Substituted tricyclic derivatives and use thereof

Abstract

Provided are a series of substituted tricyclic derivatives and the use thereof. Specifically provided are compounds represented by formula (II), (I-1) or (I-2), stereoisomers thereof and pharmaceutically acceptable salts thereof.

Description

Substituted tricyclic derivatives and uses thereof

[0001] Cross-reference to related applications

[0002] This application claims priority to the prior patent application filed with the China National Intellectual Property Office on August 8, 2024 (application number CN2024110886648) and the prior patent application filed with the China National Intellectual Property Office on January 24, 2025 (application number CN2025101225477), the contents of which are hereby incorporated by reference in their entirety. TECHNICAL FIELD

[0003] The present application relates to a series of substituted tricyclic derivatives and their uses, in particular to the compounds represented by formula (II), (I-1), (I-2), stereoisomers thereof and pharmaceutically acceptable salts thereof. BACKGROUND

[0004] Janus kinases (JAKs) are cytoplasmic tyrosine kinases, including four known family members: JAK1, JAK2, JAK3, and tyrosine kinase 2 (TYK2). JAK1, JAK2, and TYK2 are widely present in various cell tissues, and JAK3 is mainly expressed in cells of the hematopoietic system, especially myeloid and lymphoid cells. The JAK family can transmit downstream intracellular signals generated by more than 50 different immunomodulators, including cytokines, interferons, and hormones, to regulate various cell functions. When the cytokine receptor binds to type I or II cytokines, a conformational change occurs, JAK binds to it, and autophosphorylation and transphosphorylation occur, further activating the signal transducer and activator of transcription (STAT). Activated STAT enters the nucleus, regulates the transcription of various genes, and in turn regulates various cell functions, such as regulating the apoptosis, proliferation, migration, maturation, and differentiation of T cells, B cells, natural killer cells, macrophages, and epidermal cells, and also regulating the increase in the production and release of pro-inflammatory cytokines.

[0005] Different molecules of the JAK family mediate the functions of different cytokines. JAK1 and JAK3 mediate the signal transduction of IL-2, IL-4, IL-7, IL-9, IL-15 and IL-21 after binding to the type I receptor γ chain (yc) to activate downstream STAT5 or STAT6, and blocking JAK1 or JAK3 can block the effects of these cytokines. IL-6 depends on JAK1 for signal transduction. JAK2 mediates signal transduction for IL-3, IL-5, erythropoietin and granulocyte-macrophage colony-stimulating factor (GM-CSF). TYK2 mediates signal transduction of IL-12 and IL-23 to activate STAT3 or STAT4.

[0006] Studies have shown that the JAK family is involved in the occurrence of various diseases. Human JAK1 loss-of-function mutations can cause immune deficiency and pathogen infection, and high-activity mutations can cause immune dysfunction or eosinophilic syndrome. Human JAK2 high-activity mutants are associated with myelodysplasia, leukemia and lymphoma, and human loss-of-function mutations have not been reported. JAK2 deficiency (JAK2- / -) in mice can cause red blood cell production defects and death at 12-13 days after conception. Human JAK3 deficiency has been described, which presents as severe combined immunodeficiency within a few months of birth, with a lack of circulating T cells and NK cells, and abnormal B cell function, with symptoms such as developmental arrest, severe and recurrent infections, thrush and diarrhea. Human JAK3 high-activity mutations can cause leukemia. Human TYK2 low-activity can reduce the susceptibility to autoimmune diseases.

[0007] There are approved JAK inhibitors for the treatment of autoimmune diseases, including rheumatoid arthritis, psoriatic arthritis, ulcerative colitis and Crohn's disease, such as upadacitinib, abrocitinib, etc. The TYK2 selective inhibitor deucravacitinib has also been approved by the FDA. There are no JAK inhibitors approved for use in neurodegenerative diseases such as Alzheimer's disease and Parkinson's syndrome. Studies have shown that inflammation plays an important role in neurodegenerative diseases such as Alzheimer's disease, Parkinson's syndrome, multiple sclerosis and spinal cord lateral sclerosis. Therefore, the development of new JAK1 / TYK2 inhibitors has important clinical value. SUMMARY

[0008] The present application provides a compound represented by formula (II), a stereoisomer thereof or a pharmaceutically acceptable salt thereof,

[0009] wherein,

[0010] is a double bond or a single bond, and the five-membered ring in which T1, T2, T3 are located is an aromatic ring;

[0011] Ring A is selected from Ring A1 and Ring A2;

[0012] Ring A1 is an optionally substituted monocyclic 4-8 membered oxacycloalkyl; a1 substituted monocyclic 4-8 membered oxacycloalkyl;

[0013] Ring A2 is wherein 1 denotes the connection site to the imidazole N and 2 denotes the connection site to R4;

[0014] Ring B is selected from an optionally substituted monocyclic 4-8 membered nitrogen heterocycloalkyl; b substituted monocyclic 4-8 membered nitrogen heterocycloalkyl;

[0015] Ring C is annelated to Ring B, Ring C is selected from an optionally substituted 5-10 membered nitrogen containing heteroaryl; c substituted 5-10 membered nitrogen containing heteroaryl;

[0016] T is N, which is optionally quaternized or oxidized;

[0017] when Ring A is Ring A1, T1 is CR3, T2 is CR3, and T3 is O or NH;

[0018] when Ring A is Ring A2, one of T1, T2 and T3 is O, and the other two are independently selected from N and CR3;

[0019] R1 is -L1-R 11 ;

[0020] L1 is selected from a bond, -CONR 12 , -CO-, O, S, NR 12 , and the following groups optionally substituted with 1 or more R 1a : C 1-4 alkyl, C 2- 4alkenyl, C 2-4 alkynyl, C 1-4 alkoxy, C 1-4 alkylthio, C 1-4 alkylamino, C 3-8 cycloalkyl, 3-8 membered heterocycloalkyl, phenyl and 5-6 membered heteroaryl;

[0021] R 11 is selected from H, D, F, Cl, Br, I, CN, and the following groups optionally substituted with 1 or more R 1a : C 1-4 alkyl, C 2-4 alkenyl, C 2-4 alkynyl, C 1-4 alkoxy, C1-4 Alkylthio, C 1-4 Alkylamino, -C 3-8 Cycloalkyl, 3-8 membered heterocycloalkyl, phenyl and 5-6 membered heteroaryl;

[0022] R 12 Selected from H and arbitrarily selected by one or more R 1a The following groups are substituted: C 1-4 Alkyl, C 2-4 alkenyl, C 2-4 alkynyl group, C 3-8 Cycloalkyl and 3-8 membered heterocyclic alkyl groups;

[0023] R2 is selected from H, D, F, Cl, Br, I, OH, NH2, CN, and optionally one or more R 2a The following groups are substituted: C 1-4 Alkyl, C 2-4 alkenyl, C 2-4 alkynyl group, C 1-4 Alkoxy, C 1-4 Alkylthio, C 1-4 Alkylamino, C 3-8 Cycloalkyl, 3-8 membered heterocycloalkyl, phenyl and 5-6 membered heteroaryl;

[0024] R3 is selected from H, D, F, Cl, Br, I, OH, NH2, CN, and optionally one or more R 3a The following groups are substituted: C 1-4 Alkyl, C 2-4 alkenyl, C 2-4 alkynyl group, C 1-4 Alkoxy, C 1-4 Alkylthio, C 1-4 Alkylamino, C 3-8 Cycloalkyl, 3-8 membered heterocycloalkyl, phenyl and 5-6 membered heteroaryl;

[0025] Alternatively, two R3 atoms on adjacent atoms can be connected to form a structure optionally bounded by one or more R atoms. 3b The following groups are substituted: C 5-8 Cycloalkyl, 5-8 membered heterocycloalkyl or 5-6 membered heteroaryl;

[0026] R4 is selected from CN, C 1-4 Alkyl, C 2-4 alkenyl, C 2-4 alkynyl group, C 1-4 Alkoxy, C 1-4 Alkylthio, C 1-4 Alkylamino, C 3- 8-membered cycloalkyl, 3-8-membered heterocycloalkyl, phenyl and 5-6-membered heteroaryl, wherein C 1-4 Alkyl, C2-4 alkenyl, C 2-4 alkynyl, C 1-4 alkoxy, C 1- alkylthio, C 3-8 cycloalkyl, 3-8 membered heterocycloalkyl, phenyl and 5-6 membered heteroaryl, which are substituted with CN, and further optionally substituted with one or more R 4a substituents;

[0027] each R a1 , each R b , each R c , each R 1a , R 11a , R 11b , R 11c , R 11d , each R 2a , each R 3a , each R 3b , each R 4a are each independently selected from the group consisting of H, D, F, CI, Br, I, OH, NH2, CN, =0 and the following groups, which are optionally substituted with one or more R 1-4 alkyl, C 2-4 alkenyl, C 2-4 alkynyl, C 1-4 alkoxy, C 1-4 alkylthio, C 3-8 cycloalkyl, 3-8 membered heterocycloalkyl, phenyl and 5-6 membered heteroaryl;

[0028] or 2 R a1 are linked together, or 2 R b are linked together, or 2 R c are linked together, or R b and R c are linked together, are each independently forming the following groups, which are optionally substituted with one or more R 3-8 cycloalkyl, 4-8 membered heterocycloalkyl, phenyl or 5-6 membered heteroaryl;

[0029] each R is independently selected from the group consisting of H, D, F, CI, Br, I, OH, NH2, CN and the following groups, which are optionally substituted with one or more F: 1-4 alkyl, C 2-4 alkenyl, C 2-4 alkynyl, C 1-4 alkoxy, C 1-4 alkylthio, C 1-4 alkylamino, C 3-8 cycloalkyl, 3-8 membered heterocycloalkyl, phenyl and 5-6 membered heteroaryl.

[0030] The application also provides a compound represented by formula (I), a stereoisomer thereof or a pharmaceutically acceptable salt thereof,

[0031] wherein,

[0032] is a double bond or a single bond, and the five-membered ring in which T1, T2, T3 are located is an aromatic ring;

[0033] ring A is selected from ring A1 and ring A2;

[0034] ring A1 is a monocyclic 4-8 membered oxacycloalkyl group optionally substituted with 1 or more R a1 ;

[0035] ring A2 is wherein 1 represents the connection site with the imidazole N, and 2 represents the connection site with R4;

[0036] ring B is selected from a monocyclic 4-8 membered azacycloalkyl group optionally substituted with 1 or more R b ;

[0037] ring C is annelated with ring B, and ring C is selected from a 5-10 membered nitrogen-containing heteroaryl group optionally substituted with 1 or more R c ;

[0038] when ring A is ring A1, T1 is CR3, T2 is CR3, and T3 is O or NH;

[0039] when ring A is ring A2, one of T1, T2 and T3 is O, and the other two are independently selected from N and CR3;

[0040] R1 is -L1-R 11 ;

[0041] L1 is selected from a bond, -CONR 12 , -CO-, O, S, NR 12 , and the following group optionally substituted with 1 or more R 1a : C 1-4 alkyl, C 2- 4alkenyl, C 2-4 alkynyl, C 1-4 alkoxy, C 1-4 alkylthio, C 1-4 alkylamino, C 3-8 cycloalkyl, 3-8 membered heterocycloalkyl, phenyl and 5-6 membered heteroaryl;

[0042] R 11 is selected from H, F, Cl, Br, I, CN and the following group optionally substituted with 1 or more R 1a : C 1-4 alkyl, C 2-4alkenyl, C 2- 4alkynyl, C 1-4 alkoxy, C 1-4 alkylthio, C 1-4 alkylamino, C 3-8 cycloalkyl, 3-8 membered heterocycloalkyl, phenyl and 5-6 membered heteroaryl;

[0043] R 12 selected from H and the following groups optionally substituted with 1 or more R 1a substituted: C 1-4 alkyl, C 2-4 alkenyl, C 2-4 alkynyl, C 3-8 cycloalkyl and 3-8 membered heterocycloalkyl;

[0044] R2is selected from H, F, Cl, Br, I, OH, NH2, CN and the following groups optionally substituted with 1 or more R 2a substituted: C 1-4 alkyl, C 2-4 alkenyl, C 2-4 alkynyl, C 1-4 alkoxy, C 1-4 alkylthio, C 1-4 alkylamino, C 3-8 cycloalkyl, 3-8 membered heterocycloalkyl, phenyl and 5-6 membered heteroaryl;

[0045] R3is selected from H, F, Cl, Br, I, OH, NH2, CN and the following groups optionally substituted with 1 or more R 3a substituted: C 1-4 alkyl, C 2-4 alkenyl, C 2-4 alkynyl, C 1-4 alkoxy, C 1-4 alkylthio, C 1-4 alkylamino, C 3-8 cycloalkyl, 3-8 membered heterocycloalkyl, phenyl and 5-6 membered heteroaryl;

[0046] or 2 R3on adjacent atoms are joined to form a group optionally substituted with 1 or more R 3b substituted: C 5-8 cycloalkyl, 5-8 membered heterocycloalkyl or 5-6 membered heteroaryl;

[0047] R4is selected from CN, C 1-4 alkyl, C 2-4 alkenyl, C 2-4 alkynyl, C 1-4 alkoxy, C 1-4 alkylthio, C 1-4 alkylamino, C 3-8 cycloalkyl, 3-8 membered heterocycloalkyl, phenyl and 5-6 membered heteroaryl, which C 1-4 alkyl, C 2-4 alkenyl, C 2-4 alkynyl, C 1-4 alkoxy, C 1- alkylthio, C 3-8 cycloalkyl, 3-8 membered heterocycloalkyl, phenyl and 5-6 membered heteroaryl, which C 4a is further optionally substituted with 1 or more R a1 ;

[0048] each R b , each R c , each R 1a , each R 2a , each R 3a , each R 3b , each R 4a is independently selected from the group consisting of H, F, Cl, Br, I, OH, NH2, CN, =0 and the following groups, which are optionally substituted with 1 or more R 1-4 alkyl, C 2-4 alkenyl, C 2-4 alkynyl, C 1-4 alkoxy, C 1-4 alkylthio, C 3-8 cycloalkyl, 3-8 membered heterocycloalkyl, phenyl and 5-6 membered heteroaryl;

[0049] or 2 R a1 are linked together, or 2 R b are linked together, or 2 R c are linked together, or R b and R c are linked together, are independently forming the following groups, which are optionally substituted with 1 or more R 3-8 cycloalkyl, 4-8 membered heterocycloalkyl, phenyl or 5-6 membered heteroaryl;

[0050] each R is independently selected from the group consisting of H, F, Cl, Br, I, OH, NH2, CN and the following groups, which are optionally substituted with 1 or more F: C 1-4 alkyl, C 2-4 alkenyl, C 2-4 alkynyl, C 1-4 alkoxy, C 1-4 alkylthio, C 1-4 alkylamino, C 3-8 cycloalkyl, 3-8 membered heterocycloalkyl, phenyl and 5-6 membered heteroaryl.

[0051] In some embodiments of the application, each R is independently selected from H, D, F, Cl, OH, NH2, CN, CH3, and CF3, and the other variables are as defined in the application.

[0052] In some embodiments of the application, each R is independently selected from H, F, Cl, OH, NH2, CN, CH3, and CF3, and the other variables are as defined in the application.

[0053] In some embodiments of the application, each R is independently selected from F, and the other variables are as defined in the application.

[0054] In some embodiments of the application, each R a1 is independently selected from H, D, F, Cl, Br, I, OH, NH2, CN, and CH3, CH2CH3, vinyl, ethynyl, OCH3, OCH2CH3, cyclopropyl, and cyclobutyl optionally substituted with 1 or more R, and the other variables are as defined in the application.

[0055] In some embodiments of the application, each R a1 is independently selected from H, F, Cl, Br, I, OH, NH2, CN, and CH3, CH2CH3, vinyl, ethynyl, OCH3, OCH2CH3, cyclopropyl, and cyclobutyl optionally substituted with 1 or more R, and the other variables are as defined in the application.

[0056] In some embodiments of the application, each R a1 is independently selected from H, D, F, Cl, CH3, and CF3, and the other variables are as defined in the application.

[0057] In some embodiments of the application, each R a1 is independently selected from H, F, Cl, CH3, and CF3, and the other variables are as defined in the application.

[0058] In some embodiments of the application, the 2 R a1 are joined together to form a cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, oxetanyl, or oxolanyl group optionally substituted with 1 or more R, and the other variables are as defined in the application.

[0059] In some embodiments of the application, the 2 R a1 are joined together to form a cyclopropyl or cyclobutyl group optionally substituted with 1 or more R, and the other variables are as defined in the application.

[0060] In some embodiments of the application, each R bThe groups are independently selected from H, D, F, Cl, Br, I, OH, NH2, CN, =O, and the following groups optionally substituted with one or more R: CH3, CH2CH3, CH2CH2CH3, CH(CH3)2, vinyl, propenyl, ethynyl, propynyl, OCH3, OCH2CH3, OCH(CH3)2, SCH3, SCH2CH3, cyclopropyl, and cyclobutyl, with other variables as defined in this invention.

[0061] In some technical solutions of the present invention, the above-mentioned R b The variables are independently selected from H, D, F, Cl, CH3 and CF3, respectively, and other variables are as defined in this invention.

[0062] In some technical solutions of the present invention, the above-mentioned R b The groups are independently selected from H, F, Cl, Br, I, OH, NH2, CN, =O, and the following groups optionally substituted with one or more R: CH3, CH2CH3, CH2CH2CH3, CH(CH3)2, vinyl, propenyl, ethynyl, propynyl, OCH3, OCH2CH3, OCH(CH3)2, SCH3, SCH2CH3, cyclopropyl, and cyclobutyl, with other variables as defined in this invention.

[0063] In some technical solutions of the present invention, the above-mentioned R b The variables are independently selected from H, F, Cl, CH3 and CF3, respectively, and other variables are as defined in this invention.

[0064] In some technical solutions of the present invention, the above two Rs b They are linked together to form cyclopropyl or cyclobutyl groups, which may be optionally substituted with one or more R groups, and other variables are as defined in this invention.

[0065] In some technical solutions of the present invention, the above-mentioned R c The groups are independently selected from H, D, F, Cl, Br, I, OH, NH2, CN, =O, and the following groups optionally substituted with one or more R: CH3, CH2CH3, CH2CH2CH3, CH(CH3)2, vinyl, propenyl, ethynyl, propynyl, OCH3, OCH2CH3, OCH(CH3)2, SCH3, SCH2CH3, cyclopropyl, and cyclobutyl, with other variables as defined in this invention.

[0066] In some technical solutions of the present invention, the above-mentioned R c The variables are independently selected from H, D, F, Cl, CH3 and CF3, respectively, and other variables are as defined in this invention.

[0067] In some technical solutions of the present invention, the above-mentioned R ceach R is independently selected from H, F, Cl, Br, I, OH, NH2, CN, =0, and the following groups optionally substituted with 1 or more R: CH3, CH2CH3, CH2CH2CH3, CH(CH3)2, vinyl, propenyl, ethynyl, propynyl, OCH3, OCH2CH3, OCH(CH3)2, SCH3, SCH2CH3, cyclopropyl, and cyclobutyl, with the other variables as defined herein.

[0068] In some embodiments of the application, each R c is independently selected from H, F, Cl, CH3, and CF3, with the other variables as defined herein.

[0069] In some embodiments of the application, each R 1a , R 11a , R 11b , R 11c , R 11d , each R 2a , each R 3a , each R 3b , each R 4a is independently selected from H, D, F, Cl, Br, I, OH, NH2, CN, =0, and the following groups optionally substituted with 1 or more R: CH3, CH2CH3, CH2CH2CH3, CH(CH3)2, vinyl, propenyl, ethynyl, propynyl, OCH3, OCH2CH3, OCH(CH3)2, SCH3, SCH2CH3, cyclopropyl, and cyclobutyl, with the other variables as defined herein.

[0070] In some embodiments of the application, each R 1a , R 11a , R 11b , R 11c , R 11d , each R 2a , each R 3a , each R 3b , each R 4a is independently selected from H, D, F, Cl, CN, =0, CH3, CFH2, CF2H, CF3, OCH3, OCF3, and cyclopropyl, with the other variables as defined herein.

[0071] In some embodiments of the application, each R 1a , each R 2a , each R 3a , each R 3b , each R 4aeach R is independently selected from H, F, Cl, Br, I, OH, NH2, CN, =0, and the following groups optionally substituted with 1 or more R: CH3, CH2CH3, CH2CH2CH3, CH(CH3)2, vinyl, propenyl, ethynyl, propynyl, OCH3, OCH2CH3, OCH(CH3)2, SCH3, SCH2CH3, cyclopropyl, and cyclobutyl, with the other variables as defined herein.

[0072] In some embodiments of the application, each R 1a is independently selected from H, F, Cl, CN, =0, CH3, CFH2, CF2H, CF3, OCH3, OCF3, and cyclopropyl, with the other variables as defined herein. 2a 3a 3b 4a In some embodiments of the application, each R 1a is independently selected from H, F, Cl, CN, =0, CH3, CFH2, CF2H, CF3, OCH3, OCF3, and cyclopropyl, with the other variables as defined herein.

[0073] In some embodiments of the application, L1is selected from a bond, C(O)NH, C(O), O, S, NH, and the following groups optionally substituted with 1 or more R 1a : CH2, CH2CH2, vinyl, ethynyl, propynyl, OCH2, OCH2CH2, and cyclopropyl, with the other variables as defined herein.

[0074] In some embodiments of the application, L1is selected from a bond, C(O), O, S, and the following groups optionally substituted with 1 or more R 1a : C(O)NH, NH, CH2, CH2CH2, vinyl, ethynyl, propynyl, OCH2, OCH2CH2, and cyclopropyl, with the other variables as defined herein.

[0075] In some embodiments of the application, L1is selected from a bond, O, S, and the following groups optionally substituted with 1 or more F or D: CH2, CH2CH2, OCH2, OCH2CH2, and cyclopropyl.

[0076] In some embodiments of the application, each R 11 is selected from H, D, F, Cl, Br, I, CN, and the following groups optionally substituted with 1 or more R 1a : CH3, CH2CH3, CH2CH2CH3, CH(CH3)2, vinyl, ethynyl, propynyl, OCH3, OCH2CH3, OCH(CH3)2, SCH3, SCH2CH3, NHCH3, NHCH2CH3, N(CH3)2, cyclopropyl, cyclobutyl, oxetanyl, oxetanyl, with the other variables as defined herein.

[0077] ​In some technical solutions of the present invention, the above-mentioned R 11 Selected from H, F, Cl, Br, I, CN and optionally one or more R 1a The following groups may be substituted: CH3, CH2CH3, CH2CH2CH3, CH(CH3)2, vinyl, ethynyl, propynyl, OCH3, OCH2CH3, OCH(CH3)2, SCH3, SCH2CH3, NHCH3, NHCH2CH3, N(CH3)2, cyclopropyl, cyclobutyl, oxecyclobutyl, and oxecyclopentyl, with other variables as defined in this invention.

[0078] In some technical solutions of the present invention, the above-mentioned R 11 The following groups are selected from H, D, F, Cl, Br, I, CN, and optionally substituted with one or more F or D: CH3, CH2CH3, CH2CH2CH3, CH(CH3)2, OCH3, OCH2CH3, OCH(CH3)2, cyclopropyl, cyclobutyl, oxacyclobutyl, and oxacyclopentyl, with other variables as defined in this invention. In some technical solutions of this invention, R1 is selected from H, D, F, Cl, Br, I, CN, CH3, CFH2, CF2H, CF3, CD3, CH2CH3, CH(CH3)2, CH2CN, CH2CH2CN, -C(O)CH3, -C(O)CH=CH2, OCH3, OCF3, OCD3, OCH2CH3, OCH(CH3)2, SCH3, SCF3, SCH2CH3, Other variables are as defined in this invention.

[0079] In some technical solutions of the present invention, R1 is selected from H, F, Cl, Br, I, CN, CH3, CFH2, CF2H, CF3, CH2CH3, CH(CH3)2, CH2CN, CH2CH2CN, -C(O)CH3, -C(O)CH=CH2, OCH3, OCF3, OCH2CH3, OCH(CH3)2, SCH3, SCF3, SCH2CH3, Other variables are as defined in this invention.

[0080] In some technical solutions of the present invention, R1 is selected from H, D, F, Cl, Br, I, CN, and optionally one or more R. 1a The following groups are substituted: C 1-4 Alkyl, -OC 1-4 Alkyl and -SC 1-4 Alkyl groups, and other variables as defined in this invention.

[0081] In some embodiments of the application, R1is selected from H, D, F, CI, Br, I, CN, CH3, CFH2, CF2H, CF3, CD3, CH2CH3, CH(CH3)2, CH2CN, CH2CH2CN, OCH3, OCF3, OCD3, OCH2CH3, OCH(CH3)2, SCH3, SCF3, SCH2CH3, and other variables are as defined in the application.

[0082] In some embodiments of the application, L1is selected from a bond, O, and S, and R 11 is selected from F, CI, Br, I, CN, and the following groups optionally substituted with 1 or more R 1a substituents: C 1-4 alkyl, C 3-6 cycloalkyl, and 3-6 membered heterocycloalkyl, and other variables are as defined in the application.

[0083] In some embodiments of the application, R1is selected from CN, CH3, CFH2, CF2H, CF3, CD3, OCH3, OCF3, OCD3, and other variables are as defined in the application.

[0084] In some embodiments of the application, R1is selected from and other variables are as defined in the application.

[0085] In some embodiments of the application, R2is selected from H, D, F, CI, Br, I, OH, NH2, CN, and the following groups optionally substituted with 1 or more R 2a substituents: CH3, CH2CH3, CH2CH2CH3, CH(CH3)2, vinyl, ethynyl, propynyl, OCH3, OCH2CH3, OCH(CH3)2, SCH3, SCH2CH3, and cyclopropyl, and other variables are as defined in the application.

[0086] In some embodiments of the application, R2is selected from H, D, F, CI, Br, CN, CH3, CF3, ethynyl, and cyclopropyl, and other variables are as defined in the application.

[0087] In some embodiments of the application, R2is selected from H, F, CI, Br, I, OH, NH2, CN, and the following groups optionally substituted with 1 or more R 2a substituents: CH3, CH2CH3, CH2CH2CH3, CH(CH3)2, vinyl, ethynyl, propynyl, OCH3, OCH2CH3, OCH(CH3)2, SCH3, SCH2CH3, and cyclopropyl, and other variables are as defined in the application.

[0088] In some embodiments of the application, R2is selected from H, F, CI, Br, CN, CH3, CF3, ethynyl, and cyclopropyl, and the other variables are as defined in the Summary.

[0089] In some embodiments of the application, R2is selected from H, F, CI, CH3, and CF3, and the other variables are as defined in the Summary.

[0090] In some embodiments of the application, R2is H, and the other variables are as defined in the Summary.

[0091] In some embodiments of the application, R3is selected from H, D, F, CI, Br, I, OH, NH2, CN, and the following groups optionally substituted with 1 or more R 3a substituted with 1 or more R

[0092] In some embodiments of the application, R3is selected from H, D, F, CI, Br, CN, CH3, CF3, ethynyl, and cyclopropyl, and the other variables are as defined in the Summary.

[0093] In some embodiments of the application, R3is selected from H, F, CI, Br, I, OH, NH2, CN, and the following groups optionally substituted with 1 or more R 3a substituted with 1 or more R

[0094] In some embodiments of the application, R3is selected from H, F, CI, Br, CN, CH3, CF3, ethynyl, and cyclopropyl, and the other variables are as defined in the Summary.

[0095] In some embodiments of the application, R3is H, and the other variables are as defined in the Summary.

[0096] In some embodiments of the application, R4is selected from CN, CH3, CH2CH3, CH2CH2CH3, CH(CH3)2, vinyl, propenyl, ethynyl, propynyl, OCH3, OCH2CH3, OCH(CH3)2, SCH3, SCH2CH3, cyclopropyl, and cyclobutyl, said CH3, CH2CH3, CH2CH2CH3, CH(CH3)2, vinyl, propenyl, ethynyl, propynyl, OCH3, OCH2CH3, OCH(CH3)2, SCH3, SCH2CH3, cyclopropyl, and cyclobutyl are further optionally substituted with 1 or more R 4a and the other variables are as defined in the application.

[0097] In some embodiments of the application, R4is selected from CN, -CH2CN, -C(CH3)2CN, and the other variables are as defined in the application.

[0098] In some embodiments of the application, ring A1is selected from oxetanyl, oxetanyl optionally substituted with 1 or more R a1 and the other variables are as defined in the application.

[0099] In some embodiments of the application, ring A1is selected from and the other variables are as defined in the application.

[0100] In some embodiments of the application, ring B is selected from pyrrolidinyl, piperidinyl, homopiperidinyl, optionally substituted with 1 or more R b and ring C is selected from imidazolyl, pyrazolyl, pyrrolyl, pyridazinyl, and pyrimidinyl, optionally substituted with 1 or more R c and the other variables are as defined in the application.

[0101] In some embodiments of the application, ring A2is selected from and the other variables are as defined in the application.

[0102] In some embodiments of the application, ring A2is selected from and the other variables are as defined in the application.

[0103] In some embodiments of the application, ring A2is selected from and the other variables are as defined in the application.

[0104] In some embodiments of the present application, the structural unit is selected from The other variables are as defined in the present application.

[0105] In some embodiments of the present application, the structural unit is selected from The other variables are as defined in the present application.

[0106] In some embodiments of the present application, the structural unit is selected from The other variables are as defined in the present application.

[0107] In some embodiments of the present application, the structural unit is ; The other variables are as defined in the present application.

