Pharmaceutical composition of five-membered and five-membered ring derivative and application of pharmaceutical composition in medicine

Abstract

The invention relates to a pharmaceutical composition or a pharmaceutical preparation, which comprises an active ingredient A with a therapeutically effective amount and a pharmaceutical excipient, the active ingredient A is selected from a compound as shown in a general formula (I) or a stereoisomer, a tautomer, a deuterated compound, a solvate, a prodrug, a metabolite, a pharmaceutically acceptable salt or a co-crystal of the compound, and the pharmaceutical composition or the pharmaceutical preparation contains 1-600mg of the active ingredient A. The invention also relates to application of the pharmaceutical composition or the pharmaceutical preparation in preparation of related drugs for treating diabetes. # imgabs0 #

Description

A pharmaceutical composition of a five-membered five-ring derivative and its application in medicine Technical Field

[0001] The present invention belongs to the field of pharmaceutical preparations and specifically relates to a pharmaceutical composition or pharmaceutical preparation comprising a therapeutically effective amount of an active ingredient A and a pharmaceutical excipient. The active ingredient A is selected from a compound of general formula (I) or a stereoisomer, tautomer, deuterated form, solvate, prodrug, metabolite, pharmaceutically acceptable salt, or cocrystal thereof. The pharmaceutical composition or pharmaceutical preparation comprises 1-600 mg of the active ingredient A. The present invention also relates to the use of the pharmaceutical composition or pharmaceutical preparation in preparing a drug for treating diabetes. Background Art

[0002] Diabetes is a group of metabolic diseases characterized by hyperglycemia. Hyperglycemia is caused by defects in insulin secretion, impaired insulin action, or both. Long-term high blood sugar levels in diabetes lead to chronic damage and dysfunction of various tissues, particularly the eyes, kidneys, heart, blood vessels, and nerves. Diabetes is primarily divided into two types. Type 1 diabetes: Destruction of pancreatic B cells leads to absolute insulin deficiency. Type 2 diabetes: Insulin resistance with relative insulin deficiency or impaired insulin secretion with insulin resistance.

[0003] Drugs for type 2 diabetes can be divided into six major categories (insulin, insulin secretagogues, biguanides, glucosidase inhibitors, thiazolidinediones, and SGLT2 inhibitors), each of which works through a different primary mechanism. However, with the exception of GLP-1 receptor agonists and SGLT2 inhibitors, these drugs have limited efficacy and do not address the most important issue, namely, decreased cellular function and associated obesity.

[0004] GLP-1 is a 30-amino acid long incretin hormone secreted by L cells in the intestine. GLP-1 stimulates insulin secretion in a physiological, glucose-dependent manner, reduces glucagon secretion, inhibits gastric emptying, reduces appetite, and stimulates β-cell proliferation. In nonclinical studies, GLP-1 has been shown to promote β-cell sustainability by stimulating the transcription of genes important for glucose-dependent insulin secretion and promoting β-cell neogenesis. In healthy individuals, GLP-1 plays a key role in regulating postprandial blood flow, increasing peripheral glucose absorption by stimulating glucose-dependent insulin secretion from the pancreas. GLP-1 also inhibits glucagon secretion, resulting in reduced hepatic glucose output. Furthermore, GLP-1 delays gastric emptying, slows small intestinal motility, and delays food absorption.

[0005] GLP-1 receptor agonists, such as GLP-1, liraglutide, and exendin-4, are peptide drugs, mostly administered by injection. Small molecule GLP-1 receptor agonists have become a hot topic in drug development in recent years due to their potential for high oral bioavailability.

[0006] Summary of the Invention

[0007] The present invention provides a pharmaceutical composition or pharmaceutical preparation comprising a therapeutically effective amount of an active ingredient A and a pharmaceutical excipient, wherein the active ingredient A is selected from a compound of formula (I) or a stereoisomer, tautomer, deuterated form, solvate, prodrug, metabolite, pharmaceutically acceptable salt, or cocrystal thereof, and wherein the pharmaceutical composition or pharmaceutical preparation has a dosage of 1-600 mg. The present invention also relates to the use of the pharmaceutical composition or pharmaceutical preparation in the preparation of a medicament for treating diabetes.

[0008] The compound of the present invention has good oral performance, good safety and hypoglycemic activity.

[0009] The present invention relates to a pharmaceutical composition or pharmaceutical preparation, comprising a therapeutically effective amount of active ingredient A and a pharmaceutical excipient. The pharmaceutical composition can be in the form of a unit preparation.

[0010] The present invention relates to a pharmaceutical composition or pharmaceutical preparation, wherein the pharmaceutical composition or pharmaceutical preparation comprises an active ingredient A and a pharmaceutical excipient, wherein the active ingredient A is selected from the compound of general formula (I) or its stereoisomers, tautomers, deuterated substances, solvates, prodrugs, metabolites, pharmaceutically acceptable salts or cocrystals.

[0011] In some embodiments, Ring B is selected from C 6-10 Aromatic ring or 5 to 12 membered heteroaromatic ring, said aromatic ring or heteroaromatic ring is optionally substituted by 1 to 4 R B Substitution, the heterocyclic group or heteroaromatic ring contains 1 to 3 heteroatoms selected from O, S and N;

[0012] In some embodiments, Ring B is selected from one of the following groups, which may be substituted or unsubstituted: Its left side is directly connected to ring C and, when substituted, is optionally substituted by 1, 2 or 3 R B replace;

[0013] In some embodiments, ring C is selected from C 6-10 Carbocyclic, 5- to 10-membered heterocyclic, C 6-10aromatic ring or 5 to 10 membered heteroaromatic ring, wherein the carbocyclic ring, heterocyclic ring, aromatic ring or heteroaromatic ring is optionally substituted by 1 to 4 R C Substituted, the heterocyclic or heteroaromatic ring contains 1 to 5 heteroatoms selected from O, S and N;

[0014] In some embodiments, Ring C is selected from one of the following groups, which are substituted or unsubstituted: When substituted, it is optionally replaced by 1, 2 or 3 R C Replace, its left side is connected to L;

[0015] In some embodiments, L is selected from a bond, O, S, -NR L -、-C(R L )2-、-C(R L )2-C(R L )2-、-YC(R L )2-、-C(R L )2-Y-、-YC(R L )2-C(R L )2-、-C(R L )2-C(R L )2-Y-or-C(R L )2-YC(R L )2-;

[0016] In some embodiments, L is selected from -CH2O-, -OCH2-, -CH2NH-, -NHCH2-, -CH2CH2-, -OCH2CH2-, -CH2CH2O-, -NHCH2CH2-, -CH2CH2NH-, and the right side is connected to ring C;

[0017] In some embodiments, or Selected from Its right side is connected to ring B, wherein the benzene ring is optionally substituted by 1, 2 or 3 R C replace;

[0018] In some embodiments, Y is selected from O, S, or -NR L -;

[0019] In some embodiments, R1 is selected from H, halogen, OH, -SH, CF3, CN, C 1-6 Alkyl, C 1-6 Alkoxy, -C 1-3 Alkylene-ZC 0-3 Alkylene-R 1a 、C 0-4 Alkylene-R 1a The alkyl, alkoxy, and alkylene groups are optionally substituted by 1 to 4 groups selected from halogen, =O, CN, OH, -N(R1b )2、C 1-6 Alkyl, halogen-substituted C 1-6 Alkyl, hydroxy substituted C 1-6 Alkyl, cyano substituted C 1-6 Alkyl, C 1-6 Alkoxy, C 3-6 substituted by a cycloalkyl, a 3- to 6-membered heterocycloalkyl, or a 5- to 10-membered heteroaryl substituent, wherein the heterocycloalkyl or heteroaryl group contains 1 to 3 heteroatoms selected from O, S, and N;

[0020] In some embodiments, R1 is selected from

[0021] In some embodiments, Z is selected from a bond, N(R 1b ), O or S;

[0022] In some embodiments, R 1a Selected from C 1-6 Alkyl, C 1-6 Alkoxy, C 3-8 Cycloalkyl, 3 to 8 membered heterocycloalkyl, 6 to 10 membered aryl or 5 to 12 membered heteroaryl, wherein the cycloalkyl, heterocycloalkyl, aryl, heteroaryl is optionally substituted by 1 to 4 groups selected from halogen, =O, OH, CN, C 1-6 Alkyl, C 1-6 Alkoxy or -N(R 1b ) 2 is substituted by a substituent, wherein the alkyl group or alkoxy group is optionally substituted by 1 to 4 substituents selected from halogen, =O, CN, OH, -N(R 1b )2、C 1-6 Alkyl, C 1-6 Alkoxy, C 3-6 Substituted by a cycloalkyl substituent, wherein the heterocycloalkyl or heteroaryl group contains 1 to 3 heteroatoms selected from O, S, and N;

[0023] In some embodiments, R 1b Each independently selected from H or C 1-6 Alkyl, said alkyl being optionally substituted by 1 to 3 groups selected from halogen, =O, CN, OH, NH2, C 1-6 Alkyl, C 1-6 Alkoxy or C 3-6 substituted by a cycloalkyl substituent;

[0024] In some embodiments, R L Each independently selected from H, halogen, -SH, CN, OH, CF3, NH2, C 1-6 Alkyl, C 1-6 Alkoxy or C 3-6Cycloalkyl, wherein the alkyl, cycloalkyl or alkoxy is optionally substituted by 1 to 4 groups selected from halogen, =O, OH, CN, NH2, C 1-6 Alkyl, C 3-6 Cycloalkyl or C 1-6 substituted by an alkoxy substituent;

[0025] In some embodiments, R B 、R C or R D Each independently selected from H, halogen, =O, CN, CF3, OH, -SH, NH2, C 1-6 Alkyl, C 3-6 Cycloalkyl or C 1-6 Alkoxy, wherein the alkyl, cycloalkyl or alkoxy is optionally substituted by 1 to 4 groups selected from halogen, =O, OH, CN, NH2, C 1-6 Alkyl, C 3-6 Cycloalkyl or C 1-6 substituted by an alkoxy substituent;

[0026] In some embodiments, R B 、R C or R D Each independently selected from H, F, Cl, CN, CF3, CHF2, methyl, ethyl, methoxy, ethoxy, tert-butyloxy, trifluoromethoxy;

[0027] Alternatively, R C With R L Each of the atoms, both independent and connected, forms a C 3-10 Carbocyclic or 3 to 10 membered heterocyclic ring, said carbocyclic or heterocyclic ring being optionally substituted by 1 to 4 rings selected from halogen, =O, OH, CN, NH2, C 1-6 Alkyl, C 3-6 Cycloalkyl or C 1-6 Alkoxy is substituted by a substituent, wherein the alkyl, cycloalkyl, alkoxy is optionally substituted by 1 to 4 groups selected from halogen, =O, OH, CN, NH2, C 1-6 Alkyl, C 3-6 Cycloalkyl or C 1-6 substituted by an alkoxy substituent, wherein the heterocyclic ring contains 1 to 3 heteroatoms selected from O, S and N;

[0028] In some embodiments, m is selected from 0, 1, 2, 3, or 4;

[0029] In some embodiments, the structure of the compound is selected from one of the structures shown in Table S-1;

[0030] Table S-1 Compound Structure

[0031] In some embodiments, the pharmaceutically acceptable salt is selected from the group consisting of tris salts;

[0032] In some embodiments, the pharmaceutically acceptable salt is selected from a tris salt or a triethanolamine salt.

[0033] In some embodiments, active ingredient A is selected from tris(hydroxymethyl)aminomethane salts of the following structure,

[0034] In some embodiments, active ingredient A is selected from triethanolamine salts of the following structure,

[0035] The present invention relates to a pharmaceutical composition or pharmaceutical preparation comprising the active ingredient A in any one of the aforementioned embodiments and a pharmaceutically acceptable excipient.

