Substituted tricyclic derivatives and uses thereof

Abstract

The invention provides a series of substituted tricyclic derivatives and application thereof, and particularly provides a compound as shown in formula (II), (I-1) or (I-2), and a stereoisomer and pharmaceutically acceptable salt thereof.

Description

[0001] Cross-references to related applications

[0002] This application claims priority to two earlier patent applications filed with the China National Intellectual Property Administration on August 8, 2024 (application number CN2024110886648) and January 24, 2025 (application number CN2025101225477), the entire contents of which are incorporated herein by reference. Technical Field

[0003] This invention relates to a series of substituted tricyclic derivatives and their applications, particularly to compounds of formulas (II), (I-1), and (I-2), their stereoisomers, and pharmaceutically acceptable salts thereof. Background Technology

[0004] Janus kinases (JAKs) are cytoplasmic tyrosine kinases comprising four known family members: JAK1, JAK2, JAK3, and tyrosine kinase 2 (TYK2). JAK1, JAK2, and TYK2 are widely distributed across various cell types and tissues, while JAK3 is primarily expressed in hematopoietic cells, particularly myeloid cells and lymphoid cells. The JAK family transmits downstream intracellular signals generated by over 50 different immunomodulators (including cytokines, interferons, and hormones) to regulate a variety of cellular functions. When cytokine receptors bind to type I or II cytokines, a conformational change occurs, and JAKs bind to these receptors, undergoing autophosphorylation and transphosphorylation, further activating signal transducer and activator of transcription (STAT). Activated STAT enters the nucleus, regulating the transcription of various genes, thereby modulating multiple cellular functions, such as apoptosis, proliferation, migration, maturation, and differentiation of T cells, B cells, natural killer cells, macrophages, and epidermal cells. It also regulates the increased production and release of pro-inflammatory cytokines.

[0005] Different molecules in 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 (γc), activating downstream STAT5 or STAT6. Blocking JAK1 or JAK3 can block the effects of these cytokines. IL-6 depends on JAK1-mediated signal transduction. IL-3, IL-5, erythropoietin, and granulocyte-macrophage colony-stimulating factor (GM-CSF) are mediated by JAK2. TYK2 mediates the signal transduction of IL-12 and IL-23, activating STAT3 or STAT4.

[0006] Multiple studies have shown that the JAK family is involved in the development of various diseases. Loss-of-function mutations in human JAK1 lead to immunodeficiency and pathogen infection, while highly active mutations can cause immune dysfunction or eosinophilic syndrome. Highly active mutants of human JAK2 are associated with myelodysplastic syndrome, leukemia, and lymphoma; loss-of-function mutations in humans have not yet been reported. JAK2 deficiency (JAK2- / -) in mice results in death due to defective erythropoiesis between day 12 and day 13 post-conception. JAK3 deficiency in humans has been described, manifesting as severe comprehensive immunodeficiency in the first few months of life, lacking circulating T cells and NK cells, and exhibiting B cell dysfunction, leading to developmental arrest, severe and recurrent infections, thrush, and diarrhea. Highly active mutations in human JAK3 can cause leukemia. Low TYK2 activity in humans reduces susceptibility to autoimmune diseases.

[0007] JAK inhibitors, such as upadacitinib and abrocitinib, are already approved for the treatment of autoimmune diseases, including rheumatoid arthritis, psoriatic arthritis, ulcerative colitis, and Crohn's disease. The TYK2 selective inhibitor deucravacitinib has also received FDA approval. However, no JAK inhibitors have yet been approved for neurodegenerative diseases, such as Alzheimer's disease and Parkinson's syndrome. Studies show that inflammation plays a crucial role in neurodegenerative diseases, including Alzheimer's, Parkinson's, multiple sclerosis, and amyotrophic lateral sclerosis. Therefore, developing novel JAK1 / TYK2 inhibitors has significant clinical value. Summary of the Invention

[0008] This invention provides compounds of formula (II), their stereoisomers, or pharmaceutically acceptable salts thereof.

[0009]

[0010] in,

[0011] It consists of double or single bonds, and the five-membered ring containing T1, T2, and T3 is an aromatic ring;

[0012] Ring A is selected from ring A1 and ring A2;

[0013] Ring A1 is arbitrarily selected by one or more R a1 Substituted monocyclic 4-8 membered oxoheterocyclic alkyl groups;

[0014] Ring A2 is Where 1 represents the linking site with imidazole N, and 2 represents the linking site with R4;

[0015] Ring B is selected from one or more R. b Substituted monocyclic 4-8 membered nitrogen-containing heterocyclic alkyl groups;

[0016] Ring C is joined with ring B, and ring C is selected from one or more R's. c Substituted 5-10 nitrogen-containing heteroaryl groups;

[0017] T is N, wherein N is optionally quaternized or oxidized;

[0018] When ring A is ring A1, T1 is CR3, T2 is CR3, and T3 is O or NH;

[0019] 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 respectively;

[0020] R1 is -L1-R 11 ;

[0021] L1 is selected from key, -CONR 12 -CO-, O, S, NR 12 and can be 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 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;

[0022] R 11 Selected from H, D, F, Cl, Br, I, CN, and can be 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 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;

[0023] 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 3-8Cycloalkyl and 3-8 membered heterocyclic alkyl groups;

[0024] 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;

[0025] 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;

[0026] 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;

[0027] 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 Cycloalkyl, 3-8 membered heterocycloalkyl, phenyl and 5-6 membered heteroaryl, wherein C 1-4 Alkyl, C 2-4 alkenyl, C 2-4 alkynyl group, C 1-4 Alkoxy, C 1-4 Alkylthio, C 3-8 Cycloalkyl, 3-8 membered heterocycloalkyl, phenyl, and 5-6 membered heteroaryl groups are substituted with CN, and further optionally with one or more R groups. 4a replace;

[0028] Each R a1 Each R b Each R c Each R1a R 11a R 11b R 11c R 11d Each R 2a Each R 3a Each R 3b Each R 4a The following groups are independently selected from H, D, F, Cl, Br, I, OH, NH2, CN, =O, and optionally substituted with one or more R groups: C 1-4 Alkyl, C 2-4 alkenyl, C 2-4 alkynyl group, C 1-4 Alkoxy, C 1-4 Alkylthio, C 3-8 Cycloalkyl, 3-8 membered heterocycloalkyl, phenyl and 5-6 membered heteroaryl;

[0029] Or 2 Rs a1 Connected together, or two Rs b Connected together, or two Rs c Connected together, or R b and R c Linked together, they independently form the following groups optionally substituted with one or more R: C 3-8 Cycloalkyl, 4-8 membered heterocycloalkyl, phenyl or 5-6 membered heteroaryl;

[0030] Each R is independently selected from H, D, F, Cl, Br, I, OH, NH2, CN, and the following groups optionally substituted by one or more F: 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.

[0031] The present invention also provides the compound of formula (I), its stereoisomers or pharmaceutically acceptable salts thereof,

[0032]

[0033] in,

[0034] It consists of double or single bonds, and the five-membered ring containing T1, T2, and T3 is an aromatic ring;

[0035] Ring A is selected from ring A1 and ring A2;

[0036] Ring A1 is arbitrarily selected by one or more Ra1 Substituted monocyclic 4-8 membered oxoheterocyclic alkyl groups;

[0037] Ring A2 is Where 1 represents the linking site with imidazole N, and 2 represents the linking site with R4;

[0038] Ring B is selected from one or more R. b Substituted monocyclic 4-8 membered nitrogen-containing heterocyclic alkyl groups;

[0039] Ring C is joined with ring B, and ring C is selected from one or more R's. c Substituted 5-10 nitrogen-containing heteroaryl groups;

[0040] When ring A is ring A1, T1 is CR3, T2 is CR3, and T3 is O or NH;

[0041] 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 respectively;

[0042] R1 is -L1-R 11 ;

[0043] L1 is selected from key, -CONR 12 -CO-, O, S, NR 12 and can be 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 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;

[0044] R 11 Selected from H, F, Cl, Br, I, CN and optionally one or more R 1a 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;

[0045] 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, C2-4 alkynyl group, C 3-8 Cycloalkyl and 3-8 membered heterocyclic alkyl groups;

[0046] R2 is selected from H, 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;

[0047] R3 is selected from H, 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;

[0048] 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;

[0049] 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 Cycloalkyl, 3-8 membered heterocycloalkyl, phenyl and 5-6 membered heteroaryl, wherein C 1-4 Alkyl, C 2-4 alkenyl, C 2-4 alkynyl group, C 1-4 Alkoxy, C 1-4 Alkylthio, C 3-8 Cycloalkyl, 3-8 membered heterocycloalkyl, phenyl, and 5-6 membered heteroaryl groups are substituted with CN, and further optionally with one or more R groups. 4a replace;

[0050] Each R a1 Each Rb Each R c Each R 1a Each R 2a Each R 3a Each R 3b Each R 4a The following groups are independently selected from H, F, Cl, Br, I, OH, NH2, CN, =O, and optionally substituted with one or more R groups: C 1-4 Alkyl, C 2-4 alkenyl, C 2-4 alkynyl group, C 1-4 Alkoxy, C 1-4 Alkylthio, C 3-8 Cycloalkyl, 3-8 membered heterocycloalkyl, phenyl and 5-6 membered heteroaryl;

[0051] Or 2 Rs a1 Connected together, or two Rs b Connected together, or two Rs c Connected together, or R b and R c Linked together, they independently form the following groups optionally substituted with one or more R: C 3-8 Cycloalkyl, 4-8 membered heterocycloalkyl, phenyl or 5-6 membered heteroaryl;

[0052] Each R is independently selected from H, F, Cl, Br, I, OH, NH2, CN, and the following groups optionally substituted by one or more F: 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.

[0053] In some technical solutions of the present invention, each of the above R is independently selected from H, D, F, Cl, OH, NH2, CN, CH3 and CF3, and other variables are as defined in the present invention.

[0054] In some technical solutions of the present invention, each of the above R is independently selected from H, F, Cl, OH, NH2, CN, CH3 and CF3, and other variables are as defined in the present invention.

[0055] In some technical solutions of the present invention, each of the above R is independently selected from F, and other variables are as defined in the present invention.

[0056] In some technical solutions of the present invention, the above-mentioned R a1The components are independently selected from H, D, F, Cl, Br, I, OH, NH2, CN, and CH3, CH2CH3, vinyl, ethynyl, OCH3, OCH2CH3, cyclopropyl, and cyclobutyl, which are optionally substituted with one or more Rs. Other variables are as defined in this invention.

[0057] In some technical solutions of the present invention, the above-mentioned R a1 The components are independently selected from H, F, Cl, Br, I, OH, NH2, CN, and CH3, CH2CH3, vinyl, ethynyl, OCH3, OCH2CH3, cyclopropyl, and cyclobutyl, which are optionally substituted with one or more Rs, and other variables are as defined in this invention.

[0058] In some technical solutions of the present invention, the above-mentioned R a1 The variables are independently selected from H, D, F, Cl, CH3 and CF3, respectively, and other variables are as defined in this invention.

[0059] In some technical solutions of the present invention, the above-mentioned R a1 The variables are independently selected from H, F, Cl, CH3 and CF3, respectively, and other variables are as defined in this invention.

[0060] In some technical solutions of the present invention, the above two Rs a1 Linked together to form the following groups optionally substituted with one or more R: cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, oxacyclobutyl or oxacyclopentyl, other variables as defined in this invention.

[0061] In some technical solutions of the present invention, the above two Rs a1 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.

[0062] In some technical solutions of the present invention, the above-mentioned R b 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.

[0063] 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.

[0064] In some technical solutions of the present invention, the above-mentioned R bThe 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.

[0065] 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.

[0066] 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.

[0067] 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.

[0068] 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.

[0069] In some technical solutions of the present invention, the above-mentioned R c 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.

[0070] In some technical solutions of the present invention, the above-mentioned R c The variables are independently selected from H, F, Cl, CH3 and CF3, respectively, and other variables are as defined in this invention.

[0071] In some technical solutions of the present invention, the above-mentioned R 1a R 11a R11b R 11c R 11d Each R 2a Each R 3a Each R 3b Each R 4a 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.

[0072] In some technical solutions of the present invention, the above-mentioned R 1a R 11a R 11b R 11c R 11d Each R 2a Each R 3a Each R 3b Each R 4a The components are independently selected from H, D, F, Cl, CN, =O, CH3, CFH2, CF2H, CF3, OCH3, OCF3, and cyclopropyl, respectively, and other variables are as defined in this invention.

[0073] In some technical solutions of the present invention, the above-mentioned R 1a Each R 2a Each R 3a Each R 3b Each R 4a 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.

[0074] In some technical solutions of the present invention, the above-mentioned R 1a Each R 2a Each R 3a Each R 3b Each R 4a The components are independently selected from H, F, Cl, CN, =O, CH3, CFH2, CF2H, CF3, OCH3, OCF3, and cyclopropyl, respectively, and other variables are as defined in this invention.

[0075] In some technical solutions of the present invention, L1 is selected from bonds, C(O)NH, C(O), O, S, NH and optionally bonded by one or more R 1a The following groups may be substituted: CH2, CH2CH2, vinyl, ethynyl, propynyl, OCH2, OCH2CH2 and cyclopropyl, and other variables as defined in this invention.

[0076] In some technical solutions of the present invention, L1 is selected from bonds, C(O), O, S, and optionally R. 1a The following groups may be substituted: C(O)NH, NH, CH2, CH2CH2, vinyl, ethynyl, propynyl, OCH2, OCH2CH2 and cyclopropyl, and other variables as defined in this invention.

[0077] In some technical solutions of the present invention, the L1 mentioned above is selected from the following groups: bond, O, S, and optionally substituted by one or more F or D: CH2, CH2CH2, OCH2, OCH2CH2 and cyclopropyl.

[0078] In some technical solutions of the present invention, the above-mentioned R 11 Selected from H, D, F, Cl, Br, I, CN and optionally one or more R 1a The following groups are 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, oxecyclopentyl, Other variables are as defined in this invention.

[0079] 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.

[0080] In some technical solutions of the present invention, the above-mentioned R 11The 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.

[0081] 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.

[0082] 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.

[0083] In some technical solutions of the present invention, 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, and other variables are as defined in the present invention.

[0084] In some technical solutions of the present invention, L1 is selected from bonds, O and S, R 11 Selected from F, Cl, Br, I, CN, and optionally by one or more R 1a The following groups are substituted: C 1-4 Alkyl, C3-6 Cycloalkyl and 3-6 membered heterocyclic alkyl groups, other variables as defined in this invention.

[0085] In some technical solutions of this invention, R1 is selected from CN, CH3, CFH2, CF2H, CF3, CD3, OCH3, OCF3, OCD3, etc.

[0086] Other variables are as defined in this invention.

[0087] In some technical solutions of the present invention, R1 is selected from... Other variables are as defined in this invention.

[0088] In some technical solutions of the present invention, the R2 is selected from H, D, F, Cl, Br, I, OH, NH2, CN, and optionally one or more R 2a The following groups may be substituted: CH3, CH2CH3, CH2CH2CH3, CH(CH3)2, vinyl, ethynyl, propynyl, OCH3, OCH2CH3, OCH(CH3)2, SCH3, SCH2CH3 and cyclopropyl, and other variables as defined in this invention.

[0089] In some technical solutions of the present invention, R2 is selected from H, D, F, Cl, Br, CN, CH3, CF3, ethynyl and cyclopropyl, and other variables are as defined in the present invention.

[0090] In some technical solutions of the present invention, the R2 is selected from H, F, Cl, Br, I, OH, NH2, CN, and optionally one or more R 2a The following groups may be substituted: CH3, CH2CH3, CH2CH2CH3, CH(CH3)2, vinyl, ethynyl, propynyl, OCH3, OCH2CH3, OCH(CH3)2, SCH3, SCH2CH3 and cyclopropyl, and other variables as defined in this invention.

[0091] In some technical solutions of the present invention, R2 is selected from H, F, Cl, Br, CN, CH3, CF3, ethynyl and cyclopropyl, and other variables are as defined in the present invention.

[0092] In some technical solutions of the present invention, R2 is selected from H, F, Cl, CH3 and CF3, and other variables are as defined in the present invention.

[0093] In some technical solutions of the present invention, R2 is selected from H, and other variables are as defined in the present invention.

[0094] In some technical solutions of the present invention, the R3 is selected from H, D, F, Cl, Br, I, OH, NH2, CN, and optionally one or more R 3a 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, cyclopentyl, cyclohexyl, oxacyclobutyl, aziroxybutyl, phenyl, pyridinyl, pyrazinyl, pyridazinyl, pyrroleyl, pyrazolyl, imidazolyl, furanyl, thiopheneyl, and thiazolyl, with other variables as defined in this invention.

[0095] In some technical solutions of the present invention, R3 is selected from H, D, F, Cl, Br, CN, CH3, CF3, ethynyl and cyclopropyl, and other variables are as defined in the present invention.

[0096] In some technical solutions of the present invention, the R3 is selected from H, F, Cl, Br, I, OH, NH2, CN, and optionally one or more R 3a 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, cyclopentyl, cyclohexyl, oxacyclobutyl, aziroxybutyl, phenyl, pyridinyl, pyrazinyl, pyridazinyl, pyrroleyl, pyrazolyl, imidazolyl, furanyl, thiopheneyl, and thiazolyl, with other variables as defined in this invention.

[0097] In some technical solutions of the present invention, R3 is selected from H, F, Cl, Br, CN, CH3, CF3, ethynyl and cyclopropyl, and other variables are as defined in the present invention.

[0098] In some technical solutions of the present invention, R3 is selected from H, and other variables are as defined in the present invention.

[0099] In some technical solutions of this invention, R4 is selected from CN, CH3, CH2CH3, CH2CH2CH3, CH(CH3)2, vinyl, propynyl, ethynyl, propynyl, OCH3, OCH2CH3, OCH(CH3)2, SCH3, SCH2CH3, cyclopropyl, and cyclobutyl, wherein the CH3, CH2CH3, CH2CH2CH3, CH(CH3)2, vinyl, propynyl, ethynyl, propynyl, OCH3, OCH2CH3, OCH(CH3)2, SCH3, SCH2CH3, cyclopropyl, and cyclobutyl are substituted with CN, and are further optionally substituted with one or more R. 4a Replacement, other variables as defined in this invention.

[0100] In some technical solutions of this invention, R4 is selected from CN, -CH2CN, -C(CH3)2CN, Other variables are as defined in this invention.

[0101] In some technical solutions of the present invention, the ring A1 is selected from one or more R. a1 The substituted oxecyclobutyl, oxecyclopentyl, oxecyclohexyl, oxecycloheptyl, and oxecyclohexyl, and other variables as defined in this invention.

[0102] In some technical solutions of the present invention, the aforementioned ring A1 is selected from... Other variables are as defined in this invention.

[0103] In some technical solutions of the present invention, the ring B is selected from one or more R. b The substituted pyrrolidinyl, piperidinyl, or homopiperidinyl groups, with the ring C selected optionally by one or more R groups, are also present. c The substituted imidazole, pyrazol, pyrrole, pyridazinyl, and pyrimidinyl groups, and other variables as defined in this invention.

[0104] In some technical solutions of the present invention, the aforementioned ring A2 is selected from... Other variables are as defined in this invention.

[0105] In some technical solutions of the present invention, the aforementioned ring A2 is selected from... Other variables are as defined in this invention.

[0106] In some technical solutions of the present invention, the aforementioned ring A2 is selected from... Other variables are as defined in this invention.

[0107] In some technical solutions of the present invention, the above-mentioned structural unit Selected from Other variables are as defined in this invention.

[0108] In some technical solutions of the present invention, the above-mentioned structural unit Selected from Other variables are as defined in this invention.

[0109] In some technical solutions of the present invention, the above-mentioned structural unit Selected from Other variables are as defined in this invention.

[0110] In some technical solutions of the present invention, the above-mentioned structural unit for Other variables are as defined in this invention.

[0111] 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:

[0112]

[0113] in,

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

[0115] R1, R2, R3, R4, R a1 Rings B and C are as defined in this invention.

[0116] 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:

[0117]

[0118] in,

[0119] m and n are independently selected from 0, 1, 2 and 3 respectively;

[0120] R1 is -L1-R 11 ;

[0121] L1 is selected from the bond, -CONH, -CO-, O, S, NH, and optionally 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 1-4 Alkoxy, C 1-4 Alkylthio, C 1-4 Alkylamino, C 3-8 Cycloalkyl and 3-8 membered heterocyclic alkyl groups;

[0122] R 11Selected from H, F, Cl, Br, I, CN and optionally one or more R 1a The following groups are substituted: C 1-4 Alkyl, C 2-4 alkenyl, C 2-4 alkynyl group, C 1-4 Alkoxy, -C 3-8 Cycloalkyl and 3-8 membered heterocyclic alkyl groups;

[0123] R2 is selected from H, 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 3-8 Cycloalkyl and 3-8 membered heterocyclic alkyl groups;

[0124] R3 is selected from H, 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 3-8 Cycloalkyl and 3-8 membered heterocyclic alkyl groups;

[0125] R4 is selected from CN, CH3, CH2CH3, CH2CH2CH3, CH(CH3)2, vinyl, propynyl, ethynyl, propynyl, OCH3, OCH2CH3, OCH(CH3)2, SCH3, SCH2CH3, cyclopropyl, and cyclobutyl, wherein the CH3, CH2CH3, CH2CH2CH3, CH(CH3)2, vinyl, propynyl, ethynyl, propynyl, OCH3, OCH2CH3, OCH(CH3)2, SCH3, SCH2CH3, cyclopropyl, and cyclobutyl are substituted with CN, and are further optionally substituted with one or more R. 4a replace;

[0126] Ring B is selected from one or more R. b Substituted monocyclic 5-7 member nitrogen-containing heterocyclic alkyl groups;

[0127] Ring C is joined with ring B, and ring C is selected from one or more R's. c Substituted 5-6 nitrogen-containing heteroaryl groups;

[0128] R a1 R 2a R 3a R 4a R bR 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.

[0129] 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:

[0130]

[0131] in,

[0132] m is selected from 0, 1, 2, and 3;

[0133] n is selected from 1, 2, and 3;

[0134] 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;

[0135] R2 is selected from H, F, Cl, Br, I, and CH3;

[0136] Each R3 is independently selected from H, F, Cl, Br, I, and CH3;

[0137] R4 is selected from CN, CH2CN, CH2CH2CN, CH(CH3)CN, C(CH3)2CN,

[0138] Each R 1a They were each independently selected from H, D, F, Cl, and CN;

[0139] 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.

[0140] In some technical solutions of the present invention, the compounds of formula (I-1), their stereoisomers, or pharmaceutically acceptable salts thereof are selected from compounds of formula (I-1-1), (I-1-1a), or (I-1-1b); the compounds of formula (I-2), their stereoisomers, or pharmaceutically acceptable salts thereof are selected from compounds of formula (I-2-1), (I-2-1a), or (I-2-1b); and the compounds of formula (I-3), their stereoisomers, or pharmaceutically acceptable salts thereof are selected from compounds of formula (I-3-1), (I-3-1a), or (I-3-1b).

[0141]

[0142] in,

[0143] m and s are independently selected from 0, 1, 2 and 3 respectively;

[0144] R1, R2, R3, R a1 R b R c As defined in this invention.

[0145] 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:

[0146]

[0147] Wherein, R1 is as defined in this invention.

[0148] 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:

[0149]

[0150] in,

[0151] R1 is -L1-R 11 ;

[0152] L1 is selected from bonds, O, and S;

[0153] R 11 Selected from F, Cl, Br, I, CN, and optionally by one or more R 1a The following groups are substituted: C 1-4 Alkyl, C 3-6 Cycloalkyl and 3-6 membered heterocyclic alkyl groups;

[0154] Each R 1a The 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.

