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 the use of oxopyridine compounds or their salts, isomers, preparation methods and pharmaceutical compositions thereof in the preparation of drugs for treating and preventing diseases associated with FXIa receptors, especially in the treatment and prevention of 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 Phase I clinical research. Monoclonal antibodies and antisense oligonucleotides need to be injected, and 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 compounds of the present invention are novel oxopyridine compounds. Most of the compounds in the examples show good anticoagulant effects 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 is selected from substituted or unsubstituted aromatic groups, substituted or unsubstituted heterocyclic aromatic groups, or substituted or unsubstituted heterocyclic alkyl groups;

[0011] R 2 Independently selected from NH2, NHR 8 , OH or OR 8 ; Among them, R 8 Selected from alkyl or cycloalkyl;

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

[0013] R 4 Independently selected from substituted or unsubstituted tetrazole, or substituted or unsubstituted triazole.

[0014] Further, in any one of the above compounds, stereoisomers or pharmaceutically acceptable salts thereof, the R 1is selected from substituted or unsubstituted phenyl, substituted or unsubstituted pyridyl, substituted or unsubstituted 1,4-dioxanyl, substituted or unsubstituted oxazolyl, substituted or unsubstituted isoxazolyl, substituted or unsubstituted oxadiazolyl, substituted or unsubstituted pyrazolyl, or substituted or unsubstituted dihydrooxazolyl;

[0015] or / and R 2 Independently selected from NH2, NHR 8 , OH or OR 8 ; Among them, R 8 Selected from C1-C6 alkyl or C3-6 cycloalkyl;

[0016] or / and R 3 are independently selected from hydrogen, fluorine, chlorine or bromine;

[0017] or / and R 4 Selected from substituted or unsubstituted tetrazole, or substituted or unsubstituted triazole.

[0018] Further, in any one of the above compounds, stereoisomers or pharmaceutically acceptable salts thereof, X is selected from C;

[0019] or / and R 1 is selected from phenyl, pyridyl, oxadiazolyl, pyrazolyl, isoxazolyl, methyloxadiazolyl, methylpyrazolyl, methylisoxazolyl, cyclopropyloxadiazolyl, cyclopropylpyrazolyl or cyclopropylisoxazolyl;

[0020] or / and R 2 Independently selected from NH2, NHR 8 , OH or OR 8 ; Among them, R 8 Selected from methyl, ethyl, cyclopropyl, cyclopropylmethyl, tert-butyl;

[0021] or / and R 3 is selected from hydrogen, fluorine, chlorine or bromine;

[0022] or / and R 4 is selected from substituted or unsubstituted tetrazole, or substituted or unsubstituted triazole, and the substituent is selected from halogen or trifluoromethyl.

[0023] Further, in any one of the above compounds, stereoisomers or pharmaceutically acceptable salts thereof, X is selected from N;

[0024] or / and R 1 is selected from phenyl, pyridyl, oxadiazolyl, pyrazolyl, isoxazolyl, methyloxadiazolyl, methylpyrazolyl, methylisoxazolyl;

[0025] or / and R 2 Independently selected from NH2, NHR 8, OH or OR 8 ; Among them, R 8 Selected from methyl, ethyl, cyclopropyl, cyclopropylmethyl, tert-butyl;

[0026] or / and R 3 are independently selected from hydrogen;

[0027] or / and R 4 is selected from substituted or unsubstituted tetrazole, or substituted or unsubstituted triazole, and the substituent is selected from halogen or trifluoromethyl.

[0028] Furthermore, the above R 1 The substituents in the substituted or unsubstituted aromatic group, substituted or unsubstituted heterocyclic aromatic group, or substituted or unsubstituted heterocycloalkyl group are selected from alkyl, cycloalkyl, alkoxy or halogen; preferably C1-C6 alkyl, C3-C6 alkoxy, C1-C6 alkoxy, fluorine or chlorine.

[0029] Further, any one of the above compounds, stereoisomers or pharmaceutically acceptable salts thereof includes the following structure:

[0030]

[0031]

[0032]

[0033]

[0034]

[0035] Furthermore, hydrogen in the structure of any of the above compounds, stereoisomers or pharmaceutically acceptable salts thereof may be replaced by one or more deuteriums.

