Novel oxopyridine compounds and preparation methods and uses thereof

Abstract

The present invention discloses a compound represented by formula (I), a stereoisomer thereof, or a pharmaceutically acceptable salt thereof. The present invention also provides the use of the compound, its stereoisomer or pharmaceutically acceptable salt in the preparation of a drug for treating and / or preventing diseases associated with FXIa receptors, especially in the preparation of a drug for treating and / or preventing cerebrovascular arterial disease and / or peripheral arterial disease.

Description

Technical Field

[0001] The present invention relates to the field of pharmaceutical chemistry, and in particular to oxopyridine compounds or their salts, isomers, preparation methods thereof, and use thereof in preparing drugs for treating and / or preventing diseases associated with FXIa receptors, especially in preparing drugs for treating and / or preventing cerebrovascular arterial diseases and / or peripheral arterial diseases. Background Art

[0002] Thromboembolic disease is a disease caused by abnormal blood clots formed in blood vessels during the survival of humans and animals. There are three reasons for thrombosis: damaged blood vessels, blood changes and blood stasis; it is a group of complications caused by many different diseases and different reasons. Due to the differences in various underlying diseases and the different sites of thromboembolism, thrombosis may clinically manifest as myocardial infarction, stroke, deep vein thrombosis (DVT), pulmonary embolism, atrial fibrillation and cerebral infarction, etc., especially myocardial infarction, cerebral infarction and pulmonary infarction with embolism and infarction as the main causes, which ranks first among various causes of death, claiming nearly 12 million lives each year worldwide, close to a quarter of the world's total death toll.

[0003] Coagulation factor XI (FXI) is a plasma serine protease necessary for maintaining the intrinsic pathway. After activation, it generates activated coagulation factor XIa (FXIa), which plays a key role in the amplification of the coagulation cascade. In the coagulation cascade, thrombin can feedback activate FXI, and activated FXI in turn promotes the massive production of thrombin, thereby amplifying the coagulation cascade. Therefore, drugs targeting the FXI target can block the intrinsic pathway and inhibit the amplification of the coagulation cascade, thus having an anti-thrombotic effect. In recent years, clinical data on the association between human coagulation factor XI (FXI) deficiency or elevated FXI levels and the occurrence of thrombotic diseases, as well as antithrombotic experimental studies on animals with FXI deficiency, knockout or inhibition have shown that compared with direct FXa inhibitors, inhibition of FXI may have a lower risk of bleeding and is a new target for antithrombotic prevention and treatment.

[0004] The reported FXI inhibitors mainly include monoclonal antibodies, antisense oligonucleotides, chemical small molecules, peptides or proteins and peptide mimetics. At present, milvexian, jointly developed by BMS and Johnson & Johnson, has completed the Phase II clinical trial, and the results show that it has a lower risk of bleeding. The Phase I clinical trial of BMS's intravenous small molecule FXIa inhibitor BMS-962122 has been completed and research and development has been suspended. ONO-7684, a small molecule oral FXIa inhibitor developed by Ono Co., Ltd. of Japan, has entered the Phase I clinical study. Monoclonal antibodies and antisense oligonucleotides need to be injected, and they have the disadvantages of being expensive, slow to take effect and possibly difficult to control. Chemical small molecules have the advantages of relatively good oral bioavailability and better patient compliance. Therefore, the development of safe, effective, specific and active new FXIa small molecule inhibitors may make up for the shortcomings of current clinical anticoagulant and antithrombotic drugs that are prone to bleeding complications and meet unmet clinical needs. Summary of the invention

[0005] The present invention provides a novel oxopyridine compound. Most of the compounds in the embodiments show good anticoagulant effect in animals and in vitro affinity for FXIa.

[0006] In one aspect, the present invention provides a compound represented by formula (I), a stereoisomer or a pharmaceutically acceptable salt thereof:

[0007]

[0008] in,

[0009] X is selected from C or N;

[0010] R 1 Selected from (CH2) f COOR 5 or (CH2) f CONR 6 R 7 , where: R 5 is selected from H, alkyl; R 6 , R 7 are independently selected from hydrogen or alkyl, or R 6 , R 7 The ring is closed to form a substituted or unsubstituted five-membered heterocyclic ring or a six-membered heterocyclic ring, and the substituent is selected from a substituted or unsubstituted alkyl group, a substituted or unsubstituted cycloalkyl group, or a substituted or unsubstituted aryl group; f is selected from an integer from 0 to 3;

[0011] R 2 Independently selected from COOR 8 or CONR 9 R 10 , where R 8 , R 9 , R10 are independently selected from hydrogen, alkyl or cycloalkyl;

[0012] R 3 are independently selected from hydrogen, halogen, alkoxy or haloalkyl;

[0013] R 4 Independently selected from tetrazole, or substituted or unsubstituted triazole, the substituent of the triazole is selected from halogen or halogenated alkyl.

[0014] Furthermore, the above f is selected from 0, 1, 2 or 3; preferably 0 or 1.

[0015] Furthermore, in the above compound, its stereoisomer or pharmaceutically acceptable salt, the R 1 Selected from (CH2) f COOR 5 or (CH2) f CONR 6 R 7 , where R 5 Selected from H or C1~C6 alkyl;

[0016] R 6 , R 7 The ring is closed to form a substituted or unsubstituted five-membered heterocyclic ring or six-membered heterocyclic ring, wherein the heterocyclic ring includes:

[0017] ,

[0018] Among them, R 11 is selected from alkyl, cycloalkyl, hydroxyalkyl, or substituted or unsubstituted aryl, wherein the substituent of the aryl is selected from halogen.

[0019] Further, in the above compound, its stereoisomer or pharmaceutically acceptable salt, X is selected from C or N;

[0020] or / and R 2 Independently selected from COOR 8 or CONR 9 R 10 , where: R 8 , R 9 , R 10 Independently selected from hydrogen, C1~C6 alkyl or C3~C6 cycloalkyl;

[0021] or / and R 3 Selected from hydrogen, fluorine, chlorine, bromine, trifluoromethyl or C1~C4 alkoxy;

[0022] or / and R 4 Selected from tetrazole, or substituted or unsubstituted triazole, wherein the substituent of the triazole is selected from fluorine, chlorine, bromine or trifluoromethyl;

[0023] or / and R 5 Selected from C1~C4 alkyl;

[0024] or / and R 11 is selected from C1~C6 alkyl, C3~C6 cycloalkyl, hydroxy-substituted C1~C6 alkyl, or substituted or unsubstituted phenyl, wherein the substituent of the phenyl is selected from halogen;

[0025] or / and R 10 is selected from substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C1-C6 cycloalkyl, substituted or unsubstituted phenyl, or substituted or unsubstituted pyridyl.

[0026] Further, in the above compound, its stereoisomer or pharmaceutically acceptable salt, X is selected from C or N;

[0027] or / and R 2 Independently selected from COOR 8 ,CONR 9 R 10 , where: R 8 , R 9 , R 10 independently selected from hydrogen, methyl, ethyl, propyl, tert-butyl or cyclopropyl;

[0028] or / and R 3 is selected from hydrogen, fluorine, chlorine, bromine, trifluoromethyl or methoxy;

[0029] or / and R 4 is selected from substituted or unsubstituted triazoles, wherein the substituent of the triazole is selected from chlorine or trifluoromethyl;

[0030] or / and R 5 is selected from methyl, ethyl, propyl, tert-butyl or cyclopropyl;

[0031] or / and R 11 Selected from methyl, ethyl, propyl, butyl, hydroxyethyl, cyclopropyl, cyclopropylmethyl, phenyl, 4-fluorophenyl, 2-fluorophenyl or 2,4-difluorophenyl.