[0036] In some embodiments, the pharmaceutically acceptable excipient comprises hydroxypropyl-β-cyclodextrin;

[0037] In some embodiments, the content of active ingredient A is 0.5%-15.4%;

[0038] In some embodiments, the pharmaceutical excipient comprises lactose (preferably lactose monohydrate) or sodium lauryl sulfate, and optionally, the content of active ingredient A is 20%-25%;

[0039] In some embodiments, the content of active ingredient A is 20%-25%;

[0040] In some embodiments, the pharmaceutical excipient comprises lactose (preferably lactose monohydrate) or sodium lauryl sulfate, and further contains one or more of microcrystalline cellulose, croscarmellose sodium, silicon dioxide, or magnesium stearate;

[0041] In some embodiments, the pharmaceutical excipient comprises one or more of microcrystalline cellulose, croscarmellose sodium, silicon dioxide, or magnesium stearate;

[0042] In some embodiments, the pharmaceutical composition or pharmaceutical formulation comprises 1-600 mg of active ingredient A;

[0043] In some embodiments, the pharmaceutical composition or pharmaceutical formulation comprises 5-300 mg of active ingredient A;

[0044] In some embodiments, the pharmaceutical composition or pharmaceutical formulation comprises 5-200 mg of active ingredient A;

[0045] In some embodiments, the pharmaceutical composition or pharmaceutical formulation comprises 5-100 mg of active ingredient A;

[0046] In some embodiments, the pharmaceutical composition or pharmaceutical formulation comprises 5 mg of active ingredient A;

[0047] In some embodiments, the pharmaceutical composition or pharmaceutical formulation comprises 10 mg of active ingredient A;

[0048] In some embodiments, the pharmaceutical composition or pharmaceutical formulation comprises 20 mg of active ingredient A;

[0049] In some embodiments, the pharmaceutical composition or pharmaceutical formulation comprises 25 mg of active ingredient A;

[0050] In some embodiments, the pharmaceutical composition or pharmaceutical formulation comprises 30 mg of active ingredient A;

[0051] In some embodiments, the pharmaceutical composition or pharmaceutical formulation comprises 40 mg of active ingredient A;

[0052] In some embodiments, the pharmaceutical composition or pharmaceutical formulation comprises 50 mg of active ingredient A;

[0053] In some embodiments, the pharmaceutical composition or pharmaceutical formulation comprises 75 mg of active ingredient A;

[0054] In some embodiments, the pharmaceutical composition or pharmaceutical formulation comprises 100 mg of active ingredient A;

[0055] In some embodiments, the pharmaceutical composition or pharmaceutical formulation comprises 125 mg of active ingredient A;

[0056] In some embodiments, the pharmaceutical composition or pharmaceutical formulation comprises 150 mg of active ingredient A.

[0057] In some embodiments, the pharmaceutical composition or pharmaceutical formulation comprises 200 mg of active ingredient A.

[0058] In some embodiments, the pharmaceutical composition or pharmaceutical formulation comprises 250 mg of active ingredient A.

[0059] In some embodiments, the pharmaceutical composition or pharmaceutical formulation comprises 350 mg of active ingredient A.

[0060] The present invention provides a tris(hydroxymethyl)aminomethane salt of the compound shown below and a solvate thereof:

[0061] In some embodiments, the molar ratio of compound: tris(hydroxymethyl)aminomethane is 1:0.9 to 1:1.1, preferably 1:1;

[0062] In some embodiments, the molar ratio of compound:tris(hydroxymethyl)aminomethane is 1:1;

[0063] In some embodiments, the solvate is methanol;

[0064] The present invention shows the crystalline form A of the tris(hydroxymethyl)aminomethane salt of the compound shown below:

[0065] Among them, using Cu-Kα radiation, its X-ray powder diffraction pattern has characteristic diffraction peaks at the following 2θ positions: 8.97°±0.2°, 9.32°±0.2°, 9.92°±0.2°, 10.68°±0.2°, 17.50°±0.2°, 20.17°±0.2°;

[0066] In some embodiments, the tris(hydroxymethyl)aminomethane salt crystalline form A of the aforementioned compound, using Cu-Kα radiation, has an X-ray powder diffraction pattern having characteristic diffraction peaks at the following 2θ positions: 8.97°±0.2°, 9.32°±0.2°, 9.92°±0.2°, 10.68°±0.2°, 15.07°±0.2°, 17.50°±0.2°, 17.71°±0.2°, 19.95°±0.2°, 20.17°±0.2°, 24.27°±0.2°, and 26.81°±0.2°;

[0067] In some embodiments, the tris(hydroxymethyl)aminomethane salt crystalline form A of the aforementioned compound, using Cu-Kα radiation, has an X-ray powder diffraction pattern having characteristic diffraction peaks at the following 2θ positions: 6.02°±0.2°, 8.97°±0.2°, 9.32°±0.2°, 9.92°±0.2°, 10.68°±0.2°, 12.03±0.2°, 13.77°±0.2°, 15.07°±0.2°, 16.18°±0.2°, 17.50°±0.2°, 17.71° ±0.2°, 18.27°±0.2°, 18.70°±0.2°, 19.15°±0.2°, 19.95°±0.2°, 20.17°±0.2°, 20.90°±0.2°, 21.89°±0.2°, 22.35°±0.2°, 24.05°±0.2°, 24.27°±0.2°, 24.65°±0.2°, 25.03°±0.2°, 25.54°±0.2°, 26.81°±0.2°, 28.03°±0.2°;

[0068] In some embodiments, the tris(hydroxymethyl)aminomethane salt crystalline form A of the aforementioned compound, using Cu-Kα radiation, has an X-ray powder diffraction pattern as shown in FIG1 ;

[0069] In some embodiments, the triethanolamine salt of the compound represented by the aforementioned formula (I) in Form A has a differential scanning calorimetry (DSC) curve showing an endothermic curve, wherein T 初始 =146.2℃, T 峰值 =149.9°C, ΔH=73.6 J / g, and its thermogravimetric analysis curve (TGA) showed a weight loss of 1% before 120°C.

[0070] In some embodiments, the differential scanning calorimetry curve and thermogravimetric analysis curve of the tris(hydroxymethyl)aminomethane salt crystalline form A of the compound represented by formula (I) are shown in FIG2 .

[0071] The present invention also provides a method for treating a disease in a mammal, comprising administering to the mammal a therapeutically effective amount of the compound or pharmaceutical composition disclosed herein, preferably 1-600 mg. The disease is preferably diabetes.

[0072] The present invention also provides use of the pharmaceutical composition or pharmaceutical preparation, the salt of the aforementioned compound and its solvate, and the crystal form A of the aforementioned compound in preparing drugs for treating diabetes.

[0073] As used herein, "effective amount" or "therapeutically effective amount" refers to a sufficient amount of the compound disclosed herein to alleviate to some extent one or more symptoms of the disease or disorder (eg, diabetes) being treated.

[0074] In some embodiments, the result is reduction and / or alleviation of the signs, symptoms, or causes of a disease, or any other desired alteration of a biological system.

[0075] For example, an "effective amount" for therapeutic uses is the amount of a composition comprising a compound disclosed herein that provides a clinically significant reduction in disease symptoms. Examples of therapeutically effective amounts include, but are not limited to, 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, 2-500 mg, 3-500 mg, 4-500 mg, 5-500 mg, 6-500 mg, 10-500 mg, mg, 20-500mg, 25-500mg, 30-500mg, 40-500mg, 50-500mg, 60-500mg, 70-500mg, 75-500mg, 80-500mg, 90-500mg, 100-500mg, 125-500mg, 150-5 00mg, 200-500mg, 250-500mg, 300-500mg, 400-500mg, 5-400mg, 10-400mg, 20-400mg, 25-400mg, 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, 125-300m g, 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, 1-100mg, 2-100mg, 5-100mg, 10-100mg, 15-100mg, 20-100mg,In some embodiments, examples of therapeutically effective amounts include, but are not limited to, 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, 125 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. 、

[0076] In some embodiments, the formulation specifications of the pharmaceutical composition or pharmaceutical preparation of the present invention include but are not limited to 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, 2-500 mg, 3-500 mg, 4-500 mg, 5-500 mg, 6-500 mg , 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, 150-5 00mg, 200-500mg, 250-500mg, 300-500mg, 400-500mg, 5-400mg, 10-400mg, 20-400mg, 25-400mg, 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, 20-30 0mg, 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, 9 0-200mg, 100-200mg, 125-200mg, 150-200mg, 1-100mg, 2-100mg, 5-100mg, 10-100mg, 15-100mg, 20-100mg, 25-100mg, 30-100mg, 40-100mg, 50-100mg,60-100mg, 70-100mg, 75-100mg, 80-100mg, 90-100mg; in some embodiments, the formulation specifications of the pharmaceutical composition include but are not limited to 1mg, 5mg, 10mg, 15mg, 20mg, 25mg, 30mg, 35mg, 40mg, 45mg, 50mg, 55mg, 65mg, 70mg, 75mg, 80mg, 85mg, 90mg, 95mg, 100mg, 110mg, 120mg, 125mg, 130mg, 140mg, 150mg, 160mg, 170mg, 180mg, 190mg, 200mg, 210mg, 220mg, 230mg, 240mg, 250mg, 300mg.

[0077] In some embodiments, the amount of active ingredient A in a unit dosage form of the pharmaceutical composition or pharmaceutical preparation of the present invention includes but is not limited to 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, 250-600 mg, 300-600 mg, 400-600 mg, 500-600 mg, 600-600 mg, 700-600 mg, 750-600 mg, 800-600 mg, 900-600 mg, 1000-600 mg, 2 ... 0mg, 1-500mg, 2-500mg, 3-500mg, 4-500mg, 5-500mg, 6-500mg, 10-500mg, 20-500mg, 25-500mg, 30-500mg, 40-500mg, 50-500 mg, 60-500mg, 70-500mg, 75-500mg, 80-500mg, 90-500mg, 100-500mg, 125-500mg, 150-500mg, 200-500mg, 250-500mg, 300-50 0mg, 400-500mg, 5-400mg, 10-400mg, 20-400mg, 25-400mg, 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, 100-30 0mg, 125-300mg, 150-300mg, 200-300mg, 250-300mg, 1-200mg, 2-200mg, 5-200mg, 10-200mg, 20-200mg, 25-200mg, 30-200mg,40-200 mg, 50-200 mg, 60-200 mg, 70-200 mg, 75-200 mg, 80-200 mg, 90-200 mg, 100-200 mg, 125-200 mg, 150-200 mg, 1-100 mg, 2-100 mg, 5-100 mg, 10-100 mg, 15-100 mg, 20-100 mg, 25-100 mg, 30-100 mg, 40-100 mg, 50-100 mg, 60-100 mg, 70-100 mg, 75-100 mg, 80-100 mg, 90-100 mg; in some embodiments The amount of active ingredient A in a unit preparation of the pharmaceutical composition or pharmaceutical preparation includes but is not limited to 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, 125 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.

[0078] The present invention relates to a kit, which may include a pharmaceutical composition or pharmaceutical preparation in a single-dose or multi-dose form, wherein the kit contains the active ingredient A in the pharmaceutical composition of the present invention in an amount including but not limited to 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, 125 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, and 300 mg.

[0079] Unless otherwise stated, the terms used in this specification and claims have the following meanings.

[0080] "Preparation specifications" refers to the weight of the main drug (active ingredient A) contained in each vial, tablet or other unit preparation.

[0081] 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 further 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.

[0082] "Halogen" refers to F, Cl, Br or I.

[0083] "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".

[0084] "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.

[0085] "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.

[0086] "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.

[0087] "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.

[0088] "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.

[0089] "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.

[0090] "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.

[0091] "Carbocyclyl" or "carbocycle" refers to a substituted or unsubstituted saturated or unsaturated aromatic or non-aromatic ring, which can be a 3-8 membered monocyclic ring, a 4-12 membered bicyclic ring, or a 10-15 membered tricyclic ring system, and the carbocyclyl can be attached to the aromatic or non-aromatic ring, which can be optionally a monocyclic ring, a bridged ring, or a spirocyclic ring. Non-limiting examples include cyclopropane, cyclobutane, cyclopentane, cyclohexane, cycloheptane, 1-cyclopentyl-1-alkenyl, 1-cyclopentyl-2-alkenyl, 1-cyclopentyl-3-alkenyl, cyclohexyl, 1-cyclohexyl-2-alkenyl, 1-cyclohexyl-3-alkenyl, cyclohexenyl, a benzene ring, a naphthalene ring, "Carbocyclyl" or "carbocycle" can be monovalent, divalent, trivalent, or tetravalent.

[0092] "Heterocyclyl" or "heterocycle" refers to a substituted or unsubstituted saturated or unsaturated aromatic or non-aromatic ring, which can be a 3-8 membered monocyclic ring, a 4-12 membered bicyclic ring or a 10-15 membered tricyclic ring system, and contains one or more (including but not limited to 2, 3, 4 or 5) heteroatoms selected from N, O or S. The N and S optionally substituted in the heterocyclyl ring can be oxidized to various oxidation states. The heterocyclic group can be connected to a heteroatom or a carbon atom, the heterocyclic group can be connected to an aromatic ring or a non-aromatic ring, and the heterocyclic group can be connected to a bridged ring or a spiro ring. Non-limiting examples include oxirane, aziridine, oxetanyl, azetidinyl, 1,3-dioxolanyl, 1,4-dioxolanyl, 1,3-dioxhexacyclyl, azepanyl, pyridyl, furyl, thienyl, pyranyl, N-alkylpyrrolyl, pyrimidinyl, pyrazinyl, pyridazinyl, imidazolyl, piperidinyl, morpholinyl, thiomorpholinyl, 1,3-dithianyl, dihydrofuranyl, dihydropyranyl, dithiolanyl, tetrahydrofuranyl, py ... furanyl, tetrahydropyrrolyl, tetrahydroimidazolyl, tetrahydrothiazolyl, tetrahydropyranyl, benzimidazolyl, benzopyridinyl, pyrrolopyridinyl, benzodihydrofuranyl, pyrrolyl, pyrazolyl, thiazolyl, oxazolyl, pyrazinyl, indazolyl, benzothiophenyl, benzofuranyl, benzopyrrolyl, benzimidazolyl, benzothiazolyl, benzoxazolyl, benzopyridinyl, benzopyrimidinyl, benzopyrazinyl, piperazinyl, azabicyclo[3.2.1]octyl, azabicyclo[5.2.0]nonyl, oxatricyclo[5.3.1.1]dodecyl, azaadamantyl, oxaspiro[3.3]heptanyl, "Heterocyclyl" or "heterocycle" can be monovalent, divalent, trivalent or tetravalent.