[0036] In another aspect, the present invention provides a method for preparing any one of the above compounds, stereoisomers or pharmaceutically acceptable salts thereof, comprising the following steps:

[0037] When R 2 For NH 2 、NHR 8 , OR 8 When, the method comprises the following steps:

[0038]

[0039] Among them, R 1 , R 3 , R 4 , R 8 has the same meaning as in any of the above definitions;

[0040] Step 1: The starting material a undergoes bromination reaction to generate intermediate b;

[0041] Step 2: Intermediate b undergoes a condensation reaction with compound c to generate intermediate d;

[0042] Step 3: The intermediate d undergoes a substitution reaction with the compound e to generate a compound of formula (I);

[0043] Or when R 2 When OH is present, the method comprises the following steps:

[0044]

[0045] Among them, R 1 , R 3 , R 4 , R 8 has the same meaning as in any of the above definitions;

[0046] Step 4: Intermediate f undergoes hydrolysis under alkaline conditions to generate a compound of formula (I).

[0047] In a third aspect, the present invention provides use of any one of the above compounds, stereoisomers or pharmaceutically acceptable salts thereof in the preparation of drugs for treating and / or preventing diseases associated with FXIa receptors.

[0048] Furthermore, the above-mentioned disease associated with FXIa receptor is selected from cerebrovascular arterial disease and / or peripheral arterial disease.

[0049] Furthermore, the above-mentioned cerebrovascular arterial diseases include but are not limited to transient ischemic attack (TIA), ischemic stroke or events of thrombotic and / or thromboembolic origin leading to stroke or TIA; the above-mentioned peripheral arterial diseases include but are not limited to 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.

[0050] Furthermore, the above-mentioned ischemic stroke includes but is not limited to cardiogenic stroke, non-cardiogenic stroke, stroke caused by large artery or small artery disease, stroke caused by undetermined cause, cryptogenic stroke, embolic stroke or embolic stroke of undetermined source.

[0051] Furthermore, the aforementioned cardiogenic stroke includes but is not limited to stroke caused by atrial fibrillation; the aforementioned non-cardiogenic stroke includes but is not limited to lacunar stroke.

[0052] Beneficial effects: Compared with the prior art, the compounds of the present invention have better FXIa inhibition and in vitro human blood anticoagulant effect, and the pharmacokinetics in rats show good pharmacokinetic properties. They have the characteristics of rapid elimination, avoiding the risk of bleeding caused by long-term effects, and are therefore safer. DETAILED DESCRIPTION

[0053] The present invention will be further described in detail below in conjunction with embodiments and experimental examples. The embodiments and experimental 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.

[0054] 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.

[0055] The structure of the compound is determined by NMR ( 1 H NMR) or liquid chromatography-mass spectrometry (LC-MS).

[0056] The liquid chromatography-mass spectrometer (LC-MS) was Agilent G6120B (used with Agilent 1260 liquid chromatography); the nuclear magnetic resonance instrument ( 1 HNMR) is BrukerAVANCE-400 or BrukerAVANCE-800, NMR ( 1 H NMR) shifts (δ) are given in parts per million (ppm) using DMSO as the solvent and tetramethylsilane (TMS) as the internal standard. Chemical shifts are expressed in 10 -6 (ppm) is given as the unit.

[0057] The term "room temperature" in the present invention means a temperature between 10 and 30°C.

[0058] 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)-3-phenylpropionamido)benzoic acid methyl ester (Compound 1)

[0059]

[0060] Step 1: (R)-2-Bromo-3-phenylpropionic acid

[0061] Take 8.3g (84.69mmol) of concentrated sulfuric acid, add it to 50ml of water, cool it to below 5°C, add 14.4g (121.0mmol) of potassium bromide, 5g (31.03mmol) of D-phenylalanine, and dropwise add 3.13g (45.36mmol) of sodium nitrite dissolved in 25ml of water, control the temperature below 0-5°C, and add it in about 30 minutes. After the addition, stir and react at 0-5°C overnight. The next day, the reaction solution was extracted twice with ethyl acetate, the organic phases were combined, the organic phases were washed with water, saturated brine, dried over anhydrous sodium sulfate, and the solvent was evaporated to obtain 7g of oily substance, which was directly used in the next step without purification.

