Janus kinase (JAK) family inhibitors and their preparation and use

Abstract

To provide 7-azaindole JAK kinase family inhibitors.SOLUTION: The present invention relates to a 7-azaindole derivative having the structure of formula (I), a pharmaceutical composition containing the compound of formula (I), and uses of the compound in preparing a medicament for preventing or treating Janus kinase (JAK) family-related diseases, specifically, uses in preventing or treating inflammatory diseases related to protein tyrosine kinase. In the formula (I), each substituent is as defined in the specifications.SELECTED DRAWING: None

Description

[Technical field]

[0001] The present application belongs to the technical field of medicine, specifically relates to 7-azaindole derivatives and their use for preparing anti-inflammatory drugs. [Background technology]

[0002] Janus kinase (JAK) is a type of tyrosine kinase, and its family includes four members: JAK1, JAK2, JAK3, and TYK2. JAKs play an important role in the transmission of various cytokine signals, and together with their downstream signal transducers and activators of transcription (STATs), mediate the regulation of gene transcription and expression in the cell nucleus by cytokine receptors.

[0003] Cytokines bind to their receptors, causing the dimerization of the receptor molecules, and the JAKs coupled to the receptors approach each other and are activated by phosphorylation of the interacting tyrosine residues. The activated JAKs catalyze the phosphorylation of the tyrosine residues of the receptor itself, forming the corresponding sites of STATs that bind to the receptor complex. The SH2 domains of STATs bind to phosphotyrosine residues of the receptor molecule, and phosphorylation of the C-terminal tyrosine residues is accomplished by the action of JAKs. Two phosphorylated STAT molecules interact with each other to form homologous / heterologous dimers, detach from the receptor molecule, enter the cell nucleus, bind to the promoter regions of target genes, and regulate gene transcription and expression (J. Biol Chem. 2007, 282, 20059).

[0004] The JAK / STAT system is the most important intracellular signal transduction system for cytokines in immune cells, and the combination of four JAKs and seven STATs transmits signals of about 40 types of cytokines (Immunol Rev. 2009, 228, 273). JAK3 can specifically bind to the common gamma chain (Fcγ) of cytokine receptors, while JAK1 binds to the beta chain, and both of them are activated by IL-2, IL-4, IL-7, IL-9, IL-15, and IL-21 cytokines (Immunol. Rev. 2009, 228, 273). Some important inflammation-correlated cytokines, such as IL-6, transmit signals only through JAK1 (EMBO J. 1995, 14, 1421). IL-6 monoclonal antibodies such as tocilizumab have been proven to be effective drugs in the treatment of rheumatoid arthritis (RA) (Arthritis Rheum. 2006, 54, 2817), so JAK1 is an ideal anti-inflammatory target. JAK2 plays an important role in the erythropoietin (EPO) signaling pathway, such as promoting erythrocyte differentiation and activating STAT5. In addition, JAK2 is also involved in the lipid metabolism pathway (Obesity. 2008, 16, 492). JAK1, JAK2 and TYK2 are widely present in various tissues and cells, while JAK3 is mainly distributed in lymphocytes, and excessive inhibition increases the risk of infection. Aberrant cytokine generation and abnormal cytokine signaling are not only related to various immune and inflammatory diseases such as autoimmune diseases and allergies, but also to diseases with various different pathologies such as cancer. The regulation of many abnormal immune responses, including allergies, asthma, (xeno)transplant rejection, autoimmune diseases such as rheumatoid arthritis, amyotrophic lateral sclerosis, and multiple sclerosis, and hematological malignancies such as myeloproliferative disorders, leukemia, and lymphoma, is associated with the JAK / STAT signaling pathway.

[0005] Tofacitinib is the first commercially available oral JAK inhibitor, but because it is a pan-JAK inhibitor, it has low selectivity and inhibits JAK1 as well as JAK2 and JAK3. This results in clinically significant side effects such as anemia and infection, which affect the clinical dose and final therapeutic effect. (Transplantation. 2005, 79, 791; Annals of the Rheumatic Diseases. 2016, 75, 1133.). Baricitinib, a JAK1 / JAK2 inhibitor, has shown superior anti-inflammatory therapeutic effects to Humira (adalimumab) in clinical trials, but has a clear side effect of increasing blood lipids (Ann Rheum Dis. 2018, 77, 988.), so a low-dose formulation approved by the FDA is available on the market. ABT-494, which selectively inhibits JAK1, has also shown superior anti-inflammatory therapeutic effects in clinical trials, but there is a clinical risk of increasing low-density lipoprotein (LDL) (Arthritis & Rheumatology. 2016, 68, 2857). Therefore, the development of JAK inhibitors with better selectivity remains of important clinical significance.

[0006] Tyk2 is also a member of the JAK kinase family and, as an important immunoregulatory site, is involved in the transmission of IFN-α, IL-6, IL-10, IL-12, and IL-23 signal pathways, and transmits inflammatory signals by phosphorylating IL-12, IL-23, and STAT proteins downstream of type I interferon receptor. Since the above inflammatory factors are involved in the development of various autoimmune diseases, such as systemic lupus erythematosus (SLE), guttate psoriasis (PSO), and inflammatory bowel disease (IBD), Tyk2 is also an important therapeutic target for inflammatory diseases. Currently, several Tyk2 inhibitors are used in clinical trial studies of systemic lupus erythematosus, guttate psoriasis, inflammatory bowel disease, and alopecia areata (J. Med. Chem. 2018, 61, 8597; J. Med. Chem., 2018, 61, 8594).

[0007] WO2018169700 and WO2016116025 disclose JAK inhibitor compounds, but the compounds of the present invention differ in structure from these compounds and have overall superior inhibitory activity against JAK1 to the compounds of the WO2018169700 and WO2016116025 patents. Summary of the Invention

[0008] An object of the present invention is to provide a 7-azaindole JAK kinase family inhibitor.

[0009] Another object of the present invention is to provide a use of a highly selective JAK kinase family inhibitor for the preparation of a drug for preventing or treating a JAK kinase family-related disease.

[0010] The present invention relates to a compound represented by the following formula (I), a stereoisomer thereof, or a pharma- ceutically acceptable salt thereof: [ka] wherein Ring A is selected from optionally substituted 4- to 12-membered heterocyclyl or 5- to 10-membered heteroaryl; R 1 is hydrogen, hydroxy, optionally substituted C 1 -C 8 Alkyl, C 3 -C 8 Cycloalkyl, C 2 -C 8 Alkenyl, C 2 -C 8 selected from alkynyl, 4- to 12-membered heterocyclyl, 6- to 10-membered aryl, or 5- to 10-membered heteroaryl; R 2 is cyano, -C=ONR 6 R 7 , -C=ONR 6 NR 7 R 8 , -C=ONHOR 6 , -S(O) m R 8 , -S(O) m -NHR 8 , or -C=OOR 6 Selected from L is amino, -NR 6 C=O-, -NR 6 C=ONR 10 -,-C=ONR 10 -,-C=ONR6 O-, -C=ONR 6 NR 10 -, -NR 6 S(O) m -, -S(O) m NR 6 -, -NR 6 S(O) m NR 7 -, -S(O) m -, -C=O- or -C=OO-, or L is absent; R 3 is hydrogen, optionally substituted C 1 -C 8 Alkyl, C 3 -C 8 Cycloalkyl or C=OR 6 Selected from R 4 and R 5 are hydrogen, deuterium, halogen, cyano, nitro, and optionally substituted C 1 -C 8 Alkyl, C 3 -C 8 independently selected from cycloalkyl, 6- to 10-membered aryl, or 5- to 10-membered heteroaryl; R 6 , R 7 and R 10 are each hydrogen, optionally substituted C 1 -C 8 Alkyl or C 3 -C 8 independently selected from cycloalkyl; Or, R 10 is the N and R bonded to it 1 together with Here, the rings A and R 1 The substituents in each of the formulas are each one or more of halogen, hydroxy, cyano, amino, mercapto, nitro, C 1 -C 8 Alkyl, C 1 -C 8 Alkoxy, C 3 -C 8Cycloalkyl, 4-12 membered heterocyclyl, 6-10 membered aryl, 5-10 membered heteroaryl, -(CH 2 ) n C=OOR 8 , -OC=OR 8 , -C=OR 8 , -C=ONR 8 R 9 , -NHC=OR 8 , -NR 8 R 9 , -OC=ONR 8 R 9 , -NHC=ONR 8 R 9 , -S(O) m R 8 , -S(O) m -NHR 8 , -NHC=OOR 8 or -NHS(O) m R 8 Independently selected from m is selected from 1 or 2, n is selected from 1, 2, 3, 4 or 5; R 8 and R 9 are each hydrogen, optionally substituted C 1 -C 8 Alkyl or C 3 -C 8 independently selected from cycloalkyl; The above R 3 , R 4 , R 5 , R 6 , R 7 , R 8 , R 9 and R 10 The substituents in each of the formulas are each one or more of deuterium, halogen, hydroxy, cyano, amino, mercapto, nitro, C 1 -C 8 Alkyl, C 1 -C 8 Alkoxy or C 3 -C 8 cycloalkyl.

[0011] In some embodiments of the present invention, Ring A is selected from optionally substituted 4-12 membered heterocyclyl, R 1 is hydrogen, hydroxy, optionally substituted C 1 -C 8 Alkyl, C 3 -C 8 Cycloalkyl, C 2 -C 8 Alkenyl, C 2 -C 8 selected from alkynyl, 4- to 12-membered heterocyclyl, 6- to 10-membered aryl, or 5- to 10-membered heteroaryl; R 2 is cyano or -C=ONR 6 R 7 Selected from L is amino, -NR 6 C=O-, -NR 6 C=ONR 10 -,-C=ONR 10 -,-C=ONR 6 O-, -C=ONR 6 NR 10 -, -NR 6 S(O) m -, -S(O) m NR 6 -, -NR 6 S(O) m NR 7 -, -S(O) m -, -C=O- or -C=OO-, or L is absent; R 3 is hydrogen, optionally substituted C 1 -C 8 Alkyl, C 3 -C 8 Cycloalkyl or -C=OR 6 Selected from R 4 and R 5 are hydrogen, deuterium, halogen, cyano, nitro, and optionally substituted C 1 -C 8 Alkyl or C 3 -C 8 independently selected from cycloalkyl; R6 , R 7 and R 10 are each hydrogen, optionally substituted C 1 -C 8 Alkyl or C 3 -C 8 independently selected from cycloalkyl; Or, R 10 is the N and R bonded to it 1 together with Here, the rings A and R 1 The substituents in each of the formulas are each one or more of halogen, hydroxy, cyano, amino, C 1 -C 8 Alkyl, C 1 -C 8 Alkoxy, C 3 -C 8 Cycloalkyl, 4-12 membered heterocyclyl, 6-10 membered aryl, 5-10 membered heteroaryl, -C=ONR 8 R 9 , -NHC=OR 8 , -NR 8 R 9 , -OC=ONR 8 R 9 , -NHC=ONR 8 R 9 , -S(O) m R 8 , -S(O) m -NHR 8 , -NHC=OOR 8 or -NHS(O) m R 8 Independently selected from m is selected from 1 or 2, R 8 and R 9 are each hydrogen, optionally substituted C 1 -C 8 Alkyl or C 3 -C 8 independently selected from cycloalkyl; The above R 3 , R 4 , R 5 , R 6, R 7 , R 8 , R 9 and R 10 The substituents in each of the formulas are each one or more of deuterium, halogen, hydroxy, cyano, amino, C 1 -C 8 Alkyl, C 1 -C 8 Alkoxy or C 3 -C 8 cycloalkyl. In some embodiments of the present invention, Ring A is selected from optionally substituted 4-12 membered heterocyclyl, R 1 is hydrogen, hydroxy, optionally substituted C 1 -C 8 Alkyl, C 3 -C 8 Cycloalkyl, C 2 -C 8 Alkenyl, C 2 -C 8 selected from alkynyl, 4- to 12-membered heterocyclyl, 6- to 10-membered aryl, or 5- to 10-membered heteroaryl; R 2 is cyano or -C=ONR 6 R 7 Selected from L is amino, -NR 6 C=O-, -NR 6 C=ONR 10 -,-C=ONR 10 -,-C=ONR 6 O-, -C=ONR 6 NR 10 -, -NR 6 S(O) m -, -S(O) m NR 6 -, -NR 6 S(O) m NR 7 -, -S(O) m -, -C=O- or -C=OO-, or L is absent; R 3 is selected from hydrogen or methyl; R 4 and R5 are hydrogen, deuterium, halogen, cyano, nitro, and optionally substituted C 1 -C 8 Alkyl or C 3 -C 8 independently selected from cycloalkyl; R 6 , R 7 and R 10 are each hydrogen, optionally substituted C 1 -C 8 Alkyl or C 3 -C 8 independently selected from cycloalkyl; Or, R 10 is the N and R bonded to it 1 together with Here, the rings A and R 1 The substituents in each of the formulas are each one or more of halogen, hydroxy, cyano, amino, C 1 -C 8 Alkyl, C 1 -C 8 Alkoxy, C 3 -C 8 Cycloalkyl, 4-12 membered heterocyclyl, 6-10 membered aryl, 5-10 membered heteroaryl, -C=ONR 8 R 9 , -NHC=OR 8 , -NR 8 R 9 , -OC=ONR 8 R 9 , -NHC=ONR 8 R 9 , -S(O) m R 8 , -S(O) m -NHR 8 , -NHC=OOR 8 or -NHS(O) m R 8 Independently selected from m is selected from 1 or 2, R 8 and R 9 are each hydrogen, optionally substituted C 1-C 8 Alkyl or C 3 -C 8 independently selected from cycloalkyl; The above R 4 , R 5 , R 6 , R 7 , R 8 , R 9 and R 10 The substituents in each of the formulas are each one or more of deuterium, halogen, hydroxy, cyano, amino, C 1 -C 8 Alkyl, C 1 -C 8 Alkoxy or C 3 -C 8 cycloalkyl.

