dosage forms may be comprised of conventional ingredients in conventional proportions, with or without additional active compounds or substances, and the unit dosage forms may contain any suitable effective amount of the active ingredient commensurate with the intended daily dosage range to be employed. The pharmaceutical compositions may be used as solids (e.g., tablets or filled capsules), semisolids, powders, sustained release formulations, or liquids (e.g., solutions, suspensions, emulsions, elixirs, or oral filled capsules); or in the form of suppositories for rectal or vaginal administration; or in the form of sterile injectable solutions for parenteral use. Typical formulations will contain from about 5% to about 95% active compound (w/w). The term "formulation" or "dosage form" includes both solid and liquid formulations of the active compound, and those skilled in the art will understand that the active ingredient can be present in different formulations depending on the target organ or tissue and depending on the desired dosage and pharmacokinetic parameters.

The term "excipient" as used herein refers to a compound used in the preparation of pharmaceutical compositions, which are generally safe, non-toxic and neither biologically nor otherwise undesirable, and which include veterinary and human pharmaceutically acceptable excipients. The compounds of the present invention can be administered alone, but will generally be administered in admixture with 1 or more suitable pharmaceutical excipients, diluents or carriers selected with regard to the intended route of administration and standard pharmaceutical practice.

By "pharmaceutically acceptable" it is meant that it is useful in the preparation of pharmaceutical compositions that are generally safe, non-toxic and neither biologically nor otherwise undesirable, and that include veterinary as well as human pharmaceutical uses.

The "pharmaceutically acceptable salt" form of the active ingredient may also initially confer desirable pharmacokinetic properties not found in the non-salt form of the active ingredient, and may even positively influence the pharmacodynamics of the active ingredient with respect to its therapeutic activity in vivo. The phrase "pharmaceutically acceptable salt" of a compound refers to a salt that is pharmaceutically acceptable and that possesses the desired pharmacological activity of the parent compound. The salt comprises: (1) formed with inorganic acids (e.g., hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, and the like) or with organic acids (e.g., acetic acid, propionic acid, hexanoic acid, cyclopentanepropionic acid, glycolic acid, pyruvic acid, lactic acid, malonic acid, succinic acid, malic acid, maleic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, 3- (4-hydroxybenzoyl) benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, 1, 2-ethane-disulfonic acid, 2-hydroxyethanesulfonic acid, benzenesulfonic acid, 4-chlorobenzenesulfonic acid, 2-naphthalenesulfonic acid, 4-toluenesulfonic acid, camphorsulfonic acid, 4-methylbicyclo [2.2.2] -oct-2-ene-1-carboxylic acid, glucoheptonic acid, 3-phenylpropionic acid, trimethylacetic acid, t-butylacetic acid, dodecylsulfuric acid, and the like), Gluconic acid, glutamic acid, hydroxynaphthoic acid, salicylic acid, stearic acid, muconic acid, etc.); or (2) salts formed when the acidic proton present in the parent compound is replaced by a metal ion (e.g., an alkali metal ion, alkaline earth ion, or aluminum ion) or is coordinated to an organic base (e.g., ethanolamine, diethanolamine, triethanolamine, tromethamine, N-methylglucamine, and the like).

Solid form preparations include powders, tablets, pills, capsules, cachets, suppositories, and dispersible granules. A solid carrier can be 1 or more substances which may also act as diluents, flavoring agents, solubilizers, lubricants, suspending agents, binders, preservatives, tablet disintegrating agents, or an encapsulating material. In powders, the carrier is usually a finely divided solid which is in admixture with the finely divided active ingredient. In tablets, the active ingredient is usually mixed with a carrier having the necessary binding capacity in suitable proportions and compacted in the shape and size desired. Suitable carriers include, without limitation, magnesium carbonate, magnesium stearate, talc, sugar, lactose, pectin, dextrin, starch, gelatin, tragacanth, methylcellulose, sodium carboxymethylcellulose, a low melting wax, cocoa butter, and the like. In addition to the active ingredients, solid form preparations may contain colorants, flavors, stabilizers, buffers, artificial and natural sweeteners, dispersants, thickeners, solubilizing agents, and the like.

Liquid preparations are also suitable for oral administration and include emulsions, syrups, elixirs, aqueous solutions, aqueous suspensions. These include solid form preparations which are intended to be converted, prior to use, to liquid form preparations. Emulsions may be prepared in solution, for example in aqueous propylene glycol solution, or they may contain emulsifying agents, for example lecithin, sorbitan monooleate or acacia. Aqueous solutions can be prepared by dissolving the active ingredient in water and adding suitable colorants, flavors, stabilizers, and thickening agents. Aqueous suspensions can be prepared by dispersing the finely divided active component in water with viscous material, such as natural or synthetic gums, resins, methylcellulose, sodium carboxymethylcellulose, and other known suspending agents.

The compounds of the invention may be formulated for parenteral administration (e.g., by injection, e.g., bolus injection or continuous infusion) and they may be in unit dosage form in ampoules, pre-filled syringes, small volume infusion or in multi-dose containers with an added preservative. The compositions may take such forms as suspensions, solutions, or emulsions in oily or aqueous vehicles, for example, as solutions in aqueous polyethylene glycol. Examples of oily or nonaqueous carriers, diluents, solvents or vehicles include propylene glycol, polyethylene glycol, vegetable oils (e.g., olive oil), and injectable organic esters (e.g., ethyl oleate), and which may contain formulatory agents such as preservatives, wetting, emulsifying or suspending, stabilizing and/or dispersing agents. Alternatively, the active ingredient may be in powder form for constitution with a suitable vehicle, e.g., sterile, pyrogen-free water, before use, by aseptic isolation of sterile solid or by lyophilization from solution.

The compounds of the invention may be formulated for topical administration to the epidermis as ointments, creams or lotions, or as a transdermal patch. For example, ointments and creams may be formulated with an aqueous or oily base to which suitable thickening and/or gelling agents are added. Lotions may be formulated with an aqueous or oily base and will in general also contain 1 or more emulsifying agents, stabilizing agents, dispersing agents, suspending agents, thickening agents, or coloring agents. Formulations suitable for topical administration in the mouth include: lozenges comprising the active agent in a flavored base (usually sucrose and acacia or tragacanth); pastilles comprising the active ingredient in an inert base (e.g. gelatin and glycerin or sucrose and acacia); and mouthwashes comprising the active ingredient in a suitable liquid carrier.

The compounds of the present invention may be formulated for administration as suppositories. The low melting wax (e.g., a mixture of fatty acid glycerides and cocoa butter) is first melted and the active ingredient is dispersed uniformly, such as by stirring. The molten homogeneous mixture is then poured into a mold of conventional size, allowed to cool and solidify.