[0062] Step 2: Preparation of (R)-methyl 4-(2-bromo-3-phenylpropionamido)benzoate

[0063] Take 682mg (2.977mmol) (R) -2-bromo-3-phenylpropionic acid, dissolve it in 4ml tetrahydrofuran, add 300mg (1.985mmol) 4-aminobenzoic acid methyl ester, cool to below 0℃, add 695mg (8.331mmol) pyridine, then drop 2.53g (3.976mmol) 1-propylphosphoric anhydride (50% ethyl acetate solution) diluted with 2ml tetrahydrofuran, after adding, stir at 0~5℃ for 10 minutes, stir at room temperature for 30 minutes. After the reaction is completed, add water to terminate the reaction, add EA to extract, wash the organic phase with 5% citric acid, saturated sodium bicarbonate, water, saturated salt water, dry with anhydrous sodium sulfate, evaporate the solvent to obtain 780mg crude product. Add 5ml ethyl acetate to the crude product and stir at room temperature for 2 hours, filter, wash the filter cake with ethyl acetate, and dry the filter cake in vacuum to obtain 530mg white solid, with a yield of 73.7% and a purity of 97.11%.

[0064] ESI-MS: m / z = 362.0 (M+H) + .

[0065] Step 3: Preparation of (S)-methyl 4-(2-(4-(5-chloro-2-(4-(trifluoromethyl)-1H-1,2,3-triazol-1-yl)phenyl)-5-methoxy-2-oxopyridin-1(2H)yl)-3-phenylpropionamido)benzoate

[0066] Take 250mg (0.674mmol) 4-(5-chloro-2-(4-(trifluoromethyl)-1H-1,2,3-triazol-1-yl)phenyl)-5-methoxypyridin-2(1H)-one, 5ml isopropanol, 2ml acetone, add 233mg (2.023mmol) tetramethylguanidine, stir for 5 minutes, add 293mg (0.809mmol) (R)-4-(2-bromo-3-phenylpropionamido)benzoic acid methyl ester, stir at room temperature overnight. After the reaction is complete, 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, evaporate the solvent to obtain 600mg crude product. Purify through a column, eluent (ethyl acetate: petroleum ether = 2:1), collect the product to obtain 290mg white solid. The yield is 66.0%, and the purity is 97.90%.

[0067] ESI-MS: m / z = 652.0 (M+H) + .

[0068] 1 H NMR(400MHz,DMSO-d6)δ:10.86(s,1H),9.19(s,1H),7.95(d,2H),7.85–7.73(m,4H),7.69(s,1H),7.34– 7.24(m,3H),7.23–7.13(m,3H),6.42(s,1H),5.95(dd,1H),3.83(s,3H),3.52–3.39(m,2H),3.26(s,3H).

[0069] Example 2: 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)-3-phenylpropionamido)benzoic acid (Compound 2)

[0070]

[0071] Take 290 mg (0.383 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)-3-phenylpropionamido)benzoic acid methyl ester, add 5 ml of methanol, heat to 60°C, add 49 mg (1.168 mmol) of lithium hydroxide monohydrate dissolved in 2.5 ml of water, and stir at 60°C for 1 hour. After the reaction is complete, add 5% citric acid aqueous solution to terminate the reaction, add ethyl acetate for extraction, wash the organic phase with water and saturated brine in turn, dry over anhydrous sodium sulfate, and evaporate the solvent to obtain 250 mg of crude product. Add 5 ml of ethyl acetate and stir at room temperature for 2 hours, filter, wash the filter cake with ethyl acetate, and dry the filter cake in vacuum to obtain 200 mg of white solid, with a yield of 81.7% and a purity of 97.92%.

[0072] ESI-MS: m / z = 638.0 (M+H) + .

[0073] 1 H NMR(400MHz,DMSO-d6)δ:12.78(s,1H),10.82(s,1H),9.19(s,1H),8.01–7.85(d,2H),7.80(d,2H),7.78–7.71(d,2 H),7.69(s,1H),7.36–7.23(m,3H),7.24–7.12(m,3H),6.42(s,1H),5.96(dd,1H),3.54–3.36(m,2H),3.26(s,3H).

