CN113735859A - Kinase inhibitor - Google Patents

Abstract

The present invention relates to a kinase inhibitor comprising a compound of formula (I) or a pharmaceutically acceptable salt, solvate, ester, acid, metabolite or prodrug thereof. The invention also relates to pharmaceutical compositions comprising the kinase inhibitors, and uses and methods of using these compounds and compositions to inhibit JAK1, JAK2, JAK3, TYK2 kinase activity in a cell or subject, and/or to prevent or treat a condition associated with JAK1, JAK2, JAK3, TYK2 activity in a subject.

Description

Kinase inhibitor

Technical Field

The present application relates to a kinase inhibitor, and methods and uses for inhibiting kinase activity using such kinase inhibitors. More specifically, the invention relates to inhibitors capable of inhibiting JAK1, JAK2, JAK3, TYK2 kinase activity.

Background

Cytokines have important functions in regulating many aspects of immunity and inflammation. Ranging from the development and differentiation of immune cells to the suppression of immune responses. Type I and type II cytokine receptors lack intrinsic enzymatic activity capable of mediating signal transduction and therefore require binding to tyrosine kinases for this purpose. The JAK kinase family includes four distinct members, JAK1, JAK2, JAK3and TYK2, which bind to type i and type II cytokine receptors to control signal transduction (murrayppj, (2007). The JAK-STAT signalling pathway. j Immunol,178: 2623). Each JAK kinase is selective for receptors for certain cytokines. In this regard, JAK-deficient cell lines and mice have demonstrated an important role for each JAK protein in receptor signaling: JAK1 among the type II cytokine receptors (IFN and IL-10 family) that share the gp130 chain (IL-6 family) and the common Y chains (IL-2, IL-4, IL-7, IL-9, IL-15 and IL-21) (Rodig et al (1998) dispersion of the JAK1L genes antagonists and non-essential alcohols of the JAKs in cell-induced biological responses. Cel1,93: KL; Guschin et al (1995) A major Kinase for the protein type Kinase in JAK/STAT reaction in tissue reaction to interference protein-6. EMBO JJ.14: 1; British et al (K-expression) Kinase J.799: expression of protein expression J.15. expression of protein expression J.1); JAK2 in hematopoietic factors (Epo, Tpo, GM-CSF, IL-3, IL-5) and type II IFN (Parganas et al (1998). JAK2 is scientific for signalling through a _ variety of cytokine receptors. Cel1,93: 385); JAK3 is among the receptors sharing the common Y chain (IL-2 family) (Park et al, (1995). development of 1ymphoid cells in JAK3 kinase-specification die.Immunity, 3: 771; Thomis et al, (1995). Defects in B1 ymphocyte activation and Tlympocyte activation in die lacking JAK3.science,270: 794; Russell et al, (1995). Mution of JAK 3in a partial with SCID: examination of JAK 3in 1 lyphoid development. science,270: 797); and type 2 among receptors for IL-12, IL-23, IL-13, and type II IFNs (Karaghioff et al, (2000). Partial immunity in type K2-specific microorganism. immunity,13: 549; Shimoda et al, (2000) type K2 plant a restricted roll in IFNg signaling, throughput is requested for IL-1L 2-partitioned T cell function. immunity,13: 561; Mineishi et al, (2006) Human Tyrosiness kinase 2 restriction release reaction sites requirement in multiple cells immunity in cell synthesis, 745. III).

Upstream protein phosphorylation causes JAK activation, which in turn leads to downstream protein STAT recruitment, activation, dimerization. STAT dimers then function as transcription factors, translocating to the nucleus and activating transcription of a variety of responsive genes. There are seven identified STAT proteins: STAT1, STAT2, STAT3, STAT4, STAT5a, STAT5b, and STAT 6. Each specific cytokine receptor preferably binds to a specific STAT protein. Some binding is independent of cell type (e.g., IFNg-STAT1), while other binding may be associated with cell type (Murray PJ, (2007). The JAK-STATS identification pathway: input and output integration. J Imnunol,178: 2623).

The phenotype of deficient mice has provided insight into the function of each JAK and the cytokine receptors that signal transduction by that JAK. JAK3 binds exclusively to the common Y chain of receptors for IL-2, IL-4, IL-7, IL-9, IL-15, and IL-21 cytokines. Due to this specific binding, JAK3 knockout mice have the same phenotype as common Y chain deficient mice (Thomis et al, (1995). Defects in B1 phosphor activation and T1 phosphor activation in peptide lacking JAK3.science,270: 794; DiSanto et al, (1995). lysine depletion in mice with a targeted deletion of the interleukin 2receptor gamma chain. PNAS,92: 377). Furthermore, this phenotype is largely common in SCID patients with mutations/defects in the common Y chain or JAK3 gene (O' Shell et al, (2004). JAK3and the pathogenesis of segment combined immunity. mol Immunol,41: 727). JAK3-deficient mice survived but exhibited abnormal production of lymphocytes, which resulted in a reduction in size of the thymus (wild-type 1/100-1/10). JAK3-deficient peripheral T cells are unresponsive and have an activated memory cell phenotype (Baird et al, (1998). T cell depletion and activation in JAK 3-specific mice.J.Leuk.biol.63: 669). The thymus deficiency in these Mice is very similar to that seen in IL-7 and TIL-7 receptor Gene knockout Mice, suggesting that knock-out of IL-7 signaling may cause JAK3 mouse deficiency (von Freeden-Jeffry et al (1995). Lymphopenia in Interleukin (IL) -7Gene-deleted Mice identities IL-7as a non-reduced Cytokin. J. butyl Exp Med, 181: 1519; Peschon et al (1994). Early lymphocyte expansion is section amplified in Interleukin 7 applicator-specification. butyl Exp Med,180: 1955). Similar to SCID mice, these mice were free of NK cells, which could be attributed to the absence of IL-15 signaling (IL-15 signaling is a survival factor for these cells). Unlike SCID mice, JAK3 knock-out mice show defective B cell lymphopoiesis, whereas in human patients B cells are present in the circulation but do not respond, leading to hypoglobulinemia (O' Shea et al, (2004). JAK3and the pathogenesis of polypeptide combined immunology. mol immunol,41: 727). The explanation for this is that there are species-specific differences in the function of IL-7 in BE and T cell development in mice as well as humans. On the other hand, Grossman et al (1999. differentiated myelogenesis in mice lacking JAK3.blood,94:932:939) have shown that the loss of JAK 3in the T cell compartment contributes to the expansion of bone-mediated lines, leading to Dysregulated myelogenesis.

JAK 2-deficient mice are lethal at embryonic time due to the absence of committed erythropoiesis. Myeloid progenitor cells fail to respond to Epo, Tpo, IL-3, or GM-CSF, while G-CSF as well as IL-6 signaling are unaffected. JAK2 is not required for the generation, expansion or functional differentiation of lymphoid progenitors (Parganas et al (1998). JAK2 is scientific for signalling through a variety of cytokine receptors. Cel1,93: 385). JAK 1-deficient mice die perinatally due to lactation deficiency. JAK1 binds exclusively to gp130 chain common to the IL-6 cytokine family (i.e., LIF \ CNTF, OSM \ CT-1) and shares a fundamental component of receptors for the common y chain with JAK3 by binding to an unshared receptor subunit. In this regard, JAK 1-deficient mice display a similar hematopoiesis defect as JAK3-deficient mice. In addition, they show defective responses to neurotrophic factors as well as to all interferons (type II cytokine receptors) (Rodig et al, (1998) irradiation of the JAK1L gene antagonists and non-safedants of the JAKs in cytokine-induced biological responses, Cell1,93: 37).

Finally, mice deficient in Tyk2 showed a reduced response to IL-12 as well as IL-23 and only a Partial reduction in response to IFN-a (Karaghioff et al, (2000). Partial interference of cytokine responses in Tyk 2-specific microorganism immunity,13: 549; Shimoda et al, (2000). Typk 2 strain a structured roll in IFNg amplification, and an aqueous response for IL-12-mediated T cell function. immunity,13: 561). However, Human Tyk2 is deficient in demonstrating that Tyk2 is involved in signaling from IFN-a, IL-6, IL-10, IL-12, and IL-23 (Minegishi et al, (2006.) Human Tyrosine kinase 2 specific responses in multiple cell signals in immune, Immunity,25: 745).

The role of JAK kinases in transducing the signals of a large number of cytokines makes them potential targets for the treatment of diseases in which cytokines have pathogenic effects, such as inflammatory diseases, including, but not limited to, allergy and asthma, Chronic Obstructive Pulmonary Disease (COPD), bovine dermatoma, autoimmune diseases such as rheumatoid arthritis (rhamatoid arthritis) amyotrophic lateral sclerosis (amyotrophic lateral sclerosis) and multiple sclerosis c (multiple sclerosis), uveitis, transplant rejection, and solid and hematologic malignancies such as myeloproliferative disorders (myeloproliferative disorders), leukemia, and lymphoma.