[0155] 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:

[0156]

[0157] in,

[0158] 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;

[0159] Each R 1a They were each independently selected from H, D, F, Cl, and CN;

[0160] R4 is selected from CN, CH2CN, CH2CH2CN, CH(CH3)CN, C(CH3)2CN,

[0161] 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.

[0162] 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:

[0163]

[0164] in,

[0165] 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;

[0166] Each R 1a They were each independently selected from H, D, F, Cl, and CN;

[0167] R4 is selected from CN, CH2CN, CH2CH2CN, CH(CH3)CN, C(CH3)2CN,

[0168] In some technical solutions of the present invention, the compounds shown in formulas (I), (I-1), (II-1), (II-1a), (II-1b), (II-1c), (II-1d), (III-1), (III-1a), (III-1b), (III-1c), and (III-1d), their stereoisomers, or pharmaceutically acceptable salts thereof, wherein R1 is selected from F, Cl, Br, I, CN, and CH3 or OCH3 optionally substituted with 1, 2, or 3 F or D atoms;

[0169] In some technical solutions of the present invention, the compounds shown in formulas (I), (I-1), (II-1), (II-1a), (II-1b), (II-1c), (II-1d), (III-1), (III-1a), (III-1b), (III-1c), and (III-1d), their stereoisomers, or pharmaceutically acceptable salts thereof, wherein R4 is selected from CH2CN, and other variables are as defined in the present invention.

[0170] In some technical solutions of the present invention, the compound shown in formula (II), its stereoisomer, or a pharmaceutically acceptable salt thereof is selected from:

[0171]

[0172] Among them, rings A, L1, T, T1, T2, T3, and R 11a R 11b R 11c R4 and As defined in this invention.

[0173] In some technical solutions of the present invention, the compound shown in formula (II), its stereoisomer, or a pharmaceutically acceptable salt thereof is selected from:

[0174]

[0175] in,

[0176] L1 is selected from the following groups: bond, O, S, and optionally substituted by one or more F or D: CH2, CH2CH2, OCH2, OCH2CH2, and cyclopropyl;

[0177] R 11a R 11b R 11c R 11d The following groups are selected independently from H, D, F, Cl, Br, I, OH, NH2, CN, and optionally substituted by one or more F or D groups: C 1-3 Alkyl, C 1-3 Alkoxy, C 1-3 Alkylthio, C 3-6 Cycloalkyl and 3-6 membered heterocyclic alkyl groups;

[0178] Each R a1 They are independently selected from H, D, F, Cl, CH3 and CF3, respectively;

[0179] R2 is selected from H, D, F, Cl, Br, CN, CH3, CF3, ethynyl, and cyclopropyl;

[0180] R3 is selected from H, D, F, Cl, Br, CN, CH3, CF3, ethynyl, and cyclopropyl;

[0181] R4 is selected from CN, C 1-3 Alkyl and C 3-6 cycloalkyl, the C 1-3 Alkyl and C 3-6 Each cycloalkyl group is independently and optionally substituted with CN, and further optionally substituted with one or more F or D;

[0182] Rings B and C are as defined in this invention.

[0183] Some technical solutions of this invention are derived from arbitrary combinations of the above-mentioned variables.

[0184] The present invention also provides the compounds shown in Table A, their stereoisomers, or pharmaceutically acceptable salts thereof.

[0185] In some technical solutions of the present invention, the compounds in Table A, their stereoisomers, or pharmaceutically acceptable salts thereof are selected from the compounds in Table A1.

[0186] Table A Compounds

[0187]

[0188]

[0189] Table A1 Compounds

[0190]

[0191]

[0192]

[0193]

[0194]

[0195]

[0196] The present invention also provides crystal form A of compound 3.

[0197]

[0198] The X-ray powder diffraction pattern of Cu Kα radiation of the A crystal type has characteristic diffraction peaks at the following 2θ angles: 9.01±0.20°, 13.27±0.20° and 22.37±0.20°.

[0199] In some embodiments of the present invention, the X-ray powder diffraction pattern of Cu Kα radiation of the above-mentioned A-type crystal, represented by the 2θ angle, contains at least 5, 6, 7 or 8 diffraction peaks selected from the following: 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°.

[0200] In some technical solutions of the present invention, the X-ray powder diffraction pattern of Cu Kα radiation of the above-mentioned A crystal form 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°.

[0201] In some embodiments of the present invention, the X-ray powder diffraction pattern of Cu Kα radiation of the above-mentioned A-type crystal 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°.

[0202] In some embodiments of the present invention, the X-ray powder diffraction pattern of Cu Kα radiation of the above-mentioned A-type crystal, represented by the 2θ angle, contains at least 10, 11, 12, or 13 diffraction peaks selected from the following: 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°.

[0203] In some embodiments of the present invention, the X-ray powder diffraction pattern of Cu Kα radiation of the above-mentioned A-type crystal has characteristic diffraction peaks at the following 2θ angles: 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°.

[0204] In some embodiments of the present invention, the X-ray powder diffraction pattern of Cu Kα radiation of the above-mentioned A-type crystal has characteristic diffraction peaks at the following 2θ angles: 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 0.81±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°.

[0205] In some embodiments of the present invention, the X-ray powder diffraction pattern of Cu Kα radiation of the above-mentioned A-type crystal has characteristic diffraction peaks at the following 2θ angles: 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°.

[0206] In some embodiments of the present invention, the X-ray powder diffraction pattern of Cu Kα radiation of the above-mentioned A-type crystal has characteristic diffraction peaks at the following 2θ angles: 9.01°, 10.00°, 10.62°, 11.65°, 12.28°, 12.82°, 13.27°, 14.44°, 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°, 26.70°, 27.19°, 29.88°, 30.81°, and 41.16°.

[0207] In some embodiments of the present invention, the X-ray powder diffraction pattern of Cu Kα radiation of the above-mentioned A-type crystal is essentially as follows: Figure 3 As shown.

[0208] 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 A-type crystal are shown in Table 1.

[0209] Table 1: XRPD spectrum analysis data of crystal form A of compound 3

[0210]

[0211] In some technical solutions of the present invention, the differential scanning calorimetry (DSC) curve of the above-mentioned A crystal form has an initial value of endothermic peak at 248.2±3℃.

[0212] In some technical solutions of the present invention, the differential scanning calorimetry (DSC) curve of the above-mentioned A crystal form has an initial value of exothermic peak at 181.3±3℃ and an initial value of endothermic peak at 248.2℃±3℃.

[0213] In some technical solutions of the present invention, the DSC spectrum of the above-mentioned A crystal form is basically as follows: Figure 4 As shown.

[0214] In some technical solutions of the present invention, the thermogravimetric analysis (TGA) curve of the above-mentioned A crystal form shows a weight loss of 0.75% in the range of 30.0±3℃ to 220.0±3℃.

[0215] In some technical solutions of the present invention, the thermogravimetric analysis (TGA) curve of the above-mentioned A crystal form shows a weight loss of 0.33% in the range of 30.0±3℃ to 165.0±3℃, and a further weight loss of 0.42% in the range of 165.0℃±3℃ to 220.0℃±3℃.

[0216] In some technical solutions of the present invention, the TGA spectrum of the above-mentioned A crystal form is basically as follows: Figure 5 As shown.

[0217] The present invention also provides a B-type of compound 3, wherein the Cu Kα radiation X-ray powder diffraction pattern of the B-type has characteristic diffraction peaks at the following 2θ angles: 8.45±0.20°, 9.98±0.20° and 13.60±0.20°.

[0218] In some technical solutions of the present invention, the X-ray powder diffraction pattern of Cu Kα radiation of the above-mentioned B crystal form has 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°.

[0219] In some embodiments of the present invention, the X-ray powder diffraction pattern of Cu Kα radiation of the above-mentioned B crystal form, represented by the 2θ angle, contains at least 6, 7, or 8 diffraction peaks selected from the following: 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°.

[0220] In some embodiments of the present invention, the X-ray powder diffraction pattern of Cu Kα radiation of the above-mentioned B crystal form has 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°.

[0221] In some embodiments of the present invention, the X-ray powder diffraction pattern of Cu Kα radiation of the above-mentioned B crystal form, represented by the 2θ angle, contains at least 10, 11, 12, or 13 diffraction peaks selected from the following: 8.45±0.20°, 9.98±0.20°, 11.88±0.20°, 13.60±0.20°, 16.91±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°, 25.43±0.20°, and 28.89±0.20°.

[0222] In some embodiments of the present invention, the X-ray powder diffraction pattern of Cu Kα radiation of the above-mentioned B crystal form 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°.

[0223] In some embodiments of the present invention, the X-ray powder diffraction pattern of Cu Kα radiation of the above-mentioned B crystal form 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°.

[0224] In some embodiments of the present invention, the X-ray powder diffraction pattern of Cu Kα radiation of the above-mentioned B crystal form has characteristic 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°, 30.50°, and 30.68°.

[0225] In some embodiments of the present invention, the X-ray powder diffraction pattern of Cu Kα radiation of the above-mentioned B-type crystal has characteristic diffraction 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°, 22.1°. 5°, 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°.

[0226] In some embodiments of the present invention, the X-ray powder diffraction pattern of Cu Kα radiation of the above-mentioned B-type crystal is essentially as follows: Figure 6 As shown.

[0227] 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 B crystal form are shown in Table 2.

[0228] Table 2: XRPD spectrum analysis data of crystal form B of compound 3

[0229]

[0230] In some technical solutions of the present invention, the differential scanning calorimetry (DSC) curve of the B crystal form has an initial value of endothermic peak at 249.5±3℃.

[0231] In some technical solutions of the present invention, the DSC spectrum of the above-mentioned B crystal form is basically as follows: Figure 7 As shown.

[0232] In some technical solutions of the present invention, the thermogravimetric analysis (TGA) curve of the above-mentioned B crystal form shows a weight loss of 0.25% in the range of 30.0±3℃ to 150.0±3℃.

[0233] In some technical solutions of the present invention, the TGA spectrum of the above-mentioned B crystal form is basically as follows: Figure 8 As shown.

[0234] The present invention also provides the C crystal form of compound 3, wherein the X-ray powder diffraction pattern of Cu Kα radiation of the C crystal form has characteristic diffraction 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°.

[0235] In some embodiments of the present invention, the X-ray powder diffraction pattern of Cu Kα radiation of the above-mentioned C crystal form, represented by the 2θ angle, contains at least 6, 7, or 8 diffraction peaks selected from the following: 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°.

[0236] In some embodiments of the present invention, the X-ray powder diffraction pattern of Cu Kα radiation of the above-mentioned C crystal form has characteristic diffraction peaks at the following 2θ 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°.

[0237] In some embodiments of the present invention, the X-ray powder diffraction pattern of Cu Kα radiation of the above-mentioned C-type crystal, represented by the 2θ angle, contains at least 10, 11, 12, or 13 diffraction peaks selected from the following: 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°.

[0238] In some embodiments of the present invention, the X-ray powder diffraction pattern of Cu Kα radiation of the above-mentioned C-type crystal has characteristic diffraction peaks at the following 2θ 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°.

[0239] In some embodiments of the present invention, the X-ray powder diffraction pattern of Cu Kα radiation of the above-mentioned C-type crystal is essentially as follows: Figure 9 As shown.

[0240] 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 crystal form are shown in Table 3.

[0241] Table 3: XRPD spectrum analysis data of the C-crystal form of compound 3

[0242]

[0243]

[0244] 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℃.

[0245] In some technical solutions of the present invention, the DSC spectrum of the above-mentioned C crystal form is basically as follows: Figure 10 As shown.

[0246] 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℃.

[0247] In some technical solutions of the present invention, the TGA spectrum of the above-mentioned C-type crystal is basically as follows: Figure 11 As shown.

[0248] The present invention also provides a hydrate of compound 3; preferably, the hydrate of compound 3 is shown in formula 3'.

[0249]

[0250] 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°.

[0251] 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°.

[0252] 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°.

[0253] 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 10, 11, 12, or 13 diffraction peaks selected from the following: 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°.

[0254] 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°, 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°.

[0255] 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°, 10.93°, 11.52°, 13.31°, 13.60°, 14.11°, 15.01°, 15.92°, 16.41°, 16.89°, 17.23°, 17.67°, 18.48°, 18.96°, 20.11°, 20.59°, 20.93°, 21.46°, 21. 87°, 22.22°, 23.19°, 23.49°, 23.78°, 23.98°, 24.22°, 24.68°, 24.93°, 25.25°, 26.21°, 26.88°, 27.54°, 27.92°, 28.29°, 29.25°, 30.14°, 30.89°, 31.36°, 32.29°, 32.94°, 34.05° and 37.34°.

[0256] In some embodiments of the present invention, the X-ray powder diffraction pattern of Cu Kα radiation of the above-mentioned D-type crystal is essentially as follows: Figure 12 As shown.

[0257] 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 D crystal form are shown in Table 4.

[0258] Table 4: XRPD spectrum analysis data of the D crystal form of compound 3

[0259]

[0260] In some technical solutions of the present invention, the differential scanning calorimetry (DSC) curve of the above-mentioned D crystal form has an initial value of endothermic peak at 251.1℃±3℃.

[0261] In some technical solutions of the present invention, the DSC spectrum of the above-mentioned D crystal form is basically as follows: Figure 13 As shown.

[0262] In some technical solutions of the present invention, the thermogravimetric analysis (TGA) curve of the above-mentioned D crystal form shows a weight loss of 5.16% in the range of 30.0℃±3℃ to 120.0℃±3℃.

[0263] In some technical solutions of the present invention, the TGA spectrum of the above-mentioned D crystal form is basically as follows: Figure 14 As shown.

[0264] The crystals of compound 3 described in this invention can be in the form of a nonsolvent or a solvate, such as a hydrate.

[0265] The present invention provides a crystalline composition comprising crystals of compound 3, wherein the crystals of compound 3 constitute 50% or more by weight of the crystalline composition, preferably 75% or more, more preferably 90% or more, and most preferably 95% or more. The crystalline compositions may also contain small amounts of other crystalline or amorphous forms of compound 3.

[0266] The present invention provides a crystalline composition comprising compound 3' crystals, wherein the compound 3' crystals constitute 50% or more by weight of the crystalline composition, preferably 75% or more, more preferably 90% or more, and most preferably 95% or more. The crystalline compositions may also contain small amounts of other crystalline or amorphous forms of compound 3'.

[0267] This invention provides a pharmaceutical composition comprising a therapeutically effective amount of the aforementioned compound, or its crystals, or a crystalline composition thereof; the pharmaceutical composition may contain at least one pharmaceutically acceptable carrier or other excipient. Furthermore, the pharmaceutical composition of this application may further comprise one or more other therapeutic agents.

[0268] The present invention also provides the use of the above-mentioned compounds, their stereoisomers or pharmaceutically acceptable salts thereof in the preparation of medicaments for treating diseases related to JAK1 / TYK2 inhibitors.

[0269] The present invention also provides the use of the above-mentioned compounds, their stereoisomers, or pharmaceutically acceptable salts thereof, or crystals thereof, in the preparation of medicaments for treating diseases related to JAK1 / TYK2 inhibitors.

[0270] The present invention also provides the use of the above-mentioned compounds, their stereoisomers, or pharmaceutically acceptable salts or B crystal forms in the preparation of medicaments for treating diseases related to JAK1 / TYK2 inhibitors.

[0271] In some technical solutions of the present invention, the diseases related to the above-mentioned JAK1 / TYK2 inhibitors are autoimmune diseases, neuroinflammatory lesions, and neurodegenerative diseases.

[0272] In some technical solutions of the present invention, the aforementioned autoimmune diseases include, but are not limited to, rheumatoid arthritis, vitiligo, Crohn's disease, ulcerative colitis, etc.

[0273] In some technical solutions of the present invention, the aforementioned neuroinflammatory and neurodegenerative diseases include, but are not limited to, Alzheimer's disease, Parkinson's syndrome, multiple sclerosis, and amyotrophic lateral sclerosis.

[0274] In some technical solutions of the present invention, the aforementioned neuroinflammatory and neurodegenerative diseases include, but are not limited to, Alzheimer's disease (and amyloid-related imaging abnormalities after antibody treatment), Parkinson's syndrome, and multiple sclerosis.

[0275] Technical effect

[0276] The compounds of this invention exhibit strong inhibitory activity against both JAK1 and TYK2 kinases, with significantly weaker inhibitory activity against JAK2 and JAK3 than against JAK1, demonstrating high selectivity. They also show strong inhibitory activity against both JAK1 and TYK2 in human peripheral blood mononuclear cells, with weaker inhibition against JAK2, exhibiting high selectivity. Furthermore, they significantly reduce the secretion of inflammatory factors (IL-6, TNF-α, and CCL-2) in a dose-dependent manner in cellular inflammation models. The compounds show high concentrations of free drug in plasmas of different species, indicating good drug-like properties. They exhibit good stability in liver microsomes and hepatocytes of various species. In MDCKII-MDR1 monolayer cell permeability experiments, they demonstrate high permeability and low efflux. They also exhibit good stability in human liver microsomes. It showed no significant inhibitory effects on the five major cytochrome P450 enzymes (CYP1A2, CYP2C9, CYP2C19, CYP2D6, and CYP3A4), indicating a low risk of drug-drug interactions. It also showed no significant induction of CAR, PXR, and AhR, with a very low risk of inducing major cytochrome P450 enzymes. Furthermore, it exhibited no significant inhibition of hERG. In PK experiments across various genera, it demonstrated high oral exposure and high oral bioavailability with minimal species variation, exhibiting excellent pharmacokinetic properties. It also showed good brain tissue distribution and excellent brain penetration. In the EAE pharmacodynamic model, it demonstrated significant therapeutic effects with a low onset dose and a dose-effect positive correlation, showcasing superior in vivo pharmacodynamic properties.

[0277] The crystal form preparation process of the compound of the present invention is simple, its physical and chemical properties are stable, it is slightly hygroscopic, and it has good application prospects.

[0278] Definitions and Explanations

[0279] Unless otherwise stated, the following terms and phrases as used herein are intended to have the following meanings. A particular term or phrase should not be considered uncertain or unclear unless specifically defined, but should be understood in its ordinary sense. When a trade name appears herein, it is intended to refer to the corresponding product or its active ingredient.

[0280] The term “pharmaceutically acceptable” as used herein refers to compounds, materials, compositions, and / or dosage forms that, within the bounds of reliable medical judgment, are suitable for use in contact with human and animal tissues without excessive toxicity, irritation, allergic reactions, or other problems or complications, in proportion to a reasonable benefit / risk ratio.

[0281] The term "pharmaceutically acceptable salt" refers to a salt of the compounds of the present invention, prepared by reacting a compound having specific substituents, as discovered in the present invention, with a relatively non-toxic acid or base. When the compounds of the present invention contain relatively acidic functional groups, base addition salts can be obtained by contacting such compounds with a sufficient amount of base in a pure solution or a suitable inert solvent. When the compounds of the present invention contain relatively basic functional groups, acid addition salts can be obtained by contacting such compounds with a sufficient amount of acid in a pure solution or a suitable inert solvent. The pharmaceutically acceptable salts of the present invention can be synthesized from parent compounds containing acid radicals or bases by conventional chemical methods. Generally, such salts are prepared by reacting these compounds in their free acid or base form with a stoichiometric amount of a suitable base or acid in water or an organic solvent or a mixture of both.

[0282] The compounds of this invention can exist in specific geometric or stereoisomeric forms. This invention envisions all such compounds, including cis and trans isomers, trans-isomers, (-)- and (+)-enantiomers, (R)- and (S)-enantiomers, diastereomers, (D)-isomers, (L)-isomers, and racemic mixtures thereof, as well as other mixtures, such as mixtures enriched with enantiomers or diastereomers, all of which are within the scope of this invention. Additional asymmetric carbon atoms may be present in substituents such as alkyl groups. All such isomers and mixtures thereof are included within the scope of this invention. The optical purity of a single-configuration compound can be expressed as optical rotation, chiral purity, or ee, etc. Chiral purity refers to the content determined by testing methods (such as GC, HPLC, SFC, NMR, etc.); ee refers to the percentage excess of isomers or enantiomers, which is the difference in the percentage content of two isomers or two enantiomers. For example, if SFC testing shows that the content of one isomer a is 90% and the content of another isomer b is 10%, then the chiral purity of isomer a is 90% and the ee value is 80%.

[0283] The compounds of this invention can exist in specific tautomers. Unless otherwise stated, the terms "tautomer" or "tautomer form" refer to functional group isomers resulting from the rapid movement of one or more atoms in a molecule between two positions. A tautomer is a special type of functional group isomer. Tautomers can interconvert and exist in dynamic equilibrium, but usually exist primarily in the more stable isomer form. For example, chemical equilibrium of tautomers can be achieved in solution. For example, proton tautomers (also called prototropic tautomers) include interconversions via proton migration, such as keto-enol isomerization and imine-enamine isomerization. Valence tautomers include interconversions involving the rearrangement of some bonding electrons. For example, a specific example of keto-enol tautomerization is the interconversion between the two tautomers, pentane-2,4-dione and 4-hydroxypent-3-en-2-one.

[0284] Unless otherwise stated, the terms "enantiomer" or "optical isomer" refer to stereoisomers that are mirror images of each other.

[0285] Unless otherwise stated, the terms "cis-trans isomers" or "geometric isomers" arise because the single bonds of double bonds or cyclic carbon atoms cannot rotate freely.

[0286] Unless otherwise stated, the term "diastereomer" refers to a stereoisomer of a molecule having two or more chiral centers and being a non-mirror image of each other. Unless otherwise stated, "(+)" indicates dextrorotatory, "(-)" indicates levorotatory, and "(±)" indicates racemic.

[0287] Unless otherwise specified, use wedge-shaped solid line keys. and wedge-shaped dashed key The absolute configuration of the center of a solid is represented by a straight solid line key. and straight dashed key The relative configuration of the center of a solid is indicated by a wavy line. Indicates wedge-shaped solid line key and / or wedge-shaped dashed key Or use wavy lines Indicates a straight solid line key and / or straight dashed key

[0288] Unless otherwise stated, carbon atoms marked with an asterisk (*) are chiral carbon atoms, existing in a single enantiomer (R) or (S) form or enriched with one enantiomer. For example, express or Or it may contain an enantiomer.

[0289] Unless otherwise stated, the terms "rich in one isomer," "isomer enrichment," "rich in one enantiomer," or "enantiomer enrichment" 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%.

[0290] Optically active (R)- and (S)- isomers, as well as D- and L- isomers, can be prepared by chiral synthesis, chiral reagents, or other conventional techniques. To obtain an enantiomer of a compound of the present invention, it can be prepared by asymmetric synthesis or derivatization with a chiral auxiliary, wherein the resulting diastereomeric mixture is separated, and the auxiliary group is cleaved to provide the desired enantiomer in pure form. Alternatively, when the molecule contains a basic functional group (such as an amino group) or an acidic functional group (such as a carboxyl group), a salt of the diastereomeric isomer is formed with a suitable optically active acid or base, followed by diastereomeric resolution using conventional methods known in the art, and then the pure enantiomer is recovered. Furthermore, the separation of enantiomers and diastereomeric isomers is typically accomplished by using chromatography employing a chiral stationary phase, optionally combined with chemical derivatization (e.g., from amines to carbamates).