[0032] Furthermore, the above-mentioned compound, its stereoisomer or pharmaceutically acceptable salt includes the following compounds:

[0033]

[0034]

[0035]

[0036]

[0037]

[0038]

[0039]

[0040]

[0041] In case of conflict between chemical nomenclature and structural formula in the present invention, the compound structure represented by the number shall prevail.

[0042] Furthermore, in the compound of formula (1), its stereoisomer or pharmaceutically acceptable salt, the hydrogen in the compound, its stereoisomer or pharmaceutically acceptable salt may be replaced by one or more deuterium.

[0043] In another aspect, the present invention provides a method for preparing the compound of formula (1), its stereoisomers or pharmaceutically acceptable salts, comprising the following steps:

[0044] When R 1 For (CH2) f COOR 5 , R 5 When selected from alkyl, the synthetic route comprises the following steps:

[0045]

[0046] Step 1: The starting material a undergoes bromination reaction to generate intermediate b;

[0047] Step 2: Intermediate b undergoes hydrolysis reaction to generate intermediate c;

[0048] Step 3: Intermediate c undergoes substitution reaction with compound d to generate intermediate e;

[0049] Step 4: Intermediate e undergoes hydrolysis reaction to generate compound f;

[0050] Step 5: The intermediate f undergoes a condensation reaction with the compound g to generate a compound of formula (1);

[0051] When R 1 For (CH2) f When COOH, the synthesis route includes the following steps:

[0052]

[0053] Compound h undergoes a hydrolysis reaction to generate a compound of formula (1);

[0054] When R 1 For (CH2)f CONR 6 R 7 , R 2 For COOR 8 or CONR 9 R 10 , R 8 When selected from alkyl or cycloalkyl, the synthetic route comprises the following steps:

[0055]

[0056] Compound i undergoes a condensation reaction with the corresponding amine to generate a compound of formula (1);

[0057] When R 1 For (CH2) f CONR 6 R 7 , R 2 When it is COOH, the synthesis route comprises the following steps:

[0058]

[0059] Compound j undergoes a hydrolysis reaction to generate a compound of formula (1);

[0060] Wherein, unless otherwise specified, the above X, R 1 ~R 10 , f are defined as any of the above corresponding definitions.

[0061] In a third aspect, the present invention also provides the use of the above-mentioned compound, its stereoisomer or pharmaceutically acceptable salt in the preparation of a drug for treating and / or preventing diseases associated with FXIa receptor.

[0062] Furthermore, the above-mentioned disease associated with FXIa receptor is selected from cerebrovascular arterial disease and / or peripheral arterial disease.

[0063] Furthermore, the above-mentioned diseases associated with FXIa receptors are selected from transient ischemic attack (TIA), ischemic stroke, including cardiogenic stroke, such as stroke caused by atrial fibrillation, non-cardiogenic stroke, such as lacunar stroke, stroke caused by large artery or small artery disease, or stroke caused by undetermined causes, cryptogenic stroke, embolic stroke, embolic stroke of undetermined origin, or events of thrombotic and / or thromboembolic origin leading to stroke or TIA, and / or peripheral arterial disorders leading to peripheral arterial disease, including peripheral arterial occlusion, acute limb ischemia, amputation, reocclusion and restenosis after intervention (such as angioplasty, stent implantation or surgery and bypass), and / or stent thrombosis.

[0064] The term "alkyl" of the present invention refers to a C1-C12 lower straight chain and a straight chain saturated alkyl group.

[0065] Beneficial effects: Compared with the prior art, the compounds of the present invention have better in vitro anticoagulant effects, and are highly consistent with the inhibitory effects on FXIa tested in vitro. At the same time, in some embodiments, the compounds of the present invention have better rat injection pharmacokinetic characteristics, such as better AUC, and have the characteristics of rapid elimination. While achieving drug efficacy, the potential bleeding risk caused by long-term action is reduced, and the safety is higher. DETAILED DESCRIPTION

[0066] The present invention will be further described in detail below in conjunction with embodiments and test examples. The embodiments and test examples of the present invention are only used to illustrate the technical scheme of the present invention, and are not intended to limit the present invention. Any equivalent substitutions in the art made in accordance with the contents disclosed in the present invention shall fall within the protection scope of the present invention.

[0067] The compounds of the present invention, their stereoisomers or pharmaceutically acceptable salts can be prepared by selecting the synthetic routes of the embodiments, and the conventional conditions of the reaction raw materials and the reaction solvent can be adjusted according to the needs of the substituents or the salt formation, which can be realized by those skilled in the art based on the disclosure of the present invention. In addition, the column chromatography of the present invention refers to silica gel column chromatography unless otherwise specified, and the elution solvent can be determined as a single or mixed elution solvent in combination with the reaction solvent and the common knowledge or common means of those skilled in the art unless otherwise specified.

[0068] The structures of the compounds were determined by NMR ( 1 H NMR) or liquid chromatography-mass spectrometry (LC-MS).

[0069] The liquid chromatography-mass spectrometer (LC-MS) was Agilent G6120B (used with Agilent 1260 liquid phase); the nuclear magnetic resonance instrument ( 1 HNMR) was Bruker AVANCE-400 or Bruker AVANCE-800, NMR ( 1 H NMR) shift ( <h2 style=";text-align:left;direction:ltr">δ ) are given in parts per million (ppm), the solvent used was DMSO, the internal standard was tetramethylsilane (TMS), and the chemical shift was based on 10 -6 (ppm) is given as the unit.

[0070] The term "room temperature" in the present invention means a temperature between 10 and 30°C.

[0071] Example 1: Preparation of Compound 1

[0072]

[0073] Step 1: Preparation of intermediate b

[0074] Take 8.5g (86.73mmol) of concentrated sulfuric acid, add it to 50ml of water, cool it to below 5℃, add 14.8g (124.4mmol) of potassium bromide, 5g (31.03mmol) of D-glutamic acid-5-methyl ester, and drop 3.2g (46.38mmol) of sodium nitrite dissolved in 25ml of water, control the temperature below 0~5℃, and add it in about 30 minutes. After the addition, stir and react at 0~5℃ overnight. The next day, the reaction solution was extracted three times with ethyl acetate, the organic phases were combined, dried over anhydrous sodium sulfate, and the solvent was evaporated to obtain 8g of crude product. Purify it through a column, eluent (ethyl acetate: petroleum ether = 1:1), collect the product to obtain 2.9g of oil. The yield is 41.5%, and the purity is 95.91%.

[0075] ESI-MS: m / z = 225.1(M+H) + .

[0076] Step 2: Preparation of intermediate c

[0077] At room temperature, take 2.9g of intermediate b, add 30ml of dichloromethane, 3.9g (38.54mmol) of triethylamine, 0.31g (2.54mmol) of DMAP, and dropwise add 4.2g (19.24mmol) of di-tert-butyl dicarbonate dissolved in 30ml of dichloromethane, and add it for about 30 minutes. After the addition, react at room temperature for 30 minutes. After the reaction is completed, add water and dichloromethane to extract, and the organic phase is washed with 5% citric acid, saturated sodium bicarbonate, water, saturated salt water, dried with anhydrous sodium sulfate, and evaporated to obtain 4g of crude product. Purify through a column, eluent (ethyl acetate: petroleum ether = 1:20), collect the product to obtain 2.9g of oil. The yield is 80.1%, and the purity is 97.88%.

[0078] ESI-MS: m / z=282.1(M+H) + .

[0079] Step 3: Preparation of intermediate e

[0080] Take 820mg (2.211mmol) of compound d, 12ml of isopropanol, and 3ml of acetone, add 510mg (4.428mmol) of tetramethylguanidine, stir for 5 minutes, add 930mg (3.308mmol) of intermediate c, and stir at room temperature overnight. After the reaction is completed, add saturated ammonium chloride to terminate the reaction, add ethyl acetate to extract, wash the organic phase with water and saturated brine in turn, dry with anhydrous sodium sulfate, and evaporate the solvent to obtain 1.7g of crude product. Purify through a column, eluent (ethyl acetate: petroleum ether = 1:1), collect the product to obtain 1.08g of oil. The yield is 85.8% and the purity is 95.90%.