[0108] In some embodiments of the present application, the compound of formula (I) or (II), a stereoisomer thereof, or a pharmaceutically acceptable salt thereof is selected from:

[0109] wherein,

[0110] m and n are each independently selected from 0, 1, 2 and 3;

[0111] R1, R2, R3, R4, R a1 , ring B, ring C are as defined in the present application.

[0112] In some embodiments of the present application, the compound of formula (I) or (II), a stereoisomer thereof, or a pharmaceutically acceptable salt thereof is selected from:

[0113] wherein,

[0114] m and n are each independently selected from 0, 1, 2 and 3;

[0115] R1is -L1-R 11 ;

[0116] L1is selected from a bond, -CONH-, -CO-, O, S, NH and the following groups optionally substituted with 1 or more R 1a ; C 1-4 alkyl, C 2-4 alkenyl, C 2-4 alkynyl, C 1-4 alkoxy, C 1-4 alkylthio, C 1-4 alkylamino, C 3-8 cycloalkyl and 3-8 membered heterocycloalkyl;

[0117] R 11 H, F, Cl, Br, I, CN and optionally substituted by 1 or more R 1a C 1-4 alkyl, C 2-4 alkenyl, C 2- alkynyl, C 1-4 alkoxy, C 3-8 cycloalkyl and 3-8 membered heterocycloalkyl;

[0118] R2is selected from H, F, Cl, Br, I, OH, NH2, CN and optionally substituted by 1 or more R 2a C 1-4 alkyl, C 2-4 alkenyl, C 2-4 alkynyl, C 1-4 alkoxy, C 3-8 cycloalkyl and 3-8 membered heterocycloalkyl;

[0119] R3is selected from H, F, Cl, Br, I, OH, NH2, CN and optionally substituted by 1 or more R 3a C 1-4 alkyl, C 2-4 alkenyl, C 2-4 alkynyl, C 1-4 alkoxy, C 3-8 cycloalkyl and 3-8 membered heterocycloalkyl;

[0120] R4is selected from CN, CH3, CH2CH3, CH2CH2CH3, CH(CH3)2, ethenyl, propenyl, ethynyl, propynyl, OCH3, OCH2CH3, OCH(CH3)2, SCH3, SCH2CH3, cyclopropyl and cyclobutyl, which CH3, CH2CH3, CH2CH2CH3, CH(CH3)2, ethenyl, propenyl, ethynyl, propynyl, OCH3, OCH2CH3, OCH(CH3)2, SCH3, SCH2CH3, cyclopropyl and cyclobutyl are further optionally substituted by 1 or more R 4a ;

[0121] Ring B is selected from monocyclic 5-7 membered nitrogen heterocycloalkyl optionally substituted by 1 or more R b ;

[0122] Ring C is fused to Ring B, Ring C is selected from 5-6 membered nitrogen containing heteroaryl optionally substituted by 1 or more R c ;

[0123] R a1 , R 2a , R 3a , R 4aR b R c The following groups, selected independently from H, F, Cl, Br, I, OH, NH2, CN, =O, and optionally substituted with one or more F groups: C 1-4 Alkyl, C 2-4 alkenyl, C 2-4 alkynyl group, C 1-4 Alkoxy, C 1-4 Alkylthio, C 3-6 Cycloalkyl, 3-6 membered heterocyclic alkyl.

[0124] In some technical solutions of the present invention, the compounds of formula (I) or formula (II), their stereoisomers, or pharmaceutically acceptable salts thereof are selected from:

[0125] in,

[0126] m is selected from 0, 1, 2, and 3;

[0127] n is selected from 1, 2, and 3;

[0128] R1 is selected from F, Cl, Br, I, CN, and optionally one or more R 1a The following groups are substituted: C 1-4 Alkyl, -OC 1-4 Alkyl and -SC 1-4 alkyl;

[0129] R2 is selected from H, F, Cl, Br, I, and CH3;

[0130] Each R3 is independently selected from H, F, Cl, Br, I, and CH3;

[0131] R4 is selected from CN, CH2CN, CH2CH2CN, CH(CH3)CN, C(CH3)2CN,

[0132] Each R 1a They were each independently selected from H, D, F, Cl, and CN;

[0133] Each R a1 The C atoms are independently selected from H, F, Cl, Br, I, CN, =O, and C atoms optionally substituted by one or more F atoms. 1-4 alkyl.

[0134] In some technical solutions of the present application, the compound of formula (I-1), the stereoisomer thereof or the pharmaceutically acceptable salt thereof is selected from the compound of formula (I-1-1), (I-1-1a) or (I-1-1b), the compound of formula (I-2), the stereoisomer thereof or the pharmaceutically acceptable salt thereof is selected from the compound of formula (I-2-1), (I-2-1a) or (I-2-1b), and the compound of formula (I-3), the stereoisomer thereof or the pharmaceutically acceptable salt thereof is selected from the compound of formula (I-3-1), (I-3-1a) or (I-3-1b):

[0135] wherein,

[0136] m and s are each independently selected from 0, 1, 2 and 3;

[0137] R1, R2, R3, R a1 , R b , R c as defined in the present application.

[0138] In some technical solutions of the present application, the compound of formula (I-1), (I-1-1), (I-1-1a), (I-1-1b), (I-1-1c), (I-1-1d), the stereoisomer thereof or the pharmaceutically acceptable salt thereof is selected from:

[0139] wherein, R1 is as defined in the present application.

[0140] In some technical solutions of the present application, the compound of formula (I-1), (I-1-1), (I-1-1a), (I-1-1b), (I-1-1c), (I-1-1d), the stereoisomer thereof or the pharmaceutically acceptable salt thereof is selected from:

[0141] wherein,

[0142] R1 is -L1-R 11 ;

[0143] L1 is selected from a bond, O and S;

[0144] R 11 is selected from F, Cl, Br, I, CN and the following groups optionally substituted with 1 or more R 1a C 1-4 alkyl, C 3-6 cycloalkyl and 3-6 membered heterocycloalkyl;

[0145] each R 1aThe compounds are independently selected from H, D, F, Cl, CN, =O, CH3, CFH2, CF2H, CF3, OCH3, OCF3, and cyclopropyl. In some embodiments of the present invention, the compounds shown in (II-1), (II-1a), (II-1b), (II-1c), or (II-1d) above, their stereoisomers, or pharmaceutically acceptable salts thereof, wherein L1 is selected from the bonds and O, R... 11 Selected from one or more R 1a The following groups are substituted: C 1-3 Alkyl, cyclopropyl, cyclobutyl, and oxecyclobutyl, and other variables as defined in this invention.

[0146] In some technical solutions of the present invention, the compounds shown in formulas (I-1), (I-1-1), (I-1-1a), (I-1-1b), (I-1-1c), and (I-1-1d), their stereoisomers, or pharmaceutically acceptable salts thereof are selected from:

[0147] in,

[0148] R1 is selected from F, Cl, Br, I, CN, and optionally one or more R 1a The following groups are substituted: C 1-4 Alkyl, -OC 1-4 Alkyl and -SC 1-4 alkyl;

[0149] Each R 1a They were each independently selected from H, D, F, Cl, and CN;

[0150] R4 is selected from CN, CH2CN, CH2CH2CN, CH(CH3)CN, C(CH3)2CN,

[0151] In some technical solutions of the present invention, the compounds shown in (II-1), (II-1a), (II-1b), (II-1c) or (II-1d) above, their stereoisomers or pharmaceutically acceptable salts thereof, wherein R 1a Selected from F and D, other variables are as defined in this invention.

[0152] In some technical solutions of the present invention, the compounds of formula (I) or formula (I-1), their stereoisomers, or pharmaceutically acceptable salts thereof are selected from:

[0153] in,

[0154] R1 is selected from F, Cl, Br, I, CN, and optionally one or more R 1a The following groups are substituted: C1-4 alkyl, -O-C 1-4 alkyl and -S-C 1-4 alkyl;

[0155] each R 1a is independently selected from H, D, F, Cl and CN;

[0156] R4is selected from CN, CH2CN, CH2CH2CN, CH(CH3)CN, C(CH3)2CN,

[0157] In some embodiments of the application, the compound of formula (I), (I-1), (II-1), (II-1a), (II-1b), (II-1c), (II-1d), (III-1), (III-1a), (III-1b), (III-1c), (III-1d), a stereoisomer thereof, or a pharmaceutically acceptable salt thereof is wherein R1is selected from F, Cl, Br, I, CN, and CH3or OCH3optionally substituted with 1, 2, or 3 F or D.

[0158] In some embodiments of the application, the compound of formula (I), (I-1), (II-1), (II-1a), (II-1b), (II-1c), (II-1d), (III-1), (III-1a), (III-1b), (III-1c), (III-1d), a stereoisomer thereof, or a pharmaceutically acceptable salt thereof is wherein R4is selected from CH2CN, and the other variables are as defined in the application.

[0159] In some embodiments of the application, the compound of formula (II), a stereoisomer thereof, or a pharmaceutically acceptable salt thereof is selected from:

[0160] wherein ring A, L1, T, T1, T2, T3, R 11a , R 11b , R 11c , R4and are as defined in the application.

[0161] In some embodiments of the application, the compound of formula (II), a stereoisomer thereof, or a pharmaceutically acceptable salt thereof is selected from:

[0162] wherein,

[0163] L1is selected from a bond, O, S, and CH2, CH2CH2, OCH2, OCH2CH2, and cyclopropyl optionally substituted with 1 or more F or D.

[0164] R 11a R 11b R 11c R 11d are each independently selected from the group consisting of H, D, F, Cl, Br, I, OH, NH2, CN, and C 1-3 alkyl, C 1-3 alkoxy, C 1-3 alkylthio, C 3-6 cycloalkyl, and 3-6 membered heterocycloalkyl;

[0165] each R a1 is independently selected from the group consisting of H, D, F, Cl, CH3, and CF3;

[0166] R2is selected from the group consisting of H, D, F, Cl, Br, CN, CH3, CF3, ethynyl, and cyclopropyl;

[0167] R3is selected from the group consisting of H, D, F, Cl, Br, CN, CH3, CF3, ethynyl, and cyclopropyl;

[0168] R4is selected from the group consisting of CN, C 1-3 alkyl and C 3-6 cycloalkyl, said C 1-3 alkyl and C 3-6 cycloalkyl are each independently optionally substituted with CN, further optionally substituted with 1 or more F or D;

[0169] Ring B, Ring C are as defined in the present application.

[0170] Some embodiments of the present application have any combination of the above variables.

[0171] The present application also provides the compounds shown in Table A, stereoisomers thereof, or pharmaceutically acceptable salts thereof.

[0172] In some embodiments of the present application, the compounds of Table A, stereoisomers thereof, or pharmaceutically acceptable salts thereof are selected from the compounds of Table A1.

[0173] Table A compounds

[0174] Table A1 compounds

[0175] The present application also provides the A crystalline form of compound 3,

[0176] wherein said Form A has a Cu Kα radiation X-ray powder diffraction pattern comprising characteristic diffraction peaks at the following 2θ angles: 9.01 ± 0.20°, 13.27 ± 0.20° and 22.37 ± 0.20°.

[0177] In some embodiments of the application, the Cu Kα radiation X-ray powder diffraction pattern of the Form A described above, expressed in terms of 2θ angles, comprises at least 5, 6, 7 or 8 diffraction peaks selected from the group consisting of: 9.01 ± 0.20°, 13.27 ± 0.20°, 16.85 ± 0.20°, 17.91 ± 0.20°, 18.64 ± 0.20°, 20.57 ± 0.20°, 22.37 ± 0.20° and 24.54 ± 0.20°.

[0178] In some embodiments of the application, the Cu Kα radiation X-ray powder diffraction pattern of the Form A described above has characteristic diffraction peaks at the following 2θ angles: 9.01 ± 0.20°, 13.27 ± 0.20°, 17.91 ± 0.20°, 18.64 ± 0.20° and 22.37 ± 0.20°.

[0179] In some embodiments of the application, the Cu Kα radiation X-ray powder diffraction pattern of the Form A described above has characteristic diffraction peaks at the following 2θ angles: 9.01 ± 0.20°, 13.27 ± 0.20°, 16.85 ± 0.20°, 17.91 ± 0.20°, 18.64 ± 0.20°, 20.57 ± 0.20°, 22.37 ± 0.20° and 24.54 ± 0.20°.

[0180] In some embodiments of the application, the Cu Kα radiation X-ray powder diffraction pattern of the Form A described above, expressed in terms of 2θ angles, comprises at least 10, 11, 12, or 13 diffraction peaks selected from the group consisting of: 9.01 ± 0.20°, 10.62 ± 0.20°, 12.82 ± 0.20°, 13.27 ± 0.20°, 16.85 ± 0.20°, 17.91 ± 0.20°, 18.64 ± 0.20°, 20.57 ± 0.20°, 21.60 ± 0.20°, 22.37 ± 0.20°, 23.93 ± 0.20°, 24.54 ± 0.20° and 27.19 ± 0.20°.

[0181] In some embodiments of the application, the X-ray powder diffraction pattern of Form A is obtained using Cu Kα radiation and has characteristic diffraction peaks at 2θ angles of 9.01 ± 0.20°, 10.62 ± 0.20°, 13.27 ± 0.20°, 16.85 ± 0.20°, 17.91 ± 0.20°, 18.64 ± 0.20°, 20.57 ± 0.20°, 21.60 ± 0.20°, 22.37 ± 0.20°, 23.93 ± 0.20°, 24.54 ± 0.20°, and 27.19 ± 0.20°.

[0182] In some embodiments of the application, the X-ray powder diffraction pattern of Form A is obtained using Cu Kα radiation and has characteristic diffraction peaks at 2θ angles of 9.02 ± 0.20°, 13.29 ± 0.20°, 22.42 ± 0.20°, and / or 10.62 ± 0.20°, and / or 11.65 ± 0.20°, and / or 12.28 ± 0.20°, and / or 12.82 ± 0.20°, and / or 13.27 ± 0.20°, and / or 14.44 ± 0.20°, and / or 16.85 ± 0.20°, and / or 17.91 ± 0.20°, and / or 18.64 ± 0.20°, and / or 19.81 ± 0.20°, and / or 20.57 ± 0.20°, and / or 20.92 ± 0.20°, and / or 21.31 ± 0.20°, and / or 21.60 ± 0.20°, and / or 22.37 ± 0.20°, and / or 23.38 ± 0.20°, and / or 23.93 ± 0.20°, and / or 24.54 ± 0.20°, and / or 25.28 ± 0.20°, and / or 26.70 ± 0.20°, and / or 27.19 ± 0.20°, and / or 29.88 ± 0.20°, and / or 30.81 ± 0.20°.

[0183] In some embodiments of the application, the X-ray powder diffraction pattern of Form A is obtained using Cu Kα radiation and has characteristic diffraction peaks at 2θ angles of 9.01°, 10.62°, 12.28°, 12.82°, 13.27°, 16.85°, 17.91°, 18.08°, 18.64°, 19.81°, 20.57°, 20.92°, 21.31°, 21.60°, 22.37°, 23.38°, 23.93°, 24.54°, 25.28°, 27.19°, and 30.81°.

[0184] In some embodiments of the application, the X-ray powder diffraction pattern of the Form A is substantially as shown in Figure 3.

[0185] In some embodiments of the application, the X-ray powder diffraction pattern of the Form A is substantially as shown in Figure 3.

[0186] In some embodiments of the application, the X-ray powder diffraction pattern of the Form A is substantially as shown in Figure 3.

[0187] In some embodiments of the application, the X-ray powder diffraction pattern of the Form A is substantially as shown in Figure 3.

[0188] In some embodiments of the application, the differential scanning calorimetry curve (DSC) of the Form A has an endothermic peak with an onset value at 248.2±3°C.

[0189] In some embodiments of the application, the differential scanning calorimetry curve (DSC) of the Form A has an exothermic peak with an onset value at 181.3±3°C and an endothermic peak with an onset value at 248.2±3°C.

[0190] In some embodiments of the application, the DSC pattern of the Form A is substantially as shown in Figure 4.

[0191] In some embodiments of the application, the thermal gravimetric analysis curve (TGA) of the Form A has a weight loss of 0.75% in the range of 30.0±3°C to 220.0±3°C.

[0192] In some embodiments of the application, the thermal gravimetric analysis curve (TGA) of the Form A has a weight loss of 0.33% in the range of 30.0±3°C to 165.0±3°C and a further weight loss of 0.42% in the range of 165.0±3°C to 220.0±3°C.

[0193] In some embodiments of the application, the TGA pattern of the Form A is substantially as shown in Figure 5.

[0194] The application also provides the B crystal form of compound 3, wherein the B crystal form has a Cu Kα radiation X-ray powder diffraction pattern with characteristic diffraction peaks at the following 2θ angles: 8.45±0.20°, 9.98±0.20° and 13.60±0.20°.

[0195] In some technical solutions of the application, the B crystal form has a Cu Kα radiation X-ray powder diffraction pattern with characteristic diffraction peaks at the following 2θ angles: 8.45±0.20°, 9.98±0.20°, 13.60±0.20°, 19.22±0.20° and 23.91±0.20°.

[0196] In some solutions of the application, the B crystal form has a Cu Kα radiation X-ray powder diffraction pattern with characteristic diffraction peaks at the following 2θ angles: 8.45±0.20°, 9.98±0.20°, 13.60±0.20°, 18.13±0.20°, 19.22±0.20°, 21.94±0.20°, 23.91±0.20° and 24.77±0.20°.

[0197] In some solutions of the application, the B crystal form has a Cu Kα radiation X-ray powder diffraction pattern with characteristic diffraction peaks at the following 2θ angles: 8.45±0.20°, 9.98±0.20°, 13.60±0.20°, 18.13±0.20°, 19.22±0.20°, 21.94±0.20°, 23.91±0.20° and 24.77±0.20°.

[0198] In some solutions of the application, the B crystal form has a Cu Kα radiation X-ray powder diffraction pattern with characteristic diffraction peaks at the following 2θ angles: 8.45±0.20°, 9.98±0.20°, 13.60±0.20°, 18.13±0.20°, 19.22±0.20°, 21.94±0.20°, 23.91±0.20° and 24.77±0.20°.

[0199] In some embodiments of the application, the X-ray powder diffraction pattern of the B crystalline form described above has characteristic diffraction peaks at the following 2Θ angles: 8.45 ± 0.20°, 9.98 ± 0.20°, 11.88 ± 0.20°, 13.60 ± 0.20°, 18.13 ± 0.20°, 19.22 ± 0.20°, 21.94 ± 0.20°, 23.02 ± 0.20°, 23.91 ± 0.20°, 24.77 ± 0.20°, and 25.43 ± 0.20°.

[0200] In some embodiments of the application, the X-ray powder diffraction pattern of the B crystalline form described above has characteristic diffraction peaks at the following 2Θ angles: 8.45 ± 0.20°, 9.98 ± 0.20°, 13.60 ± 0.20°, and / or 11.88 ± 0.20°, and / or 16.91 ± 0.20°, and / or 18.13 ± 0.20°, and / or 19.22 ± 0.20°, and / or 20.02 ± 0.20°, and / or 21.26 ± 0.20°, and / or 21.76 ± 0.20°, and / or 21.94 ± 0.20°, and / or 22.15 ± 0.20°, and / or 23.02 ± 0.20°, and / or 23.91 ± 0.20°, and / or 24.25 ± 0.20°, and / or 24.77 ± 0.20°, and / or 25.43 ± 0.20°, and / or 26.01 ± 0.20°, and / or 28.89 ± 0.20°, and / or 29.61 ± 0.20°.

[0201] In some embodiments of the application, the X-ray powder diffraction pattern of the B crystalline form described above has characteristic diffraction peaks at the following 2Θ angles: 8.45 ± 0.20°, 9.98 ± 0.20°, 13.60 ± 0.20°, and / or 11.88 ± 0.20°, and / or 16.91 ± 0.20°, and / or 18.13 ± 0.20°, and / or 19.22 ± 0.20°, and / or 20.02 ± 0.20°, and / or 21.26 ± 0.20°, and / or 21.76 ± 0.20°, and / or 21.94 ± 0.20°, and / or 22.15 ± 0.20°, and / or 23.02 ± 0.20°, and / or 23.91 ± 0.20°, and / or 24.25 ± 0.20°, and / or 24.77 ± 0.20°, and / or 25.43 ± 0.20°, and / or 26.01 ± 0.20°, and / or 28.89 ± 0.20°, and / or 29.61 ± 0.20°.

[0202] In some embodiments of the application, the X-ray powder diffraction pattern of the B crystal form of compound 3, obtained using Cu Kα radiation, has characteristic peaks at the following 2θ angles: 8.45°, 9.98°, 10.61°, 11.88°, 13.60°, 14.67°, 15.04°, 15.61°, 16.36°, 16.91°, 17.73°, 18.13°, 19.22°, 20.02°, 20.26°, 21.26°, 21.76°, 21.94°, 22.15°, 23.02°, 23.91°, 24.25°, 24.77°, 25.43°, 26.01°, 26.38°, 26.91°, 27.36°, 27.64°, 28.21°, 28.89°, 29.20°, 29.61°, 30.22°, 30.50°, 30.68°, 31.84°, 32.61°, 33.55°, 34.10° and 34.67°.

[0203] In some embodiments of the application, the X-ray powder diffraction pattern of the B crystal form of compound 3, obtained using Cu Kα radiation, is substantially as shown in Figure 6.

[0204] In some embodiments of the application, the X-ray powder diffraction pattern of the B crystal form of compound 3, obtained using Cu Kα radiation, has characteristic peaks at the following 2θ angles: 8.45°, 9.98°, 10.61°, 11.88°, 13.60°, 14.67°, 15.04°, 15.61°, 16.36°, 16.91°, 17.73°, 18.13°, 19.22°, 20.02°, 20.26°, 21.26°, 21.76°, 21.94°, 22.15°, 23.02°, 23.91°, 24.25°, 24.77°, 25.43°, 26.01°, 26.38°, 26.91°, 27.36°, 27.64°, 28.21°, 28.89°, 29.20°, 29.61°, 30.22°, 30.50°, 30.68°, 31.84°, 32.61°, 33.55°, 34.10° and 34.67°.

[0205] Table 2: XRPD spectrum analysis data of the B crystal form of compound 3

[0206] In some embodiments of the application, the differential scanning calorimetry curve (DSC) of the B crystal form of compound 3 has an endothermic peak starting at 249.5±3°C.

[0207] In some embodiments of the application, the DSC spectrum of the B crystal form of compound 3 is substantially as shown in Figure 7.

[0208] In some embodiments of the application, the thermogravimetric analysis curve (TGA) of the B crystal form of compound 3 has a weight loss of 0.25% in the range of 30.0±3°C to 150.0±3°C.

[0209] In some embodiments of the application, the TGA spectrum of the B crystal form of compound 3 is substantially as shown in Figure 8.

[0210] The present application also provides a C crystal form of compound 3, wherein the X-ray powder diffraction pattern of the C crystal form of compound 3, obtained using Cu Kα radiation, has characteristic peaks at the following 2θ angles: 9.83±0.20°, 10.29±0.20°, 15.72±0.20°, 21.15±0.20° and 23.27±0.20°.

[0211] In some embodiments of the application, the X-ray powder diffraction pattern of the Form C, obtained using Cu Kα radiation, includes at least 6, 7, or 8 peaks selected from the group consisting of: 9.83 ± 0.20°, 10.29 ± 0.20°, 12.04 ± 0.20°, 15.72 ± 0.20°, 17.51 ± 0.20°, 21.15 ± 0.20°, 23.27 ± 0.20°, and 25.76 ± 0.20°, in terms of 2-theta.

[0212] In some embodiments of the application, the X-ray powder diffraction pattern of the Form C, obtained using Cu Kα radiation, has characteristic peaks at the following 2-theta angles: 9.83 ± 0.20°, 10.29 ± 0.20°, 12.04 ± 0.20°, 15.72 ± 0.20°, 17.51 ± 0.20°, 21.15 ± 0.20°, 23.27 ± 0.20°, and 25.76 ± 0.20°.

[0213] In some embodiments of the application, the X-ray powder diffraction pattern of the Form C, obtained using Cu Kα radiation, includes at least 10, 11, 12, or 13 peaks selected from the group consisting of: 9.83 ± 0.20°, 10.29 ± 0.20°, 11.65 ± 0.20°, 12.04 ± 0.20°, 15.72 ± 0.20°, 17.51 ± 0.20°, 19.55 ± 0.20°, 20.63 ± 0.20°, 21.15 ± 0.20°, 21.98 ± 0.20°, 23.27 ± 0.10°, 25.76 ± 0.20°, and 26.96 ± 0.10°, in terms of 2-theta.

[0214] In some embodiments of the application, the X-ray powder diffraction pattern of the Form C, obtained using Cu Kα radiation, has characteristic peaks at the following 2-theta angles: 8.16°, 9.83°, 10.29°, 11.27°, 11.65°, 12.04°, 12.62°, 13.36°, 13.91°, 14.46°, 14.77°, 15.72°, 16.76°, 17.51°, 18.02°, 19.55°, 20.09°, 20.63°, 21.15°, 21.98°, 22.29°, 22.73°, 23.27°, 23.43°, 24.47°, 24.71°, 25.41°, 25.76°, 26.96°, 28.00°, 29.53°, 29.97°, 30.26°, and 31.68°.

[0215] In some embodiments of the application, the X-ray powder diffraction pattern of the Form C, obtained using Cu Kα radiation, is substantially as shown in Figure 9.

[0216] In some embodiments of the present invention, the peak positions and relative intensities of the diffraction peaks in the X-ray powder diffraction patterns of Cu Kα radiation of the above-mentioned C-type crystal are shown in Table 3.

[0217] Table 3: XRPD spectrum analysis data of the C-crystal form of compound 3

[0218] In some technical solutions of the present invention, the differential scanning calorimetry (DSC) curve of the above-mentioned C crystal form has an initial value of endothermic peak at 250.9℃±3℃.

[0219] In some technical solutions of the present invention, the DSC spectrum of the above-mentioned C crystal form is basically as shown in Figure 10.

[0220] In some technical solutions of the present invention, the thermogravimetric analysis (TGA) curve of the above-mentioned C crystal form shows a weight loss of 2.60% in the range of 30.0±3℃ to 150.0±3℃.

[0221] In some technical solutions of the present invention, the TGA spectrum of the above-mentioned C crystal form is basically as shown in Figure 11.

[0222] The present invention also provides a hydrate of compound 3; preferably, the hydrate of compound 3 is shown in formula 3'.

[0223] The present invention also provides a D-type of compound 3', wherein the Cu Kα radiation X-ray powder diffraction pattern of the D-type has characteristic diffraction peaks at the following 2θ angles: 8.98±0.20°, 13.60±0.20°, 17.67±0.20°, 21.46±0.20° and 23.19±0.20°.

[0224] In some embodiments of the present invention, the X-ray powder diffraction pattern of Cu Kα radiation of the above-mentioned D crystal form, represented by the 2θ angle, contains at least 6, 7, or 8 diffraction peaks selected from the following: 8.98±0.20°, 13.60±0.20°, 17.23±0.20°, 17.67±0.20°, 18.48±0.20°, 21.46±0.20°, 21.87±0.20°, and 23.19±0.20°.

[0225] In some embodiments of the present invention, the X-ray powder diffraction pattern of Cu Kα radiation of the above-mentioned D crystal form has characteristic diffraction peaks at the following 2θ angles: 8.98±0.20°, 13.60±0.20°, 17.23±0.20°, 17.67±0.20°, 18.48±0.20°, 21.46±0.20°, 21.87±0.20° and 23.19±0.20°.

[0226] In some embodiments of the application, the X-ray powder diffraction pattern of the D crystal form, obtained using Cu Kα radiation, comprises at least 10, 11, 12, or 13 peaks selected from the group consisting of: 8.98 ± 0.20°, 13.60 ± 0.20°, 16.89 ± 0.10°, 17.23 ± 0.20°, 17.67 ± 0.20°, 18.48 ± 0.20°, 18.96 ± 0.20°, 21.46 ± 0.20°, 21.87 ± 0.20°, 23.19 ± 0.10°, 23.49 ± 0.10°, 23.98 ± 0.20°, and 26.88 ± 0.20° in terms of 2θ.

[0227] In some embodiments of the application, the X-ray powder diffraction pattern of the D crystal form, obtained using Cu Kα radiation, comprises characteristic peaks at the following 2θ values: 8.98 ± 0.20°, 13.60 ± 0.20°, 16.89 ± 0.10°, 17.23 ± 0.20°, 17.67 ± 0.20°, 18.48 ± 0.20°, 18.96 ± 0.20°, 21.46 ± 0.20°, 21.87 ± 0.20°, 23.19 ± 0.10°, 23.98 ± 0.20°, and 26.88 ± 0.20°.

[0228] In some embodiments of the application, the X-ray powder diffraction pattern of the D crystal form, obtained using Cu Kα radiation, comprises characteristic peaks at the following 2θ values: 8.98 ± 0.20°, 13.60 ± 0.20°, 16.89 ± 0.10°, 17.23 ± 0.20°, 17.67 ± 0.20°, 18.48 ± 0.20°, 18.96 ± 0.20°, 21.46 ± 0.20°, 21.87 ± 0.20°, 23.19 ± 0.10°, 23.98 ± 0.20°, and 26.88 ± 0.20°.

[0229] In some embodiments of the application, the X-ray powder diffraction pattern of the D crystal form, obtained using Cu Kα radiation, is substantially as shown in Figure 12.