[0074] Example 3: Preparation of (S)-5-(2-(4-(5-chloro-2-(4-(trifluoromethyl)-1H-1,2,3-triazol-1-yl)phenyl)-5-methoxy-2-oxopyridine-1(2H)yl)-3-phenylpropionamido)picolinic acid methyl ester (Compound 3)

[0075]

[0076] The preparation method is the same as the preparation method of Example 1, except that 4-aminobenzoic acid methyl ester in step 2 is replaced by 5-aminopyridine-2-carboxylic acid methyl ester to obtain the title compound (S)-5-(2-(4-(5-chloro-2-(4-(trifluoromethyl)-1H-1,2,3-triazol-1-yl)phenyl)-5-methoxy-2-oxopyridine-1(2H)yl)-3-phenylpropionamido)picolinic acid methyl ester with a purity of 98.15%.

[0077] ESI-MS: m / z = 653.0 (M+H) + .

[0078] 1 H NMR(400MHz,DMSO-d6)δ:11.02(s,1H),9.19(s,1H),8.68(d,1H),8.22(dd,1H),7.98(d,1H),7.73(d,2H),7.69(s, 1H),7.36–7.26(m,3H),7.21–7.12(m,3H),6.42(s,1H),5.98(dd,1H),3.81(s,3H),3.59–3.44(m,2H),3.24(s,3H).

[0079] Example 4: Preparation of (S)-5-(2-(4-(5-chloro-2-(4-(trifluoromethyl)-1H-1,2,3-triazol-1-yl)phenyl)-5-methoxy-2-oxopyridine-1(2H)yl)-3-phenylpropionamido)picolinic acid (Compound 4)

[0080]

[0081] Take 200 mg (0.306 mmol) of (S)-5-(2-(4-(5-chloro-2-(4-(trifluoromethyl)-1H-1,2,3-triazol-1-yl)phenyl)-5-methoxy-2-oxopyridin-1(2H)yl)-3-phenylpropionamido)picolinic acid methyl ester, add 2 ml N,N-dimethylacetamide to dissolve, add 2 ml methanol, cool to 0°C, add 39 mg (0.929 mmol) lithium hydroxide monohydrate dissolved in 2 ml water, stir at 0-5°C for 1 hour. After the reaction is complete, 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 200 mg of crude product. Add 2 ml ethyl acetate and 2 ml methanol and stir at room temperature for 2 hours, filter, wash the filter cake with ethyl acetate, and dry the filter cake in vacuum to obtain 100 mg of white solid, with a yield of 51.3% and a purity of 97.25%.

[0082] ESI-MS: m / z = 639.0 (M+H) + .

[0083] 1H NMR(400MHz,DMSO-d6)δ:13.00(s,1H),11.40(s,1H),9.19(s,1H),8.97(d,1H),8.27(dd,1H),8.04(d,1H) ,7.80(d,2H),7.68(s,1H),7.36–7.12(m,6H),6.42(s,1H),5.98(dd,1H),3.59–3.43(m,2H),3.25(s,3H).

[0084] Example 5: 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)-3-(pyridin-4-yl)propionamido)benzoic acid methyl ester (Compound 5)

[0085]

[0086] The preparation method is the same as the preparation method of Example 1, except that D-phenylalanine is replaced by D-3-(4-pyridyl)-alanine to obtain the title compound (S)-4-(2-(4-(5-chloro-2-(4-(trifluoromethyl)-1H-1,2,3-triazol-1-yl)phenyl)-5-methoxy-2-oxopyridine-1(2H)-yl)-3-(pyridin-4-yl)propionamido)benzoic acid methyl ester with a purity of 98.26%.

[0087] ESI-MS: m / z = 653.1 (M+H) + .

[0088] 1 H NMR(400MHz,DMSO-d6)δ:10.88(s,1H),9.11(s,1H),8.71(d,2H),7.95(d,2H),7.85–7.73(m,4H),7.6 9(s,1H),7.34–7.24(m,3H),6.42(s,1H),5.95(dd,1H),3.83(s,3H),3.52–3.39(m,2H),3.26(s,3H).