[0081] ESI-MS: m / z=571.1(M+H) + .

[0082] Step 4: Preparation of intermediate f

[0083] Take 850 mg (1.486 mmol) of compound e, add 7 ml of dichloromethane to dissolve, then add 7 ml of trifluoroacetic acid, stir and react at room temperature for 1 hour. After the reaction is complete, evaporate the solvent, dissolve the residue in dichloromethane and evaporate again, repeat this three times until the residue after evaporation becomes solid. The solid is used directly in the next step without purification.

[0084] Step 5: Preparation of title compound 1

[0085] The solid obtained in step 4 was suspended in 8 ml of tetrahydrofuran, 230 mg (1.486 mmol) of compound g was added, cooled to below 0 ° C, 494 mg (6.245 mmol) of pyridine was added, and then 1.89 g (2.97 mmol) of 1-propylphosphoric anhydride (50% ethyl acetate solution) diluted with 8 ml of tetrahydrofuran was added dropwise. After the addition, it was stirred at 0-5 ° C for 10 minutes and stirred at room temperature for 30 minutes. After the reaction was completed, water was added to terminate the reaction, EA was added for extraction, and the organic phase was washed with 5% citric acid, saturated sodium bicarbonate, water, saturated brine, dried over anhydrous sodium sulfate, and the solvent was evaporated to obtain 1.5 g of crude product. 15 ml of ethyl acetate was added to the crude product and stirred at room temperature for 2 hours, filtered, and the filter cake was washed with ethyl acetate. The filter cake was vacuum dried to obtain 783 mg of white solid, with a yield of 81.0% and a purity of 97.11%.

[0086] ESI-MS: m / z=651.1(M+H) + .

[0087] 1 H NMR (400 MHz, DMSO- d 6)δ: 10.88 (s, 1H), 9.21(s, 1H), 7.91-7.80 (m,2H), 7.77 (d, 1H), 7.73-7.64 (m, 2H), 7.51 (d, 2H), 7.36 (dd,1H), 7.11 (s,1H), 6.51 (s, 1H), 5.81 (m, 1H), 3.25 (s, 3H), 3.20 (s, 3H), 2.53-2.35 (m, 4H).

[0088] Example 2: Preparation of Compound 2

[0089]

[0090] Take 734 mg (1.127 mmol) of compound 1, add 15 ml N, N-dimethylacetamide to dissolve, add 15 ml methanol, add 142 mg (3.384 mmol) lithium hydroxide monohydrate dissolved in 15 ml water, and stir at room temperature for 1 hour. After the reaction is completed, add 5% citric acid aqueous solution to terminate the reaction, add ethyl acetate to extract, wash the organic phase with water and saturated brine in turn, dry with anhydrous sodium sulfate, and evaporate the solvent to obtain 730 mg of crude product. Purify through a column, eluent (dichloromethane: methanol = 10:1), collect the product to obtain 647 mg of solid. The yield is 90.1% and the purity is 96.05%.

[0091] ESI-MS: m / z=637.1(M+H) + .

[0092] 1 H NMR (400 MHz, DMSO- d 6 )δ: 12.71 (s, 1H),10.83 (s, 1H), 9.20 (s, 1H),7.96-7.87 (m, 2H), 7.78 (d, 1H), 7.76-7.68 (m, 2H), 7.52 (d, 2H), 7.37 (dd,1H), 7.12 (s, 1H), 6.53 (s, 1H), 5.88 (m, 1H), 3.28 (s, 3H), 2.58-2.47(m, 2H), 2.41-2.30(m, 2H).

[0093] Example 3: Preparation of Compound 3

[0094]

[0095] Take 626mg (0.982mmol) of compound 2 and dissolve it in 7ml N,N-dimethylacetamide, add 118mg (1.178mmol) of N-methylpiperazine, 448mg (1.178mmol) of HATU, cool to below 0℃, and drop 380mg (2.94mmol) of N,N-diisopropylethylamine dissolved in 7ml N,N-dimethylacetamide. After the addition, stir at 0~5℃ for 10 minutes, and stir at room temperature for 30 minutes. After the reaction is completed, add water to terminate the reaction, add EA to extract, and wash the obtained organic phase with sodium bicarbonate, water, saturated brine, dry with anhydrous sodium sulfate, and evaporate the solvent to obtain 800mg of crude product. Purify through a column, eluent (dichloromethane: methanol = 20:1), collect the product to obtain 622mg of solid. The yield is 88.1% and the purity is 99.20%.

[0096] ESI-MS: m / z=719.2(M+H) + .

[0097] 1 H NMR (400 MHz, DMSO- d 6 )δ: 10.69 (s, 1H), 9.20 (s, 1H), 7.89-7.79 (m,2H), 7.76 (d, 1H), 7.71-7.60 (m, 2H), 7.52 (d, 2H), 7.38 (dd,1H), 7.12 (s,1H), 6.54 (s, 1H), 5.53 (m, 1H), 3.47-3.33 (m, 4H), 3.25 (s, 3H), 2.45-2.19 (m, 8H), 2.17 (s, 3H).

[0098] Example 4: Preparation of Compound 4

[0099] The preparation method is the same as that of Examples 1 to 3, except that the 4-amino-2-fluorobenzamide in step 5 is replaced with an equal molar amount of 4-amino-2-fluoro-N-methylbenzamide to obtain the title compound 4 with a purity of 97.42%.

[0100] ESI-MS: m / z=733.2(M+H) + .

[0101] 1 H NMR (400 MHz, DMSO- d 6)δ: 10.23 (s, 1H), 9.19 (s, 1H), 7.87-7.76 (m,2H), 7.75 (d, 1H), 7.70-7.58 (m, 3H), 7.36 (dd, 1H), 7.10(s, 1H), 6.52 (s,1H), 5.51 (m, 1H), 3.42-3.30 (m, 4H), 3.24 (s, 3H), 2.78 (d, 3H) 2.41-2.16 (m,8H), 2.16 (s, 3H).

[0102] Example 5: Preparation of Compound 5

[0103] The preparation method is the same as that of Examples 1 to 3, except that the (R)-2-amino-5-methoxy-5-oxopentanoic acid in step 1 is replaced with an equal molar amount of (R)-2-amino-4-methoxy-4-oxobutanoic acid to obtain the title compound 5 with a purity of 98.37%.

[0104] ESI-MS: m / z=705.2(M+H) + .

[0105] 1 H NMR (400 MHz, DMSO- d 6 )δ: 10.72 (s, 1H), 9.12 (s, 1H), 7.88-7.79 (m,2H), 7.73 (t, 1H), 7.67 (t, 1H), 7.62 (dd, 1H), 7.53 (d,2H), 7.37 (dd, 1H),7.05 (s, 1H), 6.53 (s, 1H), 5.79 (t, 1H), 3.59-3.36 (m, 4H), 3.27-3.16 (m,5H), 2.37-2.18 (m, 4H), 2.16 (s, 3H).

[0106] Example 6: Preparation of Compound 6

[0107] The preparation method is the same as that of Example 3, except that the (R)-2-amino-5-methoxy-5-oxopentanoic acid in step 1 is replaced by an equal mole of (R)-2-amino-4-methoxy-4-oxobutyric acid, and the 4-amino-2-fluorobenzamide in step 5 is replaced by an equal mole of 4-amino-2-fluoro-N-methylbenzamide to obtain the title compound 6 with a purity of 99.82%.

[0108] ESI-MS: m / z=719.2(M+H)+ .