[0291] The compounds of this invention may contain atomic isotopes in non-natural proportions on one or more atoms constituting the compound. For example, the compounds may be labeled with radioactive isotopes, such as tritium. 3 H), Iodine-125 125 I) or C-14 14 C). For example, deuterium can be used to replace hydrogen to form deuterated drugs. The bond between deuterium and carbon is stronger than that between ordinary hydrogen and carbon. Compared with undeuterated drugs, deuterated drugs have advantages such as reduced toxicity, increased drug stability, enhanced efficacy, and prolonged drug biological half-life. All isotopic variations of the compounds of this invention, regardless of radioactivity, are included within the scope of this invention.

[0292] The term "substituted" refers to the substitution of one or more hydrogen atoms on a particular atom by a substituent. Substituents can include deuterium and hydrogen variants, provided that the valence state of the particular atom is normal and the resulting compound is stable. When the substituent is oxygen (i.e., =O), it means that two hydrogen atoms are substituted.

[0293] The terms “optional” or “optionally” refer to events or conditions that may occur but are not required to occur as described below, and the description includes both cases where said events or conditions occur and cases where said events or conditions do not occur.

[0294] The term "optionally substituted" means that it may or may not be substituted, unless otherwise specified, and the type and number of substituents may be arbitrary on the basis of chemical feasibility.

[0295] In some technical solutions of the present invention, "optionally replaced by one or more R" means that it may not be replaced, or it may be replaced. The number of substituents R is 1 to 10, for example, it may be replaced by 1, 2, 3, 4, 5, 6, 7 or 8 R, or by 1, 2, 3, 4 or 5 R, or by 1, 2 or 3 R. When it is replaced by multiple R, each R may be the same or different.

[0296] When any variable (e.g., R) appears more than once in the composition or structure of a compound, its definition is independent in each case. Thus, for example, if a group is substituted by 0-2 Rs, the group can optionally be substituted by at most two Rs, and the Rs in each case have independent options. Furthermore, combinations of substituents and / or their variants are only permitted if such combinations produce a stable compound.

[0297] When the number of a linking group is 0, such as -(CRR)0-, it indicates that the linking group is a single bond.

[0298] When one of the variables is selected as a single bond, it means that the two groups it connects to are directly connected. For example, when L in ALZ represents a single bond, it means that the structure is actually AZ.

[0299] When a substituent is vacant, it means that the substituent does not exist. For example, if X is vacant in AX, it means that the structure is actually A.

[0300] When the listed substituents do not specify which atom they are attached to the substituted group, they can be bonded to any of their atoms. For example, a pyridinium group, as a substituent, can be attached to the substituted group via any carbon atom on the pyridine ring. When the listed linking groups do not specify their attachment direction, the attachment direction is arbitrary. For example, The linker group L is -MW-. In this case, -MW- can connect ring A and ring B in the same direction as the reading order from left to right to form a ring. Alternatively, rings A and B can be connected in the opposite direction to the right-to-left reading order to form a ring. The combination of linking groups, substituents, and / or their variants is permitted only if such a combination produces a stable compound.

[0301] Unless otherwise specified, when a group has one or more connectable sites, any one or more sites of that group can be connected to other groups by chemical bonds. When the chemical bond connection is non-directional and the connectable site contains H atoms, the number of H atoms at that site will decrease accordingly with the number of chemical bonds connected, resulting in a group with a corresponding valence. The chemical bonds connecting the site to other groups can be straight solid line bonds. Straight dashed key or wavy line Indicated. Wherein, the key is represented by a straight dashed line. or wavy line When indicating a linking site, it can be a single bond, double bond, or triple bond, etc. For example, the straight solid line bond in -OCH3 indicates that the oxygen atom in this group is connected to other groups; The straight dashed bond in the diagram indicates that the group is connected to other groups through both ends of the nitrogen atom in the group; The wavy lines in the text indicate that the phenyl group is connected to other groups through the carbon atoms at positions 1 and 2 of the phenyl group. This indicates that any connectable site on the piperidinyl group can be linked to other groups via a single chemical bond, including at least... Even if H atoms are drawn on -N- in these four connection methods, Still includes In this type of linkage, when a chemical bond is attached, the number of hydrogen atoms at that site is reduced by one, resulting in a monovalent piperidinyl group.

[0302] Unless otherwise specified, C n-n+m Or C n -C n+m This includes any specific case with n to n+m carbons, and also any range from n to n+m. For example, C 1-12 Including C1, C2, C3, C4, C5, C6, C7, C8, C9, C 10 C 11 and C 12 etc., including 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 9-12Similarly, n to n+m membered rings represent rings with n to n+m atoms. For example, 3-12 membered rings include 3-membered, 4-membered, 5-membered, 6-membered, 7-membered, 8-membered, 9-membered, 10-membered, 11-membered, and 12-membered rings, 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.

[0303] Unless otherwise specified, the term “halogen” or “halogen” itself or as part of another substituent means a fluorine, chlorine, bromine or iodine atom.

[0304] Unless otherwise specified, the term "alkyl" on its own or in combination with other terms refers to a straight-chain or branched saturated hydrocarbon group consisting of 1 to 20 carbon atoms. It can be monovalent (e.g., methyl), divalent (e.g., methylene), or polyvalent (e.g., methine). The alkyl group includes C... 1-10 Alkyl, C 1-6 Alkyl, C 1-5 Alkyl, C 1-4 Alkyl, C 1-3 Alkyl groups, examples of which include, but are not limited to, methyl (Me), methylene (CH2), methine (CH), ethyl (Et), propyl (including n-propyl and isopropyl), n-butyl, tert-butyl, n-pentyl, etc. For example, in some technical solutions of the present invention, the alkyl group is C24. 1-6 Alkyl groups, 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 groups, etc.; in other technical solutions of the present invention, the alkyl group is C1, C2, C3, C4, C5, C6 alkyl groups, etc. 1-4 Alkyl groups, including C 1-2 C 1-3 C 2-3 C 2-4 C1, C2, C3, C4 alkyl groups, etc.; in other technical solutions of the present invention, the alkyl group is C1, C2, C3, C4 alkyl group, etc. 1-3 Alkyl groups, including C 1-2 C 2-3 C1, C2, C3 alkyl groups, etc.

[0305] Unless otherwise specified, the term "alkenyl" 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 double bond. It can be monovalent, divalent, or polyvalent. 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.

[0306] 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.

[0307] 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 1-2C 1-3 C 1-4 C 2-3 C 2-4 C 2-5 C1, C2, C3, C4, C5, C6 alkoxy groups, etc.; in some other technical solutions of the present invention, the alkoxy group is C 1-4 Alkoxy groups, which include C 1-2 C 1-3 C 2-3 C 2-4 C1, C2, C3, C4 alkoxy groups, etc.; in some other technical solutions of the present invention, the alkoxy group is C 1-3 Alkoxy groups, which include C 1-2 C 2-3 C1, C2, C3 alkoxy groups, etc.

[0308] Unless otherwise specified, the term "alkathioyl" 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 sulfur atom. It can be monovalent, divalent, or polyvalent. The alkathioyl group includes C... 1-10 Alkylthio, C 1-6 Alkylthio, C 1-5 Alkylthio, C 1-4 Alkylthio, C 1-3 Alkylthio groups, etc., examples of which include, but are not limited to, methylthio, ethylthio, propylthio (including n-propylthio and isopropylthio), etc. For example, in some technical solutions of the present invention, the alkylthio group is C10. 1-6 Alkylthio 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, C6 alkylthio groups, etc.; in some other technical solutions of the present invention, the alkylthio group is C1, C2, C3, C4, C5, C6 alkylthio groups, etc. 1-4 Alkylthio groups, which include C 1-2 C 1-3 C 2-3 C 2-4 C1, C2, C3, C4 alkylthio groups, etc.; in some other technical solutions of the present invention, the alkylthio group is C 1-3 Alkylthio groups, which include C 1-2 C 2-3 C1, C2, C3 alkylthio groups, etc.

[0309] 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.

[0310] 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 groups, etc., examples of which include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cyclohexadienyl, etc. For example, in some technical solutions of the present invention, the cycloalkyl group is C10. 3-6 cycloalkyl groups, including C 3-5 C 4-5 C 4-6 C3, C4, C5, C6 cycloalkyl groups, etc.; in some technical solutions of the present invention, the cycloalkyl group is C 3-8 cycloalkyl groups, including C 3-5 C 4-5 C 4-6 C3, C4, C5, C6, C7, C8 cycloalkyl groups, etc.; in some technical solutions of the present invention, the cycloalkyl group is C 3-10 Cycloalkenyl groups contain at least one carbon-carbon double bond and include 3-5, 4-5, 4-6, 5-6, 5-7, 5-8, 5-9, and 5-10 cycloalkenyl groups.

[0311] Unless otherwise specified, the term "heterocyclic alkyl" on its own or in combination with other terms refers to a saturated or partially unsaturated cyclic group consisting of 3 to 20 ring atoms, wherein 1, 2, 3, 4, 5, 6, 7, or 8 of the ring atoms are heteroatoms independently selected from O, S, and N, and the remainder are carbon atoms, wherein the carbon atoms are optionally oxidized (i.e., C(O)), the nitrogen atom is optionally quaternized, and the nitrogen and sulfur heteroatoms are optionally oxidized (i.e., NO and S(O)2, p is 1 or 2), and the heteroatoms may occupy the connection positions between the heterocyclic alkyl group and the rest of the molecule. It can be monovalent, divalent, or polyvalent. The heterocyclic alkyl group may optionally contain one or more double or triple bonds, but all rings are not aromatic rings. The heterocyclic alkyl group may be a saturated heterocyclic alkyl group (meaning all rings are saturated), or a heterocyclic alkenyl group (meaning a monocyclic or polycyclic system containing at least one carbon-carbon double bond), etc. The heterocyclic alkyl group includes 3-10 membered heterocyclic alkyl groups, 3-8 membered heterocyclic alkyl groups, 3-7 membered heterocyclic alkyl groups, 3-6 membered heterocyclic alkyl groups, 3-5 membered heterocyclic alkyl groups, 4-6 membered heterocyclic alkyl groups, 5-8 membered heterocyclic alkyl groups, 6-8 membered heterocyclic alkyl groups, etc. Examples of heterocyclic alkyl groups include, but are not limited to, nitrogen-containing heterocyclic butyl group, oxo-heterocyclic butyl group, thiocyclic butyl group, pyrrolidinyl group, pyrazolylyl group, imidazoalkyl group, tetrahydrothiophenyl group (including tetrahydrothiophen-2-yl and...). Tetrahydrothiophene-3-yl, etc.), tetrahydrofuranyl (including tetrahydrofuran-2-yl, etc.), tetrahydropyranyl, piperidinyl (including 1-piperidinyl, 2-piperidinyl and 3-piperidinyl, etc.), piperazineyl (including 1-piperidinyl and 2-piperidinyl, etc.), morpholinyl (including 3-morpholinyl and 4-morpholinyl, etc.), dioxane, dithiaalkyl, isoxazolyl, isothiazolyl, 1,2-oxazinyl, 1,2-thiaazinyl, hexahydropyridazinyl For example, in some technical solutions of the present invention, the heterocyclic alkyl group is a 3-6 member heterocyclic alkyl group, including 3-5 member, 4-5 member, 4-6 member, 3 member, 4 member, 5 member, 6 member heterocyclic alkyl groups, etc. The heterocyclic alkyl group is a 3-8 membered heterocyclic alkyl group, including 3-5, 4-5, 4-6, 3, 4, 5, 6, 7, and 8 membered heterocyclic alkyl groups. In some technical solutions of the present invention, the heterocyclic alkyl group is a monocyclic 4-8 membered nitrogen heterocyclic alkyl group, which refers to a saturated or partially unsaturated monocyclic group with 4-8 ring atoms, wherein at least one of the heteroatoms in the ring atoms is an nitrogen atom, including 4-5, 4-6, 5-6, 5-7, and 5-8 membered heterocyclic alkyl groups. In some technical solutions of the present invention, the heterocyclic alkyl group is a monocyclic 4-8 membered oxocyclic alkyl group, which refers to a saturated or partially unsaturated monocyclic group with 4-8 ring atoms, wherein at least one of the heteroatoms in the ring atoms is an oxygen atom, including 4-5, 4-6, 5-6, 5-7, and 5-8 membered heterocyclic alkyl groups.

[0312] Unless otherwise specified, the terms "heteroaryl ring" and "heteroaryl" are used interchangeably. The term "heteroaryl," either alone or in combination with other terms, refers to a monocyclic group or polycyclic ring system consisting of 5 to 20 ring atoms with a conjugated π-electron system, wherein 1, 2, 3, 4, 5, 6, 7, or 8 ring atoms are heteroatoms independently selected from O, S, and N, and the remainder are carbon atoms. The nitrogen atom is optionally quaternized, and the nitrogen and sulfur heteroatoms are optionally oxidized (i.e., NO and S(O)₂, p is 1 or 2). The heteroaryl group can be attached to the rest of the molecule via a heteroatom or a carbon atom, and can be monovalent, divalent, or polyvalent. The heteroaryl groups include 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, and 10-membered heteroaryl groups. Examples of the heteroaryl groups include, but are not limited to, pyrrole (including N-pyrrole, 2-pyrrole, and 3-pyrrole), pyrazolyl (including 2-pyrazolyl and 3-pyrazolyl), imidazole (including N-imidazolyl, 2-imidazolyl, 4-imidazolyl, and 5-imidazolyl), oxazolyl (including 2-oxazolyl, 4-oxazolyl, and 5-oxazolyl), and triazolyl (1H-1,2,3-triazolyl, 2H-1,2,3-triazolyl, 1H-1,2,4-triazolyl, and 4H-1,2,4-triazolyl). (e.g., tetrazolyl, isoxazolyl (3-isooxazolyl, 4-isooxazolyl and 5-isooxazolyl, etc.), thiazolyl (including 2-thiazolyl, 4-thiazolyl and 5-thiazolyl, etc.), furanyl (including 2-furanyl and 3-furanyl, etc.), thienyl (including 2-thienyl and 3-thienyl, etc.), pyridinyl (including 2-pyridinyl, 3-pyridinyl and 4-pyridinyl, etc.), pyrazinyl, pyrazinyl, pyrimidinyl (including 2-pyrimidinyl and 4-pyrimidinyl, etc.), indoleyl, indazoleyl, pyrimidinimidazoleyl, etc. For example, in some technical solutions of the present invention, the heteroaryl group is a 5-10 ternaryl group, which includes 5-6 ternary, 5-8 ternary, 5-9 ternary, 5 ternary, 6 ternary, 7 ternary, 8 ternary, 9 ternary, and 10 ternary heteroaryl groups; in other technical solutions of the present invention, the heteroaryl group is a 5-6 ternary heteroaryl group, which includes 5 ternary and 6 ternary heteroaryl groups.

[0313] Unless otherwise specified, the term "aromatic ring" refers to a cyclic group with a conjugated π-electron system, whose atoms are covered by a delocalized π-electron cloud. In the structural formula, when conforming to the rules of atomic valence and covalent bonding, it can be written in the form of alternating single and double bonds, or it can be written using... This represents the delocalized π-electron cloud. For example, the structural formula... The structures represented are all the same; structural formula The structures represented are all 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.

[0314] 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.

[0315] 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.

[0316] 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.

[0317] Unless otherwise stated, in the differential scanning calorimetry curves of the compounds of the present invention, upward indicates exothermic (Exo Up).

[0318] Unless otherwise stated, room temperature in this invention refers to 15°C to 35°C.

[0319] "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.

[0320] 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.

[0321] The term "therapeutic effective dose" refers to a sufficient amount of a non-toxic drug or agent that achieves the desired effect. The determination of the effective dose varies from person to person, depending on the recipient's age and general condition, as well as the specific active substance. The appropriate effective dose in a case can be determined by a person skilled in the art based on routine testing.

[0322] The term "treatment" means administering the compound or preparation described in this application to improve or eliminate a disease or one or more symptoms associated with the disease, and includes: 1. suppressing the disease or disease state, i.e., curbing its development; 2. alleviating the disease or disease state, even if the disease or disease state subsides.

[0323] In this invention, "pharmaceutically acceptable carriers" refer to carriers that are administered together with the active ingredient, have no significant irritant effect on the organism, and do not impair the biological activity and properties of the active compound.

[0324] The word “comprise” or “include” and its English variants such as comprises or comprising should be understood in an open, non-exclusive sense, meaning “including but not limited to”.

[0325] The structures of the compounds of this invention can be confirmed by conventional methods well known to those skilled in the art. If this invention relates to the absolute configuration of a compound, that absolute configuration can be confirmed by conventional techniques in the art. For example, single-crystal X-ray diffraction (SXRD) is used, where the cultured single crystal is used to collect diffraction intensity data using a Bruker D8 venture diffractometer with CuKα radiation as the light source. The scanning method is as follows: After scanning and collecting relevant data, the crystal structure can be further analyzed using the direct method (Shelxs97) to confirm the absolute configuration.

[0326] Abbreviations of this invention: prep-HPLC represents high performance liquid chromatography for preparation and separation; TLC represents thin-layer chromatography; HPMC represents hydroxypropyl methylcellulose; Saline represents physiological saline; Solutol represents polyethylene glycol-15-hydroxystearate; Tween80 represents Tween 80; PEG400 represents polyethylene glycol 400; HEPES represents 4-hydroxyethylpiperazine ethanesulfonic acid; HBSS represents Hank's balanced salt buffer; NADPH represents reduced coenzyme II, also known as reduced nicotinamide adenine dinucleotide phosphate; PBS represents phosphate buffer.

[0327] The solvents used in this invention are commercially available. Compounds are named according to conventional naming principles in the art or using… Software naming conventions are used; commercially available compounds use supplier catalog names.

[0328] The instrument and analytical method of this invention

[0329] (1) X-ray powder diffractometer (XRPD) method

[0330] Take an appropriate amount of sample powder (approximately 20 mg) into the zero-background sample dish and detect XRPD. Instrument parameters are as follows:

[0331] Instrument Model: BRUKER D8 ADVANCE

[0332] X-ray source: Cu Kα, Kα1: Kα2: The intensity ratio of Kα2 / Kα1 is 0.5.

[0333] Phototube voltage / phototube current: 40kV, 40mA

[0334] Diverging slit: 0.6mm

[0335] Scanning range: 3°~45°

[0336] Scanning mode: Continuous scan

[0337] Step size: 0.02°

[0338] Scan time per step: 0.1s.

[0339] (2) Differential Scanning Calorimeter (DSC) and Thermogravimetric Analysis (TGA)

[0340] The instrument parameters for DSC and TGA tests are shown in Table 5.

[0341] Table 5 DSC and TGA Instrument Parameters

[0342]

[0343] (3) Dynamic Vapor Sorption (DVS) method

[0344] The parameters of the DVS instrument are shown in Table 6.

[0345] Table 6 DVS Instrument Parameters

[0346] Instrument Model SMSDVSintrinsic PLUS temperature 25℃ Sample volume 15-20mg Carrier gas and flow rate <![CDATA[N2,200mL / min]]> dm / dt 0.005% / min Minimum balancing time 10min Maximum balancing time 180min Humidity range 50%RH-0%RH-95%RH-50%RH Humidity gradient 10％(90％-0％-90％)；5％(95％-90％,90％-95％) . Attached Figure Description

[0347] Figure 1.1Clinical scoring in animals in the EAE pharmacodynamic model (Part 1)

[0348] Figure 1.2 Body weight changes in animals in the EAE pharmacodynamic model

[0349] Figure 2 Clinical scoring in animals in the EAE pharmacodynamic model (Part 2)

[0350] Figure 3 XRPD spectrum of crystal form A of compound 3

[0351] Figure 4 DSC spectrum of crystal form A of compound 3

[0352] Figure 5 TGA spectrum of crystal form A of compound 3

[0353] Figure 6 XRPD spectrum of compound 3 in crystal form B

[0354] Figure 7 DSC spectrum of crystal form B of compound 3

[0355] Figure 8 TGA spectrum of compound 3 in crystal form B

[0356] Figure 9 XRPD spectrum of the C-crystal form of compound 3

[0357] Figure 10 DSC spectrum of the C-crystal form of compound 3

[0358] Figure 11 TGA spectrum of the C-crystal form of compound 3

[0359] Figure 12 XRPD spectrum of the D crystal form of compound 3'

[0360] Figure 13 DSC spectrum of the D crystal form of compound 3'

[0361] Figure 14 TGA spectrum of the D crystal form of compound 3'

[0362] Figure 15 DVS spectrum of compound 3 in crystal form B

[0363] Figure 16 DVS spectrum of the D crystal form of compound 3' Detailed Implementation

[0364] The present invention will be described in detail below with reference to embodiments, but this does not imply any adverse limitation on the invention. The present invention has been described in detail, and specific embodiments thereof have been disclosed. It will be apparent to those skilled in the art that various changes and modifications can be made to the specific embodiments of the present invention without departing from the spirit and scope thereof.

[0365] Example 1

[0366]

[0367] Step 1: Under a nitrogen atmosphere, compound 1-1 (1.2 g, 3.41 mmol) and compound 1-2 (574 mg, 4.09 mmol) were added to tert-butanol (12 mL), followed by N,N-diisopropylethylamine (1.54 g, 11.9 mmol). The reaction mixture was heated to 100 °C and reacted for 1 hour. After the reaction was complete, the reaction mixture was cooled to room temperature, diluted with 40 mL of ethyl acetate, and washed successively with water (40 mL × 3) and saturated brine (50 mL). The organic phase was then 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 obtain compounds 1-3A and 1-3B.

[0368] Characterization of compounds 1-3A: TLC developing solvent was petroleum ether:ethyl acetate = 1:1, R f =0.4; LCMS:456.1[M+H] + ; 1 H NMR (400MHz, CDCl3) δ = 9.58 (br d,J=7.6Hz,1H),9.14(s,1H),8.08(d,J=8.4Hz,2H),7.62(d,J=4.0Hz,1H),7.32( d,J=8.0Hz,2H),6.64(d,J=4.0Hz,1H),4.25-4.19(m,1H),4.16-4.12(m,1H),3.83 (dd,J=1.6,12.4Hz,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=4.0,13.2Hz,1H),1.85-1.75(m,1H),1.73-1.61(m,1H).

[0369] Characterization of compounds 1-3B: TLC developing solvent was petroleum ether:ethyl acetate = 1:1, R f =0.3; LCMS:456.1[M+H] + ; 1H NMR (400MHz, CDCl3) δ = 9.11 (s, 1H), 8.83 (br d,J=7.8Hz,1H),8.08(d,J=8.3Hz,2H),7.66(d,J=4.2Hz,1H),7.32(d,J=8.2Hz,2H),6.82(d,J=4.2Hz,1H),4.34-4.24(m,1H),4. 15-4.10(m,1H),3.78-3.61(m,1H),3.31(t,J=10.9Hz,1H),2.71-2.56(m,2H),2.41(s,3H),2.05-1.98(m,1H),1.80-1.63(m,2H).

[0370] Step 2: Under a nitrogen atmosphere, compound 1-3A (0.7 g, 1.54 mmol) was added to methanol (7 mL), followed by 5% palladium on carbon (0.2 g). After three purgings with hydrogen, the mixture was stirred at 25°C for 1 hour under a hydrogen atmosphere (15 psi). After the reaction was complete, the reaction solution was filtered, and the filtrate was concentrated under reduced pressure to obtain compound 1-4A. LCMS: 426.1 [M+H] + . 1 H NMR (400MHz, CDCl3) δ = 8.02 (d, J = 8.4Hz, 2H), 7.86 (s, 1H), 7.52 (d, J = 4.0Hz, 1H), 7.25 (d, J = 8.4Hz, 2H), 6.64 (d, J = 4.0Hz, 1H), 4.68 (br d,J=8.8Hz,1H),4.31-4.17(m,1H),3.91(br dd,J=2.8,4.0Hz,1H),3.68-3.59(m,1H),3.16(t,J=10.8Hz,1H),2.60(d,J=6.0Hz,2H),2.37(s,3H),2.34-2.27(m,1H),1.96(br dd,J=2.8,13.2Hz,2H),1.69-1.46(m,3H).