[0230] In some embodiments of the application, the X-ray powder diffraction pattern of the D crystal form, obtained using Cu Kα radiation, comprises characteristic peaks at the following 2θ values: 8.98 ± 0.20°, 13.60 ± 0.20°, 16.89 ± 0.10°, 17.23 ± 0.20°, 17.67 ± 0.20°, 18.48 ± 0.20°, 18.96 ± 0.20°, 21.46 ± 0.20°, 21.87 ± 0.20°, 23.19 ± 0.10°, 23.98 ± 0.20°, and 26.88 ± 0.20°.

[0231] Table 4: XRPD spectrum analysis data of the D crystal form of compound 3

[0232] In some embodiments of the present application, the D crystal form described above has a differential scanning calorimetry curve (DSC) with an endothermic peak starting at 251.1°C±3°C.

[0233] In some embodiments of the present application, the D crystal form described above has a DSC spectrum substantially as shown in Figure 13.

[0234] In some embodiments of the present application, the D crystal form described above has a thermogravimetric analysis curve (TGA) with a weight loss of 5.16% in the range of 30.0°C±3°C to 120.0°C±3°C.

[0235] In some embodiments of the present application, the D crystal form described above has a TGA spectrum substantially as shown in Figure 14.

[0236] The crystalline compound 3 described in the present application can be in the form of a non-solvate or a solvate, such as a hydrate.

[0237] The present application provides a crystalline composition comprising a crystalline compound 3, wherein the crystalline compound 3 accounts for more than 50%, preferably more than 75%, more preferably more than 90%, and most preferably more than 95% by weight of the crystalline composition. The crystalline composition can also contain a small amount of other crystalline or non-crystalline forms of compound 3.

[0238] The present application provides a crystalline composition comprising a crystalline compound 3', wherein the crystalline compound 3' accounts for more than 50%, preferably more than 75%, more preferably more than 90%, and most preferably more than 95% by weight of the crystalline composition. The crystalline composition can also contain a small amount of other crystalline or non-crystalline forms of compound 3'.

[0239] The present application provides a pharmaceutical composition comprising a therapeutically effective amount of the compound described above, or a crystalline thereof, or a crystalline composition thereof; the pharmaceutical composition can comprise at least one pharmaceutically acceptable carrier or other excipient. In addition, the pharmaceutical composition of the present application can further comprise one or more other therapeutic agents.

[0240] The present application also provides the use of the compound described above, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, in the preparation of a medicament for treating a disease related to JAK1 / TYK2 inhibitors.

[0241] The present application also provides the use of the compound described above, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, or a crystalline thereof, in the preparation of a medicament for treating a disease related to JAK1 / TYK2 inhibitors.

[0242] The application also provides application of the above compound, stereoisomer thereof, or pharmaceutically acceptable salt thereof, or the B crystal form in preparation of a drug for treating a JAK1 / TYK2 inhibitor related disease.

[0243] In some technical solutions of the application, the JAK1 / TYK2 inhibitor related disease is an autoimmune disease, a neuroinflammatory disease, or a neurodegenerative disease.

[0244] In some technical solutions of the application, the autoimmune disease includes but is not limited to rheumatoid arthritis, vitiligo, Crohn's disease, ulcerative colitis, and the like.

[0245] In some technical solutions of the application, the neuroinflammatory disease and the neurodegenerative disease include but are not limited to Alzheimer's disease, Parkinson's syndrome, multiple sclerosis, amyotrophic lateral sclerosis, and the like.

[0246] In some technical solutions of the application, the neuroinflammatory disease and the neurodegenerative disease include but are not limited to Alzheimer's disease (and antibody treatment induced amyloid-related imaging abnormalities), Parkinson's syndrome, multiple sclerosis, and the like.

[0247] Technical effects

[0248] The compound of the application has strong inhibitory activity on JAK1 and TYK2 kinases, and the inhibitory activity on JAK2 and JAK3 is obviously weaker than that on JAK1, and has high selectivity; the compound has strong inhibitory activity on human peripheral blood mononuclear cell JAK1 and TYK2, and weak inhibitory activity on JAK2, and has high selectivity; the compound can significantly reduce the secretion of inflammatory factors (IL-6, TNF-alpha and CCL-2) in a cell inflammation model, and shows dose dependence; the free drug concentration ratio of the compound of the application in different species of plasma is high, and the compound has good drug properties; the compound has good stability in various species of liver microsomes and hepatocytes; the compound shows high permeability and low efflux in the MDCKII-MDR1 monolayer cell permeability experiment; the compound has no obvious inhibitory effect on the main five cytochrome P450 enzymes (CYP1A2, CYP2C9, CYP2C19, CYP2D6 and CYP3A4) of human liver microsomes, and has low risk of "drug-drug interaction"; the compound has no obvious induction effect on CAR, PXR and AhR, and has low induction risk of the main cytochrome P450 enzymes; the compound has no obvious inhibition on hERG; the compound shows high oral exposure and high oral bioavailability in various species of PK experiments, and has small species difference, and has excellent pharmacokinetic properties; the compound also has good brain tissue distribution, and has good brain entry; the compound shows significant therapeutic effect in an EAE efficacy model, has low onset dose, and shows dose-effect positive correlation, and exhibits excellent in vivo efficacy properties.

[0249] The crystal form of the compound of the present application has simple preparation process, stable physical and chemical properties, slight hygroscopicity, and good application prospect.

[0250] Definitions and Descriptions

[0251] The following terms and phrases, as used herein, are intended to have the following meanings, unless otherwise indicated. A particular term or phrase should not be construed as undefined or unclear, but should be understood according to its ordinary meaning. When a trade name appears herein, it is intended to refer to its corresponding product or active ingredient thereof.

[0252] The term "pharmaceutically acceptable" as used herein, with respect to compounds, materials, compositions, and / or dosage forms, means those that are, within the scope of sound medical judgment, suitable for use in contact with the tissues of humans and animals without excessive toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit / risk ratio.

[0253] The term "pharmaceutically acceptable salt" means a salt of a compound of the present application, which is discovered to have particular substituents, prepared from the compounds of the present application from relatively non-toxic, inorganic or organic acids or bases. When the compound of the present application contains relatively acidic functionalities, base addition salts can be obtained by contacting the neutral form of such compounds with a sufficient amount of the desired base to produce the salt. When the compound of the present application contains relatively basic functionalities, acid addition salts can be obtained by contacting the neutral form of such compounds with a sufficient amount of the desired acid to produce the salt. The pharmaceutically acceptable salts of the present application can be synthesized from the parent compound that contains an acidic or basic moiety by conventional chemical methods. Generally, the salt preparation methods are procedures by which a free acid or base form of these compounds is reacted with a stoichiometric amount of the appropriate base or acid in water or in an organic solvent or in both, to form the desired salt.

[0254] The compounds of the present application can exist in particular geometric or stereoisomeric forms. The present application contemplates all such compounds, including cis- and trans-forms, atropisomers, (-)- and (+)-enantiomers, (R)- and (S)-enantiomers, diastereomers, (D)- isomers, (L)-isomers, as well as racemic mixtures and other mixtures thereof, e.g., mixtures of enantiomers or diastereomers, all of which are intended to be within the scope of the present application. Additional asymmetric carbon atoms can be present in a substituent group. All such isomers, as well as mixtures thereof, are included within the scope of the present application. The optical purity of a single configuration compound can be expressed in terms of optical rotation, chiral purity, or ee, etc. Among them, chiral purity refers to the content determined by testing means (such as GC, HPLC, SFC, NMR, etc.); ee refers to the percentage of isomer excess or enantiomer excess, which is the difference between the percentage contents of two isomers or two enantiomers. For example, by SFC detection, the content of one isomer a is 90%, and the content of the other isomer b is 10%, then the chiral purity of isomer a is 90%, and the ee value is 80%.

[0255] The compounds of the present application can exist in particular tautomeric forms. Unless otherwise specified, the term "tautomer" or "tautomeric forms" refers to isomers that differ in the position or nature of a proton. Tautomers are interconvertible and are in dynamic equilibrium. For example, in solution, a chemical equilibrium of tautomers can be reached. For example, proton tautomers (also known as prototropic tautomers) include interconversions by proton migration, such as keto-enol isomerization and imine-enamine isomerization. Valence tautomers include interconversions by reorganization of some bonding electrons. For example, a specific example of keto-enol tautomerization is the interconversion between pentane-2,4-dione and 4-hydroxypent-3-en-2-one.

[0256] Unless otherwise specified, the term "enantiomer" or "optical isomer" refers to stereoisomers that are mirror images of one another.

[0257] Unless otherwise specified, the term "cis-trans isomer" or "geometric isomer" is caused by the double bond or ring-forming carbon atom single bond cannot rotate freely.

[0258] The term "diastereomer" means a stereoisomer of a molecule that has at least two chiral centers and exists as nonmirror-image relationships between molecules unless otherwise indicated. "(+)" indicates dextrorotation, "(-)" indicates levorotation, and "(±)" indicates racemic unless otherwise indicated.

[0259] Unless otherwise indicated, a wedge real line bond and a wedge dashed line bond indicate absolute stereochemistry about a stereocenter. A straight real line bond and a straight dashed line bond indicate relative stereochemistry about a stereocenter. A wavy line indicates a wedge real line bond and / or a wedge dashed line bond or a wavy line indicates a straight real line bond and / or a straight dashed line bond

[0260] Unless otherwise indicated, a carbon atom bearing an "*" is a chiral carbon atom, and exists in the form of a single enantiomer in the (R) or (S) form or enriched in one enantiomeric form. For example, indicates or or enriched in one enantiomeric form.

[0261] Unless otherwise indicated, the terms "enriched in one isomer," "isomer-enriched," "enriched in one enantiomer," or "enantiomer-enriched" mean that the content of one isomer or enantiomer is less than 100%, and the content of the isomer or enantiomer is greater than or equal to 60%, or greater than or equal to 70%, or greater than or equal to 80%, or greater than or equal to 90%, or greater than or equal to 95%, or greater than or equal to 96%, or greater than or equal to 97%, or greater than or equal to 98%, or greater than or equal to 99%, or greater than or equal to 99.5%, or greater than or equal to 99.6%, or greater than or equal to 99.7%, or greater than or equal to 99.8%, or greater than or equal to 99.9%.

[0262] Optically active (R)- and (S)-isomers and the D and L isomers can be prepared by chiral synthesis or chiral reagents or other conventional techniques. If one enantiomer of a compound of the application is desired, it can be prepared by asymmetric synthesis or derivatization with a chiral auxiliary, wherein the resulting diastereomeric mixture is separated and the auxiliary group cleaved to yield the pure desired enantiomer. Alternatively, when a basic functional group (such as an amino group) or an acidic functional group (such as a carboxyl group) is present in the molecule, a diastereomeric salt form of the compound with an appropriate optically active acid or base can be formed, and the desired enantiomer recovered by conventional means known in the art, such as elution from a column or fractional crystallization. Additionally, the separation of enantiomers and diastereomers is typically accomplished by the use of chromatography with a chiral stationary phase, optionally in combination with chemical derivatization (e.g., formation of a carbamate from an amine).

[0263] The compounds of the application can contain unnatural proportions of atomic isotopes at one or more of the atoms that constitute such compounds. For example, the compounds can be radiolabeled with radioactive isotopes, such as for example tritium ( 3 H), iodine-125 ( 125 I) or C-14 ( 14 C). For example, deuterium can be substituted for hydrogen to form deuterated drugs, which have advantages over non-deuterated drugs, such as reduced toxicity, increased stability, increased efficacy, increased biological half-life, etc. All isotopic variations of the compounds of the application, whether radioactive or not, are encompassed within the scope of the present application.

[0264] The term "substituted" means that any one or more hydrogen atoms on the particular atom is replaced with a substituent group, which can include variations of deuterium and hydrogen, provided that the valency of the particular atom is not exceeded and that the substituted compound is stable. When the substituent is oxygen (i.e., =0), it means that two hydrogen atoms are replaced.

[0265] The term "optionally" or "optionally" means that the subsequently described event or circumstance can or can not occur, and that the description includes instances where the event or circumstance occurs and instances where it does not.

[0266] The term "optionally substituted" means that the group can or can not be substituted and that the types and number of substituents are any that are chemically possible.

[0267] In some embodiments of the application, "optionally substituted with 1 or more R" means that the group can be unsubstituted or substituted, and the number of substituents R is 1 to 10, for example 1, 2, 3, 4, 5, 6, 7, or 8 R, or 1, 2, 3, 4, or 5 R, or 1, 2, or 3 R; when multiple R groups are present, each R can be the same or different.

[0268] When any variable (e.g., R) occurs more than one time in a compound or substituent, each occurrence of that variable is independent of the other. Thus, for example, if a group is substituted with 0-2 R, then that group can be optionally substituted with up to two R, and each occurrence of R is selected independently.

[0269] When the number of linking groups is 0, such as -(CRR)0-, it means that the linking group is a single bond.

[0270] When one of the variables is a single bond, it means that the two groups to which it is attached are directly connected, such as when L represents a single bond in A-L-Z, it means that the structure is actually A-Z.

[0271] When a substituent is null, it means that the substituent is not present, such as when X is null in A-X, it means that the structure is actually A.

[0272] When a recited substituent does not specify through which atom of the substituent it is bonded to the parent structure, the substituent can be bonded through any of its atoms, for example, a pyridyl group as a substituent can be bonded to the parent structure through any of the carbon atoms of the pyridyl ring. When a recited linking group does not specify its direction of attachment, its direction of attachment is arbitrary, for example, where the linking group L is -M-W-, then -M-W- can be attached to ring A and ring B to form either in the same direction as the reading order from left to right, or The combination of substituents and / or variables is permitted only if the combination results in a stable compound.

[0273] Unless otherwise specified, when a group has one or more points of attachment, any one or more of the points of the group can be attached to other groups by a chemical bond. When the manner of attachment of the chemical bond is not specified, and there is an H atom at the point of attachment, the number of H atoms at the point of attachment is reduced by one for each chemical bond that is formed. The chemical bond that attaches the point to other groups can be represented by a straight, solid line a straight, dashed line or a wavy line . When the straight, dashed line or wavy line is used to represent a point of attachment, it can be a single, double, or triple bond, etc. For example, the straight, solid line in -OCH3represents attachment to other groups through the oxygen atom in the group; the straight, dashed line in -NH2represents attachment to other groups through both ends of the nitrogen atom in the group; the wavy line in -Ph represents attachment to other groups through the 1 and 2 carbon atoms in the phenyl group; represents that any point of attachment on the piperidinyl group can be attached to other groups by one chemical bond, including at least the four ways of attachment, even though H atoms are drawn on -N-, but still include groups that are attached by one chemical bond, in which case one H is reduced by one to become a corresponding monovalent piperidinyl group.

[0274] Unless otherwise specified, C n-n+m or C n -C n+m includes any specific instance of n to n+m carbons, and also includes any range of n to n+m. For example, C 1-12 includes C1, C2, C3, C4, C5, C6, C7, C8, C9, C 10 , C 11 , and C 12 , etc., and also includes C 1- 3, C 1-6 , C 1-9 , C 3-6 , C 3-9 , C 3-12 , C 6-9 , C 6-12 , and C 9-12etc. Similarly, n to n+m membered ring means a ring having from n to n+m atoms on the ring, for example, 3-12 membered ring includes 3 membered, 4 membered, 5 membered, 6 membered, 7 membered, 8 membered, 9 membered, 10 membered, 11 membered, and 12 membered rings, etc., as well as 3-6 membered, 3-9 membered, 5-6 membered, 5-7 membered, 6-7 membered, 6-8 membered, and 6-10 membered rings, etc.

[0275] Unless otherwise specified, the term "halo" or "halogen" by itself or as part of another substituent is meant to include fluoro, chloro, bromo, or iodo.

[0276] Unless otherwise specified, the term "alkyl" by itself or in combination with other terms, means a straight-chain or branched-chain saturated carbon hydride group consisting of from 1 to 20 carbon atoms. It can be monovalent, divalent, or multivalent. The alkyl group includes C 1-10 alkyl, C 1-6 alkyl, C 1-5 alkyl, C 1-4 alkyl, C 1-3 alkyl, etc. Examples of alkyl groups include, but are not limited to, methyl (Me), methylene (CH2), methine (CH), ethyl (Et), propyl (including n-propyl and isopropyl), n-butyl, t-butyl, n-pentyl, etc. For example, in some embodiments of the application, the alkyl group is a C 1-6 alkyl, including C 1-2 , C 1-3 , C 1-4 , C 2-3 , C 2-4 , C 2-5 , C1, C2, C3, C4, C5, C6 alkyl, etc. In other embodiments of the application, the alkyl group is a C 1-4 alkyl, including C 1-2 , C 1-3 , C 2-3 , C 2-4 , C1, C2, C3, C4 alkyl, etc. In other embodiments of the application, the alkyl group is a C 1-3 alkyl, including C 1-2 , C 2-3 , C1, C2, C3 alkyl, etc.

[0277] Unless otherwise specified, the term "alkenyl" by itself or in combination with other terms, means a straight-chain or branched-chain carbon hydride group consisting of from 2 to 20 carbon atoms, containing at least one carbon-carbon double bond. It can be monovalent, divalent, or multivalent. The alkenyl group includes C 2-10 alkenyl, C 2-6 alkenyl, C 2-5 alkenyl, C 2-4 alkenyl, C2-3 Alkenyl groups, etc., examples of which include, but are not limited to, vinyl, propenyl, 1-butenyl, and cis-butadienyl. For example, in some technical solutions of the present invention, the alkenyl group is C... 2-6 alkenyl groups, which include C 2-3 C 2-4 C 2-5 C2, C3, C4, C5, C6 alkenyl groups, etc.; in other technical solutions of the present invention, the alkenyl group is C 2-4 alkenyl groups, which contain C 2-3 C2, C3, C4 alkenyl groups, etc.; in other technical solutions of the present invention, the alkenyl group is C 2-3 Alkenyl groups, including C2 and C3 alkenyl groups, etc.

[0278] Unless otherwise specified, the term "alkynyl" on its own or in combination with other terms refers to a straight-chain or branched hydrocarbon group consisting of 2 to 20 carbon atoms and containing at least one carbon-carbon triple bond. It can be monovalent, divalent, or polyvalent. The alkynyl group includes C... 2-10 alkynyl group, C 2-6 alkynyl group, C 2-5 alkynyl group, C 2-4 alkynyl group, C 2-3 Alkyne groups, etc., examples of which include, but are not limited to, ethynyl, propynyl, 1-butynyl, etc. For example, in some technical solutions of the present invention, the alkynyl group is C. 2-6 Alkyne groups, which include C 2-3 C 2-4 C 2-5 C2, C3, C4, C5, C6 alkynyl groups, etc.; in other technical solutions of the present invention, the alkynyl group is C 2-4 Alkyne group, which includes C 2-3 C2, C3, C4 alkynyl groups, etc.; in some other technical solutions of the present invention, the alkynyl group is C 2-3 Alkyne groups, including C2 and C3 alkynyl groups, etc.

[0279] Unless otherwise specified, the term "alkoxy" on its own, or in combination with other terms, refers to alkyl groups comprising 1 to 20 carbon atoms that are attached to the remainder of a molecule by an oxygen atom. It can be monovalent, divalent, or polyvalent. The alkoxy group includes C... 1-10 Alkoxy, C 1-6 Alkoxy, C 1-5 Alkoxy, C 1-4 Alkoxy, C 1-3 Alkoxy groups, etc., examples of which include, but are not limited to, methoxy, ethoxy, propoxy (including n-propoxy and isopropoxy), etc. For example, in some technical solutions of the present invention, the alkoxy group is C. 1-6 Alkoxy groups, which include C 1-2, C 1-3 , C 1-4 , C 2-3 , C 2-4 , C 2-5 , C1, C2, C3, C4, C5, C6alkoxy and the like; in other embodiments of the application, the alkoxy group is C 1-4 alkoxy including C 1-2 , C 1-3 , C 2-3 , C 2-4 , C1, C2, C3, C4alkoxy and the like; in other embodiments of the application, the alkoxy group is C 1-3 alkoxy including C 1-2 , C 2-3 , C1, C2, C3alkoxy and the like.

[0280] The term "alkylthio" by itself or in combination with other terms, means those alkyl groups containing from one to twenty carbon atoms attached to the rest of the molecule by a sulfur atom, unless otherwise specified. It can be monovalent, divalent, or multivalent. The alkylthio group includes C 1-10 alkylthio, C 1-6 alkylthio, C 1-5 alkylthio, C 1-4 alkylthio, C 1-3 alkylthio and the like, examples of alkylthio groups include, but are not limited to, methylthio, ethylthio, propylthio (including n- and isopropylthio), and the like. For example, in some embodiments of the application, the alkylthio group is C 1-6 alkylthio including C 1-2 , C 1-3 , C 1-4 , C 2-3 , C 2-4 , C 2-5 , C1, C2, C3, C4, C5, C6alkylthio and the like; in other embodiments of the application, the alkylthio group is C 1-4 alkylthio including C 1-2 , C 1-3 , C 2-3 , C 2-4 , C1, C2, C3, C4alkylthio and the like; in other embodiments of the application, the alkylthio group is C 1-3 alkylthio including C 1-2 , C 2-3 , C1, C2, C3alkylthio and the like.

[0281] Unless otherwise specified, the term "alkylamino" on its own or in combination with other terms refers to alkyl groups comprising 1 to 20 carbon atoms that are attached to the remainder of a molecule by a nitrogen atom. They can be monovalent, divalent, or polyvalent, including monoalkylamino and dialkylamino groups. The alkylamino group comprises C... 1-10 Alkylamino, C 1-6 Alkylamino, C 1-5 Alkylamino, C 1-4 Alkylamino, C 1-3 Alkylamino groups, etc., examples of which include, but are not limited to, -NHCH3, -N(CH3)2, -NHCH2CH3, -N(CH3)CH2CH3, -NHCH2CH2CH3, -NHCH(CH3)2, etc. For example, in some technical solutions of the present invention, the alkylamino group is C 1-6 Alkylamino, which includes C 1-2 C 1-3 C 1-4 C 2-3 C 2- 4. C 2-5 C1, C2, C3, C4, C5, C6 alkylamino groups, etc.; in some other technical solutions of the present invention, the alkylamino group is C1, C2, C3, C4, C5, C6, etc. 1-4 Alkylamino, which includes C 1-2 C 1-3 C 2-3 C 2-4 C1, C2, C3, C4 alkylamino groups, etc.; in some other technical solutions of the present invention, the alkylamino group is C 1-3 Alkylamino, which includes C 1-2 C 2-3 C1, C2, C3 alkylamino, etc.

[0282] Unless otherwise specified, the term "cycloalkyl" on its own or in combination with other terms refers to a saturated or partially unsaturated cyclic hydrocarbon group consisting of 3 to 20 carbon atoms. It can be monovalent, divalent, or polyvalent. The cycloalkyl group may optionally contain one or more carbon-carbon double or triple bonds, but all rings must not be aromatic. The cycloalkyl group can be a saturated cycloalkyl group (meaning all rings are saturated), or a cycloalkenyl group (meaning a monocyclic or polycyclic system containing at least one double bond), etc. The cycloalkyl group can be monocyclic or polycyclic (e.g., spirocyclic, fused, bridged rings), etc. The cycloalkyl group includes C... 3-10 cycloalkyl, C 3-8 cycloalkyl, C 3-7 cycloalkyl, C 3-6 cycloalkyl, C 3-5 cycloalkyl, C 4-6 cycloalkyl, C 5-8 cycloalkyl, C 6-8Cycloalkyl, examples of which include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cyclohexadienyl, and the like. For example, in some embodiments of the application, the cycloalkyl is C 3-6 Cycloalkyl, examples of which include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cyclohexadienyl, and the like. For example, in some embodiments of the application, the cycloalkyl is C 3-5 Cycloalkyl, examples of which include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cyclohexadienyl, and the like. For example, in some embodiments of the application, the cycloalkyl is C 4-5 Cycloalkyl, examples of which include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cyclohexadienyl, and the like. For example, in some embodiments of the application, the cycloalkyl is C 4-6 Cycloalkyl, examples of which include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cyclohexadienyl, and the like. For example, in some embodiments of the application, the cycloalkyl is C 3-8 Cycloalkyl, examples of which include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cyclohexadienyl, and the like. For example, in some embodiments of the application, the cycloalkyl is C 3-5 Cycloalkyl, examples of which include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cyclohexadienyl, and the like. For example, in some embodiments of the application, the cycloalkyl is C 4-5 Cycloalkyl, examples of which include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cyclohexadienyl, and the like. For example, in some embodiments of the application, the cycloalkyl is C 4-6 Cycloalkyl, examples of which include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cyclohexadienyl, and the like. For example, in some embodiments of the application, the cycloalkyl is C 3-10 Cycloalkenyl, which contains at least one carbon-carbon double bond, includes 3-5 membered, 4-5 membered, 4-6 membered, 5-6 membered, 5-7 membered, 5-8 membered, 5-9 membered, 5-10 membered cycloalkenyl, and the like.

[0283] Unless otherwise specified, the term "heterocycloalkyl," by itself or in combination with another term, means a saturated or partially unsaturated cyclic group consisting of 3 to 20 ring atoms, 1, 2, 3, 4, 5, 6, 7, or 8 of which are heteroatoms independently selected from the group consisting of O, S, and N, with the remainder being carbon atoms, wherein the carbon atoms are optionally oxidized (i.e., C(O)), the nitrogen atoms are optionally quaternized, and the nitrogen and sulfur heteroatoms are optionally oxidized (i.e., NO and S(O)2, p is 1 or 2), and wherein the heteroatoms can occupy any available position in the heterocycloalkyl group and the heterocycloalkyl group can be attached to the rest of the molecule at any available carbon atom. It can be monovalent, divalent, or multivalent. The heterocycloalkyl group can optionally contain one or more double or triple bonds, but none of the rings is aromatic. The heterocycloalkyl group can be a saturated heterocycloalkyl group (meaning all rings are saturated) or a heterocycloalkenyl group (meaning at least one carbon-carbon double bond is contained in a single ring or multiple ring system), and the like. The heterocycloalkyl group includes 3-10 membered heterocycloalkyl, 3-8 membered heterocycloalkyl, 3-7 membered heterocycloalkyl, 3-6 membered heterocycloalkyl, 3-5 membered heterocycloalkyl, 4-6 membered heterocycloalkyl, 5-8 membered heterocycloalkyl, 6-8 membered heterocycloalkyl, and the like. Examples of heterocycloalkyl groups include, but are not limited to, azetidinyl, oxetanyl, thietanyl, pyrrolidinyl, pyrazolidinyl, imidazolidinyl, tetrahydrothiophenyl (including tetrahydrothiophen-2-yl and tetrahydrothiophen-3-yl, and the like), tetrahydrofuranyl (including tetrahydrofuran-2-yl, and the like), tetrahydropyranyl, piperidinyl (including 1-piperidinyl, 2-piperidinyl, and 3-piperidinyl, and the like), piperazinyl (including 1-piperazinyl and 2-piperazinyl, and the like), morpholinyl (including 3-morpholinyl and 4-morpholinyl, and the like), dioxanyl, dithianyl, isoxazolidinyl, isothiazolidinyl, 1,2-oxazinyl, 1,2-thiazinyl, hexahydropyridazinyl, etc. For example, in some embodiments of the present application, the heterocycloalkyl is a 3-6 membered heterocycloalkyl, which includes 3-5 membered, 4-5 membered, 4-6 membered, 3 membered, 4 membered, 5 membered, 6 membered heterocycloalkyl, etc. The heterocycloalkyl is a 3-8 membered heterocycloalkyl, which includes 3-5 membered, 4-5 membered, 4-6 membered, 3 membered, 4 membered, 5 membered, 6 membered, 7 membered, 8 membered heterocycloalkyl, etc. In some embodiments of the present application, the heterocycloalkyl is a monocyclic 4-8 membered nitrogen-containing heterocycloalkyl, which means that it is a saturated or partially unsaturated monocyclic group with 4-8 ring atoms, at least one of the heteroatoms in the ring atoms is N atom, which includes 4-5 membered, 4-6 membered, 5-6 membered, 5-7 membered, 5-8 membered heterocycloalkyl, etc. In some embodiments of the present application, the heterocycloalkyl is a monocyclic 4-8 membered oxygen-containing heterocycloalkyl, which means that it is a saturated or partially unsaturated monocyclic group with 4-8 ring atoms, at least one of the heteroatoms in the ring atoms is O atom, which includes 4-5 membered, 4-6 membered, 5-6 membered, 5-7 membered, 5-8 membered heterocycloalkyl, etc.

