Pyridone derivatives and medical use thereof

Abstract

The present invention provides pyridone derivatives and the medical use thereof, and particularly relates to compounds shown as general formula (I) or stereoisomers, racemates, deuterated substances, solvates, prodrugs, metabolites, pharmaceutically acceptable salts or eutectic crystals thereof, and an intermediate thereof and a preparation method therefor, and the use thereof in the preparation of a drug for treating diseases related to the activity or expression level of integrin α4β7.

Description

A pyridone derivative and its application in medicine

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS

[0002] The present invention claims all priority rights of Chinese patent application 202410188785.3 filed on February 20, 2024, Chinese patent application 202410292419.2 filed on March 14, 2024, Chinese patent application 202410456185.0 filed on April 16, 2024, Chinese patent application 202410707588.8 filed on June 3, 2024, and Chinese patent application 202411137105.1 filed on August 19, 2024, and the entire contents of the above applications are incorporated herein by reference. Technical Field

[0003] The present invention relates to a compound described by general formula (I) or its stereoisomers, racemates, deuterated substances, solvates, prodrugs, metabolites, pharmaceutically acceptable salts or cocrystals, as well as intermediates and preparation methods, and use of the compounds in preparing drugs for treating diseases related to integrin α4β7 activity or expression. Background Art

[0004] The integrin family consists of dimers of two subunits: α (120-185 kDa) and β (90-110 kDa). There are 18 α subunits and 8 β subunits in mammals. Different combinations of these subunits can form over 20 different integrins. α4β7 is a member of the integrin family. Intestinal inflammatory diseases associated with α4β7 include Crohn's disease and ulcerative colitis. The primary ligand for α4β7 is mucosal addressin cell adhesion molecule-1 (MADCAM-1). MAdCAM-1 is a transmembrane glycoprotein selectively expressed in high endothelial veins of mucosal lymphoid organs and in the intestinal lamina propria. Under inflammatory conditions, various cytokines promote high expression of MAdCAM-1 on endothelial cells, which then mediate the migration of α4β7-expressing leukocytes to the site of inflammation. Targeting either integrin α4β7 or MAdCAM-1 can reduce intestinal inflammation. Currently, there are no small molecule compounds specifically targeting α4β7-mediated inflammation on the market. Natalizumab, a humanized monoclonal antibody targeting the α4 subunit, is used clinically to treat multiple sclerosis and Crohn's disease, but has been associated with PML (progressive multifocal leukoencephalopathy) as a side effect. Therefore, there is a need to develop a small molecule compound that can inhibit the integrin α4β7 protein for the treatment of diseases associated with integrin α4β7 activity or expression. Summary of the Invention

[0005] The present invention develops a novel, highly effective, and safer integrin α4β7 inhibitor. These compounds have good pharmacokinetic properties (e.g., good AUC, bioavailability, low clearance, and good liver microsomal stability) and good safety, have good selectivity for α4β7, and exhibit better permeability in the Caco2 test. They are used to treat diseases associated with integrin α4β7, such as intestinal inflammatory diseases.

[0006] The present invention provides a compound of general formula (I) or general formula (II) or its stereoisomer, racemate, deuterated substance, solvate, prodrug, metabolite, pharmaceutically acceptable salt or cocrystal, wherein

[0007] In some embodiments, Selected from

[0008] In some embodiments, R 1 Selected from H or C1-4 alkyl;

[0009] In some embodiments, R 1 Selected from H, methyl or ethyl;

[0010] In some embodiments, R 2 Selected from halogen, OH, cyano, C 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 1-6 Alkoxy, C 3-6 Cycloalkyl or 4 to 7 membered heterocyclic group, wherein the alkyl, alkenyl, alkynyl, alkoxy, cycloalkyl or heterocyclic group is optionally substituted by 1 to 4 atoms selected from deuterium, halogen, CN, C 1-6 Alkyl or C 1-6 substituted by an alkoxy substituent;

[0011] In some embodiments, R 2 Selected from F, methyl, CHF2, CH2F, CF3;

[0012] In some embodiments, R 2a Each independently selected from halogen, OH, cyano, C 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 1-6 Alkoxy, C 3-6 Cycloalkyl or 4 to 7 membered heterocyclic group, wherein the alkyl, alkenyl, alkynyl, alkoxy, cycloalkyl or heterocyclic group is optionally substituted by 1 to 4 atoms selected from deuterium, halogen, CN, C 1-6 Alkyl or C 1-6 substituted by an alkoxy substituent;

[0013] In some embodiments, R 2a is selected from F, Cl, Br, OH, cyano, methyl, ethyl, isopropyl, methoxy, ethoxy or cyclopropyl, wherein the methyl, ethyl, isopropyl, methoxy, ethoxy or cyclopropyl is optionally substituted by 1 to 4 groups selected from deuterium, halogen, CN, C 1-4 Alkyl or C 1-4 substituted by an alkoxy substituent;

[0014] In some embodiments, Selected from

[0015] In some embodiments, Selected from

[0016] In some embodiments, Selected from

[0017] In some embodiments, R 3 Selected from

[0018] In some embodiments, R 3 Selected from

[0019] In some embodiments, R 3 Selected from

[0020] In some embodiments, R 3 Selected from

[0021] In some embodiments, R 3a 、R 3b are independently selected from H, halogen, cyano, C 1-6 Alkyl, C 1-6 Alkoxy, the alkyl or alkoxy is optionally substituted by 1 to 4 atoms selected from deuterium, halogen, CN, C 1-6 Alkyl or C 1-6 substituted by an alkoxy substituent;

[0022] In some embodiments, R 3a 、R 3b are each independently selected from H, F, Cl, Br, cyano, methyl, ethyl, isopropyl, methoxy or ethoxy, wherein the methyl, ethyl, isopropyl, methoxy or ethoxy is optionally substituted by 1 to 4 atoms selected from deuterium, halogen, CN, C 1-4 Alkyl or C 1-4 substituted by an alkoxy substituent;

[0023] In some embodiments, R 3a 、R 3b are each independently selected from H, F, Cl, Br, cyano, methyl, ethyl, isopropyl, methoxy or ethoxy, wherein the methyl, ethyl, isopropyl, methoxy or ethoxy group is optionally substituted with 1 to 4 substituents selected from deuterium, F, Cl, Br, CN, methyl, ethyl, propyl, isopropyl, methoxy or ethoxy;

[0024] In some embodiments, R 4 Selected from

[0025] In some embodiments, n is selected from 0, 1, 2, 3, or 4;

[0026] Optionally, the compound represented by general formula (I) is not the following compound and its stereoisomers

[0027] As a first embodiment of the present invention, the compound represented by the aforementioned general formula (I) or its stereoisomer, racemate, deuterated substance, solvate, prodrug, metabolite, pharmaceutically acceptable salt or cocrystal,

[0028] R 1 Selected from H or C 1-4 alkyl;

[0029] R 2 Selected from halogen, OH, cyano, C 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 1-6 Alkoxy, C 3-6 Cycloalkyl or 4 to 7 membered heterocyclic group, wherein the alkyl, alkenyl, alkynyl, alkoxy, cycloalkyl or heterocyclic group is optionally substituted by 1 to 4 atoms selected from deuterium, halogen, CN, C 1-6 Alkyl or C 1-6 substituted by an alkoxy substituent;

[0030] R 2a Each independently selected from halogen, OH, cyano, C 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 1-6 Alkoxy, C 3-6 Cycloalkyl or 4 to 7 membered heterocyclic group, wherein the alkyl, alkenyl, alkynyl, alkoxy, cycloalkyl or heterocyclic group is optionally substituted by 1 to 4 atoms selected from deuterium, halogen, CN, C 1-6 Alkyl or C 1-6 substituted by an alkoxy substituent;

[0031] R 3 Selected from

[0032] R 3a 、R 3b are independently selected from H, halogen, cyano, C 1-6 Alkyl, C 1-6 Alkoxy, the alkyl or alkoxy is optionally substituted by 1 to 4 atoms selected from deuterium, halogen, CN, C 1-6 Alkyl or C 1-6 substituted by an alkoxy substituent;

[0033] n is selected from 0, 1, 2, 3 or 4;

[0034] Provided that the compound represented by general formula (I) is not the following compound and its stereoisomers

[0035] As a second embodiment of the present invention, the compound represented by the aforementioned general formula (I) or its stereoisomer, racemate, deuterated substance, solvate, prodrug, metabolite, pharmaceutically acceptable salt or cocrystal,

[0036] Selected from

[0037] R 1 Selected from H, methyl or ethyl;

[0038] R 2 Selected from F, methyl, CHF2, CH2F, CF3;

[0039] R 2a Each is independently selected from F, Cl, Br, OH, cyano, methyl, ethyl, isopropyl, methoxy, ethoxy or cyclopropyl, wherein the methyl, ethyl, isopropyl, methoxy, ethoxy or cyclopropyl is optionally substituted by 1 to 4 groups selected from deuterium, halogen, CN, C 1-4 Alkyl or C 1-4 substituted by an alkoxy substituent;

[0040] R 3a 、R 3b are each independently selected from H, F, Cl, Br, cyano, methyl, ethyl, isopropyl, methoxy or ethoxy, wherein the methyl, ethyl, isopropyl, methoxy or ethoxy is optionally substituted by 1 to 4 atoms selected from deuterium, halogen, CN, C 1-4 Alkyl or C 1-4 substituted by an alkoxy substituent;

[0041] The definitions of the remaining substituents are consistent with those of the present invention.

[0042] As a third embodiment of the present invention, the compound represented by the aforementioned general formula (I) or its stereoisomer, racemate, deuterated substance, solvate, prodrug, metabolite, pharmaceutically acceptable salt or cocrystal,

[0043] R 3 Selected from

[0044] Selected from

[0045] The definitions of the remaining substituents are consistent with those in Scheme 1 or 2 of the present invention.

[0046] The present invention relates to some specific compounds of general formula (I), which are selected from Table E-1.

[0047] Table E-1

[0048] The present invention relates to a pharmaceutical composition comprising the compound of the present invention or its stereoisomer, racemate, deuterated substance, solvate, prodrug, metabolite, pharmaceutically acceptable salt or cocrystal, and a pharmaceutically acceptable carrier.

[0049] The present invention relates to the use of a compound of the present invention or its stereoisomer, racemate, deuterated substance, solvate, prodrug, metabolite, pharmaceutically acceptable salt or cocrystal in the preparation of a medicament for treating diseases related to α4β7 activity or expression, preferably in the preparation of a medicament for intestinal inflammatory diseases (preferably Crohn's disease or ulcerative colitis).

[0050] The present invention relates to a pharmaceutical composition or pharmaceutical formulation comprising a therapeutically effective amount of a compound of the present invention, or a stereoisomer, racemate, deuterated form, solvate, prodrug, metabolite, pharmaceutically acceptable salt, or cocrystal thereof, and a pharmaceutically acceptable excipient. The pharmaceutical composition may be in the form of a unit dosage form (the amount of the active ingredient in a unit dosage form is also referred to as the "drug strength").

[0051] The present invention also provides a method for treating a disease in a mammal, comprising administering to the mammal a therapeutically effective amount of a compound of the present invention, or a stereoisomer, racemate, deuterated form, solvate, prodrug, metabolite, pharmaceutically acceptable salt, cocrystal, or pharmaceutical composition thereof. In some embodiments, the mammal of the present invention comprises a human.

[0052] As used herein, an "effective amount" or "therapeutically effective amount" refers to the administration of a sufficient amount of a compound disclosed herein to alleviate to some extent one or more symptoms of the disease or condition being treated, such as an inflammatory bowel disease (preferably Crohn's disease or ulcerative colitis). In some embodiments, the result is a reduction and / or alleviation of the signs, symptoms, or causes of the disease, or any other desired change in a biological system. For example, an "effective amount" for therapeutic use is the amount of a composition comprising a compound disclosed herein required to provide a clinically significant reduction in disease symptoms.Examples of therapeutically effective amounts include, but are not limited to, 1-1500 mg, 1-1200 mg, 1-1000 mg, 1-900 mg, 1-800 mg, 1-700 mg, 1-600 mg, 2-600 mg, 3-600 mg, 4-600 mg, 5-600 mg, 6-600 mg, 10-600 mg, 20-600 mg, 25-600 mg, 30-600 mg, 40-600 mg, 50-600 mg, 60-600 mg, 70-600 mg, 75-600 mg, 80-600 mg, 90-600 mg, 100-600 mg, 200-600 mg, 1-500 mg, 2-500mg, 3-500mg, 4-500mg, 5-500mg, 6-500mg, 10-500mg, 20-500mg, 25-500mg, 30-500mg, 40-500mg, 50-500mg, 60-500mg, 70-500mg, 75-500mg , 80-500mg, 90-500mg, 100-500mg, 125-500mg, 150-500mg, 200-500mg, 250-500mg, 300-500mg, 400-500mg, 5-400mg, 10-400mg, 20-400mg, 25-40 0mg, 30-400mg, 40-400mg, 50-400mg, 60-400mg, 70-400mg, 75-400mg, 80-400mg, 90-400mg, 100-400mg, 125-400mg, 150-400mg, 200-400mg, 250- 400mg, 300-400mg, 1-300mg, 2-300mg, 5-300mg, 10-300mg, 20-300mg, 25-300mg, 30-300mg, 40-300mg, 50-300mg, 60-300mg, 70-300mg, 75-300mg , 80-300mg, 90-300mg, 100-300mg, 125-300mg, 150-300mg, 200-300mg, 250-300mg, 1-200mg, 2-200mg, 5-200mg, 10-200mg, 20-200mg, 25-200mg, 30-200mg, 40-200mg, 50-200mg, 60-200mg, 70-200mg, 75-200mg, 80-200mg, 90-200mg, 100-200mg, 125-200mg, 150-200mg, 80-1000mg, 80-800mg.

[0053] In some embodiments, the pharmaceutical composition includes but is not limited to 1-1000 mg, 20-800 mg, 40-800 mg, 40-400 mg, 25-200 mg, 1 mg, 5 mg, 10 mg, 15 mg, 20 mg, 25 mg, 30 mg, 35 mg, 40 mg, 45 mg, 50 mg, 55 mg, 65 mg, 70 mg, 75 mg, 80 mg, 85 mg, 90 mg, 95 mg, 100 mg, 110 mg, 120 mg, 1 25 mg, 130 mg, 140 mg, 150 mg, 160 mg, 170 mg, 180 mg, 190 mg, 200 mg, 210 mg, 220 mg, 230 mg, 240 mg, 250 mg, 300 mg, 320 mg, 400 mg, 480 mg, 500 mg, 600 mg, 640 mg, 840 mg of a compound of the present invention or a stereoisomer, racemate, deuterated substance, solvate, prodrug, metabolite, pharmaceutically acceptable salt or cocrystal thereof.

[0054] A method for treating a disease in a mammal, comprising administering to a subject a therapeutically effective amount of a compound of the present invention or a stereoisomer, racemate, deuterated form, solvate, prodrug, metabolite, pharmaceutically acceptable salt or cocrystal thereof, wherein the therapeutically effective amount is preferably 1-1500 mg, and the disease is preferably an intestinal inflammatory disease (preferably Crohn's disease or ulcerative colitis).