[0371] Step 3: Under a nitrogen atmosphere, compound 1-4A (600 mg, 1.41 mmol) was added to acetic acid (12 mL), followed by tetramethyl orthocarbonate (1.92 g, 14.1 mmol) and p-toluenesulfonic acid (24.28 mg, 0.14 mol). The reaction mixture was stirred at 25 °C for 8 hours. After the reaction was complete, 80 mL of saturated sodium bicarbonate was added to quench the reaction, and the mixture was extracted with ethyl acetate (80 mL × 3). The combined organic phases were 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 obtain compound 1-5A. LCMS: 466.1 [M+H] + . 1 H NMR (400MHz, CDCl3) δ = 8.65 (s, 1H), 8.09 (d, J = 8.4Hz, 2H), 7.79 (d, J = 4.0Hz, 1H), 7.25(s,2H),6.79(d,J=4.0Hz,1H),4.62(dtd,J=4.4,8.0,12.4Hz,1H),4.22(s,3H ),4.06(d,J=8.4Hz,2H),3.83(dtd,J=2.0,5.6,11.4Hz,1H),2.66(d,J=6.0Hz,2H) ,2.49(dq,J=4.4,12.8Hz,1H),2.36(s,3H),2.21-2.06(m,2H),1.84-1.68(m,1H).

[0372] Step 4: Under a nitrogen atmosphere, potassium hydroxide (280 mg) was dissolved in methanol (5 mL), and then compound 1-5A (200 mg, 430 μmol) was added. The reaction mixture was stirred at 25 °C for 4 hours. After the reaction was complete, the pH was adjusted to neutral with 4 M dilute hydrochloric acid solution, diluted with 60 mL of water, and extracted with ethyl acetate (60 mL × 3). The combined organic phases were washed with 120 mL of saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was purified by silica gel column chromatography (dichloromethane:methanol = 10:0-10:1) to obtain compound 1A. LCMS: 312.1 [M + H] + . 1H NMR(400MHz,DMSO-d6)δ=11.73(br s,1H),8.34(s,1H),7.45(t,J=2.8Hz,1H),6.71(dd,J=1.6,3.2Hz,1H),4.71-4.48(m,1H),4.13(s,3H),4.03(d,J=8.4Hz ,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 ID, 3μm; mobile phase: phase A is supercritical CO2, phase B is isopropanol (0.05% diethylamine); gradient (B%): 5%-40%), with a retention time of 1.700 min and a chiral purity of 96.74%.

[0373] Step 5: Following the above synthetic method, crude compound 1B was prepared using compounds 1-3B as raw materials. The crude product was then analyzed by prep-HPLC (column: Waters Xbridge 150*25mm*5μm; mobile phase: [water (NH3·H2O)-acetonitrile]; gradient (acetonitrile %): 12%-42%) to compound 1B. LCMS: 312.1 [M+H] + ; 1 H NMR(400MHz,DMSO-d6)δ=11.62(br s,1H),8.27(s,1H),7.36(t,J=3.2Hz,1H),6.61(dd,J=2.0,3.6Hz,1H),4.75-4.57(m,1H),4.07(s,3H),4.05-3.99(m,1H),3.84(dd,J =4.8,12.0Hz,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., 3μm; mobile phase: phase A is supercritical CO2, phase B is isopropanol (0.05% diethylamine); gradient (B%): 5%-40%), with a retention time of 1.773 min and a chiral purity of 99.08%.

[0374] Example 2

[0375]

[0376]

[0377] Step 1: Under a nitrogen atmosphere, compound 2-2 (23.8 g, 140 mmol) was added to tetrahydrofuran (300 mL), and the mixture was cooled to -78 °C. Butyllithium (2.5 M, 51.3 mL, 128 mmol) was added dropwise. After the addition was complete, the reaction mixture was allowed to react at -78 °C for 0.5 hours. Then, a tetrahydrofuran solution of compound 2-1 (30 g, 117 mmol) was added dropwise to the reaction mixture (100 mL). The mixture was stirred at -78 °C for 3 hours. The reaction was quenched by adding 100 mL of saturated ammonium chloride aqueous solution at 0 °C. The mixture was extracted with ethyl acetate (100 mL × 2). The combined organic phases were washed with 100 mL of saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain compound 2-3. 1 H NMR (400MHz, CDCl3) δ = 5.09 (br d,J=7.2Hz,1H),4.46(s,2H),4.32-4.25(m,1H),4.20(q,J=7.2Hz,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).

[0378] Step 2: Under a nitrogen atmosphere and at 0°C, compound 2-3 (49 g, 114 mmol) was added to DMF (400 mL), followed by the dropwise addition of hydrazine hydrate (8.7 g, 165 mmol). The reaction mixture was heated to 25°C and stirred for 2 hours. After the reaction was complete, the mixture was diluted with 100 mL of water and extracted with ethyl acetate (100 mL × 2). The combined organic phases were washed with 100 mL of saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain compound 2-4. 1 H NMR (400MHz, CDCl3) δ = 6.01 (s, 1H), 5.28 (br d,J=8.4Hz,1H),4.74(s,2H),4.40-4.28(m,1H),4.18(q,J=7.2Hz,2H),2.81-2.65(m,2H),2.19-2.07(m,1H),1.91(br dd,J=7.2,12.8Hz,1H),1.46(s,9H),1.26(t,J=7.1Hz,3H),0.92(s,9H),0.10(s,6H).

[0379] Step 3: Compound 2-4 (50 g, 113 mmol) was added to a mixed solution of tetrahydrofuran (400 mL) and methanol (60 mL) under a nitrogen atmosphere at 0 °C. Sodium borohydride (8.99 g, 238 mmol) was added in portions. The reaction mixture was heated to 25 °C and stirred for 12 hours. The reaction mixture was quenched with 50 mL of saturated ammonium chloride aqueous solution and extracted with ethyl acetate (100 mL × 2). The combined organic phases were washed with 100 mL of saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 10 / 1 to 0 / 1) to obtain compound 2-5. 1 H NMR (400MHz, CDCl3) δ = 5.99 (s, 1H), 5.02 (br d,J=6.8Hz,1H),4.73(s,2H),3.73-3.62(m,2H),3.61-3.55(m,1H),2.84-2.72(m,1 H), 2.70-2.54 (m, 1H), 1.84 (q, J = 6.8Hz, 2H), 1.46 (s, 9H), 0.92 (s, 9H), 0.10 (s, 6H).

[0380] Step 4: Under a nitrogen atmosphere at 0°C, compound 2-5 (22 g, 55.1 mmol) and 1,1-azodicarbonylpiperidine (23.3 g, 92.5 mmol) were added to tetrahydrofuran (200 mL), followed by the slow dropwise addition of tri-tert-butylphosphine (18.7 g, 92.5 mmol). After the addition was complete, the reaction mixture was heated to 25°C and stirred for 12 hours. The reaction mixture was quenched with 100 mL of saturated ammonium chloride aqueous solution, extracted with ethyl acetate (100 mL × 2), and the combined organic phases were washed with 100 mL of saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 20 / 1 to 1 / 1) to obtain compound 2-6. 1 H NMR (400MHz, CDCl3) δ = 6.02 (s, 1H), 4.72 (br d, J = 2.8Hz, 1H), 4.67 (s, 2H), 4.36-4.18 (m, 2H), 3.98-3.82 (m, 1H), 2.86 (br t,J=6.4Hz,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=0.7Hz,6H).

[0381] Step 5: Under a nitrogen atmosphere at 0°C, tetrabutylammonium fluoride (1M tetrahydrofuran solution, 14.4 mL, 14.4 mmol) was added to a tetrahydrofuran (50 mL) solution of compound 2-6 (5 g, 13.1 mmol). The reaction mixture was heated to 25°C and stirred for 1 hour. After the reaction was complete, the mixture was diluted with 50 mL of water, extracted with ethyl acetate (100 mL × 2), and the combined organic phases were 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.

[0382] Step 6: At room temperature, manganese dioxide (1.38 g, 131 mmol) was slowly added in portions to a mixture of compound 2-7 (3.5 g, 13.1 mmol) in dichloromethane (25 mL) and methanol (25 mL). The mixture was heated to 65 °C and stirred for 12 hours. The reaction solution was cooled to room temperature, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 20 / 1 to 1 / 1) to obtain compound 2-8. SFC analysis (column: Chiralpak AD-3 50 × 4.6 mm ID, 3 μm; mobile phase: A phase was supercritical CO2, B phase was ethanol (0.05% diethylamine); gradient (B%): 10%-60%) showed that compound 2-8 had a retention time of 0.939 min and a chiral purity of 98.87%. 1 H NMR (400MHz, CDCl3) δ = 9.92 (s, 1H), 6.55 (s, 1H), 4.71 (br s, 1H), 4.47 (dd, J = 4.8, 13.2Hz, 1H), 4.35-4.24 (m, 1H), 4.06 (br dd,J=6.8,12.8Hz,1H),2.94(br t,J=6.4Hz,2H),2.12(dtd,J=2.8,6.8,13.2Hz,1H),1.96(qd,J=6.6,13.6Hz,1H),1.47(s,9H).

[0383] Step 7: Under a nitrogen atmosphere, sodium acetate (232 mg, 2.83 mmol) and hydroxylamine hydrochloride (157 mg, 2.26 mmol) were added sequentially to a methanol (5 mL) solution of compound 2-8 (500 mg, 1.88 mmol). The reaction mixture was stirred at 25 °C for 0.5 hours. After the reaction was complete, the mixture was diluted with 10 mL of water and extracted with ethyl acetate (100 mL × 2). The combined organic phases were washed with 100 mL of saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain compounds 2-9. LCMS: 281.1 [M+1] + .

[0384] Step 8: Under a nitrogen atmosphere at 0°C, Burgess reagent (663 mg, 2.78 mmol) was slowly added to a dichloromethane (5 mL) solution of compound 2-9 (520 mg, 1.86 mmol). The reaction was heated to 25°C and stirred for 12 hours. After the reaction was complete, 5 mL of water was added to quench the reaction, and the mixture was extracted with ethyl acetate (100 mL × 2). The combined organic phases were washed with 100 mL of saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain compound 2-10. 1 H NMR (400MHz, CDCl3) δ = 6.42 (s, 1H), 4.69 (br d, J = 5.6Hz, 1H), 4.43 (dd, J = 4.8, 13.2Hz, 1H), 4.26 (br s,1H),4.02(dd,J=6.813.2Hz,1H),3.00-2.84(m,2H),2.11(dtd,J=2.8,6.8,13.2Hz,1H),1.96(td,J=7.2,14.0Hz,1H),1.46(s,9H).

[0385] Step 9: Under a nitrogen atmosphere and at 0°C, trimethyliodosilane (417 mg, 2.08 mmol) was slowly added to a dichloromethane (5 mL) solution of compound 2-10 (420 mg, 1.60 mmol), and the reaction was carried out at 0°C for 0.5 hours. After the reaction was complete, the reaction solution was concentrated under reduced pressure at room temperature to obtain crude hydroiodide of compound 2-11.

[0386] Step 10: Under a nitrogen atmosphere, compound 2-11 (438 mg, crude hydroiodide) and compound 2-12 (300 mg, 1.51 mmol) were added to an isopropanol (8 mL) solution, followed by N,N-diisopropylethylamine (1.37 g, 10.6 mmol). The reaction mixture was heated to 80 °C and stirred for 12 hours. After the reaction was complete, the reaction mixture was cooled to room temperature, diluted with 5 mL of water, and extracted with ethyl acetate (100 mL × 2). The combined organic phases were washed with 100 mL of saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 10 / 1 to 0 / 1) to obtain compound 2-13. SFC analysis (column: Chiralcel OJ-3 50×4.6mm ID, 3μm; mobile phase: A phase: supercritical CO2; B phase: ethanol (0.05% diethylamine); gradient (B%): 10%-60%) showed that the retention time of compound 2-13 was 2.087 min and the chiral purity was 100%. 1H NMR (400MHz, CDCl3) δ = 9.29 (s, 1H), 8.78 (br d,J=8.0Hz,1H),7.96(d,J=2.0Hz,1H),7.04(d,J=2.0Hz,1H),6.53(s,1H),5.23-5.09(m,1H),4.71(dd,J =4.8,13.2Hz,1H),4.28(dd,J=7.2,13.2Hz,1H),3.21-3.01(m,2H),2.49-2.37(m,1H),2.28-2.17(m,1H).

[0387] Step 11: Under a nitrogen atmosphere, 295 mg (5% palladium on carbon) was added to a mixed solution of 300 mg (925 μmol) of compound 2-13 in 4 mL of dichloromethane and 4 mL of methanol. After purging with hydrogen three times, the reaction mixture was stirred at 25 °C for 0.5 hours under a hydrogen atmosphere (15 psi). The reaction mixture was filtered, and the filtrate was concentrated under reduced pressure to obtain compound 2-14. LCMS: 295.1 [M+1] + .

[0388] Step 12: Under a nitrogen atmosphere, compound 2-14 (100 mg, 340 μmol) was added to a mixed solution of toluene (0.5 mL) and tetrahydrofuran (0.5 mL) at room temperature, followed by the sequential addition of trimethyl orthoacetate (81.7 mg, 680 μmol) and p-toluenesulfonic acid (5.85 mg, 34.0 μmol). The reaction mixture was heated to 100 °C and stirred for 1 hour. The reaction mixture was cooled to room temperature, diluted with 5 mL of water, and extracted with ethyl acetate (50 mL × 2). The combined organic phases were washed with 50 mL of saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was purified by silica gel column chromatography (dichloromethane / methanol = 1 / 0-10 / 1) to obtain compound 2. LCMS: 319.1 [M+1] + ; 1H NMR (400MHz, DMSO-d6) δ = 8.83 (s, 1H), 8.24 (d, J = 2.2Hz, 1H), 7.22 (d, J = 2.2Hz, 1H), 6.88 (s, 1H), 5.42-5.22 (m, 1H), 4 .85(dd,J=6.0,12.4Hz,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 analysis (Chiralcel OD-3 column 50*4.6mm ID, 3μm, mobile phase: A phase is supercritical CO2, B phase is isopropanol / acetonitrile (0.05% diethylamine); gradient (B%): 20%-60%) showed that the retention time of compound 2 was 1.073 min and the chiral purity was 99.89%.

[0389] Step 13: Following the steps above, compound 2A was prepared using compound 2-1A as the starting material. LCMS: 319.1 [M+1] + ; 1 HNMR (400MHz, DMSO-d6) δ=8.83(s,1H),8.24(d,J=2.4Hz,1H),7.23(d,J=2.4Hz,1H),6.88(s,1H),5.41-5.22(m,1H),4.85(dd,J =6.0,12.4Hz,1H),4.68-4.52(m,1H),3.25-3.17(m,1H),3.15-3.04(m,1H),2.74(s,3H),2.72-2.64(m,1H),2.38-2.29(m,1H). SFC detection (Chiralcel AD-3 50*4.6mm I.D., 3μm column; mobile phase: A phase is supercritical CO2, B phase is ethanol (0.05% diethylamine); gradient (B%): 10%-60%)) showed that the retention time of compound 2A was 2.150 min, and the chiral purity was 100%. The retention time of compound 2 detected by this method was 1.418 min.

[0390] Example 3

[0391]

[0392] Step 1: Under a nitrogen atmosphere, 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). The reaction mixture was heated to 70 °C and reacted for 13 hours. After cooling to room temperature, the reaction mixture was added to ice water (150 mL) and extracted with ethyl acetate (100 mL × 3). The combined organic phases were washed successively with water (50 mL × 3) and saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product. This crude product was purified by silica gel column chromatography (petroleum ether:ethyl acetate = 1:0-3:1) to obtain compound 3-3. LCMS: 183.1 [M + H - 56] + .

[0393] Step 2: Under a nitrogen atmosphere, palladium on carbon (2 g, 5% purity) was added to a methanol (100 mL) solution of compound 3-3 (10 g, 42.0 mmol). After purging with hydrogen three times, the mixture was stirred at 25 °C for 15 hours under a hydrogen atmosphere (15 psi). The reaction solution was filtered, and the filtrate was concentrated under reduced pressure. A mixture of ethyl acetate / petroleum ether (V / V = 5 / 1, 30 mL) was added to the residue, and the mixture was stirred at room temperature for 30 minutes. The mixture was filtered, and the filter cake was collected and dried under vacuum to obtain compound 3-4. 1 ¹H NMR (400MHz, CDCl₃) δ=4.30(s,1H),4.16-4.02(m,1H),3.74-3.40(m,2H),3.04(t,J=10.8Hz,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 spectroscopy confirmed the structure of compounds 3-4.

[0394] Step 3: Under a nitrogen atmosphere at 0°C, trimethyliodosilane (3.30 g, 16.5 mmol) was added to a dichloromethane (40 mL) solution of compound 3-4 (3.6 g, 15.0 mmol), and the reaction solution was reacted at 0°C for 1 hour. The reaction solution was concentrated under reduced pressure to obtain crude compound 3-5.

[0395] Step 4: Under a nitrogen atmosphere, N,N-diisopropylethylamine (4.20 g, 32.5 mmol) was added 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). The reaction solution was heated to 90 °C and reacted for 2 hours. The reaction solution was cooled to room temperature, concentrated, and the residue was dissolved in ethyl acetate (100 mL). The residue was washed successively with water (20 mL) and saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was purified by silica gel column chromatography (petroleum ether:ethyl acetate = 10:1-1:1) to obtain compound 3-6. LCMS: 456.0 [M+H] + . 1 H NMR (400MHz, CDCl3) δ = 9.11 (s, 1H), 8.83 (d, J = 8.0Hz, 1H), 8.07 (d, J = 8.4Hz, 2H),7.66(d,J=4.0Hz,1H),7.32(d,J=8.0Hz,2H),6.82(d,J=4.4Hz,1H),4.3 6-4.25(m,1H),4.16-4.06(m,1H),3.79-3.64(m,1H),3.31(t,J=11.0Hz,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).

[0396] Step 5: Under a nitrogen atmosphere, palladium on carbon (1 g, 5% concentration) was added to a methanol (50 mL) solution of compounds 3-6 (4.65 g, 10.2 mmol). After purging with hydrogen three times, the mixture was stirred for 15 hours under a hydrogen atmosphere (15 psi) at 25 °C. The reaction solution was filtered, and the filtrate was concentrated under reduced pressure to obtain compounds 3-7. LCMS: 426.0 [M+H] + ; 1H NMR (400MHz, CDCl3) δ = 8.02 (d, J = 8.0Hz, 2H), 7.85 (s, 1H), 7.51 (d, J = 4.0Hz, 1H), 7.25 (d, J=8.0Hz,2H),6.63(d,J=4.0Hz,1H),4.65(d,J=8.4Hz,1H),4.35-4.13(m,1H),3.97-3.78( m,1H),3.70-3.56(m,1H),3.16(t,J=10.8Hz,1H),2.59(d,J=5.6Hz,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).

[0397] Step 6: Under a nitrogen atmosphere, trimethyl orthoacetate (451.79 mg, 3.76 mmol) and p-toluenesulfonic acid (32.38 mg, 188.01 μmol) were added to a toluene / tetrahydrofuran (V / V = 1 / 1, 16 mL) mixed solution of compounds 3-7 (800 mg, 1.88 mmol). The reaction solution was stirred at 100 °C for 1 hour. The reaction solution was cooled to room temperature, diluted with water (50 mL), and extracted with ethyl acetate (50 mL × 3). The combined organic phases were washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product. This crude product was then purified by silica gel column chromatography (petroleum ether:ethyl acetate = 1:1-0:1) to obtain compounds 3-8. LCMS: 450.1 [M+H] + ; 1 H NMR (400MHz, chloroform-d) δ = 8.81 (s, 1H), 8.11 (d, J = 8.4Hz, 2H), 7.84 (d, J = 4.0Hz, 1H), 7.28 (s, 1H), 7.26 (s, 1H), 6.88 (d, J = 4.0Hz, 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).

[0398] Step 7: Under a nitrogen atmosphere, add 2M potassium hydroxide aqueous solution (8mL) to a methanol (800mg, 1.78mmol) solution of compounds 3-8. Heat the reaction solution to 65°C and stir for 2 hours. Cool the reaction solution to room temperature, dilute with water (50mL), extract with ethyl acetate (50mL × 3), wash the combined organic phases with saturated brine (100mL), dry to anhydrous sodium sulfate, filter, concentrate the filtrate under reduced pressure to obtain the crude product, and then purify by silica gel column chromatography (dichloromethane:methanol = 1:0-10:1) to obtain compound 3. LCMS: 296.1 [M + H] + ; 1 H NMR (400MHz, DMSO-d6) δ = 11.85 (br s, 1H), 8.47 (s, 1H), 7.47 (t, J = 3.2Hz, 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=13.1Hz,1H),1.83-1.63(m,1H). SFC analysis (analytical method: column: Chiralcel OJ-3 50*4.6mm ID, 3μm; mobile phase: phase A is supercritical CO2, phase B is isopropanol (0.05% diethylamine); gradient (B%): 5%-40%) showed that compound 3 had a retention time of 1.513 min and a chiral purity of 100%.

[0399] Example 4

[0400]

[0401]

[0402] Step 1-2: Using compounds 4-1 and 1-2 as raw materials, compound 4-3 was prepared according to the method in Example 1.

[0403] Step 3: Under a nitrogen atmosphere, N,N-thiocarbonyldiimidazole (266.27 mg, 1.49 mmol) was added to a toluene (10 mL) solution of compound 4-3 (300 mg, 747.06 μmol). The reaction solution was heated to 80 °C and stirred for 1 hour. After cooling to room temperature, the solution was diluted with ethyl acetate (120 mL), washed successively with water (100 mL × 3) and saturated brine (120 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product. This crude product was then purified by silica gel column chromatography (petroleum ether:ethyl acetate = 2:1-1:2) to obtain compound 4-4. LCMS: 444.1 [M+H] + ; 1 H NMR (400MHz, CDCl3) δ = 10.54 (br s, 1H), 8.35 (br s,1H),7.52(d,J=3.6Hz,1H),6.82(d,J=3.6Hz,1H),5.76(s,2H),4.48-3.94(m,3H),3.64-3.42(m,2H),2.68(br d,J=5.6Hz,2H),2.18(br d,J=12.0Hz,2H),1.97-1.67(m,3H),0.99-0.85(m,2H),-0.05(s,9H).

[0404] Step 4: Under a nitrogen atmosphere, iodomethane (143.98 mg, 1.01 mmol) was added to a solution of compound 4-4 (300.00 mg, 676.23 μmol) and potassium carbonate (140.19 mg, 1.01 mmol) in N,N-dimethylformamide (5 mL). The reaction mixture was stirred at 25 °C for 1 hour. Ethyl acetate (80 mL) was added to the reaction mixture, followed by washing with water (70 mL × 3) and saturated brine (100 mL) successively. The mixture was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 1:1-1:2) to obtain compound 4-5. LCMS: 458.1 [M+H] + .