[0012] In some embodiments of the present invention, Ring A is selected from the following optionally substituted groups: [ka] R 1 is hydrogen, hydroxy, optionally substituted C 1 -C 8 Alkyl, C 3 -C 8 selected from cycloalkyl, 4- to 12-membered heterocyclyl, 6- to 10-membered aryl, or 5- to 10-membered heteroaryl; R 2 is cyano or -C=ONR 6 R 7 Selected from L is amino, -NR 6 C=O-, -NR 6 C=ONR 10 -,-C=ONR 10 -,-C=ONR 6 O-, -C=ONR 6 NR 10 -, -NR 6 S(O) m -, -S(O) m NR 6 -, -NR 6 S(O) m NR7 -, -S(O) m -, -C=O- or -C=OO-; Or, L is not present, R 3 is selected from hydrogen or methyl; R 4 and R 5 are hydrogen, deuterium, halogen, cyano, nitro, and optionally substituted C 1 -C 8 Alkyl or C 3 -C 8 independently selected from cycloalkyl; R 6 , R 7 and R 10 are each hydrogen, optionally substituted C 1 -C 8 Alkyl or C 3 -C 8 independently selected from cycloalkyl; Or, R 10 is the N and R bonded to it 1 together with m is selected from 1 or 2, Here, the rings A and R 1 The substituents in each of the formulas are each one or more of halogen, hydroxy, cyano, amino, C 1 -C 8 Alkyl, C 1 -C 8 Alkoxy, C 3 -C 8 Cycloalkyl, 4-12 membered heterocyclyl, 6-10 membered aryl, 5-10 membered heteroaryl, -C=ONR 8 R 9 , -NHC=OR 8 , -NR 8 R 9 , -OC=ONR 8 R 9 , -NHC=ONR 8 R 9 , -S(O) m R 8 , -S(O) m -NHR8 , -NHC=OOR 8 or -NHS(O) m R 8 Independently selected from R 8 and R 9 are each hydrogen, optionally substituted C 1 -C 8 Alkyl or C 3 -C 8 independently selected from cycloalkyl; The above R 4 , R 5 , R 6 , R 7 , R 8 , R 9 and R 10 The substituents in each of the formulas are each one or more of deuterium, halogen, hydroxy, cyano, amino, C 1 -C 8 Alkyl, C 1 -C 8 Alkoxy or C 3 -C 8 cycloalkyl.

[0013] In some embodiments of the present invention, Ring A is selected from the following optionally substituted groups: [ka] R 1 is hydrogen, hydroxy, optionally substituted methyl, ethyl, propyl, cyclopropyl, n-butyl, t-butyl, cyclobutyl, phenyl, pyridinyl, imidazolyl, pyrazolyl, oxazolyl, thiazolyl or [ka] Selected from R 2 is cyano or -C=ONR 6 R 7 Selected from L is amino, -NR 6 C=O-, -NR 6C=ONR 10 -,-C=ONR 10 -,-C=ONR 6 O-, -C=ONR 6 NR 10 -, -NR 6 S(O) m -, -S(O) m NR 6 -, -NR 6 S(O) m NR 7 -, -S(O) m -, -C=O- or -C=OO-; Or, L is not present, R 3 is selected from hydrogen or methyl; R 4 and R 5 are each independently selected from hydrogen, deuterium, halogen, or cyano; R 6 , R 7 and R 10 are each hydrogen, optionally substituted C 1 -C 8 Alkyl or C 3 -C 8 independently selected from cycloalkyl; Or, R 10 is the N and R bonded to it 1 together with m is selected from 1 or 2, Here, the rings A and R 1 The substituents in each of the formulas are each one or more of halogen, hydroxy, cyano, amino, C 1 -C 8 Alkyl, C 1 -C 8 Alkoxy, C 3 -C 8 Cycloalkyl, 4-12 membered heterocyclyl, 6-10 membered aryl or 5-10 membered independently selected from heteroaryl; The above R 6 , R 7 and R 10The substituents in each of the formulas are each one or more of deuterium, halogen, hydroxy, cyano, amino, C 1 -C 8 Alkyl, C 1 -C 8 Alkoxy or C 3 -C 8 cycloalkyl.

[0014] In some embodiments of the present invention, Ring A is selected from the following optionally substituted groups: [ka] R 1 is hydrogen, hydroxy, optionally substituted methyl, ethyl, propyl, cyclopropyl, n-butyl, t-butyl, cyclobutyl, phenyl, pyridinyl, imidazolyl, pyrazolyl, oxazolyl, thiazolyl or [ka] Selected from R 2 is cyano or -C=ONR 6 R 7 Selected from L is amino, -NR 6 C=O-, -C=ONR 10 -, -S(O) m -, -C=O- or -C=OO-; Or, L is not present, R 3 is selected from hydrogen or methyl; R 4 and R 5 are each independently selected from hydrogen, deuterium, fluorine, chlorine, or cyano; R 6 , R 7 and R 10 are each hydrogen, optionally substituted C 1 -C 8 Alkyl or C 3 -C 8independently selected from cycloalkyl; Or, R 10 is the N and R bonded to it 1 together with m is selected from 1 or 2, Here, the rings A and R 1 The substituents in each of the formulas are each one or more of halogen, hydroxy, cyano, amino, C 1 -C 4 Alkyl or C 1 -C 4 independently selected from alkoxy, The above R 6 , R 7 and R 10 Each substituent in is independently selected from one or more deuterium, halogen, hydroxy, cyano, or amino.

[0015] In some embodiments of the present invention, Ring A is selected from the following optionally substituted groups: [ka] R 1 is hydrogen, hydroxy, optionally substituted methyl, ethyl, propyl, cyclopropyl, t-butyl, cyclobutyl or [ka] Selected from R 2 is cyano or -C=ONR 6 R 7 Selected from R 6 and R 7 is selected from hydrogen, methyl or deuterated methyl; L is -C=ONR 10 -, -S(O) m -, -C=O- or -C=OO-; Or, L is not present, R 3is selected from hydrogen or methyl; R 4 and R 5 are each independently selected from hydrogen or deuterium; R 10 is hydrogen, m is selected from 1 or 2, Here, the rings A and R 1 Each substituent in is independently selected from one or more of fluorine, chlorine, hydroxy, cyano, amino, methyl or methoxy.

[0016] In some embodiments of the present invention, the compound is the compound shown below or a pharma- ceutically acceptable salt thereof.

[0017] [ka]

[0018] [ka]

[0019] Preferably, the compound of the present invention is the compound shown below or a pharma- ceutically acceptable salt thereof.

[0020] [ka]

[0021] [ka]

[0022] Another aspect of the present invention provides a process for preparing a compound of formula (I), a stereoisomer thereof or a pharma- ceutically acceptable salt thereof, which comprises the steps of:

[0023] [ka] Here, A, L, R 1 , R 2 , R 3 , R 4 and R 5 is as defined above for compounds of formula (I).

[0024] The present invention further provides a pharmaceutical composition comprising a compound of formula (I), a stereoisomer or a pharma- ceutically acceptable salt thereof, and a pharma- ceutical acceptable carrier.

[0025] Preferably, the pharmaceutical composition is selected from capsules, powders, tablets, pellets, pills, injections, syrups, oral liquids, inhalants, ointments, suppositories or patches.

[0026] Another aspect of the present invention provides the use of a compound of formula (I) or its tautomers, mesomers, racemates, enantiomers, diastereoisomers, and mixtures thereof, and pharma- ceutically acceptable salts thereof, or a pharmaceutical composition containing the same, for the preparation of a medicament for preventing or treating a Janus kinase (JAK) family-induced disease, including immune system diseases, autoimmune diseases, skin diseases, allergic diseases, viral diseases, type 1 diabetes and diabetic complications, Alzheimer's disease, xerophthalmia, myelofibrosis, thrombocytosis, erythrocytosis, leukemia, and cancer, the immune system diseases being organ transplant rejection, xenograft rejection, or graft-versus-host disease, the autoimmune diseases being selected from systemic lupus erythematosus, multiple sclerosis, rheumatoid arthritis, juvenile arthritis, guttate psoriasis, ulcerative colitis, Crohn's disease, and autoimmune thyroid disease, and the skin diseases being psoriasis, skin rash, alopecia areata, and the like. or atopic dermatitis, the allergic disease is selected from asthma or rhinitis, the viral disease is selected from hepatitis B, hepatitis C, chicken pox, and varicella zoster virus, the cancer is selected from solid tumors, blood cancer, or skin cancer, the solid tumor is selected from prostate cancer, kidney cancer, liver cancer, pancreatic cancer, gastric cancer, breast cancer, lung cancer, head and neck cancer, thyroid cancer, spongioblastoma, or melanoma, the blood cancer is selected from lymphoma or leukemia, and the skin cancer is selected from cutaneous T-cell lymphoma or cutaneous B-cell lymphoma.

[0027] Unless otherwise stated, terms used in the specification and claims have the following meanings.

[0028] In the present invention, "aryl" refers to an all-carbon monocyclic or bicyclic group, and "6-10 membered aryl" refers to an all-carbon aryl having 6-10 carbons, e.g., phenyl, naphthyl. The aryl ring may be fused to a heteroaryl, heterocyclyl, or cycloalkyl ring, where the ring attached to the parent structure is the aryl ring.

[0029] In the present invention, the term "heteroaryl" refers to a heteroaryl system having 1 to 4 heteroatoms, including nitrogen, oxygen, and sulfur heteroatoms, such as furyl, thienyl, pyridinyl, pyrrolyl, pyrazolyl, N-alkylpyrrolyl, pyrimidinyl, pyrazinyl, imidazolyl, tetrazolyl, oxazolyl, or the like. [ka] etc.

[0030] In the present invention, "C 1 -C 8 "Alkyl" refers to straight chain and branched chain alkyl groups having 1 to 8 carbon atoms, where alkyl refers to saturated aliphatic hydrocarbon groups, such as methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl, t-butyl, s-butyl, n-pentyl, 1,1-dimethylpropyl, 1,2-dimethylpropyl, 2,2-diphenylpropyl, 2,3-diphenylpropyl, 2,4-diphenylpropyl, 2,5-diphenylpropyl, 2,6-diphenylpropyl, 2,7-diphenylpropyl, 2,8-diphenylpropyl, 2,9-diphenylpropyl, 3,10-diphenylpropyl, 3,11-diphenylpropyl, 3,12-diphenylpropyl, 3,13-diphenylpropyl, 3,14-diphenylpropyl, 3,15-diphenylpropyl, 3,16-diphenylpropyl, 3,17-diphenylpropyl, 3,18-diphenylpropyl, 3 Methylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, 1-ethyl-2-methylpropyl, 1,1,2-trimethylpropyl, 1,1-dimethylbutyl, 1,2-dimethylbutyl, 2,2-dimethylbutyl, 1,3-dimethylbutyl, 2-ethylbutyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, 2,3-dimethylbutyl, n-heptyl, 2-methylhexyl, 3-methylhexyl, 4-methylhexyl, 5-methylhexyl, 2,3-dimethylpentyl and the like. Examples of the aryl groups include 2,4-dimethylpentyl, 2,2-dimethylpentyl, 3,3-dimethylpentyl, 2-ethylpentyl, 3-ethylpentyl, n-octyl, 2,3-dimethylhexyl, 2,4-dimethylhexyl, 2,5-dimethylhexyl, 2,2-dimethylhexyl, 3,3-dimethylhexyl, 4,4-dimethylhexyl, 2-ethylhexyl, 3-ethylhexyl, 4-ethylhexyl, 2-methyl-2-ethylpentyl, 2-methyl-3-ethylpentyl, and branched chain isomers thereof.

[0031] As used herein, "cycloalkyl" refers to a saturated monocyclic hydrocarbon substituent, including "C 3 -C 8 "Cycloalkyl" refers to a monocyclic cycloalkyl having 3 to 8 carbon atoms, examples of monocyclic cycloalkyl include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, and the like.

[0032] In the present invention, "alkenyl" refers to an alkyl as defined above having at least two carbon atoms and at least one carbon-carbon double bond, and "C 2 -C 8 "Alkenyl" refers to an alkenyl having 2 to 8 carbons, including straight or branched chains, such as ethenyl, 1-propenyl, 2-propenyl, 1-butenyl, 2-butenyl, or 3-butenyl.

[0033] As used herein, "alkynyl" refers to an alkyl group as defined above having at least two carbon atoms and at least one carbon-carbon triple bond, and includes "C 2 -C 8 "Alkynyl" refers to an alkynyl group containing straight or branched chains having 2 to 8 carbons, such as ethynyl, 1-propynyl, 2-propynyl, 1-butynyl, 2-butynyl, or 3-butynyl.

[0034] If a substituent bond "---" is attached to a particular position, the substituent is substituted at this position; If a substituent bond "---" is a cross-connect between two atoms on the same ring, then the substituent can be bonded to any atom on the ring, i.e., the substituent can be substituted at any position on the ring. For example, the structural unit [ka] can be substituted at any one position on the cyclohexyl.

[0035] As used herein, "heterocyclyl" refers to a saturated or partially unsaturated monocyclic or polycyclic cyclic hydrocarbon substituent in which one or more ring atoms are nitrogen, oxygen or S(O) m and the remaining ring atoms, excluding -OO-, -OS-, or -SS- in the ring, are carbon. "4-12 membered heterocyclyl" refers to a ring group having 4 to 12 ring atoms. Examples of monocyclic heterocyclyl include, but are not limited to, pyrrolidinyl, piperidinyl, piperazinyl, morpholinyl, thiomorpholinyl, higher piperazinyl, and the like, and polycyclic heterocyclyl includes spiro, fused, and bridged heterocyclyl, including, but not limited to, the following structures:

[0036] [ka]

[0037] As used herein, "alkoxy" refers to -O-(alkyl), where alkyl is as defined above. 1 -C 8 "Alkoxy" refers to alkyloxy having 1 to 8 carbons, examples of which include, but are not limited to, methoxy, ethoxy, propoxy, butoxy, and the like.

[0038] "Halogen" refers to fluorine, chlorine, bromine or iodine.

[0039] The term "pharmaceutical composition" refers to a mixture containing one or more of the above-mentioned compounds or their physiologically / pharmacologically acceptable salts or prodrugs together with other chemical components, such as physiologically / pharmacologically acceptable carriers and excipients. The purpose of a pharmaceutical composition is to facilitate administration to a living body, thereby contributing to the absorption of the active ingredient and exhibiting biological activity.

[0040] A "carrier" refers to a material that is not significantly irritating to the living body and does not abrogate the biological activity and properties of the compound being administered.