[0055] A method for treating a disease in a mammal, comprising administering to a subject a compound of the present invention or a stereoisomer, racemate, deuterated form, solvate, prodrug, metabolite, pharmaceutically acceptable salt or cocrystal thereof at a daily dose of 1-1000 mg / day, wherein the daily dose can be a single dose or divided doses. In some embodiments, the daily dose includes but is not limited to 10-1500 mg / day, 10-1000 mg / day, 10-800 mg / day, 25-800 mg / day, 50-800 mg / day, 100-800 mg / day, 200-800 mg / day, / day, 25-400 mg / day, 50-400 mg / day, 100-400 mg / day, 200-400 mg / day, in some embodiments, daily doses include but are not limited to 10 mg / day, 20 mg / day, 25 mg / day, 50 mg / day, 80 mg / day, 100 mg / day, 125 mg / day, 150 mg / day, 160 mg / day, 200 mg / day, 300 mg / day, 320 mg / day, 400 mg / day, 480 mg / day, 600 mg / day, 640 mg / day, 800 mg / day, 1000 mg / day.

[0056] The present invention relates to a kit, which may include a composition in single-dose or multi-dose form, wherein the kit contains a compound of the present invention or a stereoisomer, racemate, deuterated form, solvate, prodrug, metabolite, pharmaceutically acceptable salt or cocrystal thereof, and the amount of the compound of the present invention or its stereoisomer, racemate, deuterated form, solvate, prodrug, metabolite, pharmaceutically acceptable salt or cocrystal is the same as the amount in the above-mentioned pharmaceutical composition.

[0057] The amount of the compound of the invention or its stereoisomer, racemate, deuterated form, solvate, prodrug, metabolite, pharmaceutically acceptable salt or cocrystal in the present invention is in each case calculated as the free base.

[0058] "Preparation specifications" refers to the weight of the main drug contained in each vial, tablet or other unit preparation.

[0059] Unless stated otherwise, the terms used in the specification and claims have the following meanings.

[0060] The carbon, hydrogen, oxygen, sulfur, nitrogen or F, Cl, Br, I involved in the groups and compounds of the present invention include their isotopes, and the carbon, hydrogen, oxygen, sulfur or nitrogen involved in the groups and compounds of the present invention are optionally replaced by one or more of their corresponding isotopes, wherein the isotopes of carbon include 12 C. 13 C and 14 C, hydrogen isotopes include protium (H), deuterium (D, also called heavy hydrogen), tritium (T, also called super tritium), oxygen isotopes include 16 O. 17 O and 18 O, sulfur isotopes include 32 S. 33 S. 34 S and 36 S, nitrogen isotopes include 14 N and 15 N, fluorine isotopes include 17 F and 19 F, chlorine isotopes include 35 Cl and 37 Isotopes of Cl, bromine include 79 Br and 81 Br.

[0061] "Halogen" refers to F, Cl, Br or I.

[0062] "Halogen-substituted" refers to substitution with F, Cl, Br or I, including but not limited to substitution with 1 to 10 substituents selected from F, Cl, Br or I, substitution with 1 to 6 substituents selected from F, Cl, Br or I, and substitution with 1 to 4 substituents selected from F, Cl, Br or I. "Halogen-substituted" is abbreviated as "halo".

[0063] "Alkyl" refers to a substituted or unsubstituted straight-chain or branched saturated aliphatic hydrocarbon group, including but not limited to alkyl groups of 1 to 20 carbon atoms, alkyl groups of 1 to 8 carbon atoms, alkyl groups of 1 to 6 carbon atoms, and alkyl groups of 1 to 4 carbon atoms. Non-limiting examples include methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, neobutyl, tert-butyl, n-pentyl, isopentyl, neopentyl, n-hexyl, and various branched chain isomers thereof; alkyl groups appearing herein have the same definition as this one. Alkyl groups can be monovalent, divalent, trivalent, or tetravalent.

[0064] "Alkylene" refers to substituted or unsubstituted straight-chain and branched divalent saturated hydrocarbon groups, including -(CH2) v -(v is an integer from 1 to 10), examples of alkylene include but are not limited to methylene, ethylene, propylene and butylene.

[0065] "Cycloalkyl" refers to a substituted or unsubstituted saturated carbocyclic hydrocarbon radical, typically having 3 to 10 carbon atoms, non-limiting examples of which include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, or cycloheptyl. Cycloalkyl groups as used herein are as defined above. Cycloalkyl groups can be monovalent, divalent, trivalent, or tetravalent.

[0066] "Heterocycloalkyl" refers to a substituted or unsubstituted saturated cyclic hydrocarbon group containing heteroatoms, including but not limited to 3 to 10 atoms, 3 to 8 atoms, including 1 to 3 heteroatoms selected from N, O or S. The N and S optionally substituted in the ring of the heterocycloalkyl can be oxidized to various oxidation states. The heterocycloalkyl group can be connected to a heteroatom or a carbon atom, the heterocycloalkyl group can be connected to an aromatic ring or a non-aromatic ring, and the heterocycloalkyl group can be connected to a bridged ring or a spiro ring. Non-limiting examples include oxirane, aziridine, oxetanyl, azetidinyl, tetrahydrofuranyl, tetrahydro-2H-pyranyl, dioxolane, dioxane, pyrrolidinyl, piperidinyl, imidazolidinyl, oxazolidinyl, oxazinyl, morpholinyl, hexahydropyrimidinyl, piperazinyl. The heterocycloalkyl group can be monovalent, divalent, trivalent or tetravalent.

[0067] "Alkenyl" refers to substituted or unsubstituted straight and branched unsaturated hydrocarbon groups having at least one, typically one, two or three carbon-carbon double bonds, with a backbone of 2 to 10, 2 to 6 or 2 to 4 carbon atoms. Examples of alkenyl groups include, but are not limited to, vinyl, allyl, 1-propenyl, 2-propenyl, 1-butenyl, 2-butenyl, 3-butenyl, 1-pentenyl, 2-pentenyl, 3-pentenyl, 4-pentenyl, 1-methyl-1-butenyl, 2-methyl-1-butenyl, 2- Methyl-3-butenyl, 1-hexenyl, 2-hexenyl, 3-hexenyl, 4-hexenyl, 5-hexenyl, 1-methyl-1-pentenyl, 2-methyl-1-pentenyl, 1-heptenyl, 2-heptenyl, 3-heptenyl, 4-heptenyl, 1-octenyl, 3-octenyl, 1-nonenyl, 3-nonenyl, 1-decenyl, 4-decenyl, 1,3-butadiene, 1,3-pentadiene, 1,4-pentadiene, and 1,4-hexadiene; alkenyl groups appearing herein have the same definition as this one. Alkenyl groups may be monovalent, divalent, trivalent, or tetravalent.

[0068] "Alkynyl" refers to substituted or unsubstituted straight and branched unsaturated hydrocarbon groups having at least one, typically one, two or three carbon-carbon triple bonds, with a backbone comprising 2 to 10 carbon atoms, including but not limited to 2 to 6 carbon atoms in the backbone, and 2 to 4 carbon atoms in the backbone. Examples of alkynyl groups include but are not limited to ethynyl, propargyl, 1-propynyl, 2-propynyl, 1-butynyl, 2-butynyl, 3-butynyl, 1-pentynyl, 2-pentynyl, 3-pentynyl, 4-pentynyl, 5-pentynyl, 6-pentynyl, 7-pentynyl, 8-pentynyl, 9-pentynyl, 10-pentynyl, 11-pentynyl, 12-pentynyl, 13-pentynyl, 14-pentynyl, 15-pentynyl, 16-pentynyl, 17-pentynyl, 18-pentynyl, 19-pentynyl, 20-pentynyl, 21-pentynyl, 22-pentynyl, 23-pentynyl, 24-pentynyl, 25-pentynyl, 26-pentynyl, 27-pentynyl, 28-pentynyl, 29-pentynyl, 30-pentynyl, 31-pentynyl, 32-pentynyl, 33-pentynyl, 34-pentynyl, 35-pentynyl, 36-pentynyl, 37-pentynyl, 38-pentynyl, 39-pentynyl, 40-pentynyl, 41-pentynyl, 42-pentynyl, 43-pentynyl, 44-pentynyl, 45-pentynyl, 46-pentynyl, 47-pentynyl, 48-pentyn Alkynyl, 1-methyl-1-butynyl, 2-methyl-1-butynyl, 2-methyl-3-butynyl, 1-hexynyl, 2-hexynyl, 3-hexynyl, 4-hexynyl, 5-hexynyl, 1-methyl-1-pentynyl, 2-methyl-1-pentynyl, 1-heptynyl, 2-heptynyl, 3-heptynyl, 4-heptynyl, 1-octynyl, 3-octynyl, 1-nonynyl, 3-nonynyl, 1-decynyl, 4-decynyl, and the like; an alkynyl group may be monovalent, divalent, trivalent, or tetravalent.

[0069] "Alkoxy" refers to a substituted or unsubstituted -O-alkyl group. Non-limiting examples include methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, sec-butoxy, tert-butoxy, n-pentoxy, n-hexoxy, cyclopropyloxy, and cyclobutyloxy.

[0070] "1 to X substituents substituted" means substituted by 11, 2, 3 ... X substituents, X is selected from any integer between 1 and 10. For example, "0 to 4 substituents substituted" means substituted by 0, 1, 2, 3 or 4 substituents. For example, "1 to 4 R k "Substituted" means replaced by 1, 2, 3 or 4 R kFor example, "the heterobridged ring is optionally substituted by 0 to 4 substituents selected from H or F" means that the heterobridged ring is optionally substituted by 0, 1, 2, 3 or 4 substituents selected from H or F.

[0071] XY-membered rings (3≤X<Y, Y is any integer between 4 and 12) include rings with X+1, X+2, X+3, X+4, ..., Y members. Rings include heterocycles, carbocycles, aromatic rings, aryl groups, heteroaryl groups, cycloalkyl groups, heteromonocycles, heteroplexes, heterospirocycles, or heterobridged rings. For example, "4-7-membered heteromonocycle" refers to a 4-, 5-, 6-, or 7-membered heteromonocycle, and "5-10-membered heteroplexes" refers to a 5-, 6-, 7-, 8-, 9-, or 10-membered heteroplex.

[0072] "Optional" or "optionally" means that the subsequently described event or circumstance may but need not occur, and the description includes instances where the event or circumstance occurs and instances where it does not. For example, "alkyl optionally substituted with F" means that the alkyl group may but need not be substituted with F, and the description includes instances where the alkyl group is substituted with F and instances where the alkyl group is not substituted with F.

[0073] "Pharmaceutically acceptable salt" or "pharmaceutically acceptable salt thereof" refers to a salt of the compound of the present invention that retains the biological effectiveness and properties of the free acid or free base, and the free acid is obtained by reacting with a non-toxic inorganic base or organic base, or the free base is obtained by reacting with a non-toxic inorganic acid or organic acid.

[0074] "Pharmaceutical composition" refers to a mixture of one or more compounds of the present invention, their pharmaceutically acceptable salts or prodrugs and other chemical components, wherein "other chemical components" refers to pharmaceutically acceptable carriers, excipients and / or one or more other therapeutic agents.

[0075] "Carrier" refers to a material that does not cause significant irritation to an organism and does not abrogate the biological activity and properties of the administered compound.

[0076] "Animal" is meant to include mammals, such as humans, companion animals, zoo animals, and livestock, preferably humans, horses, or dogs.

[0077] "Stereoisomers" refer to isomers resulting from different spatial arrangements of atoms in a molecule, including cis-trans isomers, enantiomers, and conformational isomers. DETAILED DESCRIPTION

[0078] In order to accomplish the purpose of the present invention, the compounds used in the reactions described herein are prepared according to organic synthesis techniques known to those skilled in the art, starting from commercially available chemicals and / or compounds described in the chemical literature. "Commercially available chemicals" are obtained from standard commercial sources, including Shanghai Aladdin Biochemical Technology Co., Ltd., Shanghai McLean Biochemical Technology Co., Ltd., Sigma-Aldrich, Alfa Aesar (China) Chemical Co., Ltd., Tokyo Chemical Industry Development Co., Ltd., Anage Chemical, Shanghai Titan Technology Co., Ltd., Kelon Chemical, Bailingwei Technology Co., Ltd., etc.

[0079] The following examples illustrate the technical solutions of the present invention in detail, but the protection scope of the present invention includes but is not limited to them.

[0080] The structures of the compounds were confirmed by nuclear magnetic resonance (NMR) and / or mass spectrometry (MS). NMR shifts (δ) are given in units of 10-6 (ppm). NMR measurements were performed using a Bruker Avance III 400 and Bruker Avance 300 NMR spectrometer. The solvents used were deuterated dimethyl sulfoxide (DMSO-d6), deuterated chloroform (CDCl3), and deuterated methanol (CD3OD). The internal standard was tetramethylsilane (TMS).

[0081] MS was determined using (Agilent 6120B (ESI) and Agilent 6120B (APCI));

[0082] HPLC determination was performed using an Agilent 1260DAD high-pressure liquid chromatograph (Zorbax SB-C18 100 × 4.6 mm, 3.5 μM);

[0083] Thin layer chromatography silica gel plates used were Yantai Huanghai HSGF254 or Qingdao GF254 silica gel plates. The specifications of the silica gel plates used for thin layer chromatography (TLC) were 0.15 mm to 0.20 mm, and the specifications used for thin layer chromatography separation and purification products were 0.4 mm to 0.5 mm.

[0084] Column chromatography generally uses Yantai Huanghai silica gel 200-300 mesh silica gel as the carrier.

[0085] Retention time (Rt, retention time): Unless otherwise specified in the examples, it is the retention time of the analytical method, and the method is as follows:

[0086] Detection wavelength: 254nm / 210nm Flow rate: 1.0ml / min Column temperature: 35℃

[0087] Injection volume: 2 μl Acquisition time: 10 min

[0088] Gradient elution program:

[0089] Acidic conditions: Mobile phase A is 0.05% TFA solution, mobile phase B is acetonitrile

[0090] Chromatographic column brand: Yuexu, chromatographic column model: Xtimate C18 4.6*50mm, 3μm

[0091] The retention time of HPLC or SFC indicates the order in which the compounds appear.

[0092] Synthesis of intermediate 1a:

[0093] Step 1: Synthesis of 1a-2

[0094] Dissolve 1a-1 (1720 mg, 6.96 mmol, synthesized according to US20150148347) in trifluoroacetic acid (25 mL). Add triethylsilane (25 mL). React at room temperature for 48 h. Then add triethylsilane (15 mL) and stir overnight. After concentration under reduced pressure, the product was purified by column chromatography on a silica gel B-1 column to afford crude 1a-2 (1700 mg).

[0095] 1 H NMR (400MHz, CDCl3) δ6.61(t,1H),2.99–2.84(m,4H),2.15–2.02(m,2H).