In rheumatoid arthritis, an imbalance between the activity of proinflammatory cytokines and the activity of anti-inflammatory cytokines contributes to the induction of autoimmunity, followed by chronic inflammation and tissue destruction. In this regard, the pathogenic role of IL-6 in rheumatoid arthritis (CRA) has been clinically demonstrated using the anti-IL-6R antibody Tulipizumab (tocilizumab). IL-6 activates the transcription factor STAT3 Heinrich et al, (2003). Principles of Interleukin (IL) -6-type _ cytokine signaling and its regulation. biochem J.374:1) by using JAK1 bound to the gp130 receptor chain. Constitutive STAT3 mediates abnormal growth and survival properties of RA synovial cells (CIvashkiv et al (2003). The JAK/ZSTAT pathway in rheumatoid arthritis: pathogenic of protective Arth & Rheum.48: 2092). Other cytokines involved in the pathogenesis of arthritis include: IL-12 and ITIL-23, which are associated with Thl and Thl7 cell proliferation, respectively; IL-15 and GM-CSF (McInnes and Schett, (2007). Cytokines in the pathogenesis of rhematoid arthritis. Nature Rew Imnuol.7: 429.). Receptors for these cytokines also use JAK proteins for signal transduction, making JAK inhibitors a potential pleiotropic drug in this pathology. Thus, administration of JAK inhibitors, such as JAK inhibitors, in animal models of murine collagen-induced arthritis and rat adjuvant-induced arthritis, can reduce the incidence of inflammation and tissue destruction (Milici et al, (2008).

Inflammatory Bowel Disease (IBD) includes two major forms of enteritis: ulcerative colitis (ulcerogenic colitis) and crohn's disease. There is increasing evidence that various cytokines, including interleukins and interferons, have been implicated in The pathogenesis of IBD (Strober et al, (2002). The immunology of mucosal models of infllamation. Annu Rev Immunol.20: 495). Activation of the IL-6/ZSTAT3 cascade in lamina propria (laninacopia) T cells has been shown to induce long-term survival of pathogenic T cells (CATreya et al, (2000). Block of interfacial 6trans signaling supressants T-cell resistance against infection in viral expression. identification in Crohn's disease and experimental sensitivity in vivo. Nature Med.6: 583). In particular, STAT3 has been shown to have Constitutive activity in intestinal T cells in Crohn's disease patients, and JAK inhibitors have been shown to block Constitutive activation of STAT 3in these cells (Lovato et al, (2003). These observations indicate that the JAK-STAT pathway plays a pathogenic role in IBD and that JAK inhibitors may be therapeutic in this environment. Multiple sclerosis is an autoimmune demyelinating disease characterized by the formation of sympathetic blocks in the white matter. The role of cytokines in the development of multiple sclerosis has long been known. Potential therapies include blockade of IFN-g, IL-12, and IL-23(Steinman L. (2008.) Nuanced roles of cytokines in the gene major human prain disorders.J. Clin invest.118:3557), cytokines that signal through the JAK-STAT pathway. The use of Tyrphostin (JAK inhibitors) has been shown to inhibit IL-12 induced STAT3 phosphorylation and to reduce the incidence and severity of active and passive Experimental Autoimmune Encephalitis (EAE) (Bright et al (1999) Tyrphostin B42 inhibitor IL-12-induced tyrosination phosphorylation and activation of Janus kinase-2and present experimental allogenic encephalyisis, Imnuno. 162: 6255). Another multi-kinase inhibitor, CEP701, has been shown to reduce the secretion of TNF-a, IL-6 and IL-23 and to decrease the levels of phosphorylated STAT1, STAT 3and STAT5 in peripheral DCs of mice with EAE, thereby significantly improving the clinical course of EAE in mice (CSkarica et al, (2009). Signal transduction inhibition of APCs diagnostics, Thl7 and Thl tespones in experimental autoimmunity encephalyitis. J.Immunol.182: 4192.).

Psoriasis is an inflammatory disease of the skin that involves a process of infiltration and activation of immune cells that terminates in epithelial remodeling (epithelial remodelling). Current theory exploring the causes of psoriasis indicates that there is a subset of cytokines that control the interaction between the immune and epithelial cells (CNickoloffBJ (2007). Cracking the cytokine code in psisiasis, Nat Med,13: 2420). In this regard, IL-23 produced by dendritic cells was found to increase with IL-12 in psoriatic skin. IL-23 induces the formation of Thl7 cells, which in turn produce IL-17 and IL-22, the latter responsible for epidermal thickening. IL-23 and IL-22 induce phosphorylation of STAT-3, which is abundantly present in bovine skin. Therefore, JAK inhibitors may be therapeutic in this environment. Thus, it has been found that in an idiopathic T cell dependent mouse model of psoriasis, a JAK173 inhibitor, R348, reduces bovine papilloma skin inflammation (CChang et al, (2009). JAK3 inhibition signaling inflammation in vitro inflammation on CD18 mutant PL/J mice. J Immunol1.183: 2183).

Hematologic malignancies in which the JAK-STAT pathway is dysregulated may benefit from JAK inhibition. Recent studies have shown that JAK2 kinase activity is deregulated in the areas of human myeloproliferative diseases, CIhIe and Gililand,2007 (including polycythemia vera), myelofibrosis (myelofibrosis) and essential thrombocytosis (stressed thrombocytosis), by chromosomal translocations and mutations in the pseudokinase domain (such as JAK2V617F mutations). in this regard, several JAK inhibitors have been proposed that effectively treat JAK2, such as TG-101209 (Paranani et al, (2007) TG101209, a small molecular JAK2-selective inhibition of bone defects, JAK 12V 617 and JAK 515L7/K tissue of JAK2V F and JAK 493W 515L7/K tissue defects 21:1658-68), TG101348 (Cell of JAK 31. 12. A) and JAK 31. 12, JAK 35, JAK 12 and JAK 12, and P31, III, and the like, (TG 201, JAK2V 2. cholesterol, JAK 3531, JAK 12, and P31, and 12. A, JAK 12. A, JAK2, a Blood,111:5663), CP-690,550(Manshouri et al, (2008) The JAK kinase inhibitor CP-690,550 side reactions The growth of human multicyteria cell carring The JAK2V617F mutation. cancer Sci,99:1265) and CYT387 (Paranani et al, (2009) CYT387, a _ selective JAK1/JAK2 inhibitor of The kinase selection and peclinic standards using cells and primary cells 617F for The treatment of myeloproliferative diseases based on their mutated V617 activity. Similarly, T cell leukemia caused by transformation of human leukemia virus CHTLV-1) is associated with constitutive activation of JAK3and STAT5 (Migone et al (1995). Constituteiveactivated JAK-STAT pathway in T cells transformed with HTLV-I.science,269: 79). JAK inhibitors may have a therapeutic effect in this context (CTomita et al, (2006). Inhibition of connective active JAK-STAT pathway responses _ cell growth of human T-cell leukemia virus type TI-induced T-cell lines and primary addition T-cell leukemia cells.retrovirology,3: 22). The activation of the JAK1 mutation can cause acute lymphoblastoma and leukemia (Flex et al, (2008). Somatically acquired JAK1 mutations in adult access lymphoblast leukemia. J.exp. Med.205:751-8), indicating that the kinase can be used as a target for developing novel anti-leukemia drugs.

Conditions contemplated for the targeting of the JAK pathway or modulation of JAK kinases (especially JAK1, JAK2, JAK3and TYK2 kinases) for therapeutic or prophylactic treatment of disease include: neoplastic diseases (e.g., leukemia, lymphoma, etc.); transplant rejection, bone marrow transplantation applications (e.g., graft versus host disease); autoimmune diseases (e.g., diabetes, multiple sclerosis, rheumatoid arthritis, inflammatory bowel disease); inflammatory diseases of the respiratory tract (e.g., asthma, chronic obstructive pulmonary disease); inflammatory-related or allergic eye diseases (e.g. dry eye glaucoma, uveitis, diabetic retinopathy cdiabeta retinitis), allergic conjunctivitis or age-related macular degeneration) and inflammatory diseases of the skin (e.g. atopic dermatitis or psoriasis).

Given that many conditions are expected to benefit from treatment involving modulation of the JAK pathway or JAK kinases, it is apparent that novel compounds that modulate the JAK pathway and the use of these compounds should provide substantial therapeutic benefits to a variety of patients.

The present invention provides a novel kinase inhibitor compound for use in the treatment of conditions in which targeting of the JAK pathway or inhibition of JAK kinases may be therapeutically useful.

The invention mainly discovers an inhibitor which has a brand-new structure and has a strong inhibition effect on JAK kinase.

Disclosure of Invention

The present invention provides a selective kinase inhibitor comprising a compound of formula (I) or a pharmaceutically acceptable salt, solvate, ester, acid, metabolite or prodrug thereof,

wherein the content of the first and second substances,

x is a nitrogen atom or a carbon atom;

y is a nitrogen atom or a carbon atom;

z is a direct bond or-NH-;

R₁selected from hydrogen atoms, halogen atoms, C_1-6Alkyl, carboxamido;

R₂selected from the group consisting of optionally substituted by 1-3 independent R₃Phenyl, pyridyl, pyrazolyl or pyrimidinyl substituted with radicals;

R₃independently selected from halogen, amino, C_1-6Alkyl radical, C_1-6Haloalkyl, C_1-6Alkoxy, (4-methylpiperazin-1-yl) methyl, methoxy-substituted phenyl, naphthyl, 4-pyridyl, 3-piperidyl or 4-piperidyl optionally substituted by methyl.