[0284] Unless otherwise specified, the terms "heteroaromatic ring" and "heteroaryl" are used interchangeably, and the term "heteroaryl" by itself or in conjunction with other terms refers to a monocyclic group or a polycyclic ring system consisting of 5 to 20 ring atoms having a conjugated pi-electron system, 1, 2, 3, 4, 5, 6, 7, or 8 of which are heteroatoms independently selected from O, S, and N, with the remainder being carbon atoms. Where the nitrogen atom is optionally quaternized, and the nitrogen and sulfur heteroatoms can be optionally oxidized (i.e., NO and S(O)2, p is 1 or 2). The heteroaryl group can be attached to the remainder of the molecule through a heteroatom or carbon atom, and it can be monovalent, divalent, or multivalent. The heteroaryl group includes 5-6 membered, 5-8 membered, 5-9 membered, 5-10 membered, 6-8 membered, 6-9 membered, 6-10 membered, 5 membered, 6 membered, 7 membered, 8 membered, 9 membered, 10 membered heteroaryl. Examples of the heteroaryl group include, but are not limited to, pyrrolyl (including N-pyrrolyl, 2-pyrrolyl, and 3-pyrrolyl, etc.), pyrazolyl (including 2-pyrazolyl and 3-pyrazolyl, etc.), imidazolyl (including N-imidazolyl, 2-imidazolyl, 4-imidazolyl, and 5-imidazolyl, etc.), oxazolyl (including 2-oxazolyl, 4-oxazolyl, and 5-oxazolyl, etc.), triazolyl (1H-1, 2, 3-triazolyl, 2H-1, 2, 3-triazolyl, 1H-1, 2, 4-triazolyl, and 4H-1, 2, 4-triazolyl, etc.), tetrazolyl, isoxazolyl (3-isoxazolyl, 4-isoxazolyl, and 5-isoxazolyl, etc.), thiazolyl (including 2-thiazolyl, 4-thiazolyl, and 5-thiazolyl, etc.), furanyl (including 2-furanyl and 3-furanyl, etc.), thiophenyl (including 2-thiophenyl and 3-thiophenyl, etc.), pyridyl (including 2-pyridyl, 3-pyridyl, and 4-pyridyl, etc.), pyridazinyl, pyrazinyl, pyrimidinyl (including 2-pyrimidinyl and 4-pyrimidinyl, etc.), indolyl, indazolyl, pyrimidoimidazolyl, etc. For example, in some embodiments of the present application, the heteroaryl group is a 5-10 membered heteroaryl group, which includes 5-6 membered, 5-8 membered, 5-9 membered, 5 membered, 6 membered, 7 membered, 8 membered, 9 membered, 10 membered heteroaryl; in other embodiments of the present application, the heteroaryl group is a 5-6 membered heteroaryl group, which includes 5 membered and 6 membered heteroaryl.

[0285] Unless otherwise specified, the term "aromatic ring" is a cyclic group having a conjugated pi-electron system, which is overlaid by a delocalized pi-electron cloud between the atoms. In structural formulas, it can be written in the form of single and double bonds alternately, or it can be represented by a delocalized pi-electron cloud. For example, the structures represented by structural formulas are the same; the structures represented by structural formulas are the same; and The structures represented are identical. The aromatic ring can be a monocyclic or polycyclic system, wherein each ring in a polycyclic system is aromatic. Unless otherwise specified, the ring optionally contains 0, 1, or more heteroatoms or heterogroups independently selected from O, S, NH, and N.

[0286] The compounds of the present invention can be prepared by a variety of synthetic methods known to those skilled in the art, including the specific embodiments listed below, embodiments formed by combining them with other chemical synthetic methods, and equivalent substitutions known to those skilled in the art. Preferred embodiments include, but are not limited to, the embodiments of the present invention.

[0287] Unless otherwise stated, X-ray powder diffraction (XRPD) can detect changes in crystal form, crystallinity, and crystal structure, and is a commonly used method for identifying crystal forms. The peak positions of XRPD patterns depend primarily on the structure of the crystal form and are relatively insensitive to experimental details, while their relative peak heights depend on many factors related to sample preparation and instrument geometry. Therefore, in some embodiments, the crystal form of the present invention is characterized by an XRPD pattern with certain peak positions, which is essentially as shown in the XRPD patterns provided in the accompanying drawings. Furthermore, the measurement of 2θ in the XRPD pattern can be subject to experimental error; the measurement of 2θ in the XRPD pattern may vary slightly between different instruments and different samples, therefore the value of 2θ should not be considered absolute. Depending on the instrument used in this experiment, there is an error tolerance of ±0.20° for the diffraction peaks.

[0288] The chemical reactions in the specific embodiments of this invention are carried out in a suitable solvent, which must be suitable for the chemical changes of this invention and the reagents and materials required therefor. To obtain the compounds of this invention, it is sometimes necessary for those skilled in the art to modify or select the synthesis steps or reaction flow based on existing embodiments.

[0289] Unless otherwise stated, in the differential scanning calorimetry curves of the compounds of the present invention, upward indicates exothermic (Exo Up).

[0290] Unless otherwise stated, room temperature in this invention refers to 15°C to 35°C.

[0291] "Mammals" include humans and livestock such as laboratory mammals and domestic pets (e.g., cats, dogs, pigs, sheep, cattle, sheep, goats, horses, rabbits), as well as non-domesticated mammals such as wild mammals.

[0292] The term "pharmaceutical composition" refers to a formulation of the compound of this application with a medium generally accepted in the art for delivering a biologically active compound to a mammal, such as a human. The medium includes all pharmaceutically acceptable carriers for use therein. Pharmaceutical compositions facilitate the administration of compounds to a living organism.

[0293] The term "therapeutically effective amount" means an amount of a drug or a pharmaceutical agent that is sufficient to effect treatment for a condition, disease or disorder. The exact amount of an effective dose will depend on the particular active agent, the severity of the condition, the age of the subject and the general state of the patient's health. An appropriate effective dose can be determined by one of ordinary skill in the art using only routine experimentation.

[0294] The term "treatment" means administering a compound or formulation described herein to improve or eliminate a disease or one or more symptoms associated with the disease, and includes: 1) inhibiting the disease or condition, i.e., arresting its development; and 2) relieving the disease or condition, i.e., causing regression of the disease or condition.

[0295] In the present application, "pharmaceutically acceptable carrier" means a carrier that is not biologically or otherwise undesirable, i.e., the carrier can be administered to an organism without causing any undesirable biological effects or interacting in a deleterious manner with any of the active or inactive components of the composition.

[0296] The words "comprise" or "comprising" and variations thereof such as "comprises" or "comprising", will be understood to imply a non-exclusive inclusion. That is, a process, method, article or apparatus that comprises a list of elements is not necessarily limited to those elements, but can include other elements not expressly listed or inherent to such process, method, article, or apparatus.

[0297] The compounds of the present application can be identified by conventional means known to those skilled in the art. If the present application relates to the absolute configuration of a compound, the absolute configuration can be determined by conventional techniques. For example, single crystal X-ray diffraction (SXRD) can be used to determine the absolute configuration of a compound. A single crystal is grown and diffracted intensity data is collected using a Bruker D8 venture diffractometer with Cu Kα radiation. The crystal structure is then solved using direct methods (Shelxs97) to determine the absolute configuration.

[0298] Abbreviations used in the present application: prep-HPLC represents preparative high performance liquid chromatography; TLC represents thin layer chromatography; HPMC represents hydroxypropyl methylcellulose; Saline represents physiological saline; Solutol represents polyethylene glycol-15 hydroxystearate; tween 80 represents polysorbate 80; PEG 400 represents polyethylene glycol 400; HEPES represents 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid; HBSS represents Hank's balanced salt solution; NADPH represents reduced form of coenzyme II, also known as reduced nicotinamide adenine dinucleotide phosphate; PBS represents phosphate buffered saline.

[0299] The solvents used in the present application are commercially available. The compounds are named according to the conventional nomenclature rules in the art or using software, and the commercially available compounds are named according to the supplier's catalogue name. The solvents used in the present application are commercially available. The compounds are named according to the conventional nomenclature rules in the art or using software, and the commercially available compounds are named according to the supplier's catalogue name.​

[0300] Instruments and analytical methods of the present application

[0301] (1) X-ray powder diffractometer (XRPD) method

[0302] Take an appropriate amount of sample powder (about 20 mg) in a zero background sample disc, and detect XRPD. The instrument parameters are as follows:

[0303] Instrument model: BRUKER D8 ADVANCE

[0304] X-ray source: Cu Kα, Kα1: Kα2: Kα2 / Kα1 intensity ratio = 0.5

[0305] Light tube voltage / light tube current: 40 kV, 40 mA

[0306] Divergence slit: 0.6 mm

[0307] Scan range: 3°-45°

[0308] Scan mode: continuous scanning

[0309] Step size: 0.02°

[0310] Scan time per step: 0.1 s.

[0311] (2) Differential Scanning Calorimeter (DSC) and Thermal Gravimetric Analyzer (TGA)

[0312] The instrument parameters for DSC and TGA testing are shown in Table 5.

[0313] Table 5 DSC and TGA instrument parameters

[0314] (3) Dynamic Vapor Sorption (DVS) method

[0315] The DVS instrument parameters are shown in Table 6.

[0316] Table 6 DVS instrument parameters BRIEF DESCRIPTION OF DRAWINGS

[0317] Figure 1.1: Clinical score of animals in the EAE efficacy model (I)

[0318] Figure 1.2: Body weight change of animals in the EAE efficacy model

[0319] Figure 2: Clinical scores of animals in EAE efficacy model (II)

[0320] Figure 3: XRPD pattern of Form A of compound 3

[0321] Figure 4: DSC pattern of Form A of compound 3

[0322] Figure 5: TGA pattern of Form A of compound 3

[0323] Figure 6: XRPD pattern of Form B of compound 3

[0324] Figure 7: DSC pattern of Form B of compound 3

[0325] Figure 8: TGA pattern of Form B of compound 3

[0326] Figure 9: XRPD pattern of Form C of compound 3

[0327] Figure 10: DSC pattern of Form C of compound 3

[0328] Figure 11: TGA pattern of Form C of compound 3

[0329] Figure 12: XRPD pattern of Form D of compound 3'

[0330] Figure 13: DSC pattern of Form D of compound 3'

[0331] Figure 14: TGA pattern of Form D of compound 3'

[0332] Figure 15: DVS pattern of Form B of compound 3

[0333] Figure 16: DVS pattern of Form D of compound 3' DETAILED DESCRIPTION

[0334] The present application is described in detail below by way of Examples, but it is not meant to be limited by any of the Examples. The present application has been described in detail by specific embodiments, and specific examples thereof have also been disclosed, and it will be apparent to those skilled in the art that various changes and modifications can be made thereto without departing from the spirit and scope of the present application.

[0335] Example 1

[0336] Step 1: Compound 1-1 (1.2 g, 3.41 mmol) and compound 1-2 (574 mg, 4.09 mmol) were added into tert-butyl alcohol (12 mL) under nitrogen atmosphere, followed by the addition of N,N-diisopropylethylamine (1.54 g, 11.9 mmol). The reaction was heated to 100 °C for 1 h. After the reaction was completed, the reaction was cooled to room temperature, diluted with 40 mL of ethyl acetate, washed with water (40 mL x 3), saturated brine (50 mL) sequentially, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 10: 1-0: 1) to give compounds 1-3A and 1-3B.

[0337] Compound 1-3A was characterized by TLC with a developing agent of petroleum ether: ethyl acetate = 1: 1, Rf= 0.4; LCMS: 456.1 [M+H] f + 1 H NMR (400 MHz, CDC13) δ = 9.58 (br d, J = 7.6 Hz, 1H), 9.14 (s, 1H), 8.08 (d, J = 8.4 Hz, 2H), 7.62 (d, J = 4.0 Hz, 1H), 7.32 (d, J = 8.0 Hz, 2H), 6.64 (d, J = 4.0 Hz, 1H), 4.25-4.19 (m, 1H), 4.16-4.12 (m, 1H), 3.83 (dd, J = 1.6, 12.4 Hz, 1H), 3.80-3.72 (m, 1H), 2.71-2.49 (m, 2H), 2.41 (s, 3H), 2.25-2.12 (m, 1H), 1.98 (tt, J = 4.0, 13.2 Hz, 1H), 1.85-1.75 (m, 1H), 1.73-1.61 (m, 1H).

[0338] Compound 1-3B was characterized by TLC with a developing agent of petroleum ether: ethyl acetate = 1: 1, Rf= 0.3; LCMS: 456.1 [M+H] f + 1 ​​​​H NMR (400 MHz, CDC13) δ = 9.11 (s, 1H), 8.83 (br d, J = 7.8 Hz, 1H), 8.08 (d, J = 8.3 Hz, 2H), 7.66 (d, J = 4.2 Hz, 1H), 7.32 (d, J = 8.2 Hz, 2H), 6.82 (d, J = 4.2 Hz, 1H), 4.34-4.24 (m, 1H), 4.15-4.10 (m, 1H), 3.78-3.61 (m, 1H), 3.31 (t, J = 10.9 Hz, 1H), 2.71-2.56 (m, 2H), 2.41 (s, 3H), 2.05-1.98 (m, 1H), 1.80-1.63 (m, 2H).

[0339] Step 2: Compound 1-3A (0.7 g, 1.54 mmol) was added to methanol (7 mL) under nitrogen atmosphere, then 5% palladium on carbon (0.2 g) was added. After three times of hydrogen replacement, the mixture was stirred at 25 °C under hydrogen atmosphere (15 psi) for 1 h. After the reaction was completed, the reaction mixture was filtered, and the filtrate was concentrated under reduced pressure to give compound 1-4A. LCMS: 426.1 [M+H] + . 1 H NMR (400 MHz, CDC13) δ = 8.02 (d, J = 8.4 Hz, 2H), 7.86 (s, 1H), 7.52 (d, J = 4.0 Hz, 1H), 7.25 (d, J = 8.4 Hz, 2H), 6.64 (d, J = 4.0 Hz, 1H), 4.68 (br d, J = 8.8 Hz, 1H), 4.31-4.17 (m, 1H), 3.91 (br dd, J = 2.8, 4.0 Hz, 1H), 3.68-3.59 (m, 1H), 3.16 (t, J = 10.8 Hz, 1H), 2.60 (d, J = 6.0 Hz, 2H), 2.37 (s, 3H), 2.34-2.27 (m, 1H), 1.96 (br dd, J = 2.8, 13.2 Hz, 2H), 1.69-1.46 (m, 3H).

[0340] Step 3: Compound 1-4A (600 mg, 1.41 mmol) was added to acetic acid (12 mL) under nitrogen atmosphere, followed by the addition of tetramethylammonium carbonate (1.92 g, 14.1 mmol) and p-toluenesulfonic acid (24.28 mg, 0.14 mol) successively. The reaction was stirred at 25 °C for 8 h. After the reaction was completed, 80 mL of saturated sodium bicarbonate solution was added for quenching, and the mixture was extracted with ethyl acetate (80 mL x 3). The combined organic phase was washed with 180 mL of saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 1:1-1:0) to give compound 1-5A. LCMS: 466.1 [M+H] + . 1 H NMR (400 MHz, CDCl3) d = 8.65 (s, 1H), 8.09 (d, J = 8.4 Hz, 2H), 7.79 (d, J = 4.0 Hz, 1H), 7.25 (s, 2H), 6.79 (d, J = 4.0 Hz, 1H), 4.62 (dtd, J = 4.4, 8.0, 12.4 Hz, 1H), 4.22 (s, 3H), 4.06 (d, J = 8.4 Hz, 2H), 3.83 (dtd, J = 2.0, 5.6, 11.4 Hz, 1H), 2.66 (d, J = 6.0 Hz, 2H), 2.49 (dq, J = 4.4, 12.8 Hz, 1H), 2.36 (s, 3H), 2.21-2.06 (m, 2H), 1.84-1.68 (m, 1H).

[0341] Step 4: Potassium hydroxide (280 mg) was dissolved in methanol (5 mL) under nitrogen atmosphere, followed by the addition of compound 1-5A (200 mg, 430 pmol). The reaction was stirred at 25 °C for 4 h. After the reaction was completed, the pH was adjusted to neutral with 4 M hydrochloric acid solution, and the mixture was diluted with 60 mL of water, extracted with ethyl acetate (60 mL x 3), and the combined organic phase was washed with 120 mL of saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (dichloromethane:methanol = 10:0-10:1) to give compound 1A. LCMS: 312.1 [M+H] + . 1H NMR (400 MHz, DMSO-d6) δ = 11.73 (br s, 1H), 8.34 (s, 1H), 7.45 (t, J = 2.8 Hz, 1H), 6.71 (dd, J = 1.6, 3.2 Hz, 1H), 4.71 - 4.48 (m, 1H), 4.13 (s, 3H), 4.03 (d, J = 8.4 Hz, 2H), 3.94 - 3.81 (m, 1H), 2.94 - 2.84 (m, 1H), 2.82 - 2.73 (m, 1H), 2.48 - 2.35 (m, 1H), 2.16 - 1.89 (m, 2H), 1.76 - 1.60 (m, 1H). Compound 1A was analyzed by SFC (Column: Chiralcel OD-3 50*4.6mm I.D., 3pm; Mobile phase: A phase: supercritical CO2, B phase: isopropanol (0.05% diethylamine); Gradient (B%): 5%-40%) with retention time 1.700 min and chiral purity 96.74%.

[0342] Step 5: Refer to the above synthesis method, compound 1-3B was used as raw material to prepare compound 1B crude product, which was purified by prep-HPLC (Column: Waters Xbridge 150*25mm*5pm; Mobile phase: [water (NH3-H2O)-acetonitrile]; Gradient (acetonitrile%): 12%-42%) to compound 1B. LCMS: 312.1 [M+H] + ; 1 H NMR (400 MHz, DMSO-d6) δ = 11.62 (br s, 1H), 8.27 (s, 1H), 7.36 (t, J = 3.2 Hz, 1H), 6.61 (dd, J = 2.0, 3.6 Hz, 1H), 4.75 - 4.57 (m, 1H), 4.07 (s, 3H), 4.05 - 3.99 (m, 1H), 3.84 (dd, J = 4.8, 12.0 Hz, 1H), 3.42 - 3.33 (m, 1H), 3.02 - 2.92 (m, 1H), 2.88 - 2.77 (m, 1H), 2.37 - 2.24 (m, 1H), 2.05 - 1.82 (m, 2H), 1.81 - 1.68 (m, 1H). Compound 1B was analyzed by SFC (Column: Chiralcel OD-3 50*4.6mm I.D., 3pm; Mobile phase: A phase: supercritical CO2, B phase: isopropanol (0.05% diethylamine); Gradient (B%): 5%-40%) with retention time 1.773 min and chiral purity 99.08%.

[0343] Example 2

[0344] Step 1: Compound 2-2 (23.8 g, 140 mmol) was added into tetrahydrofuran (300 mL) under nitrogen atmosphere, and the mixture was cooled to -78 °C. n-Butyllithium (2.5 M, 51.3 mL, 128 mmol) was added dropwise. After the addition was completed, the reaction solution was reacted at -78 °C for 0.5 h. Compound 2-1 (30 g, 117 mmol) was dissolved in tetrahydrofuran (100 mL) and the solution was added dropwise to the reaction solution. The mixture was stirred at -78 °C for 3 h. The reaction was quenched by adding 100 mL of saturated aqueous ammonium chloride solution at 0 °C, and extracted with ethyl acetate (100 mL x 2). The combined organic phase was washed with 100 mL of saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to give compound 2-3. 1 H NMR (400 MHz, CDCl3) δ = 5.09 (br d, J = 7.2 Hz, 1H), 4.46 (s, 2H), 4.32-4.25 (m, 1H), 4.20 (q, J = 7.2 Hz, 2H), 2.77-2.61 (m, 2H), 2.25-2.13 (m, 1H), 2.02-1.91 (m, 1H), 1.44 (s, 9H), 1.32-1.25 (m, 3H), 0.91 (s, 9H), 0.13 (s, 6H).

[0345] Step 2: Compound 2-3 (49 g, 114 mmol) was added into DMF (400 mL) under nitrogen atmosphere at 0 °C, and hydrazine hydrate (8.7 g, 165 mmol) was added dropwise. The reaction solution was warmed to 25 °C and stirred for 2 h. After the reaction was completed, the reaction solution was diluted with 100 mL of water, extracted with ethyl acetate (100 mL x 2), and the combined organic phase was washed with 100 mL of saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to give compound 2-4. 1 H NMR (400 MHz, CDCl3) δ = 6.01 (s, 1H), 5.28 (br d, J = 8.4 Hz, 1H), 4.74 (s, 2H), 4.40-4.28 (m, 1H), 4.18 (q, J = 7.2 Hz, 2H), 2.81-2.65 (m, 2H), 2.19-2.07 (m, 1H), 1.91 (br dd, J = 7.2, 12.8 Hz, 1H), 1.46 (s, 9H), 1.26 (t, J = 7.1 Hz, 3H), 0.92 (s, 9H), 0.10 (s, 6H).

[0346] Step 3: Compound 2-4 (50 g, 113 mmol) was added to a mixture of tetrahydrofuran (400 mL) and methanol (60 mL) under nitrogen atmosphere. Sodium borohydride (8.99 g, 238 mmol) was added portionwise at 0 °C. The reaction was allowed to warm to 25 °C and stirred for 12 h. The reaction was quenched by the addition of 50 mL of saturated aqueous ammonium chloride solution. The reaction was extracted with ethyl acetate (100 mL x 2). The combined organic phase was washed with 100 mL of saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to give the crude product. The crude product was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 10 / 1 to 0 / 1) to give compound 2-5. 1 H NMR (400 MHz, CDC13) δ = 5.99 (s, 1H), 5.02 (br d, J = 6.8 Hz, 1H), 4.73 (s, 2H), 3.73-3.62 (m, 2H), 3.61-3.55 (m, 1H), 2.84-2.72 (m, 1H), 2.70-2.54 (m, 1H), 1.84 (q, J = 6.8 Hz, 2H), 1.46 (s, 9H), 0.92 (s, 9H), 0.10 (s, 6H).

[0347] Step 4: Compound 2-5 (22 g, 55.1 mmol) and 1,1-azobis(cyclohexanecarbonitrile) (23.3 g, 92.5 mmol) were added to tetrahydrofuran (200 mL) under nitrogen atmosphere at 0 °C. Then tri-tert-butylphosphine (18.7 g, 92.5 mmol) was added dropwise slowly. After the addition was completed, the reaction was allowed to warm to 25 °C and stirred for 12 h. The reaction was quenched by the addition of 100 mL of saturated aqueous ammonium chloride solution. The reaction was extracted with ethyl acetate (100 mL x 2). The combined organic phase was washed with 100 mL of saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to give the crude product. The crude product was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 20 / 1 to 1 / 1) to give compound 2-6. 1 H NMR (400 MHz, CDC13) δ = 6.02 (s, 1H), 4.72 (br d, J = 2.8 Hz, 1H), 4.67 (s, 2H), 4.36-4.18 (m, 2H), 3.98-3.82 (m, 1H), 2.86 (br t, J = 6.4 Hz, 2H), 2.04-1.99 (m, 1H), 1.97-1.83 (m, 1H), 1.45 (s, 9H), 0.92 (s, 9H), 0.10 (d, J = 0.7 Hz, 6H).

[0348] Step 5: To a solution of compound 2-6 (5 g, 13.1 mmol) in tetrahydrofuran (50 mL) was added tetrabutylammonium fluoride (1 M in tetrahydrofuran, 14.4 mL, 14.4 mmol) at 0 °C under nitrogen atmosphere. The reaction was allowed to warm to 25 °C and stirred for 1 h. After completion of the reaction, the reaction was diluted with 50 mL of water and extracted with ethyl acetate (100 mL x 2). The combined organic phase was washed with 100 mL of saturated brine, dried over anhydrous sodium sulfate, filtered and the filtrate was concentrated under reduced pressure to obtain compound 2-7.

[0349] Step 6: Manganese dioxide (1.38 g, 131 mmol) was added to a solution of compound 2-7 (3.5 g, 13.1 mmol) in dichloromethane (25 mL) and methanol (25 mL) at room temperature under nitrogen atmosphere. The mixture was heated to 65 °C and stirred for 12 h. The reaction was allowed to cool to room temperature, filtered and the filtrate was concentrated under reduced pressure to obtain the crude product which was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 20 / 1 to 1 / 1) to obtain compound 2-8. The retention time of compound 2-8 was 0.939 min and the chiral purity was 98.87% by SFC (Chromatographic column: Chiralpak AD-3 50 x 4.6 mm I.D., 3 pm, Mobile phase: A phase: supercritical CO2, B phase: ethanol (0.05% diethylamine); Gradient (B%): 10% - 60%). 1 H NMR (400 MHz, CDC13) d = 9.92 (s, 1H), 6.55 (s, 1H), 4.71 (br s, 1H), 4.47 (dd, J = 4.8, 13.2 Hz, 1H), 4.35 - 4.24 (m, 1H), 4.06 (br dd, J = 6.8, 12.8 Hz, 1H), 2.94 (br t, J = 6.4 Hz, 2H), 2.12 (dtd, J = 2.8, 6.8, 13.2 Hz, 1H), 1.96 (qd, J = 6.6, 13.6 Hz, 1H), 1.47 (s, 9H).

[0350] Step 7: Sodium acetate (232 mg, 2.83 mmol) and hydroxylamine hydrochloride (157 mg, 2.26 mmol) were added to a solution of compound 2-8 (500 mg, 1.88 mmol) in methanol (5 mL) under nitrogen atmosphere. The reaction was stirred at 25 °C for 0.5 h. After completion of the reaction, the reaction was diluted with 10 mL of water and extracted with ethyl acetate (100 mL x 2). The combined organic phase was washed with 100 mL of saturated brine, dried over anhydrous sodium sulfate, filtered and the filtrate was concentrated under reduced pressure to obtain compound 2-9. LCMS: 281.1 [M+1] + .

[0351] Step 8: Under nitrogen atmosphere, boroges reagent (663 mg, 2.78 mmol) was slowly added to a solution of compound 2-9 (520 mg, 1.86 mmol) in dichloromethane (5 mL) at 0 °C, the reaction was warmed to 25 °C and stirred for 12 h. After the reaction was completed, 5 mL of water was added to quench the reaction, extracted with ethyl acetate (100 mL x 2), the combined organic phase was washed with 100 mL of saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to give compound 2-10. 1 H NMR (400 MHz, CDC13) δ = 6.42 (s, 1H), 4.69 (br d, J = 5.6 Hz, 1H), 4.43 (dd, J = 4.8, 13.2 Hz, 1H), 4.26 (br s, 1H), 4.02 (dd, J = 6.8 13.2 Hz, 1H), 3.00-2.84 (m, 2H), 2.11 (dtd, J = 2.8, 6.8, 13.2 Hz, 1H), 1.96 (td, J = 7.2, 14.0 Hz, 1H), 1.46 (s, 9H).

[0352] Step 9: Under nitrogen atmosphere, trimethylsilyl iodide (417 mg, 2.08 mmol) was slowly added to a solution of compound 2-10 (420 mg, 1.60 mmol) in dichloromethane (5 mL) at 0 °C, the reaction was stirred at 0 °C for 0.5 h. After the reaction was completed, the reaction was concentrated under reduced pressure at room temperature to give the crude of compound 2-11 hydroiodide salt.

[0353] Step 10: Under nitrogen atmosphere, compound 2-11 (438 mg, crude hydroiodide salt) and compound 2-12 (300 mg, 1.51 mmol) were added to a solution of isopropanol (8 mL), then N, N-diisopropylethylamine (1.37 g, 10.6 mmol) was added. The reaction was warmed to 80 °C and stirred for 12 h. After the reaction was completed, the reaction was cooled to room temperature, diluted with 5 mL of water, extracted with ethyl acetate (100 mL x 2), the combined organic phase was washed with 100 mL of saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to give the crude, which was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 10 / 1 to 0 / 1) to give compound 2-13. The retention time of compound 2-13 was 2.087 min and the chiral purity was 100% by SFC (Chromatographic column: Chiralcel OJ-3 50 x 4.6 mm I.D., 3 pm, mobile phase: A phase: supercritical CO2, B phase: ethanol (0.05% diethylamine); gradient (B%): 10%-60%). 1H NMR (400 MHz, CDC13) δ = 9.29 (s, 1H), 8.78 (br d, J = 8.0 Hz, 1H), 7.96 (d, J = 2.0 Hz, 1H), 7.04 (d, J = 2.0 Hz, 1H), 6.53 (s, 1H), 5.23-5.09 (m, 1H), 4.71 (dd, J = 4.8, 13.2 Hz, 1H), 4.28 (dd, J = 7.2, 13.2 Hz, 1H), 3.21-3.01 (m, 2H), 2.49-2.37 (m, 1H), 2.28-2.17 (m, 1H).

[0354] Step 11: Palladium on carbon (295 mg, 5%) was added to a mixture of compound 2-13 (300 mg, 925 μmol) in dichloromethane (4 mL) and methanol (4 mL) under nitrogen atmosphere. After three times of hydrogen replacement, the reaction was stirred at 25 °C under hydrogen atmosphere (15 psi) for 0.5 h. The reaction was filtered and the filtrate was concentrated under reduced pressure to give compound 2-14. LCMS: 295.1 [M+1] + .

[0355] Step 12: Compound 2-14 (100 mg, 340 μmol) was added to a mixture of toluene (0.5 mL) and tetrahydrofuran (0.5 mL) under nitrogen atmosphere at room temperature, followed by the addition of trimethyl orthoacetate (81.7 mg, 680 μmol) and p-toluenesulfonic acid (5.85 mg, 34.0 μmol) successively. The reaction was stirred at 100 °C for 1 h. The reaction was cooled to room temperature, diluted with 5 mL of water, extracted with ethyl acetate (50 mL x 2), and the combined organic phase was washed with 50 mL of saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to give a crude product, which was purified by silica gel column chromatography (dichloromethane / methanol = 1 / 0-10 / 1) to give compound 2. LCMS: 319.1 [M+1] + ; 1H NMR (400 MHz, DMSO-d6) δ = 8.83 (s, 1H), 8.24 (d, J = 2.2 Hz, 1H), 7.22 (d, J = 2.2 Hz, 1H), 6.88 (s, 1H), 5.42-5.22 (m, 1H), 4.85 (dd, J = 6.0, 12.4 Hz, 1H), 4.68-4.50 (m, 1H), 3.26-3.04 (m, 2H), 2.74 (s, 3H), 2.72-2.64 (m, 1H), 2.38-2.29 (m, 1H). SFC detection (Chiralcel OD-3 50*4.6 mm I.D., 3 pm, mobile phase: A phase is supercritical CO2, B phase is isopropanol / acetonitrile (0.05% diethylamine); gradient (B%): 20%-60%) the retention time of compound 2 was 1.073 min, the chiral purity was 99.89%; Example 2

[0356] Step 13: Refer to the above steps, compound 2-1A was used as raw material to prepare compound 2A. LCMS: 319.1 [M+1] + ; 1 H NMR (400 MHz, DMSO-d6) δ = 8.83 (s, 1H), 8.24 (d, J = 2.2 Hz, 1H), 7.22 (d, J = 2.2 Hz, 1H), 6.88 (s, 1H), 5.42-5.22 (m, 1H), 4.85 (dd, J = 6.0, 12.4 Hz, 1H), 4.68-4.50 (m, 1H), 3.26-3.04 (m, 2H), 2.74 (s, 3H), 2.72-2.64 (m, 1H), 2.38-2.29 (m, 1H). SFC detection (Chiralcel AD-3 50*4.6 mm I.D., 3 pm, mobile phase: A phase is supercritical CO2, B phase is ethanol (0.05% diethylamine); gradient (B%): 10%-60%) the retention time of compound 2A was 2.150 min, the chiral purity was 100%; the retention time of compound 2 was 1.418 min using this method.