[0096] Step 2: Synthesis of 1a-3

[0097] Under nitrogen, 1a-2 (1700 mg, 7.29 mmol) was dissolved in dry THF (70 mL), cooled to -78°C, and lithium diisopropylamide (1.56 g, 14.58 mmol) was slowly added. The mixture was stirred at -78°C for 0.5 h, and dry DMF (2.6 g, 36.45 mmol) was added. Stirring was continued at -78°C for 1 h. The reaction was quenched by the addition of ammonium chloride solution (50 mL). The mixture was extracted with ethyl acetate (80 mL × 3), dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and purified by silica gel column chromatography to afford 1a-3 (1.58 g, two-step yield: 83.02%).

[0098] Step 3: Synthesis of 1a-4

[0099] Under nitrogen protection, 1a-3 (1580 mg, 6.05 mmol) and R-tert-butylsulfenamide (1100 mg, 9.07 mmol) were dissolved in tetrahydrofuran (35 mL), and tetraethyl titanate (2070 mg, 9.07 mmol) was slowly added. The reaction was carried out at 45 ° C for 15 h, and the mixture was concentrated under reduced pressure to obtain a crude product. The crude product was purified by silica gel flash column chromatography to obtain 1a-4 (2100 mg, yield 95.29%).

[0100] LC-Ms m / z(ESI):364.2[M+H] + .

[0101] Step 4: Synthesis of 1a-5

[0102] Zinc powder (1.2 g, 18.34 mmol) was added to dry tetrahydrofuran (8 mL), and the nitrogen atmosphere was replaced three times. CuCl (389 mg, 3.93 mmol) was added, and the reaction was carried out at 60°C for 2 h. After cooling to room temperature, ethyl bromoacetate (1.09 g, 6.55 mmol) was slowly added, and the reaction was carried out at 60°C for 1 h. After cooling to 0°C, a solution of 1a-4 (478 mg, 1.31 mmol) in tetrahydrofuran (3 mL) was added, and the mixture was stirred at 0°C for 3 h. After filtration, saturated ammonium chloride solution (30 mL) was added, and the mixture was extracted with ethyl acetate (40 ml x 3). The mixture was washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. After flash column chromatography, 1a-5 (498 mg, 84.04% yield) was obtained.

[0103] LC-Ms m / z(ESI):452.4[M+H] + .

[0104] Step 5: Synthesis of 1a-6

[0105] 1a-5 (448 mg, 0.99 mmol) was dissolved in dichloromethane (3 mL), 4N hydrochloric acid in dioxane (2.5 mL) was added, and the mixture was stirred at room temperature for 3 h. The mixture was concentrated under reduced pressure to obtain the hydrochloride salt of crude 1a-6.

[0106] LC-Ms m / z(ESI):348.3[M+H] +

[0107] Step 6: Synthesis of 1a

[0108] The hydrochloride salt of the crude product 1a-6 from the previous step was dissolved in tetrahydrofuran (4 mL) and water (4 mL). Sodium carbonate (210 mg, 1.98 mmol) and Boc2O (240 mg, 1.09 mmol) were added and stirred at room temperature for 3 h. Water (10 mL) was added and the mixture was extracted with ethyl acetate (20 mL x 3). The mixture was dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and purified by flash column chromatography to afford 1a (420 mg, 86.31% yield over two steps, ee >99%; chiral HPLC retention time: 6.535 min). HPLC conditions for chiral analysis: Instrument: Shimadzu LC-20A; Chiral Column: CHIRALCEL AD-H, 4.6 x 250 mm, 5 μm; Mobile Phase: n-hexane-ethanol (90:10); Flow Rate: 1 mL / min; Column Temperature: 35°C; Detection Wavelength: 210 nm; Injection Volume: 10 μL; Run Time: 20 min.

[0109] LC-Ms m / z(ESI):350.0[M+H-Boc] + .

[0110] 1 H NMR (400MHz, CD3OD) δ5.54–5.27(m,1H),4.16–4.00(m,2H),3.09–2.88(m,5H),2.86–2.74(m,1H),2.24–2.08(m,2H),1.39(s,9H),1.18(t,3H).

[0111] Example 1: Preparation of Compound 1

[0112] Step 1: Synthesis of 1b

[0113] Under nitrogen, 1a (500 mg, 1.12 mmol) was dissolved in 1,4-dioxane (20 mL) and water (2 mL). 2,4,6-Trimethylphenylboronic acid (220 mg, 1.34 mmol), XPhos Pd G2 (180 mg, 0.22 mmol), and potassium phosphate (710 mg, 3.36 mmol) were added sequentially, and the mixture was reacted at 100°C for 24 h. Ethyl acetate (80 mL) was added for extraction, and the mixture was washed with water (50 mL × 2) and saturated brine (50 mL × 1), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure and purified by silica gel column chromatography to afford 1b (260 mg, yield: 47.81%).

[0114] LC-Ms m / z(ESI):488.5[M+H] +

[0115] Step 2: Synthesis of 1c

[0116] 1b (260 mg, 0.53 mmol) was dissolved in dichloromethane (5 mL), and 1,4-dioxane hydrochloride (5 mL) was added and reacted for 1 h. The solvent was removed under reduced pressure to obtain 200 mg of the crude trifluoroacetic acid salt of compound 1c.

[0117] Step 3: Synthesis of 1e

[0118] Under nitrogen, intermediate 1d (190 mg, 0.52 mmol) was dissolved in dry DMF (5 mL), and HATU (400 mg, 1.04 mmol) and DIPEA (270 mg, 2.08 mmol) were added sequentially. After stirring at room temperature for 40 min, 1c (200 mg, 0.52 mmol) was added. The reaction was allowed to proceed overnight at room temperature, and ethyl acetate (80 mL) was added. The mixture was washed sequentially with water (20 mL × 2) and saturated brine (20 mL × 1), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure and purified by silica gel column chromatography to afford 1e (180 mg, yield: 47.78%).

[0119] LC-Ms m / z(ESI):730.7[M+H] +

[0120] Step 4: Synthesis of Compound 1-1 and Compound 1-2

[0121] Compound 1e (180 mg, 0.25 mmol) was dissolved in 6 mL of tetrahydrofuran and 2 mL of water, and lithium hydroxide monohydrate (15 mg, 0.63 mmol) was added. The mixture was allowed to react at room temperature for 30 min. The pH was adjusted to 5-6 with 1N hydrochloric acid, and the mixture was concentrated under reduced pressure to afford crude compound 1. This crude product was then separated and purified by prep-HPLC using a Waters 2767 preparative column: SunFire@Prep C18 (19 mm × 150 mm); mobile phase composition: mobile phase A: acetonitrile, mobile phase B: water (containing 0.1% ammonium acetate) to afford compound 1-1 (15 mg, analytical HPLC retention time = 4.259 min, yield 8.67%) and compound 1-2 (20 mg, analytical HPLC retention time = 4.353 min, yield 11.56%).

[0122] Compound 1-1:

[0123] LC-Ms m / z(ESI):702.7[M+H] +

[0124] 1H NMR(400MHz,CD3OD)δ7.77(s,1H),6.92(s,2H),6.88(s,1H),5.76–5.69(m,1H),5. 67–5.60(m,1H),4.06–3.94(m,4H),3.37–3.23(m,2H),2.99–2.92(m,2H),2.91–2. 81(m,3H),2.68–2.60(m,1H),2.49–2.37(m,4H),2.29(s,3H),2.10–2.00(m,2H),2 .00–1.94(m,2H),1.92(s,3H),1.87(s,3H),1.46–1.33(m,1H),0.98–0.88(m,6H).

[0125] Compound 1-2:

[0126] LC-Ms m / z(ESI):702.7[M+H] +

[0127] 1 H NMR(400MHz,CD3OD)δ7.66(s,1H),6.96–6.89(m,3H),5.89(dd,1H),5.60(t,1H),4.16–4.05(m,4H),3.46–3.31(m,2H),3.03–2.88(m,3H),2.8 6–2.70(m,2H),2.53–2.40(m,5H),2.30(s,3H),2.13–2.02(m,2H),1.95 –1.88(m,7H),1.78–1.66(m,1H),1.43–1.32(m,1H),0.94–0.82(m,6H).

[0128] Example 2: Preparation of Compound 2

[0129] Step 1: Synthesis of 2a

[0130] Under nitrogen, 1a (400 mg, 0.89 mmol) was dissolved in 1,4-dioxane (6 mL) and water (0.6 mL). 2,6-Dimethyl-4-methoxyphenylboronic acid (240 mg, 1.33 mmol), XPhos Pd G2 (CAS: 1310584-14-5) (110 mg, 0.13 mmol), and potassium phosphate (570 mg, 2.67 mmol) were added sequentially. The mixture was reacted at 100°C for 24 h. Ethyl acetate (80 mL) was added for extraction, and the mixture was washed with water (50 mL × 2) and saturated brine (50 mL × 1), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure and purified by silica gel column chromatography to afford 2a (230 mg, yield: 51.32%).

[0131] LC-Ms m / z(ESI):404.5[M+H-Boc] +

[0132] Step 2: Synthesis of 2b

[0133] 2a (230 mg, 0.46 mmol) was dissolved in dichloromethane (6 mL), and hydrochloric acid-1,4-dioxane (6 mL) was added and reacted for 1 h. The solvent was removed under reduced pressure to obtain the hydrochloride salt of crude product 2b.

[0134] Step 3: Synthesis of 2c

[0135] Under nitrogen, 1d (190 mg, 0.50 mmol) was dissolved in dry DMF (10 mL), and EDCI (CAS: 7084-11-9) (190 mg, 1.0 mmol), HOBt (CAS: 2592-95-2) (140 mg, 1.0 mmol), and DIPEA (260 mg, 2.0 mmol) were added sequentially. After stirring at room temperature for 40 min, 2b (200 mg, 0.50 mmol) was added. The reaction was allowed to proceed overnight at room temperature, and ethyl acetate (80 mL) was added. The mixture was washed sequentially with water (20 mL × 2) and saturated brine (20 mL × 1), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure and purified by silica gel column chromatography to afford 2c (260 mg, yield: 68.67%).

[0136] LC-Ms m / z(ESI):764.7[M+H] +

[0137] Step 4: Synthesis of Compound 2-1 and Compound 2-2

[0138] 2c (260 mg, 0.34 mmol) was dissolved in 9 mL of tetrahydrofuran and 3 mL of water, and lithium hydroxide monohydrate (20 mg, 0.85 mmol) was added. The reaction was allowed to react at room temperature for 30 min. The pH was adjusted to 5-6 with 1N hydrochloric acid, and the mixture was concentrated under reduced pressure to afford crude compound 2. This crude product was then separated and purified by prep-HPLC using a Waters 2767 preparative column: SunFire@Prep C18 (19 mm × 150 mm); (mobile phase composition: mobile phase A: acetonitrile, mobile phase B: water (containing 0.1% ammonium acetate)) to afford compound 2-1 (20 mg, retention time = 4.098 min, yield 7.99%) and compound 2-2 (20 mg, retention time = 4.190 min, yield 9.98%).

[0139] Compound 2-1:

[0140] LC-Ms m / z(ESI):736.7[M+H] +

[0141] 1 H NMR(400MHz,CD3OD)δ7.78(s,1H),6.80(s,1H),6.66(s,2H),5.69(t,2H),5.27– 5.05(m,1H),3.94–3.82(m,2H),3.78(s,3H),3.58–3.40(m,2H),3.04–2.84(m,5H ),2.81–2.73(m,1H),2.73–2.59(m,2H),2.48–2.39(m,2H),2.11–1.99(m,2H),1. 99–1.93(m,2H),1.91(s,3H),1.83(s,3H),1.48–1.35(m,1H),1.00–0.87(m,6H).

[0142] Compound 2-2:

[0143] LC-Ms m / z(ESI):736.7[M+H] +

[0144] 1H NMR(400MHz,CD3OD)δ7.71(s,1H),6.89(s,1H),6.70(s,2H),5.85–5.77(m,1H),5.69–5.6 1(m,1H),5.36–5.14(m,1H),4.24–4.10(m,2H),3.89–3.69(m,5H),3.20–3.11(m,2H),3.02 –2.94(m,2H),2.88–2.71(m,3H),2.66–2.57(m,1H),2.53–2.44(m,2H),2.13–2.02(m,2H) ,1.95(s,6H),1.92–1.84(m,1H),1.83–1.73(m,1H),1.42–1.27(m,1H),0.94–0.83(m,6H).

[0145] Example 3: Preparation of Compound 3

[0146] Referring to the synthetic route and preparation method of compound 2, a crude product of compound 3 was obtained, and the crude product was separated and purified by prep-HPLC (instrument: Waters 2767 preparative chromatography column: SunFire@Prep C18 (19 mm × 150 mm); (mobile phase composition: mobile phase A: acetonitrile, mobile phase B: water (containing 0.1% ammonium acetate) to obtain compound 3-1 (10 mg, yield 6.92%, HPLC retention time: 4.116 min) and compound 3-2 (10 mg, yield 6.92%, HPLC retention time: 4.186 min).

[0147] Compound 3-1: LC-Ms m / z (ESI): 750.4 [M+H] +

[0148] 1 H NMR (400MHz, CD3OD) δ7.85(s,1H),6.80(s,1H),6.69–6.63(m,2H),5.74–5,64(m,2H),5.33–5.13(m,1H),3.78(s,3H),3.26–3.01(m, 3H),3.00–2.73(m,9H),2.48–2.39(m,2H),2.33–1.99(m,4H),1.98–1.88(m,5H),1.82(s,3H),1.48–1.35(m,1H),1.00–0.88(m,6H).

[0149] Compound 3-2: LC-Ms m / z (ESI): 750.4 [M+H] +

[0150] 1 H NMR(400MHz,CD3OD)δ7.86(s,1H),6.87(s,1H),6.69(s,2H),5.78–5.72(m,1H),5.70–5.62(m,1H), 5.36–5.17(m,1H),3.79(s,3H),3.32–3.29(m,1H),3.28–3.21(m,2H),3.18–3.02(m,3H),3.01–2.94 (m,2H),2.93–2.82(m,3H),2.70–2.61(m,1H),2.52–2.44(m,2H),2.38–2.13(m,2H),2.12–2.02(m, 2H),1.97–1.92(m,6H),1.92–1.83(m,1H),1.81–1.71(m,1H),1.40–1.27(m,1H),0.93–0.81(m,6H).

[0151] Example 4: Preparation of Compound 4

[0152] Referring to the synthetic route and preparation method of compound 2, a crude product of compound 4 was obtained, and the crude product was separated and purified by prep-HPLC (instrument: Waters 2767 preparative chromatography column: SunFire@Prep C18 (19 mm × 150 mm); (mobile phase composition: mobile phase A: acetonitrile, mobile phase B: water (containing 0.1% ammonium acetate) to obtain compound 4-1 (11 mg, yield 11.42%, HPLC retention time: 4.195 min) and compound 4-2 (12 mg, yield 12.46%, HPLC retention time: 4.262 min).