A pharmaceutical composition comprising one or more kinase inhibitors as described above, together with a pharmaceutically acceptable carrier or excipient, and optionally other therapeutic agents, for use in the treatment of a disease, disorder or condition, wherein the disease, disorder or condition is selected from the following autoimmune diseases: arthritis, rheumatoid arthritis, osteoarthritis, lupus, rheumatoid arthritis, inflammatory bowel disease, psoriatic arthritis, osteoarthritis, still's disease, juvenile arthritis, diabetes, myasthenia gravis, hashimoto's thyroiditis, alder's thyroiditis, graves' disease, rheumatoid arthritis syndrome, multiple sclerosis, infectious neuronitis, acute disseminated encephalomyelitis, addison's disease, optic twin-myotic syndrome, ankylosing spondylitis, antiphospholipid antibody syndrome, aplastic anemia, autoimmune hepatitis, celiac disease, goodpasture's syndrome, idiopathic thrombocytopenic purpura, optic neuritis, scleroderma, primary biliary cirrhosis, reiter's syndrome, takayasu's arteritis, temporal arteritis, warm autoimmune hemolytic anemia, wegener's granulomatosis, inflammatory bowel disease, acute disseminated encephalomyelitis, addictitis encephalomyelitis, addictal encephalomyelitis, addictus disease, addison's disease, optic neuritis, optic neuropathy, myelitis obliqus syndrome, and other syndrome, myelitis obliqus syndrome, and other syndrome, Psoriasis, systemic alopecia, behcet's disease, chronic fatigue, familial autonomic dysfunction, endometriosis, interstitial cystitis, neuromuscular stiffness, scleroderma or vulvodynia.

Use of one or more kinase inhibitors as described above in the manufacture of a medicament for inhibiting the activity of JAK1, JAK2, JAK3, TYK2 kinase for treating a disease, disorder or condition, wherein the disease, disorder or condition is a JAK1, JAK2, JAK3, TYK2 kinase-associated disease.

Use of one or more kinase inhibitors as described above for the manufacture of a medicament for the treatment or prevention of a disease, disorder or condition associated with JAK1, JAK2, JAK3, TYK2 activity.

Detailed Description

Term(s) for

Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which the claimed subject matter belongs.

The present invention employs, unless otherwise indicated, conventional methods of mass spectrometry, NMR, HPLC, protein chemistry, biochemistry, recombinant DNA techniques and pharmacology, within the skill of the art. Unless a specific definition is provided, nomenclature and laboratory procedures and techniques related to the chemistry described herein, such as analytical chemistry, synthetic organic chemistry, and medical and pharmaceutical chemistry, are known to those skilled in the art. In general, the foregoing techniques and procedures may be practiced by conventional methods well known in the art and described in various general and more specific documents that are cited and discussed in this specification.

The term "alkyl" refers to an aliphatic hydrocarbon group, which may be a branched or straight chain alkyl group. Depending on the structure, the alkyl group may be a monovalent group or a divalent group (i.e., alkylene). In the present invention, the alkyl group is preferably an alkyl group having 1 to 8 carbon atoms, more preferably a "lower alkyl group" having 1 to 6 carbon atoms, and even more preferably an alkyl group having 1 to 4 carbon atoms. Typical alkyl groups include, but are not limited to, methyl, ethyl, propyl, butyl, pentyl, hexyl, and the like. It is to be understood that reference herein to "alkyl" includes reference to that alkyl in all configurations and conformations that may exist, for example reference herein to "propyl" includes n-propyl and isopropyl, "butyl" includes n-butyl, isobutyl and tert-butyl, "pentyl" includes n-pentyl, isopropyl, neopentyl, tert-pentyl, and pent-3-yl, and the like.

The term "alkoxy" refers to an-O-alkyl group, wherein alkyl is as defined herein. Typical alkoxy groups include, but are not limited to, methoxy, ethoxy, propoxy, butoxy, pentoxy, hexoxy, and the like.

The term "alkoxyalkyl" means an alkyl group, as defined herein, substituted with an alkoxy group, as defined herein.

The term "cycloalkyl" refers to a monocyclic or multicyclic group that contains only carbon and hydrogen. Cycloalkyl groups include groups having 3 to 12 ring atoms. Depending on the structure, the cycloalkyl group can be a monovalent group or a divalent group (e.g., cycloalkylene). In the present invention, the cycloalkyl group is preferably a cycloalkyl group having 3 to 8 carbon atoms, more preferably a "lower cycloalkyl group" having 3 to 6 carbon atoms. Examples of cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclopentenyl, cyclohexenyl, cycloheptenyl, and adamantyl.

The term "alkyl (cycloalkyl)" or "cycloalkylalkyl" means that an alkyl group, as defined herein, is substituted with a cycloalkyl group, as defined herein. Non-limiting cycloalkylalkyl groups include cyclopropylmethyl, cyclobutylmethyl, cyclopentylmethyl, cyclohexylmethyl, and the like.

The term "aryl" refers to a planar ring having a delocalized pi-electron system and containing 4n +2 pi electrons, where n is an integer. An aryl ring may be composed of five, six, seven, eight, nine or more than nine atoms. The aryl group may be optionally substituted. The term "aryl" includes carbocyclic aryl (e.g., phenyl) and heterocyclic aryl (or "heteroaryl") groups (e.g., pyridine). The term includes monocyclic or fused-ring polycyclic (i.e., rings that share adjacent pairs of carbon atoms) groups.

The term "aryl" as used herein means that each of the ring-forming atoms in the aromatic ring is a carbon atom. The aryl ring may be composed of five, six, seven, eight, nine or more than nine atoms. The aryl group may be optionally substituted. Examples of aryl groups include, but are not limited to, phenyl, naphthyl, phenanthryl, anthracyl, fluorenyl, and indenyl. Depending on the structure, the aryl group can be a monovalent group or a divalent group (i.e., arylene).

The term "aryloxy" refers to-O-aryl, wherein aryl is as defined herein.

The term "heteroaryl" refers to an aryl group that includes one or more ring heteroatoms selected from nitrogen, oxygen, and sulfur. An N-containing "heteroaryl" moiety means that at least one of the backbone atoms in the ring in the aryl group is a nitrogen atom. Depending on the structure, heteroaryl groups may be monovalent or divalent (i.e., heteroarylene). Examples of heteroaryl groups include, but are not limited to, pyridyl, imidazolyl, pyrimidinyl, pyrazolyl, triazolyl, pyrazinyl, tetrazolyl, furyl, thienyl, isoxazolyl, thiazolyl, oxazolyl, isothiazolyl, pyrrolyl, quinolyl, isoquinolyl, indolyl, benzimidazolyl, benzofuranyl, indazolyl, indolizinyl, phthalazinyl, pyridazinyl, isoindolyl, pteridinyl, purinyl, oxadiazolyl, thiadiazolyl, furazanyl, benzofurazanyl, benzothienyl, benzothiazolyl, benzoxazolyl, quinazolinyl, naphthyridinyl, and furopyridyl, and the like.

The term "alkyl (aryl)" or "aralkyl" means that an alkyl group, as defined herein, is substituted with an aryl group, as defined herein. Non-limiting alkyl (aryl) groups include benzyl, phenethyl, and the like.

The term "alkyl (heteroaryl)" or "heteroarylalkyl" means that an alkyl group, as defined herein, is substituted with a heteroaryl group, as defined herein.

The term "heteroalkyl," as used herein, means that one or more backbone chain atoms in an alkyl group, as defined herein, is a heteroatom, such as oxygen, nitrogen, sulfur, silicon, phosphorus, or combinations thereof. The heteroatom(s) may be located anywhere within the heteroalkyl group or at the position where the heteroalkyl group is attached to the remainder of the molecule.

The term "heterocycloalkyl" or "heterocyclyl" as used herein means that one or more of the ring-forming atoms in the non-aromatic ring is a heteroatom selected from nitrogen, oxygen and sulfur. A heterocycloalkyl ring can be composed of three, four, five, six, seven, eight, nine, or more than nine atoms. The heterocycloalkyl ring may be optionally substituted. Examples of heterocycloalkyl groups include, but are not limited to, lactams, lactones, cyclic imines, cyclic thioimines, cyclic carbamates, tetrahydrothiopyrans, 4H-pyrans, tetrahydropyrans, piperidines, 1, 3-dioxins, 1, 3-dioxanes, 1, 4-dioxins, 1, 4-dioxanes, piperazines, 1, 3-oxathianes, 1, 4-oxathianes, tetrahydro-1, 4-thiazines, 2H-1, 2-oxazines, maleimides, succinimides, barbituric acid, thiobarbituric acid, dioxopiperazines, hydantoins, dihydrouracils, morpholines, trioxanes, hexahydro-1, 3, 5-triazines, tetrahydrothiophenes, tetrahydrofurans, pyrrolines, pyrrolidines, imidazolidines, pyrrolidones, and the like, Pyrazoline, pyrazolidine, imidazoline, imidazolidine, 1, 3-dioxole, 1, 3-dioxolane, 1, 3-dithiole, 1, 3-dithiolane, isoxazoline, isoxazolidine, oxazoline, oxazolidine, oxazolidinone, thiazoline, thiazolidine, and 1, 3-oxathiolane. Depending on the structure, heterocycloalkyl groups can be monovalent or divalent (i.e., heterocycloalkylene).