The compounds of the invention may be formulated for vaginal administration. Pessaries, tampons, creams, gels, pastes, foams or sprays containing such carriers in addition to the active ingredient are known in the art to be suitable.

The compounds of the present invention may be formulated for nasal administration. Solutions and suspensions may be applied directly to the nasal cavity by conventional means, for example with a dropper, pipette or nebulizer. The formulations may be provided in single or multiple dose forms. In the latter case of a dropper or pipette, this may be achieved by the patient administering a suitable, predetermined volume of solution or suspension. In the case of a nebulizer, this can be achieved, for example, by a metered nebulization spray pump.

The compounds of the invention may be formulated for aerosol administration, particularly to the respiratory tract, and include intranasal administration. The compounds typically have a small particle size, for example, on the order of 5 microns or less. The particle size may be obtained by methods known in the art, for example by micronisation. The active ingredient is provided in pressurized packs with a suitable propellant, e.g., a chlorofluorocarbon (CFC), e.g., dichlorodifluoromethane, trichlorofluoromethane, or dichlorotetrafluoroethane, or carbon dioxide or other suitable gas. The aerosol may also suitably contain a surfactant, for example lecithin. The dosage of the drug can be controlled by a metering valve. Alternatively, the active ingredient may be provided in the form of a dry powder, for example a powder mix of the compound in a suitable powder base such as lactose, starch derivatives such as hydroxypropylmethyl cellulose and polyvinylpyrrolidone (PVP). The powder carrier will form a gel in the nasal cavity. The powder compositions may be in unit dosage form, for example in capsules or cartridges such as gelatin or blister packs, from which the powder may be administered by means of an inhaler.

When desired, the formulations can be prepared with enteric coatings suitable for sustained or controlled release administration of the active ingredient. For example, the compounds of the present invention can be formulated into transdermal or subcutaneous drug delivery devices. These delivery systems are advantageous when sustained release of the compound is necessary and when patient compliance with the treatment regimen is critical. The compounds in transdermal delivery systems are often attached to a solid support that adheres to the skin. The compound of interest can also be combined with a penetration enhancer such as azone (1-dodecylaza-cycloheptan-2-one). The sustained release drug delivery system is inserted subcutaneously into the subcutaneous layer by surgery or injection. Subcutaneous implants encapsulate compounds in a lipid soluble film (e.g., silicone rubber) or a biodegradable polymer (e.g., polylactic acid).

Suitable formulations with pharmaceutical carriers, diluents and excipients are described in Remington, The Science and Practice of Pharmacy 1995, mack publishing Company, 19 th edition, Easton, Pennsylvania, edited by e.w. The skilled formulation scientist, given the teachings of the specification, will vary the formulation to provide a wide variety of formulations for a particular route of administration without destabilizing or otherwise compromising the therapeutic activity of the compositions of the present invention.

For example, it is within the ordinary skill in the art that the compounds of the present invention can be readily modified to be more stable in water or other vehicles by minor modifications (salt formation, esterification, etc.). It is also within the ordinary skill in the art to vary the route of administration and dosage regimen of a particular compound in order to control the pharmacokinetics of the compounds of the present invention and thereby maximize the beneficial effects in the patient.

The term "therapeutically effective amount" as used herein refers to the amount required to reduce the symptoms of a disease in an individual. The dosage is adjusted according to the individual requirements in each particular case. The dosage can vary within wide limits depending on a number of factors, such as the severity of the disease being treated, the age and general health of the patient, other drugs being used while the patient is being treated, the route and form of administration, and the preferences and experience of the attending physician. For oral administration, daily dosages between about 0.01 to about 1000mg/kg body weight per day should be suitable for monotherapy and/or combination therapy. A preferred daily dosage is between about 0.1 and about 500mg/kg body weight, more preferably between about 0.1 and about 100mg/kg body weight per day, and most preferably between about 1.0 and about 10mg/kg body weight per day. Thus, for administration to a 70kg human, the dosage range will be about 7mg to 0.7g per day. The daily dose can be administered as a single dose or in divided doses, usually between 1 and 5 administrations per day. Typically, treatment is initiated at smaller doses than the optimal dose of the compound. Thereafter, the dose is increased in small increments until the optimum effect is achieved for each patient. The ordinarily skilled artisan in treating the diseases described herein will be able to determine, without undue experimentation and relying on his or her personal knowledge, experience, and content of this application, a therapeutically effective amount of a compound of the invention for a given disease and patient.

The pharmaceutical formulation is preferably in unit dosage form. In this form, the preparation is subdivided into unit doses containing appropriate amounts of the active ingredient. The unit dosage form can be a packaged preparation, the package containing a discrete number of the preparation, for example, tablets, capsules, and powders in vials or ampoules. The unit dosage form can also be a capsule, tablet, cachet, or lozenge itself, or it can be the appropriate number of any of these in packaged form.