[0041] The abbreviations of the reagents used in the preparation process of the present invention are as follows. DMF N,N-Dimethylformamide THF Tetrahydrofuran PE Petroleum Ether EA Ethyl acetate CDI Carbonyldiimidazole TBTU O-Benzotriazole-N,N,N',N'-tetramethyluronium tetrafluoroborate DCM Dichloromethane Boc t-butoxycarbonyl TIPSCl Triisopropylsilyl Chloride MeCN Acetonitrile TFA Trifluoroacetic acid [Brief description of the drawings]

[0042] [Figure 1]1 is a 1H NMR spectrum of the compound of Example 1. [Diagram 2] 1H NMR spectrum of the compound of Example 2. [Diagram 3] 1H NMR spectrum of the compound of Example 3. [Figure 4] 1H NMR spectrum of the compound of Example 4. [Diagram 5] 1H NMR spectrum of the compound of Example 5. [Figure 6] 1H NMR spectrum of the compound of Example 6. [Figure 7] 1H NMR spectrum of the compound of Example 7. [Figure 8] 1H NMR spectrum of the compound of Example 8. [Figure 9] 1H NMR spectrum of the compound of Example 9. [Figure 10] 1H NMR spectrum of the compound of Example 10. [Figure 11] 1H NMR spectrum of the compound of Example 11. [Figure 12] 1H NMR spectrum of the compound of Example 12. [Figure 13] 1H NMR spectrum of the compound of Example 13. [Figure 14] 1H NMR spectrum of the compound of Example 14. [Figure 15] 1H NMR spectrum of the compound of Example 15. [Figure 16] 1H NMR spectrum of the compound of Example 16. [Figure 17] 1H NMR spectrum of the compound of Example 17. [Figure 18] 1H NMR spectrum of the compound of Example 18. [Figure 19] 1H NMR spectrum of the compound of Example 19. [Figure 20] 1H NMR spectrum of the compound of Example 20. [Figure 21] 1H NMR spectrum of the compound of Example 21. [Figure 22] 1H NMR spectrum of the compound of Example 22. [Figure 23] 1H NMR spectrum of the compound of Example 23. [Figure 24] This shows the results of evaluation of disease activity index (DAI) in a mouse DSS colitis model. [Diagram 25] FIG. 1 shows the effect of colon weight to colon length ratio in a rat DNBS-induced Crohn's disease model. [Figure 26] 1 shows the area of ​​colonic ulcers in a rat DNBS-induced Crohn's disease model. [Figure 27] Photographs of the colon of G1 rats. [Figure 28] Photographs of the colon of G2 rats. [Figure 29] Photographs of the colon of G3 rats. [Diagram 30] Photographs of the colon of G4 rats. [Diagram 31] Photographs of the colon of G5 rats. [Diagram 32] Photographs of the colon of G6 rats. [Diagram 33] The photographs show reduced colons of rats from groups G1 to G6 for comparison. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0043] The present invention will now be described with reference to specific examples. Those skilled in the art can understand that these examples are used to illustrate the present invention and do not limit the scope of the present invention in any way.

[0044] In the following examples, the experimental methods are conventional unless otherwise specified. The raw materials and reagents used in the following examples are commercially available products unless otherwise specified.

[0045] Example 1 Synthesis of (S)-4-(3-(2,2,2-trifluoroethylcarbamoyl)piperidine-1-amino)-1H-7-azaindole-5-formamide [ka]

[0046] Step 1: Synthesis of 4-chloro-1-(triisopropylsilyl)-7-azaindole [ka]

[0047] At room temperature, 4-chloro-7-azaindole (100.00 g, 655.39 mmol) was dissolved in DMF (1.2 L), cooled to 0 ° C in an ice bath, NaH (39.47 g, 983.09 mmol) was added in portions, and then stirred at 0 ° C for 1 hour. Triisopropylsilyl chloride (190.80 g, 983.09 mmol, abbreviation TIPSCl) was added dropwise, and the mixture was warmed to room temperature and reacted for 2 hours. The reaction solution was poured into 2 L of ice water, extracted with petroleum ether (1 L x 2), the organic phase was combined, washed with saturated saline (1 L x 3), concentrated to dryness under reduced pressure, and 197.50 g of a colorless transparent liquid was obtained by column chromatography (PE:EA = 1:0) in a yield of 97%.

[0048] Step 2: Synthesis of ethyl 4-chloro-1-(triisopropylsilyl)-7-azaindole-5-carboxylate [ka]

[0049] In a four-neck flask, 4-chloro-1-(triisopropylsilyl)-7-azaindole (20.00 g, 64.74 mmol) was dissolved in THF (100 mL), cooled to -75 ° C, s-butyl lithium (100 mL, 129.48 mmol, Sec. BuLi) was added dropwise, and the mixture was stirred at -75 ° C for 1 hour. Chloroethyl chlorocarboxylate (17.80 g, 129.48 mmol) was added, and the mixture was reacted at -75 ° C for 1 hour. The reaction liquid was poured into a saturated ammonium chloride solution, separated, and the aqueous phase was extracted with ethyl acetate (100 mL). The organic phases were combined, washed with saturated saline, dried over anhydrous sodium sulfate, filtered, concentrated, and column (PE:EA = 1:0-10:1) to obtain 23.15 g of a pale yellow liquid in a yield of 94%.

[0050] Step 3: Synthesis of 4-chloro-7-azaindole-5-carboxylic acid [ka]

[0051] Ethyl 4-chloro-1-(triisopropylsilyl)-7-azaindole-5-carboxylate (12.94 g, 33.96 mmol) was added to a single-neck flask at room temperature, and then ethanol (200 mL) and 10% sodium hydroxide (100 mL) were added, followed by reaction at 60° C. for 2 hours. The ethanol was removed under reduced pressure, and the aqueous phase was adjusted to pH=4 with 1N dilute hydrochloric acid in an ice bath. The white solid that was eluted in large quantities was filtered, and the filter cake was dried to obtain 8.60 g of a white solid in a yield of 77%.

[0052] Step 4: Synthesis of 4-chloro-7-azaindole-5-formamide [ka]

[0053] In a four-neck flask, 4-chloro-7-azaindole-5-carboxylic acid (7.0 0g, 35.60mmol) was added, 100mL of DMF was added, carbonyldiimidazole (8.72g, 53.41mmol, CDI) was added while stirring, and the mixture was stirred at room temperature for 1.5 hours, ammonia water (9.34g, 142.40mmol) was added dropwise at 0°C, the mixture was allowed to warm to room temperature naturally, and the mixture was stirred for 2 hours to react. 100mL of EA was added to the reaction solution, and the white solid that was dissolved in large quantities by leaving it to stand was filtered, and the filter cake was washed and dried to obtain 5.34g of a gray solid in a yield of 76%.

[0054] Step 5: Synthesis of (S)-1-nitrosopiperidine-3-carboxylate ethyl [ka]

[0055] At room temperature, (S)-ethyl 3-piperidinecarboxylate (10.00 g, 0.064 mol) was added to a mixed solution consisting of glacial acetic acid (100 mL) and water (40 mL), and the mixture was cooled to 0°C, and then 20 mL of an aqueous solution containing dissolved sodium nitrite (8.78 g, 0.13 mol) was added dropwise, and the mixture was reacted for 1 hour while stirring at 0°C, and then the mixture was heated to room temperature and reacted for 2 hours. 200 mL of water was added to the reaction solution, and the mixture was extracted with ethyl acetate (200 mL x 3). The organic phases were combined, washed with saturated saline, concentrated under reduced pressure, and then directly introduced into the next step.

[0056] Step 6: Synthesis of (S)-1-aminopiperidine-3-carboxylate ethyl salt [ka]

[0057] At room temperature, (S)-ethyl 1-nitrosopiperidine-3-carboxylate (11.83 g, 0.064 mol) was dissolved in methanol (100 mL), Zn powder (10.40 g, 0.16 mol) was added, the temperature was lowered to -5°C, glacial acetic acid (50 mL) was slowly added dropwise, and the mixture was reacted for 0.5 hours with stirring at 0°C, and the mixture was warmed to room temperature and reacted for 2 hours. The reaction solution was filtered, the filter cake was poured into 100 mL of methanol, the filtrate was evaporated to dryness, an ethanol solution of hydrochloric acid was added to the residue, the mixture was stirred for 0.5 hours, and the solvent was distilled off to obtain 13.27 g of a yellow oily product in a 100% yield of the crude product of steps 5 and 6.

[0058] Step 7: Synthesis of ethyl (S)-1-(5-formylamino-1H-7-azaindole-4-amino)piperidine-3-carboxylate [ka]

[0059] 4-Chloro-7-azaindole-5-formamide (1.00 g, 5.12 mmol) and (S)-ethyl 1-aminopiperidine-3-carboxylate hydrochloride (2.14 g, 10.24 mmol) were added to a microwave tube, 20 mL of n-butanol was added, and the reaction was carried out for 1 hour by microwave at 150° C. The reaction solution was evaporated to dryness under reduced pressure, and 0.80 g of a pale yellow gel-like crude product was obtained by column chromatography (dichloromethane / methanol system) in a yield of 47.3%. 1 H NMR (300 MHz, DMSO-d 6 ) δ 11.40 (s, 1H), 9.97 (s, 1H), 8.38 (s, 1H), 8.19 (s, 1H), 7.79 (s, 1H), 7.07 (s, 1H), 6.98 (s, 1H), 4.06 (q, J = 7.0 Hz, 2H), 3.17 (d, J = 56.4 Hz,3H), 2.88 - 2.64 (m, 2H), 2.38 (s, 1H), 2.07 - 1.59 (m, 3H), 1.22 - 0.93 (m, 3H). MS (ESI) m / z: 332 [M+H] + .

[0060] Step 8: Synthesis of (S)-1-(5-formylamino-1H-7-azaindole-4-amino)piperidine-3-carboxylic acid [ka]

[0061] At room temperature, (S)-1-(5-formylamino-1H-7-azaindole-4-amino)piperidine-3-carboxylate ethyl (0.70 g, 2.11 mmol) was dissolved in methanol, and 3N NaOH solution was added dropwise and reacted for 3 hours at 23° C. The organic solvent was removed by distillation, the aqueous phase was adjusted to about pH=4 with 1N dilute hydrochloric acid, evaporated to dryness under reduced pressure, methanol was added to the residual solid to dissolve it, inorganic salts were removed by filtration, and methanol was removed by distillation to obtain 0.52 g of a pale yellow solid in a yield of 81.3%.

[0062] Step 9: Synthesis of (S)-4-(3-(2,2,2-trifluoroethylcarbamoyl)piperidine-1-amino)-1H-7-azaindole-5-formamide [ka]

[0063] (S)-1-(5-formylamino-1H-7-azaindole-4-amino)piperidine-3-carboxylic acid (0.25 g, 0.82 mmol), trifluoroammonium hydrochloride (0.17 g, 1.23 mmol) and O-benzotriazole-N,N,N',N'-tetramethyluronium tetrafluoroborate (0.40 g, 1.23 mmol, TBTU) were added to a single-neck flask, 5 mL of DMF was added, triethylamine (0.48 g, 4.75 mmol) was added while stirring, and the mixture was stirred at room temperature overnight. 10 mL of water was added to the reaction solution, and the mixture was extracted with a mixed solvent of DCM:MeOH = 5:1 (10 mL x 5). The organic phases were combined, washed with saturated saline, evaporated to dryness under reduced pressure, and column chromatography (dichloromethane / methanol system) was used to obtain 0.022 g of a light yellow solid in 10% yield.1 H NMR (400 MHz, DMSO-d 6 ) δ 11.42 (br, 1H), 9.95 (br, 1H), 8.59 (br, 1H), 8.37 (s, 1H), 8.15(s, 1H), 7.78 (br, 1H), 7.04 (m, 2H), 3.87 (br, 2H), 3.11 (m, 2H), 2.71 (m, 1H), 2.37 (m, 2H), 1.82 (m, 2H), 1.67 (m, 1H), 1.34 (m, 1H). MS (ESI) m / z: 385 [M+H] + .

[0064] Example 2 Synthesis of (S)-4-(3-(2,2-difluoroethylcarbamoyl)piperidine-1-amino)-1H-7-azaindole-5-formamide [ka]

[0065] Step 1: Synthesis of (S)-t-butyl 3-((2,2-difluoroethyl)carbamoyl)piperidine-1-carboxylate [ka]

[0066] At room temperature, (S)-1-(t-butoxycarbonyl)piperidine-3-carboxylic acid (15.00g, 65.40mmol) was dissolved in DCM (400mL), triethylamine (16.50g, 163.50mmol) and TBTU (25.20g, 78.50mmol) were added and reacted for 1 hour, then transferred to a 0°C ice bath, 2,2-difluoro-O-amine hydrochloride (10.00g, 85.00mmol) was slowly added, stirred at 0°C for 1 hour, and reacted at room temperature for 5 hours to complete. The mixture was concentrated, diluted with water (200mL), and extracted three times with ethyl acetate (150mL). The organic phases were combined, washed three times with 100mL of saturated saline, concentrated, and subjected to column chromatography (PE / EA system) to obtain 16.06g of a colorless, transparent liquid in 84% yield. MS (ESI) m / z: 237.1 [M+H-56] + .

[0067] Step 2: Synthesis of (S)-N-(2,2-difluoroethyl)piperidine-3-formamide hydrochloride [ka]

[0068] (S)-3-((2,2-difluoroethyl)carbamoyl)piperidine-1-carboxylate t-butyl (16.06 g, 54.94 mmol) was dissolved in ethanol (150 mL), and 35% HCl / EtOH (30 mL) was slowly added dropwise at room temperature, and the reaction was completed for 3 hours. The solvent was removed under reduced pressure to obtain 11.43 g of a white solid in 91% yield. MS (ESI) m / z: 193.1 [M+H] + .

[0069] Step 3: Synthesis of (S)-N-(2,2-difluoroethyl)-1-nitrosopiperidine-3-formamide [ka]

[0070] At room temperature, (S)-N-(2,2-difluoroethyl)piperidine-3-formamide hydrochloride (11.43 g, 49.98 mmol) was dissolved in acetic acid (100 mL), and 50 mL of an aqueous solution in which sodium nitrite (4.49 g, 65.00 mmol) was dissolved was slowly added at 0°C. After the dropwise addition, the mixture was reacted at 0°C for 1 hour, and then the mixture was heated to room temperature and reacted overnight. After the reaction was completed, water (100 mL) was added to the reaction solution, and the mixture was extracted three times with ethyl acetate (80 mL). The organic phases were combined, washed with saturated aqueous sodium carbonate solution (80 mL), dried over anhydrous sodium sulfate, and concentrated. The mixture was mixed with silica gel and subjected to column chromatography (PE / EA) to obtain 9.80 g of a white solid in 89% yield. MS (ESI) m / z: 222.1 [M+H] + .

[0071] Step 4: Synthesis of (S)-1-amino-N-(2,2-difluoroethyl)piperidine-3-formamide hydrochloride [ka]

[0072] (S)-N-(2,2-difluoroethyl)-1-nitrosopiperidine-3-formamide (9.80 g, 44.30 mmol) was dissolved in methanol (40 mL), and zinc powder (8.70 g, 132.90 mmol) was slowly added at room temperature, and the mixture was cooled to -20 ° C under nitrogen gas protection and stirred for 10 minutes. Acetic acid (50 mL) was slowly added dropwise at -20 ° C, and the reaction was completed for 2 hours under nitrogen gas protection, and a solution of hydrochloric acid in ethyl acetate was added, stirred for 1 hour to form a salt, and evaporated to dryness under reduced pressure to store. The reaction solution was filtered, and the filtrate was directly mixed with silica gel and column chromatography (DCM / MeOH) was performed to obtain 8.60 g of a white solid, and a solution of hydrochloric acid in ethyl acetate was added, stirred for 1 hour to form a salt, and evaporated to dryness under reduced pressure to store. MS (ESI) m / z: 208.1 [M + H] + .