[0405] Step 5: Add m-chloroperoxybenzoic acid (267 mg, 1.32 mmol, 85% purity) in portions to a solution of compounds 4-5 (250 mg, 546.25 μmol) in dichloromethane (5 mL). After the addition is complete, stir the reaction mixture at 25 °C for 1 hour. Add 80 mL of dichloromethane to the reaction mixture, wash successively with water (70 mL × 3) and saturated brine (100 mL), dry to anhydrous sodium sulfate, filter, concentrate the filtrate under reduced pressure to obtain the crude product, and purify by silica gel column chromatography (petroleum ether: ethyl acetate = 2:1-1:2) to obtain compounds 4-6. LCMS: 490.2 [M + H] + ; 1H NMR (400MHz, CDCl3) δ = 8.87 (s, 1H), 7.53 (d, J = 3.6Hz, 1H), 6.88 (d, J = 3.6Hz, 1H), 5.82 (s, 2H),5.50-5.27(m,1H),4.48-4.34(m,1H),4.24(dd,J=4.4,10.4Hz,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=6.4,9.6Hz,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).

[0406] Step 6: Under a nitrogen atmosphere at 0°C, sodium hydride (36.76 mg, 919.00 μmol, 60% purity) was added to a tetrahydrofuran (2 mL) solution of cyclopropanol (53.37 mg, 919.00 μmol). The reaction mixture was stirred at 0°C for 0.5 hours. Then, at 0°C, a tetrahydrofuran (2 mL) solution of compound 4-6 (150 mg, 306.33 μmol) was added dropwise to the reaction mixture. After the addition was complete, the reaction mixture was heated to 25°C and reacted for 1 hour. The reaction mixture was cooled to 0°C, quenched with 40 mL of water, and extracted with ethyl acetate (40 mL × 3). The combined organic phases were washed with 80 mL of saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 2:1-0:1) to obtain compound 4-7. LCMS: 468.3 [M+H] + .

[0407] Step 7: Under a nitrogen atmosphere, a solution of compound 4-7 (50 mg, 106.92 μmol) in 1 mL of trifluoroacetic acid was stirred at 25 °C for 1 hour. The reaction solution was concentrated under reduced pressure to obtain crude compound 4-8. LCMS: 368.1 [M+H] + .

[0408] Step 8: Under a nitrogen atmosphere, compounds 4-8 (35 mg, crude product) were added to ammonia water (2 mL) and stirred at 25 °C for 1 hour. The reaction solution was concentrated under reduced pressure to obtain the crude product, which was then preparatively separated by reversed-phase column chromatography (column: Phenomenex Luna C18 150*25 mm*10 μm; mobile phase: H2O (0.225% formic acid)-acetonitrile; gradient (acetonitrile%): 10%-40%) to obtain compound 4. LCMS: 338.1 [M+H] + ; 1H NMR(400MHz,DMSO-d6)δ=11.76(br s,1H),8.38(s,1H),7.46(t,J=3.0Hz,1H),6.70(dd,J=1.7,3.2Hz,1H),4.63-4.43(m,2H),4.05-3.91(m,2 H),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 (column: Chiralcel OJ-3 50*4.6mm ID, 3μm; mobile phase: A phase is supercritical CO2, B phase is isopropanol (0.05% diethylamine); gradient (B%): 5%-40%), the retention time of compound 4 was 1.376 min, and the chiral purity was 100%.

[0409] Example 5

[0410]

[0411] Step 1: Under a nitrogen atmosphere, ferric trifluoromethanesulfonate (11.27 mg, 22.41 μmol) and compound 5-1 (55.96 mg, 268.94 μmol) were added to a solution of compound 4-3 (90 mg, 224.12 μmol) in N,N-dimethylformamide (1.5 mL). The reaction solution was heated to 60 °C and stirred for 0.5 hours. The reaction solution was cooled to room temperature and diluted with 60 mL of ethyl acetate. The organic phase was washed successively with water (60 mL × 3) and saturated brine (60 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product. This crude product was purified by silica gel column chromatography (petroleum ether:ethyl acetate = 2:1-1:2) to obtain compound 5-2. LCMS: 480.2 [M+H] + , 1HNMR (400MHz, CDCl3) δ = 8.94 (s, 1H), 7.53 (d, J = 3.6Hz, 1H), 6.87 (d, J = 3.6Hz, 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=2.0,5.9Hz ,2H),2.34-2.14(m,2H),1.95-1.81(m,1H),0.99-0.89(m,2H),-0.05(s,9H).

[0412] Step 2: Under a nitrogen atmosphere, a solution of compound 5-2 (90 mg, 187.67 μmol) in 2.5 mL of trifluoroacetic acid was stirred at 25 °C for 1 hour. The reaction solution was concentrated under reduced pressure to obtain crude compound 5-3. LCMS: 380.1 [M+H] + .

[0413] Step 3: Under a nitrogen atmosphere, a solution of compound 5-3 (70 mg, 184.53 μmol) in ammonia water (2.5 mL) was stirred at 25 °C for 1 hour. The reaction solution was concentrated under reduced pressure to obtain a crude product, which was then preparatively separated by reversed-phase column chromatography (Phenomenex Luna C18 150*25 mm*10 μm column; mobile phase: H2O (0.225% formic acid)-acetonitrile; gradient (acetonitrile%): 30%-60%) to obtain compound 5. LCMS: 350.1 [M+H] + ; 1 HNMR (400MHz, DMSO-d6) δ = 12.32 (br s, 1H), 8.78 (s, 1H), 7.64 (t, J = 3.2Hz, 1H), 6.98 (br d,J=1.6Hz,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=12.6Hz,1H),2.08(br d,J=13.7Hz,1H),1.83-1.62(m,1H); 19F NMR (376MHz, DMSO-d6) δ=-59.898; SFC detection (Chiralpak IC-3 column 50×4.6mm I.D., 3μm; 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, and the chiral purity was 100%.

[0414] Example 6

[0415]

[0416] Step 1: Under a nitrogen atmosphere, 1,1-carbonyldiimidazole (969 mg, 5.98 mmol) was added to a toluene (15 mL) solution of compound 4-3 (1.2 g, 2.99 mmol). The reaction mixture was heated to 60 °C and stirred for 2 hours. The reaction mixture was cooled to room temperature, diluted with 120 mL of ethyl acetate, washed successively with water (100 mL × 3) and saturated brine (120 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product. This crude product was purified by silica gel column chromatography (petroleum ether:ethyl acetate = 5:1-2:1) to obtain compound 6-1. LCMS: 428.1 [M+H] + ; 1 H NMR (400MHz, CDCl3) δ = 8.16 (s, 1H), 7.42 (d, J = 3.6Hz, 1H), 6.69 (d, J = 3.6Hz, 1H), 5.71 (s, 2H), 4.78-4.60 (m, 1H), 4.40 (br t,J=11.2Hz,1H),4.21-4.09(m,1H),3.96-3.83(m,1H),3.65-3.51(m,2H),2. 83(dq,J=4.0,12.8Hz,1H),2.67(d,J=5.6Hz,2H),2.29-2.17(m,1H),2.11(br d,J=12.4Hz,1H),1.86-1.77(m,1H),1.00-0.86(m,2H),-0.05(s,9H).

[0417] Step 2: Under a nitrogen atmosphere at 0°C, sodium hydroxide (171 mg, 4.29 mmol, 60% purity) was added to a solution of compound 6-1 (600 mg, 1.40 mmol) in N,N-dimethylformamide (7 mL). After the addition was complete, the reaction mixture was stirred at 0°C for 0.5 hours. Then, CF2Br2 (1.96 g, 7.02 mmol) was added to the reaction mixture, and stirring was continued at 0°C for 2 hours. The reaction solution was quenched with 30 mL of ammonium chloride aqueous solution, extracted with ethyl acetate (60 mL × 3), and the combined organic phases were washed with 50 mL of saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 10:1-0:1), followed by SFC separation (column: DAICELCHIRALCEL OD (250 mm × 30 mm, 10 μm); mobile phase: A phase was supercritical CO2, B phase was ethanol (0.1% NH3H2O); gradient (B%): 25%) to obtain compound 6-2. LCMS: 556.0, 558.1 [M+H] + ; 1 H NMR (400MHz, CDCl3) δ = 8.84 (s, 1H), 7.56-7.51 (m, 1H), 6.82 (d, J = 3.6Hz, 1H), 5.84 (s, 2H), 4.98-4.76 (m, 1H), 4.32-4.10 (m, 2H), 3.97(td,J=5.2,10.4Hz,1H),3.67-3.61(m,2H),2.76(d,J=5.6Hz,2H),2.62(dq,J=4.0,12.8Hz,1H),2.46-2.34(m,1H),2.23(br d, J=14.4Hz, 1H), 1.91 (dq, J=4.0, 12.8Hz, 1H), 1.03-0.97 (m, 2H), 0.01 (s, 9H).

[0418] Step 3: Under a nitrogen atmosphere at room temperature, silver tetrafluoroborate (153 mg, 784 μmol) was added in portions to a 1 mL solution of compound 6-2 (58 mg, 104 μmol) in dichloromethane. The reaction mixture was reacted at 25 °C for 22 hours. The reaction mixture was filtered, and the filtrate was purified by reversed-phase column chromatography (Phenomenex luna C18 column (250 × 70 mm, 10 μm); mobile phase: water (0.1% formic acid)-acetonitrile; gradient (acetonitrile %): 55%-100%) to obtain compound 6-3. LCMS: 496.2 [M+H] + .

[0419] Step 4: Under a nitrogen atmosphere, trifluoroacetic acid (0.15 mL) was added to a solution of compound 5 (10 mg, 20.18 μmol) in dichloromethane (0.3 mL). The reaction mixture was stirred at 25 °C for 12 hours. The reaction mixture was concentrated under reduced pressure to obtain crude compounds 6-4. LCMS: 396.1 [M+H] + .

[0420] Step 5: Under a nitrogen atmosphere, ammonia (20.7 μL, 30% concentration) was added to a tetrahydrofuran (0.3 mL) solution of compound 6-4 (7 mg, crude product). The reaction mixture was stirred at 25 °C for 8 hours. The reaction mixture was filtered, and the filtrate was purified by reversed-phase column chromatography (column: Phenomenexluna C18 150 × 25 mm × 10 μm; mobile phase: H2O (0.225% formic acid) - acetonitrile; gradient (acetonitrile %): 20%-50%) to obtain compound 6. LCMS: 366.1 [M+H] + ; 1 H NMR(400MHz, CDCl3)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=5.2Hz,2H),2.61-2.51(m,1H),2.38-2.31(m,1H),2.19(br d,J=13.6Hz,1H),1.90(s,1H); 19 F NMR (376MHz, DMSO-d6) δ = -57.80.

[0421] Example 7

[0422]

[0423] Step 1: Under a nitrogen atmosphere, 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), and the reaction was carried out at 70 °C for 12 hours. The reaction solution was cooled to room temperature, 200 mL of water was added, and the mixture was extracted with ethyl acetate (200 mL × 2). The combined organic phases were washed with 200 mL of saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product. Compound 7-2 was then purified by silica gel column chromatography (petroleum ether:ethyl acetate = 10:1–3:1) to obtain compound 7-2. 1H NMR (400MHz, DMSO-d6) δ = 7.11 (br d, J = 5.8Hz, 1H), 4.94-4.64 (m, 1H), 4.11 (br d, J = 6.5Hz, 1H), 3.87-3.55 (m, 2H), 2.79-2.55 (m, 1H), 1.95 (br d,J=6.5Hz,1H),1.78-1.50(m,1H),1.39(d,J=1.9Hz,9H).

[0424] Step 2: Under a nitrogen atmosphere, palladium on carbon (2.01 g, 5% concentration) was added to a methanol (50 mL) solution of compound 7-2 (4.5 g, 18.89 mmol). After purging with hydrogen three times, the reaction was carried out under a hydrogen atmosphere (15 Psi) at 25 °C for 5 hours. The reaction solution was filtered, and the filtrate was concentrated under reduced pressure to obtain compound 7-3. 1 H NMR (400MHz, 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=8.9Hz,9H),1.36-1.21(m,1H).

[0425] Step 3: Under a nitrogen atmosphere at 0°C, trimethyliodosilane (5.05 g, 25.25 mmol) was added to a dichloromethane (50 mL) solution of compound 7-3 (5.02 g, 20.91 mmol). The reaction mixture was stirred at 0°C for 1 hour. The reaction mixture was filtered, and the filtrate was concentrated under reduced pressure to obtain crude hydroiodide of compound 7-4.

[0426] Step 4: Under a nitrogen atmosphere, compound 1-1 (4.07 g, 11.56 mmol) and N,N-diisopropylethylamine (7.47 g, 57.82 mmol) were added to a tert-butanol (30 mL) solution of compound 7-4 (3.1 g, crude hydroiodide). The reaction solution was heated to 90 °C and stirred for 2.5 hours. The reaction solution was cooled to room temperature, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 10:1-1:1) to obtain compound 7-5A (petroleum ether: ethyl acetate = 1:1, R... f =0.45) and compound 7-5B (petroleum ether: ethyl acetate = 1:1, R f =0.4).

[0427] Characterization of compound 7-5A: 1H NMR (400MHz, DMSO-d6) δ=8.89(s,1H),8.62(d,J=8.4Hz,1H),8.00(d,J=8.4Hz,2H),7.85(d,J=4.0Hz,1H),7.44(d,J=8.4H z,2H),6.96(d,J=4.4Hz,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=12.0Hz,1H),1.80(br d,J=3.2Hz,2H),1.66-1.48(m,1H).

[0428] Characterization of compound 7-5B: 1 H NMR (400MHz, DMSO-d6) δ = 9.40 (d, J = 8.0Hz, 1H), 8.93 (s, 1H), 8.00 (d, J = 8.4Hz, 2 H),7.80(d,J=4.0Hz,1H),7.44(d,J=8.4Hz,2H),7.02(d,J=4.4Hz,1H),4.35(br d,J=7.6Hz,1H),3.98(br d,J=12.6Hz,1H),3.79(d,J=10.8Hz,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=1.6Hz,1H),1.52-1.40(m,1H).

[0429] Step 5: Under a nitrogen atmosphere, palladium on carbon (1.31 g, 5% purity) was added to a methanol (15 mL) solution of compound 7-5A (1.4 g, 3.07 mmol). After purging with hydrogen three times, the mixture was stirred for 2 hours under a hydrogen atmosphere (15 Psi) at 25 °C. The reaction solution was filtered, and the filtrate was concentrated under reduced pressure to obtain compound 7-6A. 1H NMR (400MHz, DMSO-d6) δ = 7.90 (d, J = 8.4Hz, 2H), 7.58 (s, 1H), 7.53 (d, J = 4.4Hz, 1H), 7.38 (d, J = 8.4Hz, 2H), 6.69 (d, J = 4.4Hz, 1H), 5.09 (d, J=8.8Hz,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=12.0Hz,1H),1.88-1.76(m,1H),1.66-1.43(m,2H).

[0430] Step 6: Under a nitrogen atmosphere, p-toluenesulfonic acid monohydrate (6.71 mg, 35.25 μmol) was added to a 2 mL acetic acid solution containing tetramethyl carbonate (479.95 mg, 3.53 mmol) and compound 7-6A (150 mg, 352.52 μmol). The reaction solution was reacted at 25 °C for 12 hours. The reaction solution was concentrated to obtain a crude product, which was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 100:1-1:2) to obtain compound 7-7A. LCMS: 466.1 [M+1] + .

[0431] Step 7: Under a nitrogen atmosphere, potassium hydroxide aqueous solution (2 mL, 1 M) was added to a methanol (4 mL) solution of compound 7-7A (110 mg, 236.29 μmol). The reaction solution was heated to 60 °C and reacted for 4 hours. The pH of the reaction solution was adjusted to 7-7.5 with 1 M dilute hydrochloric acid, and the crude product was concentrated under reduced pressure. Compound 7A was then preparatively separated by reversed-phase column chromatography (Waters Xbridge C18 150*25 mm*5 μm column; mobile phase: H2O (0.05% NH3H2O)-acetonitrile; gradient (acetonitrile%): 6%-36%). LCMS: 312.2 [M+1] + ; 1¹H NMR (400MHz, CDCl₃) δ=10.59(br s, 1H), 8.60(s, 1H), 7.42(d, J=2.0Hz, 1H), 6.71(d, J=3.2Hz, 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, 11.2Hz, 1H), 2.69(d, J=6.0Hz, 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 (Chiralcel OD-3 column 50*4.6mm) ID, 3 μm; Mobile phase: Phase A is supercritical CO2, Phase B is isopropanol (0.05% diethylamine); Gradient (B%): 5%-40%), the retention time of compound 7A is 1.636 min, and the chiral purity is 98.77%.

[0432] Step 8: Referring to steps 5-7 above, compound 7B was prepared using compound 7-5B as the starting material. LCMS: 312.2 [M+1] + .

[0433] Example 8

[0434]

[0435] Step 1: Under a nitrogen atmosphere, p-toluenesulfonic acid monohydrate (6.07 mg, 35.25 μmol) was added to a mixed solution of trimethyl orthoacetate (84.71 mg, 705.05 μmol) and compound 7-6A (150 mg, 352.52 μmol) in toluene (1 mL) and tetrahydrofuran (1 mL). The reaction solution was heated to 100 °C and stirred for 3 hours. The reaction solution was cooled to room temperature and concentrated under reduced pressure to obtain a crude product, which was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 10:1-0:1) to obtain compound 8-1A. LCMS: 450.1 [M+1] + .

[0436] Step 2: Under a nitrogen atmosphere, potassium hydroxide aqueous solution (1.20 mL, 1.0 M) was added to a methanol (1 mL) solution of compound 8-1A (60 mg, 133.47 μmol). The reaction solution was stirred at 25 °C for 12 hours. The pH of the reaction solution was adjusted to 7-7.5 with 1 M dilute hydrochloric acid, and the crude product was concentrated under reduced pressure. Compound 8A was then preparatively separated by reversed-phase column chromatography (Waters Xbridge C18 150*25 mm*5 μm column; mobile phase: H2O (0.05% NH3·H2O)-acetonitrile; gradient (B%): 5%-35%). LCMS: 296.2 [M+1]+ ; 1 H NMR(400MHz,DMSO-d6)δ=11.89(br s,1H),8.49(s,1H),7.50(d,J=3.2Hz,1H),6.82(d,J=3.2Hz,1H),4.71-4.55(m,1 H),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=12.0Hz, 1H), 2.04 (br d,J=13.6Hz, 1H), 1.75 (dq, J=4.0, 12.8Hz, 1H). SFC analysis (Chiralpak AD-3 50*4.6mm ID, 3μm column; mobile phase: A phase is supercritical CO2, B phase is isopropanol + acetonitrile (0.05% diethylamine); gradient (B%): 20%-60%) showed that the retention time of compound 8A was 1.176 min and the chiral purity was 98.83%.

[0437] Step 3: Referring to Step 1, compound 8-1B was prepared using compound 7-6B as the starting material. Under a nitrogen atmosphere, magnesium powder (21.09 mg, 867.58 μmol) was added to a methanol (1 mL) solution of compound 8-1B (20 mg, 43.38 μmol), and the reaction solution was stirred at 25 °C for 2 hours. The reaction solution was filtered, and the filtrate was concentrated under reduced pressure to obtain a crude product. Compound 8B was then preparatively separated by reversed-phase chromatography (column: Waters Xbridge C18 150*25 mm*5 μm; mobile phase: H2O (0.05% NH3H2O)-acetonitrile; gradient (acetonitrile %): 5%-35%). LCMS: 296.1 [M+1] + ; 1H NMR(400MHz,DMSO-d6)δ=11.80(br s,1H),8.46(s,1H),7.46(d,J=3.2Hz,1H),6.73(d,J=3.2Hz,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 analysis (Chiralpak AD-350*4.6mm ID, 3μm column; mobile phase: A phase is supercritical CO2, B phase is isopropanol + acetonitrile (0.05% diethylamine); gradient (B%): 20%-60%) showed that the retention time of compound 8B was 1.422 min and the chiral purity was 99.54%.

[0438] Example 9

[0439]

[0440] Following the synthesis method described in the above embodiments, crude compound 9A was prepared and then separated by reversed-phase column chromatography (Phenomenex Luna C18 column 150*25mm*10μm; mobile phase: H2O (0.225% formic acid)-acetonitrile; gradient (acetonitrile%): 30%-50%) to obtain 9A. LCMS: 350.1 [M+1] + ; 1 H NMR (400MHz, DMSO-d6) δ = 12.32 (br s, 1H), 8.79 (s, 1H), 7.65 (t, J = 3.0Hz, 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(brd,J=11.5Hz,1H),2.08(br d,J=13.6Hz,1H),1.81-1.61(m,1H); 19 F NMR (376MHz, DMSO-d6) δ=59.91. SFC detection (Chiralpak AD-3 column 50×4.6mm ID, 3μm; mobile phase: A phase is supercritical CO2, B phase is ethanol (0.05% diethylamine); gradient (B%): 10%-60%)) The retention time of compound 9A was 0.624 min, and the chiral purity was 100%.

[0441] Example 10

[0442]

[0443] Step 1: Referring to the above examples, compound 2-1A was used as a raw material to prepare compound 2-7A.

[0444] Step 2: Under a nitrogen atmosphere at 0°C, methanesulfonyl chloride (790 μL, 10.2 mmol) was added to a solution of compound 2-7A (1 g, 3.74 mmol) and triethylamine (2.08 mL, 15.0 mmol) in dichloromethane (10 mL). The reaction mixture was heated to 25°C and stirred for 1 hour. 50 mL of water was added to the reaction mixture, and the mixture was extracted with ethyl acetate (100 mL × 2). The combined organic phases were washed with 100 mL of saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain compound 10-2.

[0445] Step 3: Under a nitrogen atmosphere, sodium cyanide (810 mg, 16.5 mmol) was added to a solution of compound 10⁻² (1.29 g, 3.73 mmol) in N,N-dimethylacetamide (10 mL). The reaction mixture was stirred at 25 °C for 1 hour. 50 mL of water was added to the reaction mixture, and the mixture was extracted with ethyl acetate (100 mL × 2). The combined organic phases were washed with 100 mL of saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 10 / 1 to 0 / 1) to obtain compound 10⁻³. LCMS: 221.1 [M⁻⁵⁶ + H] + .

[0446] Step 4: Under a nitrogen atmosphere at 0°C, trimethyliodosilane (59.1 μL, 434 μmol) was slowly added to a solution of compound 10⁻³ (100 mg, 362 μmol) in dichloromethane (1 mL). The reaction mixture was stirred at 0°C for 0.5 hours. The reaction mixture was then concentrated under reduced pressure to obtain crude hydroiodide of compound 10⁻⁴.

[0447] Step 5: Under a nitrogen atmosphere, N,N-diisopropylethylamine (286 μL, 1.64 mmol) was added to a solution of compound 10-4 (100 mg, crude hydroiodate) and compound 2-12 (78.4 mg, 395 μmol) in isopropanol (2 mL). The reaction mixture was heated to 80 °C and stirred for 12 hours. The reaction mixture was cooled to room temperature, diluted with 5 mL of water, and extracted with ethyl acetate (100 mL × 2). The combined organic phases were washed with 100 mL of saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 10 / 1 to 0 / 1) to obtain compound 10-5.1 H NMR (400MHz, DMSO-d6) δ = 9.08 (s, 1H), 8.62 (d, J = 8.8Hz, 1H), 8.45 (d, J = 2.0Hz, 1H), 7.17 (d, J = 2.0Hz, 1H), 6.07 (s, 1H), 5.14 -4.95(m,1H),4.50(dd,J=4.8,12.4Hz,1H),4.23(dd,J=8.0,12.4Hz,1H),3.92(s,2H),2.99-2.87(m,2H),2.31-2.09(m,2H). SFC analysis (Chiralcel OJ-3 column 50*4.6mm I.D., 3μm, mobile phase: A phase is supercritical CO2, B phase is isopropanol (0.05% diethylamine); gradient (B%): 5%-40%)) showed that the retention time of compound 10⁻⁵ was 1.357 min and the chiral purity was 99.43%.