[0109] 1 H NMR (400 MHz, DMSO- d 6 )δ:10.69 (s, 1H), 9.10 (s, 1H), 8.10-7.99 (m,1H), 7.85-7.79 (m 2H), 7.73 (s, 1H), 7.67-7.59 (m, 2H), 7.37 (dd, 1H), 7.05(s, 1H), 6.53 (s, 1H), 5.79 (t, 1H), 3.56-3.40 (m, 4H), 3.25-3.12 (m, 5H), 2.76 (d, 3H), 2.36-2.19 (m, 4H), 2.16 (s, 3H).

[0110] Example 7: Preparation of Compound 7

[0111] The preparation method is the same as the preparation method of Example 3, except that the (R)-2-amino-5-methoxy-5-oxopentanoic acid in step 1 is replaced by an equal mole of (R)-2-amino-4-methoxy-4-oxobutyric acid, and the 4-amino-2-fluorobenzamide in step 5 is replaced by an equal mole of 4-amino-2-fluoro-N,N-dimethylbenzamide to obtain the title compound 7 with a purity of 98.36%.

[0112] ESI-MS: m / z=733.2(M+H) + .

[0113] 1 H NMR (400 MHz, DMSO- d 6 )δ: 10.68 (s, 1H), 9.12 (s, 1H), 8.05-7.92 (m,1H), 7.81-7.73 (m 2H), 7.65-7.54 (m, 2H), 7.36 (dd, 1H), 7.01 (s, 1H), 6.51(s, 1H), 5.66 (t, 1H), 3.51-3.39 (m, 4H), 3.25-3.11 (m, 5H), 2.91 (s, 6H), 2.33-2.16 (m, 4H), 2.15 (s, 3H).

[0114] Example 8: Preparation of Compound 8

[0115] The preparation method is the same as the preparation method of Examples 1 to 3, except that the (R)-2-amino-5-methoxy-5-oxopentanoic acid in step 1 is replaced by an equal mole of (R)-2-amino-4-methoxy-4-oxobutyric acid, and the 4-amino-2-fluorobenzamide in step 5 is replaced by an equal mole of 4-amino-2-fluoro-N-cyclopropylbenzamide to obtain the title compound 8 with a purity of 99.21%.

[0116] ESI-MS: m / z=745.2(M+H) + .

[0117] 1 H NMR (400 MHz, DMSO- d 6 )δ: 10.66 (s, 1H), 9.12 (s, 1H), 8.08-7.96 (m,1H), 7.81-7.75 (m 2H), 7.70 (s, 1H), 7.64-7.58 (m, 2H), 7.35 (dd, 1H), 7.02(s, 1H),6.51 (s, 1H), 5.72 (t, 1H), 3.55-3.39 (m, 4H), 3.26-3.11 (m, 5H),2.76 (m, 1H), 2.38-2.20 (m, 4H), 2.18 (s,3H),0.88-0.81(m,2H),0.59-0.56(m,2H).

[0118] Example 9: Preparation of Compound 9

[0119] The preparation method is the same as the preparation method of Examples 1 to 3, except that the (R)-2-amino-5-methoxy-5-oxopentanoic acid in step 1 is replaced by an equal mole of (R)-2-amino-4-methoxy-4-oxobutyric acid, and the 4-amino-2-fluorobenzamide in step 5 is replaced by an equal mole of 4-amino-2-chloro-benzamide to obtain the title compound 9 with a purity of 96.98%.

[0120] ESI-MS: m / z=721.2(M+H) + .

[0121] 1 H NMR (400 MHz, DMSO- d 6)δ: 10.70 (s, 1H), 9.10 (s, 1H), 7.86-7.75 (m,2H), 7.71 (t, 1H), 7.66 (t, 1H), 7.61 (dd, 1H), 7.52 (d, 2H), 7.36 (dd, 1H),7.05(s, 1H), 6.51 (s, 1H), 5.78 (t, 1H), 3.59-3.36 (m, 4H), 3.28-3.16 (m,5H), 2.37-2.19 (m, 4H), 2.17 (s, 3H).

[0122] Example 10: Preparation of Compound 10

[0123] The preparation method is the same as the preparation method of Examples 1 to 3, except that the (R)-2-amino-5-methoxy-5-oxopentanoic acid in step 1 is replaced by an equal molar amount of (R)-2-amino-4-methoxy-4-oxobutyric acid, the 4-amino-2-fluorobenzamide in step 5 is replaced by an equal molar amount of 4-amino-2-trifluoromethyl-benzamide, and the N-methylpiperazine in step 7 is replaced by an equal molar amount of 4-cyclopropylpiperazine to obtain the title compound 10 with a purity of 99.30%.

[0124] ESI-MS: m / z=781.2 (M+H) + .

[0125] 1 H NMR (400 MHz, DMSO- d 6 )δ: 10.80 (s, 1H), 9.13 (s, 1H), 8.12 (d, 1H), 8.02-7.72 (m, 5H), 7.53 (d, 1H), 7.14 (s, 1H), 6.54 (s, 1H), 5.51 (t, 1H),3.59-3.36(m, 4H), 3.28-3.16 (m, 5H), 2.37-2.20 (m, 4H), 2.18 (m, 1H),0.85-0.80(m,2H),0.59-0.54(m,2H).

[0126] Example 11: Preparation of Compound 11

[0127] The preparation method is the same as the preparation method of Examples 1 to 3, except that the (R)-2-amino-5-methoxy-5-oxopentanoic acid in step 1 is replaced by an equal mole of (R)-2-amino-4-methoxy-4-oxobutyric acid, and the 4-amino-2-fluorobenzamide in step 5 is replaced by an equal mole of 4-amino-2-methoxybenzamide to obtain the title compound 11 with a purity of 98.11%.

[0128] ESI-MS: m / z=717.2(M+H) + .

[0129] 1 H NMR (400 MHz, DMSO- d 6 )δ: 10.68 (s, 1H), 9.14 (d, 1H), 7.95-7.73 (m,4H), 7.56 (dd, 2H), 7.46 (d, 1H), 7.19 (dd, 1H), 7.14 (s, 1H), 6.53 (s, 1H), 5.54(t, 1H), 3.86 (s, 3H), 3.58-3.37 (m, 4H), 3.28-3.15 (m, 5H), 2.36-2.18 (m, 4H), 2.16 (s, 3H).

[0130] Example 12: Preparation of Compound 12

[0131] Preparation of intermediate (S)-4-(2-(4-(5-chloro-2-(4-(trifluoromethyl)-1H-1,2,3-triazol-1-yl)phenyl)-5-methoxy-2-oxopyridin-1(2H)-yl)-4-(4-methylpiperazine-1-yl-4-oxobutyramide)-2-methoxybenzoic acid tert-butyl ester: The preparation method is the same as the preparation method of Examples 1 to 3, except that the (R)-2-amino-5-methoxy-5-oxopentanoic acid in step 1 is replaced with an equal mole of (R)-2-amino-4-methoxy-4-oxobutyric acid, and the 4-amino-2-fluorobenzamide in step 5 is replaced with an equal mole of 4-amino-2-methoxy-benzoic acid tert-butyl ester to obtain an intermediate compound with a purity of 96.50%.

[0132] ESI-MS: m / z=774.3 (M+H) + .

[0133] Preparation of title compound 12: Take 200 mg (0.258 mmol) of (S)-4-(2-(4-(5-chloro-2-(4-(trifluoromethyl)-1H-1,2,3-triazol-1-yl)phenyl)-5-methoxy-2-oxopyridin-1(2H)-yl)-4-(4-methylpiperazin-1-yl-4-oxobutanamide)-2-methoxybenzoic acid tert-butyl ester, add 2 ml of dichloromethane to dissolve, then add 2 ml of trifluoroacetic acid, and stir at room temperature for 1 hour. After the reaction is completed, evaporate the solvent, dissolve the residue in dichloromethane and evaporate it again, and repeat this three times until the residue after evaporation becomes solid. Purify through a column, eluent (dichloromethane: methanol = 10:1), collect the target product to obtain 150 mg of pure product, with a yield of 81.0% and a purity of 97.32%.