[0093] "Aryl" or "aromatic ring" refers to a substituted or unsubstituted aromatic hydrocarbon group having a single ring or a fused ring, wherein the number of ring atoms in the aromatic ring includes, but is not limited to, 6 to 18, 6 to 12, or 6 to 10 carbon atoms. The aryl ring may be fused to a saturated or unsaturated carbocyclic or heterocyclic ring, wherein the ring connected to the parent structure is the aryl ring, non-limiting examples of which include benzene ring, naphthalene ring, "Aryl" or "aromatic ring" can be monovalent, divalent, trivalent or tetravalent. When divalent, trivalent or tetravalent, the point of attachment is on the aryl ring.

[0094] "Heteroaryl" or "heteroaromatic ring" refers to a substituted or unsubstituted aromatic hydrocarbon group containing 1 to 5 heteroatoms or a group containing heteroatoms (including but not limited to N, O or S(=O)n, n is 0, 1, 2), and the number of ring atoms in the heteroaromatic ring is but not limited to 5 to 15, 5 to 10 or 5 to 6. Non-limiting examples of heteroaryl include but are not limited to pyridyl, furyl, thienyl, pyridyl, pyranyl, N-alkylpyrrolyl, pyrimidinyl, pyrazinyl, pyridazinyl, imidazolyl, benzopyrazolyl, benzimidazolyl, benzopyridinyl, pyrrolopyridinyl and the like. The heteroaryl ring may be fused to a saturated or unsaturated carbocyclic ring or heterocyclic ring, wherein the ring connected to the parent structure is a heteroaryl ring, and non-limiting examples include When heteroaryl appears in this document, its definition is consistent with this definition. Heteroaryl can be monovalent, divalent, trivalent or tetravalent. When it is divalent, trivalent or tetravalent, the attachment point is located on the heteroaryl ring.

[0095] "5-membered ring and 5-membered heteroaromatic ring" refers to a 5-membered fused heteroaromatic ring, at least one of the two fused rings contains one or more heteroatoms (including but not limited to O, S or N), and the entire group is aromatic. Non-limiting examples include pyrrolopyrrole ring, pyrazolopyrrole ring, pyrazolopyrazole ring, pyrrolofuran ring, pyrazolofuran ring, pyrrolothiophene ring, and pyrazolothiophene ring.

[0096] "5-membered and 6-membered heteroaromatic ring" refers to a 5-membered and 6-membered fused heteroaromatic ring, at least one of the two fused rings contains one or more heteroatoms (including but not limited to O, S or N), and the entire group is aromatic. Non-limiting examples include benzo 5-membered heteroaromatic group, 6-membered heteroaromatic ring and 5-membered heteroaromatic ring.

[0097] "Substituted" or "substituted" refers to substitution by one or more (including but not limited to 2, 3, 4 or 5) substituents, including but not limited to H, F, Cl, Br, I, alkyl, cycloalkyl, alkoxy, haloalkyl, thiol, hydroxyl, nitro, mercapto, amino, cyano, isocyano, aryl, heteroaryl, heterocyclic, bridged, spiro, cycloalkyl, hydroxyalkyl, =O, carbonyl, aldehyde, carboxylic acid, formate, -(CH2), m -C(=O)-R a 、-O-(CH2) m -C(=O)-R a 、-(CH2) m -C(=O)-NR b R c 、-(CH2) m S(=O) n R a 、-(CH2) m -alkenyl-R a , ORd or -(CH2) m -alkynyl-R a (wherein m and n are 0, 1 or 2), arylthio, thiocarbonyl, silyl or -NR b R c etc., where R b With R c R is independently selected from the group consisting of H, hydroxy, amino, carbonyl, alkyl, alkoxy, cycloalkyl, heterocyclyl, aryl, heteroaryl, sulfonyl, trifluoromethanesulfonyl, and optionally, b With R c It can form a five- or six-membered cycloalkyl or heterocyclic group. a With R d Each is independently selected from aryl, heteroaryl, alkyl, alkoxy, cycloalkyl, heterocyclyl, carbonyl, ester, bridged ring, spiro ring or paracyclic group.

[0098] “Containing 1 to 5 heteroatoms selected from O, S, and N” means containing 1, 2, 3, 4, or 5 heteroatoms selected from O, S, and N.

[0099] "Substituted with 0 to X substituents" means substituted with 0, 1, 2, 3, ..., X substituents, where X is selected from any integer between 1 and 10. For example, "substituted with 0 to 4 substituents" means substituted with 0, 1, 2, 3, or 4 substituents. For example, "substituted with 0 to 5 substituents" means substituted with 0, 1, 2, 3, 4, or 5 substituents. For example, "the heterobridged ring is optionally substituted with 1 to 4 substituents selected from H or F" means that the heterobridged ring is optionally substituted with 1, 1, 2, 3, or 4 substituents selected from H or F.

[0100] XY-membered rings (where X is selected from an integer less than Y and greater than 3, and Y is selected from any integer between 4 and 12) include rings with X+1, X+2, X+3, X+4, and so on. Rings include heterocycles, carbocycles, aromatic rings, aryl groups, heteroaryl groups, cycloalkyl groups, heteromonocycles, heteroplexes, heterospirocycles, and 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 heteroplexe.

[0101] "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.

[0102] "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.

[0103] "Pharmaceutical composition" refers to a mixture of one or more compounds of the present invention, or stereoisomers, tautomers, deuterated forms, solvates, prodrugs, metabolites, pharmaceutically acceptable salts or cocrystals thereof, and other chemical components, wherein "other chemical components" refers to pharmaceutically acceptable carriers, excipients and / or one or more other therapeutic agents.

[0104] "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.

[0105] "Excipient" refers to an inert substance added to a pharmaceutical composition to facilitate administration of a compound. Non-limiting examples include calcium carbonate, calcium phosphate, sugars, starches, cellulose derivatives (including microcrystalline cellulose), gelatin, vegetable oils, polyethylene glycols, diluents, granulating agents, lubricants, binders, and disintegrants.

[0106] "Prodrugs" refer to compounds of the present invention that can be converted into biologically active compounds through in vivo metabolism. Prodrugs of the present invention are prepared by modifying amino or carboxyl groups in compounds of the present invention. These modifications can be removed by conventional manipulation or in vivo to yield the parent compound. When the prodrugs of the present invention are administered to a mammalian subject, the prodrugs are cleaved to form free amino or carboxyl groups.

[0107] A "cocrystal" is a crystal formed by the active pharmaceutical ingredient (API) and cocrystal former (CCF) bound together by hydrogen bonds or other non-covalent bonds. Both the API and CCF are solid in their pure form at room temperature, and the components exist in a fixed stoichiometric ratio. A cocrystal is a multi-component crystal, encompassing both binary cocrystals formed between two neutral solids and multi-component cocrystals formed between a neutral solid and a salt or solvate.

[0108] "Animal" is meant to include mammals, such as humans, companion animals, zoo animals, and livestock, preferably humans, horses, or dogs.

[0109] "Stereoisomers" refer to isomers resulting from different spatial arrangements of atoms in a molecule, including cis-trans isomers, enantiomers, and conformational isomers.

[0110] "Tautomers" refer to functional group isomers produced by the rapid movement of an atom in a molecule between two positions, such as keto-enol isomers and amide-imino alcohol isomers.

[0111] “IC 50 "It is the concentration of a drug or inhibitor required to inhibit a specified biological process (or a component of the process such as an enzyme, receptor, cell, etc.) by half. BRIEF DESCRIPTION OF THE DRAWINGS

[0112] FIG1 is an X-ray powder diffraction pattern of the tris(hydroxymethyl)aminomethane salt form A of compound 6.

[0113] FIG2 is a differential scanning calorimetry curve and a thermogravimetric analysis spectrum of the tris(hydroxymethyl)aminomethane salt Form A of Compound 6. DETAILED DESCRIPTION

[0114] The technical solutions of the present invention are described in detail below with reference to the accompanying drawings and embodiments, but the protection scope of the present invention includes but is not limited to them.

[0115] The compounds used in the reactions described herein were prepared using 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" were obtained from reputable commercial sources, including suppliers such as Titan Technology, Anage Chemical, Shanghai Demo, Chengdu Kelon Chemical, Shaoyuan Chemical Technology, Nanjing Yaoshi, WuXi AppTec, and J&K Technology.

[0116] The structures of the compounds were determined by nuclear magnetic resonance (NMR) or / and mass spectrometry (MS). NMR shifts (δ) were expressed in 10 -6 The unit of (ppm) is given. 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).

[0117] MS was determined using (Agilent 6120B (ESI) and Agilent 6120B (APCI));

[0118] HPLC determination was performed using an Agilent 1260DAD high-pressure liquid chromatograph (Zorbax SB-C18 100 × 4.6 mm, 3.5 μM);

[0119] Thin layer chromatography silica gel plate using Yantai Huanghai HSGF 254 or Qingdao GF 254 Silica gel plates, the specifications of silica gel plates used in thin layer chromatography (TLC) are 0.15mm-0.20mm, and the specifications used for thin layer chromatography separation and purification products are 0.4mm-0.5mm;

[0120] Column chromatography generally uses Yantai Huanghai silica gel 200-300 mesh silica gel as the carrier;

[0121] Boc: tert-butyloxycarbonyl; Ts: p-toluenesulfonyl; Cbz: benzyloxycarbonyl; TMS: trimethylsilyl.

[0122] Example 1: Preparation of Compound 6

[0123] first step:

[0124] 6a (5.0 g, 26.04 mmol) was added to anhydrous ethanol (3.0 mL). While stirring thoroughly at 0°C, dry hydrogen chloride gas was introduced into the reaction solution until saturated. The mixture was stirred at 0°C for 10 hours. Diethyl ether (40 mL) was then added to the reaction solution, filtered, and the resulting solid was washed with diethyl ether (100 mL x 2). The solid was dried to afford 6b (5.0 g, 72% yield).

[0125] 1 H NMR (400MHz, DMSO-d6) δ12.04(s,2H),7.61(d,1H),7.51–7.43(m,2H),4.46(q,2H),4.14(s,2H),1.26(t,3H).

[0126] 19 F NMR (400MHz, DMSO-d6) δ111.30.

[0127] Step 2:

[0128] 6b (5.0 g, 19.30 mmol) was added to anhydrous ethanol (40 mL) and stirred at room temperature (15°C) for 48 hours. The reaction solution was then filtered to obtain a filtrate, which was then spin-dried to dryness and then slurried with a mixed solvent of petroleum ether and ethyl acetate (100 mL) (petroleum ether / ethyl acetate (v / v) = 10:1). The mixture was filtered and the filtrate was spin-dried to obtain 6c (4.0 g, 66% yield).

[0129] 1 H NMR (400MHz, CDCl3)7.45(t,1H),7.23–7.14(m,2H),3.50(q,6H),3.07(s,2H),1.16(t,9H).

[0130] Step 3:

[0131] 1d (500 mg, 2.06 mmol) and 6c (1.0 g, 3.19 mmol) were added to glacial acetic acid (20 mL) and stirred at 60°C for 10 hours. The reaction system was cooled to room temperature and slowly added to a saturated aqueous solution of sodium bicarbonate (200 mL) to quench the mixture. The mixture was then extracted with ethyl acetate (50 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, and filtered. The filtrate was then spin-dried to dryness, and the residue was purified by silica gel column chromatography to afford 6e (430 mg, 47% yield).

[0132] LCMS m / z=439.0[M+1] + .

[0133] Step 4:

[0134] 6e (400 mg, 0.98 mmol), pinacol diboronate (262 mg, 1.0 mmol), Pd(dppf)Cl2·DCM (81 mg, 0.1 mmol), and potassium acetate (196 mg, 2.0 mmol) were added to 1,4-dioxane (20 mL). The reaction flask was replaced with nitrogen and stirred at 100°C for 5 hours. The reaction solution was cooled to room temperature, added with water (100 mL), and extracted with ethyl acetate (50 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, and filtered. The filtrate was spin-dried to dryness, and the residue was purified by silica gel column chromatography to afford 6f (340 mg, 71% yield).

[0135] LCMS m / z=487.2[M+1] + .