[0448] Step 6: Under a nitrogen atmosphere, palladium on carbon (94.4 mg, 5% purity) was added to a mixed solution of compound 10⁻⁵ (100 mg, 295 μmol) in dichloromethane (2 mL) and methanol (2 mL). Hydrogen was then purged three times, and the mixture was stirred for 0.5 hours at 25°C under a hydrogen atmosphere (15 psi). The reaction solution was filtered, and the filtrate was concentrated under reduced pressure to obtain compound 10⁻⁶. LCMS: 309.1 [M+1] + .

[0449] Step 7: Under a nitrogen atmosphere at room temperature, trimethyl orthoacetate (70.1 mg, 584 μmol) and p-toluenesulfonic acid (5.03 mg, 29.2 μmol) were added sequentially to a mixed solution of compound 10-6 (90 mg, 292 μmol) in toluene (1 mL) and tetrahydrofuran (1 mL). The reaction mixture was heated to 70 °C and stirred for 5 hours. The reaction mixture was cooled to room temperature, diluted with 5 mL of water, and extracted with ethyl acetate (50 mL × 2). The combined organic phases were washed with 50 mL of saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was purified by silica gel column chromatography (dichloromethane / methanol = 1 / 0-10 / 1) to obtain compound 10. LCMS: 333.1 [M+1] + ; 1HNMR (400MHz, DMSO-d6) δ = 8.83 (s, 1H), 8.27 (d, J = 2.0Hz, 1H), 7.23 (d, J = 2.0Hz, 1H), 6.14 (s, 1H), 5.31-5.13 (m, 1H), 4.69 (d d,J=6.0,12.0Hz,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 analysis (Chiralcel OJ-3 column 50*4.6mm ID, 3μm, mobile phase: A phase is supercritical CO2, B phase is isopropanol (0.05% diethylamine); gradient (B%): 5%-40%) showed that the retention time of compound 10 was 1.326 min and the chiral purity was 100%.

[0450] Example 11

[0451]

[0452] Step 1: Under a nitrogen atmosphere, compounds 11-1 (9.20 g, 47.18 mmol) and 11-2 (6.56 g, 47.18 mmol) were stirred in 100 mL of ethylene glycol dimethyl ether at 25 °C for 1 hour. The reaction solution was concentrated under reduced pressure to remove the ethylene glycol dimethyl ether. The residue was dissolved in 100 mL of ethanol, and then heated to 80 °C and stirred for 8 hours. The reaction solution was cooled to room temperature and concentrated under reduced pressure to obtain 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 (400MHz, DMSO-d6) δ = 8.83 (s, 1H), 8.76 (dd, J = 0.8, 7.6Hz, 1H), 8.67 (d, J = 2.4 Hz, 1H), 7.76 (dd, J = 2.4, 7.6Hz, 1H), 4.35 (q, J = 7.2Hz, 2H), 1.34 (t, J = 7.2Hz, 3H).

[0453] Step 2: Under a nitrogen atmosphere, platinum dioxide (290 mg, 1.28 mmol) was added to a solution of compound 11-3 (3.0 g, 12.76 mmol) in ethanol (200 mL) and hydrochloric acid aqueous solution (1 M, 20 mL). The mixture was purged with hydrogen three times, and then stirred for 12 hours under a hydrogen atmosphere (50 psi) at 50 °C. The reaction solution was cooled to room temperature, filtered, and the filtrate was concentrated under reduced pressure to obtain crude hydrochloride of compound 11-4. LCMS: 210.1 [M+1] + .

[0454] Step 3: Under a nitrogen atmosphere, triethylamine (3.48 g, 34.4 mmol) and di-tert-butyl dicarbonate (Boc anhydride, 3.75 g, 17.2 mmol) were added to a dichloromethane (50 mL) solution of compound 11-4 (2.4 g, crude hydrochloride). The mixture was stirred at 25 °C for 2 hours. Water (80 mL) was added to the reaction solution, and the mixture was extracted with dichloromethane (80 mL × 3). The combined organic phases were washed with saturated sodium chloride solution (200 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was purified by silica gel column chromatography (dichloromethane / methanol = 1 / 0 to 15 / 1) to obtain compound 11-5. LCMS: 310.2 [M+1] + .

[0455] Step 4: Under a nitrogen atmosphere at -78°C, a solution of diisobutylaluminum hexane (1M, 19.4mL, 19.4mmol) was added dropwise to an anhydrous tetrahydrofuran (30mL) solution of compound 11-5 (3.0g, 9.70mmol). The reaction mixture was stirred at -78°C for 4 hours. The reaction was quenched with 60mL of water, extracted with dichloromethane (60mL × 3), and the combined organic phases were washed with 120mL of saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product. Purification was achieved by silica gel column chromatography (dichloromethane / methanol = 1 / 0 to 10 / 1) to give compound 11-6. LCMS: 268.2 [M+1] + . 1 H NMR (400MHz, CDCl3) δ = 6.76 (s, 1H), 4.72 (br d, J = 7.1Hz, 1H), 4.54 (s, 2H), 4.27-3.57 (m, 3H), 3.19 (br dd,J=5.2,16.6Hz,1H),2.75-2.75(m,1H),2.71(dd,J=7.9,16.5Hz,1H),2.31-2.19(m,1H),1.45(s,9H).

[0456] Step 5: Manganese dioxide (6.50 g, 74.8 mmol) was slowly added in portions to a mixed solution of compound 11-6 (2 g, 7.48 mmol) in dichloromethane (20 mL) and methanol (20 mL). The mixture was heated to 65 °C and stirred for 12 hours. After the reaction was complete, the reaction solution was cooled to room temperature, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was purified by silica gel column chromatography (dichloromethane / methanol = 1 / 0 to 15 / 1) to obtain compound 11-7. 1H NMR (400MHz, CDCl3) δ = 9.83 (s, 1H), 7.53 (s, 1H), 4.63 (br d,J=1.2Hz,1H),4.24-4.00(m,3H),3.29(dd,J=5.2,16.8Hz,1H),2.79(d d,J=8.4,16.8Hz,1H),2.39-2.25(m,1H),2.16-2.03(m,1H),1.46(s,9H).

[0457] Step 6: Under a nitrogen atmosphere, sodium acetate (417 mg, 5.09 mmol) and hydroxylamine hydrochloride (283 mg, 4.07 mmol) were added to a methanol (5 mL) solution of compound 11-7 (900 mg, 3.39 mmol). The reaction mixture was stirred at 25 °C for 1 hour. The solution was diluted with 20 mL of water and extracted with ethyl acetate (20 mL × 2). The combined organic phases were washed with 20 mL of saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain compound 11-8. LCMS: 281.2 [M+1] + .

[0458] Step 7: Under a nitrogen atmosphere at 0°C, slowly add Burgess reagent (1.15 g, 4.82 mmol) to a tetrahydrofuran (5 mL) solution of compound 11-8 (900 mg, 3.21 mmol). After the addition is complete, raise the temperature to 25°C and stir for 12 hours. Quench the reaction mixture with 5 mL of water, extract with ethyl acetate (20 mL × 2), wash the combined organic phases with 30 mL of saturated brine, dry to anhydrous sodium sulfate, filter, concentrate the filtrate under reduced pressure, and purify by silica gel column chromatography (petroleum ether / ethyl acetate = 20 / 1 to 1 / 1) to obtain compound 11-9. 1 H NMR (400MHz, CDCl3) δ = 7.37 (s, 1H), 4.79 (br d,J=6.8Hz,1H),4.19-3.97(m,3H),3.23(dd,J=5.2,16.8Hz,1H),2.75(d d,J=8.0,16.8Hz,1H),2.36-2.21(m,1H),2.18-2.06(m,1H),1.46(s,9H).

[0459] Step 8: Under a nitrogen atmosphere at 0°C, trimethyliodosilane (112 μL, 823 μmol) was slowly added to a dichloromethane (1 mL) solution of compound 11-9 (180 mg, 686 μmol), and the mixture was stirred at 0°C for 0.5 hours. The reaction solution was cooled to room temperature and concentrated to obtain crude hydroiodide of compound 11-10.

[0460] Step 9: Under a nitrogen atmosphere, N,N-diisopropylethylamine (401 mg, 3.10 mmol) was added to a solution of compound 11-10 (180 mg, crude hydroiodide) and compound 2-12 (148 mg, 745 μmol) in isopropanol (2 mL). The reaction mixture was heated to 80 °C and stirred for 12 hours. The reaction mixture was cooled to room temperature, diluted with 10 mL of water, and extracted with ethyl acetate (20 mL × 2). The combined organic phases were washed with 50 mL of saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 10 / 1 to 0 / 1) to obtain compound 11-11.

[0461] Step 10: Under a nitrogen atmosphere, palladium on carbon (100 mg, 5% purity) was added to a mixed solution of compound 11-11 (180 mg, 555 μmol) in dichloromethane (5 mL) and methanol (5 mL). Hydrogen was then purged three times, and the mixture was stirred for 0.5 hours under a hydrogen atmosphere (15 psi) at 25°C. The reaction solution was filtered, and the filtrate was concentrated under reduced pressure to obtain compound 11-12. LCMS: 295.2 [M+1] + .

[0462] Step 11: Under a nitrogen atmosphere at room temperature, trimethyl orthoacetate (131 mg, 1.09 mmol) and p-toluenesulfonic acid (9.36 mg, 54.4 μmol) were added sequentially to a mixed solution of compounds 11-12 (160 mg, 544 μmol) in toluene (2 mL) and tetrahydrofuran (2 mL). The reaction mixture was heated to 70 °C and stirred for 5 hours. The reaction mixture was cooled to room temperature, diluted with 10 mL of water, and extracted with ethyl acetate (20 mL × 2). The combined organic phases were washed with 50 mL of saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product. This crude product was purified by silica gel column chromatography (dichloromethane / methanol = 1 / 0-10 / 1) to obtain compound 11. LCMS: 319.1 [M+1] + .

[0463] Step 12: Compound 11 was separated by SFC (DAICL CHIRALPAK AD column (250 mm × 30 mm, 10 μm, mobile phase: A phase is supercritical CO2, B phase is ethanol / acetonitrile (4:1) (0.1% NH3H2O); gradient (B%): 60%) to obtain compound 11A and compound 11B.

[0464] Characterization of compound 11A: LCMS: 319.1 [M+1] + ; 1H NMR (400MHz, DMSO-d6) δ = 8.83 (s, 1H), 8.25 (d, J = 2.4Hz, 1H), 8.16 (s, 1H), 7.23 (d, J = 2.4Hz, 1H), 5.32-5.16 (m, 1H), 4. 42(brdd,J=4.8,12.8Hz,1H),4.26(dt,J=4.4,12.8Hz,1H),3.51-3.43(m,2H),2.93-2.81(m,1H),2.71(s,3H),2.39(br d,J=12.8Hz,1H). SFC detection (Chiralpak AD-3 column 50×4.6mm I.D., 3μm, mobile phase: A phase is supercritical CO2, B phase is ethanol (0.05% diethylamine); gradient (B%): 30%-60%), the retention time of compound 11A was 0.829 min, and the chiral purity was 100%.

[0465] Characterization of compound 11B: LCMS: 319.1 [M+1] + ; 1 H NMR (400MHz, DMSO-d6) δ = 8.83 (s, 1H), 8.25 (d, J = 2.4Hz, 1H), 8.16 (s, 1H), 7.23 (d, J = 2.4Hz, 1H), 5.33-5.16 (m, 1H), 4.42 (d d,J=4.8,12.8Hz,1H),4.26(dt,J=4.4,12.8Hz,1H),3.53-3.41(m,2H),2.86(dq,J=5.6,12.8Hz,1H),2.71(s,3H),2.39(br d,J=12.8Hz,1H). SFC detection (Chiralpak AD-3 column 50×4.6mm ID, 3μm, mobile phase: A phase is supercritical CO2, B phase is ethanol (0.05% diethylamine); gradient (B%): 30%-60%), the retention time of compound 11B was 1.884 min, and the chiral purity was 99.85%.

[0466] Example 12

[0467]

[0468] Under a nitrogen atmosphere, 1,8-diazabicyclo[5.4.0]undecane-7-ene (26.57 mg, 174.51 μmol) and iodomethane (18.58 mg, 130.88 μmol) were added to a solution of compound 10 (29 mg, 87.26 μmol) in N,N-dimethylformamide (1 mL). The reaction mixture was stirred at 60 °C for 1 hour. The reaction mixture was cooled to room temperature, filtered, and the filtrate was concentrated under reduced pressure to obtain a crude product. This crude product was then purified by reversed-phase column chromatography (column: Phenomenex Luna C18 150*25 mm*10 μm; mobile phase: H2O (0.225% formic acid)-acetonitrile; gradient (acetonitrile %): 0%-30%) to obtain compound 12. LCMS: 347.3. [M+H] + ; 1 H NMR (400MHz, DMSO-d6) δ = 9.65 (s, 1H), 8.75 (d, J = 2.0Hz, 1H), 8.51 (s, 1H), 7.86 (d, J = 2.0Hz, 1H), 6.15 (s, 1H), 5.48-5.29 (m, 1H), 4 .76(dd,J=6.0,12.1Hz,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).

[0469] Example 13: Preparation of the A-phase of compound 3

[0470] 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, then vacuum dried overnight at 40 °C to obtain crystal form A of compound 3. The XRPD, DSC, and TGA results are as follows: Figure 3 , 4 As shown in Figure 5.

[0471] Example 14: Preparation of the B crystal form of compound 3

[0472] Method 1: Compound 3 (10 mg) was added to solvent X (solvent X was 0.5 mL ethanol), stirred at 50 °C for 3 days, and then vacuum dried at 40 °C overnight to obtain the B crystal form of compound 3. The XRPD, DSC, and TGA results are as follows: Figure 6 , 7 As shown in Figure 8.

[0473] The solvent X may also be selected from ethanol / water (9 / 1, v / v), or solvent X may be selected from one or more mixed solvents selected from isopropanol, acetone, acetonitrile, ethyl acetate, tetrahydrofuran, methyl tert-butyl ether, 2-methyltetrahydrofuran, isopropyl acetate, dichloromethane, toluene, and 1,4-dioxane.

[0474] Method 2: Compound 3 (10 mg) was added to solvent Y (0.5 mL), stirred at room temperature for 3 days, and then vacuum dried overnight at 40 °C to obtain the B crystal form of compound 3. Solvent Y was selected from ethanol / water (9 / 1, v / v), or solvent Y was selected from one or more mixed solvents selected from ethanol, isopropanol, acetone, acetonitrile, ethyl acetate, tetrahydrofuran, 2-methyltetrahydrofuran, isopropyl acetate, dichloromethane, and 1,4-dioxane.

[0475] Method 3: Compound 3 (50 mg) was added to solvent Z (0.5–1 mL), and subjected to a temperature cycling process (5–55 °C, heating rate 1 °C; 55–5 °C, cooling rate 0.25 °C / min, 13 cycles), followed by vacuum drying at 40 °C overnight to obtain the B crystal form of compound 3. Solvent Z was selected from ethanol / water (9 / 1, v / v), or solvent Z was selected from one or more mixed solvents selected from ethanol, isopropanol, acetone, acetonitrile, tetrahydrofuran, and 1,4-dioxane.

[0476] Example 15: Preparation of the C-crystal form of compound 3

[0477] Compound 3 (100 mg) was added to methanol (0.5 mL), stirred at room temperature for 3 days, and then vacuum dried overnight at 40 °C to obtain the C crystal form of compound 3. The XRPD, DSC, and TGA results are as follows: Figure 9 , 10 As shown in Figure 11.

[0478] Example 16: Preparation of the D-crystal form of compound 3'

[0479] Compound 3 (100 mg) was added to water (2 mL), stirred at room temperature for 3 days, and then vacuum dried overnight at 40 °C to obtain the D crystal form of compound 3'. The XRPD, DSC, and TGA results are as follows: Figure 12 , 13 As shown in Figure 14.

[0480] Example 17: Solid Stability Study

[0481] Compound 3, crystal form B, was tested under the following conditions: 40℃ / 75%RH, 25℃ / 60%RH, high temperature 60℃, high humidity 90%RH, and illumination (total illuminance not less than 1.2 × 10⁻⁶). 6 Lux·hr, near-ultraviolet energy not less than 200 W·hr / m2 After being stored under a light source for one month, the XRPD spectrum showed no change in crystal form.

[0482] The D crystal form of compound 3' is stable under the following conditions: 40℃ / 75%RH, 25℃ / 60%RH, high temperature 60℃, and illumination (total illuminance not less than 1.2 × 10⁻⁶). 6 Lux·hr, near-ultraviolet energy not less than 200 W·hr / m 2 After being stored under a light source for one month, the XRPD spectrum showed no change in crystal form.

[0483] Both the B-type and D-type of compound 3' exhibit good solid stability under high temperature, high humidity, or light conditions.

[0484] Example 18: Hygroscopicity Study

[0485] The DVS test conditions are shown in Table 6. The DVS test results show that the B crystal form of compound 3 has a moisture absorption weight gain of 0.2314% at 80% RH. The DVS spectrum of the B crystal form of compound 3 is shown in [Table 6]. Figure 15 The D-crystal form of compound 3' showed a hygroscopic weight gain of 1.536% at 80% RH. The DVS spectrum of the D-crystal form of compound 3' is shown below. Figure 16 .

[0486] Hygroscopicity evaluation classification: Solubility: absorbs sufficient water to form a liquid; Extremely hygroscopic: ΔW% ≥ 15%; Hygroscopic: 15% > ΔW% ≥ 2%; Slightly hygroscopic: 2% > ΔW% ≥ 0.2%; No or almost no hygroscopicity: ΔW% < 0.2%.

[0487] Conclusion: The B crystal form of compound 3 and the D crystal form of compound 3' are slightly hygroscopic at 80% RH.

[0488] Reference compound 1

[0489]

[0490] The control compound 1 is Example 11 of patent WO2018067422A1.

[0491] Reference compound 2

[0492]

[0493] Following the preparation method of Examples 457 / 458 of WO2011086053A1, control compound 2 was prepared and then separated by SFC (column: DAICEL CHIRALPAK AD (250mm*30mm, 10μm, mobile phase: A phase is supercritical CO2, B phase is methanol (0.1% ammonia); gradient (B%): 30%) to obtain control compound 2A and control compound 2B.

[0494] Characterization of control compound 2A: LCMS: 257.3 [M+H] + ; 1 ¹H NMR (400MHz, DMSO-d6) δ=11.85(s,1H),8.47(s,1H),7.48(t,J=2.8Hz,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=11.6Hz,1H),1.89(s,2H); Detected by SFC (column: Chiralpak AD-350*4.6mm) ID, 3 μm, mobile phase: phase A is supercritical CO2, phase B is methanol (0.05% diethylamine); gradient (B%): 5%-40%), retention time of control compound 2A is 1.743 min, chiral purity is 100%.

[0495] Characterization of control compound 2B: LCMS: 257.3 [M+H] + ; 1 ¹H NMR (400MHz, DMSO-d6) δ=11.84(s,1H),8.47(s,1H),7.48(t,J=2.6Hz,1H),6.81(s,1H),4.80-4.43(m,1H),4.17-3.86(m,3H),3.75-3.57(m,1H),2.65(s,3H),2.59-2.51(m,1H),2.09(d,J=11.6Hz,1H),1.89(s,2H); Detected by SFC (column: Chiralpak AD-350*4.6mm) ID, 3 μm, mobile phase: phase A is supercritical CO2, phase B is methanol (0.05% diethylamine); gradient (B%): 5%-40%), the retention time of control compound 2B is 1.886 min, and the chiral purity is 97.86%.

[0496] Biological test data

[0497] Test Example 1: Kinase Activity Test

[0498] Enzyme activity assays were performed using Cisbio’s homogeneous time-resolved fluorescence resonance energy transfer (HTRF) technology to detect the inhibitory effects of the compounds on the activity of JAK1 / JAK2 / JAK3 / TYK2 JH1 kinases (JAKs).

[0499] 1. Experimental materials:

[0500] JAK1, JAK2, JAK3, and TYK2 kinase proteins were all purchased from Thermo Fisher Scientific. KinEASE TM The TK 20000 tests kit was purchased from Cisbio; the microplate reader was PerkinElmer, model Envision 2104 Multilabel Reader.

[0501] 2. Experimental methods:

[0502] (1) Preparation of reaction buffer: Take 10 ml as an example, prepare fresh on the day of the experiment. See Tables 7.1 to 7.3 for preparation parameters.

[0503] Table 7.1 JAK1 JH1 Reaction Buffer

[0504] Element Storage liquid concentration (mM) Final concentration (mM) Amount added (μL) 5× buffer solution 2000 <![CDATA[MgCl2]]> 1000 5 50 EGTA 455 0.625 13.7 SEB 2.5μM 0.06μM 240 Brij-35 (%) 5 0.01 20 DTT 1000 1 10 <![CDATA[H2O]]> 7666.3 total 10000

[0505] Table 7.2 JAK2 / 3JH1 Reaction Buffer

[0506] Element Storage liquid concentration (mM) Final concentration (mM) Amount added (μL) 5× buffer solution 2000 <![CDATA[MgCl2]]> 1000 5 50 DTT 1000 1 10 <![CDATA[H2O]]> 7940 total 10000

[0507] Table 7.3 TYK2 JH1 Reaction Buffer

[0508] Element Storage liquid concentration (mM) Final concentration (mM) Amount added (μL) 5× buffer solution 2000 <![CDATA[MgCl2]]> 1000 5 50 <![CDATA[MnCl2]]> 1000 1 10 SEB 2.5μM 0.0125μM 50 DTT 1000 1 10 <![CDATA[H2O]]> 7880 total 10000

[0509] (2) Experimental procedure

[0510] Step a, Pre-incubation of the compound with the kinase: Prepare 2×JAK1 / JAK2 / JAK3 / TYK2 JH1 enzyme solutions (final enzyme concentrations of 0.133 ng / μL, 0.0225 ng / μL, 0.224 ng / μL, and 0.25 ng / μL, respectively) using the corresponding reaction buffers. Transfer the compound at 100 nL per well to a 384-well plate using an automated micropipette system (DMSO is used instead of DMSO for negative and positive controls). Add 5 μL of 2×JAK1 / JAK2 / JAK3 / TYK2 JH1 enzyme solution (compound wells and positive control wells) or reaction buffer (negative control wells) to each well, centrifuge to mix, and incubate at 25°C for 15 minutes.

[0511] Step b, Enzyme Reaction: Prepare a mixed solution of 2×TK-Sub-biotin substrate and ATP using the appropriate reaction buffer (for JAK1, JAK2, JAK3, and TYK2 enzyme reactions, the final concentrations of TK-Sub-biotin substrate are 5 μM, 2 μM, 0.6 μM, and 1 μM, respectively; the final concentrations of ATP are 2 μM, 0.7 μM, 0.85 μM, and 0.1 μM, respectively). After incubation in step a, add 5 μL of the 2×TK-Sub-biotin substrate and ATP mixed solution to each well of the 384-well plate, centrifuge to mix, and react at 25°C for 45 minutes (JAK1 / JAK2 JH1) or 60 minutes (JAK3 / TYK2 JH1).

[0512] Step c, Detection: Prepare a detection mixture of TK Antibody-Eu and streptavidin-XL665 using detection buffer (final concentration of TK Antibody-Eu: 0.25×, final concentration of streptavidin-XL665: 1 / 16 of the final concentration of TK-Sub-biotin substrate). After incubation in step b, add 10 μL of the detection mixture to each well of the 384-well plate, centrifuge to mix, and react at 25°C for 60 minutes (JAK1 / JAK2 JH1) or 120 minutes (JAK3 / TYK2 JH1). After the reaction, read the fluorescence values ​​on an Envision 2104 Multilabel Reader (excitation at 340 nm, detection of emission at 665 nm and 615 nm; the 665 nm / 615 nm fluorescence ratio is the raw data of the reaction signal in the well).