[0153] LC-Ms m / z (ESI) of compound 4-1: 738.7 [M+H] +

[0154] 1H NMR (400 MHz, CD3OD) δ7.85 (s, 1H), 6.88–6.81 (m, 2H), 6.79 (s, 1H), 5.74–5.64 (m, 2H), 5.34–5.13 (m, 1H), 3.26–3.02 (m, 3H), 3.02–2.74 (m, 9H), 2.48–2.40 (m, 2H), 2.36–2.00 (m, 4H), 2.00–1.91 (m, 5H), 1.86 (s, 3H), 1.47–1.36 (m, 1H), 0.99–0.89 (m, 6H). LC-Ms m / z (ESI) of compound 4-2: 738.7 [M+H] +

[0155] 1 H NMR(400MHz,CD3OD)δ7.86(s,1H),6.91–6.82(m,3H),5.78–5.71(m,1H),5.70–5.61(m,1H),5. 36–5.17(m,1H),3.33–3.28(m,1H),3.28–3.20(m,2H),3.19–3.02(m,3H),3.02–2.94(m,2H),2 .93–2.82(m,3H),2.72–2.60(m,1H),2.52–2.44(m,2H),2.40–2.14(m,2H),2.14–2.03(m,2H), 2.01–1.93(m,6H),1.92–1.83(m,1H),1.82–1.71(m,1H),1.41–1.27(m,1H),0.94–0.82(m,6H).

[0156] Example 5: Preparation of Compound 5

[0157] Referring to the synthetic route and preparation method of compound 2, a crude product of compound 5 was obtained, and the crude product was separated and purified by prep-HPLC (instrument: Waters 2767 preparative chromatography column: SunFire@Prep C18 (19 mm × 150 mm); (mobile phase composition: mobile phase A: acetonitrile, mobile phase B: water (containing 0.1% ammonium acetate) to obtain compound 5-1 (10 mg, yield 6.92%, HPLC retention time: 4.184 min) and compound 5-2 (10 mg, yield 6.92%, HPLC retention time: 4.261 min).

[0158] LC-Ms m / z (ESI) of compound 5-1: 750.4 [M+H] +

[0159] 1 H NMR(400MHz,CD3OD)δ7.87(s,1H),6.80(s,1H),6.69–6.61(m,2H),5.74–5,64(m,2H),5.33–5.13(m,1H),3.78(s,3H),3.24 –2.71(m,12H),2.48–2.40(m,2H),2.34–2.00(m,4H),1.98–1.87(m,5H),1.82(s,3H),1.48–1.35(m,1H),0.99–0.89(m,6H).

[0160] LC-Ms m / z (ESI) of compound 5-2: 750.4 [M+H] +

[0161] 1 H NMR(400MHz,CD3OD)δ7.83(s,1H),6.87(s,1H),6.69(s,2H),5.78–5.71(m,1H),5.70 –5.63(m,1H),5.39–5.18(m,1H),3.79(s,3H),3.43–3.33(m,1H),3.27–3.01(m,5H),3 .01–2.94(m,2H),2.93–2.80(m,3H),2.70–2.61(m,1H),2.52–2.44(m,2H),2.37–2.0 2(m,4H),1.97–1.90(m,6H),1.90–1.71(m,2H),1.41–1.26(m,1H),0.96–0.82(m,6H).

[0162] Example 6: Preparation of Compound 6

[0163] Referring to the synthetic route and preparation method of compound 2, a crude product of compound 6 was obtained, and the crude product was separated and purified by prep-HPLC (instrument: Waters 2767 preparative chromatography column: SunFire@Prep C18 (19 mm × 150 mm); (mobile phase composition: mobile phase A: acetonitrile, mobile phase B: water (containing 0.1% ammonium acetate) to obtain compound 6-1 (12 mg, yield 10.38%, HPLC retention time: 4.267 min) and compound 6-2 (14 mg, yield 12.11%, HPLC retention time: 4.357 min).

[0164] Compound 6-1: LC-Ms m / z (ESI): 738.7 [M+H] +

[0165] 1 H NMR (400MHz, CD3OD) δ7.87(s,1H),6.88–6.81(m,2H),6.80(s,1H),5.73–5.63(m,2H),5.34–5.13(m,1H),3.24–3.12(m,2H),3. 11–2.72(m,10H),2.50–2.39(m,2H),2.35–2.00(m,4H),1.99–1.89(m,5H),1.86(s,3H),1.48–1.35(m,1H),0.99–0.89(m,6H).

[0166] Compound 6-2: LC-Ms m / z (ESI): 738.7 [M+H] +

[0167] 1 H NMR(400MHz,CD3OD)δ7.83(s,1H),6.92–6.83(m,3H),5.78–5.70(m,1H),5. 70–5.62(m,1H),5.39–5.19(m,1H),3.43–3.34(m,1H),3.25–2.95(m,7H),2 .93–2.81(m,3H),2.70–2.62(m,1H),2.52–2.44(m,2H),2.40–2.03(m,4H), 2.01–1.95(m,6H),1.92–1.71(m,2H),1.41–1.26(m,1H),0.93–0.82(m,6H).

[0168] Example 7: Preparation of Compound 7

[0169] Intermediate 7a was synthesized with reference to WO2021076890 and the synthetic route and preparation method of compound 2 to obtain a crude product of compound 7. The crude product was separated and purified by prep-HPLC to obtain the trifluoroacetate salt of compound 7-1 (11 mg, HPLC retention time: 3.905 min) and the trifluoroacetate salt of compound 7-2 (12 mg, HPLC retention time: 4.114 min).

[0170] HPLC preparation conditions: Instrument: Waters 2767; Preparative chromatographic column: SunFire@Prep C18 (19 mm × 150 mm); Mobile phase composition: Mobile phase A: acetonitrile, Mobile phase B: Water (containing 0.1% trifluoroacetic acid)

[0171] Trifluoroacetate of compound 7-1: LC-Ms m / z (ESI): 682.3 [M+H] +

[0172] 1 H NMR(400MHz,CD3OD)δ7.55(s,1H),6.67(s,2H),6.37(s,1H),5.79–5.67(m,2H),4.68–4.21(m,4H),3.78(s,3H),3.52–3.40(m,2H) ,3.14–3.03(m,1H),2.98–2.70(m,5H),2.49–2.41(m,2H),2.23(s,3H),2.10–1.81(m,10H),1.43–1.31(m,2H),0.96–0.91(m,6H).

[0173] Trifluoroacetate of compound 7-2: LC-Ms m / z (ESI): 682.4 [M+H] +

[0174] 1 H NMR (400MHz, CD3OD) δ7.57(s,1H),6.70(s,2H),6.43(s,1H),5.79–5.66(m,2H),4.66–4.30(m,4H),3.79(s,3H),3.52–3.46(m,2H),3.08–2.94 (m,3H),2.92–2.70(m,3H),2.49(m,2H),2.26(s,3H),2.13–2.02(m,2H) ,1.95(s,6H),1.80–1.67(m,2H),1.33–1.23(m,2H),0.91–0.83(m,6H).

[0175] Example 8: Preparation of Compound 8

[0176] Step 1: Synthesis of 8b

[0177] 8a (3.0 g, 8.64 mmol) was dissolved in 30 mL of ultra-dry DCE. 3-Azetidine 3-acetonitrile (1.06 g, 12.96 mmol) and AcOH (0.10 mg, 1.73 mmol) were added sequentially. The reaction was allowed to proceed at room temperature for 1 h. Sodium triacetoxyborohydride (3.66 g, 17.28 mmol) was added and the reaction continued for 16 h. The mixture was then extracted with 200 mL of ethyl acetate and 100 mL of saturated aqueous NaHCO₃. The aqueous phase was then extracted with ethyl acetate (100 mL x 1). The ethyl acetate layers were combined, washed with saturated brine (100 mL x 1), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by column chromatography to afford 8b (2.8 g, 78.41% yield).

[0178] LC-Ms m / z(ESI):414.0[M+H] +

[0179] Step 2: Synthesis of 8c

[0180] 8c (2.8 g, 6.78 mmol) was dissolved in THF (30 mL) and water (10 mL), lithium hydroxide (331.55 mg, 13.85 mmol) was added, and the reaction was continued for 30 min. 8c was purified by reverse phase chromatography to obtain 8c (2.0 g, yield: 76.80%).

[0181] LC-Ms m / z(ESI):385.2[M+H] +

[0182] Step 3: 8D synthesis

[0183] Under nitrogen, 8c (323.40 mg, 0.84 mmol) and 4a (328.44 mg, 0.84 mmol) were dissolved in dry DMF (5 mL). HOBt (227.00 mg, 1.68 mmol), EDCI (322.06 mg, 1.68 mmol), and DIPEA (434.25 mg, 3.36 mmol) were added sequentially. The mixture was allowed to react at room temperature overnight. Ethyl acetate (100 mL) was added, and the mixture was washed sequentially with water (40 mL × 2) and saturated brine (40 mL × 1), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. 8d (400.0 mg, yield: 62.82%) was obtained by silica gel column chromatography.

[0184] LC-Ms m / z(ESI):758.3[M+H] +

[0185] Step 4: Synthesis of Compound 8-1 and Compound 8-2

[0186] 8d (400.0 mg, 0.53 mmol) was dissolved in 3 mL of methanol and 1 mL of water, and lithium hydroxide monohydrate (27.0 mg, 1.12 mmol) was added. The mixture was allowed to react at room temperature for 30 min. The pH was adjusted to 5-6 with 1N hydrochloric acid, and the mixture was concentrated under reduced pressure to afford crude compound 9. This crude product was then separated and purified by SFC to afford compound 8-1 (50 mg, yield 12.92%, SFC retention time: 0.949 min) and compound 8-2 (50 mg, SFC retention time: 1.185 min).

[0187] SFC preparation conditions: instrument: Waters 150 Prep-SFC; chromatographic column: Chiral IK column; mobile phase: A is CO2; B is a methanol solution containing 0.1% ammonia; elution conditions: 20% B isocratic elution; flow rate: 100 mL / min; pressure: 100 bar; column temperature: room temperature; detection wavelength: 220 nm.

[0188] Compound 8-1: LC-Ms m / z (ESI): 731.3 [M+H] +

[0189] 1 H NMR(400MHz,CD3OD)δ7.81(s,1H),6.88(d,2H),6.83(s,1H),5.75–5.67(m,2H),3.77–3.68(m,2H),3.59–3.46(m,3H),3.03–2.93(m,3H),2.88 –2.73(m,3H),2.66–2.58(m,2H),2.53–2.46(m,2H),2.14–2.04(m,2H) ,1.98(s,6H),1.84–1.77(m,2H),1.39–1.27(m,2H),0.94–0.83(m,6H).

[0190] Compound 8-2: LC-Ms m / z (ESI): 731.3 [M+H] +

[0191] 1 H NMR (400MHz, CD3OD) δ7.85(s,1H),6.90–6.78(m,3H),5.82–5.67(m,2H),4.45–4.27(m,4H),3.99–3.85(m,1H),3.40–3.31(m,2H),3.16–3. 04(m,1H),3.00–2.91(m,2H),2.91–2.74(m,3H),2.51–2.36(m,2H),2 .11–1.90(m,7H),1.87(s,3H),1.47–1.36(m,1H),1.02–0.90(m,6H).

[0192] Example 9: Preparation of Compound 9

[0193] Referring to the synthetic route and method of compound 8, compound 9-1 (50 mg, SFC retention time: 1.181 min) and compound 9-2 (50 mg, SFC retention time: 1.708 min) were prepared by SFC.

[0194] SFC preparation conditions: instrument: Waters 150 Prep-SFC; chromatographic column: Chiral IK column; mobile phase: A is CO2; B is a methanol solution containing 0.1% ammonia; elution conditions: 35% B isocratic elution; flow rate: 100 mL / min; pressure: 100 bar; column temperature: room temperature; detection wavelength: 220 nm.

[0195] Compound 9-1: LC-Ms m / z (ESI): 743.3 [M+H] +

[0196] 1 H NMR (400MHz, CD3OD) δ7.82(s,1H),6.84(s,1H),6.70(s,2H),5.76–5.68(m,2H),3.86–3.76(m,5H),3.69–3.51(m,3H),3.04–2.94(m,3H),2.91 –2.78(m,3H),2.69–2.61(m,2H),2.53–2.45(m,2H),2.12–2.03(m,2H) ,1.95(s,6H),1.85–1.77(m,2H),1.37–1.26(m,1H),0.94–0.84(m,6H).

[0197] Compound 9-2: LC-Ms m / z (ESI): 743.3 [M+H] +

[0198] 1 H NMR(400MHz,CD3OD)δ7.80(s,1H),6.77(s,1H),6.69–6.62(m,2H),5.75–5.65(m,2H),3. 78(s,3H),3.66–3.54(m,2H),3.47–3.34(m,3H),3.10–3.00(m,1H),2.97–2.89(m,2H),2 .89–2.80(m,1H),2.71–2.58(m,2H),2.58–2.49(m,2H),2.49–2.39(m,2H),2.09–1.98(m ,2H),1.98–1.93(m,2H),1.89(s,3H),1.81(s,3H),1.49–1.37(m,1H),1.00–0.91(m,6H).

[0199] Example 10: Preparation of Compound 10

[0200] Referring to the synthetic route and method of compound 8, compound 10-1 (50 mg, yield 12.92%, SFC retention time: 0.999 min) and compound 10-2 (50 mg, yield 12.92%, SFC retention time: 1.548 min) were prepared by SFC.

[0201] SFC preparation conditions: instrument: Waters 150 Prep-SFC; chromatographic column: Chiral OD column; mobile phase: A is CO2; B is methanol solution; elution conditions: 25% B isocratic elution; flow rate: 100 mL / min; pressure: 100 bar; column temperature: room temperature; detection wavelength: 220 nm.

[0202] Compound 10-1: LC-Ms m / z (ESI): 731.3 [M+H] +

[0203] 1 H NMR(400MHz,CD3OD)δ7.81(s,1H),7.15–7.09(m,1H),7.02–6.95(m,1H),6.83( s,1H),5.76–5.67(m,2H),3.72–3.64(m,2H),3.53–3.41(m,3H),3.03–2.94(m, 3H),2.88–2.68(m,3H),2.64–2.57(m,2H),2.53–2.45(m,2H),2.15–2.05(m,2H ),1.98–1.88(m,6H),1.85–1.78(m,2H),1.39–1.26(m,1H),0.94–0.83(m,6H).

[0204] Compound 10-2: LC-Ms m / z (ESI): 731.3 [M+H] +

[0205] 1H NMR(400MHz,CD3OD)δ7.80(s,1H),7.11–7.04(m,1H),7.00–6.91(m,1H),6.76( s,1H),5.74–5.65(m,2H),3.64–3.53(m,2H),3.46–3.31(m,3H),3.10–3.01(m, 1H),3.00–2.91(m,2H),2.90–2.81(m,1H),2.71–2.39(m,6H),2.10–2.01(m,2H ),1.99–1.91(m,2H),1.87–1.77(m,6H),1.49–1.38(m,1H),1.00–0.91(m,6H).