[0134] ESI-MS: m / z=718.2(M+H) + .

[0135] 1 H NMR (400 MHz, DMSO- d 6 )δ: 12.35 (s, 1H), 10.68 (s, 1H), 9.14 (d,1H), 7.93-7.75 (m, 3H), 7.68 (d, 1H), 7.54 (d, 1H), 7.22-7.10 (m, 2H), 6.54(s, 1H), 5.54 (t, 1H), 3.78 (s, 3H), 3.60-3.39 (m, 4H), 3.28-3.17 (m, 5H), 2.38-2.20 (m, 4H), 2.18 (s, 3H).

[0136] Example 13: Preparation of Compound 13

[0137] The preparation method is the same as that of Examples 1 to 3, except that the (R)-2-amino-5-methoxy-5-oxopentanoic acid in step 1 is replaced by an equal molar amount of (R)-2-amino-4-methoxy-4-oxobutanoic acid, and the N-methylpiperazine in step 7 is replaced by an equal molar amount of morpholine to obtain the title compound 13 with a purity of 99.48%.

[0138] ESI-MS: m / z=692.1(M+H) + .

[0139] 1 H NMR (400 MHz, DMSO- d 6)δ: 10.75 (s, 1H), 9.14 (s, 1H), 7.85-7.80 (m,2H), 7.74 (s, 1H), 7.72-7.60 (m, 2H), 7.54 (d, 2H), 7.38 (dd, 1H), 7.06 (s, 1H), 6.54 (s, 1H), 5.82 (t, 1H), 3.66-3.37 (m, 8H), 3.30-3.15 (m, 5H).

[0140] Example 14: Preparation of Compound 14

[0141] The preparation method is the same as the preparation method of Examples 1 to 3, except that the (R)-2-amino-5-methoxy-5-oxopentanoic acid in step 1 is replaced by an equal molar amount of (R)-2-amino-4-methoxy-4-oxobutyric acid, the 4-amino-2-fluorobenzamide in step 5 is replaced by an equal molar amount of 4-amino-2-fluoro-N-methylbenzamide, and the N-methylpiperazine in step 7 is replaced by an equal molar amount of morpholine to obtain the title compound 14 with a purity of 98.65%.

[0142] ESI-MS: m / z=706.1(M+H) + .

[0143] 1 H NMR (400 MHz, DMSO- d 6 )δ: 10.78 (s, 1H), 9.12 (s, 1H), 7.88-7.83 (m,2H), 7.75 (s, 1H), 7.71-7.62 (m, 2H), 7.53 (s, 1H), 7.36 (dd, 1H), 7.05 (s, 1H), 6.52 (s, 1H), 5.80 (t, 1H), 3.66-3.37 (m, 8H), 3.30-3.13 (m, 5H), 2.78 (d, 3H).

[0144] Example 15: Preparation of Compound 15

[0145] The preparation method is the same as that of Examples 1 to 3, except that the (R)-2-amino-5-methoxy-5-oxopentanoic acid in step 1 is replaced by an equal molar amount of (R)-2-amino-4-methoxy-4-oxobutanoic acid, and the N-methylpiperazine in step 7 is replaced by an equal molar amount of 4-butylpiperazine to obtain the title compound 15 with a purity of 98.74%.

[0146] ESI-MS: m / z=747.3 (M+H)+ .

[0147] 1 H NMR (400 MHz, DMSO- d 6 ) δ: 10.73 (s, 1H), 9.13 (s, 1H), 7.88-7.78(m, 2H), 7.74 (t, 1H), 7.72-7.60 (m, 2H), 7.54 (d, 2H), 7.38(dd, 1H), 7.05(s, 1H), 6.54 (s, 1H), 5.79 (t, 1H), 3.52-3.39 (m, 4H), 3.30-3.11 (m, 5H), 2.43-2.14 (m, 4H), 1.49-1.15 (m, 6H), 0.87 (t, 3H).

[0148] Example 16: Preparation of Compound 16

[0149] The preparation method is the same as the preparation method of Examples 1 to 3, except that the (R)-2-amino-5-methoxy-5-oxopentanoic acid in step 1 is replaced by an equal molar amount of (R)-2-amino-4-methoxy-4-oxobutyric acid, the 4-amino-2-fluorobenzamide in step 5 is replaced by an equal molar amount of 4-amino-2-fluoro-N-methylbenzamide, and the N-methylpiperazine in step 7 is replaced by an equal molar amount of 4-butylpiperazine to obtain the title compound 16 with a purity of 97.98%.

[0150] ESI-MS: m / z=761.3 (M+H) + .

[0151] 1 H NMR (400 MHz, DMSO- d 6 ) δ: 10.78 (s, 1H), 9.12 (s, 1H), 7.89-7.79(m, 2H), 7.75 (t, 1H), 7.70-7.60 (m, 2H), 7.52 (s, 1H), 7.36(dd, 1H), 7.08(s, 1H), 6.53 (s, 1H), 5.81 (t, 1H), 3.50-3.39 (m, 4H), 3.30-3.15 (m, 5H), 2.79 (d, 3H), 2.46-2.18 (m, 4H), 1.50-1.14 (m, 6H), 0.88 (t,3H).

[0152] Example 17: Preparation of Compound 17

[0153] The preparation method is the same as that of Examples 1 to 3, except that the (R)-2-amino-5-methoxy-5-oxopentanoic acid in step 1 is replaced by an equal molar amount of (R)-2-amino-4-methoxy-4-oxobutanoic acid, and the N-methylpiperazine in step 7 is replaced by an equal molar amount of 4-cyclopropylmethylpiperazine to obtain the title compound 17 with a purity of 96.63%.

[0154] ESI-MS: m / z=745.2(M+H) + .

[0155] 1 H NMR (400 MHz, DMSO- d 6 ) δ: 10.73 (s, 1H), 9.12 (s, 1H), 7.86-7.79(m, 2H), 7.73 (t, 1H), 7.70-7.60 (m, 2H), 7.54 (d, 2H), 7.38(dd, 1H), 7.05(s, 1H), 6.54 (s, 1H), 5.79 (t, 1H), 3.54-3.37 (m, 4H), 3.30-3.09 (m, 5H), 2.47-2.25 (m, 4H), 2.19-2.14 (t, 2H), 0.87-0.79 (m, 1H),0.47-0.43 (m, 2H),0.28-0.23 (m, 2H).

[0156] Example 18: Preparation of Compound 18

[0157] The preparation method is the same as the preparation method of Examples 1 to 3, except that the (R)-2-amino-5-methoxy-5-oxopentanoic acid in step 1 is replaced by an equal molar amount of (R)-2-amino-4-methoxy-4-oxobutanoic acid, the 4-amino-2-fluorobenzamide in step 5 is replaced by an equal molar amount of 4-amino-2-fluoro-N-methylbenzamide, and the N-methylpiperazine in step 7 is replaced by an equal molar amount of 4-cyclopropylmethylpiperazine to obtain the title compound 18 with a purity of 99.13%.

[0158] ESI-MS: m / z=759.2(M+H) + .

[0159] 1 H NMR (400 MHz, DMSO- d 6) δ: 10.78 (s, 1H), 9.13 (s, 1H), 7.88-7.81(m, 2H), 7.71 (t, 1H), 7.68-7.59 (m, 2H), 7.51 (s, 1H), 7.36(dd, 1H), 7.01(s, 1H), 6.50 (s, 1H), 5.78 (t, 1H), 3.52-3.37 (m, 4H), 3.30-3.11 (m, 5H), 2.76 (d, 3H), 2.49-2.28 (m, 4H), 2.18-2.12 (t, 2H), 0.88-0.78(m, 1H), 0.48-0.45 (m, 2H), 0.28-0.23 (m, 2H).