[0136] Step 5: 6f (340 mg, 0.70 mmol), 6g (220 mg, 0.84 mmol), Pd(dppf)Cl2·DCM (83 mg, 0.1 mmol), and potassium carbonate (290 mg, 2.1 mmol) were added to a mixture of 1,4-dioxane (10 mL) and water (2 mL). The reaction flask was replaced with nitrogen and stirred at 100°C for 5 hours. The reaction solution was cooled to room temperature, water (50 mL) was added, and the mixture was extracted with ethyl acetate (30 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, and filtered. The filtrate was spin-dried to dryness, and the residue was purified by silica gel column chromatography to afford 6h (200 mg, 48% yield).

[0137] LCMS m / z=587.1[M+1] + .

[0138] Step 6:

[0139] 6h (100 mg, 0.17 mmol) and 1g (120 mg, 0.85 mmol) were added to a mixed solution of acetonitrile (5 mL) and water (1 mL) and stirred at room temperature (12°C) for 10 hours. The reaction solution was then adjusted to pH 6 with 1M hydrochloric acid and stirred until a white solid precipitated. The solid was filtered and washed with water. The solid was dissolved in dichloromethane and dried over anhydrous sodium sulfate, filtered, and the organic phase was spin-dried. The residue was separated by HPLC to obtain the trifluoroacetic acid salt of compound 6, which was then purified by silica gel column chromatography to obtain compound 6 (40 mg, 41% yield).

[0140] LCMS m / z=573.1[M+1] + .

[0141] 1 H NMR(400MHz,CD3OD)δ12.90(br,1H),7.91(d,1H),7.88–7.80(m,4H),7.79–7.68(m,2H)7.64(d,1H),7.39(t,1H),6.92(d,1H ),5.61(s,2H),5.06–4.98(m,1H),4.60(dd,1H),4.54–4.44(m,2H),4.43–4.28(m,3H),2.73–2.61(m,1H),2.40–2.27(m,1H).

[0142] Preparation of tris(hydroxymethyl)aminomethane salt of compound 6:

[0143] Compound 6 (1.9 g, 3.32 mmol) was dissolved in 50 mL of n-propanol, and a solution of tris(hydroxymethylaminomethane) (0.40 g, 3.32 mmol) in water (5 mL) was added. The mixture was reacted at room temperature for 16 h. The solid was filtered and dried to obtain the Tris salt of compound 6 (i.e., tris(hydroxymethylaminomethane) salt of compound 6) (1.8 g, yield 78%).

[0144] 1 H NMR(400MHz,DMSO-d6)δ7.94-7.88(m,1H),7.88-7.80(m,3H),7.79-7.69(m,2H),7.66-7.60(m,1H),7.47(s,1H),7.40-7.32(m,1H ),6.95-6.87(m,1H),5.61(s,2H),5.02-4.93(m,1H),4.57-4.21(m,6H),3.50-3.36(m,6H),2.71-2.60(m,1H),2.38–2.27(m,1H).

[0145] LCMS m / z=573.1[M+1] + .

[0146] The Tris salt of compound 6 was subjected to XRD, TGA and DSC tests. According to the XRD test, the crystalline form was named Form A of the Tris salt of compound 6.

[0147] Table B-1 XRD test parameters

[0148] 2. DSC test of compounds

[0149] The DSC spectrum was collected on a TA Instruments discovery DSC 2500 differential scanning calorimeter. The test parameters of the tris (hydroxymethyl)aminomethane salt of compound 6 are shown in Table B-2.

[0150] Table B-2 DSC test parameters

[0151] 3. TGA test of compounds

[0152] The TGA spectra were collected on TA Instruments discovery TGA 55 and TGA 550 thermogravimetric analyzers. The test parameters of the tris (hydroxymethyl)aminomethane salt of compound 6 are shown in Table B-3.

[0153] Table B-3 TGA test parameters

[0154] The specific peaks of the XRD test of the Tris salt of compound 6 are shown in Table B-4, and the XRD pattern is shown in Figure 1.

[0155] Table B-4

[0156] The Tris salt of compound 6 was tested at 40°C + 75% RH and 30°C + 60% RH, and the purity (expressed as a percentage) was determined by HPLC.

[0157] The preparation method of the test solution and the HPLC purity test conditions are shown in Tables B-5 and B-6;

[0158] Table B-5 Preparation method of test solution

[0159] Table B-6 HPLC purity test conditions

[0160] Table B-7 Chemical stability of compound 6 (purity determined by HPLC)

[0161] Conclusion: The tris(hydroxymethyl)aminomethane salt form A of compound 6 has good chemical stability.

[0162] Example 2: Preparation of Compound 7

[0163] first step:

[0164] 7a (19.6 g, 100.0 mmol) was dissolved in 33% ethanolic hydrochloric acid (45 mL) in an ice bath and allowed to react overnight. Anhydrous ether (100 mL) was added to the reaction system at room temperature, stirred for 5 minutes, and filtered. The filter cake was washed with ether (20 mL x 2) and dried to afford 7b (22.0 g, 79.4% yield) as a white solid.

[0165] LCMS m / z=242.1[M+1] + .

[0166] Step 2:

[0167] Substrate 7b (1.4 g, 5.0 mmol) was dissolved in glacial acetic acid (50 mL) at room temperature, and 1d (1.2 g, 5.0 mmol) was added. After addition, the reaction system was heated to 45°C under nitrogen overnight. The solvent was removed by distillation under reduced pressure to obtain a crude product, which was then purified by silica gel column chromatography to afford 7c (740.0 mg, 35.2% yield) as a white solid.

[0168] LCMS m / z=421.0[M+1] + .

[0169] Step 3:

[0170] At room temperature, 7c (740.0 mg, 1.8 mmol) was dissolved in 1,4-dioxane (30 mL). Diboronic acid pinacol ester (820.0 mg, 2.7 mmol), Pd(dppf)Cl2 (147.0 mg, 0.2 mmol), and potassium acetate (519.0 mg, 5.3 mmol) were added sequentially. The mixture was heated to 100°C under nitrogen and allowed to react overnight. The reaction mixture was cooled to room temperature, filtered, and the filter cake was washed with dichloromethane (10 mL x 2). The filtrate was collected and concentrated to obtain the crude product, which was purified by silica gel column chromatography to afford 7d (680.0 mg, 82.4% yield) as a yellow solid.

[0171] LCMS m / z=469.2[M+1] + .

[0172] Step 4:

[0173] At room temperature, 3a (130.0 mg, 0.5 mmol) was dissolved in a mixture of 1,4-dioxane (10 mL) and water (1 mL). 7d (193.0 mg, 0.4 mmol), Pd(dppf)Cl2 (34.0 mg, 0.04 mmol), and cesium carbonate (261.0 mg, 0.8 mmol) were then added sequentially. The mixture was heated to 100°C under nitrogen and allowed to react overnight. The reaction mixture was cooled to room temperature, filtered, and the filter cake was washed with dichloromethane (10 mL x 2). The filtrate was collected and concentrated to obtain the crude product, which was then purified by silica gel column chromatography to afford 7e (109.0 mg, 46.6% yield) as a white solid.

[0174] LCMS m / z=569.3[M+1] + .

[0175] Step 5:

[0176] At room temperature, 7e (109.0 mg, 0.2 mmol) was dissolved in acetonitrile (5 mL) and water (1 mL). 1 g (CAS: 5807-14-7) (80.0 mg, 0.6 mmol) was added and allowed to react at room temperature for 24 hours. The acetonitrile was removed by distillation under reduced pressure, and the crude product was dissolved in water (5 mL). The pH was adjusted to 6 with 1N hydrochloric acid. The aqueous phase was extracted with ethyl acetate (15 mL x 3). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product. The crude product was purified by silica gel column chromatography to obtain compound 7 (26.0 mg, 24.5% yield) as a white solid.

[0177] LCMS m / z=555.2[M+1] + .

[0178] 1 H NMR(400MHz,DMSO-d6)δ7.98(d,2H),7.93–7.88(m,1H),7.82(t,1H),7.78–7.68(m,3H),7.57(d,1H),7.38(d ,2H),6.87(d,1H),5.60(s,2H),4.94–4.86(m,1H),4.54–4.30(m,6H),2.65–2.56(m,1H),2.33–2.24(m,1H).

[0179] Example 3: Preparation of Compound 8

[0180] first step:

[0181] 8a (10.0 g, 43 mmol) and a 2 M borane solution in tetrahydrofuran (26 mL, 51.6 mmol) were added to tetrahydrofuran (40 mL) and stirred at room temperature for 30 hours. The reaction solution was slowly added to 0.5 N hydrochloric acid (100 mL) to quench the mixture, followed by extraction with ethyl acetate (80 mL × 3). The organic phase was dried over anhydrous sodium sulfate and filtered. The liquid was concentrated to dryness, and the residue was purified by silica gel column chromatography to obtain 8b (9.4 g, 99% yield).

[0182] LCMS m / z=201.0[M-17] + .

[0183] Step 2:

[0184] 8b (2.18 g, 10 mmol) and triethylamine (2.0 g, 20 mmol) were added to dry dichloromethane (20 mL) and stirred at 0°C for 10 minutes. Methanesulfonyl chloride (1.38 g, 12 mmol) was then slowly added to the reaction system, and the reaction was stirred at 0°C for another 30 minutes. Water (20 mL) was then added to the reaction solution, and the mixture was extracted with dichloromethane (50 mL x 3). The organic phase was dried over anhydrous sodium sulfate and filtered. The filtrate was then concentrated under reduced pressure to afford 8c (3.0 g, 99% yield), which was used directly in the next step without further purification.

[0185] Step 3:

[0186] 8c (3.0 g, 10 mmol) and sodium cyanide (440 mg, 9 mmol) were added to DMF (15 mL) and stirred at room temperature for 40 hours. Ethyl acetate (300 mL) was then added to the reaction system, which was then washed with water (50 mL × 3). Aqueous sodium hypochlorite (50 mL) was added to the aqueous phase and stirred for 5 hours. The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to dryness. 8d (1.5 g, 66% yield) was obtained by silica gel column chromatography.

[0187] Step 4:

[0188] 8d (1.5 g, 6.6 mmol) was added to ethanol (5 mL) and stirred at room temperature. Dry hydrogen chloride gas was then passed through the reaction solution for 1 hour, followed by further stirring for 10 hours. Diethyl ether (10 mL) was added to the reaction solution and stirred until a solid precipitated. The solid was filtered, and the filter cake was washed with diethyl ether and dried to afford 8e (1.5 g, 73% yield).

[0189] Step 5:

[0190] 8e (300 mg, 0.96 mmol) and 1d (250 mg, 1.0 mmol) were added to glacial acetic acid (10 mL). After nitrogen replacement, the mixture was stirred at 40°C for 10 hours. The reaction mixture was then quenched by adding saturated aqueous sodium bicarbonate solution and extracted with ethyl acetate (30 mL x 3). The organic phase was dried over anhydrous sodium sulfate and filtered. The liquid was concentrated to dryness, and the residue was purified by silica gel column chromatography to afford 8f (100 mg, 25% yield).

[0191] LCMS m / z=453.0[M+1] + .

[0192] Step 6:

[0193] 8f (100 mg, 0.22 mmol), bis(pinacol)boronate (69 mg, 0.26 mmol), Pd(dppf)Cl2 (18 mg, 0.022 mmol), and potassium acetate (45 mg, 0.44 mmol) were added to 1,4-dioxane (8 mL). After nitrogen replacement, the mixture was stirred at 100°C for 12 hours. The reaction solution was then cooled to room temperature, concentrated to dryness, and purified by silica gel column chromatography to yield 8 g (70 mg, 63% yield).

[0194] LCMS m / z=501.2[M+1] + .

[0195] Step 7:

[0196] 8g (70mg, 0.14mmol), 3a (50mg, 0.2mmol), Pd(dppf)Cl2 dichloromethane complex (18mg, 0.02mmol), and potassium carbonate (55mg, 0.4mmol) were added to a mixture of 1,4-dioxane (5mL) and water (1mL). After replacing the solvent with nitrogen, the mixture was stirred at 100°C for 6 hours. The reaction solution was cooled to room temperature, quenched with saturated aqueous ammonium chloride (20mL), and extracted with ethyl acetate (20mL x 3). The organic phase was dried over anhydrous sodium sulfate, filtered and dried, and purified by silica gel column chromatography to obtain 8h (60mg, 71% yield).

[0197] LCMS m / z=601.3[M+1] + .

[0198] Step 8:

[0199] 8h (60 mg, 0.1 mmol) and 1g (CAS: 5807-14-7) (70 mg, 0.5 mmol) were added to a mixture of acetonitrile (5 mL) and water (1.0 mL) and stirred at 50°C for 5 hours. The mixture was cooled to room temperature, adjusted to pH 6 with 1N hydrochloric acid, and extracted with ethyl acetate (20 mL x 3). The organic phases were combined and dried over anhydrous sodium sulfate. After filtration, the liquid was concentrated to dryness under reduced pressure. The residue was purified by silica gel chromatography to obtain compound 8 (10 mg, 17% yield).