[0206] Example 11: Preparation of Compound 11

[0207] Referring to the synthetic route and method of compound 8, compound 11-1 (45 mg, yield 11.80%, HPLC retention time: 4.302 min) and compound 11-2 (45 mg, yield 11.80%, HPLC retention time: 4.406 min) were prepared by pre-HPLC.

[0208] HPLC preparation conditions: Instrument: Waters 2767; Preparative Column: SunFire@Prep C18 (19 mm × 150 mm); Mobile phase composition: Mobile phase A: acetonitrile, Mobile phase B: Water (containing 0.1% ammonium acetate)

[0209] Compound 11-1: LC-Ms m / z (ESI): 720.3 [M+H] +

[0210] 1 H NMR(400MHz,CD3OD)δ7.78(s,1H),6.89–6.81(m,3H),5.76–5.69(m,1H),5.6 8–5.60(m,1H),4.14–4.03(m,2H),3.67–3.56(m,2H),3.33–3.20(m,2H),3.00 –2.81(m,6H),2.67–2.57(m,1H),2.51–2.42(m,2H),2.13–2.02(m,2H),2.01– 1.94(m,5H),1.92(s,3H),1.46–1.33(m,1H),1.23(d,3H),0.98–0.88(m,6H).

[0211] Compound 11-2: LC-Ms m / z (ESI): 720.3 [M+H] +

[0212] 1 H NMR(400MHz,CD3OD)δ7.67(s,1H),6.94–6.84(m,3H),5.91–5.84(m,1H),5.63– 5.56(m,1H),4.25–4.13(m,2H),3.78–3.68(m,2H),3.34–3.34(m,2H),3.04–2. 88(m,4H),2.87–2.71(m,2H),2.53–2.42(m,3H),2.15–2.04(m,2H),2.00–1.89 (m,7H),1.77–1.65(m,1H),1.44–1.31(m,1H),1.25(d,3H),0.94–0.84(m,6H).

[0213] Example 12: Preparation of Compound 12

[0214] Referring to the synthetic route and method of compound 8, pre-HPLC obtained compound 12-1 (44 mg, yield 11.35%, HPLC retention time: 4.235 min) and compound 12-2 (44 mg, yield 11.35%, HPLC retention time: 4.334 min).

[0215] HPLC preparation conditions: Instrument: Waters 2767; Preparative Column: SunFire@Prep C18 (19 mm × 150 mm); Mobile phase composition: Mobile phase A: acetonitrile, Mobile phase B: Water (containing 0.1% ammonium acetate)

[0216] Compound 12-1: LC-Ms m / z (ESI): 732.3 [M+H] +

[0217] 1H NMR(400MHz,CD3OD)δ7.66(s,1H),6.92(s,1H),6.69(s,2H),5.88(dd,1H),5.59( t,1H),4.25–4.13(m,2H),3.79(s,3H),3.76–3.66(m,2H),3.42–3.33(m,2H),3.0 2–2.88(m,4H),2.85–2.71(m,2H),2.53–2.42(m,3H),2.13–2.02(m,2H),2.00–1. 90(m,7H),1.76–1.66(m,1H),1.44–1.33(m,1H),1.25(d,3H),0.93–0.85(m,6H).

[0218] Compound 12-2: LC-Ms m / z (ESI): 732.3 [M+H] +

[0219] 1 H NMR(400MHz,CD3OD)δ7.77(s,1H),6.88(s,1H),6.68(s,2H),5.76–5.69(m,1H ),5.66–5.59(m,1H),4.12–4.01(m,2H),3.78(s,3H),3.65–3.54(m,2H),3.30 –3.21(m,2H),2.99–2.82(m,6H),2.67–2.58(m,1H),2.51–2.43(m,2H),2.10– 1.91(m,7H),1.89(s,3H),1.46–1.35(m,1H),1.23(d,3H),0.97–0.89(m,6H).

[0220] Example 13: Preparation of Compound 13

[0221] Referring to the synthetic route and method of compound 8, compound 13-1 (44 mg, yield 11.35%, HPLC retention time: 4.289 min) and compound 13-2 (44 mg, yield 11.35%, HPLC retention time: 4.378 min) were obtained by preparative HPLC purification.

[0222] HPLC preparation conditions: Instrument: Waters 2767; Preparative Column: SunFire@Prep C18 (19 mm × 150 mm); Mobile phase composition: Mobile phase A: acetonitrile, Mobile phase B: Water (containing 0.1% ammonium acetate)

[0223] Compound 13-1: LC-Ms m / z (ESI): 720.3 [M+H] +

[0224] 1 H NMR(400MHz,CD3OD)δ7.78(s,1H),7.14–7.07(m,1H),7.01–6.93(m,1H),6.87(s, 1H),5.76–5.69(m,1H),5.68–5.60(m,1H),4.14–4.02(m,2H),3.66–3.55(m,2H),3 .35–3.19(m,2H),3.01–2.81(m,6H),2.69–2.59(m,1H),2.52–2.40(m,2H),2.13–2 .03(m,2H),2.01–1.81(m,8H),1.46–1.34(m,1H),1.23(d,3H),0.98–0.89(m,6H).

[0225] Compound 13-2: LC-Ms m / z (ESI): 720.3 [M+H] +

[0226] 1 H NMR(400MHz,CD3OD)δ7.67(s,1H),7.16–7.09(m,1H),7.02–6.95(m,1H),6.92(s,1H), 5.92–5.83(m,1H),5.63–5.56(m,1H),4.25–4.13(m,2H),3.77–3.66(m,2H),3.43–3.33 (m,2H),3.04–2.71(m,6H),2.53–2.44(m,3H),2.15–2.04(m,2H),2.00–1.92(m,4H),1. 92–1.86(m,3H),1.76–1.67(m,1H),1.44–1.31(m,1H),1.25(d,3H),0.93–0.84(m,6H).

[0227] Example 14: Preparation of Compound 14

[0228] Referring to the synthetic route and method of compound 8, compound 14-1 (45 mg, yield 11.80%, HPLC retention time: 4.289 min) and compound 14-2 (45 mg, yield 11.80%, HPLC retention time: 4.378 min) were obtained by preparative HPLC purification.

[0229] HPLC preparation conditions: Instrument: Waters 2767; Preparative Column: SunFire@Prep C18 (19 mm × 150 mm); Mobile phase composition: Mobile phase A: acetonitrile, Mobile phase B: Water (containing 0.1% ammonium acetate)

[0230] Compound 14-1: LC-Ms m / z (ESI): 732.3 [M+H] +

[0231] 1 H NMR(400MHz,CD3OD)δ7.77(s,1H),7.06(d,1H),6.90–6.81(m,2H),5.76–5.68(m, 1H),5.67–5.60(m,1H),4.13–4.00(m,2H),3.82(s,3H),3.65–3.55(m,2H),3.36–3 .21(m,1H),2.99–2.81(m,6H),2.68–2.59(m,1H),2.52–2.36(m,2H),2.12–1.91(m ,4H),1.90–1.75(m,6H),1.47–1.33(m,1H),1.26–1.18(m,3H),0.98–0.88(m,6H).

[0232] Compound 14-2: LC-Ms m / z (ESI): 732.3 [M+H] +

[0233] 1 H NMR(400MHz,CD3OD)δ7.67(s,1H),7.08(d,1H),6.91(s,1H),6.86(d,1H),5.92–5.84(m ,1H),5.63–5.55(m,1H),4.26–4.13(m,2H),3.83(s,3H),3.77–3.68(m,2H),3.43–3.34 (m,2H),3.03–2.87(m,4H),2.86–2.71(m,2H),2.51–2.42(m,3H),2.13–2.02(m,2H),2. 00–1.80(m,7H),1.77–1.65(m,1H),1.45–1.32(m,1H),1.25(d,3H),0.93–0.85(m,6H).

[0234] Example 15: Preparation of Compound 15

[0235] Referring to the synthetic route and method of compound 8, SFC obtained compound 15-1 (46 mg, yield 11.71%, SFC retention time: 1.11 min) and compound 15-2 (46 mg, yield 11.71%, SFC retention time: 1.30 min).

[0236] SFC preparation conditions:

[0237] Instrument: Waters 150 Prep-SFC; chromatographic column: Chiral OD column; mobile phase: A is CO2; B is methanol solution; elution conditions: 15% B isocratic elution; flow rate: 120 mL / min; pressure: 100 bar; column temperature: room temperature; detection wavelength: 220 nm.

[0238] Compound 15-1: LC-Ms m / z (ESI): 742.3 [M+H] +

[0239] 1 H NMR (400MHz, CD3OD) δ7.83(s,1H),6.88(d,2H),6.82(s,1H),5.76–5.65(m,2H),3.74–3.58(m,4H),3.05–2.95(m,3H),2.91–2.73(m, 3H),2.70–2.57(m,2H),2.53–2.46(m,2H),2.16–2.04(m,2H),1.98(s,6H),1.88–1.73(m,2H),1.40–1.26(m,1H),0.93–0.82(m,6H).

[0240] Compound 15-2: LC-Ms m / z (ESI): 742.3 [M+H] +

[0241] 1 H NMR(400MHz,CD3OD)δ7.80(s,1H),6.87–6.80(m,2H),6.77(s,1H),5.75–5.64(m,2H),3.70–3.54(m,4H),3.10–3.01(m,1H),2.99–2.90(m,2H),2 .89–2.81(m,1H),2.81–2.68(m,2H),2.67–2.49(m,2H),2.47–2.39(m,2H ),2.10–1.88(m,7H),1.85(s,3H),1.50–1.36(m,1H),0.99–0.88(m,6H).

[0242] Example 16: Preparation of Compound 16

[0243] Referring to the synthetic route and method of compound 8, compound 16-1 (46 mg, yield 11.71%, HPLC retention time: 4.389 min) and compound 16-2 (46 mg, yield 11.71%, HPLC retention time: 4.470 min) were obtained by preparative HPLC purification.

[0244] HPLC preparation conditions: Instrument: Waters 2767; Preparative Column: SunFire@Prep C18 (19 mm × 150 mm); Mobile phase composition: Mobile phase A: acetonitrile, Mobile phase B: Water (containing 0.1% ammonium acetate)

[0245] Compound 16-1: LC-Ms m / z (ESI): 742.3 [M+H] +

[0246] 1 H NMR (400MHz, CD3OD) δ7.80(s,1H),7.12–7.04(m,1H),7.00–6.92(m,1H),6.77(s,1H),5.76–5.63(m,2H),3.71–3.53(m,4H),3.10–3.01(m, 1H),2.99–2.90(m,2H),2.90–2.82(m,1H),2.80–2.50(m,4H),2.48–2 .40(m,2H),2.10–1.73(m,10H),1.48–1.37(m,1H),1.00–0.89(m,6H).

[0247] Compound 16-2: LC-Ms m / z (ESI): 742.3 [M+H] +

[0248] 1 H NMR(400MHz,CD3OD)δ7.83(s,1H),7.16–7.08(m,1H),7.02–6.94(m,1H),6 .82(s,1H),5.77–5.65(m,2H),3.74–3.56(m,4H),3.06–2.95(m,3H),2.92 –2.72(m,3H),2.71–2.57(m,2H),2.52–2.43(m,2H),2.15–2.03(m,2H),1. 98–1.87(m,6H),1.87–1.72(m,2H),1.39–1.27(m,1H),0.95–0.81(m,6H).

[0249] Example 17: Preparation of Compound 17

[0250] Referring to the synthetic route and method of compound 8, compound 17-1 (40 mg, yield 10.03%, SFC retention time: 1.147 min) and compound 17-2 (40 mg, SFC retention time: 1.599 min) were prepared by SFC.

[0251] SFC preparation conditions: instrument: Waters 150 Prep-SFC; chromatographic column: Chiral OD column; mobile phase: A is CO2; B is methanol solution; elution conditions: 25% B isocratic elution; flow rate: 120 mL / min; pressure: 100 bar; column temperature: room temperature; detection wavelength: 220 nm.

[0252] Compound 17-1: LC-Ms m / z (ESI): 754.3 [M+H] +

[0253] 1 H NMR (400MHz, CD3OD) δ7.83(s,1H),6.82(s,1H),6.69(s,2H),5.77–5.65(m,2H),3.79(s,3H),3.74–3.58(m,4H),3.04–2.94(m,3H),2.90–2 .72(m,3H),2.70–2.56(m,2H),2.52–2.46(m,2H),2.12–2.02(m,2H), 1.95(s,6H),1.87–1.73(m,2H),1.38–1.26(m,1H),0.93–0.83(m,6H).

[0254] Compound 17-2: LC-Ms m / z (ESI): 754.3 [M+H] +

[0255] 1 H NMR(400MHz,CD3OD)δ7.80(s,1H),6.77(s,1H),6.68–6.62(m,2H),5.76–5.64(m,2H),3.78(s,3H),3.71–3.53(m,4H),3 .10–3.01(m,1H),2.99–2.40(m,9H),2.09–1.91(m,4H),1.89(s,3H),1.81(s,3H),1.48–1.35(m,1H),1.00–0.88(m,6H).

[0256] Example 18: Preparation of Compound 18

[0257] Referring to the synthetic route and method of compound 2, compound 18-1 (40 mg, yield 10.32%, HPLC retention time: 4.500 min) and compound 18-2 (40 mg, yield 10.32%, HPLC retention time: 4.594 min) were obtained by preparative HPLC purification.

[0258] HPLC preparation conditions: instrument: Waters 2767 preparative chromatography column: SunFire@Prep C18 (19 mm×150 mm); mobile phase composition: mobile phase A: acetonitrile, mobile phase B: water (containing 0.1% ammonium acetate).

[0259] Compound 18-1: LC-Ms m / z (ESI): 732.3 [M+H] +

[0260] 1 H NMR (400MHz, CD3OD) δ7.77(s,1H),6.88(s,1H),6.66(s,2H),5.74–5.68(m,1H),5.67–5.60(m,1H),4.08–2.92(m,6H),3.36–3.22(m,2H),3 .00–2.82(m,5H),2.67–2.58(m,1H),2.51–2.35(m,4H),2.11–1.94(m ,4H),1.92(s,3H),1.88(s,3H),1.46–1.33(m,4H),0.99–0.88(m,6H).

[0261] Compound 18-2: LC-Ms m / z (ESI): 732.3 [M+H] +

[0262] 1 H NMR (400MHz, CD3OD) δ7.66(s,1H),6.92(s,1H),6.68(s,2H),5.93–5.85(m,1H),5.63–5.56(m,1H),4.15–3.98(m,6H),3.45–3.31(m,2H),3.03–2 .87(m,3H),2.86–2.71(m,2H),2.53–2.38(m,5H),2.13–2.03(m,2H),2.0 0–1.87(m,7H),1.78–1.68(m,1H),1.43–1.31(m,4H),0.94–0.84(m,6H).

[0263] Example 19: Preparation of Compound 19

[0264] Referring to the synthetic route and method of compound 2, compound 19-1 (40 mg, yield 10.49%, HPLC retention time: 4.244 min) and compound 19-2 (40 mg, yield 10.49%, HPLC retention time: 4.314 min) were obtained by preparative HPLC purification.