[0160] Example 19: Preparation of Compound 19

[0161] The preparation method is the same as that of Examples 1 to 3, except that the (R)-2-amino-5-methoxy-5-oxopentanoic acid in step 1 is replaced by an equal molar amount of (R)-2-amino-4-methoxy-4-oxobutanoic acid, and the N-methylpiperazine in step 7 is replaced by an equal molar amount of thiomorpholine to obtain the title compound 19 with a purity of 98.63%.

[0162] ESI-MS: m / z=708.1(M+H) + .

[0163] 1 H NMR (400 MHz, DMSO- d 6 )δ: 10.72 (s, 1H), 9.13 (s, 1H), 7.90 - 7.79 (m, 2H), 7.74 (t, 1H), 7.68 (t, 1H), 7.62 (dd, 1H), 7.58 - 7.51 (d, 2H), 7.38(dd,1H), 7.06 (s, 1H), 6.54 (s, 1H), 5.81 (t, 1H), 3.87 - 3.58 (m, 4H), 3.31-3.17 (m, 5H), 2.64- 2.60(m, 2H), 2.53-2.51 (m, 2H).

[0164] Example 20: Preparation of Compound 20

[0165] The preparation method is the same as the preparation method of Examples 1 to 3, except that the (R)-2-amino-5-methoxy-5-oxopentanoic acid in step 1 is replaced by an equal molar amount of (R)-2-amino-4-methoxy-4-oxobutyric acid, the 4-amino-2-fluorobenzamide in step 5 is replaced by an equal molar amount of 4-amino-2-fluoro-N-methylbenzamide, and the N-methylpiperazine in step 7 is replaced by an equal molar amount of thiomorpholine to obtain the title compound 20 with a purity of 98.36%.

[0166] ESI-MS: m / z=722.2(M+H) + .

[0167] 1 H NMR (400 MHz, DMSO- d 6 )δ: 10.76 (s, 1H), 9.15 (s, 1H), 7.87-7.80 (m,2H), 7.74 (s, 1H), 7.72-7.60 (m, 2H), 7.54 (s, 1H), 7.36 (dd, 1H), 7.06 (s,1H),6.53 (s, 1H), 5.89 (t, 1H), 3.60-3.39 (m, 4H), 3.30-3.11 (m, 5H),2.79 (d,3H),2.66-2.55(m, 4H).

[0168] Example 21: Preparation of Compound 21

[0169] The preparation method is the same as the preparation method of Examples 1 to 3, except that the (R)-2-amino-5-methoxy-5-oxopentanoic acid in step 1 is replaced by an equal molar amount of (R)-2-amino-4-methoxy-4-oxobutyric acid, the 4-amino-2-fluorobenzamide in step 5 is replaced by an equal molar amount of 4-amino-pyridineamide, and the N-methylpiperazine in step 7 is replaced by an equal molar amount of pyrrole to obtain the title compound 21 with a purity of 96.82%.

[0170] ESI-MS: m / z=592.2(M+H) + .

[0171] 1 H NMR (400 MHz, DMSO- d 6)δ: 10.23 (s, 1H), 9.60 (s, 1H), 8.62 (d, 1H), 8.55-8.43 (m, 2H), 7.98 (d, 2H), 7.74 (s, 1H), 7.72-7.60 (m, 2H), 7.05 (s, 1H), 6.64 (s, 1H), 5.88 (t, 1H), 3.30-3.11 (m, 5H), 3.05-2.89 (m, 4H), 1.82-1.70 (m, 4H).

[0172] Example 22: Preparation of Compound 22

[0173] The preparation method is the same as that of Examples 1 to 3, except that 4-amino-2-fluorobenzamide in step 5 is replaced with an equal molar amount of tert-butyl 4-amino-2-fluorobenzoate to obtain the title compound 22 with a purity of 98.21%.

[0174] ESI-MS: m / z=776.3 (M+H) + .

[0175] 1 H NMR (400 MHz, DMSO- d 6 )δ: 10.81 (s, 1H), 9.22 (s, 1H), 7.92-7.73 (m,4H), 7.68 (dd, 1H), 7.42 (dd, 1H), 7.12 (s, 1H), 6.55 (s, 1H), 5.53 (m, 1H), 3.43(t, 2H), 3.35 (t, 2H), 3.24 (s, 3H), 2.46-2.06 (m, 11H), 1.53 (s, 9H).

[0176] Example 23: Preparation of Compound 23

[0177] Take 200 mg (0.258 mmol) of compound 22, add 2 ml of dichloromethane to dissolve, then add 2 ml of trifluoroacetic acid, stir at room temperature for 1 hour. After the reaction is complete, evaporate the solvent, dissolve the residue in dichloromethane and evaporate it again, repeat this three times until the residue after evaporation becomes solid. Purify by column, eluent (dichloromethane: methanol = 10:1), collect the title target to obtain 147 mg of pure product, yield 79.1%, purity: 97.86%.

[0178] ESI-MS: m / z=720.2(M+H) + .

[0179] 1 H NMR (400 MHz, DMSO- d 6 )δ: 12.88(s, 1H), 10.50 (s, 1H), 9.14 (d, 1H),8.11-7.89 (m, 3H), 7.68 (d, 1H), 7.51-7.32 (m, 2H), 7.19 (s, 1H), 6.53 (s, 1H), 5.53 (m, 1H), 3.47-3.33 (m, 4H), 3.25 (s, 3H), 2.46-2.20 (m, 8H), 2.17 (s, 3H).

[0180] Example 24: Preparation of Compound 24

[0181] The preparation method is the same as that of Example 23, except that the (R)-2-amino-5-methoxy-5-oxopentanoic acid in step 1 is replaced with an equal molar amount of (R)-2-amino-4-methoxy-4-oxobutanoic acid to obtain the title compound 24 with a purity of 97.90%.

[0182] ESI-MS: m / z=706.2(M+H) + .

[0183] 1 H NMR (400 MHz, DMSO- d 6 )δ: 12.72(s, 1H), 10.55 (s, 1H), 9.16 (d, 1H),8.02-7.81 (m, 3H), 7.65 (d, 1H), 7.50-7.31 (m, 2H), 7.15 (s, 1H), 6.51 (s, 1H), 5.58 (m, 1H), 3.55-3.36 (m, 4H), 3.29-3.12 (m, 5H), 2.39-2.17 (m, 4H), 2.18 (s, 3H).

[0184] Example 25: Preparation of Compound 25

[0185] The preparation method is the same as that of Example 23, except that the (R)-2-amino-5-methoxy-5-oxopentanoic acid in step 1 is replaced by an equal molar amount of (R)-2-amino-4-methoxy-4-oxobutanoic acid, and the N-methylpiperazine in step 7 is replaced by an equal molar amount of morpholine to obtain the title compound 25 with a purity of 98.41%.

[0186] ESI-MS: m / z=693.2(M+H) + .

[0187] 1 H NMR (400 MHz, DMSO- d 6 )δ: 12.31(s, 1H), 10.62 (s, 1H), 9.19 (d, 1H),8.00-7.78 (m, 3H), 7.68 (d, 1H), 7.53-7.35 (m, 2H), 7.15 (s, 1H), 6.53 (s, 1H), 5.59 (m, 1H), 3.68-3.37 (m, 8H), 3.30-3.11 (m, 5H).