The term "alkyl (heterocycloalkyl)" or "heterocycloalkylalkyl" means that an alkyl group, as defined herein, is substituted with a heterocycloalkyl group, as defined herein.

The term "alkoxy (heterocycloalkyl)" or "heterocycloalkylalkoxy" means that an alkoxy group, as defined herein, is substituted with a heterocycloalkyl group, as defined herein.

The term "halo" or "halogen" refers to fluorine, chlorine, bromine and iodine.

The terms "haloalkyl", "haloalkoxy" and "haloheteroalkyl" include alkyl, alkoxy, or heteroalkyl groups in which at least one hydrogen is replaced with a halogen atom. In certain embodiments, if two or more hydrogen atoms are replaced with a halogen atom, the halogen atoms are the same or different from each other.

The term "hydroxy" refers to an-OH group.

The term "cyano" refers to the group — CN.

The term "ester group" refers to a chemical moiety having the formula-COOR, wherein R is selected from the group consisting of alkyl, cycloalkyl, aryl, heteroaryl (attached through a ring carbon), and heterocyclyl (attached through a ring carbon).

The term "amino" refers to the group-NH₂A group.

The term "aminoacyl" refers to-CO-NH₂A group.

The term "amido" or "amido" refers to the group-NR-CO-R ', wherein R and R' are each independently hydrogen or alkyl.

The term "alkylamino" refers to an amino substituent further substituted by one or two alkyl groups, in particular to the group-NRR ', wherein R and R ' are each independently selected from hydrogen or lower alkyl, with the proviso that-NRR ' is not-NH₂. "alkylamino" includes the group-NH-thereof₂The nitrogen of (a) is linked to at least one alkyl group. Examples of alkylamino groups include, but are not limited to, methylamino, ethylamino, and the like. "dialkylamino" includes wherein-NH₂The nitrogen of (a) is linked to at least two other alkyl groups. Examples of dialkylamino groups include, but are not limited to, dimethylamino, diethylamino, and the like.

The terms "arylamino" and "diarylamino" refer to an amino substituent further substituted with one or two aryl groups, specifically the group-NRR ', where R and R' are each independently selected from hydrogen, lower alkyl, or aryl, where N is attached to at least one or two aryl groups, respectively.

The term "cycloalkylamino" refers to an amino substituent further substituted with one or two cycloalkyl groups as defined herein.

The term "heteroalkylamino" refers to an amino substituent further substituted with one or two heteroalkyl groups as defined herein.

The term "aralkylamino" as used herein refers to the group-NRR 'wherein R is loweraralkyl and R' is hydrogen, loweralkyl, aryl or loweraralkyl.

The term "heteroarylamino" refers to an amino substituent further substituted with one or two heteroaryl groups as defined herein.

The term "heterocycloalkylamino" means that an amino group, as defined herein, is substituted with a heterocycloalkyl group, as defined herein.

The term "alkylaminoalkyl" means that an alkyl group, as defined herein, is substituted with an alkylamino group, as defined herein.

The term "aminoalkyl" refers to an alkyl substituent further substituted with one or more amino groups.

The term "aminoalkoxy" refers to an alkoxy substituent further substituted with one or more amino groups.

The term "hydroxyalkyl" or "hydroxyalkyl" refers to an alkyl substituent further substituted with one or more hydroxyl groups.

The term "cyanoalkyl" refers to an alkyl substituent further substituted with one or more cyano groups.

The term "acyl" refers to the monovalent radical remaining after removal of the hydroxyl group by an organic or inorganic oxoacid, and has the general formula R-M (O) -where M is typically C.

The term "carbonyl" is an organic functional group consisting of two atoms, carbon and oxygen, connected by a double bond (C ═ O).

The term "alkanoyl" or "alkylcarbonyl" refers to a carbonyl group further substituted with an alkyl group. Typical alkanoyl groups include, but are not limited to, acetyl, propionyl, butyryl, pentanoyl, hexanoyl, and the like.

The term "arylcarbonyl" means a carbonyl group, as defined herein, substituted with an aryl group, as defined herein.

The term "alkoxycarbonyl" refers to a carbonyl group further substituted with an alkoxy group.

The term "heterocycloalkylcarbonyl" refers to a carbonyl group further substituted with a heterocycloalkyl group.

The terms "alkylaminocarbonyl", "cycloalkylaminocarbonyl", "arylaminocarbonyl", "aralkylaminocarbonyl", "heteroarylaminocarbonyl", respectively, mean that the carbonyl group, as defined herein, is substituted with alkylamino, cycloalkylamino, arylamino, aralkylamino, or heteroarylamino, respectively, as defined herein.

The term "alkylcarbonylalkyl" or "alkanoylalkyl" refers to an alkyl group further substituted with one alkylcarbonyl group.

The term "alkylcarbonylalkoxy" or "alkanoylalkoxy" refers to an alkoxy group further substituted with one alkylcarbonyl group.

The term "heterocycloalkylcarbonylalkyl" refers to an alkyl group further substituted with a heterocycloalkylcarbonyl group.

The term "mercapto" refers to the-SH group. The term "alkylthio" means a mercapto group, as defined herein, substituted with an alkyl group, as defined herein.

The term "sulfonyl" or "sulfonyl" refers to a functional group of a sulfonic acid after loss of the hydroxyl group, specifically — S (═ O)₂-a group.

The term "sulfoxido" or "sulfinyl" refers to-S (═ O) -.

The term "aminosulfonyl" or "aminosulfonyl" refers to-S (═ O)₂-NH₂A group.

The term "alkylsulfinyl" or "alkylsulfinyl" refers to — S (═ O) -R, where R is alkyl.

The term "alkylsulfonyl" or "alkylsulfonyl" refers to-S (═ O)₂-R, wherein R is alkyl.

The term "alkylaminosulfonyl" means that a sulfone group, as defined herein, is substituted with an alkylamino group, as defined herein.

The term "alkylsulfonylamino" or "cycloalkylsulfonylamino" means that the amino group as defined herein is substituted with an alkylsulfonyl or cycloalkylsulfonyl group as defined herein.

The terms "cycloalkylsulfonyl" and "cycloalkylsulfonyl" refer to-S (═ O)₂-R, wherein R is cycloalkyl.

The terms "alkylsulfonamide" and "cycloalkylsulfonamide" refer to-NH-S (═ O)₂-R, wherein R is alkyl and cycloalkyl, respectively.

The term "quaternary ammonium group" means-N⁺RR 'R' where R, R 'and R' are each independently selected from alkyl groups having 1-8 carbon atoms.

The term "optionally" means that one or more of the subsequently described events may or may not occur, and includes both occurring events and non-occurring events. The term "optionally substituted" or "substituted" means that the referenced group may be substituted with one or more additional groups each and independently selected from alkyl, cycloalkyl, aryl, heteroaryl, heterocyclyl, hydroxy, alkoxy, cyano, halo, amido, nitro, haloalkyl, amino, methanesulfonyl, alkylcarbonyl, alkoxycarbonyl, heteroarylalkyl, heterocycloalkylalkyl, aminoacyl, amino protecting groups, and the like. Among them, the amino-protecting group is preferably selected from pivaloyl, t-butoxycarbonyl, benzyloxycarbonyl, 9-fluorenylmethoxycarbonyl, benzyl, p-methoxybenzyl, allyloxycarbonyl, trifluoroacetyl and the like.

The term "Tyrosine Protein Kinase (TPK)" as used herein is a class of kinases that catalyze the transfer of gamma-phosphate from ATP to protein tyrosine residues, which catalyze the phosphorylation of tyrosine residues of various substrate proteins, and play an important role in cell growth, proliferation, and differentiation.

The term "inhibition", "inhibitory" or "inhibitor" of a kinase, as used herein, means that phosphotransferase activity is inhibited.

A "metabolite" of a compound disclosed herein is a derivative of the compound that is formed when the compound is metabolized. The term "active metabolite" refers to a biologically active derivative of a compound that is formed when the compound is metabolized. The term "metabolized" as used herein, refers to the sum of processes (including, but not limited to, hydrolysis reactions and reactions catalyzed by enzymes, e.g., oxidation reactions) by which a particular substance is altered by an organism. Thus, enzymes can produce specific structural transformations into compounds. For example, cytochrome P450 catalyzes a variety of oxidation and reduction reactions, while phosphoglucose glycyltransferase catalyzes the conversion of activated glucuronic acid molecules to aromatic alcohols, aliphatic alcohols, carboxylic acids, amines and free sulfhydryl groups. Further information on metabolism can be obtained from The pharmaceutical Basis of therapeutics, ninth edition, McGraw-Hill (1996). Metabolites of the compounds disclosed herein can be identified by administering the compounds to a host and analyzing a tissue sample from the host, or by incubating the compounds with hepatocytes in vitro and analyzing the resulting compounds. Both methods are known in the art. In some embodiments, the metabolite of the compound is formed by an oxidation process and corresponds to the corresponding hydroxyl-containing compound. In some embodiments, the compound is metabolized to a pharmaceutically active metabolite. The term "modulate," as used herein, refers to interacting, directly or indirectly, with a target to alter the activity of the target, including by way of example only, enhancing the activity of the target, inhibiting the activity of the target, limiting the activity of the target, or prolonging the activity of the target.