[0073] Step 5: Synthesis of (S)-4-(3-(2,2-difluoroethylcarbamoyl)piperidine-1-amino)-1H-7-azaindole-5-formamide [ka]

[0074] 4-Chloro-7-azaindole-5-formamide (0.10 g, 0.51 mmol) and (S)-1-amino-N-(2,2-difluoroethyl)piperidine-3-formamide hydrochloride (0.25 g, 1.02 mmol) were added to a microwave tube, 2 mL of n-butanol was added, and the mixture was reacted for 1 hour by microwave at 150° C. The reaction solution was filtered, the filtrate was spin-dried, and 0.014 g of a light yellow solid was obtained by column chromatography (dichloromethane / methanol system) in a yield of 7.5%. 1 H NMR (400 MHz, DMSO-d 6 ) δ 11.41 (br, 1H), 9.93 (br, 1H), 8.37 (s, 1H), 8.32 (br, 1H), 7.81 (br, 1H), 7.04 (m, 3H), 5.96 (t, J = 57.5 Hz, 1H), 3.44 (m, 2H), 3.12 (m, 2H), 2.68 (m, 1H), 2.33 (m, 2H), 1.78 (m, 2H), 1.65 (m, 1H), 1.32 (m, 1H). MS (ESI) m / z: 367 [M+H] + .

[0075] Example 3 Synthesis of (R)-4-(3-(2,2,2-trifluoroethylcarbamoyl)piperidine-1-amino)-1H-7-azaindole-5-formamide [ka]

[0076] The synthesis of the intermediate (R)-1-amino-N-(2,2,2-trifluoroethyl)piperidine-3-formamide hydrochloride was carried out by referring to the preparation method of the intermediate (S)-1-amino-N-(2,2-difluoroethyl)piperidine-3-formamide hydrochloride in Example 2, except that (S)-1-(t-butoxycarbonyl)piperidine-3-carboxylic acid in step (1) of Example 2 was replaced with (R)-1-(t-butoxycarbonyl)piperidine-3-carboxylic acid and 2,2-difluoro-O-amine hydrochloride was replaced with trifluoro-O-amine hydrochloride.

[0077] 4-Chloro-7-azaindole-5-formamide (0.10 g, 0.51 mmol), (R)-1-amino-N-(2,2,2-trifluoroethyl)piperidine-3-formamide hydrochloride (0.27 g, 1.02 mmol) were added to a microwave tube, 2 mL of n-butanol was added, and the mixture was reacted for 1 hour by microwave at 150° C. The reaction solution was filtered, the filtrate was spin-dried, and 0.015 g of a light yellow solid powder was obtained by column chromatography (dichloromethane / methanol system) in a yield of 7.6%. 1 H NMR (400 MHz, DMSO-d 6 ) δ 11.41 (br, 1H), 9.94 (br, 1H), 8.60 (br, 1H), 8.37 (s, 1H), 8.15 (br, 1H), 7.82 (br, 1H), 7.04 (m, 2H), 3.88 (br, 2H), 3.10 (m, 2H), 2.71 (m, 1H), 2.38 (m, 2H), 1.82 (m, 2H), 1.67 (m, 1H), 1.37 (m, 1H). MS (ESI) m / z: 385 [M+H] + .

[0078] Example 4 Synthesis of (R)-4-(3-(2,2-difluoroethylcarbamoyl)piperidine-1-amino)-1H-7-azaindole-5-formamide [ka]

[0079] (R)-1-amino-N-(2,2-difluoroethyl)piperidine-3-formamide hydrochloride was prepared by referring to the preparation method of (S)-1-amino-N-(2,2-difluoroethyl)piperidine-3-formamide in Example 2, except that (S)-1-(t-butoxycarbonyl)piperidine-3-carboxylic acid in step (1) of Example 2 was replaced with (R)-1-(t-butoxycarbonyl)piperidine-3-carboxylic acid.

[0080] 4-Chloro-7-azaindole-5-formamide (0.10 g, 0.51 mmol) and (R)-1-amino-N-(2,2-difluoroethyl)piperidine-3-formamide hydrochloride (0.25 g, 1.02 mmol) were added to a microwave tube, 2 mL of n-butanol was added, and the mixture was reacted for 1 hour in a microwave at 150° C. The reaction solution was filtered, the filtrate was spin-dried, and 0.035 g of a light yellow solid powder was obtained by column chromatography (dichloromethane / methanol system) in a yield of 18.7%. 1 H NMR (400 MHz, DMSO-d 6 ) δ 11.42 (br, 1H), 9.94 (br, 1H), 8.38 (s, 1H), 8.32 (br, 1H), 7.82 (br, 1H), 7.08 (m, 3H), 5.97 (t, J = 57.5 Hz, 1H), 3.46 (br, 2H), 3.12 (m, 2H), 2.68 (m, 1H), 2.37 (m, 2H), 1.80 (m, 2H), 1.67 (m, 1H), 1.34 (m, 1H). MS (ESI) m / z: 367 [M+H] + .

[0081] Example 5 Synthesis of (S)-4-(3-(n-propylcarbamoyl)piperidine-1-amino)-1H-7-azaindole-5-formamide [ka]

[0082] (S)-1-(5-formylamino-1H-7-azaindole-4-amino)piperidine-3-carboxylic acid (0.25g, 0.82mmol), n-propylamine (0.058g, 1.00mmol) and O-benzotriazole-N,N,N',N'-tetramethyluronium tetrafluoroborate (0.39g, 1.23mmol, TBTU) were added to a single-neck flask, 5mL of DMF was added, triethylamine (0.25g, 2.46mmol) was added with stirring, and the mixture was reacted at room temperature for 16 hours with stirring. The reaction solution was directly evaporated to dryness under reduced pressure, and 48mg of a light yellow solid was obtained by column chromatography (dichloromethane / methanol system) in a yield of 17%. 1 H NMR (300 MHz, DMSO-d 6 ) δ 11.44 (br, 1H), 9.97 (br, 1H), 8.38 (s, 1H), 7.85 (br, 2H), 7.05 (m, 3H), 3.11 (m, 2H), 2.98 (m, 2H), 2.60 (m, 1H), 2.30 (m, 2H), 1.72 (m, 3H), 1.38(m, 3H), 0.81 (t, J = 7.4 Hz, 3H). MS (ESI) m / z: 345 [M+H] + .

[0083] Example 6 Synthesis of (S)-4-(3-(cyclopropylcarbamoyl)piperidine-1-amino)-1H-7-azaindole-5-formamide [ka]

[0084] (S)-1-(5-formylamino-1H-7-azaindole-4-amino)piperidine-3-carboxylic acid (0.16 g, 0.53 mmol), cyclopropylamine (0.036 g, 0.63 mmol) and O-benzotriazole-N,N,N',N'-tetramethyluronium tetrafluoroborate (0.25 g, 0.80 mmol, TBTU) were added to a single-neck flask, 5 mL of DMF was added, triethylamine (0.16 g, 1.59 mmol) was added with stirring, and the mixture was reacted at room temperature for 16 hours with stirring. The reaction solution was directly evaporated to dryness under reduced pressure, and 45 mg of a light yellow solid was obtained by column chromatography (dichloromethane / methanol system) in a yield of 25%. 1 H NMR (300 MHz, DMSO-d 6 ) δ 11.52 (br, 1H), 10.03 (br, 1H), 8.39 (s, 1H), 7.93 (br, 2H), 7.09 (m, 2H), 7.01 (m, 1H), 3.09 (m, 2H), 2.59 (m, 2H), 2.33 (m, 2H), 1.80 (m, 2H), 1.65 (m, 1H), 1.36 (m,1H), 0.56 (dt, J = 4.6, 2.7 Hz, 2H), 0.36 (dt, J = 4.6, 2.7 Hz, 2H). MS (ESI) m / z: 343 [M+H] + .

[0085] Example 7 Synthesis of (S)-4-(3-(cyanoethylcarbamoyl)piperidine-1-amino)-1H-7-azaindole-5-formamide [ka]

[0086] (S)-1-(5-formylamino-1H-7-azaindole-4-amino)piperidine-3-carboxylic acid (0.25g, 0.82mmol), aminoacetonitrile hydrochloride (0.093g, 1.00mmol) and O-benzotriazole-N,N,N',N'-tetramethyluronium tetrafluoroborate (0.39g, 1.23mmol, TBTU) were added to a single-neck flask, 5mL of DMF was added, triethylamine (0.25g, 2.46mmol) was added with stirring, and the mixture was reacted at room temperature for 16 hours with stirring. The reaction solution was directly evaporated to dryness under reduced pressure, and 43mg of a light yellow solid was obtained by column chromatography (dichloromethane / methanol system) in a yield of 15%. 1 H NMR (300 MHz, DMSO-d 6 ) δ 11.50 (br, 1H), 10.01 (br, 1H), 8.67 (br, 1H), 8.38 (s, 1H), 7.84 (br, 1H), 7.05 (m, 3H), 4.11 (br, 2H), 3.18 (m, 2H), 2.66 (m, 1H), 2.38 (m, 2H), 1.77 (m, 3H), 1.30 (m, 1H). MS (ESI) m / z: 342 [M+H] + .

[0087] Example 8 (S)-1-(5-formylamino-1H-7-azaindole-4-amino)piperidine-3-carboxylate ethyl ester [ka]

[0088] (S)-1-(5-formylamino-1H-7-azaindole-4-amino)piperidine-3-carboxylate is an intermediate obtained in Step 7 of Example 1. 1 H NMR (300 MHz, DMSO-d 6) δ 11.40 (br, 1H), 9.97 (br, 1H), 8.38 (s, 1H), 8.19 (br, 1H), 7.79 (br, 1H), 7.07 (br, 1H), 6.98 (br, 1H), 4.06 (q, J = 7.0 Hz, 2H), 3.18 (m, 2H), 2.78 (m, 1H), 2.38 (m, 2H), 1.79 (m, 3H), 1.30 (m, 1H), 1.15(t, J = 4.0 Hz, 3H). MS (ESI) m / z: 332 [M+H] + .

[0089] Example 9 Synthesis of 4-(3-(2,2,2-trifluoroethylcarbamoyl)pyrrole-1-amino)-1H-7-azaindole-5-formamide [ka]

[0090] Step 1: Synthesis of 1-(t-butoxycarbonyl)pyrrole-3-carboxylic acid [ka]

[0091] At room temperature, 3-pyrrole carboxylic acid (1.00 g, 8.68 mmol) was dissolved in a mixed solvent consisting of 10 mL of THF and 5 mL of water, sodium bicarbonate (2.18 g, 0.026 mol) was added, and the temperature was lowered to 0°C. (Boc) 2 After adding O (2.08 g, 9.55 mmol) dropwise, the mixture was warmed to room temperature and reacted for 2 hours. 100 mL of EA was added to the reaction solution, extracted to remove impurities, the aqueous phase was adjusted to pH=4 with citric acid solution, DCM was added to extract, dried over anhydrous sodium sulfate, and concentrated to obtain 1.64 g of a white solid in 88% yield.

[0092] Step 2: Synthesis of 1-(t-butoxycarbonyl)pyrrole-3-(2,2,2-trifluoroethyl)formamide [ka]

[0093] At room temperature, 1-(t-butoxycarbonyl)pyrrole-3-carboxylic acid (1.64g, 7.62mmol) was added to 20mL of DMF, and O-benzotriazole-N,N,N',N'-tetramethyluronium tetrafluoroborate (3.67g, 11.43mmol, TBTU), trifluoroammonium hydrochloride (1.24g, 9.15mmol), and triethylamine (2.31g, 0.023mol) were added in sequence, and the mixture was reacted at room temperature for 16 hours with stirring. The reaction solution was poured into 50mL of water, extracted by adding 30mL of DCM, the aqueous phase was extracted twice with DCM, the organic phase was combined, washed with saturated saline, dried over anhydrous sodium sulfate, evaporated to dryness under reduced pressure, and purified by silica gel column chromatography (PE:EA=1:1) to obtain 1.90g of a pale yellow oil in 84% yield.

[0094] Step 3: Synthesis of N-(2,2,2-trifluoroethyl)pyrrole-3-formamide hydrochloride [ka]

[0095] 1-(t-butoxycarbonyl)pyrrole-3-(2,2,2-trifluoroethyl)formamide (1.90 g, 6.42 mmol) was dissolved in a solution of hydrochloric acid in ethyl acetate (20 mL) and reacted at room temperature for 16 hours with stirring. The solvent was directly removed from the reaction solution under reduced pressure to obtain 1.45 g of a viscous oily product with a crude product yield of 100%.

[0096] Step 4: 1-Nitroso-N-(2,2,2-trifluoroethyl)pyrrole-3-formyl Synthesis of amides [ka]

[0097] N-(2,2,2-trifluoroethyl)piperidine-3-formamide hydrochloride (1.45 g, 6.23 mmol) was added to a mixed solvent consisting of 20 mL of glacial acetic acid and 5 mL of water, cooled to 0°C, sodium nitrite (0.65 g, 9.35 mmol) was added dropwise, and the prepared 5 mL of aqueous solution was reacted for 1 hour while stirring at 0°C, and then heated to room temperature and reacted for 2 hours. Water was added to the reaction solution, and it was extracted three times with ethyl acetate. The organic phases were combined, washed with saturated saline, and evaporated to dryness under reduced pressure to obtain 1.25 g of a light yellow liquid with a crude product yield of 95%, which was directly put into the next reaction.

[0098] Step 5: Synthesis of 1-amino-N-(2,2,2-trifluoroethyl)pyrrole-3-formamide hydrochloride [ka]

[0099] 1-Nitroso-N-(2,2,2-trifluoroethyl)pyrrole-3-formamide (1.25g, 5.90mmol) was dissolved in 20mL of methanol, Zn (1.15g, 0.018mol) was added, the temperature was lowered to -5℃, glacial acetic acid (10mL) was slowly added dropwise, and the mixture was stirred at 0℃ for 0.5 hours, naturally warmed to room temperature, and reacted for 2 hours. The reaction solution was filtered, the filter cake was washed with methanol, the filtrate was evaporated to dryness under reduced pressure, 50mL of water was added to the residue, the pH of the solution was adjusted to 9 with sodium carbonate, and the mixture was extracted five times with a mixed solvent of DCM / MeOH (5:1), the organic phase was combined, washed once with saturated saline, then dried over anhydrous sodium sulfate, filtered, and concentrated to obtain a light yellow liquid, to which a solution of hydrochloric acid in ethyl acetate was added, and the mixture was stirred for 1 hour to make a salt, and the solvent was spin-dried to obtain 1.50g of a light yellow oil in 100% yield.