[0357] Example 3

[0358] Step 1: Compound 3-2 (11.2 g, 46.4 mmol) was added to a solution of compound 1 (10 g, 46.4 mmol) in N,N-dimethylacetamide (100 mL) under nitrogen atmosphere, and the reaction was heated to 70 °C for 13 h. The reaction was cooled to room temperature, and the reaction was added to ice water (150 mL) and extracted with ethyl acetate (100 mL x 3). The combined organic phase was washed with water (50 mL x 3), saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to give a crude product, which was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 1:0-3:1) to give compound 3-3. LCMS: 183.1 [M+H-56] + .

[0359] Step 2: To a solution of compound 3-3 (10 g, 42.0 mmol) in methanol (100 mL) was added palladium on carbon (2 g, 5% content) under nitrogen atmosphere, and the reaction was stirred at 25 °C for 15 h after hydrogen was replaced for three times under hydrogen atmosphere (15 psi). The reaction was filtered, and the filtrate was concentrated under reduced pressure. To the residue was added a mixture of ethyl acetate / petroleum ether (V / V = 5 / 1, 30 mL), and the mixture was stirred at room temperature for 30 min, filtered, and the filter cake was dried under vacuum to give compound 3-4. 1 H NMR (400 MHz, CDCl3) d = 4.30 (s, 1H), 4.16-4.02 (m, 1H), 3.74-3.40 (m, 2H), 3.04 (t, J = 10.8 Hz, 1H), 2.68-2.43 (m, 2H), 2.24-2.06 (m, 1H), 1.93-1.80 (m, 1H), 1.64-1.39 (m, 10H), 1.39-1.20 (m, 1H); two-dimensional NMR spectrum confirmed the structure of compound 3-4.

[0360] Step 3: Trimethylsilyl iodide (3.30 g, 16.5 mmol) was added to a solution of compound 3-4 (3.6 g, 15.0 mmol) in dichloromethane (40 mL) at 0 °C under nitrogen atmosphere, and the reaction was stirred at 0 °C for 1 h. The reaction was concentrated under reduced pressure to give compound 3-5 as a crude product.

[0361] Step 4: To a solution of compound 1-1 (3.82 g, 10.8 mmol) and compound 3-5 (1.52 g, 10.8 mmol) in isopropanol (40 mL) was added N,N-diisopropylethylamine (4.20 g, 32.5 mmol) under nitrogen atmosphere. The reaction was heated to 90 °C for 2 hours. The reaction was cooled to room temperature and concentrated. The residue was dissolved in ethyl acetate (100 mL) and washed with water (20 mL), saturated brine (20 mL) successively. The organic layer was dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give a crude product. The crude product was purified by column chromatography on silica gel (petroleum ether: ethyl acetate = 10: 1-1: 1) to give compound 3-6. LCMS: 456.0 [M+H] + . 1 H NMR (400 MHz, CDC13) d = 9.11 (s, 1H), 8.83 (d, J = 8.0 Hz, 1H), 8.07 (d, J = 8.4 Hz, 2H), 7.66 (d, J = 4.0 Hz, 1H), 7.32 (d, J = 8.0 Hz, 2H), 6.82 (d, J = 4.4 Hz, 1H), 4.36-4.25 (m, 1H), 4.16-4.06 (m, 1H), 3.79-3.64 (m, 1H), 3.31 (t, J = 11.0 Hz, 1H), 2.71-2.55 (m, 2H), 2.49-2.34 (m, 4H), 2.05-1.96 (m, 1H), 1.81-1.63 (m, 2H).

[0362] Step 5: To a solution of compound 3-6 (4.65 g, 10.2 mmol) in methanol (50 mL) was added palladium on carbon (1 g, 5% loading) under nitrogen atmosphere. After three times of hydrogen replacement, the reaction was stirred under hydrogen atmosphere (15 psi) at 25 °C for 15 hours. The reaction was filtered and the filtrate was concentrated under reduced pressure to give compound 3-7. LCMS: 426.0 [M+H] + ; 1H NMR (400 MHz, CDC13) δ = 8.02 (d, J = 8.0 Hz, 2H), 7.85 (s, 1H), 7.51 (d, J = 4.0 Hz, 1H), 7.25 (d, J = 8.0 Hz, 2H), 6.63 (d, J = 4.0 Hz, 1H), 4.65 (d, J = 8.4 Hz, 1H), 4.35 - 4.13 (m, 1H), 3.97 - 3.78 (m, 1H), 3.70 - 3.56 (m, 1H), 3.16 (t, J = 10.8 Hz, 1H), 2.59 (d, J = 5.6 Hz, 2H), 2.51 - 2.40 (m, 1H), 2.36 (s, 3H), 2.34 - 2.16 (m, 2H), 2.00 - 1.90 (m, 1H), 1.72 - 1.58 (m, 1H), 1.57 - 1.42 (m, 1H).

[0363] Step 6: To a solution of compound 3-7 (800 mg, 1.88 mmol) in toluene / tetrahydrofuran (V / V = 1 / 1, 16 mL) was added trimethyl orthoacetate (451.79 mg, 3.76 mmol) and p-toluenesulfonic acid (32.38 mg, 188.01 μmol) under nitrogen atmosphere. The reaction was stirred at 100 °C for 1 h. The reaction was cooled to room temperature, diluted with water (50 mL), extracted with ethyl acetate (50 mL x 3), the combined organic phase was washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, filtered and the filtrate was concentrated under reduced pressure to give a crude product, which was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 1:1-0:1) to give compound 3-8. LCMS: 450.1 [M+H] + ; 1 H NMR (400 MHz, Chloroform-d) δ = 8.81 (s, 1H), 8.11 (d, J = 8.4 Hz, 2H), 7.84 (d, J = 4.0 Hz, 1H), 7.28 (s, 1H), 7.26 (s, 1H), 6.88 (d, J = 4.0 Hz, 1H), 4.62 (br s, 1H), 4.20 - 4.14 (m, 1H), 4.09 - 4.02 (m, 1H), 4.01 - 3.91 (m, 1H), 2.72 - 2.71 (m, 3H), 2.70 - 2.65 (m, 1H), 2.63 - 2.50 (m, 1H), 2.36 (s, 3H), 2.24 - 2.13 (m, 2H), 1.93 - 1.78 (m, 2H).

[0364] Step 7: To a solution of compound 3-8 (800 mg, 1.78 mmol) in methanol (8 mL) was added potassium hydroxide aqueous solution (2 M, 8 mL) under nitrogen atmosphere, the reaction was heated to 65 °C and stirred for 2 h. The reaction was cooled to room temperature, diluted with water (50 mL), extracted with ethyl acetate (50 mL x 3), the combined organic phase was washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, filtered and the filtrate was concentrated under reduced pressure to give a crude product, which was purified by column chromatography on silica gel (dichloromethane:methanol = 1:0-10:1) to give compound 3. LCMS: 296.1 [M+H] + ; 1 H NMR (400 MHz, DMSO-d6) d = 11.85 (br s, 1H), 8.47 (s, 1H), 7.47 (t, J = 3.2 Hz, 1H), 6.81 (br s, 1H), 4.76-4.48 (m, 1H), 4.20-4.12 (m, 1H), 4.11-3.99 (m, 2H), 2.98-2.87 (m, 1H), 2.84-2.75 (m, 1H), 2.66 (s, 3H), 2.62-2.52 (m, 1H), 2.17-2.07 (m, 1H), 2.02 (br d, J = 13.1 Hz, 1H), 1.83-1.63 (m, 1H). Analyzed by SFC (analysis method: column: Chiralcel OJ-3 50*4.6mm I.D., 3 pm; mobile phase: A phase: supercritical CO2, B phase: isopropanol (0.05% diethylamine); gradient (B%): 5%-40%), the retention time of compound 3 was 1.513 min, and the chiral purity was 100%.

[0365] Example 4

[0366] Step 1-2: Compound 4-3 was prepared from compound 4-1 and compound 1-2 according to the method of Example 1.

[0367] Step 3: To a solution of compound 4-3 (300 mg, 747.06 pmol) in toluene (10 mL) was added N,N-thiocarbonyldiimidazole (266.27 mg, 1.49 mmol) under nitrogen atmosphere, the reaction was heated to 80 °C and stirred for 1 h. The reaction was cooled to room temperature, diluted with ethyl acetate (120 mL), washed with water (100 mL x 3) and saturated brine (120 mL) successively, dried over anhydrous sodium sulfate, filtered and the filtrate was concentrated under reduced pressure to give a crude product, which was purified by column chromatography on silica gel (petroleum ether: ethyl acetate = 2:1-1:2) to give compound 4-4. LCMS: 444.1 [M+H] + ;1 H NMR (400 MHz, CDC13) δ = 10.54 (br s, 1H), 8.35 (br s, 1H), 7.52 (d, J = 3.6 Hz, 1H), 6.82 (d, J = 3.6 Hz, 1H), 5.76 (s, 2H), 4.48-3.94 (m, 3H), 3.64-3.42 (m, 2H), 2.68 (br d, J = 5.6 Hz, 2H), 2.18 (br d, J = 12.0 Hz, 2H), 1.97-1.67 (m, 3H), 0.99-0.85 (m, 2H), -0.05 (s, 9H).

[0368] Step 4: To a solution of compound 4-4 (300.00 mg, 676.23 pmol) and potassium carbonate (140.19 mg, 1.01 mmol) in N,N-dimethylformamide (5 mL) was added iodomethane (143.98 mg, 1.01 mmol) under nitrogen atmosphere. The reaction was stirred at 25 °C for 1 h. To the reaction was added ethyl acetate (80 mL), which was washed with water (70 mL x 3) and saturated brine (100 mL) successively. The organic phase was dried over anhydrous sodium sulfate, filtered and the filtrate was concentrated under reduced pressure to give a crude product, which was purified by column chromatography on silica gel (petroleum ether: ethyl acetate = 1: 1-1:2) to give compound 4-5. LCMS: 458.1 [M+H] + .

[0369] Step 5: To a solution of compound 4-5 (250 mg, 546.25 pmol) in dichloromethane (5 mL) was added m-chloroperoxybenzoic acid (267 mg, 1.32 mmol, 85% purity) portionwise. After the addition, the reaction was stirred at 25 °C for 1 h. To the reaction was added 80 mL of dichloromethane, which was washed with water (70 mL x 3) and saturated brine (100 mL) successively. The organic phase was dried over anhydrous sodium sulfate, filtered and the filtrate was concentrated under reduced pressure to give a crude product, which was purified by column chromatography on silica gel (petroleum ether: ethyl acetate = 2: 1-1:2) to give compound 4-6. LCMS: 490.2 [M+H] + ; 1H NMR (400 MHz, CDC13) δ = 8.87 (s, 1H), 7.53 (d, J = 3.6 Hz, 1H), 6.88 (d, J = 3.6 Hz, 1H), 5.82 (s, 2H), 5.50-5.27 (m, 1H), 4.48-4.34 (m, 1H), 4.24 (dd, J = 4.4, 10.4 Hz, 1H), 4.08-3.98 (m, 1H), 3.65 (s, 3H), 3.62-3.53 (m, 2H), 2.87-2.73 (m, 1H), 2.69 (dd, J = 6.4, 9.6 Hz, 2H), 2.42-2.30 (m, 1H), 2.27-2.15 (m, 1H), 1.95-1.72 (m, 1H), 1.02-0.87 (m, 2H), -0.02 - -0.11 (m, 9H).

[0370] Step 6: To a solution of compound 4-6 (150 mg, 306.33 μmol) in tetrahydrofuran (2 mL) was added sodium hydride (36.76 mg, 919.00 μmol, 60% purity) at 0 °C under nitrogen atmosphere. The reaction solution was stirred at 0 °C for 0.5 h. Then the reaction solution was warmed to 25 °C and stirred for 1 h. The reaction solution was cooled to 0 °C and quenched with 40 mL of water. The reaction mixture was extracted with ethyl acetate (40 mL x 3). The combined organic phase was washed with 80 mL of saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give the crude product. The crude product was purified by column chromatography on silica gel (petroleum ether: ethyl acetate = 2: 1-0: 1) to give compound 4-7. LCMS: 468.3 [M+H] + .

[0371] Step 7: A solution of compound 4-7 (50 mg, 106.92 μmol) in trifluoroacetic acid (1 mL) was stirred at 25 °C for 1 h under nitrogen atmosphere. The reaction solution was concentrated under reduced pressure to give compound 4-8. LCMS: 368.1 [M+H] + .

[0372] Step 8: Compound 4-8 (35 mg, crude) was added to ammonia water (2 mL) and stirred at 25 °C for 1 h under nitrogen atmosphere. The reaction solution was concentrated under reduced pressure to give the crude product. The crude product was separated by preparative reversed-phase column chromatography (column: Phenomenex Luna C18 150*25mm*10μm; mobile phase: H20 (0.225% formic acid)-acetonitrile; gradient (acetonitrile%): 10%-40%) to give compound 4. LCMS: 338.1 [M+H] + ; 1H NMR (400 MHz, DMSO-d6) δ = 11.76 (br s, 1H), 8.38 (s, 1H), 7.46 (t, J = 3.0 Hz, 1H), 6.70 (dd, J = 1.7, 3.2 Hz, 1H), 4.63 - 4.43 (m, 2H), 4.05 - 3.91 (m, 2H), 3.91 - 3.82 (m, 1H), 2.93 - 2.83 (m, 1H), 2.80 - 2.72 (m, 1H), 2.42 - 2.30 (m, 1H), 2.13 - 2.01 (m, 1H), 1.95 (br d, J = 13.4 Hz, 1H), 1.76 - 1.57 (m, 1H), 0.97 - 0.88 (m, 2H), 0.88 - 0.79 (m, 2H); SFC detection (Chiralcel OJ-3 50*4.6mm I.D., 3pm; mobile phase: A phase: supercritical CO2, B phase: isopropanol (0.05% diethylamine); gradient (B%): 5%-40%), the retention time of compound 4 was 1.376 min, and the chiral purity was 100%.

[0373] Example 5

[0374] Step 1: To a solution of compound 4-3 (90 mg, 224.12 pmol) in N,N- dimethylformamide (1.5 mL) was added iron triflate (11.27 mg, 22.41 pmol) and compound 5-1 (55.96 mg, 268.94 pmol) under nitrogen atmosphere, the reaction was warmed to 60 °C and stirred for 0.5 h. The reaction was cooled to room temperature, diluted with 60 mL of ethyl acetate, the organic phase was washed with water (60 mL x 3) and saturated brine (60 mL) successively, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to give a crude product, which was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 2: 1-1:2) to give compound 5-2. LCMS: 480.2 [M+H] + , 1 H NMR (400 MHz, CDCl3) δ = 8.94 (s, 1H), 7.53 (d, J = 3.6 Hz, 1H), 6.87 (d, J = 3.6 Hz, 1H), 5.83 (s, 2H), 4.98 - 4.71 (m, 1H), 4.50 - 4.31 (m, 1H), 4.20 - 4.14 (m, 1H), 4.11 - 4.02 (m, 1H), 3.65 - 3.44 (m, 2H), 2.87 - 2.75 (m, 1H), 2.72 (dd, J = 2.0, 5.9 Hz, 2H), 2.34 - 2.14 (m, 2H), 1.95 - 1.81 (m, 1H), 0.99 - 0.89 (m, 2H), -0.05 (s, 9H).

[0375] Step 2: A solution of compound 5-2 (90 mg, 187.67 pmol) in trifluoroacetic acid (2.5 mL) was stirred at 25 °C for 1 h under nitrogen atmosphere. The reaction was concentrated under reduced pressure to give compound 5-3 as a crude product. LCMS: 380.1 [M+H] + .

[0376] Step 3: A solution of compound 5-3 (70 mg, 184.53 pmol) in ammonia (2.5 mL) was stirred at 25 °C for 1 h under nitrogen atmosphere. The reaction was concentrated under reduced pressure to give a crude product, which was separated by preparative reverse phase column chromatography (column Phenomenex Luna C18 150*25mm*10pm; mobile phase: H20 (0.225% formic acid) - acetonitrile; gradient (acetonitrile %): 30% - 60%) to give compound 5. LCMS: 350.1 [M+H] + ; 1 HNMR (400 MHz, DMSO-d6) d = 12.32 (br s, 1H), 8.78 (s, 1H), 7.64 (t, J = 3.2 Hz, 1H), 6.98 (br d, J = 1.6 Hz, 1H), 4.74 - 4.54 (m, 1H), 4.37 - 4.20 (m, 1H), 4.19 - 4.04 (m, 2H), 3.01 - 2.87 (m, 1H), 2.86 - 2.76 (m, 1H), 2.75 - 2.59 (m, 1H), 2.18 (br d, J = 12.6 Hz, 1H), 2.08 (br d, J = 13.7 Hz, 1H), 1.83 - 1.62 (m, 1H); 19 F NMR (376 MHz, DMSO-d6) d = -59.898; SFC detection (column Chiralpak IC-3 50*4.6 mm I.D., 3 pm; mobile phase: A phase is supercritical CO2, B phase is isopropanol / acetonitrile (0.05% diethylamine); gradient (B%): 20% - 60%), the retention time of compound 5 was 0.634 min with 100% chiral purity.

[0377] Example 6

[0378] Step 1: To a solution of compound 4-3 (1.2 g, 2.99 mmol) in toluene (15 mL) was added 1,1-carbonyldiimidazole (969 mg, 5.98 mmol) under nitrogen atmosphere. The reaction was heated to 60 °C and stirred for 2 h. The reaction was cooled to room temperature, diluted with 120 mL of ethyl acetate, washed with water (100 mL x 3) and saturated brine (120 mL) successively, dried over anhydrous sodium sulfate, filtered and the filtrate was concentrated under reduced pressure to give a crude product which was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 5:1-2:1) to give compound 6-1. LCMS: 428.1 [M+H] + ; 1 H NMR (400 MHz, CDCl3) d = 8.16 (s, 1H), 7.42 (d, J = 3.6 Hz, 1H), 6.69 (d, J = 3.6 Hz, 1H), 5.71 (s, 2H), 4.78-4.60 (m, 1H), 4.40 (br t, J = 11.2 Hz, 1H), 4.21-4.09 (m, 1H), 3.96-3.83 (m, 1H), 3.65-3.51 (m, 2H), 2.83 (dq, J = 4.0, 12.8 Hz, 1H), 2.67 (d, J = 5.6 Hz, 2H), 2.29-2.17 (m, 1H), 2.11 (br d, J = 12.4 Hz, 1H), 1.86-1.77 (m, 1H), 1.00-0.86 (m, 2H), -0.05 (s, 9H).

[0379] Step 2: To a solution of compound 6-1 (600 mg, 1.40 mmol) in N,N-dimethylformamide (7 mL) was added sodium hydride (171 mg, 4.29 mmol, 60% purity) under nitrogen atmosphere at 0 °C. The reaction was stirred at 0 °C for 0.5 h. CF2Br2 (1.96 g, 7.02 mmol) was added to the reaction, which was stirred at 0 °C for 2 h. The reaction was quenched by the addition of aqueous ammonium chloride solution (30 mL), extracted with ethyl acetate (60 mL x 3), washed with 50 mL of saturated brine, dried over anhydrous sodium sulfate, filtered and the filtrate was concentrated under reduced pressure to give a crude product which was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 10:1-0:1) and then by SFC (chromatographic column DAICEL CHIRALCEL OD (250 mm x 30 mm, 10 pm); mobile phase: A phase was supercritical CO2, B phase was ethanol (0.1% NH3H2O); gradient (B%): 25%) to give compound 6-2. LCMS: 556.0, 558.1 [M+H] + ; 1H NMR (400 MHz, CDC13) δ = 8.84 (s, 1H), 7.56-7.51 (m, 1H), 6.82 (d, J = 3.6 Hz, 1H), 5.84 (s, 2H), 4.98-4.76 (m, 1H), 4.32-4.10 (m, 2H), 3.97 (td, J = 5.2, 10.4 Hz, 1H), 3.67-3.61 (m, 2H), 2.76 (d, J = 5.6 Hz, 2H), 2.62 (dq, J = 4.0, 12.8 Hz, 1H), 2.46-2.34 (m, 1H), 2.23 (br d, J = 14.4 Hz, 1H), 1.91 (dq, J = 4.0, 12.8 Hz, 1H), 1.03-0.97 (m, 2H), 0.01 (s, 9H).

[0380] Step 3: To a solution of compound 6-2 (58 mg, 104 μmol) in dichloromethane (1 mL) was added silver tetrafluoroborate (153 mg, 784 μmol) in portions at room temperature under nitrogen atmosphere. The reaction was stirred at 25 °C for 22 h. The reaction was filtered, and the filtrate was purified by preparative reverse phase column chromatography (column: Phenomenex luna C18 (250 x 70 mm, 10 μm); mobile phase: water (0.1% formic acid) - acetonitrile; gradient (acetonitrile %): 55% - 100%) to give compound 6-3. LCMS: 496.2 [M+H] + .

[0381] Step 4: To a solution of compound 5 (10 mg, 20.18 μmol) in dichloromethane (0.3 mL) was added trifluoroacetic acid (0.15 mL) at room temperature under nitrogen atmosphere. The reaction was stirred at 25 °C for 12 h. The reaction was concentrated under reduced pressure to give compound 6-4 as a crude. LCMS: 396.1 [M+H] + .

[0382] Step 5: To a solution of compound 6-4 (7 mg, crude) in tetrahydrofuran (0.3 mL) was added ammonia water (20.7 μL, concentration 30%) at room temperature under nitrogen atmosphere. The reaction was stirred at 25 °C for 8 h. The reaction was filtered, and the filtrate was purified by preparative reverse phase column chromatography (column: Phenomenex luna C18 150 x 25 mm x 10 μm; mobile phase: H20 (0.225% formic acid) - acetonitrile]; gradient (acetonitrile %): 20% - 50%) to give compound 6. LCMS: 366.1 [M+H] + ; 1H NMR (400 MHz, CDC13) δ = 10.49-10.00 (m, 1H), 8.77-8.56 (m, 1H), 7.60-7.41 (m, 1H), 6.81 (s, 1H), 4.88-4.76 (m, 1H), 4.26-4.19 (m, 1H), 4.17-4.10 (m, 1H), 3.97-3.90 (m, 1H), 2.71 (d, J = 5.2 Hz, 2H), 2.61-2.51 (m, 1H), 2.38-2.31 (m, 1H), 2.19 (br d, J = 13.6 Hz, 1H), 1.90 (s, 1H); 19 F NMR (376 MHz, DMSO-d6) δ = -57.80.

[0383] Example 7

[0384] Step 1: Compound 3-2 (5.05 g, 20.91 mmol) was added to a solution of compound 7-1 (4.5 g, 20.91 mmol) in N,N-dimethylacetamide (50 mL) under nitrogen atmosphere, and reacted at 70 °C for 12 hours. The reaction solution was cooled to room temperature, 200 mL of water was added, and extracted with ethyl acetate (200 mL x 2). The combined organic phase was washed with 200 mL of saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain a crude product, which was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 10:1-3:1) to obtain compound 7-2. 1 H NMR (400 MHz, DMSO-d6) δ = 7.11 (br d, J = 5.8 Hz, 1H), 4.94-4.64 (m, 1H), 4.11 (br d, J = 6.5 Hz, 1H), 3.87-3.55 (m, 2H), 2.79-2.55 (m, 1H), 1.95 (br d, J = 6.5 Hz, 1H), 1.78-1.50 (m, 1H), 1.39 (d, J = 1.9 Hz, 9H).

[0385] Step 2: To a solution of compound 7-2 (4.5 g, 18.89 mmol) in methanol (50 mL) was added palladium on carbon (2.01 g, 5%) under nitrogen atmosphere, and after hydrogen replacement for three times, the reaction was carried out at 25 °C for 5 hours under hydrogen atmosphere (15 Psi). The reaction solution was filtered, and the filtrate was concentrated under reduced pressure to obtain compound 7-3. 1H NMR (400 MHz, DMSO-d6) δ = 6.89 - 6.31 (m, 1H), 3.95 - 3.68 (m, 1H), 3.58 (s, 2H), 3.32 - 3.12 (m, 1H), 2.81 - 2.55 (m, 2H), 1.92 - 1.55 (m, 2H), 1.54 - 1.42 (m, 1H), 1.39 (d, J = 8.9 Hz, 9H), 1.36 - 1.21 (m, 1H).

[0386] Step 3: To a solution of compound 7-3 (5.02 g, 20.91 mmol) in dichloromethane (50 mL) was added trimethylsilyl iodide (5.05 g, 25.25 mmol) at 0 °C under nitrogen atmosphere. The reaction was stirred at 0 °C for 1 h. The reaction was filtered and the filtrate was concentrated under reduced pressure to give the crude hydroiodide salt of compound 7-4.

[0387] Step 4: To a solution of compound 7-4 (3.1 g, crude hydroiodide salt) in tert-butanol (30 mL) was added compound 1-1 (4.07 g, 11.56 mmol) and N,N-diisopropylethylamine (7.47 g, 57.82 mmol) at 90 °C under nitrogen atmosphere. The reaction was stirred at 90 °C for 2.5 h. The reaction was cooled to room temperature, filtered and the filtrate was concentrated under reduced pressure to give the crude product which was purified by column chromatography on silica gel (petroleum ether: ethyl acetate = 10: 1-1: 1) to give compound 7-5A (petroleum ether: ethyl acetate = 1: 1, Rf = 0.45) and compound 7-5B (petroleum ether: ethyl acetate = 1: 1, Rf = 0.4). f f

[0388] Characterization of compound 7-5A: 1 H NMR (400 MHz, DMSO-d6) δ = 8.89 (s, 1H), 8.62 (d, J = 8.4 Hz, 1H), 8.00 (d, J = 8.4 Hz, 2H), 7.85 (d, J = 4.0 Hz, 1H), 7.44 (d, J = 8.4 Hz, 2H), 6.96 (d, J = 4.4 Hz, 1H), 4.19 - 4.04 (m, 2H), 3.74 - 3.58 (m, 1H), 3.44 (s, 1H), 2.95 - 2.65 (m, 2H), 2.36 (s, 3H), 2.17 (br d, J = 12.0 Hz, 1H), 1.80 (br d, J = 3.2 Hz, 2H), 1.66 - 1.48 (m, 1H).

[0389] Characterization of compound 7-5B: 1 ​​H NMR (400 MHz, DMSO-d6) δ = 9.40 (d, J = 8.0 Hz, 1H), 8.93 (s, 1H), 8.00 (d, J = 8.4 Hz, 2H), 7.80 (d, J = 4.0 Hz, 1H), 7.44 (d, J = 8.4 Hz, 2H), 7.02 (d, J = 4.4 Hz, 1H), 4.35 (br d, J = 7.6 Hz, 1H), 3.98 (br d, J = 12.6 Hz, 1H), 3.79 (d, J = 10.8 Hz, 1H), 3.76 - 3.66 (m, 1H), 2.87 - 2.62 (m, 2H), 2.36 (s, 3H), 1.95 (br s, 2H), 1.57 (br d, J = 1.6 Hz, 1H), 1.52 - 1.40 (m, 1H).

[0390] Step 5: To a solution of compound 7-5A (1.4 g, 3.07 mmol) in methanol (15 mL) was added palladium on carbon (1.31 g, 5% content) under nitrogen atmosphere, after three times of hydrogen replacement, stirred at 25 °C under hydrogen atmosphere (15 Psi) for 2 hours. The reaction solution was filtered, the filtrate was concentrated under reduced pressure to give compound 7-6A. 1 H NMR (400 MHz, DMSO-d6) δ = 7.90 (d, J = 8.4 Hz, 2H), 7.58 (s, 1H), 7.53 (d, J = 4.4 Hz, 1H), 7.38 (d, J = 8.4 Hz, 2H), 6.69 (d, J = 4.4 Hz, 1H), 5.09 (d, J = 8.8 Hz, 1H), 4.40 (s, 2H), 4.06 - 3.93 (m, 1H), 3.90 - 3.73 (m, 1H), 3.66 - 3.56 (m, 1H), 3.20 (s, 1H), 2.92 - 2.67 (m, 2H), 2.34 (s, 3H), 2.08 (br d, J = 12.0 Hz, 1H), 1.88 - 1.76 (m, 1H), 1.66 - 1.43 (m, 2H).