The term "target protein" as used herein refers to a protein molecule or portion of a protein that can be bound by a selective binding compound. In certain embodiments, the target protein is tyrosine kinase KIT (wild-type or various mutations or combinations thereof), ABL (wild-type or various mutations or combinations thereof), EGFR (wild-type or various mutations or combinations thereof), FLT3 (wild-type or various mutations or combinations thereof), VEGFR2 (wild-type or various mutations or combinations thereof), RET (wild-type or various mutations or combinations thereof), pdgfra (wild-type or various mutations or combinations thereof), PDGFR β (wild-type or various mutations or combinations thereof), BCR/ABL (wild-type or various mutations or combinations thereof), FGFR1 (wild-type or various mutations or combinations thereof), FGFR2 (wild-type or various mutations or combinations thereof), FGFR3 (wild-type or various mutations or combinations thereof), FGFR4 (wild-type or various mutations or combinations thereof).

IC as used herein₅₀Refers to the amount, concentration or dose of a particular test compound that achieves 50% inhibition of the maximal effect in an assay that measures such effect.

EC as used herein₅₀Refers to a dose, concentration, or amount of a test compound that elicits a dose-dependent response that is 50% of the maximal expression of a particular response that a particular test compound induces, stimulates, or potentiates.

GI as used herein₅₀Is meant to refer to the concentration of drug required to inhibit 50% of the cell growth, i.e., the concentration of drug at which 50% of the cell (e.g., cancer cell) growth is inhibited or controlled by the drug.

The novel kinase inhibitors of the present invention

The present invention provides a novel kinase inhibitor comprising a compound of formula (I) or a pharmaceutically acceptable salt, solvate, ester, acid, metabolite or prodrug thereof,

wherein the content of the first and second substances,

x is a nitrogen atom or a carbon atom;

y is a nitrogen atom or a carbon atom;

z is a direct bond or-NH-;

R₁selected from hydrogen atoms, halogen atoms, C_1-6Alkyl or carboxamido;

R₂selected from the group consisting of optionally substituted by 1-3 independent R₃Phenyl, pyridyl, pyrazolyl or pyrimidinyl substituted with radicals;

R₃independently selected from halogen, amino, C_1-6Alkyl radical, C_1-6Haloalkyl, C_1-6Alkoxy, (4-methylpiperazin-1-yl) methyl, methoxy-substituted phenyl, naphthyl, 4-pyridyl, 3-piperidyl or 4-piperidyl optionally substituted by methyl.

For each variable, any combination of the above groups is also contemplated herein. It can be understood that: substituents and substitution patterns on the compounds provided herein can be selected by one of skill in the art to provide compounds that are chemically stable and can be synthesized using techniques known in the art as well as those set forth herein.

In a preferred embodiment, the inhibitors of the present invention comprise the compounds of table 1 below, or a pharmaceutically acceptable salt, solvate, ester, acid, metabolite or prodrug thereof.

TABLE 1

Described herein are novel kinase inhibitors. Pharmaceutically acceptable salts, solvates, esters, acids, pharmaceutically active metabolites and prodrugs of this compound are also described herein.

In additional or further embodiments, the compounds described herein are metabolized in vivo upon administration to an organism in need thereof to produce a metabolite, which is then used to produce a desired effect, including a desired therapeutic effect.

The compounds described herein may be formulated and/or used as pharmaceutically acceptable salts. Types of pharmaceutically acceptable salts include, but are not limited to: (1) acid addition salts, formed by reacting the free base form of the compound with pharmaceutically acceptable inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, metaphosphoric acid, and the like; or with an organic acid such as acetic acid, propionic acid, hexanoic acid, cyclopentanepropionic acid, glycolic acid, pyruvic acid, lactic acid, malonic acid, malic acid, citric acid, succinic acid, maleic acid, tartaric acid, fumaric acid, trifluoroacetic acid, benzoic acid, 3- (4-hydroxybenzoyl) benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, 1, 2-ethanedisulfonic acid, 2-hydroxyethanesulfonic acid, benzenesulfonic acid, toluenesulfonic acid, 4-methylbicyclo- [2.2.2] oct-2-ene-1-carboxylic acid, 2-naphthalenesulfonic acid, tert-butylacetic acid, glucoheptonic acid, 4' -methylenebis- (3-hydroxy-2-ene-1-carboxylic acid), 3-phenylpropionic acid, trimethylacetic acid, or mixtures thereof, Dodecyl sulfuric acid, gluconic acid, glutamic acid, salicylic acid, hydroxynaphthoic acid, stearic acid, muconic acid, and the like; (2) base addition salts, which are formed when an acidic proton in the parent compound is replaced by a metal ion, such as an alkali metal ion (e.g., lithium, sodium, potassium), alkaline earth metal ion (e.g., magnesium or calcium), or aluminum ion; or coordinating with organic base or inorganic base, and acceptable organic base includes ethanolamine, diethanolamine, triethanolamine, trimethylamine, N-methylglucamine, etc.; acceptable inorganic bases include aluminum hydroxide, calcium hydroxide, potassium hydroxide, sodium carbonate, sodium hydroxide, and the like.

The corresponding counterion of the pharmaceutically acceptable salt can be analyzed and identified using a variety of methods including, but not limited to, ion exchange chromatography, ion chromatography, capillary electrophoresis, inductively coupled plasma, atomic absorption spectroscopy, mass spectrometry, or any combination thereof.

Recovering the salt using at least one of the following techniques: filtration, precipitation with a non-solvent followed by filtration, evaporation of the solvent, or in the case of aqueous solutions, lyophilization.

Screening and characterization of pharmaceutically acceptable salts, polymorphs, and/or solvates can be accomplished using a variety of techniques including, but not limited to, thermal analysis, X-ray diffraction, spectroscopy, microscopy, elemental analysis. Various spectroscopic techniques used include, but are not limited to, Raman, FTIR, UVIS, and NMR (liquid and solid state). Various microscopy techniques include, but are not limited to, IR microscopy and Raman (Raman) microscopy.

Pharmaceutical compositions of the invention

The present application also provides pharmaceutical compositions comprising at least one compound of formula (I) or a pharmaceutically acceptable salt, solvate, ester, acid, pharmaceutically active metabolite or prodrug of said compound, and a pharmaceutically acceptable carrier or excipient, and optionally other therapeutic agents.

During the course of treatment, the agent may be used alone or in combination with one or more other therapeutic agents, as the case may be. A medicament comprising a compound of the invention may be administered to a patient by at least one of injection, oral, inhalation, rectal and transdermal administration. The additional therapeutic agent may be selected from the following: immunosuppressants (e.g. tacrolimus, cyclosporin, rapamycin, methotrexate, cyclophosphamide, azathioprine, mercaptopurine, mycophenolate mofetil or FTY720), glucocorticoids (e.g. prednisone, cortisone acetate, prednisolone, methylprednisolone, dexamethasone, betamethasone, triamcinolone, prednisolone, beclomethasone, fludrocortisone acetate, deoxycorticosterone acetate, aldosterone), non-steroidal anti-inflammatory drugs (e.g. salicylates, aryl alkanoic acids, 2-aryl propionic acids, N-aryl anthranilic acids, oxicams, coxib or sulphanilides), allergy vaccines, antihistamines, anti-leukotrienes, β -agonists, theophylline, anticholinergic drugs or other selective kinase inhibitors (e.g. mTOR inhibitors, c-Met inhibitors) or her2 antibody-drugs. In addition, other therapeutic agents mentioned may also be Rapamycin (Rapamycin), Crizotinib (Crizotinib), tamoxifen, raloxifene, anastrozole, exemestane, letrozole, herceptin^TM(trastuzumab) Glibc^TM(imatinib), paclitaxel^TM(paclitaxel), cyclophosphamide, lovastatin, minocycline (Minosine), cytarabine, 5-fluorouracil (5-FU), Methotrexate (MTX), taxotere^TM(docetaxel), norrad^TM(goserelin), vincristine, vinblastine, nocodazole, teniposide, etoposide, and jiaojian^TM(gemcitabine), Epothilone (Epothilone), norben, camptothecin, daunorubicin (daunonibiin), dactinomycin, mitoxantrone, amsacrine, doxorubicin (doxorubicin), epirubicin or idarubicin. Alternatively, the other therapeutic agent may also be a cytokine such as G-CSF (granulocyte colony stimulating factor). Alternatively, the other therapeutic agent may also be, for example, but not limited to, CMF (cyclophosphamide, methotrexate, and 5-fluorouracil), CAF (cyclophosphamide, doxorubicin, and 5-fluorouracil), AC (doxorubicin and cyclophosphamide), FEC (5-fluorouracil, epirubicin, and cyclophosphamide), ACT or ATC (doxorubicin, cyclophosphamide, and paclitaxel), or CMFP (cyclophosphamide, methotrexate, 5-fluorouracil, and prednisone).