Indications and treatment methods

Examples

Abbreviations

Common abbreviations include: acetyl (Ac), azo-bis-isobutyronitrile (AIBN), atmospheric pressure (Atm), 9-borabicyclo [3.3.1]Nonane (9-BBN or BBN), tert-butoxycarbonyl (Boc), di-tert-butyl dicarbonate or Boc anhydride (Boc)₂O), benzyl (Bn), butyl (Bu), chemical Abstract registry number (CASRN), benzyloxycarbonyl (CBZ or Z), Carbonyldiimidazole (CDI), 1, 4-diazabicyclo [2.2.2]Octane (DABCO), diethylaminosulfur trifluoride (DAST), dibenzylidene acetone (dba), 1, 5-diazabicyclo [4.3.0]Non-5-ene (DBN), 1, 8-diazabicyclo [5.4.0]Undec-7-ene (DBU), N ' -Dicyclohexylcarbodiimide (DCC), 1, 2-Dichloroethane (DCE), Dichloromethane (DCM), diethyl azodicarboxylate (DEAD), diisopropyl azodicarboxylate (DIAD), diisobutylaluminum hydride (DIBAL or DIBAL-H), Diisopropylethylamine (DIPEA), N-Dimethylacetamide (DMA), 4-N, N-Dimethylaminopyridine (DMAP), N-Dimethylformamide (DMF), dimethyl sulfoxide (DMSO), 1' -bis- (diphenylphosphino) ethane (dppe), 1' -bis- (diphenylphosphino) ferrocene (dppf), 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride (EDCI), Ethyl (Et), ethyl acetate (EtOAc), ethanol (EtOH), 2-ethoxy-2H-quinoline-1-carboxylic acid Ethyl Ester (EEDQ), diethyl ether (Et)₂O), O- (7-azabenzotriazol-1-yl) -N, N, N 'N' -tetramethylureaHexafluorophosphate acetic acid (HATU), acetic acid (HOAc), 1-N-hydroxybenzotriazole (HOBt), High Pressure Liquid Chromatography (HPLC), Isopropanol (IPA), lithium hexamethyldisilazane (LiHMDS), methanol (MeOH), melting point (mp), MeSO₂- (methanesulfonyl or Ms), methyl (Me), acetonitrile (MeCN), m-chloroperbenzoic acid (MCPBA), mass spectrometry (Ms or MS), methyl tert-butyl ether (MTBE), N-bromosuccinimide (NBS), N-carboxy anhydride (NCA), N-chlorosuccinimide (NCS), N-methylmorpholine (NMM), N-methylpyrrolidone (NMP), pyridinium chlorochromate (PCC), Pyridinium Dichromate (PDC), phenyl (Ph), propyl (Pr), isopropyl (i-Pr), pounds per square inch (psi), pyridine (pyr), room temperature (RT or RT), 2-trimethylsilanyl-ethoxymethyl (SEM), 2- (trimethylsilyl) ethoxymethyl chloride (SEMCl), t-butyldimethylsilyl or t-BuMe₂Si (TBDMS), triethylamine (TEA or Et)₃N), 2,6, 6-tetramethylpiperidine 1-oxyl (TEMPO), trifluoromethanesulfonic or CF₃SO₂- (Tf), trifluoroacetic acid (TFA), 1' -bis-2, 2,6, 6-tetramethylheptane-2, 6-dione (TMHD), O-benzotriazol-1-yl-N, N, N ', N ' -tetramethylureaTetrafluoroborate (TBTU), Thin Layer Chromatography (TLC), Tetrahydrofuran (THF), trimethylsilane or Me₃Si (TMS), p-toluenesulfonic acid monohydrate (TsOH or pTsOH), 4-Me-C₆H₄SO₂-or tosyl (Ts), N-urethane-N-carboxy anhydride (UNCA). The usual Nomenclature (including the prefixes positive (n), iso (i-), secondary (sec-), tertiary (tert-), and neo) has its usual meaning when used with alkyl moieties (j.rigaudy and d.p.klesney, Organic Chemistry Nomenclature (Nomenclature in Organic Chemistry), IUPAC 1979Pergamon Press, Oxford).

Preparation examples

Example 1

2-cyclopropyl-5H-pyrrolo [2,3-b ] pyrazine-7-carboxylic acid (2-ethylcarbamoyl-1-methyl-ethyl) -amide

Step 1

To 3- { [ 2-cyclopropyl-5- (2-trimethylsilyl-ethoxymethyl) -5H-pyrrolo [2,3-b]Pyrazine-7-carbonyl]-amino } -butyric acid (0.14g,0.33mmol) in CH₂Cl₂To a solution (5mL) were added EDC (0.083g,0.43mmol), 4- (dimethylamino) pyridine (0.053g,0.43mmol) and ethylamine hydrochloride (0.035g,0.43 mmol). The reaction mixture was stirred at room temperature overnight, then with H₂Diluting with O and using CH₂Cl₂And (4) extracting. The combined organics were washed with brine, washed with Na₂SO₄Dried and concentrated. Passing the residue through SiO₂Chromatographic purification (2% MeOH/CH)₂Cl₂) To yield 0.114g (78%) of 2-cyclopropyl-5- (2-trimethylsilyl-ethoxymethyl) -5H-pyrrolo [2, 3-b%]Pyrazine-7-carboxylic acid (2-ethylcarbamoyl-1-methyl-ethyl) -amide as an oil.

Step 2

To 2-cyclopropyl-5- (2-trimethylsilyl-ethoxymethyl) -5H-pyrrolo [2,3-b]Pyrazine-7-carboxylic acid (2-ethylcarbamoyl-1-methyl-ethyl) -amide (0.114g,0.25mmol) in CH₂Cl₂To the solution (8mL) was added trifluoroacetic acid (1.0 mL). The reaction mixture was stirred at room temperature overnight and then concentrated. The residue was dissolved in MeOH (7mL) and H₂O (0.5mL) and Et was added₃N (1 mL). The reaction mixture was stirred at room temperature overnight and then concentrated. Passing the residue through SiO₂Chromatographic purification (4% MeOH/CH)₂Cl₂) To yield 0.012g (16%) of 2-cyclopropyl-5H-pyrrolo [2,3-b ]]Pyrazine-7-carboxylic acid (2-ethylcarbamoyl-1-methyl-ethyl) -amide as a white solid. MS (M + H)⁺=316；mp=182.0-184.0。

Example 2

2-cyclopropyl-5H-pyrrolo [2,3-b ] pyrazine-7-carboxylic acid (1-methyl-2-propylcarbamoyl-ethyl) -amide

2-cyclopropyl-5H-pyrrolo [2,3-b ]]Pyrazine-7-carboxylic acid (1-methyl-2-propylcarbamoyl-ethyl) -amide. Prepared as described in the previous example, substituting n-propylamine for the ethylamine hydrochloride. MS (M + H)⁺=330；mp=198.0-200.0。

Example 3

2-cyclopropyl-5H-pyrrolo [2,3-b ] pyrazine-7-carboxylic acid [ 1-methyl-2- (2,2, 2-trifluoro-ethylcarbamoyl) -ethyl ] -amide

2-cyclopropyl-5H-pyrrolo [2,3-b ]]Pyrazine-7-carboxylic acid [ 1-methyl-2- (2,2, 2-trifluoro-ethylcarbamoyl) -ethyl]-an amide. Prepared as described in example 1, substituting 2,2, 2-trifluoroethylamine hydrochloride for the ethylamine hydrochloride. MS (M + H)⁺=370；mp=230.0-232.0。

Example 4

2-cyclopropyl-5H-pyrrolo [2,3-b ] pyrazine-7-carboxylic acid (2-cyclopropylcarbamoyl-1-methyl-ethyl) -amide

2-cyclopropyl-5H-pyrrolo [2,3-b ]]Pyrazine-7-carboxylic acid (2-cyclopropylcarbamoyl-1-methyl-ethyl) -amide. Prepared as described in example 1, substituting cyclopropylamine for the ethylamine hydrochloride. MS (M + H)⁺=328；mp=236.0-238.0。

Example 5

2-cyclopropyl-5H-pyrrolo [2,3-b ] pyrazine-7-carboxylic acid (2-carbamoyl-1-cyclopropyl-2-methyl-propyl) -amide

Step 1

To a solution of isobutyronitrile (0.30g,4.35mmol) in THF (8mL) at-78 ℃ was added LiHMDS (1.0M in THF, 4.8mL,4.8 mmol). The light yellow reaction mixture was stirred at-78 ℃ for 30 minutes, followed by slow addition of 2-methyl-propane-2-sulfinic acid 1-cyclopropyl-methylene- (E) -ylamide (0.50g,2.90mmol) [ prepared according to WO2008/147800]Solution in THF (2 mL). The reaction mixture was stirred at-78 ℃ for 2 hours, followed by saturated NH₄Aqueous Cl was quenched and warmed to room temperature. The mixture was diluted with water and extracted with EtOAc (2 ×). The combined organics were washed with MgSO₄Dried and concentrated to give 0.70g of 2-methylpropane-2-sulfinic acid (2-cyano-1-cyclopropyl-2, 2-dimethyl-ethyl) -amide as a viscous colorless oil.