[0188] Example 26: Preparation of Compound 26

[0189] The preparation method is the same as that of Example 23, except that the (R)-2-amino-5-methoxy-5-oxopentanoic acid in step 1 is replaced by an equal molar amount of (R)-2-amino-4-methoxy-4-oxobutanoic acid, and the N-methylpiperazine in step 7 is replaced by an equal molar amount of 4-butylpiperazine to obtain the title compound 26 with a purity of 98.82%.

[0190] ESI-MS: m / z=748.2(M+H) + .

[0191] 1 H NMR (400 MHz, DMSO- d 6 )δ: 12.78(s, 1H), 10.09 (s, 1H), 9.14 (d, 1H),7.98-7.74 (m, 3H), 7.61 (d, 1H), 7.49-7.30 (m, 2H), 7.11(s, 1H), 6.48 (s,1H),5.51 (m, 1H),3.50-3.36 (m, 4H), 3.30-3.11 (m, 5H), 2.48-2.18 (m, 4H),1.55-1.23 (m, 6H), 0.89 (t, 3H).

[0192] Example 27: Preparation of Compound 27

[0193] The preparation method is the same as that of Example 23, except that the (R)-2-amino-5-methoxy-5-oxopentanoic acid in step 1 is replaced by an equal mole of (R)-2-amino-4-methoxy-4-oxobutyric acid, and the 4-amino-2-fluorobenzoic acid tert-butyl ester in step 5 is replaced by an equal mole of 4-amino-benzoic acid tert-butyl ester to obtain the title compound 27 with a purity of 97.87%.

[0194] ESI-MS: m / z=688.2(M+H) + .

[0195] 1 H NMR (400 MHz, DMSO- d 6 )δ: 12.78 (s, 1H), 10.82 (s, 1H), 9.19 (s,1H), 8.01-7.85 (d, 2H), 7.80 (d, 2H), 7.66 (d, 1H), 7.51- 7.38 (m, 2H), 7.15(s, 1H), 6.49 (s, 1H), 5.58 (m, 1H), 3.59- 3.36 (m, 4H), 3.30-3.12 (m, 5H), 2.42-2.19 (m, 4H), 2.16 (s, 3H).

[0196] Example 28: Preparation of Compound 28

[0197] The preparation method is the same as the preparation method of Example 23, except that the (R)-2-amino-5-methoxy-5-oxopentanoic acid in step 1 is replaced by an equal mole of (R)-2-amino-4-methoxy-4-oxobutanoic acid, the 4-(5-chloro-2-(4-(trifluoromethyl)-1H-1,2,3-triazol-1-yl)phenyl)-5-methoxypyridine-2(1H)-one in step 3 is replaced by an equal mole of 4-(5-chloro-2-(4-chloro-1H-1,2,3-triazol-1-yl)phenyl)-5-methoxypyridine-2(1H)-one, and the N-methylpiperazine in step 7 is replaced by an equal mole of thiomorpholine to obtain the title compound 28 with a purity of 98.26%.

[0198] ESI-MS: m / z=675.1(M+H) + .

[0199] 1 H NMR (400 MHz, DMSO- d 6)δ: 12.89(s, 1H), 10.58 (s, 1H), 8.78 (d, 1H),8.01-7.79 (m, 3H), 7.69 (d, 1H), 7.55-7.38 (m, 2H), 7.15 (s, 1H), 6.58 (s, 1H), 5.56 (m, 1H), 3.60-3.35 (m, 4H), 3.30-3.11 (m, 5H), 2.68-2.56 (m, 4H).

[0200] Example 29: Preparation of Compound 29

[0201] The preparation method is the same as that of Example 23, except that the (R)-2-amino-5-methoxy-5-oxopentanoic acid in step 1 is replaced by an equal molar amount of (R)-2-amino-4-methoxy-4-oxobutanoic acid, the tert-butyl 4-amino-2-fluorobenzoate in step 5 is replaced by an equal molar amount of tert-butyl 4-aminobenzoate, and the N-methylpiperazine in step 7 is replaced by an equal molar amount of 4-cyclopropylmethylpiperazine to obtain the title compound 29 with a purity of 97.17%.

[0202] ESI-MS: m / z=728.2(M+H) + .

[0203] 1 H NMR (400 MHz, DMSO- d 6 )δ: 12.73 (s, 1H), 10.78 (s, 1H), 9.20 (s,1H), 8.00-7.84 (d, 2H), 7.810 (d, 2H), 7.61 (d, 1H), 7.50- 7.37 (m, 2H), 7.13(s,1H), 6.50 (s, 1H), 5.59 (m, 1H), 3.58-3.36 (m, 4H), 3.30-3.11 (m, 5H), 2.49-2.21 (m, 4H), 2.20-2.18 (t, 2H), 0.88-0.80 (m, 1H), 0.51-0.46 (m, 2H),0.28-0.23 (m, 2H).

[0204] Example 30: Preparation of Compound 30

[0205] The preparation method is the same as the preparation method of Example 23, except that the (R)-2-amino-5-methoxy-5-oxopentanoic acid in step 1 is replaced by an equal mole of (R)-2-amino-4-methoxy-4-oxobutanoic acid, the 4-(5-chloro-2-(4-(trifluoromethyl)-1H-1,2,3-triazol-1-yl)phenyl)-5-methoxypyridine-2(1H)-one in step 3 is replaced by an equal mole of 4-(5-chloro-(1H-tetrazol-1-yl)phenyl)-5-methoxypyridine-2(1H)-one, the tert-butyl 4-amino-2-fluorobenzoate in step 5 is replaced by an equal mole of tert-butyl 4-amino-picolinate, and the N-methylpiperazine in step 7 is replaced by an equal mole of pyrrole to obtain the title compound 30 with a purity of 98.28%.

[0206] ESI-MS: m / z=593.2(M+H) + .

[0207] 1 H NMR (400 MHz, DMSO- d 6 )δ: 12.88 (s, 1H), 10.81 (s, 1H), 9.65 (s,1H), 8.83 (d, 1H), 8.62-8.50 (m, 2H), 7.78 (s, 1H), 7.74-7.65 (m, 2H), 7.09 (s, 1H), 6.66 (s, 1H), 5.89 (t, 1H), 3.30-3.10 (m, 5H), 3.09-2.91 (m, 4H), 1.88-1.75 (m, 4H).

[0208] The compounds of the present invention also include intermediates produced during the synthesis of the above-mentioned example compounds, and isomers or pharmaceutically acceptable salts thereof, the structures of which are shown in Table 1 below.

[0209]

[0210]

[0211]

[0212] Comparative Example 1: Preparation of (S)-4-(2-(4-(5-chloro-2-(4-(trifluoromethyl)-1H-1,2,3-triazol-1-yl)phenyl)-5-methoxy-2-oxopyridin-1(2H)-yl)butyramido)-2-fluorobenzamide

[0213]

[0214] It was synthesized according to the method described in patent CN108026072B, with a purity of 98.5%.

[0215] ESI-MS: <h2 style=";text-align:left;direction:ltr"> m / z =593.1(M+H) + .

[0216] 1 H NMR (400 MHz, DMSO-d6)δ: 10.78 (s, 1H), 9.14 (s, 1H), 7.88-7.77(m, 3H), 7.72-7.61 (m,2H), 7.55 (d, 2H), 7.37 (dd, 1H), 7.13 (s, 1H), 6.54(s, 1H), 5.52 (dd, 1H), 3.25 (s, 3H), 2.18-2.00 (m, 2H), 0.78 (t, 3H).

[0217] Test Example 1: Determination of the anticoagulant effect of human plasma in vitro

[0218] 1. Test methods

[0219] Use sodium citrate (1:9) anticoagulant tubes to collect blood from healthy individuals, mix it thoroughly with the anticoagulant immediately, and centrifuge it at 4000r / min for 15 minutes at room temperature. After centrifugation, use a pipette to draw the plasma sample and freeze it (-80℃) for later use.