In embodiments of the invention, where a patient is treated according to the invention, the amount of a given drug will depend on factors such as the particular dosing regimen, the type of disease or disorder and its severity, the uniqueness (e.g., body weight) of the subject or host in need of treatment, however, the dosage administered may be routinely determined by methods known in the art depending on the particular circumstances, including, for example, the particular drug that has been employed, the route of administration, the disorder being treated, and the subject or host being treated. In general, for dosages used for adult human therapy, dosages administered will typically range from 0.02 to 5000 mg/day, for example from about 1 to 1500 mg/day. The desired dose may conveniently be presented as a single dose, or as divided doses administered simultaneously (or over a short period of time) or at appropriate intervals, for example two, three, four or more divided doses per day. It will be appreciated by those skilled in the art that, notwithstanding the dosage ranges set forth above, the specific effective amounts may be adjusted as appropriate to the circumstances of the patient and in conjunction with the diagnosis by the physician.

Use of the medicament of the invention

The kinase inhibitors of the present invention include at least one compound of formula (I) or a pharmaceutically acceptable salt, solvate, ester, acid, metabolite, or prodrug thereof, or a pharmaceutical composition for reducing or inhibiting JAK1, JAK2, JAK3, and TYK2 kinase activity in a cell or subject, and/or preventing or treating a disorder associated with JAK1, JAK2, JAK3, and TYK2 kinase activity in a subject.

A compound of formula (I) or a pharmaceutically acceptable salt, solvate, ester, acid, metabolite or prodrug thereof, or a pharmaceutical composition thereof, is useful for treating, preventing or ameliorating an autoimmune disease selected from the group consisting of: arthritis, rheumatoid arthritis, osteoarthritis, lupus, rheumatoid arthritis, inflammatory bowel disease, psoriatic arthritis, osteoarthritis, Still's disease, juvenile arthritis, diabetes, myasthenia gravis, Hashimoto's thyroiditis, oddmetitis, Graves ' disease, rheumatoid arthritis syndrome (rheumatoid arthritis syndrome)

syndrome), multiple sclerosis, infectious neuronitis (Guillain-Barr é syndrome), acute disseminated encephalomyelitis, Addison's disease, optic glans twin-myoclonus syndrome, ankylosing spondylitis, antiphospholipid antibody syndrome, aplastic anemia, autoimmune hepatitis, celiac disease, Goodpasture's syndrome, idiopathic thrombocytopenic purpura, optic neuritis, scleroderma, primary biliary cirrhosis, Retter's syndrome, Tayassus's teritis, temporal arteritis, warm autoimmune hemolytic anemia, Wegener's granulomatosis, psoriasis, systemic alopecia, Herceptive cuter's disease, chronic fatigue of the nerve, abnormal uterine function, cervical spondyloschisis, cervical spondylosis, chronic fatigue of the uterine system, chronic fatigue of the human body, chronic fatigue, Interstitial cystitis, neuromuscular stiffness, scleroderma or vulvodynia.

Preparation of the Compounds

The compounds of formula (I) may be synthesized using standard synthetic techniques known to those skilled in the art or using methods known in the art in combination with the methods described herein. In addition, the solvents, temperatures, and other reaction conditions set forth herein may be varied according to the skill in the art. As a further guide, the following synthesis method may be used.

The reactions may be used sequentially to provide the compounds described herein; or they may be used to synthesize fragments that are subsequently added by the methods described herein and/or known in the art.

In certain embodiments, provided herein are methods of making and methods of using the kinase inhibitor compounds described herein. In certain embodiments, the compounds described herein can be synthesized using the following synthetic schemes. The compounds can be synthesized using procedures analogous to those described below, by using appropriate alternative starting materials.

The starting materials for synthesizing the compounds described herein may be synthesized or may be obtained from commercial sources. The compounds described herein and other related compounds having different substituents can be synthesized using techniques and starting materials known to those skilled in the art. General methods of preparing the compounds disclosed herein can be derived from reactions known in the art, and the reactions can be modified by reagents and conditions deemed appropriate by those skilled in the art to incorporate various moieties in the molecules provided herein.

If desired, the reaction product may be isolated and purified using conventional techniques, including but not limited to filtration, distillation, crystallization, chromatography, and the like. These products can be characterized using conventional methods, including physical constants and profile data.

Non-limiting examples of synthetic schemes for preparing compounds of formula (I) are found in the following synthetic routes.

Example 1:

n- (4- (7H-pyrrolo [2,3-d ] pyrimidin-4-yl) benzyl) -2-chlorobenzamide

4-bromo-7-tosyl-7H-pyrrolo [2,3-d ] pyrimidine: to a 100mL round-bottomed flask was added 4-bromo-7H-pyrrolo [2,3-d ] pyrimidine (2g) and p-toluenesulfonyl chloride (2.3g), followed by acetone (50mL), water (1mL), and sodium hydroxide (0.53 g). The reaction system is reacted for 10 hours at room temperature. After the reaction is finished, the solvent is evaporated to dryness under reduced pressure, 100mL of water is added into a bottle, a white solid is separated out, the white solid is filtered, rinsed with water and a small amount of petroleum ether, and the white solid is obtained after drying. MS (ESI) M/z (M +1) +: 351.97.

(4- (7-tosyl-7H-pyrrolo [2,3-d ] pyrimidin-4-yl) phenyl) methylamine: after 4-bromo-7-tosyl-7H-pyrrolo [2,3-d ] pyrimidine (1.0g) was added to a round-bottom flask, dioxane (40mL), water (10mL), (4- (4,4,5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) phenyl) methylamine (1.0g), tetrakistriphenylphosphine palladium (0.33g), and potassium carbonate (0.59g) were added. The reaction system is heated to 100 ℃ under the protection of argon and reacted for 14 h. After the reaction was completed, the solvent was evaporated under reduced pressure, and the obtained product was dissolved in ethyl acetate. The organic phase was washed with water and saturated brine, and then dried over anhydrous sodium sulfate. Filtering the organic phase, and evaporating to dryness under reduced pressure to obtain a crude product. Purifying the crude product by pressurized silica gel column chromatography to obtain a pure product, wherein MS (ESI) M/z (M +1) +: 379.12.

2-chloro-N- (4- (7-tosyl-7H-pyrrolo [2,3-d ] pyrimidin-4-yl) benzyl) benzamide: a25 mL round bottom flask was charged with 4-bromo-7-tosyl-7H-pyrrolo [2,3-d ] pyrimidine (0.1g), 2-chlorobenzoyl chloride (0.04mL), tetrahydrofuran (6mL), and triethylamine (0.06 mL). The reaction system is reacted for 5 hours at room temperature. After the reaction, the solvent was evaporated to dryness under reduced pressure, the product was dissolved in ethyl acetate, and the organic phase was washed with water and saturated brine, and then dried over anhydrous sodium sulfate. Filtering the organic phase, and evaporating to dryness under reduced pressure to obtain a crude product. Purifying the crude product by pressurized silica gel column chromatography to obtain a pure product, wherein MS (ESI) M/z (M +1) +: 517.10.

n- (4- (7H-pyrrolo [2,3-d ] pyrimidin-4-yl) benzyl) -2-chlorobenzamide: to a 25mL round bottom flask was added 2-chloro-N- (4- (7-tosyl-7H-pyrrolo [2,3-d ] pyrimidin-4-yl) benzyl) benzamide (0.08g), followed by sodium hydroxide (0.06g), tetrahydrofuran (2mL), and water (2 mL). The reaction was stirred at room temperature for 8 h. After the reaction, the solvent was evaporated to dryness under reduced pressure, the product was dissolved in ethyl acetate, and the organic phase was washed with water and saturated brine, and then dried over anhydrous sodium sulfate. Filtering the organic phase, and evaporating to dryness under reduced pressure to obtain a crude product. Purifying the crude product by pressurized silica gel column chromatography to obtain a pure product, wherein MS (ESI) M/z (M +1) +: 363.09.

example 2:

n- (4- (7H-pyrrolo [2, 3-d)]Pyrimidin-4-yl) benzyl) -4-methylbenzamide

The synthesis of example 2 was accomplished by using a procedure similar to that described in example 1. MS (ESI) M/z (M +1) +: 343.15.

example 3:

n- (4- (1H-pyrazolo [3,4-d ] pyrimidin-4-yl) phenyl) -4-methylbenzamide

The synthesis of example 3 was accomplished by using a procedure similar to that described in example 1. MS (ESI) M/z (M +1) +: 330.13.