[0200] LCMS m / z=587.3[M+1] + .

[0201] 1 H NMR(400MHz,DMSO-d6)δ12.89(s,1H),7.95–7.80(m,3H),7.77–7.66(m,2H),7.26–7.16(m,3H),6.94(d,1H),6.51(s,2H), 5.06–4.97(m,1H),4.61(m,1H),4.54–4.45(m,2H),4.39–4.22(m,3H),2.74–2.65(m,1H),2.38–2.30(m,1H),2.22(s,3H).

[0202] Example 4: Preparation of Compound 9

[0203] first step:

[0204] Raw materials 9a (120 mg, 0.87 mmol) and 2b (300 mg, 0.87 mmol) were added to a reaction flask and dissolved in acetonitrile (10 mL). Anhydrous potassium carbonate (250 mg, 1.80 mmol) was added and reacted at 40°C for 4 h. After cooling to room temperature, the solid was filtered and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography to obtain product 9b as a yellow solid (247 mg, yield 71.8%).

[0205] LCMS m / z=395.0[M+H] + .

[0206] Step 2:

[0207] 9b (40 mg, 0.10 mmol) and 9c (30 mg, 0.20 mmol), anhydrous potassium carbonate (40 mg, 0.20 mmol), and Pd(dppf)Cl2 (15 mg, 0.01 mmol) were added sequentially to a reaction flask and dissolved in a mixed solution of 1,4-dioxane (2 mL) and water (0.4 mL). The atmosphere was replaced with nitrogen and stirred at 90°C for 5 h. The mixture was cooled to room temperature and filtered. The filtrate was concentrated under reduced pressure and the residue was purified by silica gel column chromatography to give product 9d as a yellow solid (22 mg, 48.3% yield).

[0208] LCMS m / z=456.1[M+H] + .

[0209] Step 3:

[0210] 9d (22 mg, 0.05 mmol) and 9f (15 mg, 0.09 mmol) were added to a reaction flask and dissolved in tetrahydrofuran solution (1 mL). Potassium tert-butoxide (10 mg, 0.07 mmol) was added under nitrogen. After stirring at room temperature for 1 h, the reaction was quenched with aqueous ammonium chloride (1 mL). The mixture was extracted with ethyl acetate (2 mL × 3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was separated and purified by silica gel column chromatography to give 9h as a yellow solid (20 mg, 71.1% yield).

[0211] LCMS m / z=587.1[M+H] + .

[0212] Step 4: Dissolve 9h (20 mg, 0.03 mmol) in acetonitrile (2 mL) and water (0.5 mL) at room temperature, then add 1 g (CAS: 5807-14-7) (40 mg, 0.28 mmol) and continue the reaction for 24 h. Adjust the pH to 6 with 1N hydrochloric acid, then extract with dichloromethane:methanol = 10:1 (2 mL x 3). The combined organic phases are dried over anhydrous sodium sulfate, filtered, and concentrated. The residue is purified by silica gel chromatography to afford compound 9 (5 mg, 25.7% yield).

[0213] LCMS m / z=573.1[M+H] + .

[0214] 1H NMR (400MHz, DMSO-d6) δ13.14–12.78(m,1H),8.62(d,1H),7.98–7.68(m,6H),7.57(d,1H),7.08(d,1H),5.68–5. 50(m,4H),5.04(d,1H),4.70(dd,1H),4.58(d,1H),4.48(dd,1H),4.33(dd,1H),2.74–2.63(m,1H),2.33(dd,1H).

[0215] Example 5: Preparation of Compound 10

[0216] first step:

[0217] 10a (10.0 g, 52.1 mmol) was added to a reaction flask, and 33% hydrogen chloride-ethanol solution was introduced under an ice bath. The temperature was gradually raised to room temperature and the reaction was allowed to proceed overnight. A large amount of white solid precipitated. Ether was added and stirred for 0.5 h. The solid was filtered, washed with ether, and dried to obtain 10b as a white solid (9.3 g, 60.6% yield).

[0218] Step 2:

[0219] 10b (500 mg, 2.06 mmol) and 1d (620 mg, 2.10 mmol) were dissolved in glacial acetic acid (20 mL), the atmosphere was replaced with nitrogen, and the mixture was stirred at 45°C for 12 h. The mixture was then cooled to room temperature and concentrated under reduced pressure. The residue was purified by silica gel column chromatography to afford the desired product 10c as a yellow solid (350 mg, 39.9% yield).

[0220] LCMS m / z=439.0[M+H] + .

[0221] Step 3:

[0222] 10c (350 mg, 0.79 mmol), pinacol diboronate (305 mg, 1.20 mmol), potassium acetate (235 mg, 2.40 mmol), and Pd(dppf)Cl2 (65 mg, 0.08 mmol) were added to a reaction flask in this order and dissolved in 1,4-dioxane solution (10 mL). The atmosphere was replaced with nitrogen and stirred at 90°C for 5 h. The mixture was cooled to room temperature, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product 10d as a yellow oil, which was used directly in the next reaction without further purification.

[0223] Step 4:

[0224] Crude products 10d and 4c (344 mg, 1.01 mmol), anhydrous potassium carbonate (230 mg, 1.60 mmol), and Pd(dppf)Cl2 (65 mg, 0.08 mmol) were added sequentially to a reaction flask and dissolved in a mixed solution of 1,4-dioxane (10 mL) and water (2 mL). The atmosphere was replaced with nitrogen and stirred at 90°C for 5 h. The mixture was cooled to room temperature, the solid was filtered, and the filtrate was concentrated under reduced pressure and purified on a silica gel column to obtain product 10e as a yellow solid (350 mg, two-step yield 71.2%).

[0225] LCMS m / z=623.1[M+H] + .

[0226] Step 5:

[0227] 10e (350 mg, 0.70 mmol) was dissolved in acetonitrile (5 mL) and water (1 mL) at room temperature, followed by the addition of lithium hydroxide (126.0 mg, 3.01 mmol). The reaction was continued for 24 h. The pH was adjusted to 6 with 1N hydrochloric acid, and the mixture was extracted with a 10:1 dichloromethane:methanol mixture (20 mL x 3). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography to afford compound 10 as a yellow solid (100.0 mg, 29.3% yield).

[0228] LCMS m / z=609.2[M+H] + .

[0229] 1 H NMR(400MHz,DMSO-d6)δ7.73(br,1H),7.62-7.54(m,4H),7.42-7.33(m,2H),7.22-7.18(m,2H),7.01-6.94(m,2H),5.04- 4.96(m,1H),4.62-4.53(m,1H),4.52-4.42(m,2H),4.40-4.27(m,3H),2.73-2.61(m,1H),2.39-2.27(m,1H),2.09(s,3H).

[0230] Example 6: Preparation of Compound 11

[0231] first step:

[0232] At room temperature, 4c (91.0 mg, 0.26 mmol) was dissolved in a mixture of 1,4-dioxane (5 mL) and water (0.5 mL). 7d (103.0 mg, 0.22 mmol), Pd(dppf)Cl2 (18.0 mg, 0.02 mmol), and cesium carbonate (215.0 mg, 0.66 mmol) were then added sequentially. The mixture was heated to 100°C under nitrogen and allowed to react overnight. The reaction mixture was cooled to room temperature, filtered, and the filter cake was washed with dichloromethane (10 mL x 2). The filtrate was collected and concentrated to obtain the crude product, which was then purified by silica gel column chromatography to afford 11a (110.0 mg, 82.7% yield) as a white solid.

[0233] LCMS m / z=605.3[M+1] + .

[0234] Step 2:

[0235] Substrate 11a (110.0 mg, 0.18 mmol) was dissolved in a mixture of acetonitrile (5 mL) and water (1 mL) at room temperature. Then, 1 g (CAS: 5807-14-7)(4.4.0)-5-decene (23.0 mg, 0.54 mmol) was added. The mixture was allowed to react at room temperature for 24 hours. The acetonitrile was removed by distillation under reduced pressure, and the crude product was dissolved in water (5 mL). The pH was adjusted to 6 with 1N hydrochloric acid. The aqueous phase was extracted with ethyl acetate (15 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product. The crude product was purified by silica gel column chromatography to afford Compound 11 (25.0 mg, 23.4% yield) as a white solid.

[0236] LCMS m / z=591.2[M+1] + .

[0237] 1 H NMR(400MHz, DMSO-d6)δ12.90(brs,1H),7.82(s,1H),7.74–7.67(m,2H),7.62–7.52(m,2H),7.42–7.37(m,2H),7.36–7.32(dd,1H),7.16–7.10(m, 1H),6.95(d,2H),4.97–4.87(m,1H),4.60–4.52(m,1H),4.51–4.40(m,2H ),4.38–4.28(m,3H),2.68–2.56(m,1H),2.35–2.23(m,1H),2.06(s,3H).

[0238] Example 7: Preparation of Compound 18

[0239] first step:

[0240] Under ice, 18a (278 mg, 1.32 mmol) was added to a solution of 18b (200 mg, 1.32 mmol) and potassium tert-butoxide (221.8 mg, 1.98 mmol) in tetrahydrofuran (6 ml) and allowed to react at room temperature for 3 hours. The insoluble material was removed by filtration, washed once with saturated aqueous ammonium chloride (6 ml), dried over anhydrous sodium sulfate, and concentrated to dryness. The residue was purified by silica gel column chromatography to afford the desired product 18c (250 mg, 58.5% yield).

[0241] Step 2:

[0242] Crude products 10d and 18c (178 mg, 0.55 mmol), anhydrous potassium carbonate (115 mg, 0.80 mmol), and Pd(dppf)Cl2 (32.5 mg, 0.04 mmol) were added sequentially to a reaction flask and dissolved in a mixed solution of 1,4-dioxane (5 mL) and water (1 mL). The atmosphere was replaced with nitrogen and stirred at 90°C for 5 h. The solid was removed by filtration, and the filtrate was concentrated under reduced pressure. The residue was separated and purified by silica gel column chromatography to obtain product 18b as a yellow solid (140 mg, two-step yield 42.1%).

[0243] LCMS m / z=605.2[M+H] + .

[0244] Step 3:

[0245] 18b (80 mg, 0.13 mmol) was dissolved in acetonitrile (3 mL) and water (0.6 mL) at room temperature, followed by the addition of 1 g (CAS: 5807-14-7) (54.2 mg, 0.39 mmol). The reaction was continued for 24 h. The pH was adjusted to 6 with 1N hydrochloric acid, and the mixture was extracted with a 10:1 dichloromethane:methanol mixture (20 mL x 3). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography to afford compound 18 as a white solid (7 mg, 9.1% yield).

[0246] LCMS m / z=591.1[M+H] + .

[0247] 1H NMR(400MHz,DMSO-d6)δ7.96-7.91(m,1H),7.86-7.72(m,6H),7.70-7.65(m,1H),7.29(t,1H),5.70(s,2H),5.0 5-4.97(m,1H),4.64-4.56(m,1H),4.52-4.45(m,2H),4.41-4.28(m,3H),2.70-2.64(m,1H),2.35-2.29(m,1H).

[0248] Example 8: Preparation of Compound 19

[0249] first step:

[0250] Under ice, 18a (300 mg, 1.43 mmol) was added to a solution of 19b (285.7 mg, 1.43 mmol) and potassium tert-butoxide (240 mg, 2.15 mmol) in tetrahydrofuran (10 ml), followed by stirring for 3 hours. The insoluble material was removed by filtration, and the liquid was washed with an equal volume of water, dried over anhydrous sodium sulfate, and concentrated to dryness. The residue was purified by silica gel column chromatography to afford the desired product 19c (250 mg, 57.1% yield).

[0251] Step 2:

[0252] Crude products 10d and 19c (168.3 mg, 0.55 mmol), anhydrous potassium carbonate (115 mg, 0.80 mmol), and Pd(dppf)Cl2 (32.5 mg, 0.04 mmol) were added sequentially to a reaction flask and dissolved in a mixed solution of 1,4-dioxane (5 mL) and water (1 mL). The atmosphere was replaced with nitrogen and stirred at 90°C for 5 h. The mixture was cooled to room temperature, the solid was filtered, and the filtrate was concentrated under reduced pressure and purified on a silica gel column to give product 19d as a yellow solid (180 mg, two-step yield 67%).

[0253] LCMS m / z=587.1[M+H] + .

[0254] Step 3:

[0255] At room temperature, 19d (80 mg, 0.14 mmol) was dissolved in acetonitrile (3 mL) and water (0.6 mL). Then, 1 g (CAS: 5807-14-7) (56.9 mg, 0.41 mmol) was added and the reaction continued for 24 h. The pH was adjusted to 6 with 1N hydrochloric acid, and the mixture was extracted with a 10:1 dichloromethane:methanol mixture (20 mL x 3). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography to afford compound 19 as a white solid (40 mg, 50% yield).