[0265] HPLC preparation conditions: Instrument: Waters 2767; Preparative Column: SunFire@Prep C18 (19 mm × 150 mm); Mobile phase composition: Mobile phase A: acetonitrile, Mobile phase B: Water (containing 0.1% ammonium acetate)

[0266] Compound 19-1: LC-Ms m / z (ESI): 720.3 [M+H] +

[0267] 1 H NMR(400MHz,CD3OD)δ7.85(s,1H),6.84(s,1H),6.65(s,2H),5.72–5.63(m,2H),4.07–3.96(m,2H),3.12–2.99(m,2H),2.97–2.86 (m,5H),2.75–2.62(m,7H),2.49–2.42(m,2H),2.09–1.93(m,4H),1.91(s,3H),1.85(s,3H),1.46–1.33(m,4H),0.98–0.87(m,6H).

[0268] Compound 19-2: LC-Ms m / z (ESI): 720.3 [M+H] +

[0269] 1 H NMR(400MHz,CD3OD)δ7.79(s,1H),6.91(s,1H),6.68(s,2H),5.86–5.77(m,1H),5.66–5.57(m,1H),4.08–3.98(m,2H),3.28–3.10(m,2H),3.02–2 .94(m,4H),2.83–2.72(m,7H),2.57–2.44(m,3H),2.13–2.01(m,2H),1.9 8–1.88(m,7H),1.80–1.69(m,1H),1.43–1.29(m,4H),0.94–0.82(m,6H).

[0270] Example 20: Preparation of Compound 20

[0271] Referring to the synthetic route and preparation method of compound 2, a crude product of compound 20 was obtained. The crude product was separated and purified by prep-HPLC to obtain the trifluoroacetate salt of compound 20-1 (102 mg, yield 21%, HPLC retention time: 3.978 min) and the trifluoroacetate salt of compound 20-2 (68 mg, yield 14%, HPLC retention time: 4.180 min).

[0272] HPLC preparation conditions: Instrument: Waters 2767; Preparative chromatographic column: SunFire@Prep C18 (19 mm × 150 mm); Mobile phase composition: Mobile phase A: acetonitrile, Mobile phase B: Water (containing 0.1% trifluoroacetic acid)

[0273] Trifluoroacetate salt of compound 20-1: LC-Ms m / z (ESI): 670.3 [M+H] +

[0274] 1 H NMR (400MHz, CD3OD) δ7.55(s,1H),6.85(m,2H),6.37(s,1H),5.79–5.66(m,2H),4.65–4.35(m,4H),3.52–3.42(m,2H),3. 14–3.03(m,1H),2.98–2.71(m,5H),2.45(m,2H),2.23(s,3H),2.11–1.82(m,10H),1.48–1.28(m,2H),0.97–0.90(m,6H).

[0275] Trifluoroacetate of compound 20-2: LC-Ms m / z (ESI): 670.3 [M+H] +

[0276] 1 H NMR (400MHz, CD3OD) δ7.57(s,1H),6.88(m,2H),6.43(s,1H),5.78–5.66(m,2H),4.68–4.17(m,4H),3.52–3.44(m,2H),3.09–2.94(m,3H),2 .93–2.70(m,3H),2.53–2.46(m,2H),2.26(s,3H),2.14–2.04(m,2H), 1.98(s,6H),1.84–1.67(m,2H),1.36–1.22(m,2H),0.91–0.82(m,6H).

[0277] Example 22: Preparation of Compound 22

[0278] Referring to the synthetic route and preparation method of compound 2, a crude product of compound 22 was obtained. The crude product was separated and purified by prep-HPLC to obtain the trifluoroacetate salt of compound 22-1 (87 mg, yield 31%, HPLC retention time: 4.300 min) and the trifluoroacetate salt of compound 22-2 (86 mg, yield 31%, HPLC retention time: 4.508 min).

[0279] HPLC preparation conditions: Instrument: Waters 2767; Preparative chromatographic column: SunFire@Prep C18 (19 mm × 150 mm); Mobile phase composition: Mobile phase A: acetonitrile, Mobile phase B: Water (containing 0.1% trifluoroacetic acid)

[0280] Trifluoroacetate of compound 22-1: LC-Ms m / z (ESI): 670.4 [M+H] +

[0281] 1 H NMR (400MHz, CD3OD) δ7.55(s,1H),7.10(m,1H),6.96(m,1H),6.37(s,1H),5.78–5.66(m,2H),4.71–4.25(m,4H),3.50–3.43(m,2H),3.14–3 .03(m,1H),2.99–2.71(m,5H),2.48–2.41(m,2H),2.23(s,3H),2.12– 1.93(m,3H),1.91–1.75(m,7H),1.42–1.26(m,2H),0.97–0.90(m,6H).

[0282] Trifluoroacetate of compound 22-2: LC-Ms m / z (ESI): 670.4 [M+H] +

[0283] 1H NMR(400MHz,CD3OD)δ7.58(s,1H),7.17–7.09(m,1H),6.99(m,1H),6.43(s,1H) ,5.79–5.66(m,2H),4.65–4.32(m,4H),3.52–3.44(m,2H),3.09–2.95(m,3H),2 .95–2.66(m,3H),2.53–2.45(m,2H),2.26(s,3H),2.15–2.05(m,2H),1.95(s,3 H),1.92–1.87(m,3H),1.85–1.67(m,2H),1.32–1.22(m,2H),0.91–0.83(m,6H).

[0284] Example 23: Preparation of Compound 23

[0285] Referring to the synthetic route and preparation method of compound 2, a crude product of compound 23 was obtained. The crude product was separated and purified by prep-HPLC to give the trifluoroacetate salt of compound 23-1 (92 mg, yield 33%, HPLC retention time: 4.479 min) and the trifluoroacetate salt of compound 23-2 (91 mg, yield 33%, HPLC retention time: 4.266 min).

[0286] HPLC preparation conditions: Instrument: Waters 2767; Preparative chromatographic column: SunFire@Prep C18 (19 mm × 150 mm); Mobile phase composition: Mobile phase A: acetonitrile, Mobile phase B: Water (containing 0.1% trifluoroacetic acid)

[0287] Trifluoroacetate salt of compound 23-1: LC-Ms m / z (ESI): 682.3 [M+H] +

[0288] 1 H NMR (400MHz, CD3OD) δ7.55(s,1H),7.05(m,1H),6.84(m,1H),6.37(s,1H),5.79–5.66(m,2H),4.65–4.30(m,4H),3.82(s,3H),3.50–3.40 (m,2H),3.14–3.03(m,1H),2.99–2.68(m,5H),2.47–2.36(m,2H),2.23(s,3H),2.09–1.70(m,10H),1.43–1.27(m,2H),0.96–0.90(m,6H).

[0289] Trifluoroacetate of compound 23-2: LC-Ms m / z (ESI): 682.3 [M+H] +

[0290] 1 H NMR (400MHz, CD3OD) δ7.57(s,1H),7.08(m,1H),6.87(m,1H),6.43(s,J=1.2Hz,1H ),5.79–5.66(m,2H),4.65–4.30(m,4H),3.83(s,3H),3.52–3.44(m,2H),3.08–2.9 4(m,3H),2.92–2.69(m,3H),2.50–2.42(m,2H),2.26(s,3H),2.12–2.02(m,2H),1 .90(s,3H),1.83(s,3H),1.79–1.68(m,2H),1.35–1.22(m,2H),0.93–0.82(m,6H).

[0291] Example 24: Preparation of Compound 24

[0292] Referring to the synthetic route and preparation method of compound 2, a crude product of compound 24 was obtained. The crude product was separated and purified by prep-HPLC to give the trifluoroacetate salt of compound 24-1 (53 mg, yield 19%, HPLC retention time: 3.928 min) and the trifluoroacetate salt of compound 24-2 (53 mg, yield 19%, HPLC retention time: 4.144 min).

[0293] HPLC preparation conditions: Instrument: Waters 2767; Preparative chromatographic column: SunFire@Prep C18 (19 mm × 150 mm); Mobile phase composition: Mobile phase A: acetonitrile, Mobile phase B: Water (containing 0.1% trifluoroacetic acid)

[0294] Trifluoroacetate salt of compound 24-1: LC-Ms m / z (ESI): 666.4 [M+H] +

[0295] 1H NMR(400MHz,CD3OD)δ7.61(s,1H),6.67(s,2H),6.37(s,1H),5.75–5.67(m,1H) ,5.60–5.53(m,1H),3.78(s,3H),3.31–3.18(m,2H),3.14–3.03(m,1H),2.99–2. 81(m,11H),2.48–2.38(m,2H),2.24(m,3H),2.20–1.99(m,3H),1.96–1.80(m,7 H),1.62–1.52(m,1H),1.27–1.15(m,1H),1.11–1.00(m,1H),0.93–0.86(m,6H).

[0296] Trifluoroacetate salt of compound 24-2: LC-Ms m / z (ESI): 666.4 [M+H] +

[0297] 1 H NMR(400MHz,CD3OD)δ7.64(s,1H),6.69(s,2H),6.43(s,1H),5.74–5.65(m ,1H),5.58–5.51(m,1H),3.79(s,3H),3.30–3.23(m,2H),3.08–2.94(m,9H ),2.93–2.83(m,3H),2.53–2.45(m,2H),2.27(m,3H),2.12–1.88(m,9H),1 .82–1.69(m,1H),1.56–1.44(m,1H),1.09–0.98(m,2H),0.84–0.76(m,6H).

[0298] Example 25: Preparation of Compound 25

[0299] Referring to the synthetic route and preparation method of compound 2, a crude product of compound 25 was obtained. The crude product was separated and purified by prep-HPLC to give the trifluoroacetate salt of compound 25-1 (87 mg, yield 32%, HPLC retention time: 4.307 min) and the trifluoroacetate salt of compound 25-2 (73 mg, yield 27%, HPLC retention time: 4.516 min).

[0300] HPLC preparation conditions: Instrument: Waters 2767; Preparative chromatographic column: SunFire@Prep C18 (19 mm × 150 mm); Mobile phase composition: Mobile phase A: acetonitrile, Mobile phase B: Water (containing 0.1% trifluoroacetic acid)

[0301] Trifluoroacetate salt of compound 25-1: LC-Ms m / z (ESI): 654.3 [M+H] +

[0302] 1 H NMR(400MHz,CD3OD)δ7.61(s,1H),7.14–7.06(m,1H),7.00–6.93(m,1H),6.37(s,1H),5.74 –5.66(m,1H),5.60–5.52(m,1H),3.29–3.19(m,2H),3.13–3.03(m,1H),2.96(s,8H),2.91– 2.84 (m, 3H), 2.48–2.41 (m, 2H), 2.24 (s, 3H), 2.20–2.00 (m, 3H), 1.95–1.76 (m, 7H), 1.65–1.51 (m, 1H), 1.27–1.16 (m, 1H), 1.11–1.00 (m, 1H), 0.93–0.87 (m, 6H). Trifluoroacetic acid salt of compound 25-2: LC-Ms m / z (ESI): 654.3 [M+H] +

[0303] 1 H NMR(400MHz,CD3OD)δ7.64(s,1H),7.16–7.09(m,1H),7.01–6.95(m,1H),6.43 (s,1H),5.73–5.65(m,1H),5.58–5.51(m,1H),3.30–3.24(m,2H),3.07–2.87( m,12H),2.53–2.45(m,2H),2.27(s,3H),2.14–2.04(m,2H),2.03–1.86(m,7H) ,1.81–1.70(m,1H),1.55–1.43(m,1H),1.08–1.00(m,2H),0.84–0.76(m,6H).

[0304] Example 26: Preparation of Compound 26

[0305] Referring to the synthetic route and preparation method of compound 2, a crude product of compound 26 was obtained. The crude product was separated and purified by prep-HPLC to give the trifluoroacetate salt of compound 26-1 (158 mg, yield 42%, HPLC retention time: 4.280 min) and the trifluoroacetate salt of compound 26-2 (139 mg, yield 37%, HPLC retention time: 4.492 min).

[0306] HPLC preparation conditions: Instrument: Waters 2767; Preparative chromatographic column: SunFire@Prep C18 (19 mm × 150 mm); Mobile phase composition: Mobile phase A: acetonitrile, Mobile phase B: Water (containing 0.1% trifluoroacetic acid)

[0307] Trifluoroacetate salt of compound 26-1: LC-Ms m / z (ESI): 666.4 [M+H] +

[0308] 1 H NMR(400MHz,CD3OD)δ7.61(s,1H),7.08–7.02(m,1H),6.86–6.80(m,1H),6.37(s,1H),5.7 6–5.66(m,1H),5.60–5.52(m,1H),3.82(s,3H),3.30–3.17(m,3H),3.13–3.02(m,1H),3.00 –2.91(m,7H),2.91–2.82(m,3H),2.46–2.38(m,2H),2.25(s,3H),2.20–2.00(m,3H),1.95 –1.70(m,7H),1.63–1.52(m,1H),1.25–1.15(m,1H),1.11–1.00(m,1H),0.92–0.86(m,6H).

[0309] Trifluoroacetate of compound 26-2: LC-Ms m / z (ESI): 666.4 [M+H] +

[0310] 1 H NMR(400MHz,CD3OD)δ7.64(s,1H),7.10–7.05(m,1H),6.88–6.84(m,1H),6.43(s,1H) ,5.73–5.66(m,1H),5.59–5.52(m,1H),3.83(s,3H),3.30–3.24(m,2H),3.07–2.84(m, 12H),2.50–2.43(m,2H),2.27(s,3H),2.12–2.04(m,2H),2.02–1.92(m,1H),1.91–1.8 1(m,6H),1.80–1.70(m,1H),1.54–1.44(m,1H),1.08–0.98(m,2H),0.83–0.77(m,6H).

[0311] Example 27: Preparation of Compound 27

[0312] Referring to the synthetic route and preparation method of compound 2, a crude product of compound 27 was obtained, and the crude product was separated and purified by prep-HPLC (instrument: Waters 2767 preparative chromatography column: SunFire@Prep C18 (19 mm × 150 mm); (mobile phase composition: mobile phase A: acetonitrile, mobile phase B: water (containing 0.1% ammonium acetate) to obtain compound 27-1 (157 mg, yield 42%, HPLC retention time: 4.385 min) and compound 27-2 (65 mg, yield 19%, HPLC retention time: 4.504 min).

[0313] Compound 27-1: LC-Ms m / z (ESI): 688.3 [M+H] +

[0314] 1 H NMR (400MHz, CD3OD) δ7.84–7.69(m,2H),6.95–6.59(m,3H),5.78–5.72(m,1H),5.65–5.59(m,1H),3.79(s,3H),3.40–3.33(m,1H),3.28–3.17( m,1H),3.01–2.64(m,11H),2.52–2.39(m,3H),2.13–2.02(m,2H),2.00 –1.90(m,7H),1.82–1.74(m,1H),1.44–1.32(m,1H),0.93–0.85(m,6H).