example 4:

n- (4- (7H-pyrrolo [2, 3-d)]Pyrimidin-4-yl) phenyl) cyclopropanecarboxamides

The synthesis of example 4 was accomplished by using a procedure similar to that described in example 1. MS (ESI) M/z (M +1) +: 279.12.

example 5:

n- (4- (7H-pyrrolo [2, 3-d)]Pyrimidin-4-yl) phenyl) -4-methylbenzamide

The synthesis of example 5 was accomplished by using a procedure similar to that described in example 1. MS (ESI) M/z (M +1) +: 329.13.

example 6:

n- (4- (7H-pyrrolo [2,3-d ] pyrimidin-4-yl) benzyl) -4-ethylbenzamide

The synthesis of example 6 was accomplished by using a procedure similar to that described in example 1. MS (ESI) M/z (M +1) +: 357.16.

example 7:

n- (4- (7H-pyrrolo [2,3-d ] pyrimidin-4-yl) benzyl) -4-isopropylbenzamide

The synthesis of example 7 was accomplished by using a procedure similar to that described in example 1. MS (ESI) M/z (M +1) +: 371.18.

example 8:

n- (4- (1H-indazol-4-yl) phenyl) -4- (tert-butyl) benzamide

The synthesis of example 8 was accomplished by using a procedure similar to that described in example 1. MS (ESI) M/z (M +1) +: 370.18.

example 9:

n- (4- (1H-indazol-4-yl) phenyl) -4-methylbenzamide

The synthesis of example 9 was accomplished by using a procedure similar to that described in example 1. MS (ESI) M/z (M +1) +: 328.14.

example 10:

n- (4- (1H-pyrrolo [2,3-b ] pyridin-4-yl) phenyl) -4- (tert-butyl) benzamide

The synthesis of example 10 was accomplished by using a procedure similar to that described in example 1. MS (ESI) M/z (M +1) +: 370.18.

example 11:

4- (7H-pyrrolo [2,3-d ] pyrimidin-4-yl) -N- (p-tolyl) benzamide

The synthesis of example 11 was accomplished by using a procedure similar to that described in example 1. MS (ESI) M/z (M +1) +: 329.38.

example 12:

4- (7H-pyrrolo [2,3-d ] pyrimidin-4-yl) -N- (4- (trifluoromethyl) phenyl) benzamide

The synthesis of example 12 was accomplished by using a procedure similar to that described in example 1. MS (ESI) M/z (M +1) +: 383.35.

example 13:

n- (4- (7H-pyrrolo [2,3-d ] pyrimidin-4-yl) benzyl) -4-fluorobenzamide

The synthesis of example 13 was accomplished by using a procedure similar to that described in example 1. MS (ESI) M/z (M +1) +: 347.12.

example 14:

n- (4- (7H-pyrrolo [2,3-d ] pyrimidin-4-yl) benzyl) -4-methylbenzamide

The synthesis of example 14 was accomplished by using a procedure similar to that described in example 1. MS (ESI) M/z (M +1) +: 343.40.

example 15:

4- (4- ((4- (tert-butyl) benzamido) methyl) phenyl) -N-methyl-1H-pyrrolo [2,3-b ] pyridine-5-carboxamide

4-chloro-N-methyl-1-tolyl-1H-pyrrolo [2,3-b ] pyridine-5-carboxamide: 1H-pyrrolo [2,3-b ] pyridine-5-carboxylic acid (1.0g) was added to a round-bottomed flask, then bis (1H-imidazol-1-yl) methanone (1.0g) and DMF (5mL) were added thereto, and after reaction for 3 hours at room temperature, a 40% aqueous methylamine solution (1.0g) was added thereto, and the reaction system was stirred at room temperature overnight. After the reaction is finished, adding 50mL of water into the system, standing for a period of time, separating out a solid, filtering and drying to obtain a crude product. To the crude product was added p-toluenesulfonyl chloride (2.1g), acetone (40mL), water (1mL), and sodium hydroxide (0.23 g). The reaction system is reacted for 10 hours at room temperature. After the reaction is finished, the solvent is evaporated to dryness under reduced pressure, 100mL of water is added into a bottle, a white solid is separated out, the mixture is filtered, rinsed with water and a small amount of petroleum ether, and a white solid crude product is obtained after drying. Purifying the crude product by pressurized silica gel column chromatography to obtain a pure product, wherein MS (ESI) M/z (M +1) +: 364.04.

4- (tert-butyl) -N- (4- (4,4,5, 5-tetramethyl-1, 3, 2-dioxaborane-2-yl) benzyl) benzamide: to a round bottom flask was added (4- (4,4,5, 5-tetramethyl-1, 3, 2-dioxaborane-2-yl) phenyl) methylamine (1.0g), followed by anhydrous DCM (10mL), 4- (tert-butyl) benzoyl chloride (1.68mL), and triethylamine (1.4 mL). The reaction system is reacted for 5 hours at room temperature. After the reaction was completed, the solvent was evaporated under reduced pressure, and the resultant was diluted with water and extracted with ethyl acetate. The organic phase was washed with water and saturated brine, and dried over anhydrous sodium sulfate. Filtering the organic phase, and evaporating to dryness under reduced pressure to obtain a crude product. Purifying the crude product by pressurized silica gel column chromatography to obtain a pure product, wherein MS (ESI) M/z (M +1) +: 394.25.

4- (4- ((4- (tert-butyl) benzamido) methyl) phenyl) -N-methyl-1H-pyrrolo [2,3-b ] pyridine-5-carboxamide: into a round-bottom flask were charged 4-chloro-N-methyl-1-tolyl-1H-pyrrolo [2,3-b ] pyridine-5-carboxamide (0.1g), 4- (tert-butyl) -N- (4- (4,4,5, 5-tetramethyl-1, 3, 2-dioxaborane-2-yl) benzyl) benzamide (0.11g), potassium carbonate (0.57g), and Pd (dppf) Cl2(0.02g), potassium iodide (0.004g), and DMF 2 mL. The reaction system is heated to 110 ℃ under the protection of argon and reacted for 14 h. After the reaction, the solvent was evaporated under reduced pressure, the obtained product was diluted with water, the aqueous phase was extracted with ethyl acetate, and the organic phase was washed with water and saturated brine, respectively, and then dried over anhydrous sodium sulfate. Filtering the organic phase, and evaporating to dryness under reduced pressure to obtain a crude product. To the crude product was added sodium hydroxide (0.11g), tetrahydrofuran (2mL), water (2 mL). The reaction was stirred at room temperature for 8 h. After the reaction, the solvent was evaporated under reduced pressure, the product was dissolved in ethyl acetate, and the organic phase was washed with water and saturated brine, respectively, and then dried over anhydrous sodium sulfate. Filtering the organic phase, and evaporating to dryness under reduced pressure to obtain a crude product. Purifying the crude product by pressurized silica gel column chromatography to obtain a pure product, wherein MS (ESI) M/z (M +1) +: 441.22.

example 16:

4- (4- (4- (tert-butyl) benzamido) phenyl) -N-methyl-1H-pyrrolo [2,3-b ] pyridine-5-carboxamide

The synthesis of example 16 was accomplished by using a procedure similar to that described in example 15. MS (ESI) M/z (M +1) +: 427.52.

example 17:

n-methyl-4- (4- (4-methyl-benzamido) phenyl) -1H-pyrrolo [2,3-b ] pyridine-5-carboxamide

The synthesis of example 17 was accomplished by using a procedure similar to that described in example 15. MS (ESI) M/z (M +1) +: 385.44.

example 18:

4- (4- (3-chlorobenzoylamino) phenyl) -N-methyl-1H-pyrrolo [2,3-b ] pyridine-5-carboxamide

The synthesis of example 18 was accomplished by using a procedure similar to that described in example 15. MS (ESI) M/z (M +1) +: 405.85.

example 19:

4- (4- (2-chlorobenzoylamino) phenyl) -N-methyl-1H-pyrrolo [2,3-b ] pyridine-5-carboxamide

The synthesis of example 19 was accomplished by using a procedure similar to that described in example 15. MS (ESI) M/z (M +1) +: 405.85.

example 20:

4- ((3- (5-fluoropyrimidin-2-yl) -2-methoxyphenyl) amino) -N-methyl-1H-pyrrolo [2,3-b ] pyridine-5-carboxamide

4-chloro-N-methyl-1-tolyl-1H-pyrrolo [2,3-b ] pyridine-5-carboxamide: 1H-pyrrolo [2,3-b ] pyridine-5-carboxylic acid (1.0g) was added to a round-bottomed flask, then bis (1H-imidazol-1-yl) methanone (1.0g) and DMF (5mL) were added thereto, and after reaction for 3 hours at room temperature, a 40% aqueous methylamine solution (1.0g) was added thereto, and the reaction system was stirred at room temperature overnight. After the reaction is finished, adding 50mL of water into the system, standing for a period of time, separating out a solid, filtering and drying to obtain a crude product. To the crude product was added p-toluenesulfonyl chloride (2.1g), acetone (40mL), water (1mL), and sodium hydroxide (0.23 g). The reaction system is reacted for 10 hours at room temperature. After the reaction is finished, the solvent is evaporated to dryness under reduced pressure, 100mL of water is added into a bottle, a white solid is separated out, the mixture is filtered, rinsed with water and a small amount of petroleum ether, and a white solid crude product is obtained after drying. Purifying the crude product by pressurized silica gel column chromatography to obtain a pure product, wherein MS (ESI) M/z (M +1) +: 364.04.