[0391] Step 6: To a solution of compound 7-6A (150 mg, 352.52 umol) and tetramethylammonium formate (479.95 mg, 3.53 mmol) in acetic acid (2 mL) was added p-toluenesulfonic acid monohydrate (6.71 mg, 35.25 umol) under nitrogen atmosphere, the reaction solution was stirred at 25 °C for 12 hours. The reaction solution was concentrated to give the crude product, which was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 100: 1-1: 2) to give compound 7-7A. LCMS: 466.1 [M+1] + .

[0392] Step 7: To a solution of compound 7-7A (110 mg, 236.29 pmol) in methanol (4 mL) was added potassium hydroxide aqueous solution (2 mL, 1 M) under nitrogen atmosphere. The reaction was heated to 60 °C for 4 h. The reaction was adjusted to pH = 7-7.5 with 1 M diluted hydrochloric acid and concentrated under reduced pressure to give the crude product, which was separated by preparative reverse phase chromatography (column: Waters Xbridge C18 150*25 mm*5 pm; mobile phase: H20 (0.05% NH3H20)-acetonitrile; gradient (acetonitrile %): 6%-36%) to give compound 7A. LCMS: 312.2 [M+1] + ; 1 H NMR (400 MHz, CDC13) d = 10.59 (br s, 1H), 8.60 (s, 1H), 7.42 (d, J = 2.0 Hz, 1H), 6.71 (d, J = 3.2 Hz, 1H), 4.90-4.63 (m, 1H), 4.26 (s, 3H), 4.23-4.07 (m, 2H), 3.90 (dtd, J = 1.6, 5.6, 11.2 Hz, 1H), 2.69 (d, J = 6.0 Hz, 2H), 2.66-2.48 (m, 1H), 2.31-2.21 (m, 1H), 2.12 (br s, 1H), 1.84 (br s, 1H); SFC detection (column: Chiralcel OD-3 50*4.6 mm I.D., 3 pm; mobile phase: A phase: supercritical CO2, B phase: isopropanol (0.05% diethylamine); gradient (B%): 5%-40%), the retention time of compound 7A was 1.636 min with a chiral purity of 98.77%.

[0393] Step 8: Refer to steps 5-7 above, compound 7B was prepared from compound 7-5B. LCMS: 312.2 [M+1] + .

[0394] Example 8

[0395] Step 1: To a mixture of trimethyl orthoacetate (84.71 mg, 705.05 pmol) and compound 7-6A (150 mg, 352.52 pmol) in toluene (1 mL) and tetrahydrofuran (1 mL) was added p-toluenesulfonic acid monohydrate (6.07 mg, 35.25 pmol) under nitrogen atmosphere. The reaction was heated to 100 °C for 3 h. The reaction was cooled to room temperature and concentrated under reduced pressure to give the crude product, which was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 10: 1-0: 1) to give compound 8-1A. LCMS: 450.1 [M+1] + .

[0396] Step 2: To a solution of compound 8-1A (60 mg, 133.47 pmol) in methanol (1 mL) was added potassium hydroxide aqueous solution (1.20 mL, 1.0 M) under nitrogen atmosphere. The reaction was stirred at 25 °C for 12 h. The reaction was adjusted to pH = 7-7.5 with 1 M diluted hydrochloric acid, and concentrated under reduced pressure to give the crude product, which was separated by preparative reverse phase chromatography (column: Waters Xbridge C18 150*25mm*5pm; mobile phase: H20 (0.05% NH3-H20) - acetonitrile; gradient (B%): 5% - 35%) to give compound 8A. LCMS: 296.2 [M+1] + ; 1 H NMR (400 MHz, DMSO-d6) d = 11.89 (br s, 1H), 8.49 (s, 1H), 7.50 (d, J = 3.2 Hz, 1H), 6.82 (d, J = 3.2 Hz, 1H), 4.71 - 4.55 (m, 1H), 4.23 - 4.01 (m, 3H), 2.99 - 2.77 (m, 2H), 2.68 (s, 3H), 2.65 - 2.54 (m, 1H), 2.14 (br d, J = 12.0 Hz, 1H), 2.04 (br d, J = 13.6 Hz, 1H), 1.75 (dq, J = 4.0, 12.8 Hz, 1H). SFC detection (column: Chiralpak AD-3 50*4.6mm I.D., 3pm; mobile phase: A phase: supercritical CO2, B phase: isopropanol + acetonitrile (0.05% diethylamine); gradient (B%): 20% - 60%) the retention time of compound 8A was 1.176 min with 98.83% chiral purity.

[0397] Step 3: Refer to Step 1, compound 8-1B was prepared from compound 7-6B. To a solution of compound 8-1B (20 mg, 43.38 pmol) in methanol (1 mL) was added magnesium powder (21.09 mg, 867.58 pmol) under nitrogen atmosphere. The reaction was stirred at 25 °C for 2 h. The reaction was filtered, and the filtrate was concentrated under reduced pressure to give the crude product, which was separated by preparative reverse phase chromatography (column: Waters Xbridge C18 150*25mm*5pm; mobile phase: H20 (0.05% NH3-H20) - acetonitrile; gradient (acetonitrile%): 5% - 35%) to give compound 8B. LCMS: 296.1 [M+1] + ; 1H NMR (400 MHz, DMSO-d6) δ = 11.80 (br s, 1H), 8.46 (s, 1H), 7.46 (d, J = 3.2 Hz, 1H), 6.73 (d, J = 3.2 Hz, 1H), 4.95 - 4.84 (m, 1H), 4.45 - 4.37 (m, 1H), 4.14 - 3.97 (m, 2H), 3.07 - 2.90 (m, 2H), 2.77 (s, 3H), 2.37 - 2.26 (m, 1H), 2.24 - 2.12 (m, 1H), 1.92 - 1.80 (m, 1H), 1.67 - 1.53 (m, 1H). SFC detection (Chiralpak AD-35 0*4.6 mm I.D., 3 pm; mobile phase: A phase: supercritical CO2, B phase: isopropanol + acetonitrile (0.05% diethylamine); gradient (B%): 20% - 60%) Retention time of compound 8B was 1.422 min with 99.54% chiral purity.

[0398] Example 9

[0399] Following the synthetic procedure of the above example, compound 9A was prepared as a crude product which was purified by preparative reverse phase chromatography (Chromolith® column Phenomenex Luna C18 150*25 mm*10 pm; mobile phase: H20 (0.225% formic acid) - acetonitrile; gradient (acetonitrile%): 30% - 50%) to give 9A. LCMS: 350.1 [M+1] + ; 1 H NMR (400 MHz, DMSO-d6) δ = 12.32 (br s, 1H), 8.79 (s, 1H), 7.65 (t, J = 3.0 Hz, 1H), 6.99 (br s, 1H), 4.65 (br s, 1H), 4.31 - 4.21 (m, 1H), 4.19 - 3.95 (m, 2H), 2.98 - 2.89 (m, 1H), 2.87 - 2.76 (m, 1H), 2.75 - 2.58 (m, 1H), 2.18 (br d, J = 11.5 Hz, 1H), 2.08 (br d, J = 13.6 Hz, 1H), 1.81 - 1.61 (m, 1H); 19 F NMR (376 MHz, DMSO-d6) δ = 59.91. SFC detection (Chiralpak AD-3 50*4.6 mm I.D., 3 pm; mobile phase: A phase: supercritical CO2, B phase: ethanol (0.05% diethylamine); gradient (B%): 10% - 60%) Retention time of compound 9A was 0.624 min with 100% chiral purity.

[0400] Example 10

[0401] Step 1: Refer to the above example, compound 2-1A was used as raw material to prepare compound 2-7A.

[0402] Step 2: To a solution of compound 2-7A (1 g, 3.74 mmol) and triethylamine (2.08 mL, 15.0 mmol) in dichloromethane (10 mL) was added methanesulfonyl chloride (790 μL, 10.2 mmol) at 0 °C under nitrogen atmosphere. The reaction was warmed to 25 °C and stirred for 1 h. To the reaction was added 50 mL water and extracted with ethyl acetate (100 mL x 2). The combined organic phase was washed with 100 mL saturated brine, dried over anhydrous sodium sulfate, filtered and the filtrate was concentrated under reduced pressure to give compound 10-2.

[0403] Step 3: To a solution of compound 10-2 (1.29 g, 3.73 mmol) in N,N-dimethylacetamide (10 mL) was added sodium cyanide (810 mg, 16.5 mmol) at 25 °C under nitrogen atmosphere. The reaction was stirred for 1 h. To the reaction was added 50 mL water and extracted with ethyl acetate (100 mL x 2). The combined organic phase was washed with 100 mL saturated brine, dried over anhydrous sodium sulfate, filtered and the filtrate was concentrated under reduced pressure to give the crude product which was purified by column chromatography on silica gel (petroleum ether / ethyl acetate = 10 / 1 to 0 / 1) to give compound 10-3. LCMS: 221.1 [M-56+H] + .

[0404] Step 4: To a solution of compound 10-3 (100 mg, 362 μmol) in dichloromethane (1 mL) was added trimethylsilyl iodide (59.1 μL, 434 μmol) slowly at 0 °C under nitrogen atmosphere. The reaction was stirred at 0 °C for 0.5 h. The reaction was concentrated under reduced pressure to give the crude hydroiodide salt of compound 10-4.

[0405] Step 5: To a solution of compound 10-4 (100 mg, crude hydroiodide salt) and compound 2-12 (78.4 mg, 395 μmol) in isopropanol (2 mL) was added N,N-diisopropylethylamine (286 μL, 1.64 mmol) at 80 °C under nitrogen atmosphere. The reaction was stirred at 80 °C for 12 h. The reaction was cooled to room temperature, diluted with 5 mL water and extracted with ethyl acetate (100 mL x 2). The combined organic phase was washed with 100 mL saturated brine, dried over anhydrous sodium sulfate, filtered and the filtrate was concentrated under reduced pressure to give the crude product which was purified by column chromatography on silica gel (petroleum ether / ethyl acetate = 10 / 1 to 0 / 1) to give compound 10-5. 1H NMR (400 MHz, DMSO-d6) δ = 9.08 (s, 1H), 8.62 (d, J = 8.8 Hz, 1H), 8.45 (d, J = 2.0 Hz, 1H), 7.17 (d, J = 2.0 Hz, 1H), 6.07 (s, 1H), 5.14-4.95 (m, 1H), 4.50 (dd, J = 4.8, 12.4 Hz, 1H), 4.23 (dd, J = 8.0, 12.4 Hz, 1H), 3.92 (s, 2H), 2.99-2.87 (m, 2H), 2.31-2.09 (m, 2H). SFC detection (Chromolith® Speed Ray® column Chiralcel OJ-3 50*4.6mm I.D., 3 μm, mobile phase: A phase: supercritical CO2, B phase: isopropanol (0.05% diethylamine); gradient (B%): 5%-40%) the retention time of compound 10-5 was 1.357 min with 99.43% chiral purity.

[0406] Step 6: To a mixture of compound 10-5 (100 mg, 295 μmol) in dichloromethane (2 mL) and methanol (2 mL) was added palladium on carbon (94.4 mg, 5% content) under nitrogen atmosphere, and the hydrogen gas was replaced for three times. The reaction mixture was stirred at 25 °C under hydrogen atmosphere (15 psi) for 0.5 h. The reaction mixture was filtered, and the filtrate was concentrated under reduced pressure to give compound 10-6. LCMS: 309.1 [M+1] + .

[0407] Step 7: To a mixture of compound 10-6 (90 mg, 292 μmol) in toluene (1 mL) and tetrahydrofuran (1 mL) was added trimethyl orthoacetate (70.1 mg, 584 μmol) and p-toluenesulfonic acid (5.03 mg, 29.2 μmol) sequentially under nitrogen atmosphere at room temperature. The reaction mixture was warmed to 70 °C and stirred for 5 h. The reaction mixture was cooled to room temperature, diluted with 5 mL of water, extracted with ethyl acetate (50 mL x 2), and the combined organic phase was washed with 50 mL of saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to give the crude product, which was purified by silica gel column chromatography (dichloromethane / methanol = 1 / 0-10 / 1) to give compound 10. LCMS: 333.1 [M+1] + ; 1H NMR (400 MHz, DMSO-d6) δ = 8.83 (s, 1H), 8.27 (d, J = 2.0 Hz, 1H), 7.23 (d, J = 2.0 Hz, 1H), 6.14 (s, 1H), 5.31-5.13 (m, 1H), 4.69 (dd, J = 6.0, 12.0 Hz, 1H), 4.56-4.43 (m, 1H), 3.96 (s, 2H), 3.17-2.99 (m, 2H), 2.75 (s, 3H), 2.73-2.64 (m, 1H), 2.34-2.23 (m, 1H). SFC detection (Chiralcel OJ-3 50*4.6mm I.D., 3pm, mobile phase: A phase: supercritical CO2, B phase: isopropanol (0.05% diethylamine); gradient (B%): 5%-40%) the retention time of compound 10 was 1.326 min with 100% chiral purity.

[0408] Example 11

[0409] Step 1: Compound 11-1 (9.20 g, 47.18 mmol) and compound 11-2 (6.56 g, 47.18 mmol) were stirred in ethylene glycol dimethyl ether (100 mL) at 25 °C for 1 h under nitrogen atmosphere. The reaction was concentrated under reduced pressure to remove ethylene glycol dimethyl ether, and the residue was dissolved in 100 mL of ethanol and heated to 80 °C for 8 h with stirring. The reaction was cooled to room temperature and concentrated under reduced pressure to give a crude product, which was purified by silica gel column chromatography (dichloromethane / methanol = 1 / 0 to 15 / 1) to give compound 11-3. LCMS: 236.1 [M+1] + ; 1 H NMR (400 MHz, DMSO-d6) δ = 8.83 (s, 1H), 8.76 (dd, J = 0.8, 7.6 Hz, 1H), 8.67 (d, J = 2.4 Hz, 1H), 7.76 (dd, J = 2.4, 7.6 Hz, 1H), 4.35 (q, J = 7.2 Hz, 2H), 1.34 (t, J = 7.2 Hz, 3H).

[0410] Step 2: To compound 11-3 (3.0 g, 12.76 mmol) in ethanol (200 mL) and aqueous hydrochloric acid (1 M, 20 mL) was added platinum dioxide (290 mg, 1.28 mmol) under nitrogen atmosphere. After hydrogen replacement for three times, the reaction was stirred at 50 °C under hydrogen atmosphere (50 psi) for 12 h. The reaction was cooled to room temperature, filtered, and the filtrate was concentrated under reduced pressure to give a crude hydrochloride salt of compound 11-4. LCMS: 210.1 [M+1] + .

[0411] Step 3: To a solution of compound 11-4 (2.4 g, crude hydrochloride salt) in dichloromethane (50 mL) was added triethylamine (3.48 g, 34.4 mmol) and di-tert-butyl dicarbonate (Boc anhydride, 3.75 g, 17.2 mmol) under nitrogen atmosphere at 25 °C and stirred for 2 h. To the reaction mixture was added water (80 mL) and extracted with dichloromethane (80 mL x 3), the combined organic phase was washed with saturated sodium chloride solution (200 mL), dried over anhydrous sodium sulfate, filtered and the filtrate was concentrated under reduced pressure to give a crude product which was purified by column chromatography on silica gel (dichloromethane / methanol = 1 / 0 to 15 / 1) to give compound 11-5. LCMS: 310.2 [M+1] + .

[0412] Step 4: To a solution of compound 11-5 (3.0 g, 9.70 mmol) in anhydrous tetrahydrofuran (30 mL) was added diisobutylaluminum hydride solution in hexane (1 M, 19.4 mL, 19.4 mmol) dropwise at -78 °C under nitrogen atmosphere and stirred for 4 h at -78 °C. To the reaction mixture was added 60 mL of water dropwise to quench the reaction and extracted with dichloromethane (60 mL x 3), the combined organic phase was washed with 120 mL of saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered and the filtrate was concentrated under reduced pressure to give a crude product which was purified by column chromatography on silica gel (dichloromethane / methanol = 1 / 0 to 10 / 1) to give compound 11-6. LCMS: 268.2 [M+1] + . 1 H NMR (400 MHz, CDCl3) d = 6.76 (s, 1H), 4.72 (br d, J = 7.1 Hz, 1H), 4.54 (s, 2H), 4.27-3.57 (m, 3H), 3.19 (br dd, J = 5.2, 16.6 Hz, 1H), 2.75-2.75 (m, 1H), 2.71 (dd, J = 7.9, 16.5 Hz, 1H), 2.31-2.19 (m, 1H), 1.45 (s, 9H).

[0413] Step 5: To a mixture of compound 11-6 (2 g, 7.48 mmol) in dichloromethane (20 mL) and methanol (20 mL) was added manganese dioxide (6.50 g, 74.8 mmol) portionwise slowly and the mixture was heated to 65 °C and stirred for 12 h. After completion of the reaction, the reaction mixture was allowed to cool to room temperature, filtered and the filtrate was concentrated under reduced pressure to give a crude product which was purified by column chromatography on silica gel (dichloromethane / methanol = 1 / 0 to 15 / 1) to give compound 11-7. 1H NMR (400 MHz, CDC13) δ = 9.83 (s, 1H), 7.53 (s, 1H), 4.63 (br d, J = 1.2 Hz, 1H), 4.24-4.00 (m, 3H), 3.29 (dd, J = 5.2, 16.8 Hz, 1H), 2.79 (dd, J = 8.4, 16.8 Hz, 1H), 2.39-2.25 (m, 1H), 2.16-2.03 (m, 1H), 1.46 (s, 9H).

[0414] Step 6: To a solution of compound 11-7 (900 mg, 3.39 mmol) in methanol (5 mL) was added sodium acetate (417 mg, 5.09 mmol) and hydroxylamine hydrochloride (283 mg, 4.07 mmol) under nitrogen atmosphere. The reaction was stirred at 25 °C for 1 h. The reaction was diluted with 20 mL water and extracted with ethyl acetate (20 mL x 2). The combined organic phase was washed with 20 mL saturated brine, dried over sodium sulfate, filtered and the filtrate was concentrated under reduced pressure to give compound 11-8. LCMS: 281.2 [M+1] + .

[0415] Step 7: To a solution of compound 11-8 (900 mg, 3.21 mmol) in tetrahydrofuran (5 mL) was added slowly with Burgess reagent (1.15 g, 4.82 mmol) at 0 °C under nitrogen atmosphere. The reaction was stirred at 25 °C for 12 h. The reaction was quenched with 5 mL water and extracted with ethyl acetate (20 mL x 2). The combined organic phase was washed with 30 mL saturated brine, dried over sodium sulfate, filtered and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 20 / 1 to 1 / 1) to give compound 11-9. 1 H NMR (400 MHz, CDC13) δ = 7.37 (s, 1H), 4.79 (br d, J = 6.8 Hz, 1H), 4.19-3.97 (m, 3H), 3.23 (dd, J = 5.2, 16.8 Hz, 1H), 2.75 (dd, J = 8.0, 16.8 Hz, 1H), 2.36-2.21 (m, 1H), 2.18-2.06 (m, 1H), 1.46 (s, 9H).

[0416] Step 8: To a solution of compound 11-9 (180 mg, 686 μmol) in dichloromethane (1 mL) was added slowly with trimethylsilyl iodide (112 μL, 823 μmol) at 0 °C under nitrogen atmosphere. The reaction was stirred at 0 °C for 0.5 h. The reaction was concentrated to give the crude hydroiodide salt of compound 11-10.

[0417] Step 9: To a solution of compound 11-10 (180 mg, crude hydroiodide) and compound 2-12 (148 mg, 745 pmol) in isopropanol (2 mL) was added N,N- diisopropylethylamine (401 mg, 3.10 mmol) under nitrogen atmosphere. The reaction was heated to 80 °C and stirred for 12 h. The reaction was cooled to room temperature, diluted with 10 mL of water and extracted with ethyl acetate (20 mL x 2). The combined organic phase was washed with 50 mL of saturated brine, dried over anhydrous sodium sulfate, filtered and the filtrate was concentrated under reduced pressure to give the crude product which was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 10 / 1 to 0 / 1) to give compound 11-11.

[0418] Step 10: To a solution of compound 11-11 (180 mg, 555 pmol) in dichloromethane (5 mL) and methanol (5 mL) was added palladium on carbon (100 mg, 5% loading) under nitrogen atmosphere. The reaction was purged with hydrogen three times and stirred under hydrogen atmosphere (15 psi) at 25 °C for 0.5 h. The reaction was filtered and the filtrate was concentrated under reduced pressure to give compound 11-12. LCMS: 295.2 [M+1] + .

[0419] Step 11: To a solution of compound 11-12 (160 mg, 544 pmol) in toluene (2 mL) and tetrahydrofuran (2 mL) was added trimethyl orthoacetate (131 mg, 1.09 mmol) and p-toluenesulfonic acid (9.36 mg, 54.4 pmol) sequentially under nitrogen atmosphere at room temperature. The reaction was heated to 70 °C and stirred for 5 h. The reaction was cooled to room temperature, diluted with 10 mL of water and extracted with ethyl acetate (20 mL x 2). The combined organic phase was washed with 50 mL of saturated brine, dried over anhydrous sodium sulfate, filtered and the filtrate was concentrated under reduced pressure to give the crude product which was purified by silica gel column chromatography (dichloromethane / methanol = 1 / 0 to 10 / 1) to give compound 11. LCMS: 319.1 [M+1] + .

[0420] Step 12: Compound 11 was separated by SFC (Chromatography column: DAICEL CHIRALPAK AD (250 mm x 30 mm, 10 pm, mobile phase: A phase: supercritical CO2, B phase: ethanol / acetonitrile (4: 1) (0.1% NH3H2O); gradient (B%): 60%) to give compound 11A and compound 11B.

[0421] Characterization of compound 11A: LCMS: 319.1 [M+1] + ; 1H NMR (400 MHz, DMSO-d6) δ = 8.83 (s, 1H), 8.25 (d, J = 2.4 Hz, 1H), 8.16 (s, 1H), 7.23 (d, J = 2.4 Hz, 1H), 5.32-5.16 (m, 1H), 4.42 (br dd, J = 4.8, 12.8 Hz, 1H), 4.26 (dt, J = 4.4, 12.8 Hz, 1H), 3.51-3.43 (m, 2H), 2.93-2.81 (m, 1H), 2.71 (s, 3H), 2.39 (br d, J = 12.8 Hz, 1H). SFC detection (Chiralpak AD-3 50 x 4.6 mm I.D., 3 pm, mobile phase: A phase supercritical CO2, B phase ethanol (0.05% diethylamine); gradient (B%): 30%-60%), the retention time of compound 11A was 0.829 min with 100% chiral purity.

[0422] Compound 11B characterization: LCMS: 319.1 [M+1] + ; 1 H NMR (400 MHz, DMSO-d6) δ = 8.83 (s, 1H), 8.25 (d, J = 2.4 Hz, 1H), 8.16 (s, 1H), 7.23 (d, J = 2.4 Hz, 1H), 5.32-5.16 (m, 1H), 4.42 (br dd, J = 4.8, 12.8 Hz, 1H), 4.26 (dt, J = 4.4, 12.8 Hz, 1H), 3.51-3.43 (m, 2H), 2.93-2.81 (m, 1H), 2.71 (s, 3H), 2.39 (br d, J = 12.8 Hz, 1H). SFC detection (Chiralpak AD-3 50 x 4.6 mm I.D., 3 pm, mobile phase: A phase supercritical CO2, B phase ethanol (0.05% diethylamine); gradient (B%): 30%-60%), the retention time of compound 11A was 0.829 min with 100% chiral purity.

[0423] Example 12

[0424] To a solution of compound 10 (29 mg, 87.26 μmol) in N,N-dimethylformamide (1 mL) was added 1.8-diazabicyclo[5.4.0]undec-7-ene (26.57 mg, 174.51 μmol) and iodomethane (18.58 mg, 130.88 μmol) under nitrogen atmosphere. The reaction was stirred at 60 °C for 1 h. The reaction was filtered and the filtrate was concentrated under reduced pressure to give a crude product, which was purified by preparative reverse phase chromatography (column: Phenomenex Luna C18 150*25mm*10μm; mobile phase: H2O (0.225% formic acid) - acetonitrile; gradient (acetonitrile %): 0% - 30%) to give compound 12. LCMS: 347.3. [M+H] + ; 1 H NMR (400 MHz, DMSO-d6) δ = 9.65 (s, 1H), 8.75 (d, J = 2.0 Hz, 1H), 8.51 (s, 1H), 7.86 (d, J = 2.0 Hz, 1H), 6.15 (s, 1H), 5.48-5.29 (m, 1H), 4.76 (dd, J = 6.0, 12.1 Hz, 1H), 4.58-4.41 (m, 4H), 3.97 (s, 2H), 3.18-3.02 (m, 2H), 2.90 (s, 3H), 2.76-2.60 (m, 1H), 2.45-2.34 (m, 1H).

[0425] Example 13: Preparation of the A crystal form of compound 3

[0426] Compound 3 (100 mg) was added to n-heptane (1 mL), and the suspension was stirred at 50 °C or room temperature for 3 days, and then dried under vacuum at 40 °C overnight to give the A crystal form of compound 3, whose XRPD, DSC, and TGA detection results are shown in Figures 3, 4, and 5, respectively.

[0427] Example 14: Preparation of the B crystal form of compound 3

[0428] Method 1: Compound 3 (10 mg) was added to solvent X (solvent X is 0.5 mL of ethanol), and stirred at 50 °C for 3 days, and then dried under vacuum at 40 °C overnight to give the B crystal form of compound 3, whose XRPD, DSC, and TGA detection results are shown in Figures 6, 7, and 8, respectively.

[0429] The above solvent X can also be selected from ethanol / water (9 / 1, v / v), or the solvent X is a mixed solvent selected from one or more of isopropanol, acetone, acetonitrile, ethyl acetate, tetrahydrofuran, methyl tert-butyl ether, 2-methyltetrahydrofuran, isopropyl acetate, dichloromethane, toluene, 1,4-dioxane.

[0430] Method 2: Compound 3 (10 mg) was added into solvent Y (0.5 mL), stirred at room temperature for 3 days, and then dried at 40 °C under vacuum overnight to obtain the B crystal form of compound 3. Wherein, solvent Y is selected from ethanol / water (9 / 1, v / v), or solvent Y is selected from a mixed solvent of one or more of ethanol, isopropanol, acetone, acetonitrile, tetrahydrofuran, 2-methyltetrahydrofuran, isopropyl acetate, dichloromethane, 1,4-dioxane.

[0431] Method 3: Compound 3 (50 mg) was added into solvent Z (0.5-1 mL), and then subjected to temperature cycling (5-55 °C, heating rate of 1 °C; 55-5 °C, cooling rate of 0.25 °C / min, 13 cycles), and then dried at 40 °C under vacuum overnight to obtain the B crystal form of compound 3. Wherein, solvent Z is selected from ethanol / water (9 / 1, v / v), or solvent Z is selected from a mixed solvent of one or more of ethanol, isopropanol, acetone, acetonitrile, tetrahydrofuran, 1,4-dioxane.

[0432] Example 15: Preparation of the C crystal form of compound 3

[0433] Compound 3 (100 mg) was added into methanol (0.5 mL), stirred at room temperature for 3 days, and then dried at 40 °C under vacuum overnight to obtain the C crystal form of compound 3, and the XRPD, DSC, and TGA detection results thereof are shown in FIGS. 9, 10, and 11, respectively.

[0434] Example 16: Preparation of the D crystal form of compound 3'

[0435] Compound 3 (100 mg) was added into water (2 mL), stirred at room temperature for 3 days, and then dried at 40 °C under vacuum overnight to obtain the D crystal form of compound 3', and the XRPD, DSC, and TGA detection results thereof are shown in FIGS. 12, 13, and 14, respectively.

[0436] Example 17: Solid stability study

[0437] The B crystal form of compound 3 was stored at 40 °C / 75% RH, 25 °C / 60% RH, high temperature 60 °C, high humidity 90% RH, and light conditions (total illumination not less than 1.2 x 10 6 Lux·hr, near ultraviolet energy not less than 200 w·hr / m 2 under a light source) for 1 month, and the XRPD pattern showed that the crystal form did not change.

[0438] The D crystal form of compound 3' was stored at 40 °C / 75% RH, 25 °C / 60% RH, high temperature 60 °C, and light conditions (total illumination not less than 1.2 x 10 6 Lux·hr, near ultraviolet energy not less than 200 w·hr / m 2XRPD pattern showed no change in crystal form after 1 month storage under light at 40 °C / 75% RH.

[0439] The B form of compound 3 and the D form of compound 3' have good solid stability under high temperature, high humidity or light conditions.

[0440] Example 18: Hygroscopicity study

[0441] DVS test conditions are shown in Table 6. DVS test results showed that the hygroscopic weight gain of the B form of compound 3 under 80% RH was 0.2314%, and the DVS spectrum of the B form of compound 3 is shown in Figure 15; the hygroscopic weight gain of the D form of compound 3' under 80% RH was 1.536%, and the DVS spectrum of the D form of compound 3' is shown in Figure 16.

[0442] Hygroscopicity evaluation classification: dissolution: absorption of sufficient amount of water to form a liquid; very hygroscopic: AW% ≥ 15%; hygroscopic: 15% > AW% ≥ 2%; slightly hygroscopic: 2% > AW% ≥ 0.2%; no or almost no hygroscopicity: AW% < 0.2%.

[0443] Conclusion: the B form of compound 3 and the D form of compound 3' are slightly hygroscopic under 80% RH.

[0444] Reference compound 1

[0445] Reference compound 1 is Example 11 of patent WO2018067422A1.