[0100] Step 6: Synthesis of 4-(3-(2,2,2-trifluoroethylcarbamoyl)pyrrole-1-amino)-1H-7-azaindole-5-formamide The preparation method of 4-(3-(2,2,2-trifluoroethylcarbamoyl)pyrrole-1-amino)-1H-7-azaindole-5-formamide was prepared by referring to the method in step (5) of Example 2, except that (S)-1-amino-N-(2,2-difluoroethyl)piperidine-3-formamide hydrochloride was replaced with 1-amino-N-(2,2,2-trifluoroethyl)pyrrole-3-formamide hydrochloride. 1 H NMR (300 MHz, DMSO-d 6 ) δ 11.40 (br, 1H), 9.97 (br ,1H), 8.62 (br, 1H), 8.36 (s, 1H), 7.77 (br, 1H), 7.04 (m, 3H), 3.95 (m, 2H), 3.23 - 3.02 (m, 3H), 2.69 (m, 2H), 2.07 (m, 2H). MS (ESI) m / z: 371 [M+H] + .

[0101] Example 10 Synthesis of 4-((3-oxa-8-azabicyclo[3.2.1]octan-8-yl)amino)-1H-7-azaindole-5-formamide [ka]

[0102] Step 1: Synthesis of 8-nitroso-3-oxa-8-azabicyclo[3.2.1]octane [ka]

[0103] At room temperature, 3-oxa-8-azabicyclo[3.2.1]octane hydrochloride (0.30 g, 2.00 mmol) was dissolved in 6 mL of acetic acid, and 3 mL of an aqueous solution in which sodium nitrite (0.21 g, 0.30 mmol) was dissolved was slowly added at 0°C. After the dropwise addition, the mixture was reacted at 0°C for 1 hour, and then heated to room temperature and reacted overnight. After the reaction was completed, 15 mL of water was added to the reaction solution, and the mixture was extracted three times with 15 mL of ethyl acetate. The organic phases were combined, washed with a saturated aqueous sodium carbonate solution, dried over anhydrous sodium sulfate, and concentrated. 0.24 g of a yellow solid was obtained by silica gel column chromatography (PE / EA) in a yield of 85%. MS (ESI) m / z: 143 [M+H] + .

[0104] Step 2: Synthesis of 3-oxa-8-azabicyclo[3.2.1]octan-8-amine [ka]

[0105] 8-Nitroso-3-oxa-8-azabicyclo[3.2.1]octane (0.24 g, 1.69 mmol) was dissolved in 4 mL of methanol, and zinc powder (0.33 g, 5.06 mmol) was slowly added at room temperature. The mixture was cooled to -20°C under nitrogen gas protection and stirred for 10 minutes. 4 mL of acetic acid was slowly added dropwise at -20°C and reacted with stirring for 2 hours. The reaction solution was filtered, and the filtrate was directly evaporated to dryness, and 0.20 g of a white solid oil mixture was obtained in a 93% yield by silica gel column chromatography (DCM / MeOH). MS (ESI) m / z: 129 [M+H] + .

[0106] Step 3: Synthesis of 4-((3-oxa-8-azabicyclo[3.2.1]octan-8-yl)amino)-1H-7-azaindole-5-formamide The preparation method of 4-((3-oxa-8-azabicyclo[3.2.1]octan-8-yl)amino)-1H-7-azaindole-5-formamide was prepared by referring to the method in step (5) of Example 2, except that (S)-1-amino-N-(2,2-difluoroethyl)piperidine-3-formamide hydrochloride was replaced with 3-oxa-8-azabicyclo[3.2.1]octan-8-amine.

[0107] 1 H NMR (300 MHz, DMSO-d 6 ) δ 11.41 (br, 1H), 10.44 (br, 1H), 8.38 (s, 1H), 8.33 (br, 1H), 7.23 (br, 1H), 7.06 (m, 2H), 3.77 (d, J = 10.7 Hz, 2H), 3.57 (d, J = 10.6 Hz, 2H), 3.27 (m, 2H), 1.94 (q, J = 10.5, 9.2 Hz, 4H). MS (ESI) m / z:288 [M+H] + .

[0108] Example 11 Synthesis of 4-((3-hydroxy-8-azabicyclo[3.2.1]octan-8-yl)amino)-1H-7-azaindole-5-formamide [ka]

[0109] Step 1: Synthesis of (1R,3r,5S)-8-nitroso-8-azabicyclo[3.2.1]octan-3-ol [ka]

[0110] At room temperature, (1R,3r,5S)-8-azabicyclo[3.2.1]octan-3-ol hydrochloride (0.50 g, 3.94 mmol) was dissolved in acetic acid (10 mL), and 5 mL of an aqueous solution in which sodium nitrite (0.54 g, 7.88 mmol) was dissolved was slowly added at 0°C. After the dropwise addition, the mixture was reacted at 0°C for 1 hour, and then the mixture was heated to room temperature and reacted overnight. After the reaction was completed, water (15 mL) was added to the reaction solution, and the mixture was extracted three times with ethyl acetate (15 mL). The organic phases were combined, washed with saturated aqueous sodium carbonate solution (20 mL), dried over anhydrous sodium sulfate, concentrated, and subjected to silica gel column chromatography (PE / EA) to obtain 0.59 g of a yellow solid in a yield of 96%. MS (ESI) m / z: 157.1 [M+H] + .

[0111] Step 2: Synthesis of (1R,3r,5S)-8-amino-8-azabicyclo[3.2.1]octan-3-ol [ka]

[0112] (1R,3r,5S)-8-nitroso-8-azabicyclo[3.2.1]octan-3-ol (0.51g, 3.27mmol) was dissolved in methanol (6mL), zinc powder (0.64g, 9.81mmol) was slowly added at room temperature, and the mixture was cooled to -20°C under nitrogen gas protection and stirred for 10 minutes. Acetic acid (8mL) was slowly added dropwise at -20°C, and the reaction was completed at 0°C for 2 hours under nitrogen gas protection. The reaction solution was filtered, the pH of the filtrate was adjusted to 8 with sodium carbonate, concentrated, and subjected to silica gel column chromatography (DCM / MeOH) to obtain 0.57g of a white solid oil mixture in a yield of more than 100%. MS (ESI) m / z: 143.1[M+H] + .

[0113] Step 3: Synthesis of ((3-hydroxy-8-azabicyclo[3.2.1]octan-8-yl)amino)-1H-7-azaindole-5-formamide The preparation method of ((3-hydroxy-8-azabicyclo[3.2.1]octan-8-yl)amino)-1H-7-azaindole-5-formamide was prepared by referring to the method in step (5) of Example 2, except that (S)-1-amino-N-(2,2-difluoroethyl)piperidine-3-formamide hydrochloride was replaced with (1R,3r,5S)-8-amino-8-azabicyclo[3.2.1]octan-3-ol.

[0114] 1 H NMR (400 MHz, methanol-d 4 ) δ 8.31 (s, 1H), 7.71 (br, 1H), 7.34 (d, J = 3.5 Hz, 1H), 7.08 (br, 2H), 3.40 (t, J = 3.6 Hz, 1H), 2.37 - 2.31 (m, 2H), 2.25 - 2.10 (m, 3H), 2.05 (dd, J = 12.3, 6.4 Hz, 1H), 1.94 (d, J = 3.5 Hz, 1H), 1.91 (m, 1H), 1.61 (t, J = 13.1 Hz, 2H). MS (ESI) m / z:302 [M+H] + .

[0115] Example 12 4-(N-Boc-5-amino-hexahydropyrrolo[3,4-c]pyrrole-2(1H Synthesis of ))-1H-7-Azaindole-5-formamide [ka]

[0116] Step 1: Synthesis of N-Boc-5-nitroso-hexahydropyrrolo[3,4-c]pyrrole-2(1H) [ka]

[0117] N-Boc-hexahydropyrrolo[3,4-c]pyrrole-2(1H) (2.12 g, 10 mmol) was added to a mixed solvent consisting of 20 mL of glacial acetic acid and 5 mL of water, cooled to 0°C, and 5 mL of an aqueous solution containing sodium nitrite (1.03 g, 15 mmol) was added dropwise, stirred at 0°C for 1 hour, then allowed to naturally warm to room temperature and react for 2 hours. Water was added to the reaction solution, which was then extracted three times with ethyl acetate. The organic phases were combined and washed with saturated saline, and the resulting white solid (2.41 g) was evaporated to dryness under reduced pressure and directly introduced into the next step.

[0118] Step 2: Synthesis of N-Boc-5-amino-hexahydropyrrolo[3,4-c]pyrrole-2(1H) [ka]

[0119] N-Boc-5-nitroso-hexahydropyrrolo[3,4-c]pyrrole-2(1H) (2.41g, 10mmol) was dissolved in 20mL of methanol, Zn (1.95g, 0.03mol) was added, the temperature was lowered to -5℃, 10mL of glacial acetic acid was slowly added dropwise, and the mixture was stirred at 0℃ for 0.5 hours, naturally warmed to room temperature, and reacted for 2 hours. The reaction solution was filtered, the filter cake was washed with methanol, the filtrate was evaporated to dryness under reduced pressure, 50mL of water was added to dissolve, the pH of the solution was adjusted to 9 with sodium carbonate, and the mixture was extracted five times with a mixed solvent of DCM / MeOH (5:1), the organic phases were combined, washed with saturated saline, dried over anhydrous sodium sulfate, and the solvent was distilled off to obtain 1.76g of a white solid in 81.8% yield.

[0120] Step 3: Synthesis of 4-(N-Boc-5-amino-hexahydropyrrolo[3,4-c]pyrrole-2(1H))-1H-7-azaindole-5-formamide The preparation method of 4-(N-Boc-5-amino-hexahydropyrrolo[3,4-c]pyrrole-2(1H))-1H-7-azaindole-5-formamide was prepared by referring to the method in step (5) of Example 2, except that (S)-1-amino-N-(2,2-difluoroethyl)piperidine-3-formamide hydrochloride was replaced with N-Boc-5-amino-hexahydropyrrolo[3,4-c]pyrrole-2(1H).

[0121] 1 H NMR (400 MHz, DMSO-d 6 ) δ 11.38 (br, 1H), 9.86 (br, 1H), 8.36 (s, 1H), 7.78 (br, 1H), 7.15 - 6.58 (m, 3H), 3.57 (m, 2H), 3.09 (m, 2H), 2.88 - 2.51 (m, 4H), 1.45 (s, 9H), 1.18 (m, 1H), 0.77 (m, 1H). MS (ESI) m / z: 387 [M+H] + .

[0122] Example 13 Synthesis of 4-(5-acetylhexahydropyrrolo[3,4-c]pyrrole-2(1H))-1H-7-azaindole-5-formamide [ka]

[0123] Step 1: Synthesis of 1-(5-nitrosohexahydropyrrolo[3,4-c]pyrrol-2(1H))ethan-1-one [ka]

[0124] 1-(hexahydropyrrolo[3,4-c]pyrrole-2(1H))acetaldehyde hydrochloride (1.50 g, 7.87 mmol) was added to a mixed solvent consisting of 20 mL of glacial acetic acid and 5 mL of water, cooled to 0°C, and 5 mL of an aqueous solution containing sodium nitrite (0.81 g, 11.80 mmol) was added dropwise, stirred at 0°C for 1 hour, naturally warmed to room temperature, and reacted for 2 hours. Water was added to the reaction solution, which was then extracted three times with ethyl acetate. The organic phases were combined, washed with saturated saline, and the solvent was distilled off under reduced pressure to obtain 1.44 g of a pale yellow liquid in 100% yield, which was directly used in the next step.

[0125] Step 2: Synthesis of 1-(5-aminohexahydropyrrolo[3,4-c]pyrrole-2(1H))acetaldehyde hydrochloride [ka]

[0126] 1-(5-Nitrosohexahydropyrrolo[3,4-c]pyrrole-2(1H))acetaldehyde (1.44 g, 7.87 mmol) was dissolved in 20 mL of methanol, Zn powder (1.53 g, 0.023 mol) was added, the temperature was lowered to -5°C, glacial acetic acid (10 mL) was slowly added dropwise, and the mixture was stirred at 0°C for 0.5 hours, naturally warmed to room temperature, and reacted for 2 hours. The reaction solution was filtered, the filter cake was washed with methanol, the filtrate was evaporated to dryness under reduced pressure, 50mL of water was added to the residue to dissolve it, the pH of the solution was adjusted to 9 with sodium carbonate, and the mixture was extracted five times with a mixed solvent of DCM / MeOH (5:1). The organic phases were combined, washed with saturated saline, dried over anhydrous sodium sulfate, and the solvent was evaporated under reduced pressure. A solution of hydrochloric acid in ethyl acetate was added to the resulting pale yellow liquid, which was stirred for 0.5 hours to form a salt, and the solvent was evaporated to obtain 2.00g of a pale yellow oil with a crude product yield of 100%.

[0127] Step 3: Synthesis of 4-(5-acetylhexahydropyrrolo[3,4-c]pyrrole-2(1H))-1H-7-azaindole-5-formamide The preparation method of 4-(5-acetylhexahydropyrrolo[3,4-c]pyrrole-2(1H))-1H-7-azaindole-5-formamide was prepared by referring to the method in step (5) of Example 2, except that (S)-1-amino-N-(2,2-difluoroethyl)piperidine-3-formamide hydrochloride was replaced with 1-(5-aminohexahydropyrrolo[3,4-c]pyrrole-2(1H))acetaldehyde hydrochloride.

[0128] 1 H NMR (400 MHz, DMSO-d 6 ) δ 11.40 (br, 1H), 9.85 (br, 1H), 8.36 (s, 1H), 7.88 (br, 1H), 7.12 (br, 1H), 7.03 (br, 1H), 6.85 (br, 1H), 3.67 (m, 2H), 3.14 - 2.62 (m, 6H), 2.00 (s, 3H), 1.28 (m, 1H), 0.86 (m, 1H). MS (ESI) m / z: 329 [M+H] + .

[0129] Example 14 Synthesis of 4-(1-oxo-2,8-diazaspiro[4.5]decane-8-amino)-1H-7-azaindole-5-formamide [ka]

[0130] Step 1: Synthesis of 8-nitroso-2,8-diazaspiro[4.5]decan-1-one [ka]

[0131] 2,8-Diazaspiro[4.5]decan-1-one (2.08g, 20mmol) was added to a mixed solvent consisting of 20mL of glacial acetic acid and 5mL of water, cooled to 0℃, and 10mL of an aqueous solution containing sodium nitrite (2.07g, 30mmol) was added dropwise, stirred at 0℃ for 1 hour, naturally warmed to room temperature, and reacted for 2 hours. Water was added to the reaction solution, extracted three times with ethyl acetate, the organic phases were combined, washed with saturated saline, and evaporated to dryness under reduced pressure, and the resulting crude product (2.47g) was directly added to the next step and reacted.