[0315] Compound 27-2: LC-Ms m / z (ESI): 688.3 [M+H] +

[0316] 1 H NMR (400MHz, CD3OD) δ7.87–7.68(m,2H),6.90–6.59(m,3H),5.67–5.61(m,1H),5.50–5.44(m,1H),3.78(s,3H),3.35–3.28(m,1H),3.20–3. 10(m,1H),2.98–2.81(m,4H),2.73(s,6H),2.64–2.60(m,2H),2.49–2 .42(m,2H),2.09–1.87(m,10H),1.45–1.35(m,1H),0.95–0.86(m,6H).

[0317] Example 28: Preparation of Compound 28

[0318] Referring to the synthetic route and preparation method of compound 2, a crude compound 28 was obtained, which was separated and purified by prep-HPLC (instrument: Waters 2767 preparative chromatography column: SunFire@Prep C18 (19 mm × 150 mm); (mobile phase composition: mobile phase A: acetonitrile, mobile phase B: water (containing 0.1% ammonium acetate) to obtain compound 28-1 (40 mg, yield 21%, HPLC retention time: 4.111 min) and compound 28-2 (21 mg, yield 12%, HPLC retention time: 4.233 min).

[0319] Compound 28-1: LC-Ms m / z (ESI): 688.3 [M+H] +

[0320] 1 H NMR(400MHz,CD3OD)δ7.85–7.68(m,2H),7.08–7.03(m,1H),6.90–6.59(m, 2H),5.68–5.58(m,1H),5.50–5.43(m,1H),3.82(s,3H),3.18–3.10(m,1H) ,2.98–2.80(m,4H),2.72(s,6H),2.66–2.59(m,2H),2.46–2.39(m,2H),2. 09–1.94(m,4H),1.89–1.77(m,6H),1.45–1.30(m,2H),0.95–0.86(m,6H).

[0321] Compound 28-2: LC-Ms m / z (ESI): 688.4 [M+H] +

[0322] 1 H NMR (400MHz, CD3OD) δ7.81–7.70(m,2H),7.10–7.05(m,1H),6.96–6.57(m,2H),5.78–5.72(m,1H),5.67–5.58(m,1H),3.83(s,3H),3.26–3.19( m,1H),3.00–2.64(m,11H),2.50–2.42(m,3H),2.12–2.02(m,2H),2.00 –1.92(m,1H),1.91–1.74(m,7H),1.45–1.30(m,2H),0.93–0.85(m,6H).

[0323] Example 29:

[0324] Referring to the synthetic route and preparation method of compound 2, a crude product of compound 29 was obtained, and the crude product was separated and purified by prep-HPLC (instrument: Waters 2767 preparative chromatography column: SunFire@Prep C18 (19 mm × 150 mm); (mobile phase composition: mobile phase A: acetonitrile, mobile phase B: water (containing 0.1% TFA) to obtain the trifluoroacetate salt of compound 29-1 (36 mg, yield 10.7%, HPLC retention time: 4.011 min) and the trifluoroacetate salt of compound 29-2 (40 mg, yield 11.8%, HPLC retention time: 4.181 min).

[0325] Trifluoroacetate salt of compound 29-1:

[0326] 1 H NMR (400MHz, CD3OD) δ7.56(s,1H),7.35(d,1H),6.67(s,2H),5.85–5.77(m,1H),5.74–5.64(m,1H),5.55–5.35(m,1H),3.95–3.66(m,5H),3.55 –3.37(m,3H),3.15–3.04(m,1H),2.98–2.80(m,5H),2.58–2.30(m,4H) ,2.14–1.86(m,8H),1.84(s,3H),1.45–1.30(m,1H),1.00–0.90(m,6H).

[0327] LC-Ms m / z(ESI):700.3[M+H] +

[0328] Trifluoroacetate salt of compound 29-2:

[0329] 1 H NMR (400MHz, CD3OD) δ7.57(s,1H),7.43–7.34(m,1H),6.70(s,2H),5.85–5.77(m,1H),5.72–5.64(m,1H),5.56–5.35(m,1H),3.91–3.66(m,5H),3 .56–3.43(m,3H),3.10–2.83(m,6H),2.54–2.32(m,4H),2.14–2.01(m,2H ),1.95(s,6H),1.84–1.70(m,2H),1.37–1.21(m,1H),0.93–0.83(m,6H).

[0330] LC-Ms m / z(ESI):700.3[M+H]

[0331] Example 30:

[0332] Referring to the synthetic route and preparation method of compound 2, a crude product of compound 30 was obtained, and the crude product was separated and purified by prep-HPLC (instrument: Waters 2767 preparative chromatography column: SunFire@Prep C18 (19 mm × 150 mm); (mobile phase composition: mobile phase A: acetonitrile, mobile phase B: water (containing 0.1% TFA) to obtain the trifluoroacetate salt of compound 30-1 (17 mg, yield 7.3%, HPLC retention time: 4.066 min) and the trifluoroacetate salt of compound 30-2 (23 mg, yield 9.9%, HPLC retention time: 4.246 min).

[0333] Trifluoroacetate salt of compound 30-1: LC-Ms m / z (ESI): 688.3 [M+H] +

[0334] Trifluoroacetate salt of compound 30-2:

[0335] 1 H NMR (400MHz, CD3OD) δ7.57(s,1H),7.43–7.34(m,1H),6.88(d,2H),5.84–5.77(m,1H),5.72–5.63(m,1H),5.56–5.36(m,1H),3.94–3.58(m,2H),3 .55–3.43(m,3H),3.10–2.80(m,6H),2.57–2.33(m,4H),2.16–2.03(m,2H ),1.98(s,6H),1.86–1.69(m,2H),1.36–1.24(m,1H),0.94–0.83(m,6H).

[0336] LC-Ms m / z(ESI):688.3[M+H] +

[0337] Example 31:

[0338] Referring to the synthetic route and preparation method of compound 2, a crude product of compound 31 was obtained, and the crude product was separated and purified by prep-HPLC (instrument: Waters 2767 preparative chromatography column: SunFire@Prep C18 (19 mm × 150 mm); (mobile phase composition: mobile phase A: acetonitrile, mobile phase B: water (containing 0.1% ammonium acetate) to obtain compound 31-1 (55 mg, yield 19%, HPLC retention time: 4.509 min) and compound 31-2 (42 mg, yield 14.5%, HPLC retention time: 4.589 min).

[0339] Compound 31-1:

[0340] 1 H NMR (400MHz, CD3OD) δ7.85(s,1H),7.13–7.04(m,1H),6.99–6.91(m,1H),6.79(s,1H),5.75–5.64(m,2H),5.33–5.14(m,1H),3.28–3.15(m, 1H),3.15–2.91(m,6H),2.90–2.72(m,5H),2.48–2.40(m,2H),2.35–2 .00(m,4H),1.99–1.75(m,8H),1.48–1.35(m,1H),0.99–0.90(m,6H).

[0341] LC-Ms m / z(ESI):738.3[M+H] +

[0342] Compound 31-2:

[0343] 1 H NMR(400MHz,CD3OD)δ7.87(s,1H),7.16–7.08(m,1H),7.02–6.95(m,1H),6.87( s,1H),5.78–5.71(m,1H),5.70–5.62(m,1H),5.35–5.16(m,1H),3.28–3.18(m, 3H),3.18–2.95(m,5H),2.95–2.82(m,3H),2.72–2.63(m,1H),2.52–2.44(m,2H ),2.38–2.04(m,4H),1.97–1.71(m,8H),1.40–1.27(m,1H),0.92–0.83(m,6H).

[0344] LC-Ms m / z(ESI):738.3[M+H] +

[0345] Example 32:

[0346] Referring to the synthetic route and preparation method of compound 2, a crude product of compound 32 was obtained, and the crude product was separated and purified by prep-HPLC (instrument: Waters 2767 preparative chromatography column: SunFire@Prep C18 (19 mm × 150 mm); (mobile phase composition: mobile phase A: acetonitrile, mobile phase B: water (containing 0.1% ammonium acetate) to obtain compound 32-1 (15 mg, yield 7.7%, HPLC retention time: 4.483 min) and compound 32-2 (20 mg, yield 10.3%, HPLC retention time: 4.565 min).

[0347] Compound 32-1:

[0348] 1 H NMR(400MHz,CD3OD)δ7.85(s,1H),7.08–7.01(m,1H),6.87–6.77(m,2H),5 .74–5.63(m,2H),5.33–5.12(m,1H),3.81(s,3H),3.25–3.13(m,1H),3.12 –2.89(m,6H),2.89–2.72(m,5H),2.46–2.37(m,2H),2.34–1.98(m,4H),1. 97–1.89(m,2H),1.87–1.67(m,6H),1.47–1.36(m,1H),0.98–0.88(m,6H).

[0349] LC-Ms m / z(ESI):750.3[M+H] +

[0350] Compound 32-2:

[0351] 1H NMR(400MHz,CD3OD)δ7.86(s,1H),7.10–7.04(m,1H),6.89–6.83(m,2H),5.79–5.71 (m,1H),5.70–5.62(m,1H),5.36–5.15(m,1H),3.83(s,3H),3.30–3.17(m,3H),3.17 –2.94(m,5H),2.93–2.82(m,3H),2.70–2.61(m,1H),2.50–2.42(m,2H),2.39–2.01( m,4H),1.92–1.81(m,7H),1.81–1.71(m,1H),1.41–1.26(m,1H),0.93–0.83(m,6H).

[0352] LC-Ms m / z(ESI):750.3[M+H] +

[0353] Reference compound A: prepared according to WO2021076890A1.

[0354] Preparation of control compound B:

[0355] Step 1: Synthesis of B-1

[0356] Under nitrogen, 44 g (230 mg, 0.51 mmol), 2,6-dimethyl-4-fluorophenylboronic acid (128 mg, 0.77 mmol), and potassium phosphate (325 mg, 1.53 mmol) were added to 1,4-dioxane (2.5 mL) and water (0.25 mL), followed by Xphos-G2-Pd (40 mg, 0.051 mmol). The mixture was sealed and reacted at 100°C for 24 h. After cooling to room temperature, ethyl acetate (40 mL) was added, and the mixture was washed sequentially with water (20 mL) and saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and flash chromatography on a silica gel column afforded B-1 (210 mg, 83.77% yield).

[0357] LC-Ms m / z(ESI):392.6[M+H-Boc] + .

[0358] Step 2: Synthesis of B-2

[0359] Dissolve B-1 (210 mg, 0.43 mmol) in tetrahydrofuran (3 mL), add 4N hydrochloric acid in dioxane (2.5 mL), stir at room temperature for 3 h, and concentrate under reduced pressure to obtain the hydrochloride salt of crude B-2.

[0360] LC-Ms m / z(ESI):392.6[M+H] + .

[0361] Step 3: Synthesis of B-3

[0362] Under nitrogen, the crude hydrochloride of B-2, B-2' (143 mg, 0.41 mmol) was dissolved in dry DMF (3.0 mL), and HOBT (110 mg, 0.82 mmol), EDCI (160 mg, 0.82 mmol), and DIPEA (158 mg, 1.23 mmol) were added. The mixture was allowed to react at room temperature overnight, and ethyl acetate (60 mL) was added. The mixture was washed sequentially with water (20 mL × 2) and saturated brine (20 mL × 1), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography to obtain B-3 (270 mg).

[0363] LC-Ms m / z(ESI):722.3[M+H] + .

[0364] Step 4: Synthesis of compound B-4

[0365] B-3 (270 mg) was dissolved in 4 mL of tetrahydrofuran and 1 mL of water, and lithium hydroxide monohydrate (44 mg, 1.05 mmol) was added. The reaction was allowed to react at room temperature for 5 h. The pH was adjusted to 5-6 with 1N hydrochloric acid, and the mixture was concentrated under reduced pressure to obtain crude compound B-4. The crude product was separated and purified by prep-HPLC (instrument: Waters 2767 preparative column: SunFire@Prep C18 (19 mm × 150 mm); (mobile phase composition: mobile phase A: acetonitrile, mobile phase B: water (containing 0.1% ammonium acetate)) to obtain compound B-4-P1 (23 mg, two-step yield 12.36%) and compound B-4-P2 (30 mg, two-step yield 13.18%).

[0366] Among them, compound B-4-P2 is the control compound B.

[0367] Biological testing

[0368] 1. Integrin α4β7 Cell Adhesion Assay

[0369] MAdCAM-1 was prepared with TBS buffer to a final concentration of 2 μg / ml, and 50 μl of the solution was transferred to a 96-well plate and coated overnight at 4°C. The plate was washed three times with TBS buffer, and then 150 μl of blocking solution (TBS buffer containing 1% BSA) was added and blocked at 37°C for 1 hour. RPMI8866 cells were harvested, washed twice with DPBS buffer, and then resuspended in TBS buffer to 4 × 10 5cells / ml, transfer 50 μl of cell solution to a 96-well plate to make the cell density 2×10 5 cells / well, then add 1 μl of different concentrations of compound or DMSO, and incubate at 37°C for 1 hour or 2 hours. Wash the plate with TBS buffer to remove non-adherent cells, add 50 μl of substrate (4-nitrophenyl-N-acetyl-β-D-glucosaminide), incubate at 37°C for 2 hours, and finally add 90 μl of stop solution (50 mM glycine and 5 mM EDTA, pH 10.4). Read the OD value of the plate at 405 nm using a microplate reader. Calculate the IC using GraphPad Prism 6 software. 50 value.

[0370] 2. Integrin α4β1 Cell Adhesion Assay

[0371] VCAM was prepared with coating buffer to a final concentration of 0.5 μg / ml. 50 μl of the solution was transferred to a 96-well plate and coated overnight at 4°C. The plate was washed three times with coating buffer, then 150 μl of blocking buffer (containing 1% BSA) was added and blocked at 37°C for 1 hour. Jurkat cells were harvested, washed twice with DPBS, and resuspended in the reaction buffer to 4 × 106 cells / ml. 50 μl of the cell solution was transferred to a 96-well plate at a cell density of 2 × 105 cells / well. 1 μl of test compound or DMSO at various concentrations was added and incubated at 37°C for 1 hour. Wash the plate with coating buffer to remove non-adherent cells, add 50 μl of substrate (4-nitrophenyl-N-acetyl-β-D-glucosaminide), incubate at 37°C for 2 h, and finally add 90 μl of stop solution (50 mM glycine and 5 mM EDTA, pH 10.4) and read the OD value of the plate at 405 nm.

[0372] Coating buffer: DMEM + 20mM HEPES + 0.4mM MnCl2

[0373] Buffer: DMEM + 20mM HEPES + 0.1% BSA + 0.4mM MnCl2

[0374] DMEM:dulbecco's modified eagle medium;

[0375] HEPES:2-[4-(2-hydroxyethyl)piperazin-1-yl]ethanesulfonic acid

[0376] Table 1 Test results of compound adhesion to α4β7 and α4β1 cells

[0377] Conclusion: The compounds of the present invention, such as the compounds in the examples, have a good inhibitory effect on cell adhesion mediated by integrin α4β7 and have good selectivity for α4β1 / α4β7.