[0446] Reference compound 2

[0447] Reference compound 2 was prepared according to the preparation method of Example 457 / 458 of WO2011086053A1, and then reference compound 2A and reference compound 2B were obtained by SFC preparation separation (chromatographic column: DAICEL CHIRALPAK AD (250 mm*30 mm, 10 μm, mobile phase: A phase is supercritical CO2, B phase is methanol (0.1% ammonia water); gradient (B%): 30%).

[0448] Characterization of reference compound 2A: LCMS: 257.3 [M+H] + ; 1H NMR (400 MHz, DMSO-d6) δ = 11.85 (s, 1H), 8.47 (s, 1H), 7.48 (t, J = 2.8 Hz, 1H), 6.81 (s, 1H), 4.79-4.42 (s, 1H), 4.18-3.86 (m, 3H), 3.75-3.55 (m, 1H), 2.65 (s, 3H), 2.59-2.51 (m, 1H), 2.09 (d, J = 11.6 Hz, 1H), 1.89 (s, 2H); SFC detection (column: Chiralpak AD-3 50*4.6 mm I.D., 3 pm, mobile phase: A phase: supercritical CO2, B phase: methanol (0.05% diethylamine); gradient (B%): 5%-40%), retention time of reference compound 2A was 1.743 min, chiral purity was 100%.

[0449] Reference compound 2B characterization: LCMS: 257.3 [M+H] + ; 1 H NMR (400 MHz, DMSO-d6) δ = 11.85 (s, 1H), 8.47 (s, 1H), 7.48 (t, J = 2.8 Hz, 1H), 6.81 (s, 1H), 4.79-4.42 (s, 1H), 4.18-3.86 (m, 3H), 3.75-3.55 (m, 1H), 2.65 (s, 3H), 2.59-2.51 (m, 1H), 2.09 (d, J = 11.6 Hz, 1H), 1.89 (s, 2H); SFC detection (column: Chiralpak AD-3 50*4.6 mm I.D., 3 pm, mobile phase: A phase: supercritical CO2, B phase: methanol (0.05% diethylamine); gradient (B%): 5%-40%), retention time of reference compound 2B was 1.886 min, chiral purity was 97.86%.

[0450] Biological test data

[0451] Test Example 1: Kinase activity test

[0452] Enzyme activity detection experiment adopts homogeneous time-resolved fluorescence resonance energy transfer technology (HTRF) of Cisbio Company to detect the inhibition effect of compound on JAK1 / JAK2 / JAK3 / TYK2 JH1 kinase (JAKs) activity.

[0453] 1. Experimental materials:

[0454] JAK1, JAK2, JAK3, TYK2 kinase proteins are purchased from Thermo Fisher Company; KinEASE TM- TK 20000tests kit purchased from Cisbio; Microplate Multilabel Reader: PerkinElmer, Model Envision 2104 Multilabel Reader.

[0455] 2. Experimental Methods:

[0456] (1) Preparation of reaction buffer: 10 mL as an example, prepared on the experimental day. The preparation parameters are shown in Tables 7.1-7.3.

[0457] Table 7.1 JAK1 JH1 reaction buffer

[0458] Table 7.2 JAK2 / 3 JH1 reaction buffer

[0459] Table 7.3 TYK2 JH1 reaction buffer

[0460] (2) Experimental process

[0461] Step a, pre-incubation of compound and kinase: 2x JAK1 / JAK2 / JAK3 / TYK2 JH1 enzyme solution was prepared using the corresponding reaction buffer (enzyme final concentration was 0.133 ng / μL, 0.0225 ng / μL, 0.224 ng / μL, 0.25 ng / μL, respectively). The compound was transferred to the 384-well experimental plate (DMSO was used instead of negative and positive controls) by 100 nL per well using an automated micropipetting system, 5 μL of 2x JAK1 / JAK2 / JAK3 / TYK2 JH1 enzyme solution was added to each well (compound well and positive control well) or reaction buffer (negative control well), centrifuged and mixed, and incubated at 25°C for 15 minutes.

[0462] Step b, enzyme reaction: 2x TK-Sub-biotin substrate and ATP mixed solution was prepared using the corresponding reaction buffer (in JAK1, JAK2, JAK3, TYK2 enzyme reaction, the final concentration of TK-Sub-biotin substrate was 5 μM, 2 μM, 0.6 μM, 1 μM, respectively; the final concentration of ATP was 2 μM, 0.7 μM, 0.85 μM, 0.1 μM, respectively). After the completion of step a incubation, 5 μL of 2x TK-Sub-biotin substrate and ATP mixed solution was added to each well of the above 384-well experimental plate, centrifuged and mixed, and reacted at 25°C for 45 minutes (JAK1 / JAK2 JH1) or 60 minutes (JAK3 / TYK2 JH1).

[0463] Step c, detection: detection buffer was used to prepare the detection mixture of TK Antibody-Eu and streptavidin-XL665 (final concentration of TK Antibody-Eu was 0.25x, final concentration of streptavidin-XL665 was 1 / 16 of the final concentration of TK-Sub-biotin substrate). After the completion of step b incubation, 10 μL of the detection mixture was added to each well of the above-mentioned 384-well experimental plate, centrifuged and mixed, and then reacted at 25°C for 60 minutes (JAK1 / JAK2 JH1) or 120 minutes (JAK3 / TYK2 JH1). After the reaction was completed, the fluorescence value was read on Envision 2104 Multilabel Reader (340 nm excitation, detection of 665 nm and 615 nm emission, 665 nm / 615 nm fluorescence ratio was the original data of the well reaction signal value).

[0464] 3. Data processing and analysis

[0465] First, the average values of the reaction signals of the positive control wells and the negative control wells were calculated, respectively, and then the reaction inhibition rate of each compound well could be calculated according to the formula "single-well inhibition rate = (1-(single-well signal value-negative control signal average value) / (positive control signal average value-negative control signal average value))x100%". Then, the concentration and corresponding inhibition rate data were imported into XLfit software, and the Dose Response One Site205 model in the software was used to fit the four-parameter inhibition rate-concentration curve, and the IC 50 value of the compound was calculated.

[0466] The experimental results are shown in Table 7.4. Among them, A represents IC 50 ≤1nM (A+ further represents IC 50 ≤0.2nM), B represents 1nM<IC 50 ≤20nM (B+ further represents 1nM<IC 50 ≤10nM), C represents 20nM<IC 50 ≤100nM (C+ further represents 20nM<IC 50 ≤50nM), D represents 100nM<IC 50 ≤500nM (D+ further represents 100nM<IC 50 ≤200nM), E represents IC 50 >500nM. The JAK1-JAK2 selectivity factor is the ratio of the JAK2 IC 50 value to the JAK1 IC 50 value, and the JAK1-JAK3 selectivity factor is the ratio of the JAK3 IC 50 value to the JAK1 IC50 The ratio of the values.

[0467] The experimental results show that the compounds of the present application have strong inhibitory activity on JAK1 kinase, which is significantly better than that of the control compounds. Specifically, the IC 50 of compounds 4 and 5 are all ≤1 nM (5 times better than the inhibitory activity of the control compound 1, 17.5 times better than the inhibitory activity of the control compound 2A, and 55.3 times better than the inhibitory activity of the control compound 2B), and the IC 50 of compounds 1A, 3 and 6 are all ≤0.2 nM (10 times, 25 times and 9.5 times better than the inhibitory activity of the control compound 1, respectively, all more than 30 times better than the inhibitory activity of the control compound 2A, and all more than 100 times better than the inhibitory activity of the control compound 2B).

[0468] The compounds of the present application have strong inhibitory activity on TYK2 kinase, which is significantly better than that of the control compounds. Specifically, the IC 50 of compound 1A is ≤10 nM (15 times better than the inhibitory activity of the control compound 1, 17.2 times better than the inhibitory activity of the control compound 2A, and more than 30 times better than the inhibitory activity of the control compound 2B), and the IC 50 of compound 3 is ≤1 nM (15 times better than the inhibitory activity of the control compound 1, more than 100 times better than the inhibitory activity of the control compound 2A, and more than 100 times better than the inhibitory activity of the control compound 2B).

[0469] The inhibitory activity of the compounds of the present application on JAK2 and JAK3 is significantly weaker than that on JAK1, has high selectivity, and the selectivity ratio is significantly better than that of the control compounds. Specifically, the JAK1-JAK2 selectivity ratio of compounds 1A and 6 is 1.6 times and 2.9 times that of the control compound 1, respectively; the JAK1-JAK2 selectivity ratio of compounds 1A, 2A, 3, 4, 5 and 6 is 2.3-12.5 times that of the control compound 2A, and 3.3-19.3 times that of the control compound 2B; the JAK1-JAK3 selectivity ratio of compounds 1A, 2A, 3, 4, 5 and 6 is 8.8-50.3 times that of the control compound 2A, and 19.2-109.5 times that of the control compound 2B.

[0470] Table 7.4 Test results of the kinase activity (IC 50 , nM) of the compounds of the present application

[0471] Conclusion: The compounds of the present application have strong inhibitory activity on JAK1 and TYK2 kinase, and the inhibitory activity on JAK2 and JAK3 is significantly weaker than that on JAK1, with high selectivity.

[0472] Test Example 2: PBMC test

[0473] 1. Human peripheral blood mononuclear cell (PBMC) TYK2 inhibition activity test

[0474] Incubate human PBMC in 96-well plate at 37°C for 1 hour, then add different concentrations of test compound at 37°C for 1 hour; add IFN-a (final concentration of 30 ng / mL) and CD3 antibody (1.5 μL per well) at 37°C for 30 minutes; transfer to 96-well deep plate, add 1 mL of 1x Lyse / Fix buffer preheated at 37°C at 37°C for 10 minutes; centrifuge at 600g for 5 minutes, then wash twice with PBS, add Perm buffer III (400 μL per well) at 4°C for 30 minutes; centrifuge at 600g for 5 minutes, add 1 mL of Staining buffer (DPBS + 0.2% BSA + 1 mM EDTA) and centrifuge to wash twice; dilute Mouse anti-human Phospho-STAT5 (pY694) antibody 200-fold in Staining buffer, add 100 μL per well to the cell well, mix well, and incubate at room temperature for 40 minutes; add 1 mL / well of Staining buffer, centrifuge at 600g for 5 minutes and wash twice; after discarding the supernatant, resuspend the cell pellet in 200 μL of Staining buffer, and then load and analyze in a Beckman CytoFlex flow cytometer.

[0475] 2. Human peripheral blood mononuclear cell (PBMC) JAK1 inhibition activity test

[0476] Human PBMCs were seeded into 96-well plates and incubated at 37°C for 1 hour, then different concentrations of test compounds were added and incubated at 37°C for 1 hour; IL-6 (final concentration of 50 ng / mL) and CD3 antibody (1.5 μL per well) were added and incubated at 37°C for 30 minutes; transferred to 96-well deep plates, 1 mL of 1x Lyse / Fix buffer preheated at 37°C was added and incubated at 37°C for 10 minutes; centrifuged at 600g for 5 minutes, then washed twice with PBS, added Perm buffer III (400 μL per well) and incubated at 4°C for 30 minutes; centrifuged at 600g for 5 minutes, added 1 mL of Staining buffer (DPBS + 0.2% BSA + 1 mM EDTA) and centrifuged twice; Alexa Fluor 647 anti-STAT3 Phospho (Tyr705) Antibody was diluted 100-fold in Staining buffer, 100 μL per well was added to the cell wells, mixed well, and incubated at room temperature for 40 minutes; added 1 mL / well of Staining buffer, centrifuged at 600g for 5 minutes and washed twice; after discarding the supernatant, the cell pellets were resuspended in 200 μL of Staining buffer, and then loaded and analyzed in a Beckman CytoFlex flow cytometer.

[0477] 3. Human peripheral blood mononuclear cell (PBMC) JAK2 inhibitory activity test

[0478] Human PBMCs were seeded into 96-well plates and incubated at 37°C for 1 hour, then different concentrations of test compounds were added and incubated at 37°C for 1 hour; IL-6 (final concentration of 50 ng / mL) and CD3 antibody (1.5 μL per well) were added and incubated at 37°C for 30 minutes; transferred to 96-well deep plates, 1 mL of 1x Lyse / Fix buffer preheated at 37°C was added and incubated at 37°C for 10 minutes; centrifuged at 600g for 5 minutes, then washed twice with PBS, added Perm buffer III (400 μL per well) and incubated at 4°C for 30 minutes; centrifuged at 600g for 5 minutes, added 1 mL of Staining buffer (DPBS + 0.2% BSA + 1 mM EDTA) and centrifuged twice; Alexa Fluor 647 anti-STAT3 Phospho (Tyr705) Antibody was diluted 100-fold in Staining buffer, 100 μL per well was added to the cell wells, mixed well, and incubated at room temperature for 40 minutes; added 1 mL / well of Staining buffer, centrifuged at 600g for 5 minutes and washed twice; after discarding the supernatant, the cell pellets were resuspended in 200 μL of Staining buffer, and then loaded and analyzed in a Beckman CytoFlex flow cytometer.

[0479] The experimental results are shown in Table 8. Wherein, A represents IC 50 ≤ 10 nM, B represents 10 nM < IC 50 ≤ 100 nM, C represents 100 nM < IC 50 ≤ 500 nM, D represents IC 50 > 500 nM. The results show that the compounds 1A, 3 have strong inhibitory activity on human peripheral blood mononuclear cell (PBMC) TYK2, which is 5.6 times and 12.3 times higher than that of the control compound 1, respectively; the compounds 1A, 3 have strong inhibitory activity on human peripheral blood mononuclear cell (PBMC) JAK1, which is 4.5 times and 11.9 times higher than that of the control compound 1, respectively. The inhibitory activity of the compounds 1A, 3 on human peripheral blood mononuclear cell (PBMC) TYK2 and JAK1 is significantly better than that of the reference compound 1.

[0480] Table 8: PBMC test results

[0481] Conclusion: The compounds of the present application have strong inhibitory activity on JAK1 and TYK2, and weak inhibitory activity on JAK2, and have high selectivity.

[0482] Test Example 3: Effect of the Inflammatory Factor Secretion in the LPS-induced Microglial Cell Inflammation Model

[0483] The mouse microglial cell BV-2 cell line was recovered and cultured, and after the cell growth was observed to be 70-80% by microscopy, the test compound (compound concentration was 0.3 μM, 1.5 μM, 7.5 μM) was added, and after 1 hour, lipopolysaccharide (LPS) solution was added, and the final concentration of LPS was 100 ng / mL, an LPS-induced cell inflammation model was constructed, and after 24 hours, the cell supernatant was collected, and enzyme-linked immunosorbent assay (ELISA) was used to detect inflammatory factors interleukin 6 (IL-6), tumor necrosis factor-α (TNF-α), and chemokine 2 (CCL-2).

[0484] The experimental results are shown in Table 9. The results show that compared with the model group, after adding different concentrations of compound 3, the contents of inflammatory factors (IL-6, TNF-α, TNF-α) are significantly reduced, and show a dose-dependent manner; wherein, when the concentration of compound 3 is 1.5 μM and 7.5 μM, the inhibition rate of IL-6 is more than 100%.

[0485] Table 9: Effect of the compounds of the present application on the secretion of inflammatory factors

[0486] Conclusion: The compounds of the present application can significantly reduce the secretion of inflammatory factors (IL-6, TNF-α and CCL-2), and show a dose-dependent manner.

[0487] Test Example 4: Plasma protein binding experiment (equilibrium dialysis method)

[0488] The frozen CD-1 mice, Sprague-Dawley rats, beagle dogs, cynomolgus monkeys, and human plasma were thawed in flowing cold tap water. After the plasma was completely thawed, it was centrifuged at 3220 x g for 5 minutes and the supernatant and precipitate were removed. 597 μL of the blank plasma of each of the above species was taken, 3 μL of the working solution of the test sample or the control sample was added and mixed thoroughly to obtain a plasma sample (n = 1) with a test sample and a control sample concentration of 2 μM. The concentration of the organic phase DMSO was 0.5%. The sample was mixed thoroughly before the next step.

[0489] 50 μL of the plasma sample of the test sample and the control sample was taken into the sample receiving plate (n = 3), 50 μL of blank PBS was immediately added, and then 600 μL of the termination solution was added to the T0 sample of the test sample and the control sample, respectively, and stored at 2-8°C, waiting for subsequent processing with other dialyzed samples.

[0490] 100 μL of the plasma sample of the test sample and the control sample was added to the drug administration end of each dialysis hole (n = 3), and 100 μL of blank PBS was added to the receiving end corresponding to the dialysis hole. The dialysis plate was placed in a 5% CO2incubator and incubated at 37°C with about 100 rpm shaking for 4 hours.

[0491] After dialysis, 50 μL of the dialyzed PBS sample and the dialyzed plasma sample (n = 3) was taken into a new 96-well plate (sample receiving plate). The corresponding volume of the corresponding blank plasma or PBS was added to the sample so that the final volume of each sample well was 100 μL, and the volume ratio of plasma to PBS was 1:1. All samples were subjected to protein precipitation and then analyzed by LC-MS / MS.

[0492] The experimental results show that the free ratio of all test compounds in the plasma of each animal is greater than 30%, and the free ratio in human plasma is greater than 45%, without obvious species difference. A higher free drug ratio is beneficial to the in vivo efficacy.

[0493] Conclusion: The free drug concentration ratio of the compound of the present application in different species of plasma is high, and has good drug properties.

[0494] Test Example 5: Liver microsomal stability

[0495] The test substance was dissolved in DMSO to prepare a 10 mM DMSO solution, and then diluted with 100% acetonitrile to 100 μM to obtain a working solution (organic phase content: 99% acetonitrile, 1% DMSO).

[0496] Prepare two 96-well incubation plates, which are named as T60 incubation plate and NCF60 incubation plate, respectively.

[0497] Add 445 μL of microsomes working solution (the concentration of liver microsomal protein is 0.56 mg / mL) to the T60 incubation plate and NCF60 incubation plate, respectively, and then put the above-mentioned incubation plates in a 37°C water bath for pre-incubation for about 10 minutes.

[0498] After the pre-incubation, add 5 μL of test compound or control compound working solution to the T60 incubation plate and NCF60 incubation plate, respectively, and mix well.

[0499] Add 50 μL of potassium phosphate buffer to each well of the NCF60 incubation plate to start the reaction; add 180 μL of stop solution (containing 200 ng / mL tolbutamide and 200 ng / mL labetalol in acetonitrile) and 6 μL of NADPH regenerating system working solution to the T0 termination plate, and take 54 μL of sample from the T60 incubation plate to the T0 termination plate (T0 sample production). In the blank plate, only add 54 μL of microsomes working solution, 6 μL of NADPH regenerating system working solution and 180 μL of stop solution. Add 44 μL of NADPH regenerating system working solution to each well of the T60 incubation plate to start the reaction. Therefore, in the samples of test compound or control compound, the final concentrations of compound, testosterone, diclofenac and propafenone are 1 μM, the concentration of liver microsomes is 0.5 mg / mL, and the final concentrations of DMSO and acetonitrile in the reaction system are 0.01% (v / v) and 0.99% (v / v), respectively.

[0500] After incubation for an appropriate time (e.g. 5, 15, 30, 45 and 60 minutes), add 180 μL of stop solution (containing 200 ng / mL tolbutamide and 200 ng / mL labetalol in acetonitrile) to each well of the termination plate, and then take 60 μL of sample from the T60 incubation plate or NCF60 incubation plate to terminate the reaction.

[0501] Shake all sample plates and centrifuge at 3220 x g for 20 minutes, and then take 80 μL of supernatant per well and dilute in 240 μL of pure water for liquid chromatography tandem mass spectrometry analysis. The in vitro elimination rate constant ke of test compound and control compound is obtained by converting the ratio of peak area of compound to internal standard in the following formula into the percentage of remaining. The in vitro liver microsomal intrinsic clearance (CLint) is calculated by ke. int (mic) = 0.693 / T int (mic) = 0.693 / T 1 / 2 / microsomal protein content (the concentration of microsomes during incubation mg / mL). The experimental results are shown in Table 10.

[0502] Table 10 Test results of liver microsomal stability of compounds of the present application

[0503] Conclusion: The compounds of the present application have good stability in various species of liver microsomes.

[0504] Test Example 6: Hepatocyte stability

[0505] Several 96-well sample precipitation plates were prepared and named as TO, T15, T30, T60, T90, TO-MC, T90-MC and blank matrix, respectively. The recovery medium and incubation medium were taken out in advance and preheated in a 37°C water bath. The cryopreserved hepatocytes were taken out from the liquid nitrogen tank and immediately immersed in a 37°C water bath (about 90 seconds). After the cryopreserved part was loosened, it was poured into centrifuge tubes containing 40 mL of recovery medium, and the cells were resuspended in the recovery medium by gently inverting. At room temperature, 100 x g centrifugation for 5 minutes, remove the supernatant, resuspend the hepatocytes with an appropriate volume of incubation medium, and calculate the cell viability by trypan blue staining method. 198 μL of hepatocyte suspension (0.51 x 10 6 cells / mL) was added to the preheated incubation plate, and 198 μL of incubation medium without hepatocytes was added to the TO-MC and T120-MC incubation plates as the control group, and all the incubation plates were pre-incubated in a 37°C incubator for 10 minutes.

[0506] Then 2 μL of test sample and control compound working solution was added, mixed well, and immediately placed in the shaking plate machine in the incubator, and the timer was started to start the reaction. Two replicates were prepared for each time point of each compound. The incubation conditions were 37°C, saturated humidity, and 5% CO2.

[0507] In the test system, the final concentration of the test sample was 1 μM, the final concentration of the control sample was 3 μM, the final concentration of the hepatocytes was 0.5 x 10 6 cells / mL, and the final concentration of total organic solvent was 0.96%, of which the final concentration of DMSO was 0.1%. At the corresponding time point, the incubation was stopped, 25 μL of the mixture of compound and control compound and cells was taken out and added to a sample plate containing 125 μL of termination solution (200 ng / mL tolbutamide and labetalol in acetonitrile solution). For the blank sample plate, 25 μL of incubation medium without hepatocytes was added directly. After all the sample plates were sealed and shaken on the shaking plate machine at 600 rpm for 10 minutes, they were centrifuged at 3220 x g for 20 minutes. The supernatant of the test sample and control sample was diluted with ultrapure water at a ratio of 1:3. After all the samples were mixed, they were analyzed by LC / MS / MS method. The experimental results are shown in Table 11.

[0508] Table 11 Hepatocyte stability test results of the compounds of the present application

[0509] Conclusion: The compounds of the present application have good stability in various liver cells and exhibit moderate or slow metabolism.

[0510] Test Example 7: In vitro MDCKII-MDR1 monolayer cell permeability test

[0511] In this experiment, the MDR1-MDCK II cell line authorized by the Piet Borst laboratory of the Netherlands Cancer Institute was used as an in vitro model for permeability evaluation experiment, which is a Madin-Darby canine kidney cell (MDCK II) transfected with human multidrug resistance gene (MDR1), which can predict the permeability of compounds in barriers with high efflux effect such as duodenum, blood-brain barrier, liver cells and kidney units.

[0512] MDR1-MDCK II cells were seeded into Transwell-96-well cell plates, and after the cells formed a complete monolayer membrane structure, the transport experiment was carried out. The test concentration of the drug was 2.00 μM, and HBSS solution containing 10.0 mM HEPES (pH 7.40±0.05) was used as the transport buffer, and a bidirectional permeability study was carried out on the MDR1-MDCK II cell model, and the incubation was continued at 37°C, 5% CO2 for 150 minutes. The fluorescence yellow detection experiment was used to determine the integrity of the cell monolayer membrane, and low and high passive permeability controls and P-gp transport positive substrates were included in the experiment. After incubation, part of the samples from the donor chamber and the receiving chamber were transferred and extracted with acetonitrile containing appropriate internal standard (IS). The proteins were precipitated under the condition of 3220xg centrifugation for 20 minutes, and the supernatant was diluted with ultrapure water if necessary, and finally determined by liquid chromatography-tandem mass spectrometry (LC-MS / MS) technology using appropriate MRM transitions of analytes and internal standards. The apparent permeability coefficient (P app , cm / s) and efflux ratio (ER) were calculated according to the following equation.

[0513] wherein, V R is the volume of the receiving solution (0.075 mL for the A side and 0.25 mL for the B side); Area is the relative surface area of the cell monolayer (0.0804 cm 2 ); Time is the incubation time (9000 s); C0is the peak area ratio of the compound at the administration end; C R are the peak area ratios of the compounds at the administration end and the receiving end, respectively. The experimental results show that the solvent recovery rates of the tested compounds of the present application are all greater than 90%, and the specific P app and ER data are shown in Table 12.

[0514] Table 12. Results of MDCKII-MDR1 cell permeability test of compounds of the present application

[0515] Conclusion: The compounds of the present application have high recovery rate in MDCKII-MDR1 monolayer cell permeability experiment, showing high permeability and low efflux.

[0516] Test Example 8: Cytochrome P450 enzyme (CYP) inhibition study

[0517] 1. Test purpose: To determine the inhibitory effect of the test compound on the activity of human liver microsomal cytochrome P450 isozymes (CYP1A2, CYP2C9, CYP2C19, CYP2D6 and CYP3A4).

[0518] 2. Experimental method

[0519] The test compound (10.0 mM) was gradiently diluted to prepare a working solution (100x final concentration) with concentrations of 5.00, 1.50, 0.500, 0.150, 0.0500, 0.0150 and 0.00500 mM, respectively. Meanwhile, working solutions of positive inhibitors of P450 isozymes (CYP1A2, CYP2C9, CYP2C19, CYP2D6 and CYP3A (with midazolam as a probe substrate)) and their specific substrate mixtures were prepared. Human liver microsomes stored in a refrigerator below -60°C were thawed on ice, and then diluted with potassium phosphate buffer (PB) to prepare a working solution with a certain concentration (0.253 mg / mL).

[0520] 20.0 μL of the substrate mixture was added to the reaction plate (20.0 μL of PB was added to the blank wells), and then 158 μL of the human liver microsomal working solution was added to the reaction plate, which was placed on ice for use. At this time, 2.00 μL of each concentration of the test compound (N=1) and the specific inhibitor (N=2) was added to the corresponding wells, and the non-inhibitor (without the test compound or the positive inhibitor) group was added with the corresponding organic solvent as the control group sample (the test compound control sample was DMSO:MeOH=1:1, and the positive control sample was DMSO:MeOH=1:9). After pre-incubation at 37°C for 10 min, 20.0 μL of coenzyme factor (NADPH) solution was added to the reaction plate, which was incubated at 37°C for 10 min. The reaction was terminated by adding 400 μL of pre-cooled acetonitrile solution (containing internal standard). The reaction plate was placed on a shaker and shaken for 10 min to mix. Then it was centrifuged at 4°C and 4000 rpm for 20 min. 200 μL of supernatant was added to 100 μL of water for sample dilution. Finally, the plate was sealed and shaken for 10 min to mix, and then subjected to LC-MS / MS detection.

[0521] The experimental results show that the inhibition IC of compounds 1A, 2A and 3 on human liver microsomal cytochrome P450 isozymes (CYP1A2, CYP2C9, CYP2C19, CYP2D6 and CYP3A4) is greater than 30 μM. 50

[0522] Conclusion: The compounds of the present application have no obvious inhibition on the five main cytochrome P450 enzymes of human liver microsomes, and the risk of “drug-drug interaction” is low.

[0523] Test Example 9: PXR / CAR / AhR induction study

[0524] The PXR experiment uses HepG2-human PXR+CYP3A4 stable cell strain; the CAR experiment uses HepG2 cell strain; and the AhR test uses HepG2-Lucia TM AhR cells stable cell strain.

[0525] First, the test compound (30.0 mM) is gradiently diluted to prepare a working solution (100x final concentration), and the working solution concentrations are 3.00, 1.00, 0.333, 0.111, 0.0370, 0.0123, 0.00412, 0.00137 and 0.000457 mM, respectively.

[0526] CAR induction experiment method:

[0527] The CAR test uses HepG2 cell strain, and the cells are subcultured after resuscitation. When the cell growth density reaches 90%, the cells are counted and digested, 3*10 5 cells / mL, 20 wells for each compound (n=2), 100 μL / well, and plated in a 96-well plate. Incubate in a 37°C incubator for 24 hours. Transfect CYP2B6 and CAR into the cells, shake well, and incubate in an incubator overnight. Discard the culture medium in the cell plate, and add 90 μL of culture medium again. Take 3 μL of working solution, dilute 30 times in 87 μL of EMEM culture medium, mix well, and add 10 μL of the drug to be added to the cell plate, so that the high dose = 100 μM, DMSO as negative control group, and the DMSO content is 0.3%, and incubate at 37°C for 24 hours. Take the cell culture plate out of the incubator, and place it at room temperature for 30 min to balance the temperature of the culture plate to room temperature. Equilibrate the Bright-Lite Luciferase Assay System reagent to room temperature, add 50 μL per well, and shake at 200 rpm for 3 min. Read the Luciferase signal value on the enzyme marker (BMG). Use GraphPad Prism 8 to draw the EC​50 Graph.

[0528] PXR induction experiment method:

[0529] PXR experiment adopts HepG2-human PXR+CYP3A4 stable transfection cell strain; when the cell growth density reaches 90%, the cell is counted and digested, 3*10 5 cells / mL, 20 holes of each compound (n=2), 100 μL / well, and is placed in a 96-hole plate; it is incubated in a 37°C incubator for 24 hours. The culture medium in the cell plate is discarded, and 90 μL of culture medium is added again. 3 μL of working solution is taken and diluted 30 times in 87 μL of EMEM culture medium, and mixed, as a drug to be added; 10 μL of the drug to be added is taken and added to the cell plate, so that the high dose = 100 μM, DMSO is used as a negative control group, and the DMSO content is 0.3%, and is incubated at 37°C for 24 hours. The cell culture plate to be tested is taken out of the incubator, and is placed at room temperature for 30 min, so that the temperature of the culture plate is balanced to room temperature, the Bright-Lite Luciferase Assay System reagent is balanced to room temperature, 50 μL is added to each hole, and is shaken at 200 rpm for 3 min, and the Luciferase signal value is read on an enzyme marker (BMG). The EC 50 Graph.