3- (5-fluoropyrimidin-2-yl) -2-methoxyaniline: to a round-bottom flask were added 4-chloro-N-methyl-1-tolyl-1H-pyrrolo [2,3-b ] pyridine-5-carboxamide (1.08g), 3- (5-fluoropyrimidin-2-yl) -2-methoxyaniline (0.7g), tetratriphenylphosphine palladium (0.46g) and potassium carbonate (1.36g), followed by dioxane (10mL) and water (2 mL). The reaction system is heated to 80 ℃ under the protection of argon and reacted for 14 h. After the reaction was completed, the solvent was evaporated under reduced pressure, and the obtained product was dissolved in ethyl acetate. The organic phase was washed with water and saturated brine, and dried over anhydrous sodium sulfate. Filtering the organic phase, and evaporating to dryness under reduced pressure to obtain a crude product. Purifying the crude product by pressurized silica gel column chromatography to obtain a pure product, wherein MS (ESI) M/z (M +1) +: 220.08.

4- ((3- (5-fluoropyrimidin-2-yl) -2-methoxyphenyl) amino) -N-methyl-1H-pyrrolo [2,3-b ] pyridine-5-carboxamide: to the dried flask was added 4-chloro-N-methyl-1-tolyl-1H-pyrrolo [2,3-b ] pyridine-5-carboxamide (0.06g), 3- (5-fluoropyrimidin-2-yl) -2-methoxyaniline (0.11g), anhydrous THF (2mL), lithium bis (trimethylsilyl) amide (0.7mL), and the reaction was reacted at room temperature under argon atmosphere for 6H. After the reaction was completed, the reaction system was quenched with a saturated aqueous ammonium chloride solution, and the resultant was neutralized with saturated sodium bicarbonate and extracted with ethyl acetate. The organic phase was washed with water and saturated brine, and dried over anhydrous sodium sulfate. Filtering the organic phase, and evaporating to dryness under reduced pressure to obtain a crude product. To the crude product was added sodium hydroxide (0.11g), tetrahydrofuran (2mL), water (2 mL). The reaction was stirred at room temperature for 8 h. After the reaction, the solvent was evaporated under reduced pressure, the product was dissolved in ethyl acetate, and the organic phase was washed with water and saturated brine, respectively, and then dried over anhydrous sodium sulfate. Filtering the organic phase, and evaporating to dryness under reduced pressure to obtain a crude product. Purifying the crude product by pressurized silica gel column chromatography to obtain a pure product, wherein MS (ESI) M/z (M +1) +: 393.14.

example 21:

effect on cancer cell proliferation

The inhibitory effect of the compounds herein on the proliferation of JAK1, JAK2, JAK3and TYK2 highly expressed cells was further evaluated by testing the effect of the compounds of the present invention on the growth of JAK1, JAK2, JAK3and TYK2 kinase highly expressed cells (table 2).

In the embodiment, the cell lines of mice Tel- (JAK1, JAK2, JAK3and TYK2) -BaF3 (stably expressing JAK1, JAK2, JAK3and TYK2 kinase) are selected and used by test companies, and the construction method comprises the following steps: human C-KIT, C-KIT 670I, PDGFR alpha, PDGFR beta, VEGFR2, FLT 3kinase region sequences were amplified by PCR respectively, and inserted into MSCV-Puro vectors (purchased from Clontech) with N-terminal TEL fragments and/or NPM fragments and/or TPR fragments, stably transferred into mouse BaF3 cells by the retrovirus method, and IL-3 growth factors were removed, finally obtaining cell lines dependent on JAK1, JAK2, JAK3and TYK2 transfer proteins.

In the examples, molecules at different concentrations (0.000508. mu.M, 0.00152. mu.M, 0.00457. mu.M, 0.0137. mu.M, 0.0411. mu.M, 0.123. mu.M, 0.370. mu.M, 1.11. mu.M, 3.33. mu.M, 10. mu.M in DMSO) were added to the above-mentioned cells, respectively, and incubated for 72 hours, the incubated cells were detected by a CCK-8 (purchased from Bebo Bio, Shanghai, China) cell viability detection kit (CCK-8 can be reduced to a yellow formazan product having high water solubility by dehydrogenase in living cells, the amount of formazan produced is proportional to the number of living cells), the number of living cells was quantified by an enzyme labeling instrument, and GI of each compound and a control compound was calculated₅₀(the results are shown in tables 2and 3).

The experimental results shown in table 2 indicate that the compounds of the present invention have certain inhibitory effects on mutant JAK1, JAK2, JAK3and TYK 2. In Table 2, "A" represents GI₅₀The value is less than 1 μ M (100nM), "B" means GI₅₀The value is in the range of 1 to 10 μ M, and "C" represents GI₅₀The value is higher than 10. mu.M.

TABLE 2

Example 22:

industrial applicability

The present invention provides a novel kinase inhibitor compound that can be used to reduce or inhibit JAK1, JAK2, JAK3and TYK2 kinase activity in a cell or a subject, and/or prevent or treat a condition associated with JAK1, JAK2, JAK3and TYK2 activity in a subject. Therefore, the compound can be prepared into corresponding medicaments and is suitable for industrial application.

Although the present invention has been described in detail herein, the present invention is not limited thereto, and modifications can be made by those skilled in the art based on the principle of the present invention, and thus, it is to be understood that various modifications made in accordance with the principle of the present invention are within the scope of the present invention.

Claims (10)

1. A kinase inhibitor comprising a compound of formula (I) or a pharmaceutically acceptable salt, solvate, ester, acid, metabolite or prodrug thereof,

wherein the content of the first and second substances,

x is a nitrogen atom or a carbon atom.

2. The kinase inhibitor according to claim 1, wherein Y is a nitrogen atom or a carbon atom, and Z is a direct bond or-NH-.

3. The kinase inhibitor of claim 2, wherein R₁Selected from hydrogen atoms, halogen atoms, C_1-6Alkyl or carboxamido radical, R₂Selected from the group consisting of optionally substituted by 1-3 independent R₃Phenyl substituted by radicalsPyridyl, pyrazolyl or pyrimidinyl.

4. The kinase inhibitor of claim 3, wherein R₃Independently selected from halogen, amino, C_1-6Alkyl radical, C_1-6Haloalkyl, C_1-6Alkoxy, (4-methylpiperazin-1-yl) methyl, methoxy-substituted phenyl, naphthyl, 4-pyridyl, 3-piperidyl or 4-piperidyl optionally substituted by methyl.

5. The kinase inhibitor according to any one of claims 1 to 4, wherein the compound is selected from,

6. a pharmaceutical composition comprising one or more kinase inhibitors according to any one of claims 1-4, and a pharmaceutically acceptable carrier or excipient, and optionally other therapeutic agents.

7. Use of one or more kinase inhibitors of any one of claims 1-4 in the manufacture of a medicament for inhibiting JAK1, JAK2, JAK3, TYK2 kinase activity.

8. Use of a pharmaceutical composition for the treatment of a disease, disorder or condition according to claim 6, wherein the disease, disorder or condition is selected from the following autoimmune diseases: arthritis, rheumatoid arthritis, osteoarthritis, lupus, rheumatoid arthritis, inflammatory bowel disease, psoriatic arthritis, osteoarthritis, still's disease, juvenile arthritis, diabetes, myasthenia gravis, hashimoto's thyroiditis, alder's thyroiditis, graves' disease, rheumatoid arthritis syndrome, multiple sclerosis, infectious neuronitis, acute disseminated encephalomyelitis, addison's disease, optic twin-myotic syndrome, ankylosing spondylitis, antiphospholipid antibody syndrome, aplastic anemia, autoimmune hepatitis, celiac disease, goodpasture's syndrome, idiopathic thrombocytopenic purpura, optic neuritis, scleroderma, primary biliary cirrhosis, reiter's syndrome, takayasu's arteritis, temporal arteritis, warm autoimmune hemolytic anemia, wegener's granulomatosis, inflammatory bowel disease, acute disseminated encephalomyelitis, addictitis encephalomyelitis, addictal encephalomyelitis, addictus disease, addison's disease, optic neuritis, optic neuropathy, myelitis obliqus syndrome, and other syndrome, myelitis obliqus syndrome, and other syndrome, Psoriasis, systemic alopecia, behcet's disease, chronic fatigue, familial autonomic dysfunction, endometriosis, interstitial cystitis, neuromuscular stiffness, scleroderma or vulvodynia.

9. Use of one or more kinase inhibitors as defined in any one of claims 1-4 in the manufacture of a medicament for the treatment or prevention of a disease, disorder or condition associated with JAK1, JAK2, JAK3, TYK2 activity.

10. The use of a medicament for the treatment of a disease, disorder, or condition according to claim 7, wherein the disease, disorder, or condition is a JAK1, JAK2, JAK3, TYK2 kinase-associated disease.