[0513] 3. Data Processing and Analysis

[0514] First, the average response signals of the positive and negative control wells were calculated separately. Then, the inhibition rate of each compound well was calculated using the formula: "Inhibition rate per well = (1 - (Single well signal value - Average negative control signal value) / (Average positive control signal value - Average negative control signal value)) × 100%". Next, the concentration and corresponding inhibition rate data were imported into XLfit software. Using the Dose Response One Site205 model in the software, a four-parameter method was employed to fit the inhibition rate-concentration curve, and the IC50 of the compound was calculated. 50 value.

[0515] The experimental results are shown in Table 7.4. Where A represents IC. 50 ≤1nM(A+ further represents IC) 50 ≤0.2nM), B represents 1nM <IC 50 ≤20nM(B+ further indicates 1nM) <IC 50≤10nM), C represents 20nM <IC 50 ≤100nM (C+ further indicates 20nM) <IC 50 ≤50nM), D represents 100nM <IC 50 ≤500nM (D+ further indicates 100nM) <IC 50 ≤200nM), E represents IC 50 >500nM. The selectivity factor of JAK1-JAK2 is JAK2 IC. 50 Value and JAK1 IC 50 The ratio of values, the selectivity factor of JAK1-JAK3 is JAK3 IC 50 Value and JAK1 IC 50 The ratio of values.

[0516] Experimental results show that the compounds of this invention exhibit strong inhibitory activity against JAK1 kinase, significantly superior to the control compounds. Specifically, compounds 4 and 5 show significantly higher IC50 values. 50 All were ≤1 nM (5 times better than control compound 1, 17.5 times better than control compound 2A, and 55.3 times better than control compound 2B). The IC50 values ​​for compounds 1A, 3, and 6 were... 50 All were ≤0.2 nM (superior to control compound 1 by 10, 25, and 9.5 times respectively, superior to control compound 2A by more than 30 times, and superior to control compound 2B by more than 100 times).

[0517] The compounds of this invention exhibit strong inhibitory activity against TYK2 kinase, significantly superior to the control compounds. Specifically, compound 1A has an IC50 value of [missing value]. 50 ≤10 nM (15 times better inhibitory activity than control compound 1, 17.2 times better inhibitory activity than control compound 2A, and more than 30 times better inhibitory activity than control compound 2B), IC50 of compound 3 50 ≤1nM (15 times better than control compound 1, more than 100 times better than control compound 2A, and more than 100 times better than control compound 2B).

[0518] The compounds of this invention exhibit significantly weaker inhibitory activity against JAK2 and JAK3 than against JAK1, demonstrating higher selectivity, and their selectivity folds are significantly superior to those of the control compounds. Specifically, the JAK1-JAK2 selectivity folds of compounds 1A and 6 are 1.6 times and 2.9 times that of control compound 1, respectively; the JAK1-JAK2 selectivity folds of compounds 1A, 2A, 3, 4, 5, and 6 are 2.3 to 12.5 times that of control compound 2A and 3.3 to 19.3 times that of control compound 2B; the JAK1-JAK3 selectivity folds of compounds 1A, 2A, 3, 4, 5, and 6 are 8.8 to 50.3 times that of control compound 2A and 19.2 to 109.5 times that of control compound 2B.

[0519] Table 7.4 Results of kinase activity tests of the compounds of the present invention (IC50) 50 (nM)

[0520]

[0521] Conclusion: The compounds of this invention have strong inhibitory activity against JAK1 and TYK2 kinases, and their inhibitory activity against JAK2 and JAK3 is significantly weaker than that against JAK1, demonstrating high selectivity.

[0522] Test Example 2: PBMC Test

[0523] 1. Assay for TYK2 inhibitory activity in human peripheral blood mononuclear cells (PBMCs)

[0524] Human PBMCs were seeded into 96-well plates and incubated at 37°C for 1 hour. Then, different concentrations of the test compounds were added and incubated at 37°C for 1 hour. IFN-α (final concentration 30 ng / mL) and CD3 antibody (1.5 μL per well) were added and incubated at 37°C for 30 minutes. The mixture was then transferred to 96-well deep-plates, and 1 mL of pre-warmed 1×Lyse / Fix buffer (37°C) was added and incubated at 37°C for 10 minutes. The plates were centrifuged at 600g for 5 minutes, washed twice with PBS, and then incubated at 4°C for 30 minutes with Perm buffer III (400 μL per well). The plates were centrifuged at 600g for 5 minutes, and then washed twice with 1 mL of Staining buffer (DPBS + 0.2% BSA + 1 mM EDTA). Mouse anti-human Phospho-STAT5 (pY694) antibody was then incubated in Staining buffer. Dilute 200-fold in the buffer, add 100 μL to each well, mix well, and incubate at room temperature for 40 minutes. Add 1 mL of staining buffer per well, centrifuge at 600g for 5 minutes, and wash twice. Discard the supernatant, resuspend the cell pellet in 200 μL of staining buffer, and then load it for analysis in a Beckman CytoFlex flow cytometer.

[0525] 2. JAK1 inhibitory activity assay in human peripheral blood mononuclear cells (PBMCs)

[0526] Human PBMCs were seeded into 96-well plates and incubated at 37°C for 1 hour. Then, different concentrations of the test compounds were added and incubated at 37°C for 1 hour. IL-6 (final concentration 50 ng / mL) and CD3 antibody (1.5 μL per well) were added and incubated at 37°C for 30 minutes. The mixture was then transferred to 96-well deep-plates, and 1 mL of pre-warmed 1×Lyse / Fix buffer (37°C) was added and incubated at 37°C for 10 minutes. The plates were centrifuged at 600g for 5 minutes, washed twice with PBS, and then Perm buffer III (400 μL per well) was added and incubated at 4°C for 30 minutes. The plates were centrifuged at 600g for 5 minutes, and then 1 mL of sealing buffer (DPBS + 0.2% BSA + 1 mM EDTA) was added and washed twice. Alexa Fluor 647 anti-STAT3 Phospho (Tyr705) antibody antibody was then added to the sealing buffer. Dilute 100-fold in the buffer, add 100 μL to each well, mix well, and incubate at room temperature for 40 minutes. Add 1 mL of staining buffer per well, centrifuge at 600g for 5 minutes, and wash twice. Discard the supernatant, resuspend the cell pellet in 200 μL of staining buffer, and then load it for analysis in a Beckman CytoFlex flow cytometer.

[0527] 3. JAK2 inhibitory activity assay in human peripheral blood mononuclear cells (PBMCs)

[0528] Human PBMCs were seeded into 96-well plates and incubated at 37°C for 1 hour. Then, different concentrations of the test compounds were added and incubated at 37°C for 30 minutes. GM-CSF (final concentration 10 ng / mL) and CD14 antibody (1.0 μL per well) were added and incubated at 37°C for 20 minutes. The mixture was then transferred to 96-well deep-well plates, and 1 mL of pre-warmed 1×Lyse / Fix buffer (37°C) was added and incubated at 37°C for 10 minutes. The plates were centrifuged at 600g for 5 minutes, washed twice with PBS, and then incubated at 4°C for 30 minutes with Perm buffer III (400 μL per well). The plates were centrifuged at 600g for 5 minutes, and then washed twice with 1 mL of Staining buffer (DPBS + 0.2% BSA + 1 mM EDTA). Mouse anti-human Phospho-STAT5 antibody was diluted 200-fold in Staining buffer, and 100 μL was added to each well. The mixture was incubated at room temperature for 40 minutes. Finally, 1 mL of Staining buffer was added to each well. Centrifuge at 600g for 5 minutes and wash twice; discard the supernatant and resuspend the cell pellet in 200μL of Staining buffer, then load and analyze in a Beckman CytoFlex flow cytometer.

[0529] The experimental results are shown in Table 8. Where A represents IC. 50 ≤10nM, B represents 10nM <IC 50 ≤100nM, where C represents 100nM <IC 50 ≤500nM, D represents IC 50 >500 nM. Results showed that compounds 1A and 3 exhibited strong inhibitory activity against human peripheral blood mononuclear cells (PBMCs) TYK2, which was 5.6-fold and 12.3-fold superior to the inhibitory activity of control compound 1, respectively; compounds 1A and 3 also exhibited strong inhibitory activity against human peripheral blood mononuclear cells (PBMCs) JAK1, which was 4.5-fold and 11.9-fold superior to the inhibitory activity of control compound 1, respectively. The inhibitory activities of compounds 1A and 3 against human peripheral blood mononuclear cells (PBMCs) TYK2 and JAK1 were significantly superior to those against reference compound 1.

[0530] Table 8 PBMC Test Results

[0531]

[0532] Conclusion: The compounds of this invention have strong inhibitory activity against both JAK1 and TYK2, but weak inhibitory activity against JAK2, and exhibit high selectivity.

[0533] Test Example 3: Effects of LPS on the secretion of inflammatory factors in a microglial cell inflammation model

[0534] Mouse microglia BV-2 cell lines were resuscitated and cultured. After observing cell growth to 70-80% under a microscope, test compounds (concentrations of 0.3 μM, 1.5 μM, and 7.5 μM) were added. One hour later, lipopolysaccharide (LPS) solution was added to a final LPS concentration of 100 ng / mL to construct an LPS-induced cell inflammation model. After 24 hours, the cell supernatant was collected, and the inflammatory factors interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), and chemokine 2 (CCL-2) were detected using enzyme-linked immunosorbent assay (ELISA).

[0535] The experimental results are shown in Table 9. The results showed that, compared with the model group, the levels of inflammatory factors (IL-6, TNF-α, and TNF-α) were significantly reduced after adding different concentrations of compound 3, and the reduction was dose-dependent; among them, the inhibition rate of IL-6 by compound 3 at concentrations of 1.5 μM and 7.5 μM both exceeded 100%.

[0536] Table 9: Results of the effects of the compounds of the present invention on the secretion of inflammatory factors

[0537]

[0538] Conclusion: The compounds of this invention can significantly reduce the secretion of inflammatory factors (IL-6, TNF-α and CCL-2) in a dose-dependent manner.

[0539] Test Example 4: Plasma Protein Binding Assay (Balanced Dialysis Method)

[0540] Frozen plasma from CD-1 mice, Sprague-Dawley rats, beagles, cynomolgus monkeys, and humans was thawed in running, cold tap water. After complete thawing, the plasma was centrifuged at 3220×g for 5 minutes to remove suspended solids and precipitates. 597 μL of blank plasma from each species was taken, and 3 μL of the working solution of the test sample or control was added and mixed thoroughly to obtain plasma samples (n=1) with a concentration of 2 μM for both the test sample and control. The concentration of DMSO in the organic phase was 0.5%. The samples were thoroughly mixed before proceeding to the next step.

[0541] Transfer 50 μL of plasma samples from the test sample and control sample into a sample receiving plate (n=3), immediately add 50 μL of blank PBS, and then add 600 μL of stop solution to the T0 samples of the test sample and control sample respectively, and store at 2-8℃, waiting to be processed together with other dialysis samples.

[0542] Add 100 μL of test sample and control plasma sample to the administration end of each dialysis well (n=3), and add 100 μL of blank PBS to the receiving end of the corresponding dialysis well. Place the dialysis plate in a 5% CO2 incubator and incubate at 37°C with shaking at approximately 100 rpm for 4 hours.

[0543] After dialysis, 50 μL of dialysis-processed PBS and dialysis-processed plasma (n=3) were transferred to a new 96-well plate (sample receiving plate). The corresponding volume of blank plasma or PBS was added to each sample to make a final volume of 100 μL per well, with a plasma to PBS volume ratio of 1:1. All samples were analyzed by LC-MS / MS after protein precipitation.

[0544] The experimental results showed that the free proportion of all tested compounds was greater than 30% in the plasma of each animal and greater than 45% in human plasma, with no significant species differences. The higher proportion of free drug is beneficial to the efficacy of the drug in vivo.

[0545] Conclusion: The compounds of this invention exhibit high concentrations of free drug in the plasma of different species, demonstrating good drug-like properties.

[0546] Test Example 5: Liver Microsomal Stability

[0547] The test substance was prepared into a 10 mM DMSO solution with DMSO, and then diluted to 100 μM with 100% acetonitrile to obtain the working solution (organic phase content: 99% acetonitrile, 1% DMSO).

[0548] Prepare two 96-well incubation plates, and name them T60 incubation plate and NCF60 incubation plate respectively.

[0549] Add 445 μL of microsomal working solution (liver microsomal protein concentration of 0.56 mg / mL) to both T60 and NCF60 incubation plates, and then place the incubation plates in a 37°C water bath for pre-incubation for about 10 minutes.

[0550] After the pre-incubation is completed, add 5 μL of the working solution of the test sample or control compound to the T60 incubation plate and the NCF60 incubation plate respectively, and mix well.

[0551] To initiate the reaction, add 50 μL of potassium phosphate buffer to each well of the NCF60 incubation plate. In the T0 stop plate, add 180 μL of stop solution (containing 200 ng / mL tolbutamide and 200 ng / mL labenoyl in acetonitrile) and 6 μL of NADPH regeneration working solution. Transfer 54 μL of sample from the T60 incubation plate to the T0 stop plate (T0 sample generation). In the blank plate, add only 54 μL of microsomal working solution, 6 μL of NADPH regeneration working solution, and 180 μL of stop solution. To initiate the reaction, add 44 μL of NADPH regeneration working solution to each well of the T60 incubation plate. Therefore, in the sample of the test or control compound, the final reaction concentrations of the 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.

[0552] After incubation for appropriate times (e.g., 5, 15, 30, 45, and 60 minutes), add 180 μL of stop solution (containing 200 ng / mL tolbutamide and 200 ng / mL labenoyl in acetonitrile) to each well of the stop plate. Then, remove 60 μL of sample from the T60 or NCF60 incubation plate to terminate the reaction.

[0553] All sample plates were shaken well and centrifuged at 3220×g for 20 minutes. Then, 80 μL of the supernatant from each well was diluted to 240 μL of pure water for liquid chromatography-tandem mass spectrometry analysis. The in vitro elimination rate constant ke for the test and control compounds was calculated by converting the ratio of the compound's peak area to the internal standard peak area into a residual percentage using the formula below. The in vitro intrinsic clearance rate of liver microsomes (CL) was calculated using ke. int (mic)), the calculation formula is: CL int (mic) = 0.693 / T 1 / 2 / Microsomal protein content (microsomal concentration during incubation, mg / mL). Experimental results are shown in Table 10.

[0554] Table 10 Results of liver microsomal stability tests of the compounds of this invention

[0555]

[0556] Conclusion: The compounds of this invention exhibit excellent stability in various liver microsomes.

[0557] Test Example 6: Hepatocyte Stability

[0558] Prepare several 96-well sample precipitate plates, named T0, T15, T30, T60, T90, T0-MC, T90-MC, and blank matrix, respectively. Preheat the resuscitation and incubation media in a 37°C water bath. Remove the frozen hepatocytes from the liquid nitrogen container and immediately immerse them in the 37°C water bath (approximately 90 seconds). After the frozen portion has thawed and loosened, pour them into centrifuge tubes containing 40 mL of resuscitation media, gently inverting to resuspend the cells in the resuscitation media. Centrifuge at 100 × g for 5 minutes at room temperature, remove the supernatant, resuspend the hepatocytes in an appropriate volume of incubation media, and calculate cell viability using trypan blue staining. Add 198 μL of the hepatocyte suspension (0.51 × 10⁻⁶ m³ / h) to the centrifuge tubes. 6 (cells / mL) were added to the preheated incubation plate. For the culture medium control group, 198 μL of incubation medium without hepatocytes was added to the T0-MC and T120-MC incubation plates. All incubation plates were pre-incubated in a 37°C incubator for 10 minutes.

[0559] Then add 2 μL of the test sample and control compound working solution, mix well, and immediately place the incubation plate in the shaker inside the incubator and start the timer to begin the reaction. Prepare two replicate samples for each time point of each compound. The incubation conditions are 37°C, saturated humidity, and 5% CO2.

[0560] In the test system, the final concentration of the test sample was 1 μM, the final concentration of the control was 3 μM, and the final concentration of hepatocytes was 0.5 × 10⁻⁶. 6 The final concentration of total organic solvent was 0.96%, with DMSO at a final concentration of 0.1%. At the end of incubation at the corresponding time points, the incubation plate was removed, and 25 μL of the mixture of compound and control compound with cells was added to a sample plate containing 125 μL of stop solution (acetonitrile solution containing 200 ng / mL tolbutamide and labetalol). For blank sample plates, 25 μL of incubation medium without hepatocytes was added directly. After sealing all sample plates, they were shaken at 600 rpm for 10 minutes on a shaker, followed by centrifugation at 3220 × g for 20 minutes. The supernatants of the test and control samples were diluted with ultrapure water at a ratio of 1:3. All samples were analyzed by LC / MS / MS after mixing. The experimental results are shown in Table 11.

[0561] Table 11 Results of hepatocyte stability test of the compounds of the present invention

[0562]

[0563] Conclusion: The compounds of this invention exhibit good stability in various types of hepatocytes and show moderate or slow metabolism.

[0564] Test Example 7: In vitro MDCKII-MDR1 monolayer cell permeability test

[0565] This experiment used the MDR1-MDCK II cell line, authorized by the Piet Borst Laboratory of the Netherlands Cancer Institute, as an in vitro model for permeability assessment. It is a Madin-Darby canine kidney cell (MDCK II) transfected with the human multidrug resistance gene (MDR1). This cell line can predict the permeability of compounds in barriers with high efflux effects, such as the duodenum, blood-brain barrier, hepatocytes, and nephrons.

[0566] MDR1-MDCK II cells were seeded into Transwell-96-well cell plates. After the cells formed a complete monolayer membrane structure, transport experiments were performed. The drug was tested at a concentration of 2.00 μM. HBSS solution containing 10.0 mM HEPES (pH 7.40 ± 0.05) was used as the transport buffer. Bidirectional permeability was studied in the MDR1-MDCK II cell model, and incubation was performed at 37°C and 5% CO2 for 150 minutes. A fluorescent yellow assay was used to determine the integrity of the cell monolayer membrane, and low and high passive permeability controls, as well as a positive substrate for P-gp transport, were included in the experiment. After incubation, samples from the donor and receiver chambers were transferred and extracted with acetonitrile containing a suitable internal standard (IS). Proteins were precipitated by centrifugation at 3220 × g for 20 minutes, and the supernatant was diluted with ultrapure water (if necessary). Finally, the samples were analyzed using liquid chromatography-tandem mass spectrometry (LC-MS / MS) with appropriate MRM conversions for the analyte and internal standard. Calculate the apparent permeability coefficient (P) according to the following equation. app (cm / s) and efflux ratio (ER). The calculation formula is as follows:

[0567]

[0568] Among them, V R The volume of the receiving solution is 0.075 mL on side A and 0.25 mL on side B; Area is the relative surface area of ​​the cell monolayer (0.0804 cm²). 2 Time is the incubation time (9000s); C0 is the peak area ratio of the drug-attributed compound; C R These represent the peak area ratios of the compounds at the dosing and receiving ends, respectively. Experimental results show that the solvent recovery rates of the compounds tested in this invention are all greater than 90%, specifically P... app The ER data are shown in Table 12.

[0569] Table 12 Results of MDCKII-MDR1 cell permeability test of the compounds of the present invention.

[0570]

[0571] Conclusion: The compounds of this invention showed high recovery rates in the permeability experiment of MDCKII-MDR1 monolayer cells, exhibiting high permeability and low efflux.

[0572] Test Example 8: Study on Inhibition of Cytochrome P450 Enzyme (CYP)

[0573] 1. Purpose of the test: To determine the inhibitory effect of the test compound on the activity of human liver microsomal cytochrome P450 isoenzymes (CYP1A2, CYP2C9, CYP2C19, CYP2D6 and CYP3A4).

[0574] 2. Experimental Methods

[0575] The test compound (10.0 mM) was serially diluted to prepare working solutions (100 × final concentration), with working solution concentrations of 5.00, 1.50, 0.500, 0.150, 0.0500, 0.0150 and 0.00500 mM. Simultaneously, working solutions were prepared for mixtures of positive inhibitors and specific substrates of P450 isoenzymes (CYP1A2, CYP2C9, CYP2C19, CYP2D6 and CYP3A (using midazolam as a probe substrate)). Human liver microsomes stored at below -60°C were thawed on ice until completely dissolved, then diluted with potassium phosphate buffer (PB) to prepare a working solution of a specific concentration (0.253 mg / mL).

[0576] Add 20.0 μL of substrate mixture to the reaction plate (add 20.0 μL of PB to the blank well), then add 158 μL of human liver microsome working solution to the reaction plate. Place the reaction plate on ice and set aside. At this time, add 2.00 μL of each concentration of the test compound (N=1) and specific inhibitor (N=2) to the corresponding well. For the inhibitor-free group (no test compound or positive inhibitor), add the corresponding organic solvent as the control sample (the test compound control sample is DMSO:MeOH = 1:1, and the positive control samples are all DMSO:MeOH = 1:9). Preheat the reaction plate in a 37°C water bath. After incubation for 10 min, 20.0 μL of coenzyme factor (NADPH) solution was added to the reaction plate and incubated in a 37°C water bath 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 the supernatant was added to 100 μL of water to dilute the sample. Finally, the plate was sealed, shaken for 10 min to mix, and then detected by LC-MS / MS.

[0577] Experimental results showed that compounds 1A, 2A, and 3 had an IC50 inhibitory effect on human liver microsomal cytochrome P450 isoenzymes (CYP1A2, CYP2C9, CYP2C19, CYP2D6, and CYP3A4). 50 All are greater than 30 μM.

[0578] Conclusion: The compounds of this invention do not have a significant inhibitory effect on the five major cytochrome P450 enzymes in human liver microsomes, and the risk of drug-drug interactions is low.

[0579] Test Case 9: PXR / CAR / AhR Induction Study

[0580] PXR assays used HepG2-human PXR+CYP3A4 stable cell line; CAR assays used HepG2 cell line; AhR assays used HepG2-Lucia cell line. TM AhR cells are stable cell lines.

[0581] First, the test compound (30.0 mM) was serially diluted to prepare working solutions (100 × final concentration), with working solution concentrations of 3.00, 1.00, 0.333, 0.111, 0.0370, 0.0123, 0.00412, 0.00137 and 0.000457 mM.

[0582] CAR induction experimental method:

[0583] CAR assays were performed using the HepG2 cell line. After cell resuscitation, cells were passaged and cultured until the cell density reached 90%. Cells were then digested and counted at 3*102. 5 Cells / mL, 20 wells per compound (n=2), 100 μL / well, seeded in 96-well plates. Incubate at 37°C for 24 hours. Transfect CYP2B6 and CAR into cells, shake well, and incubate overnight. Discard the culture medium in the cell culture plate, and add 90 μL of culture medium again. Take 3 μL of each working solution and dilute 30 times with 87 μL of EMEM medium, mix well, and use as the drug to be added; take 10 μL of the drug to be added to the cell culture plate to make the high dose = 100 μM. DMSO is used as a negative control, with a DMSO content of 0.3%. Incubate at 37°C for 24 hours. Remove the cell culture plate from the incubator and place it at room temperature for 30 min to allow the culture plate temperature to equilibrate to room temperature. Equilibrate the Bright-Lite Luciferase Assay System reagent to room temperature, add 50 μL to each well, shake at 200 rpm for 3 min, and read the Luciferase signal value on a microplate reader (BMG). Drawing EC using GraphPadPrism 8 50Line graph.