[0132] Step 2: Synthesis of 8-amino-2,8-diazaspiro[4.5]decan-1-one hydrochloride [ka]

[0133] 8-Nitroso-2,8-diazaspiro[4.5]decan-1-one (2.47g, 20mmol) was dissolved in 20mL of methanol, Zn (3.90g, 0.06mol) was added, the temperature was lowered to -5℃, 10mL of glacial acetic acid was slowly added dropwise, and the mixture was stirred at 0℃ for 0.5 hours, naturally warmed to room temperature, and reacted for 2 hours. The reaction solution was filtered, the filter cake was washed with methanol, the filtrate was evaporated to dryness under reduced pressure, an ethanol solution of hydrochloric acid was added and stirred for 0.5 hours, the solvent was distilled off under reduced pressure, and the mixture was pulped with ethyl acetate, filtered, and the filter cake was dried to obtain 2.12g of a light yellow solid in a yield of 93%.

[0134] Step 3: Synthesis of 4-(1-oxo-2,8-diazaspiro[4.5]decane-8-amino)-1H-7-azaindole-5-formamide The preparation method of 4-(1-oxo-2,8-diazaspiro[4.5]decane-8-amino)-1H-7-azaindole-5-formamide was prepared by referring to the method in step (5) of Example 2, except that (S)-1-amino-N-(2,2-difluoroethyl)piperidine-3-formamide hydrochloride was replaced with 8-amino-2,8-diazaspiro[4.5]decane-1-one hydrochloride.

[0135] 1 H NMR (400 MHz, DMSO-d 6 ) δ 11.39 (br, 1H), 9.92 (br, 1H), 8.37 (s, 1H), 7.64 (br, 2H), 7.05 (m, 3H), 3.53 (m, 2H), 3.19 (m, 2H), 3.05 (m, 2H), 1.97 (m, 2H), 1.91 (m, 2H), 1.49 (m, 2H). MS (ESI) m / z: 329 [M+H] + .

[0136] Example 15 Synthesis of 4-(8-(cyclopropylcarbonyl)-3,8-diazabicyclo[3.2.1]octane-3-amino)-1H-7-azaindole-5-formamide [ka]

[0137] Step 1: 8-Cyclopropylcarbonyl (3-nitroso-3,8-diazabicyclo[3. 2.1]Octane synthesis [ka]

[0138] At room temperature, 8-cyclopropylcarbonyl-3,8-diazabicyclo[3.2.1]octane hydrochloride (1.11 g, 5.00 mmol) was added to a mixed solution consisting of 20 mL of glacial acetic acid and 5 mL of acetic acid, the temperature was lowered to 0°C, 5 mL of an aqueous solution containing dissolved sodium nitrite (0.53 g, 7.68 mmol) was added dropwise, the mixture was stirred at 0°C for 1 hour, and the mixture was allowed to naturally warm to room temperature and react for 2 hours. Water was added to the reaction solution, which was then extracted three times with ethyl acetate. The organic phases were combined, washed with saturated saline, and the solvent was distilled off under reduced pressure. The resulting pale yellow liquid (1.04 g) was used directly in the reaction of the next step.

[0139] Step 2: Synthesis of 8-cyclopropylcarbonyl (3-amino-3,8-diazabicyclo[3.2.1]octane [ka]

[0140] At room temperature, 8-cyclopropylcarbonyl(3-nitroso-3,8-diazabicyclo[3.2.1]octane (1.04g, 5.00mmol) was dissolved in 20mL of methanol, Zn powder (1.00g, 15.00mmol) was added, the temperature was lowered to -5℃, 10mL of glacial acetic acid was slowly added dropwise, and then the mixture was stirred at 0℃ for 0.5 hours, naturally warmed to room temperature and reacted for 2 hours, the reaction liquid was filtered, the filter cake was rinsed with methanol, the filtrate was collected, and after evaporation to dryness under reduced pressure, 50mL of water was added to dissolve, the pH was adjusted to 9 with aqueous sodium carbonate solution, and the mixture was extracted 5 times with a mixed solvent of DCM / MeOH (5:1), the organic phases were combined, washed with saturated saline, dried over anhydrous sodium sulfate, and the solvent was distilled off under reduced pressure to obtain 0.98g of a light yellow oil.

[0141] Step 3: Synthesis of 4-(8-(cyclopropylcarbonyl)-3,8-diazabicyclo[3.2.1]octane-3-amino)-1H-7-azaindole-5-formamide The preparation method of 4-(8-(cyclopropylcarbonyl)-3,8-diazabicyclo[3.2.1]octane-3-amino)-1H-7-azaindole-5-formamide was prepared by referring to the method in step (5) of Example 2, except that (S)-1-amino-N-(2,2-difluoroethyl)piperidine-3-formamide hydrochloride was replaced with 8-cyclopropylcarbonyl(3-amino-3,8-diazabicyclo[3.2.1]octane.

[0142] 1 H NMR (400 MHz, DMSO-d 6) δ 11.44 (br, 1H), 10.47 (br, 1H), 8.40 (s, 1H), 7.83 (br, 1H), 7.23 (br, 1H), 7.10 (m, 2H), 4.20 (d, J = 12.6 Hz, 1H), 4.10 (d, J = 13.9 Hz, 1H), 3.54 (d, J = 13.3 Hz, 1H), 3.43 (m, 2H), 2.98 (d, J = 13.7 Hz, 1H), 1.96 (m, 3H), 1.72 (m, 1H), 1.58 (m, 1H), 0.76 (m, 4H). MS (ESI) m / z: 355 [M+H] + .

[0143] Example 16 Synthesis of 4-(2-methyl-5-(2,2,2-trifluoroethylcarbamoyl)piperidine-1-amino)-1H-7-azaindole-5-formamide [ka]

[0144] Step 1: Synthesis of 6-methyl-1-nitroso-N-(2,2,2-trifluoroethyl)piperidine-3-formamide [ka]

[0145] 6-Methyl-N-(2,2,2-trifluoroethyl)piperidine-3-formamide (2.24 g, 10.00 mmol) was added to a mixed solvent consisting of 20 mL of glacial acetic acid and 10 mL of water, cooled to 0°C, and then 10 mL of an aqueous solution containing sodium nitrite (1.03 g, 15.00 mmol) was added dropwise, stirred at 0°C for 1 hour, naturally warmed to room temperature, and reacted for 1 hour. Water was added to the reaction solution, extracted three times with DCM, the organic phases were combined, washed three times with saturated sodium chloride, and evaporated to dryness under reduced pressure, and the resulting 2.53 g of pale yellow solid was directly added to the next step and reacted.

[0146] Step 2: Synthesis of 6-methyl-1-amino-N-(2,2,2-trifluoroethyl)piperidine-3-formamide [ka]

[0147] At room temperature, 6-methyl-1-nitroso-N-(2,2,2-trifluoroethyl)piperidine-3-formamide (2.53g, 10.00mmol) was dissolved in 20mL of methanol, Zn powder (2.40g, 40.00mmol) was added, the temperature was lowered to -5℃, 10mL of glacial acetic acid was slowly added dropwise, then the mixture was stirred at 0℃ for 0.5 hours, and the mixture was allowed to naturally warm to room temperature and react for 2 hours. The reaction solution was filtered, the filter cake was washed with methanol, and the filtrate was evaporated to dryness under reduced pressure. After adding 50mL of water to dissolve, the pH of the solution was adjusted to 9 with sodium carbonate, and the mixture was extracted 5 times with a mixed solvent of DCM / MeOH (5:1). The organic phases were combined, washed with saturated saline, dried over anhydrous sodium sulfate, and evaporated to dryness under reduced pressure to obtain 2.11g of a light yellow oil with a yield of 88%.

[0148] Step 3: Synthesis of 4-(2-methyl-5-(2,2,2-trifluoroethylcarbamoyl)piperidine-1-amino)-1H-7-azaindole-5-formamide The preparation method of 4-(2-methyl-5-(2,2,2-trifluoroethylcarbamoyl)piperidine-1-amino)-1H-7-azaindole-5-formamide was prepared by referring to the method in step (5) of Example 2, except that (S)-1-amino-N-(2,2-difluoroethyl)piperidine-3-formamide hydrochloride was replaced with 6-methyl-1-amino-N-(2,2,2-trifluoroethyl)piperidine-3-formamide.

[0149] 1 H NMR (400 MHz, methanol-d 4) δ 8.34 (s, 1H), 7.24 (d, J = 3.4 Hz, 1H), 7.06 (d, J = 3.4 Hz, 1H), 3.89 (m, 2H), 3.28 (m, 1H), 2.83 (m, 1H), 2.66 (m, 1H), 2.52 (m, 1H), 2.29 - 2.00 (m, 1H), 1.94 (m, 1H), 1.60 (m, 2H), 1.11 (d, J = 6.1 Hz, 3H). MS (ESI) m / z: 399 [M+H] + .

[0150] Example 17 Synthesis of 4-(8-(2,2,2-trifluoroethylcarbamoyl)-3,8-diazabicyclo[3.2.1]octane-3-amino)-1H-7-azaindole-5-formamide [ka]

[0151] Step 1: Synthesis of 3-nitroso-N-(2,2,2-trifluoroethyl)-3,8-diazabicyclo[3.2.1]octane-8-formamide [ka]

[0152] N-(2,2,2-trifluoroethyl)-3,8-diazabicyclo[3.2.1]octane-8-formamide hydrochloride (3.51 g, 10.00 mmol) was added to a mixed solvent consisting of 40 mL of glacial acetic acid and 10 mL of water, cooled to 0°C, and 10 mL of an aqueous solution containing sodium nitrite (1.04 g, 15.00 mmol) was added dropwise, stirred at 0°C for 1 hour, allowed to warm to room temperature, and reacted for 1 hour. Water was added to the reaction solution, extracted three times with DCM, the organic phases were combined, washed with saturated saline, and evaporated to dryness under reduced pressure, and the resulting pale yellow solid (2.66 g) was used directly in the next reaction step.

[0153] Step 2: Synthesis of 3-amino-N-(2,2,2-trifluoroethyl)-3,8-diazabicyclo[3.2.1]octane-8-formamide [ka]

[0154] At room temperature, 3-nitroso-N-(2,2,2-trifluoroethyl)-3,8-diazabicyclo[3.2.1]octane-8-formamide (2.66 g, 10.00 mmol) was dissolved in 20 mL of methanol, Zn (2.64 g, 40.00 mmol) was added, the temperature was lowered to -5 ° C, glacial acetic acid (10 mL) was slowly added dropwise, and the mixture was stirred at 0 ° C for 0.5 hours, naturally warmed to room temperature, and reacted for 16 hours. The reaction solution was filtered, the filter cake was washed with methanol, the filtrate was evaporated to dryness under reduced pressure, 50 mL of water was added to dissolve, the pH of the solution was adjusted to 9 with sodium carbonate, and the mixture was extracted five times with a DCM / MeOH (5:1) mixed solvent, the organic phases were combined, washed with saturated saline, dried with anhydrous sodium sulfate, and evaporated to dryness under reduced pressure to obtain 2.41 g of a light yellow oil in a yield of 95%.

[0155] Step 3: Synthesis of 4-(8-(2,2,2-trifluoroethylcarbamoyl)-3,8-diazabicyclo[3.2.1]octane-3-amino)-1H-7-azaindole-5-formamide The preparation method of 4-(8-(2,2,2-trifluoroethylcarbamoyl)-3,8-diazabicyclo[3.2.1]octane-3-amino)-1H-7-azaindole-5-formamide was prepared by referring to the method in step (5) of Example 2, except that (S)-1-amino-N-(2,2-difluoroethyl)piperidine-3-formamide hydrochloride was replaced with 3-amino-N-(2,2,2-trifluoroethyl)-3,8-diazabicyclo[3.2.1]octane-8-formamide.

[0156] 1 H NMR (400 MHz, methanol-d 4) δ 8.35 (s, 1H), 7.38 (d, J = 3.5 Hz, 1H), 7.10 (d, J = 3.5 Hz, 1H), 3.92 (m, 2H), 3.89 - 3.81 (m, 2H), 3.51 (m, 2H), 3.40 (m, 2H), 2.21 (m, 2H), 1.83 (m, 2H). MS (ESI) m / z: 412 [M+H] + .

[0157] Example 18 Synthesis of 4-(8-(2,2,2-trifluoroacetyl)-3,8-diazabicyclo[3.2.1]octane-3-amino)-1H-7-azaindole-5-formamide [ka]

[0158] Step 1: Synthesis of t-butyl 8-nitroso-3,8-diazabicyclo[3.2.1]octane-3-carboxylate [ka]

[0159] 8-Boc-3,8-diazabicyclo[3.2.1]octane (1.06g, 5.00mmol) was added to a mixed solvent consisting of 20mL of glacial acetic acid and 5mL of water, cooled to 0℃, and then 10mL of an aqueous solution containing sodium nitrite (0.52g, 15.00mmol) was added dropwise, stirred at 0℃ for 1 hour, allowed to naturally warm to room temperature, and reacted for 2 hours. Water was added to the reaction solution, which was then extracted three times with ethyl acetate. The organic phases were combined, washed with saturated saline, and evaporated to dryness under reduced pressure. The resulting crude product (1.21g) was directly added to the next step.

[0160] Step 2: Synthesis of t-butyl 8-amino-3,8-diazabicyclo[3.2.1]octane-3-carboxylate [ka]

[0161] 8-Nitroso-3,8-diazabicyclo[3.2.1]octane-3-carboxylate t-butyl (1.21g, 5.00mmol) was dissolved in 20mL of methanol contained in a four-neck flask, Zn powder (1.30g, 20.00mmol) was added, the temperature was lowered to -5℃, 10mL of glacial acetic acid was slowly added dropwise, and the mixture was stirred at 0℃ for 0.5 hours, naturally warmed to room temperature, and reacted for 2 hours. The reaction liquid was filtered, the filter cake was washed with methanol, and the solvent was distilled off under reduced pressure to obtain 1.14g of a pale yellow viscous liquid in a yield of 99%.