Step 2

To a solution of 2-methylpropane-2-sulfinic acid (2-cyano-1-cyclopropyl-2, 2-dimethyl-ethyl) -amide (0.70g,2.90mmol) in MeOH (5mL) at room temperature was added 4.0M HCl in bisAlkane solution (1.5mL,6.0 mmol). The reaction mixture was stirred at room temperature for 15 minutes and then concentrated to give 0.45g (89%,2 steps) of 3-amino-3-cyclopropyl-2, 2-dimethyl-propionitrile hydrochloride as a white solid.

Step 3

In a flask 2-cyclopropyl-5- ((2- (trimethylsilyl) ethoxy) methyl) -5H-pyrrolo [2, 3-b)]Pyrazine-7-carboxylic acid (120mg,0.36mmol), 3-amino-3-cyclopropyl-2, 2-dimethyl-propionitrile hydrochloride (75mg,0.43mmol), HOBt (54mg,0.40mmol) and EDC (77mg,0.40mmol) were combined. DMF (2mL) was then added followed by diisopropylethylamine (0.16mL,0.90 mmol). The reaction mixture was stirred at room temperature overnight, then quenched with water and extracted with EtOAc (3 ×). The combined organics were washed with water (3 ×), then MgSO₄Dried and concentrated. Passing the residue through SiO₂Chromatographic purification (30% -50% EtOAc/hexanes) afforded 121mg (74%) of 2-cyclopropyl-5- (2-trimethylsilyl-ethoxymethyl) -5H-pyrrolo [2,3-b ]]Pyrazine-7-carboxylic acid (2-cyano-1-cyclopropyl-2, 2-dimethyl-ethyl) -amide as an off-white foam.

Step 4

To 2-cyclopropyl-5- (2-trimethylsilyl-ethoxymethyl) -5H-pyrrolo [2,3-b]Pyrazine-7-carboxylic acid (2-cyano-1-cyclopropyl-2, 2-dimethyl-ethyl) -amide (163mg,0.36mmol) in EtOH (9mL) and H₂O (1mL) solution was added [ PtH (PMe)₂OH)(PMe₂O)₂H](15mg,0.036 mmol). The reaction mixture was heated at reflux for 6 hours. Additional catalyst (10mg,0.023mmol) was added and heating continued overnight. The reaction mixture was cooled to room temperature and concentrated. Passing the residue through SiO₂Chromatographic purification (50% -100% EtOAc/hexanes) afforded 108mg (64%) of 2-cyclopropyl-5- (2-trimethylsilyl-ethoxymethyl) -5H-pyrrolo [2,3-b ]]Pyrazine-7-carboxylic acid (2-carbamoyl-1-cyclopropyl-2-methyl-propyl) -amide as a viscous colorless oil.

Step 5

To 2-cyclopropyl-5- (2-trimethylsilyl-ethoxymethyl) -5H-pyrrolo [2,3-b]Pyrazine-7-carboxylic acid (2-carbamoyl-1-cyclopropyl-2-methyl-propyl) -amide (108mg,0.23mmol) in CH₂Cl₂To the solution (4mL) was added TFA (1 mL). The yellow reaction mixture was stirred for 3 hours and then concentrated. The residue was dissolved in CH₂Cl₂(4mL) and ethylenediamine (0.5mL) was added. The reaction mixture was stirred for 1 hour and then concentrated. Passing the residue through SiO₂Purify by chromatography (50% -100% EtOAc/hexanes to 5% MeOH/EtOAc) then use Et₂Trituration with O gave 44mg (56%) of 2-cyclopropyl-5H-pyrrolo [2, 3-b%]Pyrazine-7-carboxylic acid (2-carbamoyl-1-cyclopropyl-2-methyl-propyl) -amide as a white solid. MS (M + H)⁺=342；mp=222.0-224.0。

Example 6

2-cyclopropyl-5H-pyrrolo [2,3-b ] pyrazine-7-carboxylic acid carbamoylmethyl-amides

Step 1

To 2-cyclopropyl-5- ((2- (trimethylsilyl) ethoxy) methyl) -5H-pyrrolo [2, 3-b)]Pyrazine-7-carboxylic acid (0.20g,0.59mmol) in CH₂Cl₂To a solution (5mL) were added EDC (0.14g,0.72mmol), 4- (dimethylamino) pyridine (0.088g,0.72mmol) and aminoacetonitrile (0.041g,0.72 mmol). The reaction mixture was stirred at room temperature for 5 hours, followed by CH₂Cl₂Diluting with H₂O and brine wash. Adding Na to the organic matter₂SO₄Dried and concentrated. Passing the residue through SiO₂Chromatography (25% EtOAc/hexanes) afforded 0.194g (88%) of 2-cyclopropyl-5- (2-trimethylsilyl-ethoxymethyl) -5H-pyrrolo [2,3-b ]]Pyrazine-7-carboxylic acid cyanomethyl-amide as a white solid.