[0530] AhR induction experiment method:

[0531] AhR test adopts resuscitation HepG2-Lucia TM AhR cells stable transfection cell strain, and is subcultured; when the cell growth density reaches 90%, the cell is counted and digested, 4000 cells / well / 40 μL, 20 holes of each compound (n=2), and is placed in a 384-hole plate; it is incubated in a 37°C incubator for 24 hours. The culture medium in the cell plate is discarded, and 39 μL of culture medium is added again. Compound preparation: 4 μL of working solution is taken and diluted 7.5 times in 26 μL of EMEM culture medium, and mixed, as a drug to be added; 1 μL of the drug to be added is taken and added to the cell plate, so that the high dose = 100 μM, DMSO is used as a negative control group, and the DMSO content is 0.3%, and is incubated at 37°C for 24 hours. The cell culture plate to be tested is taken out of the incubator, and is placed at room temperature for 30 min, so that the temperature of the culture plate is balanced to room temperature, the QUANTI-Luc TM Gold reagent is balanced to room temperature, 4 μL of cell culture supernatant is taken into a 384-hole plate, 20 μL of QUANTI-Luc TMGold, 1000 rpm centrifugation for 1 min, and read the Luciferase signal value on the microplate reader (BMG). 5) EC 50 graphs.

[0532] The experimental results show that the EC 50 of compound 1A and compound 3 are both greater than 30 μM at the highest experimental concentration (30 μM).

[0533] Conclusion: The compound of the present application has no obvious induction effect on CAR, PXR and AhR, which indicates that the compound of the present application has very low induction risk on major cytochrome P450 enzymes.

[0534] Test Example 10: hERG test

[0535] CHO cells stably expressing hERG were cultured in 35 mm diameter cell culture dishes in a 37°C, 5% CO2 incubator and passaged at a 1:5 ratio every 48 hours. On the day of the experiment, the cell culture medium was aspirated and the cells were rinsed once with extracellular fluid before the addition of 0.25% Trypsin-EDTA (Invitrogen) solution for 3-5 minutes at room temperature. The trypsin solution was aspirated and the cells were resuspended in extracellular fluid before being transferred to experimental dishes for electrophysiological recording.

[0536] The test compound was prepared as a 20 mM stock solution in DMSO, and then serially diluted 3-fold with DMSO, i.e. 10 μL was added to 20 μL of DMSO, and then 10 μL of each serially diluted compound DMSO solution was added to 4990 μL of extracellular fluid to obtain the final concentration required for testing after 500-fold dilution.

[0537] CHO cells stably expressing hERG potassium channels were used to record hERG potassium channel currents using the whole-cell voltage clamp technique at room temperature. After obtaining the whole-cell recording, the cells were clamped at -100 mV, and the step voltage for inducing hERG potassium current (I hERG) was given a 2s depolarization voltage from -100 mV to +20 mV, and then repolarized to -50 mV for 1s before returning to -100 mV. This voltage stimulation was given every 5s, and after the hERG potassium current was determined to be stable (1 minute), the drug administration process was started. Each test concentration of the compound was given for at least 1 minute to reach a steady state or for a maximum of 3 minutes, and at least 2 cells were tested for each concentration (n≥2).

[0538] Data analysis was processed by pClamp and Excel software. The inhibition degree of different compound concentrations on hERG potassium current (hERG tail current peak induced at -50 mV) was calculated by the following formula: Inhibition% = [1 - (I / I0)] x 100%. Wherein, Inhibition% represents the inhibition rate of the compound on hERG potassium current, and I and I0 represent the amplitude of hERG potassium current after and before adding the drug, respectively. GraphPad Prism 8 was used for plotting and calculating IC 50 .

[0539] The experimental results show that the hERG IC 50 >40 μM.

[0540] Conclusion: The compound of the present application has no obvious inhibition on hERG.

[0541] Test Example 11: PK study

[0542] 1. 6 male C57 / 6J mice were divided into 2 groups, 3 animals in each group. The intravenous (iv) group was administered at a dose of 2 mg / kg, and the solvent was 5% DMSO / 10% Solutol / 85% Saline; the oral administration (po) group was administered at a dose of 15 mg / kg, and the solvent was 0.5% HPMC / water.

[0543] 2. 6 male SD rats were divided into 2 groups, 3 animals in each group. The intravenous (iv) group was administered at a dose of 2 mg / kg, and the solvent was 5% DMSO / 10% Solutol / 85% Saline; the oral administration (po) group was administered at a dose of 10 mg / kg, and the solvent was 0.5% HPMC / water (the po group solvent of compound 1A was 0.5% HPMC / 0.2% tween80 / water).

[0544] 3. 6 male beagle dogs were divided into 2 groups, 3 animals in each group. The intravenous (iv) group was administered at a dose of 0.5 mg / kg, and the solvent was 5% DMSO / 20% PEG400 / water; the oral administration (po) group was administered at a dose of 2 mg / kg, and the solvent was 0.5% HPMC / 0.2% tween80 / water.

[0545] Whole blood was collected at 5 min (only iv group), 15 min, 30 min, 1 h, 2 h, 4 h, 8 h and 24 h after administration, respectively, and the whole blood was placed in an anticoagulant tube containing EDTA-K2, and centrifuged to prepare plasma. The concentration of the test molecule in the plasma was quantitatively detected by LC-MS / MS, and the PK parameters were calculated by Phoenix WinNonlin.

[0546] The experimental results are shown in Table 13. Note: CL represents clearance, Vdss represents distribution volume, T 1 / 2 is half-life, AUC 0-last represents the area under the whole blood concentration-time curve from 0 to the last quantifiable time point, C max is peak concentration; F represents bioavailability.

[0547] Table 13. PK test results of the compound of the present application

[0548] Conclusion: The compound of the present application exhibits high oral exposure and high oral bioavailability in mice, rats and dogs, with small species difference, and has excellent pharmacokinetic properties.

[0549] Test Example 12: Brain entry test

[0550] 1. 12 male C57 / 6J mice were divided into 4 groups, 3 animals in each group. All animals were orally administered with 15 mg / kg, and the solvent was 0.5% HPMC aqueous solution. At 0.5, 2, 4 and 8 hours after administration, 1 group of animals was sacrificed respectively, and whole blood and whole brain (brain) were collected. The whole blood was placed in an anticoagulant tube containing EDTA-K2, and plasma was prepared by centrifugation. The brain tissue was homogenized with PBS. The concentration of the test molecule in plasma and brain homogenate (ng / mL in plasma, ng / g in brain homogenate) was quantitatively detected by LC-MS / MS, and the PK parameters were calculated by Phoenix WinNonlin.

[0551] 2. 9 male SD rats were divided into 3 groups, 3 animals in each group. All animals were orally administered with 10 mg / kg, and the solvent was 0.5% HPMC / 0.2% tween80 / water. At 0.5, 2 and 8 hours after administration, 1 group of animals was sacrificed respectively, and whole blood and whole brain (brain) were collected. The whole blood was placed in an anticoagulant tube containing EDTA-K2, and plasma was prepared by centrifugation. The brain tissue was homogenized with PBS. The concentration of the test molecule in plasma and brain homogenate was quantitatively detected by LC-MS / MS, and the PK parameters were calculated by Phoenix WinNonlin. The experimental results are shown in Table 14.

[0552] Table 14. Brain entry test results of the compound of the present application

[0553] Conclusion: The compound of the present application has good brain tissue distribution in mice and rats.

[0554] Test Example 13: In vivo efficacy (I)

[0555] Multiple sclerosis (MS) is a chronic inflammatory neurodegenerative disease of the central nervous system. Experimental autoimmune encephalomyelitis (EAE) induced by myelin oligodendrocyte glycoprotein (MOG) is the most common animal model of human multiple sclerosis. The present experiment aims to test the in vivo efficacy of the compounds of the present application in the EAE model.

[0556] Experimental methods:

[0557] 1. Model establishment:

[0558] After the adaptation period, C57BL / 6J mice (female, about 7 weeks old) were subcutaneously injected with 200 μL MOG emulsion (referred to as Day 1), in which the final concentration of MOG was 1.5 mg / mL and the concentration of Mycobacterium tuberculosis / complete Freund's adjuvant (CFA) solution was 4 mg / mL. On the day of injection and the next day, the mice were intraperitoneally injected with 250 μL / each of pertussis toxin (PTX, 1.0 μg / mL). During the experiment, the body weight, disease occurrence and clinical score of the mice were observed every day. The scoring criteria were as follows: 0, normal, no obvious symptoms; 1, tail weakness or slight hind limb weakness; 2, tail weakness and hind limb weakness, which can be recovered by passive turning over; 3, unilateral hind limb hemiplegia, which cannot be recovered by passive turning over; 4, complete hind limb paralysis, forelimb paralysis or muscle weakness, accompanied by urinary and fecal incontinence; 5, moribund state or death.

[0559] 2. Drug administration:

[0560] Oral gavage was started on Day 13 and administered twice a day (BID) until Day 24. The dosages were as follows: Compound 1A: 10 mg / kg and 30 mg / kg; Compound 3: 3 mg / kg, 10 mg / kg and 20 mg / kg. The solvent was 0.5% HPMC / water.

[0561] 3. Experimental results:

[0562] The experimental results of the clinical score of the animals in the EAE model are shown in Figure 1.1, and the experimental results of the body weight change of the animals are shown in Figure 1.2. Specifically, after administration of Compound 1A and Compound 3, the clinical score was significantly reduced in a dose-dependent manner, and in the experimental groups of Compound 1A (30 mg / kg) and Compound 3 (20 mg / kg), no obvious symptoms were observed in the animals from Day 13 to Day 24, and the clinical score remained at 0. In terms of body weight, the animals in the Compound 1A and Compound 3 administration groups recovered faster than the animals in the model group on the 22nd day of administration. After administration of the compounds, the body weight was significantly improved in a dose-dependent manner.

[0563] Conclusion: The compounds of the present application exhibit significant therapeutic effects in the EAE efficacy model, and show a positive dose-effect correlation.

[0564] Test Example 14: In vivo efficacy (II)

[0565] Purpose of experiment: test the in vivo efficacy of the compounds of the present application in EAE model.

[0566] Experimental method:

[0567] 1. Model establishment:

[0568] C57BL / 6J mice (female, about 7 weeks old) were subcutaneously injected with 200 μL MOG emulsion (referred to as Day 1) after the adaptation period, the final concentration of MOG in the emulsion was 1.5 mg / mL, and the concentration of Mycobacterium tuberculosis / complete Freund's adjuvant (CFA) solution was 4 mg / mL. On the day of injection of MOG and on the third day, pertussis toxin (PTX, 1.0 μg / mL) was injected intraperitoneally, the dose was 250 μL per mouse. Body weight, disease occurrence and clinical score were observed every day during the experiment. The scoring criteria were the same as those in Test Example 13.

[0569] 2. Drug administration:

[0570] Oral gavage was started on Day 16 and continued twice a day (BID) until Day 30. The dosages were as follows: reference compound 1: 30 mg / kg; compound 3: 1.5 mg / kg, 3 mg / kg, 7.5 mg / kg and 15 mg / kg. The solvent was 0.5% HPMC / water.

[0571] 3. Experimental results:

[0572] The experimental results of the clinical scores of the animals in the EAE model are shown in Figure 2, and the results of the area under the curve from Day 17 to Day 30 are shown in Table 15. Specifically, after administration of the reference compound 1 and compound 3, the clinical scores were significantly reduced, and compound 3 showed a dose-dependent effect. The clinical scores and the area under the curve of the experimental groups administered with compound 3 (7.5 mg / kg, 15 mg / kg) were lower than those of the group administered with the reference compound 1 (30 mg / kg), indicating that compound 3 can achieve the same efficacy as the reference compound 1 at a lower dose.

[0573] Table 15 Area under the curve results

[0574] Conclusion: The compounds of the present application exhibit excellent therapeutic effect in the EAE efficacy model, have low effective dose, and show a positive dose-effect correlation.

Claims

1. A compound of Formula (II), a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, wherein, T1, T2, and T3 are independently selected from N and CR3; ring A is selected from ring A1 and ring A2; Ring A1 is arbitrarily selected by one or more R a1 Substituted monocyclic 4-8 membered oxoheterocyclic alkyl groups; Ring A2 is wherein 1 represents the connection site with the imidazole N, and 2 represents the connection site with R4; Ring B is selected from one or more R. b Substituted monocyclic 4-8 membered nitrogen-containing heterocyclic alkyl groups; ring C is fused to ring B and ring C is selected from optionally substituted 5-10 membered nitrogen-containing heteroaryl; c substituted 5-10 membered nitrogen-containing heteroaryl; T is N, which is optionally quaternized or oxidized; when ring A is ring A1, T1 is CR3, T2 is CR3, and T3 is O or NH; when ring A is ring A2, one of T1, T2, and T3 is O, and the other two are independently selected from N and CR3; R1is -L1-R 11 ; L1is selected from the group consisting of a bond, -CONR 12 , -CO-, O, S, NR 12 , and the following groups optionally substituted with 1 or more R 1a : C 1-4 1-6alkyl, C 2- 2-6alkenyl, C 2-4 2-6alkynyl, C 1-4 1-6alkoxy, C 1-4 1-6alkylthio, C 1-4 1-6alkylamino, C 3-8 cycloalkyl, 3-8 membered heterocycloalkyl, phenyl, and 5-6 membered heteroaryl; R 11 selected from H, D, F, CI, Br, I, CN, and optionally substituted by 1 or more R 1a substituted by 1 or more R 1-4 substituted by 1 or more R 2-4 substituted by 1 or more R 2-4 substituted by 1 or more R 1-4 substituted by 1 or more R 1-4 substituted by 1 or more R 1-4 substituted by 1 or more R 3-8 substituted by 1 or more R R 12 is selected from H and optionally substituted with 1 or more R 1a is selected from H, C 1-4 alkyl, C 2-4 alkenyl, C 2-4 alkynyl, C 3-8 cycloalkyl and 3-8 membered heterocycloalkyl; R2is selected from the group consisting of H, D, F, CI, Br, I, OH, NH2, CN, and optionally substituted with 1 or more R 2a substituted C 1-4 alkyl, C 2-4 alkenyl, C 2-4 alkynyl, C 1-4 alkoxy, C 1-4 alkylthio, C 1-4 alkylamino, C 3-8 cycloalkyl, 3-8 membered heterocycloalkyl, phenyl, and 5-6 membered heteroaryl; R3 is selected from H, D, F, Cl, Br, I, OH, NH2, CN, and optionally one or more R 3a The following groups are substituted: C 1-4 Alkyl, C 2-4 alkenyl, C 2-4 alkynyl group, C 1-4 Alkoxy, C 1-4 Alkylthio, C 1-4 Alkylamino, C 3-8 Cycloalkyl, 3-8 membered heterocycloalkyl, phenyl, and 5-6 membered heteroaryl; or, two R3s on adjacent atoms are linked to form an array optionally bounded by one or more R3s. 3b The following groups are substituted: C 5-8 Cycloalkyl, 5-8 membered heterocycloalkyl or 5-6 membered heteroaryl; R4is selected from CN, C 1-4 alkyl, C 2-4 alkenyl, C 2-4 alkynyl, C 1-4 alkoxy, C 1-4 alkylthio, C 1-4 alkylamino, C 3- 8cycloalkyl, 3-8 membered heterocycloalkyl, phenyl and 5-6 membered heteroaryl, which C 1-4 alkyl, C 2-4 alkenyl, C 2-4 alkynyl, C 1-4 alkoxy, C 1- alkylthio, C 3-8 cycloalkyl, 3-8 membered heterocycloalkyl, phenyl and 5-6 membered heteroaryl is substituted with CN, and furthermore optionally substituted with one or more R 4a substituents; each R a1 , each R b , each R c , each R 1a , R 11a , R 11b , R 11c , R 11d , each R 2a , each R 3a , each R 3b , each R 4a are each independently selected from the group consisting of H, D, F, Cl, Br, I, OH, NH2, CN, =O, and the following groups optionally substituted with 1 or more R: C 1-4 alkyl, C 2-4 alkenyl, C 2-4 alkynyl, C 1-4 alkoxy, C 1-4 alkylthio, C 3-8 cycloalkyl, 3-8 membered heterocycloalkyl, phenyl, and 5-6 membered heteroaryl; or 2 R a1 or 2 R b or 2 R c or 2 R b and R c together, each independently form an optionally substituted C 3-8 cycloalkyl, 4-8 membered heterocycloalkyl, phenyl or 5-6 membered heteroaryl; each R is independently selected from H, D, F, CI, Br, I, OH, NH2, CN, and the following groups optionally substituted with 1 or more F: C 1-4 alkyl, C 2-4 alkenyl, C 2-4 alkynyl, C 1-4 alkoxy, C 1-4 alkylthio, C 1-4 alkylamino, C 3-8 cycloalkyl, 3-8 membered heterocycloalkyl, phenyl, and 5-6 membered heteroaryl.

2. The compound, stereoisomer, or pharmaceutically acceptable salt thereof according to claim 1, wherein, each R is independently selected from H, D, F, Cl, OH, NH2, CN, CH3, and CF3.

3. The compound, stereoisomer, or pharmaceutically acceptable salt thereof according to claim 1, wherein, each R a1 are each independently selected from the group consisting of H, D, F, CI, Br, I, OH, NH2, CN, and optionally substituted with 1 or more R, CH3, CH2CH3, vinyl, ethynyl, OCH3, OCH2CH3, cyclopropyl, and cyclobutyl; further, each R a1 are each independently selected from the group consisting of H, D, F, CI, CH3, and CF3.

4. The compound, stereoisomer, or pharmaceutically acceptable salt thereof according to claim 1, wherein, each R b , each R c is independently selected from the group consisting of H, D, F, Cl, Br, I, OH, NH2, CN, =0, and the following groups optionally substituted with 1 or more R: CH3, CH2CH3, CH2CH2CH3, CH(CH3)2, vinyl, propenyl, ethynyl, propynyl, OCH3, OCH2CH3, OCH(CH3)2, SCH3, SCH2CH3, cyclopropyl, and cyclobutyl; further, each R b , each R c is independently selected from the group consisting of H, F, Cl, CH3, and CF3.

5. The compound, stereoisomer, or pharmaceutically acceptable salt thereof according to claim 1, wherein, each R 1a , R 11a , R 11b , R 11c , R 11d , each R 2a , each R 3a , each R 3b , each R 4a is independently selected from the group consisting of H, D, F, Cl, Br, I, OH, NH2, CN, =0, and the following groups optionally substituted with 1 or more R: CH3, CH2CH3, CH2CH2CH3, CH(CH3)2, vinyl, propenyl, ethynyl, propynyl, OCH3, OCH2CH3, OCH(CH3)2, SCH3, SCH2CH3, cyclopropyl, and cyclobutyl; further, each R 1a , R 11a , R 11b , R 11c , R 11d , each R 2a , each R 3a , each R 3b , each R 4a is independently selected from the group consisting of H, D, F, Cl, CN, =0, CH3, CFH2, CF2H, CF3, OCH3, OCF3, and cyclopropyl.

6. The compound, stereoisomer, or pharmaceutically acceptable salt thereof of claim 1, wherein, L1is selected from the group consisting of a bond, C(O), O, S, and the following groups optionally substituted with 1 or more F or D: CH2, CH2CH2, OCH2, OCH2CH2, and cyclopropyl, R 1a substituted with 1 or more F or D: CH3, CH2CH3, CH2CH2CH3, CH(CH3)2, OCH3, OCH2CH3, OCH(CH3)2, cyclopropyl, cyclobutyl, oxetanyl, oxetanyl, 11 substituted with 1 or more F or D: CH3, CH2CH3, CH2CH2CH3, CH(CH3)2, OCH3, OCH2CH3, OCH(CH3)2, cyclopropyl, cyclobutyl, oxetanyl, oxetanyl, substituted with 1 or more F or D: CH3, CH2CH3, CH2CH2CH3, CH(CH3)2, OCH3, OCH2CH3, OCH(CH3)2, cyclopropyl, cyclobutyl, oxetanyl, oxetanyl, 1a substituted with 1 or more F or D: CH3, CH2CH3, CH2CH2CH3, CH(CH3)2, OCH3, OCH2CH3, OCH(CH3)2, cyclopropyl, cyclobutyl, oxetanyl, oxetanyl, 11 substituted with 1 or more F or D: CH3, CH2CH3, CH2CH2CH3, CH(CH3)2, OCH3, OCH2CH3, OCH(CH3)2, cyclopropyl, cyclobutyl, oxetanyl, oxetanyl, 7. The compound, stereoisomer, or pharmaceutically acceptable salt thereof of claim 1, wherein, R1 is selected from H, D, F, Cl, Br, I, CN, and optionally one or more R 1a The following groups are substituted: C 1-4 Alkyl, -OC 1-4 Alkyl and -SC 1-4 Alkyl; further, R1 is selected from H, D, F, Cl, Br, I, CN, CH3, CFH2, CF2H, CF3, CD3, CH2CH3, CH(CH3)2, CH2CN, CH2CH2CN, OCH3, OCF3, OCD3, OCH2CH3, OCH(CH3)2, SCH3, SCF3, SCH2CH3.

8. The compound, stereoisomer, or pharmaceutically acceptable salt thereof of claim 1, wherein, R2 is H, and R3 is H.

9. The compound, stereoisomer, or pharmaceutically acceptable salt thereof of claim 1, wherein, R4is selected from the group consisting of CN, CH3, CH2CH3, CH2CH2CH3, CH(CH3)2, vinyl, propenyl, ethynyl, propynyl, OCH3, OCH2CH3, OCH(CH3)2, SCH3, SCH2CH3, cyclopropyl and cyclobutyl, said CH3, CH2CH3, CH2CH2CH3, CH(CH3)2, vinyl, propenyl, ethynyl, propynyl, OCH3, OCH2CH3, OCH(CH3)2, SCH3, SCH2CH3, cyclopropyl and cyclobutyl being further optionally substituted with 1 or more R 4a substituents; further, R4is selected from the group consisting of CH2CN.

10. The compound, stereoisomer, or pharmaceutically acceptable salt thereof, of claim 1, wherein, Ring A1 is selected from one or more Rs. a1 Substituted oxetyl, oxetyl, oxetylhexyl, oxetylheptyl, and oxetylhexyl; further, ring A1 is selected from...

11. The compound, stereoisomer, or pharmaceutically acceptable salt thereof, of claim 1, wherein, Ring A2is selected from 12. The compound, stereoisomer, or pharmaceutically acceptable salt thereof, of claim 1, wherein, structural units selected from further, structural units selected from 13. The compound, stereoisomer, or pharmaceutically acceptable salt thereof, according to any one of claims 1 to 12, which is selected from: wherein, m and n are independently selected from 0, 1, 2, and 3; R1is -L1-R 11 ; L1is selected from the group consisting of a bond, -CONH-, -CO-, O, S, NH, and optionally substituted with 1 or more R 1a substituted C 1-4 alkyl, C 2-4 alkenyl, C 2-4 alkynyl, C 1-4 alkoxy, C 1-4 alkylthio, C 1-4 alkylamino, C 3-8 cycloalkyl and 3-8 membered heterocycloalkyl; R 11 H, F, CI, Br, I, CN and optionally substituted by one or more R 1a C 1-4 alkyl, C 2-4 alkenyl, C 2- 4alkynyl, C 1-4 alkoxy, -C 3-8 cycloalkyl and 3-8 membered heterocycloalkyl; R2is selected from H, F, CI, Br, I, OH, NH2, CN, and optionally substituted with 1 or more R 2a C1-8alkyl, C 1-4 C1-8alkyl, C 2-4 C1-8alkyl, C 2-4 C1-8alkyl, C 1-4 C1-8alkyl, C 3-8 C1-8alkyl, C R3is selected from H, F, CI, Br, I, OH, NH2, CN, and optionally substituted with 1 or more R 3a C1-8alkyl, C 1-4 C1-8alkyl, C 2-4 C1-8alkyl, C 2-4 C1-8alkyl, C 1-4 C1-8alkyl, C 3-8 C1-8alkyl, C R4is selected from CN, CH3, CH2CH3, CH2CH2CH3, CH(CH3)2, vinyl, propenyl, ethynyl, propynyl, OCH3, OCH2CH3, OCH(CH3)2, SCH3, SCH2CH3, cyclopropyl, and cyclobutyl, said CH3, CH2CH3, CH2CH2CH3, CH(CH3)2, vinyl, propenyl, ethynyl, propynyl, OCH3, OCH2CH3, OCH(CH3)2, SCH3, SCH2CH3, cyclopropyl, and cyclobutyl being further optionally substituted with 1 or more R 4a substituents; Ring B is selected from one or more R. b Substituted monocyclic 5-7 member nitrogen-containing heterocyclic alkyl groups; ring C is fused to ring B and ring C is selected from optionally substituted 5-6 membered nitrogen containing heteroaryl; c substituted 5-6 membered nitrogen containing heteroaryl; R a1 , R 2a , R 3a , R 4a , R b , R c are each independently selected from the group consisting of H, F, Cl, Br, I, OH, NH2, CN, =0 and the following groups optionally substituted with 1 or more F: C 1-4 alkyl, C 2-4 alkenyl, C 2-4 alkynyl, C 1-4 alkoxy, C 1-4 alkylthio, C 3-6 cycloalkyl, 3-6 membered heterocycloalkyl.

14. The compound of formula (I-1), a stereoisomer thereof, or a pharmaceutically acceptable salt thereof according to claim 13, wherein, m is selected from 0, 1, 2, and 3; n is selected from 1, 2, and 3; R1is selected from F, CI, Br, I, CN, and optionally substituted with one or more R 1a substituted C 1-4 alkyl, -O-C 1-4 alkyl and -S-C 1-4 alkyl; R2 is selected from H, F, Cl, Br, I, and CH3; each R3 is independently selected from H, F, Cl, Br, I, and CH3; R4is selected from CN, CH2CN, CH2CH2CN, CH(CH3)CN, C(CH3)2CN, Each R 1a They were each independently selected from H, D, F, Cl, and CN; each R is independently selected from the group consisting of H, F, Cl, Br, I, CN, =0, and C1-6alkyl optionally substituted with 1 or more F; a1 each R is independently selected from the group consisting of H, F, Cl, Br, I, CN, =0, and C1-6alkyl optionally substituted with 1 or more F; 1-4 alkyl.

15. The compound, stereoisomer, or pharmaceutically acceptable salt thereof, of claim 13 or 14 selected from: wherein, m and s are independently selected from 0, 1, 2, and 3; R1, R2, R3, R a1 , R b , R c as defined in claim 13 or 14.

16. The compound, stereoisomer, or pharmaceutically acceptable salt thereof, of claim 15 is selected from: wherein, R1is selected from F, CI, Br, I, CN, and optionally substituted with one or more R 1a substituted C 1-4 alkyl, -O-C 1-4 alkyl and -S-C 1-4 alkyl; Each R 1a They were each independently selected from H, D, F, Cl, and CN; R4is selected from CN, CH2CN, CH2CH2CN, CH(CH3)CN, C(CH3)2CN, 17. The compound, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof according to claim 16, wherein R1 is selected from F, Cl, Br, I, CN, and CH3 or OCH3 optionally substituted with 1, 2, or 3 F or D.

18. The compound, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof according to claim 17, wherein R4 is selected from CH2CN.

19. A compound represented by Table A and / or Table A1, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof.

20. A crystalline form of Compound 3, having characteristic X-ray powder diffraction peaks at the following 2θ angles: 8.45±0.20°, 9.98±0.20°, and 13.60±0.20°.

21. The B crystal form according to claim 20, having an X-ray powder diffraction pattern, with Cu Kα radiation, comprising at least 5, 6, 7, or 8 diffraction peaks selected from the following 2θ angles: 8.45±0.20°, 9.98±0.20°, 13.60±0.20°, 18.13±0.20°, 19.22±0.20°, 21.94±0.20°, 23.91±0.20°, and 24.77±0.20°.

22. The B crystal form according to claim 21, having an X-ray powder diffraction pattern, with Cu Kα radiation, having characteristic X-ray powder diffraction peaks at the following 2θ angles: 8.45°, 9.98°, 11.88°, 13.60°, 16.91°, 18.13°, 19.22°, 20.02°, 21.26°, 21.76°, 21.94°, 22.15°, 23.02°, 23.91°, 24.25°, 24.77°, 25.43°, 26.01°, 27.36°, 28.89°, 29.61°, and 30.50°.

23. The B crystal form according to any one of claims 20 to 22, further having any one of the following characteristics: (1) an XRPD pattern substantially as shown in Figure 6; (2) a differential scanning calorimetry curve having an onset value for the endothermic peak at 249.5±3°C; (3) a DSC pattern substantially as shown in Figure 7; (4) a thermogravimetric analysis curve having a weight loss of 0.25% in the range of 30.0±3°C to 150.0±3°C; (5) a TGA pattern substantially as shown in Figure 8.

24. Use of the compound of any one of claims 1-19, a stereoisomer thereof or a pharmaceutically acceptable salt thereof, or the B crystal form of any one of claims 20-23 in the manufacture of a medicament for the treatment of a JAK1 / TYK2 inhibitor-related disease.

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