[0162] Step 3: Synthesis of 4-(8-(2,2,2-trifluoroacetyl)-3,8-diazabicyclo[3.2.1]octane-3-amino)-1H-7-azaindole-5-formamide Referring to the method in step 7 of Example 1, (S)-1-aminopiperidine-3-carboxylate ethyl hydrochloride in step 7 of Example 1 was replaced with t-butyl 8-amino-3,8-diazabicyclo[3.2.1]octane-3-carboxylate, and acylated with boc deprotection to obtain 4-(8-(2,2,2-trifluoroacetyl)-3,8-diazabicyclo[3.2.1]octane-3-amino)-1H-7-azaindole-5-formamide.

[0163] 1 H NMR (400 MHz, methanol-d 4 ) δ 8.53 (s, 1H), 7.38 (d, J = 3.5 Hz, 1H), 7.32 (d, J = 1.8 Hz, 1H), 4.25 (m, 2H), 3.42 (m, 2H), 3.14 (m, 2H), 2.47 (m, 2H), 2.30 (m, 2H). MS (ESI) m / z: 383 [M+H] + .

[0164] Example 19 Synthesis of 4-(4-(2,2,2-trifluoroethylcarbamoyl)piperidine-1-amino)-1H-7-azaindole-5-formamide [ka]

[0165] Step 1: Synthesis of 1-nitroso-N-(2,2,2-trifluoroethyl)piperidine-4-formamide [ka]

[0166] N-(2,2,2-trifluoroethyl)piperidine-4-formamide hydrochloride (2.10 g, 10.00 mmol) was added to a mixed solvent consisting of 20 mL of glacial acetic acid and 5 mL of water, and the mixture was cooled to 0°C, and then 5 mL of an aqueous solution containing sodium nitrite (1.04 g, 15.00 mmol) was added dropwise, stirred at 0°C for 1 hour, allowed to naturally warm to room temperature, and reacted for 2 hours. Water was added to the reaction solution, which was then extracted three times with ethyl acetate. The organic phases were combined, washed with saturated saline, and evaporated to dryness under reduced pressure. The resulting pale yellow solid (2.39 g) was directly added to the next step and reacted.

[0167] Step 2: Synthesis of 1-amino-N-(2,2,2-trifluoroethyl)piperidine-4-formamide hydrochloride [ka]

[0168] 1-Nitroso-N-(2,2,2-trifluoroethyl)piperidine-4-formamide (2.39g, 10.00mmol) was dissolved in 20mL of methanol, Zn powder (2.64g, 40.00mmol) was added, the temperature was lowered to -5℃, 10mL of glacial acetic acid was slowly added dropwise, and the mixture was stirred at 0℃ for 0.5 hours, then allowed to naturally warm to room temperature and react for 2 hours. The reaction solution was filtered, the filter cake was washed with methanol, the filtrate was evaporated to dryness under reduced pressure, 10mL of hydrochloric acid in ethanol was added to the residue, the mixture was stirred for 10 minutes, and then evaporated to dryness under reduced pressure to obtain 2.61g of a pale yellow viscous liquid in a 100% yield.

[0169] Step 3: Synthesis of 4-(4-(2,2,2-trifluoroethylcarbamoyl)piperidine-1-amino)-1H-7-azaindole-5-formamide The preparation method of 4-(4-(2,2,2-trifluoroethylcarbamoyl)piperidine-1-amino)-1H-7-azaindole-5-formamide was prepared by referring to the method in step (5) of Example 2, except that (S)-1-amino-N-(2,2-difluoroethyl)piperidine-3-formamide hydrochloride was replaced with 1-amino-N-(2,2,2-trifluoroethyl)piperidine-4-formamide hydrochloride.

[0170] 1 H NMR (400 MHz, DMSO-d 6 ) δ 11.41 (br, 1H), 9.95 (br, 1H), 8.59 (br, 1H), 8.37 (s, 1H), 7.83 (br, 1H), 7.08-7.03 (m, 3H),3.88 (br, 2H), 3.10 (m, 2H), 2.71 (m, 1H), 2.42 (m, 2H), 1.80 (m, 2H), 1.69 (m, 1H), 1.36 (m, 1H). MS (ESI) m / z: 385 [M+H] + .

[0171] Example 20 Synthesis of (S)-4-(3-(3,3-difluorocyclobutylcarbamoyl)piperidine-1-amino)-1H-7-azaindole-5-formamide [ka]

[0172] Step 1: Synthesis of (S)-N-(3,3-difluorocyclobutyl)-1-nitrosopiperidine-3-formamide [ka]

[0173] (S)-N-(3,3-difluorocyclobutyl)piperidine-3-formamide hydrochloride (2.18 g, 10.00 mmol) was added to a mixed solvent consisting of 20 mL of glacial acetic acid and 5 mL of water, and the mixture was cooled to 0°C, and then 5 mL of an aqueous solution containing sodium nitrite (1.04 g, 15.00 mmol) was added dropwise, stirred at 0°C for 1 hour, and then allowed to naturally warm to room temperature and react for 2 hours. Water was added to the reaction solution, which was then extracted three times with ethyl acetate. The organic phases were combined, washed with saturated saline, and evaporated to dryness under reduced pressure to obtain 2.47 g of a pale yellow solid in 100% yield, which was directly added to the next reaction.

[0174] Step 2: Synthesis of (S)-N-(3,3-difluorocyclobutyl)-1-nitrosopiperidine-3-formamide hydrochloride [ka]

[0175] 1-Nitroso-N-(2,2,2-trifluoroethyl)piperidine-4-formamide (2.47g, 10.00mmol) was dissolved in 20mL of methanol, Zn powder (2.64g, 40.00mmol) was added, the temperature was lowered to -5℃, 10mL of glacial acetic acid was slowly added dropwise, and the mixture was stirred at 0℃ for 0.5 hours, naturally warmed to room temperature, and reacted for 2 hours. The reaction liquid was filtered, the filter cake was washed with methanol, the filtrate was evaporated to dryness under reduced pressure, 10mL of hydrochloric acid in ethanol was added to the residue, stirred for 10 minutes, and evaporated to dryness under reduced pressure to obtain 2.69g of a pale yellow viscous liquid in 100% yield.

[0176] Step 3: Synthesis of (S)-4-(3-(3,3-difluorocyclobutylcarbamoyl)piperidine-1-amino)-1H-7-azaindole-5-formamide The preparation method of (S)-4-(3-(3,3-difluorocyclobutylcarbamoyl)piperidine-1-amino)-1H-7-azaindole-5-formamide was prepared by referring to the method in step (5) of Example 2, except that (S)-1-amino-N-(2,2-difluoroethyl)piperidine-3-formamide hydrochloride was replaced with (S)-N-(3,3-difluorocyclobutyl)-1-nitrosopiperidine-3-formamide hydrochloride.

[0177] 1 H NMR (400 MHz, methanol-d 4 ) δ 8.32 (s, 1H), 7.18 (br, 1H), 7.07 (d, J = 2.4 Hz, 1H), 4.12 (m, 1H), 2.90 (m, 2H), 2.74 (m, 1H), 2.54 (m, 4H), 2.22 (t, J = 7.3 Hz, 1H), 2.06 (m, 1H), 1.86 (m, 3H), 1.62 (m, 1H). MS (ESI) m / z: 393 [M+H] + .

[0178] Example 21 Synthesis of ethyl (S)-1-(5-cyano-1H-7-azaindole-4-amino)piperidine-3-carboxylate [ka]

[0179] Step 1: Synthesis of 4-chloro-1-(triisopropylsilyl)-1H-pyrrolo[2,3-b]pyridine-5-carbonitrile [ka]

[0180] Under the protection of nitrogen gas, 4-chloro-1H-pyrrolo[2,3-b]pyridine-5-carbonyl Tolyl (1.00 g, 5.60 mmol) was dissolved in DMF (10 mL), and NaH (0.27 g, 11.20 mmol) was added at 0°C, and the mixture was reacted for 20 minutes with stirring. Triisopropylsilyl chloride (1.60 g, 8.40 mmol, TIPSCl) was added dropwise, and the mixture was warmed to room temperature and reacted for 16 hours. The mixture was diluted with water and extracted with EA, and the organic phases were combined, dried, and concentrated. Column chromatography purification (EA / PE system) gave 1.77 g of a white solid in 95% yield.

[0181] Step 2: Synthesis of ethyl (S)-1-(5-cyano-1H-7-azaindole-4-amino)piperidine-3-carboxylate [ka]

[0182] 4-Chloro-1-(triisopropylsilyl)-1H-pyrrolo[2,3-b]pyridine-5-carbonitrile (0.90 g, 2.69 mmol) was dissolved in 1,4-dioxane (15 mL), and triethylamine (0.68 g, 6.72 mmol) and (S)-1-aminopiperidine-3-carboxylate ethyl hydrochloride (0.73 g, 3.50 mmol) were slowly added at room temperature. The mixture was heated to 100°C under nitrogen gas protection and reacted for 24 hours. The reaction solution was concentrated and purified by column chromatography (normal phase column was EA / PE system, reverse phase column was H 2 0(0.1% TFA) / MeCN system) gave 0.03 g of a pale yellow solid.

[0183] 1 H NMR (300 MHz, DMSO-d 6) δ 11.75 (br, 1H), 8.62 (br, 1H), 8.06 (s, 1H), 7.20 (br, 1H), 6.76 (br, 1H), 4.08 (q, J = 6.0 Hz, 2H), 3.21 (m, 1H), 3.05 (m, 1H), 2.81 (m, 1H), 2.66 (m, 2H), 1.99 (m, 1H), 1.75 (m, 2H), 1.32 (m, 1H), 1.17(t, J = 7.0 Hz, 3H). MS (ESI) m / z: 314 [M+H] + .

[0184] Example 22 Synthesis of (S)-4-((3-((2,2-difluoroethyl)carbamoyl)piperidin-1-yl)amino)-N-methyl-1H-pyrrolo[2,3-b]pyridine-5-formamide [ka]

[0185] (S)-4-((3-((2,2-difluoroethyl)carbamoyl)piperidin-1-yl)amino)-N-methyl-1H-pyrrolo[2,3-b]pyridine-5-formamide was synthesized by referring to the preparation methods of the compound of Example 1 and the compound of Example 2, but by replacing the aqueous ammonia solution in step 4 of Example 1 with a methylamine solution, and by replacing 4-chloro-7-azaindole-5-formamide in step 5 of Example 2 with 4-chloro-N-methyl-1H-pyrrolo[2,3-b]pyridine-5-formamide.

[0186] 1 H NMR (400 MHz, DMSO-d 6) δ 11.48 (br, 1H), 9.74 (br, 1H), 8.31 (br, 3H), 7.06 (d, J = 38.4 Hz, 2H), 5.97 (t, J = 56.0 Hz, 1H), 3.56 - 3.40 (m, 2H), 3.18 - 3.06 (m, 2H), 2.75 (d, J = 4.2 Hz, 3H), 2.68 (t, J = 10.9 Hz, 1H), 2.48 - 2.30 (m, 2H), 1.80 (d, J = 12.4 Hz, 2H), 1.73 - 1.61 (m, 1H), 1.35 (m, 1H); MS (ESI) m / z: 381 [M+H] + .

[0187] Example 23 Synthesis of (S)-ethyl 4-((3-((2,2,2-trifluoroethyl)carbamoyl)piperidin-1-yl)amino)-1H-pyrrolo[2,3-b]pyridine-5-carboxylate [ka]

[0188] The synthesis of ethyl (S)-4-((3-((2,2,2-trifluoroethyl)carbamoyl)piperidin-1-yl)amino)-1H-pyrrolo[2,3-b]pyridine-5-carboxylate was performed by referring to the preparation methods of the compounds of Example 1 and Example 22, except that the aqueous ammonia solution in Step 4 of Example 1 was replaced with a methylamine solution.

[0189] 1H NMR (400 MHz, DMSO-d6) δ 12.49 (br, 1H), 10.53 (br, 1H), 8.73 (br, 1H), 8.63 (br, 1H), 8.40 (br, 1H), 7.33 (br, 1H), 7.21 (br, 1H), 3.89 (dd, J = 10.0, 6.7 Hz, 2H), 3.11 (dd, J = 24.0, 9.1 Hz, 2H), 2.80 (s, 3H), 2.67 (m, 3H), 1.85 (m, 2H), 1.69 (m, 1H), 1.38 (m, 1H); MS (ESI) m / z: 399 [M+H]+.

[0190] Example 24 Kinase activity assay This test is 33 Using the P-ATP isotope assay, the inhibitory effect of compounds against the kinases JAK1, JAK2, JAK3, and TYK2 was tested, and the IC50 inhibitory concentration (IC) of the compounds against these kinases was calculated. 50 was obtained.

[0191] Tofacitinib (Tofacitinib) was prepared according to the method of patent WO2014195978A2 (Jiangsu Vcare PharmaTech Co. Ltd., Product No.: 321-1-1688-37C).

[0192] 1. Basic reaction buffer 20 mM 4-hydroxyethylpiperazineethanesulfonic acid buffer (Hepes, pH 7.5), 10 mM magnesium chloride (MgCl 2 ), 1 mM glycol ether diamine tetraacetic acid (EGTA), 0.02% dodecyl polyglycol ether (Brij35), 0.02 mg / ml bovine serum albumin (BSA), 0.1 mM sodium vanadate (Na 3 VO 4 ), 2 mM dithiothreitol (DTT), and 1% dimethyl sulfoxide (DMSO).

[0193] 2. Compound Preparation Compounds were dissolved in 100% DMSO to a specific concentration and then gradient diluted into samples of different concentrations (DMSO solutions) by an automated sampling device.

[0194] 3. Reaction process 3.1 Dilute the reaction substrate in the basic reaction buffer. 3.2 Add the kinase to the substrate solution and mix gently to ensure uniformity. 3.3 Different concentrations of compounds diluted in 100% DMSO are added to the kinase solution by the automated sampling system and incubated at room temperature for 20 minutes. 3.4 At room temperature 33 The kinase reaction was initiated by adding P-ATP (10 μM, 10 μCi / μl) and allowed to proceed for 2 hours.

[0195] 4. Detection The reaction mixture was filtered through an ion exchange filtration system to remove unreacted ATP and ions such as ADP generated by the reaction. 33 The radiation dose of P isotope was detected.

[0196] 5. Data Processing Based on the radiation dose, the kinase activity of the system with different concentrations of inhibitors added was calculated, and the inhibitory effects of different concentrations of compounds on the kinase activity were obtained, which were then fitted by GraphPad Prism to obtain the inhibitory IC of the compounds. 50 was calculated.

[0197] The biological activity of the compounds of the present invention was determined by the above-mentioned tests and the IC 50 The values ​​are specifically shown in Table 1.

[0198] [Table 1]

[0199] Compounds WX14 and WX15 are prepared by referring to the methods of Examples 12 and 13 of WO2016116025A1 patent, and their structures are shown below.