Step 2

To 2-cyclopropyl-5- (2-trimethylsilyl-ethoxymethyl) -5H-pyrrolo [2,3-b]Pyrazine-7-carboxylic acid cyanomethyl-amide (0.19g,0.50mmol) in CH₂Cl₂To the solution (6mL) was added trifluoroacetic acid (1.0 mL). The reaction mixture was stirred at room temperature overnight and then concentrated. The residue was dissolved in MeOH (10mL) and H₂O (2mL) and Et was added₃N (2 mL). The reaction mixture was stirred at room temperature overnight and then concentrated. The residue was triturated with EtOH to give 0.093g (77%) of 2-cyclopropyl-5H-pyrrolo [2,3-b ]]Pyrazine-7-carboxylic acid carbamoylmethyl groupAmide as a pale yellow solid. MS (M + H)⁺=260；mp=215.0-220.0。

Example 7

2-cyclopropyl-5H-pyrrolo [2,3-b ] pyrazine-7-carboxylic acid ((R) -1-dimethylcarbamoyl-ethyl) -amide

Step 1

(R) -2- (tert-Butoxycarbonylamino) propionic acid (1.0g,5.3mmol) and O-benzotriazol-1-yl-N, N, N ', N' -tetramethylureaTetrafluoroborate (1.8g,5.6mmol) was dissolved in 13mL of dichloromethane. Dimethylamine hydrochloride (0.64g,7.8mmol) and N, N-diisopropylethylamine (3.6mL,20.8mmol) were added and the mixture was stirred at room temperature for 16 h. The reaction mixture was diluted with dichloromethane and water was added. The layers were separated and the aqueous layer was extracted again with dichloromethane. The combined organic layers were washed with sodium chloride solution, dried over sodium sulfate and concentrated. The residue was purified by silica gel chromatography (ethyl acetate/hexane) to give 1.16g (100%) of tert-butyl ((R) -1-dimethylcarbamoyl-ethyl) -carbamate.

Step 2

((R) -1-dimethylcarbamoyl-ethyl) -carbamic acid tert-butyl ester (1.1g,5.1mmol) was dissolved in 26mL of cooled 4M HClIn an alkane solution. After 1.5 hours, the reaction was evaporated to give (R) -2-amino-N, N-dimethyl-propionamide hydrochloride, which was used without further purification.

Step 3

2-cyclopropyl-5H-pyridinePyrrolo [2,3-b]Pyrazine-7-carboxylic acid ((R) -1-dimethylcarbamoyl-ethyl) -amide. Prepared as described in steps 3 and 5 of example 5, substituting (R) -2-amino-N, N-dimethyl-propionamide hydrochloride for 3-amino-3-cyclopropyl-2, 2-dimethyl-propionitrile hydrochloride. MS (M + H)⁺= 302; mp = 236.0-239.0; elemental analysis showed C59.79, H6.26, N23.24, found C59.79, H6.15, N23.05.

Example 8

2-cyclopropyl-5H-pyrrolo [2,3-b ] pyrazine-7-carboxylic acid ((R) -1-methyl-2-oxo-piperidin-3-yl) -amide

Step 1

To (R) -3-aminopiperidin-2-one (1.00g,8.76mmol) in CH at room temperature₂Cl₂To a solution (15mL) were added triethylamine (1.28mL,9.2mmol) and di-tert-butyl dicarboxylate (2.01g,9.2 mmol). The reaction mixture was stirred at the same temperature for 12 hours, then concentrated under vacuum. The crude residue was diluted with ether (50mL) and filtered through a celite cartridge. The filtrate was evaporated to dryness and diluted with SiO₂Chromatographic purification (50g, EtOAc 100%) afforded 1.65g (88%) of tert-butyl (R) -2-oxopiperidin-3-ylcarbamate as a colorless foam.

Step 2

To a solution of (R) -tert-butyl 2-oxopiperidin-3-ylcarbamate (642mg,3.00mmol) in N, N-dimethylformamide (3mL) was added sodium hydride (132mg,3.3mmol) at room temperature. The reaction mixture was stirred for 30 min, followed by the addition of methyl iodide (206 μ L,3.3mmol) and stirring continued for 1 h. The reaction mixture was poured into water (50mL) and extracted with EtOAc (100mL) using a continuous extraction apparatus overnight. The organic extracts were evaporated to dryness in vacuo and the crude residue was taken up in SiO₂Chromatographic purification (23g, CH)₂Cl₂/MeOH/NH₄OH,100:0:0 to 94:5.7:0.3) to give 310mg (45%) of tert-butyl (R) -1-methyl-2-oxopiperidin-3-ylcarbamate as a colorless viscous oil.

Step 3

A solution of (R) -tert-butyl 1-methyl-2-oxopiperidin-3-ylcarbamate (310mg,1.36mmol) in 2,2, 2-trifluoroethanol (4.95mL,67.9mmol) was heated at 150 ℃ for 3 hours with the aid of microwaves. The solvent was evaporated to dryness and the crude residue was taken up in SiO₂Chromatographic purification (11g, CH)₂Cl₂/MeOH/NH₄OH,100:0:0 to 94:5.7:0.3) to give 120mg (69%) of (R) -3-amino-1-methyl-piperidin-2-one as a pale yellow oil.

Step 4

To 2-cyclopropyl-5- ((2- (trimethylsilyl) ethoxy) methyl) -5H-pyrrolo [2,3-b ] at room temperature]To a solution of pyrazine-7-carboxylic acid (150mg,0.45mmol) and (R) -3-amino-1-methyl-piperidin-2-one (115mg,0.90mmol) in DMF (2.4mL) was added triethylamine (0.19mL,1.35mmol) and PyBOP (257mg,0.50 mmol). The reaction mixture was stirred overnight, then diluted with EtOAc (30mL) and washed with water (4 × 30 mL). The combined organic extracts were dried (Na)₂SO₄) And evaporated in vacuo. The crude residue is treated with SiO₂Column chromatography purification (11g, CH)₂Cl₂/MeOH/NH₄OH,100:0:0 to 94:5.7:0.3) to yield 149mg (74%) of 2-cyclopropyl-5- (2-trimethylsilylethoxymethyl) -5H-pyrrolo [2,3-b ]]Pyrazine-7-carboxylic acid ((R) -1-methyl-2-oxo-piperidin-3-yl) -amide as a colorless viscous oil.

Step 5

To 2-cyclopropyl-5- (2-trimethylsilylethoxymethyl) -5H-pyrrolo [2,3-b ] at room temperature]To a solution of pyrazine-7-carboxylic acid ((R) -1-methyl-2-oxo-piperidin-3-yl) -amide (145mg,0.33mmol) in acetonitrile (14.2mL) were added 18-crown-6 (86.4mg,0.33mmol) and cesium fluoride (497mg,3.27 mmol). The reaction mixture was heated to reflux temperature for 48 hours, then cooled to room temperature and filtered through a celite pad. The filtrate was evaporated in vacuo and the crude residue was taken up inEtOAc (25mL) and water (25mL) were partitioned. The organic extracts were dried (Na)₂SO₄) And evaporated in vacuo. Passing the crude residue through SiO₂Column chromatography purification (25g, CH)₂Cl₂/MeOH/NH₄OH,100:0:0 to 90:9.5:0.5) to yield 60mg (59%) of 2-cyclopropyl-5H-pyrrolo [2,3-b ]]Pyrazine-7-carboxylic acid ((R) -1-methyl-2-oxo-piperidin-3-yl) -amide as a white solid. MS (M + H)⁺=314。