[0584] PXR induction experimental method:

[0585] The PXR experiment used a HepG2-human PXR+CYP3A4 stable transgenic cell line; when the cell density reached 90%, the cells were digested and counted at 3*102. 5 Cells / mL, 20 wells per compound (n=2), 100 μL / well, seeded in 96-well plates; incubated at 37°C for 24 hours. Discard the culture medium from the cell culture plate, then add 90 μL of culture medium again. Take 3 μL of each working solution and dilute 30-fold with 87 μL of EMEM medium, mix well, and use as the drug to be added; add 10 μL of the drug to be added to the cell culture plate to make Highdose = 100 μM. DMSO is used as a negative control, with a DMSO concentration of 0.3%. Incubate at 37°C for 24 hours. Remove the cell culture plate from the incubator and let it equilibrate to room temperature for 30 min. Equilibrate the Bright-Lite Luciferase Assay System reagent to room temperature, add 50 μL to each well, shake at 200 rpm for 3 min, and read the Luciferase signal value on a microplate reader (BMG). Plot EC using GraphPad Prism 8. 50 Line graph.

[0586] AhR induction experimental method:

[0587] AhR testing uses HepG2-Lucia TM AhR cells were stably converted to a cell line and passaged. When the cell density reached 90%, cells were digested and counted. 4000 cells / well / 40 μL of each compound was used in 20 wells (n=2) for seeding in 384-well plates. The plates were incubated at 37°C for 24 hours. The culture medium in the cell culture plates was discarded, and 39 μL of medium was added again. Compound preparation: 4 μL of each working solution was diluted 7.5 times in 26 μL of LEMEM medium and mixed well. This was used as the drug to be added. 1 μL of the drug to be added was added to the cell culture plate to achieve a high dose of 100 μM. DMSO was used as a negative control, with a DMSO concentration of 0.3%. The plates were incubated at 37°C for 24 hours. The cell culture plates were removed from the incubator and allowed to equilibrate to room temperature for 30 minutes. QUANTI-Luc was then added. TM After equilibrating the Gold reagent to room temperature, transfer 4 μL of cell culture supernatant to a 384-well plate and add 20 μL of QUANTI-Luc. TMGold was centrifuged at 1000 rpm for 1 min, and the Luciferase signal value was read on a microplate reader (BMG). 5) EC was plotted using GraphPad Prism 8. 50 Line graph.

[0588] Experimental results showed that, at the highest experimental concentration (30 μM), the EC values ​​of compounds 1A and 3 were [missing information]. 50 All are greater than 30 μM.

[0589] Conclusion: The compounds of this invention did not show significant induction effects on CAR, PXR, and AhR, suggesting that the compounds of this invention have a very low risk of inducing major cytochrome P450 enzymes.

[0590] Test Example 10: hERG Test

[0591] CHO cells stably expressing hERG were cultured in 35 mm diameter cell culture dishes at 37°C in a 5% CO2 incubator, and passaged every 48 hours at a 1:5 ratio. On the day of the experiment, the cell culture medium was aspirated, the cells were rinsed once with extracellular fluid, and then 0.25% Trypsin-EDTA (Invitrogen) solution was added for digestion at room temperature for 3-5 minutes. The digestion solution was aspirated, the cells were resuspended in extracellular fluid, and then transferred to experimental dishes for electrophysiological recording.

[0592] The compound to be tested was prepared into a 20 mM stock solution using DMSO, and then serially diluted 3 times with DMSO, i.e., 10 μL was added to 20 μL of DMSO. Then, 10 μL of the serially diluted compound DMSO solution was added to 4990 μL of extracellular fluid. The final concentration to be tested was obtained by 500-fold dilution.

[0593] CHO cells stably expressing the hERG potassium channel were used to record hERG potassium channel currents at room temperature using whole-cell voltage-clamp technique. After obtaining whole-cell recordings, the cells were clamped at -100 mV. The step voltage to induce hERG potassium current (IhERG) was applied from -100 mV with a 2-s depolarization voltage to +20 mV, then repolarized to -50 mV for 1 second before returning to -100 mV. This voltage stimulation was applied every 5 seconds. Once the hERG potassium current stabilized (1 minute), the drug administration process began. Each test concentration of the compound was administered for at least 1 minute to steady state or for a maximum of 3 minutes, and at least 2 cells (n≥2) were tested for each concentration.

[0594] Data analysis was performed using pClamp and Excel software. The inhibition degree of different compound concentrations on the hERG potassium current (the peak hERG tail current induced at -50mV) was calculated using the following formula: Inhibition% = [1 – (I / I0)] × 100%. Wherein, Inhibition% represents the inhibition rate of the compound on the hERG potassium current, and I and I0 represent the amplitude of the hERG potassium current before and after drug administration, respectively. GraphPadPrism 8 was used for plotting and calculating IC. 50 .

[0595] Experimental results show that compound 3 has hERGIC 50 >40μM.

[0596] Conclusion: The compounds of this invention do not significantly inhibit hERG.

[0597] Test Example 11: PK Study

[0598] 1. Six male C57 / 6J mice were divided into two groups of three. The intravenous (iv) group received 2 mg / kg of medication in 5% DMSO / 10% Solutol / 85% Saline solution; the oral (po) group received 15 mg / kg of medication in 0.5% HPMC / water solution.

[0599] 2. Six male SD rats were divided into two groups of three. The intravenous (iv) group received 2 mg / kg of medication in 5% DMSO / 10% Solutol / 85% Saline solution; the oral (po) group received 10 mg / kg of medication in 0.5% HPMC / water solution (the po group of compound 1A received 0.5% HPMC / 0.2% Tween 80 / water solution).

[0600] 3. Six male beagles were divided into two groups of three animals each. The intravenous (iv) group received 0.5 mg / kg of medication in a solvent of 5% DMSO / 20% PEG400 / water; the oral (po) group received 2 mg / kg of medication in a solvent of 0.5% HPMC / 0.2% Tween 80 / water.

[0601] Whole blood was collected at 5 min (IV group only), 15 min, 30 min, 1 h, 2 h, 4 h, 8 h, and 24 h after drug administration. The whole blood was placed in anticoagulant tubes containing EDTA-K2 and centrifuged to prepare plasma. The concentration of the test substance molecules in the plasma was quantitatively detected by LC-MS / MS, and the pharmacokinetic parameters were calculated using PhoenixWinNonlin.

[0602] The experimental results are shown in Table 13. Note: CL represents the clearance rate, Vdss represents the distribution volume, and T represents the volume of distribution.1 / 2 For half-life, AUC 0-last C represents the area under the whole blood concentration-time curve from 0 to the last quantifiable time point. max To reach peak concentration; F represents bioavailability.

[0603] Table 13 PK test results of the compounds of the present invention

[0604]

[0605] Conclusion: The compounds of this invention exhibited high oral exposure and high oral bioavailability in mice, rats, and dogs, with little species variation and excellent pharmacokinetic properties.

[0606] Test Example 12: Brain-Tracking Test

[0607] 1. Twelve male C57 / 6J mice were divided into four groups of three. All animals were orally administered a 15 mg / kg dose of 0.5% HPMC aqueous solution. At 0.5, 2, 4, and 8 hours post-administration, one group of animals was sacrificed, and whole blood and brain tissue were collected. Whole blood was placed in anticoagulant tubes containing EDTA-K2 and centrifuged to prepare plasma. Brain tissue was homogenized with PBS. The concentrations of the test substance molecules in plasma and brain homogenate were quantitatively determined by LC-MS / MS (ng / mL in plasma, ng / g in brain homogenate), and PK parameters were calculated using Phoenix WinNonlin.

[0608] 2. Nine male SD rats were divided into three groups of three animals each. All animals were orally administered 10 mg / kg of a solution of 0.5% HPMC / 0.2% Tween 80 / water. At 0.5, 2, and 8 hours post-administration, one group of animals was sacrificed, and whole blood and brain tissue were collected. Whole blood was placed in anticoagulant tubes containing EDTA-K2 and centrifuged to prepare plasma. Brain tissue was homogenized with PBS. The concentrations of the test substance molecules in plasma and brain homogenate were quantitatively detected using LC-MS / MS, and the p-values ​​(PK) parameters were calculated using Phoenix WinNonlin. The experimental results are shown in Table 14.

[0609] Table 14 Results of brain penetration tests on the compounds of this invention

[0610]

[0611] Conclusion: The compounds of this invention exhibit good brain tissue distribution in both mice and rats.

[0612] Test Example 13: In vivo drug efficacy (Part 1)

[0613] Multiple sclerosis (MS) is a chronic inflammatory neurodegenerative disease of the central nervous system. Experimental autoimmune encephalomyelitis (EAE) induced by oligodendrocyte glycoprotein (MOG) is the most common animal model of MS in humans. This study aims to test the in vivo efficacy of the compounds of this invention in an EAE model.

[0614] Experimental methods:

[0615] 1. Model making:

[0616] C57BL / 6J mice (female, approximately 7 weeks old) were subcutaneously injected with 200 μL of MOG emulsion (Day 1) after an acclimatization period. The final MOG concentration in the emulsion was 1.5 mg / mL, and the concentration of Mycobacterium tuberculosis / complete Freund's adjuvant (CFA) solution was 4 mg / mL. Pertussis toxin (PTX, 1.0 μg / mL) was injected intraperitoneally on the day of MOG injection and the following day, at a dose of 250 μL per mouse. Body weight, disease incidence, and clinical scores were observed daily during the experiment. The scoring criteria were: 0, normal, no obvious symptoms; 1, tail weakness or mild hind limb weakness; 2, tail weakness and hind limb weakness, recoverable by passive turning; 3, unilateral hind limb hemiplegia, not recoverable by passive turning; 4, complete hind limb paralysis, forelimb paralysis or weakness, accompanied by urinary and fecal incontinence; 5, near-death state or death.

[0617] 2. Administration:

[0618] Oral administration via gavage began on Day 13 and continued until Day 24, twice daily (BID). Dosage: 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.

[0619] 3. Experimental Results:

[0620] Clinical scoring results of animals in the EAE model, as follows: Figure 1.1 As shown, the experimental results of animal weight change are as follows: Figure 1.2 As shown in the figure. Specifically, both compound 1A and compound 3 significantly reduced clinical scores in a dose-dependent manner. Animals in the compound 1A (30 mg / kg) and compound 3 (20 mg / kg) experimental groups showed no obvious symptoms from Day 13 to Day 24, and their clinical scores remained at 0. Regarding body weight, on day 22 of administration, the animals in the compound 1A and compound 3 groups recovered their body weight faster than the animals in the model group. The administration of these compounds significantly improved body weight, and the effect was dose-dependent.

[0621] Conclusion: The compounds of this invention exhibit significant therapeutic effects in the EAE pharmacodynamic model, and show a positive dose-effect correlation.

[0622] Test Example 14: In vivo drug efficacy (Part 2)

[0623] Experimental objective: To test the in vivo efficacy of the compound of this invention in an EAE model.

[0624] Experimental methods:

[0625] 1. Model making:

[0626] C57BL / 6J mice (female, approximately 7 weeks old) were subcutaneously injected with 200 μL of MOG emulsion (Day 1) after an acclimatization period. The final MOG concentration in the emulsion was 1.5 mg / mL, and the concentration of Mycobacterium tuberculosis / complete Freund's adjuvant (CFA) solution was 4 mg / mL. Pertussis toxin (PTX, 1.0 μg / mL) was injected intraperitoneally on the day of MOG injection and on the third day, at a dose of 250 μL per mouse. Body weight, disease incidence, and clinical scores were observed daily during the experiment. The scoring criteria were the same as in test case 13.

[0627] 2. Administration:

[0628] Oral administration via gavage began on Day 16 and continued until Day 30, twice daily (BID). Dosage: 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.

[0629] 3. Experimental Results:

[0630] Clinical scoring results of animals in the EAE model, as follows: Figure 2 As shown in Table 15, the area under the curve (AUC) results for Days 17-30 are presented. Specifically, both compound 1 and compound 3 significantly reduced clinical scores, with compound 3 showing a dose-dependent effect. The clinical scores and AUC of the experimental groups treated with compound 3 (7.5 mg / kg and 15 mg / kg) were lower than those of the group treated with compound 1 (30 mg / kg), indicating that compound 3 achieves comparable efficacy to compound 1 at a lower dose.

[0631] Table 15 Results of Area Under Curve

[0632]

[0633] Conclusion: The compounds of this invention exhibit excellent therapeutic effects in the EAE pharmacodynamic model, with a low effective dose and a positive dose-effect correlation.

Claims

1. A compound of formula (II), its stereoisomers or pharmaceutically acceptable salts thereof, in, It consists of double or single bonds, and the five-membered ring containing T1, T2, and T3 is an aromatic ring; 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 Where 1 represents the linking site with imidazole N, and 2 represents the linking 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 joined with ring B, and ring C is selected from one or more R's. c Substituted 5-10 nitrogen-containing heteroaryl groups; T is N, wherein N 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 respectively; R1 is -L1-R 11 ; L1 is selected from key, -CONR 12 -CO-, O, S, NR 12 and can be 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 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; R 11 Selected from H, D, F, Cl, Br, I, CN, and can be 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 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; 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; 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; 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; 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 Cycloalkyl, 3-8 membered heterocycloalkyl, phenyl and 5-6 membered heteroaryl, wherein C 1-4 Alkyl, C 2-4 alkenyl, C 2-4 alkynyl group, C 1-4 Alkoxy, C 1-4 Alkylthio, C 3-8 Cycloalkyl, 3-8 membered heterocycloalkyl, phenyl, and 5-6 membered heteroaryl groups are substituted with CN, and further optionally with one or more R groups. 4a replace; 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 The following groups are independently selected from H, D, F, Cl, Br, I, OH, NH2, CN, =O, and optionally substituted with one or more R groups: C 1-4 Alkyl, C 2-4 alkenyl, C 2-4 alkynyl group, C 1-4 Alkoxy, C 1-4 Alkylthio, C 3-8 Cycloalkyl, 3-8 membered heterocycloalkyl, phenyl and 5-6 membered heteroaryl; Or 2 Rs a1 Connected together, or two Rs b Connected together, or two Rs c Connected together, or R b and R c Linked together, they independently form the following groups optionally substituted with one or more R: C 3-8 Cycloalkyl, 4-8 membered heterocycloalkyl, phenyl or 5-6 membered heteroaryl; Each R is independently selected from H, D, F, Cl, Br, I, OH, NH2, CN, and the following groups optionally substituted by one or more F: 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.

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

3. The compound according to claim 1, its stereoisomers, or pharmaceutically acceptable salts thereof, wherein, Each R a1 The groups are independently selected from H, D, F, Cl, Br, I, OH, NH2, CN, and CH3, CH2CH3, vinyl, ethynyl, OCH3, OCH2CH3, cyclopropyl, and cyclobutyl, which are optionally substituted with one or more R groups; furthermore, each R group... a1 They are independently selected from H, D, F, Cl, CH3 and CF3, respectively.

4. The compound according to claim 1, its stereoisomers, or a pharmaceutically acceptable salt thereof, wherein, Each R b Each R c The following groups are independently selected from H, D, F, Cl, Br, I, OH, NH2, CN, =O, and optionally substituted with one or more R groups: CH3, CH2CH3, CH2CH2CH3, CH(CH3)2, vinyl, propenyl, ethynyl, propynyl, OCH3, OCH2CH3, OCH(CH3)2, SCH3, SCH2CH3, cyclopropyl, and cyclobutyl; furthermore, each R b Each R c They are independently selected from H, F, Cl, CH3 and CF3, respectively.

5. The compound according to claim 1, its stereoisomers, or pharmaceutically acceptable salts thereof, wherein, Each R 1a R 11a R 11b R 11c R 11d Each R 2a Each R 3a Each R 3b Each R 4a The following groups are independently selected from H, D, F, Cl, Br, I, OH, NH2, CN, =O, and optionally substituted with one or more R groups: CH3, CH2CH3, CH2CH2CH3, CH(CH3)2, vinyl, propenyl, ethynyl, propynyl, OCH3, OCH2CH3, OCH(CH3)2, SCH3, SCH2CH3, cyclopropyl, and cyclobutyl; furthermore, each R 1a R 11a R 11b R 11c R 11d Each R 2a Each R 3a Each R 3b Each R 4a The groups are independently selected from H, D, F, Cl, CN, =O, CH3, CFH2, CF2H, CF3, OCH3, OCF3 and cyclopropyl, respectively.

6. The compound according to claim 1, its stereoisomers, or a pharmaceutically acceptable salt thereof, wherein, L1 is selected from the bond, C(O), O, S, and optionally one or more R. 1a The following groups are substituted: C(O)NH, NH, CH2, CH2CH2, vinyl, ethynyl, propynyl, OCH2, OCH2CH2 and cyclopropyl, R 11 Selected from H, D, F, Cl, Br, I, CN, and can be selected by one or more R 1a The following groups are substituted: CH3, CH2CH3, CH2CH2CH3, CH(CH3)2, vinyl, ethynyl, propynyl, OCH3, OCH2CH3, OCH(CH3)2, SCH3, SCH2CH3, NHCH3, NHCH2CH3, N(CH3)2, cyclopropyl, cyclobutyl, oxacyclobutyl, and oxacyclopentyl; further, L1 is selected from the following groups: bond, O, S, and optionally substituted with one or more F or D: CH2, CH2CH2, OCH2, OCH2CH2, and cyclopropyl, R 11 Selected from H, D, F, Cl, Br, I, CN, and the following groups optionally substituted by one or more F or D: CH3, CH2CH3, CH2CH2CH3, CH(CH3)2, OCH3, OCH2CH3, OCH(CH3)2, cyclopropyl, cyclobutyl, oxacyclobutyl, oxacyclopentyl, 7. The compound according to claim 1, its stereoisomers, or a pharmaceutically acceptable salt thereof, 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 according to claim 1, its stereoisomers, or pharmaceutically acceptable salts thereof, wherein, R2 is H, and R3 is H.

9. The compound according to claim 1, its stereoisomers, or a pharmaceutically acceptable salt thereof, wherein, R4 is selected from CN, CH3, CH2CH3, CH2CH2CH3, CH(CH3)2, vinyl, propynyl, ethynyl, propynyl, OCH3, OCH2CH3, OCH(CH3)2, SCH3, SCH2CH3, cyclopropyl, and cyclobutyl, wherein the CH3, CH2CH3, CH2CH2CH3, CH(CH3)2, vinyl, propynyl, ethynyl, propynyl, OCH3, OCH2CH3, OCH(CH3)2, SCH3, SCH2CH3, cyclopropyl, and cyclobutyl are substituted with CN, and are further optionally substituted with one or more R. 4a Replace; furthermore, R4 is selected from CH2CN.

10. The compound according to claim 1, its stereoisomers, or pharmaceutically acceptable salts thereof, 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 according to claim 1, its stereoisomers, or a pharmaceutically acceptable salt thereof, wherein, Ring A2 is selected from 12. The compound according to claim 1, its stereoisomers, or pharmaceutically acceptable salts thereof, wherein, Structural unit Selected from Furthermore, structural units Selected from 13. The compound according to any one of claims 1 to 12, its stereoisomers, or its pharmaceutically acceptable salts, wherein the compound is selected from: in, m and n are independently selected from 0, 1, 2 and 3 respectively; R1 is -L1-R 11 ; L1 is selected from the bond, -CONH, -CO-, O, S, NH, and optionally 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 1-4 Alkoxy, C 1-4 Alkylthio, C 1-4 Alkylamino, C 3-8 Cycloalkyl and 3-8 membered heterocyclic alkyl groups; R 11 Selected from H, F, Cl, Br, I, CN and optionally one or more R 1a The following groups are substituted: C 1-4 Alkyl, C 2-4 alkenyl, C 2-4 alkynyl group, C 1-4 Alkoxy, -C 3-8 Cycloalkyl and 3-8 membered heterocyclic alkyl groups; R2 is selected from H, 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 3-8 Cycloalkyl and 3-8 membered heterocyclic alkyl groups; R3 is selected from H, 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 3-8 Cycloalkyl and 3-8 membered heterocyclic alkyl groups; R4 is selected from CN, CH3, CH2CH3, CH2CH2CH3, CH(CH3)2, vinyl, propynyl, ethynyl, propynyl, OCH3, OCH2CH3, OCH(CH3)2, SCH3, SCH2CH3, cyclopropyl, and cyclobutyl, wherein the CH3, CH2CH3, CH2CH2CH3, CH(CH3)2, vinyl, propynyl, ethynyl, propynyl, OCH3, OCH2CH3, OCH(CH3)2, SCH3, SCH2CH3, cyclopropyl, and cyclobutyl are substituted with CN, and are further optionally substituted with one or more R. 4a replace; Ring B is selected from one or more R. b Substituted monocyclic 5-7 member nitrogen-containing heterocyclic alkyl groups; Ring C is joined with ring B, and ring C is selected from one or more R's. c Substituted 5-6 nitrogen-containing heteroaryl groups; R a1 R 2a R 3a R 4a R 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.

14. The compound of formula (I-1) according to claim 13, its stereoisomers, or pharmaceutically acceptable salts thereof, wherein, m is selected from 0, 1, 2, and 3; n is selected from 1, 2, and 3; 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; R2 is selected from H, F, Cl, Br, I, and CH3; Each R3 is independently selected from H, F, Cl, Br, I, and CH3; R4 is 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 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.

15. The compound according to claim 13 or 14, its stereoisomer, or a pharmaceutically acceptable salt thereof is selected from: in, m and s are independently selected from 0, 1, 2 and 3 respectively; R1, R2, R3, R a1 R b R c As defined in claim 13 or 14.

16. The compound according to claim 15, its stereoisomers, or pharmaceutically acceptable salts thereof are selected from: in, 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; Each R 1a They were each independently selected from H, D, F, Cl, and CN; R4 is selected from CN, CH2CN, CH2CH2CN, CH(CH3)CN, C(CH3)2CN, 17. The compound of claim 16, its stereoisomers or pharmaceutically acceptable salts thereof, wherein R1 is selected from F, Cl, Br, I, CN and CH3 or OCH3 optionally substituted with 1, 2 or 3 F or D atoms.

18. The compound according to claim 17, its stereoisomers or pharmaceutically acceptable salts thereof, wherein R4 is selected from CH2CN.

19. The compounds shown in Table A and / or Table A1, their stereoisomers, or pharmaceutically acceptable salts thereof.

20. The B-type crystal of compound 3, Its Cu Kα radiation X-ray powder diffraction pattern has characteristic diffraction peaks at the following 2θ angles: 8.45±0.20°, 9.98±0.20° and 13.60±0.20°.

21. The B-type crystal according to claim 20, wherein the X-ray powder diffraction pattern of its Cu Kα radiation, represented by the 2θ angle, contains at least 5, 6, 7 or 8 diffraction peaks selected from the following: 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-type crystal according to claim 21, wherein the X-ray powder diffraction pattern of Cu Kα radiation has characteristic 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-type crystal according to any one of claims 20 to 22, further comprising any one of the following features: (1) Its XRPD map is basically as shown in Figure 6; (2) Its differential scanning calorimetry curve has an initial value of endothermic peak at 249.5±3℃; (3) Its DSC spectrum is basically as shown in Figure 7; (4) Its thermogravimetric analysis curve shows a weight loss of 0.25% in the range of 30.0±3℃ to 150.0±3℃; (5) Its TGA spectrum is basically as shown in Figure 8.

24. The use of the compound according to any one of claims 1 to 19, its stereoisomer or a pharmaceutically acceptable salt thereof, or the B crystal form according to any one of claims 20 to 23 in the preparation of a medicament for treating diseases related to JAK1 / TYK2 inhibitors.

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