[0378] 3. CYP450 enzyme inhibition test

[0379] The purpose of this study was to evaluate the effects of test substances on the activities of five cytochrome P450 (CYP) isoforms (CYP1A2, CYP2C9, CYP2D6, and CYP3A4) in human liver microsomes using an in vitro assay system. Specific probe substrates for each CYP450 isoform were incubated with human liver microsomes and varying concentrations of the test substances. The reactions were initiated by the addition of reduced nicotinamide adenine dinucleotide phosphate (NADPH). Following the reaction, samples were processed and metabolites generated from the specific substrates were quantitatively detected by liquid chromatography-tandem mass spectrometry (LC-MS / MS). Changes in CYP enzyme activity were measured, and IC50 values ​​were calculated to evaluate the inhibitory potential of the test substances against each CYP isoform, CYP1A2, CYP2C9, CYP2D6, and CYP3A4-M (using midazolam as a substrate).

[0380] Conclusion: The compounds of the present invention, such as the compounds in the examples, have no significant inhibitory effect on any subtype of CYP enzymes.

[0381] 4. Pharmacokinetic Testing in Mice

[0382] Experimental animals: Male BALB / c mice, 20-25 g, 6 mice per compound, purchased from Chengdu Dashuo Experimental Animal Co., Ltd.

[0383] Experimental Design: On the day of the experiment, mice were randomly divided into groups according to body weight. They were fasted but not watered for 12-14 hours before administration and fed 4 hours after administration.

[0384] Table 2 Dosage information

[0385] Note: Intravenous administration solvent: 5% DMA + 5% Solutol + 90% Saline; Oral administration solvent: 5% DMSO + 95% (20% SBE-β-CD in saline)

[0386] (DMA: dimethylacetamide; Solutol: polyethylene glycol-15-hydroxystearate; Saline: normal saline;

[0387] DMSO: dimethyl sulfoxide; SBE-β-CD: sulfobutyl-β-cyclodextrin)

[0388] Before and after drug administration, 0.06 mL of blood was collected via the orbital cavity under isoflurane anesthesia. The blood was placed in an EDTAK2 centrifuge tube and centrifuged at 5000 rpm at 4°C for 10 minutes to collect plasma. Blood was collected from both the intravenous and oral gavage groups at 0, 5, 15, 30 minutes, and 1, 2, 4, 6, 8, and 24 hours. All samples were stored at -80°C prior to analysis and quantitative analysis was performed using LC-MS / MS.

[0389] Table 3 Mouse PK results

[0390] Conclusion: The compounds of the present invention, such as the compounds in the examples, have good oral absorption properties in mice.

[0391] 5. Pharmacokinetic Test in Rat

[0392] Experimental animals: Male SD rats, about 220 g, 6 to 8 weeks old, 6 rats per compound, purchased from Chengdu Dashuo Experimental Animal Co., Ltd.

[0393] Experimental Design: On the day of the experiment, 6 SD rats / compound were randomly divided into groups according to body weight. They were fasted but not watered for 12-14 hours before administration and fed 4 hours after administration.

[0394] Table 4 Dosing information

[0395] Note: Intravenous administration solvent: 5% DMA + 5% Solutol + 90% Saline; Oral administration solvent: 5% DMSO + 95% (20% SBE-β-CD in saline)

[0396] (DMA: dimethylacetamide; Solutol: polyethylene glycol-15-hydroxystearate; Saline: normal saline;

[0397] DMSO: dimethyl sulfoxide; SBE-β-CD: sulfobutyl-β-cyclodextrin)

[0398] Before and after drug administration, 0.10 mL of blood was collected via the orbital cavity under isoflurane anesthesia. The blood was placed in an EDTAK2 centrifuge tube and centrifuged at 5000 rpm at 4°C for 10 minutes to collect plasma. Blood was collected from both the intravenous and oral gavage groups at 0, 5, 15, 30 minutes, and 1, 2, 4, 6, 8, and 24 hours. All samples were stored at -80°C prior to analysis and quantitative analysis was performed using LC-MS / MS.

[0399] Table 5 Rat PK results

[0400] Conclusion: The compounds of the present invention, such as the compounds in the examples, have better pharmacokinetic properties in rats (such as lower clearance, better Cmax, AUC and bioavailability).

[0401] 6. Pharmacokinetic Testing in Beagle Dogs

[0402] Experimental animals: Male beagle dogs, weighing about 8-11 kg, 5-6 per compound, purchased from Beijing Masi Biotechnology Co., Ltd.

[0403] Test method: On the test day, 5-6 beagle dogs were randomly divided into groups according to body weight. The dogs were fasted but not watered for 12-14 hours before administration and fed 4 hours after administration.

[0404] Table 6 Dosage information

[0405] Note: Intravenous administration solvent: 5% DMA + 5% Solutol + 90% Saline; Oral administration solvent: 5% DMSO + 95% (20% SBE-β-CD in saline)

[0406] (DMA: dimethylacetamide; Solutol: polyethylene glycol-15-hydroxystearate; Saline: normal saline;

[0407] DMSO: dimethyl sulfoxide; SBE-β-CD: sulfobutyl-β-cyclodextrin)

[0408] Before and after administration, 1 ml of blood was collected from the jugular vein or limb vein and placed in an EDTAK2 centrifuge tube.

[0409] Plasma was collected by centrifugation at 5000 rpm and 4°C for 10 minutes. Blood was collected from both the intravenous and oral gavage groups in groups G1 and G2 at the following time points: 0, 5, 15, 30 minutes, 1, 2, 4, 6, 8, 10, 12, 24 hours, 48, and 72 hours. Blood was collected from both the intravenous and oral gavage groups in groups G3 and G4 at the following time points: 0, 5, 15, 30 minutes, 1, 2, 4, 6, 8, 10, 12, and 24 hours. All samples were stored at -80°C prior to analysis and quantitative analysis was performed using LC-MS / MS.

[0410] Conclusion: The compounds of the present invention, such as the compounds in the examples, have good oral absorption properties in beagle dogs.

[0411] 7. Mouse Liver Microsome Stability Test

[0412] At 37°C, 1 μM of the test compound was incubated with mouse liver microsomes (0.5 mg / mL) supplemented with an NADPH regeneration system for 5, 10, 20, 30, and 60 minutes. The concentration of the test compound in the resulting samples was determined by LC-MS / MS. The half-life (T) of the compound in the mouse liver microsome solution was obtained by calculating the remaining percentage of the compound at each time point. 1 / 2 ) and intrinsic clearance (CL int(mic) ).

[0413] Biological test results:

[0414] Table 7 Mouse liver microsome stability test results

[0415] Conclusion: The compounds of the present invention, such as the compounds in the examples, have good stability in mouse liver microsomes.

[0416] 8.Caco2 permeability test

[0417] The experiment used Caco-2 cell monolayers in 96-well Transwell plates, incubated in triplicate. Transport buffer (HBSS, 10 mM HEPES, pH 7.4 ± 0.05) containing the compound of the invention (2 μM) or the control compounds digoxin (10 μM), nadolol (2 μM), and metoprolol (2 μM) was added to the apical or basolateral wells of the cell monolayer. Transport buffer containing DMSO was added to the corresponding receiving wells. After incubation at 37 ± 1°C for 2 hours, the cell plate was removed and appropriate samples were taken from the apical and basolateral wells to a new 96-well plate. Proteins were then precipitated by adding acetonitrile containing an internal standard. Samples were analyzed using LC-MS / MS to determine the concentrations of the compound of the invention and the control compound. The concentration data were used to calculate the apparent permeability coefficients for transport from the apical to basolateral side of the cell monolayer and from the basolateral to apical side, thereby calculating the efflux rate. The integrity of the cell monolayer after 2 hours of incubation was assessed by leakage of Lucifer Yellow.

[0418] Table 8 Caco2 test results

[0419] Conclusion: The compounds of the present invention, such as the compounds in the examples, have good permeability.

[0420] 9. CCK2 agonist activity test

[0421] CHO cells overexpressing human CCK2 were cultured in F-12 medium (supplemented with 10% fetal bovine serum and 0.2 mg / mL Hygromycin B) at 37°C in 5% CO2 until use. 1× Stimulation Buffer was prepared according to the instructions for the HTRF IP-One Gq Detection Kit (Revvity, Cat. No. 62IPAPEJ). A 1000× working solution of the test compound was prepared in DMSO and transferred using an ECHO655 to a 384-well plate (Greiner, Cat. No. 784075). CHO cells overexpressing human CCK2 were trypsinized and centrifuged at 1000 rpm for 5 minutes. The cell pellet was resuspended in 1× Stimulation Buffer and, after counting, 10 μL of the solution was added to a 384-well plate (Greiner, Cat. No. 784075) at a density of 8,000 cells per well. Centrifuge at 1000 rpm for 1 minute at room temperature and incubate at 37°C for 60 minutes. Dilute IP1-d2 to 7× working concentration with Lysis & Detection Buffer, take 2 μL and add to the corresponding experimental wells. Then dilute Anti-IP1-Cryptate to 7× working concentration with Lysis & Detection Buffer, take 2 μL and add to the corresponding experimental wells. Centrifuge at 1000 rpm for 1 minute and incubate at room temperature in the dark for 1 hour. After incubation, use The FSX microplate reader (BMG) was used to detect the fluorescence at 665 nm and 620 nm under the excitation wavelength of 330 nm. The signal ratio (Signal 665 / Signal 620*10 4 ), and EC was fitted using Graphpad Prism software. 50 value.

[0422] Data Analysis:

[0423] Data were analyzed and processed using GraphPad Prism and Excel software. The degree of activation of M1-M5 targets by different compound concentrations was calculated using the following formula:

[0424] Activation% represents the percentage of the compound's agonism on the receptor. Mean values ​​of positive controls. Mean values ​​of negative controls (DMSO).

[0425] Compound EC 50 Calculated using GraphPad Prism software by fitting the following equation:

[0426] Y=Bottom+(Top-Bottom) / (1+10^((LogEC 50 -X)*HillSlope))

[0427] Where X is the test concentration of the test article, Y is the activation percentage at the corresponding concentration, and Bottom and Top are the minimum and maximum activation percentages, respectively.

[0428] Conclusion: The compounds of the present invention, such as the compounds in the examples, have weak agonist activity on CCK2.

[0429] The above describes exemplary embodiments of the present invention. However, the scope of protection of the present invention is not limited to the above embodiments. Any modifications, equivalent substitutions, improvements, etc. made by those skilled in the art within the spirit and principles of the present invention should be included in the scope of protection of the present invention.

Claims

1. A compound or its stereoisomer, racemate, deuterated form, solvate, prodrug, metabolite, pharmaceutically acceptable salt or cocrystal, the compound being selected from the compound represented by general formula (I), wherein R 1 Selected from H or C 1-4 alkyl; R 2 Selected from halogen, OH, cyano, C 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 1-6 Alkoxy, C 3-6 Cycloalkyl or 4 to 7 membered heterocyclic group, wherein the alkyl, alkenyl, alkynyl, alkoxy, cycloalkyl or heterocyclic group is optionally substituted by 1 to 4 atoms selected from deuterium, halogen, CN, C 1-6 Alkyl or C 1-6 substituted by an alkoxy substituent; R 2a Each independently selected from halogen, OH, cyano, C 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 1-6 Alkoxy, C 3-6 Cycloalkyl or 4 to 7 membered heterocyclic group, wherein the alkyl, alkenyl, alkynyl, alkoxy, cycloalkyl or heterocyclic group is optionally substituted by 1 to 4 atoms selected from deuterium, halogen, CN, C 1-6 Alkyl or C 1-6 substituted by an alkoxy substituent; R 3 Selected from R 3a 、R 3b are independently selected from H, halogen, cyano, C 1-6 Alkyl, C 1-6 Alkoxy, the alkyl or alkoxy is optionally substituted by 1 to 4 atoms selected from deuterium, halogen, CN, C 1-6 Alkyl or C 1-6 substituted by an alkoxy substituent; n is selected from 0, 1, 2, 3 or 4; Provided that the compound represented by general formula (I) is not the following compound and its stereoisomers 2. The compound according to claim 1 or its stereoisomer, racemate, deuterated form, solvate, prodrug, metabolite, pharmaceutically acceptable salt or cocrystal, wherein: Selected from R 1 Selected from H, methyl or ethyl; R 2 Selected from F, methyl, CHF2, CH2F, CF3; R 2a Each is independently selected from F, Cl, Br, OH, cyano, methyl, ethyl, isopropyl, methoxy, ethoxy or cyclopropyl, wherein the methyl, ethyl, isopropyl, methoxy, ethoxy or cyclopropyl is optionally substituted by 1 to 4 groups selected from deuterium, halogen, CN, C 1-4 Alkyl or C 1-4 substituted by an alkoxy substituent; R 3a 、R 3b are each independently selected from H, F, Cl, Br, cyano, methyl, ethyl, isopropyl, methoxy or ethoxy, wherein the methyl, ethyl, isopropyl, methoxy or ethoxy is optionally substituted by 1 to 4 atoms selected from deuterium, halogen, CN, C 1-4 Alkyl or C 1-4 substituted by an alkoxy substituent.

3. The compound according to claim 2, or a stereoisomer, racemate, deuterated form, solvate, prodrug, metabolite, pharmaceutically acceptable salt or cocrystal thereof, R 3 Selected from Selected from 4. The compound according to claim 1 or its stereoisomer, racemate, deuterated form, solvate, prodrug, metabolite, pharmaceutically acceptable salt or cocrystal, wherein the compound is selected from one of the structures shown in Table E-1.

5. A pharmaceutical composition comprising a compound according to any one of claims 1 to 4 or a stereoisomer, racemate, deuterated form, solvate, prodrug, metabolite, pharmaceutically acceptable salt or cocrystal thereof, and a pharmaceutically acceptable carrier. Preferably, the pharmaceutical composition comprises 1 to 1500 mg of the compound according to any one of claims 1 to 4 or a stereoisomer, racemate, deuterated form, solvate, prodrug, metabolite, pharmaceutically acceptable salt or cocrystal thereof.

6. Use of the compound according to any one of claims 1 to 4 or its stereoisomer, racemate, deuterated form, solvate, prodrug, metabolite, pharmaceutically acceptable salt or cocrystal, or the pharmaceutical composition according to claim 5 in the preparation of a medicament for treating diseases associated with α4β7 activity or expression.

7. The use according to claim 6, characterized in that The disease is selected from intestinal inflammatory diseases (preferably Crohn's disease or ulcerative colitis).

8. A method for treating a disease in a mammal, the method comprising administering to a subject a therapeutically effective amount of the compound of any one of claims 1 to 4, or a stereoisomer, racemate, deuterated form, solvate, prodrug, metabolite, pharmaceutically acceptable salt or cocrystal thereof, or the pharmaceutical composition of claim 5, wherein the therapeutically effective amount is preferably 1-1500 mg, and the disease is preferably an intestinal inflammatory disease (preferably Crohn's disease or ulcerative colitis).

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