[0200] [ka]

[0201] Conclusion: The compounds of Example 1, Example 2, Example 3, Example 7, Example 9, and Example 23 of the present invention have better JAK1 kinase inhibitory activity than the positive control drug tofacitinib, and most of the compounds have stronger selectivity against JAK1 kinase and JAK3 kinase than JAK2, and can be expected to have better safety. The compounds of the present invention also have significant inhibitory activity against TYK2, and have corresponding anti-inflammatory activity.

[0202] Example 25 Liver microsome test The total volume of the culture system was 250μL, and a human liver microsome culture solution with a protein concentration of 0.5mg / mL was prepared in 50mmol / L PBS buffer (pH=7.4). Before starting the culture, 2.5μL of the test compound at 100μmol / L was mixed with 197.5μL of the culture solution, and the mixture was pre-cultured in a water bath at 37℃ for 5 minutes. Then, 50μL of reduced coenzyme II solution (5mmol / L) pre-cultured for 5 minutes was added to start the reaction. (The protein content of liver microsome species in the reaction system was 0.5g / L, and the final concentration of the test compound was 1μmol / L.) The mixture was cultured in a water bath at 37℃ with shaking, and the mixture was removed at 0, 5, 15, 30, and 60 minutes, and the reaction was terminated by immediately adding 600μL of a methanol solution containing a mixture of positive and negative internal standards, with terfenadine (positive ion internal standard, 25ng / mL) and tolbutamide (negative ion internal standard, 50ng / mL). After the reaction, the culture solution was shaken for 2 minutes and centrifuged for 10 minutes (4°C, 16000r / min), and the supernatant was taken and detected by LC-MS / MS to quantitatively analyze the residual amount of the parent drug (DMSO=0.1%).

[0203] The concentration of the compound incubated for 0 min was set as 100%, and the concentrations at other incubation times were converted to percentages of residual amount. Linear regression was performed on the natural logarithm of the percentage of residual amount at each time point against the incubation time to calculate the slope k, and T was calculated according to the formula. 1 / 2 The in vitro half-life was calculated as =-0.693 / k. The clearance rate in liver microsomes (CLint (μL / min / mgprotein) = Ln(2)*1000 / T 1 / 2(min) / protein concentration (mg / ml).

[0204] [Table 2]

[0205] Conclusion: The compounds of Examples 1 and 2 of the present invention have excellent hepatic metabolic stability, and in particular, the compound of Example 2 has in vivo metabolic stability with a metabolic half-life in liver microsomes that is significantly longer than that of the positive control drug tofacitinib.

[0206] Example 26 Therapeutic activity test in DSS-induced ulcerative colitis model in mice Mice aged 6-7 weeks were randomly divided into groups, with 6 mice per group. The mice in the negative control group were allowed to drink purified water ad libitum, while the mice in the other groups were allowed to drink 2% dextran sulfate sodium (DSS) solution ad libitum for 7 consecutive days. Along with establishing the model, each group was administered drug treatment once a day for 7 consecutive days (the number of treatment groups and mice can be adjusted according to the specific situation). After 8-10 days, the water containing DSS was replaced with purified water, and the mice were allowed to drink ad libitum. The treatment was then continued as usual for 3 days.

[0207] From the first day of administration, the mice were observed daily for mental status, food intake, water intake, and activity, and the mice were weighed daily. The feces of the animals were collected for evaluation of their condition and occult blood testing. The disease activity index (DAI) of the experimental animals was obtained comprehensively from 1 to 10 days.

[0208] [Table 3]

[0209] The scores were scored according to the above-mentioned standard. The scores of weight loss, fecal characteristics, and occult blood status were summed up to obtain a disease activity index (DAI) for each mouse to evaluate disease activity. DAI = (weight loss score + fecal characteristics score + fecal blood score) / 3, with a score range of 0 to 4 points.

[0210] The experimental data were statistically processed and the data were presented as mean ± SD or mean ± SME. Statistical analysis was performed using SPSS13.0 software. Differences between groups were compared using t-test (equal variance) or t'-test (unequal variance), with α = 0.05 and P < 0.05 considered to be statistically significant.

[0211] DAI evaluation results: The compounds of Example 1 (DIA score on day 8 for the 5 mg / kg group was 2.8, and for the 15 mg / kg group was 2) and Example 2 (DIA score on day 8 for the 5 mg / kg group was 1.5) showed clear therapeutic effects in the mouse DSS model, and their therapeutic effects were stronger than those of the positive control drugs mesalazine (5ASA) (DIA score on day 8 for the 100 mg / kg group was 3.5) and tofacitinib (DIA score on day 8 for the 5 mg / kg group was 3.5).

[0212] Conclusion: The compounds of Examples 1 and 2 of the present invention exhibited significant inhibitory effects on DSS-induced enteritis in mice at low doses, and their drug effects were stronger than those of the positive control drug tofacitinib at the same dose.

[0213] Example 27 A study to evaluate the therapeutic activity of DNBS (2,4-dinitrobenzene sulfonic acid) in a rat model of Crohn's disease induced by DNBS Four to five week old male Wistar rats were adaptively fed for 5 days and randomly divided into six groups 2 days before the experiment.

[0214] Blank control group: G1 (normal), four normal rats were selected and given 30% ethanol at 10 mL / kg.

[0215] Model group: 0.5 mL of 50 mg / mL DNBS (DNBS dissolved in 30% ethanol) was injected into the intestinal tract of rats to induce rat colitis. Of the total of 40 rats, 8 were selected as model group G2 (model), and the remaining 32 rats were administered the drug.

[0216] Administration groups: Rats with DNBS-induced colitis were administered ABT494 or the compound of Example 2, and divided into G3 to G6 groups of 8 rats per group: G3: ABT494 10 mg / kg QD, G4: Example 2 0.3 mg / kg BID, G5: Example 2 1 mg / kg BID, G6: Example 2 3 mg / kg BID.

[0217] The rats were fasted for 48 hours, and 5% glucose salt solution (10mL / kg) was used as an energy supplement for the experimental animals once a day during the fasting period. On the day of making the model in the experiment, the fasted rats were anesthetized with sucrose (25mg / kg), and then 0.5mL, 50mg / mL DNBS was injected from the anus of the rats to the left colon flexure (about 8cm from the anus) to induce enteritis in the rats. The rats were kept in a head-down position for 15 minutes until they regained consciousness to prevent the reflux of DNBS. Drug treatment was performed for 7 days from the day of making the model in the experiment.

[0218] The positive control drug ABT494 was purchased from Nanjing New Enzyme Pharmaceutical Technology Co Ltd, with the number NNES190329, and its structure is shown below.

[0219] [ka]

[0220] After establishing the DNBS model, the animals' weights were measured and recorded daily, and the animals' daily activities were observed and abnormal situations were recorded. During the experiment, the rats' feces were evaluated daily. On the 7th day, the animals were anesthetized with isoflurane (3–5%), blood was collected from the orbit, and serum was separated and stored at -80°C. All animals were kept in a 10-mL PBS-free room temperature for 12 h. 2The animals were euthanized by inhaling 100 mg of ethanol and then dislocating the cervical spine. The abdominal cavity was incised, the colon and rectum were removed, and dissected longitudinally to evaluate the quality of feces in the colon and rectum. After washing the colon and rectum with iced PBS, the ulcer surface was observed, the length, weight, and area of ​​the ulcer were recorded, and the ratio of the weight to the length was calculated (ratio of colon weight (g) / length (cm) × 100), and the whole was photographed. All intestinal tissues were divided longitudinally into two, fixed in 4% neutral PFA, embedded in paraffin, and the remaining colon samples were frozen and stored at -80°C.

[0221] Statistical analysis was performed using Excel and Graphpad software. Results are expressed as mean ± SD. Comparisons between groups were performed using two-tailed t-tests. * p<0.05 indicates that the drug treatment group has a significant statistical difference compared to the model group; ** p<0.01 indicates that the drug treatment group has a highly significant statistical difference compared to the model group; # p<0.05 indicates that the model group has a significant statistical difference from the normal group; ## p<0.01 indicates that the model group has a highly significant statistical difference from the normal group.

[0222] The effect of the compounds on the colon weight to length ratio in the rat DNBS-induced Crohn's disease model is shown in Figure 25, and the photographs of the colon are shown in Figures 27 to 33. Here, the colon weight to length ratios in the drug treatment groups G3: ABT494 10 mg / kg-QD, G4: Example 2 0.3 mg / kg-BID, G5: Example 2 1 mg / kg-BID, and G6: Example 2 3 mg / kg-BID were 13.4 ± 5.2, 10.5 ± 3.9, 10.0 ± 1.9, and 10.5 ± 2.0, respectively, which were all lower than the colon weight to length ratio in the model group G2 (14.6 ± 5.1), and the G5 Example 2 1 mg / kg-BID group had a significant difference compared to the model group G2 (*P<0.05).

[0223] The effect of the compounds on colonic ulcer area in a rat DNBS-induced Crohn's disease model is shown in Figure 26, where the colonic ulcer areas of mice in the G3 ABT494 10 mg / kg-QD, G4 Example 2 0.3 mg / kg-BID, G5 Example 2 1 mg / kg-BID, and G6 Example 2 3 mg / kg-BID drug treatment groups were 1.37 ± 1.72, 0.83 ± 1.00, 0.65 ± 1.05, and 0.76 ± 1.21, respectively, all of which were lower than the model group G2 (1.54 ± 2.00).

[0224] Conclusion: The compound of Example 2 of the present invention exhibits significant inhibitory effect on DNBS-induced enteritis in rats at a low dose.

Claims

1. A compound represented by formula (I), a stereoisomer thereof or a pharma- ceutically acceptable salt thereof. 【Chemistry 1】 wherein ring A optionally comprises one or more R ａ selected from the following groups substituted by 【Chemistry 2】 R １ is hydrogen, hydroxy, optionally one or more R ａ substituted by methyl, ethyl, propyl, cyclopropyl, n-butyl, t-butyl, cyclobutyl, pyridyl diphenyl, imidazolyl, pyrazolyl, oxazolyl, thiazolyl or 【Chemistry 3】 Selected from R ２ -C=ONR ６ R ７ and L is -C=ONR １０ -, -S(O) ｍ -, -C=O- or -C=OO-; Alternatively, L is absent, R ３ is hydrogen, R ４ and R ５ are hydrogen, R ６ and R ７ and R １０ are each hydrogen, optionally one or more R ｂ C substituted by １ -C ８ independently selected from alkyl, m is selected from 1 or 2; Here, R ａ is hydrogen, halogen, hydroxy, cyano, amino, C1-C8 alkyl or C １ -C ８ selected from alkoxy, R ｂ is selected from hydrogen or deuterium, If L is not present, then R １ is selected from hydrogen or hydroxy; however, 【Chemistry 4】 【Chemistry 5】 【Chemistry 6】 The present invention excludes a compound selected from the group consisting of:

2. Ring A optionally contains one or more R ａ is replaced by 【Chemistry 7】 Selected from R １ is hydrogen, hydroxy, optionally one or more R ａ methyl, ethyl, propyl, cyclopropyl, n-butyl, t-butyl, cyclobutyl, pyridinyl, imidazolyl, pyrazolyl, oxazolyl, thiazolyl or 【Chemistry 8】 Selected from R ２ -C=ONR ６ R ７ and L is -C=ONR １０ -, -S(O) ｍ -, -C=O- or -C=OO-; Alternatively, L is absent, R ３ is hydrogen, R ４ and R ５ are hydrogen, R ６ and R ７ and R １０ are each hydrogen, optionally one or more R ｂ C substituted by １ -C ８ independently selected from alkyl, m is selected from 1 or 2; Here, R ａ is hydrogen, halogen, hydroxy, cyano, amino, C １ -C ４ Alkyl or C １ -C ４ selected from alkoxy, R ｂ is selected from hydrogen or deuterium, If L is not present, then R １ is selected from hydrogen or hydroxy 2. The compound according to claim 1 .

3. Ring A optionally contains one or more R ａ is replaced by 【Chemistry 9】 Selected from R １ is hydrogen, hydroxy, optionally one or more R ａ substituted by methyl, ethyl, propyl, cyclopropyl, t-butyl, cyclobutyl or 【Chemistry 10】 Selected from R ２ -C=ONR ６ R ７ and R ６ and R ７ are hydrogen, L is -C=ONR １０ -, -S(O) ｍ -, -C=O- or -C=OO-; Alternatively, L is absent, R ３ is hydrogen, R ４ and R ５ are hydrogen, R １０ is hydrogen, m is selected from 1 or 2; Here, R ａ is selected from hydrogen, fluorine, chlorine, hydroxy, cyano, amino, methyl or methoxy; If L is not present, then R １ is selected from hydrogen or hydroxy; 3. The compound according to claim 2 .

4. A pharmaceutical composition comprising the compound according to any one of claims 1 to 3, a stereoisomer thereof or a pharma- ceutically acceptable salt thereof, and a pharma- ceutical carrier.

5. 5. The pharmaceutical composition according to claim 4, wherein the pharmaceutical composition is selected from the group consisting of capsules, powders, tablets, pellets, pills, injections, syrups, oral liquids, inhalants, ointments, suppositories, and patches.

6. The compound according to any one of claims 1 to 3, its stereoisomer or a pharma- ceutically acceptable salt thereof.

5. Use of the salt thereof or the pharmaceutical composition of claim 4 for the preparation of a medicament for preventing or treating a Janus kinase (JAK) family-induced disease.

7. 7. The use according to claim 6, characterized in that the disease is selected from immune system diseases, autoimmune diseases, skin diseases, allergic diseases, viral diseases, type 1 diabetes and diabetic complications, Alzheimer's disease, xerophthalmia, myelofibrosis, thrombocytosis, erythrocytosis or cancer.

8. 8. The use according to claim 7, characterized in that the immune system disease is organ transplant rejection, the autoimmune disease is selected from systemic lupus erythematosus, multiple sclerosis, rheumatoid arthritis, juvenile arthritis, guttate psoriasis, ulcerative colitis, Crohn's disease or autoimmune thyroid disease, the skin disease is selected from psoriasis, skin rash, alopecia areata or atopic dermatitis, the allergic disease is selected from asthma or rhinitis, the viral disease is selected from hepatitis B, hepatitis C, chicken pox or varicella zoster virus, and the cancer is selected from solid tumors, blood cancer or skin cancer.

9. The use according to claim 8, characterized in that the organ transplant rejection is selected from xenograft rejection or graft-versus-host disease, the solid tumor is selected from prostate cancer, renal cancer, liver cancer, pancreatic cancer, gastric cancer, breast cancer, lung cancer, head and neck cancer, thyroid cancer, spongioblastoma or melanoma, the blood cancer is selected from lymphoma or leukemia, and the skin cancer is selected from cutaneous T-cell lymphoma or cutaneous B-cell lymphoma.

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