Example 9

2-cyclopropyl-5H-pyrrolo [2,3-b ] pyrazine-7-carboxylic acid ((R) -2-oxo-piperidin-3-yl) -amide

2-cyclopropyl-5H-pyrrolo [2,3-b ]]Pyrazine-7-carboxylic acid ((R) -2-oxo-piperidin-3-yl) -amide. Prepared according to the procedure outlined in example 8 (omitting steps 1-3), substituting (R) -3-amino-1-methyl-piperidin-2-one with (R) -3-aminopiperidin-2-one in step 4. MS (M + H)⁺=300。

Example 10

2-cyclopropyl-5H-pyrrolo [2,3-b ] pyrazine-7-carboxylic acid [ (R) -1- (2-cyano-ethyl) -2-oxo-piperidin-3-yl ] -amide

2-cyclopropyl-5H-pyrrolo [2,3-b ]]Pyrazine-7-carboxylic acid [ (R) -1- (2-cyano-ethyl) -2-oxo-piperidin-3-yl]-an amide. The title compound was isolated as a by-product of the procedure of example 9, but it can also be prepared as outlined in example 8, substituting acrylonitrile for methyl iodide in step 2. MS (M + H)⁺=353。

Example 11

2-cyclopropyl-5H-pyrrolo [2,3-b ] pyrazine-7-carboxylic acid ((R) -1-dimethylcarbamoyl-3-methyl-butyl) -amide

Step 1

To a solution of Boc-D-leucine monohydrate (2.0g,8.0mmol), triethylamine (5.6mL,40.1mmol) and dimethylamine hydrochloride (1.31g,16.0mmol) in DMF (15mL) was added PyBOP (4.59g,8.82mmol) at room temperature. The reaction mixture was stirred overnight, then diluted with EtOAc (50mL) and washed with water (4 × 50 mL). The combined organic extracts were dried (Na)₂SO₄) And evaporated in vacuo. The crude residue is treated with SiO₂Chromatography purification (80g, hexanes/EtOAc, 1:1) afforded 1.19g (58%) of ((R) -1-dimethylcarbamoyl-3-methyl-butyl) -carbamic acid tert-butyl ester as a colorless viscous oil.

Step 2

2-cyclopropyl-5H-pyrrolo [2,3-b ]]Pyrazine-7-carboxylic acid ((R) -1-dimethylcarbamoyl-3-methyl-butyl) -amide. Prepared as described in example 8, steps 3-5, substituting ((R) -1-methyl-2-oxopiperidin-3-ylcarbamic acid tert-butyl ester with ((R) -1-dimethylcarbamoyl-3-methyl-butyl) -carbamic acid tert-butyl ester. MS (M + H)⁺=344。

Example 12

2-cyclopropyl-5H-pyrrolo [2,3-b ] pyrazine-7-carboxylic acid [ (R) -1- (cyanomethyl-methyl-carbamoyl) -ethyl ] -amide

2-cyclopropyl-5H-pyrrolo [2,3-b ]]Pyrazine-7-carboxylic acid [ (R) -1- (cyanomethyl-aminomethyl-carbamic acidAcyl) -ethyl]-an amide. Prepared as described in example 11, substituting Boc-D-alanine for Boc-D-leucine monohydrate and methylaminoacetonitrile hydrochloride for dimethylamine hydrochloride. MS (M + H)⁺=327。

Example 13

2-cyclopropyl-5H-pyrrolo [2,3-b ] pyrazine-7-carboxylic acid ((S) -1-dimethylcarbamoyl-ethyl) -amide

Step 1

To 2-cyclopropyl-5- ((2- (trimethylsilyl) ethoxy) methyl) -5H-pyrrolo [2,3-b ] at room temperature]To a solution of pyrazine-7-carboxylic acid (250mg,0.75mmol) in DMF (6mL) were added triethylamine (0.52mL,3.75mmol), (S) -2-aminopropionic acid tert-butyl ester hydrochloride (136mg,0.75mmol) and PyBOP (429mg,0.83 mmol). The reaction mixture was stirred overnight, then poured into EtOAc (50mL) and washed with water (4 × 30 mL). The organic extracts were combined and dried (Na)₂SO₄) And evaporated under reduced pressure. Passing the crude residue through SiO₂Column chromatography purification (12g, hexanes/EtOAc, 7:3 to 1:1) afforded 285mg (85%) of (S) -2- (2-cyclopropyl-5- ((2- (trimethylsilyl) ethoxy) methyl) -5H-pyrrolo [2,3-b ]]Pyrazine-7-carboxamido) propionic acid tert-butyl ester as a viscous oil.

Step 2

A solution of tert-butyl (S) -2- (2-cyclopropyl-5- ((2- (trimethylsilyl) ethoxy) methyl) -5H-pyrrolo [2,3-b ] pyrazine-7-carboxamido) propionate (265mg,0.58mmol) in 2,2, 2-trifluoroethanol (4mL) was heated at 150 ℃ for 3 hours under microwave-assisted conditions. The solvent was removed in vacuo to give 230mg (98%) of (S) -2- (2-cyclopropyl-5- ((2- (trimethylsilyl) ethoxy) methyl) -5H-pyrrolo [2,3-b ] pyrazine-7-carboxamido) propionic acid as an oil.

Step 3

To (S) -2- (2-cyclopropyl-5- ((2- (trimethylsilyl) ethoxy) methyl) -5H-pyrrolo [2,3-b ] at room temperature]To a solution of pyrazine-7-carboxamido) propionic acid (253mg,0.63mmol), triethylamine (0.44mL,3.13mmol) and dimethylamine hydrochloride (102mg,1.25mmol) in DMF (5mL) was added PyBOP (358mg,0.69 mmol). The reaction mixture was stirred overnight, then diluted with EtOAc (50mL) and washed with water (4 × 25 mL). The organic extracts were combined and dried (Na)₂SO₄) Followed by evaporation in vacuo. Passing the crude residue through SiO₂Chromatography purification (24g, hexanes: EtOAc,3:7) afforded 190mg (70%) (S) -2-cyclopropyl-N- (1- (dimethylamino) -1-oxoprop-2-yl) -5- ((2- (trimethyl-silyl) ethoxy) methyl) -5H-pyrrolo [2,3-b]Pyrazine-7-carboxamide as a colorless viscous oil.