[0256] LCMS m / z=573.1[M+H] + .

[0257] 1 H NMR(400MHz,DMSO-d6)δ7.90-7.77(m,6H),7.71(d,2H),7.68-7.64(m,1H),7.38(t,1H),5.67(s,2H),5.04- 4.96(m,1H),4.63-4.55(m,1H),4.53-4.44(m,2H),4.42-4.28(m,3H),2.72-2.62(m,1H),2.38-2.27(m,1H).

[0258] Example 9: Preparation of Compound 20

[0259] first step:

[0260] 2,6-Dichloropyridine (5 g, 33.8 mmol) was added to a solution of 19b (4.4 g, 33.8 mmol) and potassium tert-butoxide (7.6 g, 67.6 mmol) in tetrahydrofuran (100 ml) under ice-cooling, and the mixture was stirred for 3 hours. The insoluble material was removed by filtration, and the liquid was washed with an equal volume of water, dried over anhydrous sodium sulfate, and concentrated to dryness. The residue was purified by silica gel column chromatography to afford the desired product 20a (4.7 g, 57.0% yield).

[0261] Step 2:

[0262] Crude products 10d and 20a (134.2 mg, 0.55 mmol), anhydrous potassium carbonate (115 mg, 0.80 mmol), and Pd(dppf)Cl2 (32.5 mg, 0.04 mmol) were added sequentially to a reaction flask and dissolved in a mixed solution of 1,4-dioxane (5 mL) and water (1 mL). The atmosphere was replaced with nitrogen and stirred at 90°C for 5 h. The mixture was cooled to room temperature, the solid was filtered, and the filtrate was concentrated under reduced pressure and purified on a silica gel column to give product 20b as a yellow solid (160 mg, two-step yield 61.5%).

[0263] LCMS m / z=569.1[M+H] + .

[0264] Step 3:

[0265] At room temperature, 19d (80 mg, 0.14 mmol) was dissolved in acetonitrile (3 mL) and water (0.6 mL). Then, 1 g (CAS: 5807-14-7) (56.9 mg, 0.41 mmol) was added and the reaction continued for 24 h. The pH was adjusted to 6 with 1N hydrochloric acid, and the mixture was extracted with a 10:1 dichloromethane:methanol mixture (20 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by Pre-HPLC (instrument and preparative column: Waters 2767 Preparative HPLC, Sunfire@Prep C18, 5 μm, i.d. x length = 19 mm x 250 mm). Preparation method: The crude product was dissolved in DMF and filtered through a 0.45 μm filter to prepare the sample solution. The mobile phase system was acetonitrile / water (containing 0.5% ammonium bicarbonate). Gradient elution method: acetonitrile content 5-50% (flow rate: 15 mL / min; elution time 15 min) to obtain compound 20 (20 mg, yield: 25.8%).

[0266] LCMS m / z=555.1[M+1] + .

[0267] 1 H NMR(400MHz,DMSO-d6)δ7.88-7.80(m,6H),7.68(d,2H),7.63(d,1H),7.38(t,1H),6.93(d,1H),5.69(s,2H),5. 05-4.97(m,1H),4.63-4.56(m,1H),4.52-4.44(m,2H),4.42-4.29(m,3H),2.72-2.62(m,1H),2.37-2.28(m,1H).

[0268] Example 10: Preparation of Compound 21

[0269] first step:

[0270] 21a (7.5 g, 29.88 mmol) was added to thionyl chloride (20.0 mL) and stirred at 70°C for 2 hours. The mixture was then cooled to room temperature and concentrated to dryness. The residue was added to dichloromethane (30 mL). Ammonia (15 mL) was added to the mixture at 0°C, and a large amount of white solid precipitated. The solid was filtered and washed with water (20 mL x 2). The solid was dried to obtain 21b (7.3 g, 97% yield).

[0271] LCMS m / z=250.1[M+1] + .

[0272] Step 2:

[0273] 21b (7.3 g, 29.20 mmol) was added to phosphorus oxychloride (30.0 mL) and stirred at 100°C for 2 hours. The mixture was then cooled to room temperature and poured into ice water (100 mL). The mixture was extracted with ethyl acetate (100 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated to dryness, and the residue was purified by silica gel column chromatography to afford 21c (6.0 g, 88% yield).

[0274] LCMS m / z=232.1[M+1] + .

[0275] Step 3:

[0276] 21c (6.0 g, 25.86 mmol) was added to anhydrous ethanol (3.0 mL). The mixture was stirred thoroughly at 0°C before the addition of 33% hydrochloric acid-ethanol solution (25 mL). The temperature was gradually warmed to room temperature and stirred for 10 hours. Ether (40 mL) was then added and the mixture was filtered to obtain a solid, which was subsequently washed with ether (100 mL x 2) and dried to afford 21d (5.8 g, 71% yield).

[0277] LCMS m / z=278.1[M-Cl] + .

[0278] Step 4:

[0279] Compound 21d (3.0 g, 9.58 mmol) and compound 1d (2.5 g, 10.3 mmol) were added to glacial acetic acid (50 mL) and stirred at 60°C for 10 hours. The reaction system was cooled to room temperature and slowly added to a saturated aqueous solution of sodium bicarbonate (200 mL) for quenching. The mixture was then extracted with ethyl acetate (50 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, and filtered. The filtrate was then spin-dried to dryness, and the residue was purified by silica gel column chromatography to afford compound 21e (1.34 g, 30% yield).

[0280] LCMS m / z=457.1[M+1] + .

[0281] Step 5:

[0282] 21e (600 mg, 1.3 mmol), pinacol diboronate (519 mg, 1.9 mmol), Pd(dppf)Cl2·DCM (110 mg, 0.1 mmol), and potassium acetate (400 mg, 4.0 mmol) were added to 1,4-dioxane (10 mL). The reaction flask was replaced with nitrogen and stirred at 100°C for 5 hours. The reaction solution was cooled to room temperature, water (50 mL) was added, and extraction with ethyl acetate (50 mL x 3) was performed. The organic phases were combined, dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated to dryness to obtain crude compound 21f (438 mg, 80% yield).

[0283] LCMS m / z=423.1[M+1] + .

[0284] Step 6:

[0285] 21f (60 mg, 0.14 mmol), 3a (34 mg, 0.13 mmol), Pd(dppf)Cl2·DCM (10 mg, 0.01 mmol), and potassium carbonate (36 mg, 0.26 mmol) were added to a mixture of 1,4-dioxane (2 mL) and water (0.5 mL). The reaction flask was replaced with nitrogen and stirred at 100°C for 5 hours. The reaction solution was cooled to room temperature, water (2 mL) was added, and the mixture was extracted with ethyl acetate (10 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated to dryness, and the residue was purified by silica gel column chromatography to afford 21 g (46 mg, 54% yield).

[0286] LCMS m / z=605.2[M+1] + .

[0287] Step 7:

[0288] 21g (46mg, 0.076mmol) and 1g (32mg, 0.228mmol) were added to a mixture of acetonitrile (3mL) and water (0.5mL) and stirred at room temperature for 16 hours. The reaction solution was then adjusted to pH 6 with 1N hydrochloric acid and stirred until a white solid precipitated. The solid was filtered and washed with water. The solid was dissolved in dichloromethane, dried over anhydrous sodium sulfate, filtered, and the organic phase was concentrated to dryness. The residue was purified by silica gel column chromatography to obtain compound 21 (10mg, 22% yield).

[0289] LCMS m / z=591.0[M+1] + .

[0290] 1 H NMR(400MHz,DMSO-d6)δ12.91(br,1H),7.93-7.83(m,3H),7.78-7.67(m,3H),7.53-7.48(m,1H),7.37-7.30(m,1H),7.01-6.95(m,1 H),5.59(s,2H),5.08-4.99(m,1H),4.67-4.58(m,1H),4.55-4.45(m,2H),4.43-4.29(m,3H),2.74-2.64(m,1H),2.40-2.29(m,1H).

[0291] Example 11: Preparation of Compound 22

[0292] first step:

[0293] 21f (143 mg, 0.34 mmol), 18c (110 mg, 0.34 mmol), Pd(dppf)Cl2·DCM (40 mg, 0.04 mmol), and potassium carbonate (140 mg, 1.01 mmol) were added to a mixture of 1,4-dioxane (5 mL) and water (1 mL). The reaction flask was replaced with nitrogen and stirred at 100°C for 5 hours. The reaction solution was cooled to room temperature, water (5 mL) was added, and the mixture was extracted with ethyl acetate (10 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated to dryness, and the residue was purified by silica gel column chromatography to afford 22b (170 mg, 80% yield).

[0294] LCMS m / z=623.1[M+1] + .

[0295] Step 2:

[0296] 22b (50 mg, 0.08 mmol) and 1g (34 mg, 0.24 mmol) were added to a mixture of acetonitrile (5 mL) and water (1 mL) and stirred at room temperature for 16 hours. The reaction solution was then adjusted to pH 6 with 1N hydrochloric acid and stirred until a white solid precipitated. The solid was filtered and washed with water. The solid was dissolved in dichloromethane, dried over anhydrous sodium sulfate, filtered, and the organic phase was concentrated to dryness. The residue was separated by HPLC to afford compound 22 (10 mg, 20% yield).

[0297] LCMS m / z=609.1[M+1] + .

[0298] 1 H NMR (400MHz, DMSO-d6) δ7.95-7.90(m,1H),7.88-7.82(m,2H),7.80-7.67(m,3H),7.55-7.50(m,1H),7.37-7.31(m,1H),5.6 8(s,2H),5.06-5.00(m,1H),4.66-4.59(m,1H),4.54-4.45(m,2H),4.40-4.29(m,3H),2.69-2.64(m,1H),2.37-2.30(m,1H).

[0299] Example 12: Preparation of Compound 23

[0300] first step:

[0301] 21f (200 mg, 0.49 mmol), 20b (120 mg, 0.49 mmol), Pd(dppf)Cl2·DCM (40 mg, 0.04 mmol), and potassium carbonate (140 mg, 1.01 mmol) were added to a mixture of 1,4-dioxane (5 mL) and water (1 mL). The reaction flask was replaced with nitrogen and stirred at 100°C for 5 hours. The reaction solution was cooled to room temperature, water (5 mL) was added, and the mixture was extracted with ethyl acetate (10 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated to dryness, and the residue was purified by silica gel column chromatography to afford 23b (143 mg, 50% yield).

[0302] LCMS m / z=587.0[M+1] + .

[0303] Step 2:

[0304] 23b (50 mg, 0.08 mmol) and 1g (34 mg, 0.24 mmol) were added to a mixture of acetonitrile (5 mL) and water (1 mL) and stirred at room temperature for 16 hours. The reaction solution was then adjusted to pH 6 with 1N hydrochloric acid and stirred until a white solid precipitated. The solid was filtered and washed with water. The solid was dissolved in dichloromethane, dried over anhydrous sodium sulfate, filtered, and the organic phase was concentrated to dryness. The residue was purified by silica gel column chromatography to afford compound 23 (15 mg, 30% yield).

[0305] LCMS m / z=573.1[M+1] + .

[0306] 1H NMR(400MHz,DMSO-d6)δ7.92-7.83(m,4H),7.73-7.64(m,3H),7.53-7.47(m,1H),7.37-7.29(m,1H),7.02-6.96(m,1H),5.5 7(s,2H),5.08-5.00(m,1H),4.66-4.59(m,1H),4.54-4.45(m,2H),4.39-4.29(m,3H),2.75-2.64(m,1H),2.39-2.29(m,1H).

[0307] Formulation Example 1:

[0308] Preparation Example 1-1: Specification 5mg prescription (capsule):

[0309] Preparation Example 1-2: Specification 40mg prescription (capsule):

[0310] Preparation Example 1-3: Specification 100mg Prescription (Capsule): Y-2206004

[0311] Preparation Example 1-4: Specification 200mg prescription (capsule):

[0312] The above prescription adopts the process:

[0313] 1. Weighing: Weigh the raw and auxiliary materials according to the prescription (converted content of raw materials).

[0314] 2. Preparation of API solution: Gradually add methanol to the API until it is completely dissolved to obtain the API solution.

[0315] 3. Preparation of cyclodextrin solution: Gradually add water to cyclodextrin until it is completely dissolved to obtain cyclodextrin solution.

[0316] 4. Inclusion: After mixing the API solution and cyclodextrin solution, ultrasonicate for 1 hour to obtain a solution.

[0317] 5. Drying: Use rotary evaporation to dry the solution. After it is completely dry, take out the sample, grind and sieve it to obtain the powder.

[0318] 6. Filling: Filling is carried out according to the volume calculated after conversion of the raw material.

[0319] Formulation Example 2:

[0320] Specifications 100mg Prescription (Capsule): Y-2206003

[0321] Process:

[0322] 1. Pretreatment of API: Use air flow mill to crush the API.

[0323] 2. Weighing: Weigh the raw and auxiliary materials according to the prescription (converted content of raw materials).