[0220] Weigh an appropriate amount of test compound and prepare it into a 10-100 mM stock solution with 100% DMSO (the specific concentration of the stock solution depends on the properties of the compound); then prepare working solutions of different concentrations using healthy human plasma as the solution (specifically: 1, 3, 10 μM), mix thoroughly; incubate at 37°C for 3 minutes and then perform APTT determination on a machine (model CS-2000I).

[0221] 2. Data processing

[0222] GraphPad Prism software was used for analysis to calculate the APTT prolongation rate at each concentration.

[0223] ⒊Test results

[0224] The results are shown in the table below. The APTT extension rates of the tested Example compounds 3, 4, 5, 6, 9, 11, 12, 13, 14, 23, 24, 25, 26, 27, 28, and 29 at 1 μM are all better than those of the compound in Comparative Example 1, and the APTT extension rates are greater than 50%. This test result estimates that the EC150 of the above invention compounds is less than 1 μM.

[0225]

[0226] Test Example 2: Determination of the activity of coagulation factor FXIa inhibitor

[0227] 1. Test samples

[0228] Example compounds 3, 4, 5, 6, 13, 14, 15, 17, 19 and the compound of Comparative Example 1.

[0229] ⒉Test steps

[0230] 1) Prepare the experimental buffer (50 mM HEPES, 5 mM KCl, 145 mM NaCl, 1 mg / ml PEG8000, pH 7.4) and equilibrate to room temperature;

[0231] 2) Prepare 10X compound working solution;

[0232] 3) Prepare 0.8 nM human FXIa working solution (2X), mix well and set aside;

[0233] 4) Add 20 μL of FXIa working solution in step 3) to all experimental wells of a 384-well plate (Coring, 3702), centrifuge at 200 g, RT, for 10 s;

[0234] 5) Add 4 μL of the compound working solution in step 2) to the corresponding experimental wells in the 384-well plate, centrifuge at 200g, RT for 10 seconds, and then incubate the working plate at 25°C for 20 minutes;

[0235] 6) Prepare 750 μM S-2366 working solution (2.5X), mix well and set aside;

[0236] 7) Add 16 μL of S-2366 working solution from step 6) to all experimental wells in the 384-well plate, centrifuge at 200g, RT for 10 seconds, and then incubate the working plate at 37°C for 45 minutes;

[0237] 8) After incubation, use EnVision to read the absorbance value at OD405nm and collect the data.

[0238] 3. Data Analysis

[0239] 1) Z' factor = 1-3*(SD Max +SD Min ) / (Mean Max -Mean Min );

[0240] 2) CV Max = (SD Max / MeanMax )*100%;

[0241] 3) CVMin = (SD Min / Mean Min )*100%;

[0242] 4) S / B = Singal / Background;

[0243] 5) Blank control: 0.1% DMSO; Positive control: Comparative Example 1;

[0244] 6) IC 50 Calculation formula: Y=Bottom + (Top-Bottom) / (1+10^((LogIC 50 -X)*HillSlope));

[0245] X: log value of compound concentration; Y: inhibition rate (%).

[0246] The concentration of the test compound is set to 5 concentrations, namely: 200nM, 40nM, 8nM, 1.6nM, 0.32nM, and IC 50 value.

[0247] 4. Test results

[0248] As shown in the following table: Under the test conditions, the example compounds of the present invention have a significant inhibitory effect on FXIa, and the inhibitory effect is similar to that of comparative example 1, especially the inhibitory effects of example compounds 5, 6, 13, 14, 15, 17, and 19 are better than those of comparative example 1.

[0249]

[0250] Test Example 3: Pharmacokinetic Study in Rats

[0251] 1. Test samples

[0252] Example Compound 5, Compound 19 and Comparative Example 1.

[0253] ⒉ Preparation method and environmental requirements of test substances

[0254] The preparation of test substances was carried out on the conventional workbench in the preparation room.

[0255] Preparation of stock solutions of the proposed method: Prepare according to the pharmaceutical preparation regulations, use methanol as solvent to prepare 1 mg / mL stock solutions of Example Compound 5, Compound 19 and Comparative Example 1, respectively.

[0256] Preparation of dosing solution for rats: 0.5% CMC-Na was used as solvent. The injection concentration was 3 mg / mL.

[0257] ⒊Test operation

[0258] (1) Dosage regimen

[0259] The experiment used 24 SD rats, which were divided into 3 groups, each group had 8 rats and half were male and half were female, namely comparative example group 1, compound group 5, and compound group 19. The injection dosage was 3 mg / kg.

[0260] ⑵ Medication and sample collection

[0261] Rats were fasted for 12 h before administration and were allowed to drink water freely. The injection dose was 3 mg / kg.

[0262] Blank blood was collected before administration, and blood was collected at predetermined time points after administration: <h2 style=";text-align:left;direction:ltr"> 2min、5min、10min、15min、30min、 <h2 style=";text-align:left;direction:ltr"> 45 min, 1 h, 2 h, 3 h, 5 h, 7 h, 24 h, About 0.5 mL of blood was collected and placed in an EDTA-K2 tube. The plasma was separated by centrifugation and stored at -80°C.

[0263] ⑶Pharmacokinetic analysis

[0264] According to the plasma concentration data of drugs, DAS 2.0 software was used to calculate the pharmacokinetic parameters.

[0265] 4. Test results

[0266] The experimental results of the pharmacokinetic study of rat injection administration are shown in the following table. Example compound 5 and compound 19 have better AUC than comparative example 1, indicating that compound 5 and compound 19 have better efficacy at the same dosage. At the same time, compound 5 and compound 19 have shorter half-lives, indicating that example compound 5 and compound 19 have the characteristics of rapid elimination compared with comparative example 1, reducing the risk of bleeding caused by long-term non-elimination of the drug during use, and the medication process is more controllable.

[0267]

[0268] The above embodiment is only one of the preferred implementation modes of the present invention and should not be used to limit the protection scope of the present invention. Any changes or modifications that are made to the main design concept and spirit of the present invention and have no substantive significance, and the technical problems they solve are still consistent with the present invention, should be included in the protection scope of the present invention.

Claims

1. A compound represented by formula (I), its stereoisomer or pharmaceutically acceptable salt: ， in, X is selected from C; R 1 Selected from (CH2) f CONR 6 R 7 , where: f is selected from 0; R 6 , R 7 The ring is closed to form a substituted six-membered heterocyclic ring, wherein the heterocyclic ring is selected from: , where R 11 is selected from methyl, ethyl, propyl or butyl; R 2 Selected from COOR 8 or CONR 9 R 10 , where: R 8 , R 9 are independently selected from hydrogen, R 10 are independently selected from hydrogen or methyl; R 3 is selected from hydrogen, fluorine or methoxy.

2. A compound, a stereoisomer or a pharmaceutically acceptable salt thereof, characterized in that: The compound is selected from the following structures: 。 3. A compound according to any one of claims 1 to 2, a stereoisomer or a pharmaceutically acceptable salt thereof, characterized in that: The hydrogen in the compound, its stereoisomer or pharmaceutically acceptable salt may be replaced by one or more deuterium.

4. Use of the compound according to any one of claims 1 to 2, its stereoisomers or pharmaceutically acceptable salts in the preparation of a medicament for treating and / or preventing diseases associated with FXIa receptors.

5. The use according to claim 4, characterized in that The disease associated with FXIa receptor is selected from cerebrovascular arterial disease and / or peripheral arterial disease.

6. The use according to claim 4, characterized in that The disease associated with FXIa receptor is selected from thromboembolism.

7. The use according to claim 4, characterized in that The disease associated with FXIa receptor is selected from transient ischemic attack, ischemic stroke, peripheral arterial occlusion, acute limb ischemia, amputation, reocclusion and restenosis after intervention or stent thrombosis.

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