Step 4

To (S) -2-cyclopropyl-N- (1- (dimethylamino) -1-oxoprop-2-yl) -5- ((2- (trimethylsilyl) ethoxy) methyl) -5H-pyrrolo [2,3-b ] at room temperature]To a solution of pyrazine-7-carboxamide (150mg,0.35mmol) in acetonitrile (15.8mL) was added 18-crown-6 (92mg,0.35mmol) and cesium fluoride (528mg,3.48 mmol). The reaction mixture was heated to reflux temperature for 48 hours, then cooled to room temperature and filtered through a celite pad. The filtrate was evaporated in vacuo and the crude residue partitioned between EtOAc (25mL) and water (25 mL). The organic extracts were dried (Na)₂SO₄) And evaporated in vacuo. The crude residue was purified by preparative TLC (CH)₂Cl₂/MeOH/NH₄OH,90:9.5:0.5) to yield 48mg (48%) of 2-cyclopropyl-5H-pyrrolo [2,3-b ]]Pyrazine-7-carboxylic acid ((S) -1-dimethylcarbamoyl-ethyl) -amide as an off-white solid. MS (M + H)⁺=302。

Biological examples

JAK assay information

IC for Janus kinase (JAK) inhibition₅₀The determination of (1):

the enzymes and peptide substrates used were as follows:

JAK 1: recombinant human kinase domain from Invitrogen (Cat # PV4774)

JAK 3: recombinant human kinase domain from or prepared from Millipore (Cat #14-629) (810-

JAK 2: recombinant human kinase domain from Millipore (Cat #14-640)

Substrate: an N-terminally biotinylated 14-amino acid peptide from the activation loop of JAK1 having the peptide substrate sequence: Biotin-KAIETDKEYYTVKD

The test conditions used were as follows:

assay buffer: JAK kinase buffer: 50mM Hepes [ pH 7.2]，10mM MgCl₂1mM DTT, 1mg/ml BSA. The assay was performed in this buffer.

Test arrangement: kinase activity of all 3 JAK kinases was determined using radioactive end-point analysis and trace amounts³³P-ATP was measured. The assay was performed in 96-well polypropylene plates.

The experimental method comprises the following steps:

all concentrations were final concentrations in the reaction mixture and all incubations were performed at room temperature. The test procedure is as follows:

compounds were serially diluted in 100% DMSO, typically at 10x starting concentration of 1 mM. The final concentration of DMSO in the reaction was 10%.

The compounds were preincubated with enzyme (0.5nM JAK3 (commercially available), 0.2nM JAK3 (prepared), 1nM JAK2,5nM JAK1) for 10 min.

The reaction was initiated by adding a mixture of 2 substrates (ATP and peptide pre-mixed in JAK kinase buffer). In the JAK2/JAK3 assay, ATP and peptide were used at concentrations of 1.5uM and 50uM, respectively. The JAK1 assay was performed at an ATP concentration of 10uM and a peptide concentration of 50 uM.

The duration of the assay for JAK2 and JAK3 was 20 minutes. JAK1 assay was performed for 40 minutes. The reaction was stopped by adding 0.5M EDTA to a final concentration of 100mM using all 3 enzymes.

25ul of terminated reaction was transferred to 150ul in MgCl free, 96-well, 1.2um MultiScreen-BV filter plates₂-and CaCl₂A slurry of streptavidin-coated agarose beads at 7.5% (v/v) in 1x phosphate buffered saline containing 50mM EDTA.

After 30 minutes incubation, the beads were washed under vacuum with the following buffer:

wash 3-4 times with 200ul 2M NaCl.

Washing 3-4 times with 200ul 2M NaCl +1% (v/v) phosphoric acid.

Washed 1 time with water.

The washed plate was dried in an oven at 60 ℃ for 1-2 hours.

To each well of the filter plate, 70ul of Microscint 20 scintillation fluid was added and after at least 30 minutes of incubation, the radioactivity counts were measured in a Perkinelmer microplate scintillation counter.

Representative IC₅₀The results are in table II below:

table II.

Compound (I)	Ic50h-jak3(810 & 1124) -sf9-c without additives
		I-1	0.28694
I-2	0.19986
		I-3	0.90865
I-4	0.83647
		I-5	0.98581
I-6	0.11159
		I-7	0.06801
I-8	0.03966
		I-9	1.23713
I-10	0.09504
		I-11	0.31495
I-12	0.17802
		I-13	0.13796

SYK test information

Determination of IC for spleen tyrosine kinase (SYK) inhibition₅₀

The SYK kinase assay is a standard kinase assay adapted to the 96-well plate format. The experiment was performed in 96-well format for IC with 8 samples₅₀The samples represent 10 semilog dilutions and a 40 μ L reaction volume. This assay measures incorporation into N-terminally biotinylated peptide substrates³³P γ ATP, the biotinylated peptide substrate derived from a naturally occurring phosphorylated receptor consensus sequence (biotin-11 aa DY @). After stopping the reaction with EDTA and adding streptavidin-coated beads, the phosphorylated product was determined.

Test plate 96-well MultiScreen 0.65um Filter plate (Millipore Cat: MADVNOB10)

Streptavidin-coated beads streptavidin Sepharose TM, suspension 5.0mL, diluted in 50mM EDTA/PBS (1:100), (Amersham, Cat: 17-5113-01)

10mM of compound in 100% dimethyl sulfoxide (DMSO) at a final concentration of 0.003-100uM of compound in 10% DMSO

SYK RPA purified, truncated spleen tyrosine kinase aa 360-635 construct, stock solution 1mg/mL, MW 31.2KDa, final concentration: 0.0005. mu.M.

Peptide 1 biotinylated peptide was derived from the naturally occurring phosphorylated receptor consensus sequence (biotin-EPEGDYEEVLE), from a special order for QCB, 20mM stock solution, final concentration: 5.0. mu.M.

ATP adenosine-5' -triphosphate 20mM, (ROCHE catalog No. 93202720), final concentration 20. mu.M

Buffer HEPES 2-hydroxyethylpiperazine-2-ethanesulfonic acid (Sigma, cat # H-3375) final concentration 50mM HEPES pH7.5

BSA bovine serum albumin fraction V, no fatty acids (Roche Diagnostics GmbH, Cat. No. 9100221), diluted to a final concentration of 0.1%

EDTA storage 500mM, (GIBCO, catalog number 15575-038) final concentration 0.1mM

DTT 1, 4-dimercaptothreitol (Roche Diagnostics GmbH, Cat. No.: 197777) at a final concentration of 1mM

MgCl₂x 6H₂O MERCK, cat # 105833.1000, final concentration 10mM

Assay Dilution Buffer (ADB) 50mM HEPES,0.1mM EGTA,0.1mM sodium vanadate, 0.1mM beta-glycerophosphate, 10mM MgCl₂,1mM DTT,0,1%BSA,pH 7.5

Bead wash buffer 10g/L PBS (phosphate buffered saline) containing 2M NaCl +1% phosphoric acid.