[0324] 3. Mixing: The weighed tris salt of compound 6, lactose monohydrate, and sodium lauryl sulfate were mixed and sieved to obtain a mixed powder.

[0325] 4. Filling: Fill the tablets according to the weight calculated after conversion of the raw material.

[0326] Formulation Example 3:

[0327] Specifications 100mg Prescription (tablet): Y-2206005

[0328] Process:

[0329] 1. Pretreatment of API: Use air flow mill to crush the API.

[0330] 2. Weighing: Weigh the raw and auxiliary materials according to the prescription (converted content of raw materials).

[0331] 3. Premixing: Mix the weighed tris salt of compound 6, lactose monohydrate, microcrystalline cellulose, sodium lauryl sulfate, croscarmellose sodium, and colloidal silicon dioxide, and sieve the mixture to obtain a premix powder.

[0332] 4. Total mixing: Add the weighed magnesium stearate to the premixed powder and mix to obtain the total mixed powder.

[0333] 5. Tablet pressing: Tablet pressing is performed according to the calculated tablet weight after conversion of the raw material drug.

[0334] Biological test cases

[0335] 1. HEK293 / CRE-luc / GLP-1R cell activity

[0336] Cells: HEK293 / CRE-luc / GLP-1R

[0337] Cell culture medium: DMEM + 10% FBS + 400 μg / ml G418 + 100 μg / ml Hygromycin B

[0338] Freezing medium: 90% FBS, 10% (V / V) DMSO

[0339] Assay buffer: DMEM + 1% FBS

[0340] Experimental steps:

[0341] The cells were cultured in DMEM medium + 10% FBS + 400 μg / ml G418 + 100 μg / ml Hygromycin B in a 37°C CO2 incubator and passaged every 3-4 days.

[0342] Cell plating: Trypsin digestion was used to adjust the cell density to 1.67 × 10 5 cells / mL; 60 μL of cells were seeded into each well of a 384-well plate (10,000 cells / well); NC wells (negative control) and background wells (no cells) were set up. Incubate in an incubator for approximately 18 ± 2 h.

[0343] The compound was diluted with detection buffer in a gradient manner, with the detection concentration ranging from 0.01 nM to 1000 nM.

[0344] Remove the cell culture plate, aspirate all supernatant from the cells, and gently wash twice with 1X PBS.

[0345] Add the diluted compound to a 384-well plate (10 μL / well), with three replicates for each concentration. Add 10 μL of detection buffer to the NC wells, seal, and incubate at 37°C for 6 h.

[0346] The plate was removed and the cells were allowed to equilibrate to room temperature (at least 15 min), after which all the supernatant was aspirated from the cells.

[0347] Add 10 μL / well to the sample wells Reagent and incubate at room temperature for 5 min to lyse the cells.

[0348] The test results were read using a BMG microplate reader.

[0349] Data processing:

[0350] Calculate the average background value.

[0351] The fold of induction (FI) was calculated as (value of the induced wells - background value) / (value of the negative control wells - background value).

[0352] Graphpad Prim 8.0 software was used to calculate the EC of the samples using a four-parameter fitting analysis. 50 Numeric value.

[0353] Statistical analysis

[0354] All results are presented as mean ± SEM, and statistical analysis was performed using Graphpad Prism software. Specific data are presented in graphical form. P < 0.05 was considered statistically significant.

[0355] Biological test results:

[0356] A<10nM

[0357] Conclusion: The compounds of the present invention, such as the compounds in the examples, have a good agonist effect on the GLP-1 receptor. 50 The value is less than 100 nM, for example, the EC values ​​of compound 6, compound 7, compound 8, compound 9, compound 10, compound 11, compound 18, compound 20, compound 21, compound 22, and compound 23 50 The value was less than 10 nM.

[0358] 2. Effects of compounds on hERG potassium channels

[0359] Experimental platform: electrophysiology manual patch clamp system

[0360] Cell line: Chinese hamster ovary (CHO) cells stably expressing hERG potassium channel

[0361] Experimental method: CHO (Chinese Hamster Ovary) cells stably expressing hERG potassium channel were used to record hERG potassium channel current using whole-cell patch clamp technique at room temperature. Glass microelectrodes were made from glass electrode blanks (BF150-86-10, Sutter) using a puller. The tip resistance after perfusing the electrode liquid was about 2-5MΩ. The glass microelectrode was inserted into the amplifier probe to connect to the patch clamp amplifier. The clamping voltage and data recording were controlled and recorded by computer using pClamp 10 software, with a sampling frequency of 10kHz and a filter frequency of 2kHz. After obtaining the whole-cell recording, the cell was clamped at -80mV to induce hERG potassium current (I hERG ) was administered with a 2-second depolarization step from -80 mV to +20 mV, followed by repolarization to -50 mV for 1 second before returning to -80 mV. This voltage stimulus was administered every 10 seconds, and administration began after confirming that the hERG potassium current was stable (for at least 1 minute). Compound administration was continued for at least 1 minute at each test concentration, and at least two cells (n ≥ 2) were tested for each concentration.

[0362] Data processing:

[0363] Data were analyzed and processed using pClamp 10, GraphPad Prism 5, and Excel software. The degree of inhibition of hERG potassium current (peak hERG tail current evoked at -50 mV) by different compound concentrations was calculated using the following formula: Inhibition% = [1–(I / Io)] × 100%.

[0364] Wherein, Inhibition% represents the inhibition percentage of the compound on hERG potassium current, and I and Io represent the amplitude of hERG potassium current before and after drug addition, respectively.

[0365] Compound IC 50 The results were calculated using GraphPad Prism 5 software by fitting the following equation: Y = Bottom + (Top-Bottom) / (1 + 10^((LogIC 50 -X)×HillSlope))

[0366] Where X is the Log value of the test sample concentration, Y is the inhibition percentage at the corresponding concentration, and Bottom and Top are the minimum and maximum inhibition percentages, respectively.

[0367] Biological test results

[0368] Conclusion: The compounds of the present invention, such as the compounds in the Examples, have a smaller hERG inhibitory effect.

[0369] 3. Mouse Liver Microsome Stability Test

[0370] 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) ).

[0371] Biological test results:

[0372] Conclusion: The compounds of the present invention, such as the example compounds, have good stability in mouse liver microsomes. For example, the clearance rate of compound 6 in mouse liver microsomes was significantly slowed down and the half-life was significantly prolonged compared with the control compound. 1 / 2 The ratio is greater than 2.

[0373] 4. Human Liver Microsome Stability Test

[0374] At 37°C, 1 μM of the test compound was incubated with human liver microsomes (0.5 mg / mL) supplemented with 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 human 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) ).

[0375] Biological test results:

[0376] Conclusion: The compounds of the present invention, such as the example compounds, have good stability in human liver microsomes. For example, the clearance rate of compound 6 in human liver microsomes is significantly slowed down and the half-life is significantly prolonged compared with the control compound. 1 / 2 The ratio is greater than 3.

[0377] 5. Pharmacokinetic testing

[0378] Pharmacokinetic evaluation in mice

[0379] Male C57 mice (purchased from Hunan Slake Jingda Experimental Animal Co., Ltd., license number: SCXK (Xiang) 2019-0004), 20-25g, fasted overnight. On the day of the experiment, 18 male C57 mice were divided into two groups, 9 in each group, and the test compound was administered intravenously and orally at doses of 5 mg / kg and 20 mg / kg, respectively. 0.1 ml of blood was collected from the orbital venous plexus before administration and 5, 15, 30 minutes, 1, 2, 4, 6, 8 and 24 hours after administration. The blood was placed in an EDTAK2 centrifuge tube and centrifuged at 6000 rpm and 4 ° C for 5 minutes to collect plasma. The plasma sample was added to an acetonitrile solution containing an internal standard (dexamethasone, 500 ng / mL), vortexed and centrifuged at 10000 rpm for 10 minutes, and the supernatant was taken for LC-MS / MS analysis. Pharmacokinetic parameters were calculated using the non-compartmental model in WinNonlin Version 8.0 (Pharsight, Mountain View, CA) pharmacokinetic software.

[0380] Conclusion: The compounds of the present invention, such as the compounds in the examples, have good oral properties. For example, the bioavailability of compound 6 in C57 mice is >25%.

[0381] Pharmacokinetic evaluation in cynomolgus monkeys

[0382] Male cynomolgus macaques (housed at Suzhou Leo Biotechnology Co., Ltd.) weighing 3-5 kg ​​were fasted overnight (14-18 hours). On the day of the experiment, six male cynomolgus macaques were divided into two groups of three and administered the test compound at doses of 2 mg / kg and 10 mg / kg intravenously and orally. Blood (1 ml) was collected from the four limb veins before administration and at 5, 15, 30 minutes, 1, 2, 4, 6, 8, 10, 12, and 24 hours after administration. The blood was placed in EDTAK2 centrifuge tubes and centrifuged at 6000 rpm at 4°C for 5 minutes to collect plasma. Plasma samples were spiked with acetonitrile containing an internal standard (dexamethasone, 500 ng / mL), vortexed, and centrifuged at 10,000 rpm for 10 minutes. The supernatant was then analyzed by LC-MS / MS. Pharmacokinetic parameters were calculated using a non-compartmental model in WinNonlin Version 8.0 (Pharsight, Mountain View, CA) pharmacokinetic software.

[0383] Conclusion: The compounds of the present invention, such as the compounds in the examples, have good metabolic properties. For example, compound 6 has a low clearance rate and a long half-life in monkeys, which has the advantages of longer drug action and less frequent dosing.

[0384] 6. Cytochrome P450 isoenzyme inhibition test

[0385] Experimental purpose: To determine the inhibitory effect of the test compounds on the activities of human liver microsomal cytochrome P450 isoenzymes (CYP1A2, CYP2C19, CYP2D6 and CYP3A4).

[0386] Experimental procedure: The test compounds at incubation concentrations of 0, 0.03, 0.1, 0.3, 1, 3, 10, 30 and 100 μM were pre-incubated with human liver microsome solutions containing 5 enzyme mixed substrates (liver microsome concentration: 0.1 mg / mL) in a 37°C water bath for 10 minutes, and then the coenzyme factor (NADPH) was added to initiate the reaction; after further incubation in a 37°C water bath for 10 minutes, the reaction was terminated by adding glacial acetonitrile solution containing the internal standard, shaken for 1 minute, and centrifuged at 4°C and 10,000 rpm for 10 minutes. The supernatant was then subjected to LC / MS / MS analysis to detect the amount of metabolites produced by each CYP enzyme substrate.

[0387] The production rate of each metabolite reflects the activity of each CYP isoenzyme. The activity of each isoenzyme in the solvent control incubation system without the test substance was set to 100%, and the ratio of the production rate of the metabolite in the presence of different concentrations of the test substance to the production rate of the metabolite in the solvent control sample was used as the residual activity percentage of each isoenzyme. The data were plotted using Graphpad Prism 5.0 with the residual activity percentage as the ordinate and the inhibitor concentration as the abscissa, and the IC value of the test substance for each CYP isoenzyme was calculated.50 value.

[0388] Conclusion: The compounds of the present invention, such as the example compounds, have weak inhibitory activity against cytochrome P450 isoenzymes. For example, compound 6 has inhibitory activity IC of CYP2C19, CYP1A2, CYP2D6 and CYP3A4-M. 50 >50μM.

[0389] 7. Intravenous Glucose Tolerance Test in Cynomolgus Monkeys

[0390] Experimental animals: 6 male cynomolgus macaques (raised at Crown Biotech (Taicang) Co., Ltd.), weighing 6.5-7.5 kg.

[0391] Experimental procedure: During the entire experimental period, before each blood collection (ivGTT operation), the experimental animals were required to fast overnight for approximately 16 hours. The animals were grouped and dosed according to the following Table 10-1.

[0392] Intravenous Glucose Tolerance Test (ivGTT): After anesthesia, baseline blood was collected, followed by an IV bolus and infusion. Pre-glucose baseline blood glucose was collected 15 minutes after the infusion, and 50% glucose solution (0.25 g / kg, 0.5 mL / kg) was administered 30 minutes after the infusion. The total infusion duration was 1.0 hour.

[0393] Blood was collected before administration, at -15 minutes (15 minutes before glucose administration), at 0 minutes (before glucose administration), and at 2, 5, 7, 10, 20, and 30 minutes after glucose administration. The collected blood was allowed to stand at room temperature for at least 30 minutes and then centrifuged at 3500 g / min for 10 minutes at room temperature. The supernatant was aliquoted and stored at 2-8°C until glucose and insulin analysis. All experimental data were analyzed using GraphPad Prism 9.3.1.

[0394] Table 10-1 Grouping and Dosage

[0395] Note: *Intravenous infusion was performed at a constant rate using a Harvard syringe pump. The solvent consisted of 5% polyethylene glycol 400 (PEG400) + 95% [20% sulfobutyl-β-cyclodextrin (SEBCD)]. After brief sonication, the solvent was completely dissolved and filtered through a disposable syringe filter before preparation. Prepare the solution immediately and adjust the pH to 7.0-7.4.