The test method comprises the following steps:

26 μ L of ADB diluted purified recombinant human SYK360-635[0.5nM ] were mixed in 40 μ L volumes with 4 μ L of test compound [ usually 100 μ M-0.003 μ M ] at 10X concentration in [10% ] DMSO and the mixture incubated at room temperature for 10 min.

The kinase reaction was initiated by adding 10. mu.L of a 4 Xsubstrate mixture containing the DYE peptide substrate [0 or 5. mu.M ]]、ATP[20μM]And³³PγATP[2μCi/rxn]. After 15 min incubation at 30 ℃, the reaction was stopped by transferring 25 μ Ι _ of reaction sample to a 96-well 0.65 μ η millipore madpnob membrane/plate containing 200 μ Ι _ of 5mM EDTA and 20% streptavidin-coated beads in PBS.

Unbound radionuclide was removed under vacuum with 3 × 250 μ L2M NaCl; 2 × 250 μ L2 m nacl +1% phosphoric acid; 1X 250. mu. L H₂And O washing. After the last wash, the membrane/plate was transferred to a conversion plate, heat dried at 60 ℃ for 15 minutes, 50 μ L of scintillation cocktail was added to each well, and the amount of radioactivity was counted in a microplate reader after 4 hours.

Percent inhibition was calculated from the uninhibited enzyme ratio:

% inhibition =100/(1+ (IC)₅₀Inhibitor concentration)ⁿ)

IC was calculated using XLFit software (ID Business Solution Ltd., Guilford, Surrey, UK) with nonlinear curve fitting₅₀The value is obtained.

The foregoing invention has been described in some detail by way of illustration and example for purposes of clarity and understanding. It will be apparent that those skilled in the art can make changes and modifications within the scope of the appended claims. Accordingly, it is to be understood that the above description is intended to be illustrative, and not restrictive. The scope of the invention should, therefore, be determined not with reference to the above description, but instead should be determined with reference to the following appended claims, along with the full scope of equivalents to which such claims are entitled.

All patents, patent applications, and publications cited in this application are incorporated herein by reference in their entirety for all purposes to the same extent as if each individual patent, patent application, or publication were individually indicated to be incorporated by reference.

Claims

1. A compound of formula I or a pharmaceutically acceptable salt thereof,

wherein:

y is C (R)¹)₂(C(R¹’)₂)_m

m is 0 or 1;

R¹each is H or R^1a；

R¹' are each independently H, lower alkyl or lower haloalkyl;

R²independently is H or R^2a；

R³independently is H or R^3a；

R^3a' are each independently H or lower alkyl;

q is H, halogen, hydroxy, cyano or Q';

Q^ais Q^bOr Q^c；

Q^bIs halogen, oxo, hydroxy, -CN, -SCH₃、–S(O)₂CH₃or-S (= O) CH₃；

Q^cIs Q^dOr Q^e；

Q^eEach independently is H or Q^e’；

Q^fis Q^gOr Q^h；

Q^gIs halogen, hydroxy, cyano, oxo or-C (= O) (Q)^h)；

QⁱIs halogen, hydroxy, cyano, lower alkyl, lower haloalkyl or lower alkoxy.

2. The compound of claim 1, wherein Q is cycloalkyl or heterocycloalkyl, optionally substituted with one or more Q^aAnd (4) substitution.

3. The compound of claim 1 or 2, wherein Q is cyclopropyl, optionally substituted with one or more Q^aAnd (4) substitution.

4. The compound of any one of claims 1-3, wherein Q is cycloalkyl.

5. The compound of any one of claims 1-4, wherein one R¹Is lower alkyl, and the other R¹Is H.

6. The compound of any one of claims 1-5, wherein one R¹Is H, lower alkyl or cycloalkyl, or R^2aAnd R¹Together form a piperidinyl ring, and the other R¹Is H.

7. The compound of any one of claims 1-6, wherein m is 0.

8. The compound of any one of claims 1-4, wherein R¹Each independently is H, lower alkyl or cycloalkyl.

9. The compound of any one of claims 1-8, wherein R¹Is methyl, cyclopropyl or sec-butyl.

10. The compound of any one of claims 1-4, wherein R^2aAnd R^1aTogether form a ring, which is optionally substituted with lower alkyl, cyano or cyano lower alkyl.

11. The compound of any one of claims 1-4, wherein m is 1 and R is¹' are each H.

12. The compound of any one of claims 1-11Wherein R is²And R³Independently is H, lower alkyl, cycloalkyl, cyano lower alkyl or lower haloalkyl.

13. A compound selected from the following:

14. A compound selected from the following:

15. A method of treating an inflammatory or autoimmune disease comprising administering to a patient in need thereof a therapeutically effective amount of a compound of any one of claims 1-14.

16. The method of claim 15, further comprising administering an additional therapeutic agent selected from a chemotherapeutic or anti-proliferative agent, an anti-inflammatory agent, an immunomodulatory or immunosuppressive agent, a neurotrophic factor, an agent for treating cardiovascular disease, an agent for treating diabetes, or an agent for treating immunodeficiency disorders.

17. A method of treating rheumatoid arthritis comprising administering to a patient in need thereof a therapeutically effective amount of a compound of any one of claims 1-14.

18. A method of treating asthma comprising administering to a patient in need thereof a therapeutically effective amount of a compound of any one of claims 1-14.

19. A method of treating an immune disease comprising administering to a patient in need thereof a therapeutically effective amount of a compound of any one of claims 1-14, the immune disease comprising lupus, multiple sclerosis, rheumatoid arthritis, psoriasis, type I diabetes, organ transplant complications, xenotransplantation, diabetes, cancer, asthma, atopic dermatitis, autoimmune thyroid disease, ulcerative colitis, crohn's disease, alzheimer's disease, and leukemia.

20. A pharmaceutical composition comprising a compound according to any one of claims 1-14 in admixture with at least 1 pharmaceutically acceptable carrier, excipient or diluent.

21. The pharmaceutical composition of claim 20, further comprising an additional therapeutic agent selected from the group consisting of a chemotherapeutic or anti-proliferative agent, an anti-inflammatory agent, an immunomodulatory or immunosuppressive agent, a neurotrophic factor, an agent for treating cardiovascular disease, an agent for treating diabetes, and an agent for treating immunodeficiency disorders.

22. A compound according to any one of claims 1 to 14 for use in the treatment of an inflammatory or autoimmune disease.

23. A compound as claimed in any one of claims 1 to 14 for use in the treatment of any one of the conditions mentioned in claims 21 to 25.

24. Use of a compound according to any one of claims 1 to 14 in the manufacture of a medicament for the treatment of an inflammatory or autoimmune disease.

25. The invention as hereinbefore described.