[0396] The test results are shown in Table 10-2:

[0397] Table 10-2 Blood glucose and insulin analysis results

[0398] Conclusion: The compounds of the present invention, such as the compounds in the examples, can promote insulin secretion, improve glucose tolerance and increase glucose clearance rate.

[0399] 8.Caco2 permeability test

[0400] The assay 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 a compound of the invention (2 μM) or control compounds: Compound DZ-6, 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 by LC-MS / MS, and the concentrations of the compound of the invention and control compound were determined. Concentration data were used to calculate the apparent permeability coefficients for transport from the apical to basolateral and basolateral to apical sides of the cell monolayer, thereby calculating the efflux rate. Leakage of Lucifer Yellow was used to assess the integrity of the cell monolayer after 2 hours of incubation.

[0401] Table 11-1. Caco2 test results of the compounds of the present invention

[0402] Conclusion: The compounds of the present invention, such as the example compounds, are low permeability compounds. For example, the efflux rate of compound 6 is lower than that of the control compound DZ-6.

[0403] 9. Pharmacokinetic testing of preparations in monkeys

[0404] Experimental animals: Male cynomolgus monkeys, 3-5 kg, 3-5 years old, 3 per group, purchased from Suzhou Xishan Zhongke Experimental Animal Co., Ltd.

[0405] Experimental method: On the day of the experiment, monkeys were randomly divided into groups according to body weight, 3 per group. They were fasted but not watered for 14-18 hours before administration and fed 4 hours after administration.

[0406] Table 12-1. Dosage Information

[0407] *The dosage is based on Tris salt. 100 mg of Tris salt contains 82.54 mg of free Tris salt.

[0408] Before and after dosing, 1.0 mL of blood was collected from a limb vein and placed in an EDTAK2 centrifuge tube. The tubes were centrifuged at 5000 rpm at 4°C for 10 minutes to collect plasma. For the oral gavage group, samples were collected at 0, 5, 15, 30 minutes, and 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.

[0409] Table 12-2. Pharmacokinetic parameters of test samples in monkey plasma

[0410] 10. Oral Glucose Tolerance Test (OGTT) in Mice

[0411] An OGTT experiment was performed in male C57BL / 6-G1p1rem2 (hGLP1R) Smoc mice to evaluate the hypoglycemic effect of GLP-1 receptor agonists. The animals were fasted for more than 16 hours and randomly divided into groups for tail tip blood sampling. Basal fasting blood glucose (FBG) was measured using a blood glucose meter. Mice with FBG within the range of 3.5 to 6.5 mmol / L were orally administered (the dosage was 3.63 mg / kg in terms of tris salt), and the control group was given the vehicle. 15 minutes after administration, each group of animals was gavaged with 25% glucose solution. Blood glucose levels were measured at 15 minutes, 30 minutes, 45 minutes, 1 hour, and 2 hours after administration, and 0-2 hour blood glucose AUC was calculated using Graphpad Prism software. 0-2h The inhibition rate was calculated.

[0412] Calculation formula: Blood glucose inhibition rate = (AUC 0-2h -AUC of the drug-treated group 0-2h ) / AUC of vehicle group 0-2h ×100%

[0413] Results: Compound 6 at 3.63 mg / kg (as tris salt, po, single) in the OGTT test blood glucose AUC in hGLP1R mice 0-2h A decrease of 43.19%.

Claims

1. A pharmaceutical composition or pharmaceutical preparation, wherein the pharmaceutical composition or pharmaceutical preparation comprises an active ingredient A and a pharmaceutical excipient, wherein the active ingredient A is selected from the compound of general formula (I) or its stereoisomers, tautomers, deuterated substances, solvates, prodrugs, metabolites, pharmaceutically acceptable salts or cocrystals, Ring B is selected from C 6-10 Aromatic ring or 5 to 12 membered heteroaromatic ring, said aromatic ring or heteroaromatic ring is optionally substituted by 1 to 4 R B Substitution, the heterocyclic group or heteroaromatic ring contains 1 to 3 heteroatoms selected from O, S and N; Ring C is selected from C 6-10 Carbocyclic, 5- to 10-membered heterocyclic, C 6-10 aromatic ring or 5 to 10 membered heteroaromatic ring, wherein the carbocyclic ring, heterocyclic ring, aromatic ring or heteroaromatic ring is optionally substituted by 1 to 4 R C Substituted, the heterocyclic or heteroaromatic ring contains 1 to 5 heteroatoms selected from O, S and N; L is selected from a bond, O, S, -NR L -, -C(R L )2-, -C(R L )2-C(R L )2-, -Y-C(R L )2-, -C(R L )2-Y-, -Y-C(R L )2-C(R L )2-, -C(R L )2-C(R L )2-Y- or -C(R L )2-Y-C(R L )2-; Y is selected from O, S or -NR L -; R1 is selected from H, halogen, OH, -SH, CF3, CN, C 1-6 Alkyl, C 1-6 Alkoxy, -C 1-3 Alkylene-ZC 0-3 Alkylene-R 1a 、C 0-4 Alkylene-R 1a The alkyl, alkoxy, and alkylene groups are optionally substituted by 1 to 4 groups selected from halogen, =O, CN, OH, -N(R 1b )2、C 1-6 Alkyl, halogen-substituted C 1-6 Alkyl, hydroxy substituted C 1-6 Alkyl, cyano substituted C 1-6 Alkyl, C 1-6 Alkoxy, C 3-6 substituted by a cycloalkyl, a 3- to 6-membered heterocycloalkyl, or a 5- to 10-membered heteroaryl substituent, wherein the heterocycloalkyl or heteroaryl group contains 1 to 3 heteroatoms selected from O, S, and N; Z is selected from a bond, N(R 1b ), O or S; R 1a Selected from C 1-6 Alkyl, C 1-6 Alkoxy, C 3-8 Cycloalkyl, 3 to 8 membered heterocycloalkyl, 6 to 10 membered aryl or 5 to 12 membered heteroaryl, wherein the cycloalkyl, heterocycloalkyl, aryl, heteroaryl is optionally substituted by 1 to 4 groups selected from halogen, =O, OH, CN, C 1-6 Alkyl, C 1-6 Alkoxy or -N(R 1b )2 is substituted by a substituent, wherein the alkyl group or alkoxy group is optionally substituted by 1 to 4 substituents selected from H, halogen, =O, CN, OH, -N(R 1b )2、C 1-6 Alkyl, C 1-6 Alkoxy, C 3-6 Substituted by a cycloalkyl substituent, wherein the heterocycloalkyl or heteroaryl group contains 1 to 3 heteroatoms selected from O, S, and N; R 1b Each independently selected from H or C 1-6 Alkyl, said alkyl being optionally substituted by 1 to 3 groups selected from halogen, =O, CN, OH, NH2, C 1-6 Alkyl, C 1-6 Alkoxy or C 3-6 substituted by a cycloalkyl substituent; R L Each independently selected from H, halogen, -SH, CN, OH, CF3, NH2, C 1-6 Alkyl, C 1-6 Alkoxy or C 3-6 Cycloalkyl, wherein the alkyl, cycloalkyl or alkoxy is optionally substituted by 1 to 4 groups selected from halogen, =O, OH, CN, NH2, C 1-6 Alkyl, C 3-6 Cycloalkyl or C 1-6 substituted by an alkoxy substituent; R B 、R C or R D Each independently selected from H, halogen, =O, CN, CF3, OH, -SH, NH2, C 1-6 Alkyl, C 3-6 Cycloalkyl or C 1-6 Alkoxy, wherein the alkyl, cycloalkyl or alkoxy is optionally substituted by 1 to 4 groups selected from halogen, =O, OH, CN, NH2, C 1-6 Alkyl, C 3-6 Cycloalkyl or C 1-6 substituted by an alkoxy substituent; Alternatively, R C With R L Each of the atoms and the atoms connected to it together form C 3-10 Carbocyclic or 3 to 10 membered heterocyclic ring, said carbocyclic or heterocyclic ring being optionally substituted by 1 to 4 H, halogen, =O, OH, CN, NH2, C 1-6 Alkyl, C 3-6 Cycloalkyl or C 1-6 Alkoxy is substituted by a substituent, wherein the alkyl, cycloalkyl, alkoxy is optionally substituted by 1 to 4 groups selected from halogen, =O, OH, CN, NH2, C 1-6 Alkyl, C 3-6 Cycloalkyl or C 1-6 substituted by an alkoxy substituent, wherein the heterocyclic ring contains 1 to 3 heteroatoms selected from O, S and N; m is selected from 0, 1, 2, 3 or 4; The pharmaceutical composition or pharmaceutical preparation contains 1-600 mg of active ingredient A.

2. The pharmaceutical composition or pharmaceutical preparation according to claim 1, wherein the pharmaceutical composition or pharmaceutical preparation comprises 5-300 mg of active ingredient A.

3. The pharmaceutical composition or pharmaceutical preparation according to claim 2, wherein the pharmaceutical composition or pharmaceutical preparation comprises 5-200 mg of active ingredient A.

4. The pharmaceutical composition or pharmaceutical preparation according to claim 3, wherein the pharmaceutical composition or pharmaceutical preparation comprises 5-100 mg of active ingredient A.

5. The pharmaceutical composition or pharmaceutical formulation according to claim 4, wherein the pharmaceutical composition or pharmaceutical formulation comprises 5 mg, 10 mg, 20 mg, 40 mg, 100 mg or 200 mg of active ingredient A.

6. The pharmaceutical composition or pharmaceutical preparation according to claim 1, wherein the pharmaceutical composition or pharmaceutical preparation comprises an active ingredient A and a pharmaceutical excipient, wherein the active ingredient A is selected from the compound of formula (I) or its stereoisomers, tautomers, deuterated substances, solvates, prodrugs, metabolites, pharmaceutically acceptable salts or cocrystals, Ring B is selected from one of the following substituted or unsubstituted groups: Its left side is directly connected to ring C and, when substituted, is optionally substituted by 1, 2 or 3 R B replace; Ring C is selected from one of the following substituted or unsubstituted groups: When substituted, optionally 1, 2, or 3 Rs C Replace, its left side is connected to L; L is selected from -CH2O-, -OCH2-, -CH2NH-, -NHCH2-, -CH2CH2-, -OCH2CH2-, -CH2CH2O-, -NHCH2CH2-, -CH2CH2NH-, and the right side is connected to ring C; or Selected from Its right side is connected to ring B, wherein the benzene ring is optionally substituted by 1, 2 or 3 R C replace; R1 is selected from R B 、R C or R D Each is independently selected from H, F, Cl, CN, CF3, CHF2, methyl, ethyl, methoxy, ethoxy, tert-butyloxy, and trifluoromethoxy.

7. The pharmaceutical composition or pharmaceutical formulation according to claim 6, wherein the pharmaceutical composition or pharmaceutical formulation comprises an active ingredient A and a pharmaceutical excipient, wherein the active ingredient A is selected from the compound of general formula (I) or its stereoisomers, tautomers, deuterated forms, solvates, prodrugs, metabolites, pharmaceutically acceptable salts or cocrystals, wherein the structure of the compound is selected from one of the structures shown in Table S-1.

8. The pharmaceutical composition or pharmaceutical preparation according to claim 1, wherein the pharmaceutically acceptable salt of the compound of formula (I) is selected from tris(hydroxymethylaminomethane) salt or triethanolamine salt.

9. The pharmaceutical composition or pharmaceutical preparation according to claim 1, wherein the active ingredient A is selected from a tris(hydroxymethyl)aminomethane salt having the following structure:

10. The pharmaceutical composition or pharmaceutical preparation according to claim 1, wherein the active ingredient A is selected from the triethanolamine salt of the following structure:

11. A pharmaceutical composition or pharmaceutical preparation comprising the active ingredient A according to any one of claims 1 to 10 and a pharmaceutically acceptable excipient, wherein the pharmaceutically acceptable excipient comprises hydroxypropyl-β-cyclodextrin.

12. The pharmaceutical composition or pharmaceutical preparation according to claim 11, wherein the content of active ingredient A is 0.5%-15.4%.

13. A pharmaceutical composition or pharmaceutical preparation comprising the active ingredient A according to claim 1 and a pharmaceutical excipient, wherein the pharmaceutical excipient comprises lactose (preferably lactose monohydrate) or sodium lauryl sulfate, and preferably, the content of the active ingredient A is 20%-25%.

14. The pharmaceutical composition or pharmaceutical preparation according to claim 13, wherein the pharmaceutical excipient further comprises one or more of microcrystalline cellulose, croscarmellose sodium, silicon dioxide or magnesium stearate.

15. A method for treating a disease in a mammal, said method comprising administering to a subject a therapeutically effective amount of active ingredient A, preferably 1-600 mg, said disease being preferably diabetes.

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