[0089] Example 6: 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)-3-(pyridin-4-yl)propionamido)benzoic acid (Compound 6)

[0090]

[0091] Take 200 mg (0.306 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)-3-(pyridin-4-yl)propionamide)benzoic acid methyl ester, add 2 ml of N,N-dimethylacetamide to dissolve, add 2 ml of methanol, cool to 0°C, add 39 mg (0.929 mmol) of lithium hydroxide monohydrate dissolved in 2 ml of water, stir at 0-5°C to react for 1 hour. After the reaction is complete, add water to terminate the reaction, adjust the pH to 6-7 with 5% citric acid aqueous solution, add ethyl acetate to extract, wash the organic phase with water and saturated brine in turn, dry over anhydrous sodium sulfate, and evaporate the solvent to obtain 200 mg of crude product. 2 ml of ethyl acetate and 2 ml of methanol were added, 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 115 mg of white solid with a yield of 58.8% and a purity of 96.39%.

[0092] ESI-MS: m / z = 639.1 (M+H) + .

[0093] 1 H NMR(400MHz,DMSO-d6)δ:12.88(s,1H),11.02(s,1H),9.22(s,1H),8.55(d,2H),7.91(d,2H),7.82(d,2H) ,7.65(s,1H),7.41–7.33(m,3H),7.22(d,2H),6.40(s,1H),5.99(dd,1H),3.58–3.40(m,2H),3.28(s,3H).

[0094] Example 7: 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)-3-(5-methylisoxazol-3-yl)propionamido)-2-fluorobenzoic acid methyl ester (Compound 7)

[0095]

[0096] The preparation method is the same as the preparation method of Example 1, except that D-phenylalanine in step 1 is replaced by (R)-2-amino-3-(5-methylisoxazol-3-yl)propionic acid to obtain the title compound (S)-4-(2-(4-(5-chloro-2-(4-(trifluoromethyl)-1H-1,2,3-triazol-1-yl)phenyl)-5-methoxy-2-oxopyridine-1(2H)yl)-3-(5-methylisoxazol-3-yl)propionamido)-2-fluorobenzoic acid methyl ester with a purity of 98.85%.

[0097] ESI-MS: m / z = 675.1 (M+H) + .

[0098] 1 H NMR(400MHz,DMSO-d6)δ:10.81(s,1H),9.18(s,1H),7.78(d,2H),7.69(s,1H),7.62(t,1H),7.48(dd,1H),7.37– 7.25(m,2H),6.42(s,1H),6.10(s,1H),5.88(dd,1H),4.12(s,3H),3.26(s,3H),3.21-2.98(m,2H),2.43(s,3H).

[0099] Example 8: 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)-3-(5-methylisoxazol-3-yl)propionamido)-2-fluorobenzoic acid (Compound 8)

[0100]

[0101] The preparation method is the same as that of Example 2, except that (S)-4-(2-(4-(5-chloro-2-(4-(trifluoromethyl)-1H-1,2,3-triazol-1-yl)phenyl)-5-methoxy-2-oxopyridine-1(2H)yl)-3-(5-methylisoxazol-3-yl)propionamido)-2-fluorobenzoic acid methyl ester is used as a raw material to obtain the title compound (S)-4-(2-(4-(5-chloro-2-(4-(trifluoromethyl)-1H-1,2,3-triazol-1-yl)phenyl)-5-methoxy-2-oxopyridine-1(2H)yl)-3-(5-methylisoxazol-3-yl)propionamido)-2-fluorobenzoic acid with a purity of 98.76%.

[0102] ESI-MS: m / z = 661.1 (M+H) + .

[0103] 1H NMR(400MHz,DMSO-d6)δ:10.88(s,1H),9.19(s,1H),7.88(d,2H),7.71(s,1H),7.62(t,1H),7.48(dd,1H) ,7.39–7.28(m,2H),6.45(s,1H),6.12(s,1H),5.91(dd,1H),3.29(s,3H),3.22-3.00(m,2H),2.45(s,3H).

[0104] Example 9: 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)-3-(1-methyl-1H-pyrazol-3-yl)propionamido)benzoic acid methyl ester (Compound 9)

[0105]

[0106] The preparation method is the same as the preparation method of Example 1, except that D-phenylalanine in step 1 is replaced by (R)-2-amino-3-(1-methyl-1H-pyrazol-3-yl)propionic acid to obtain the title compound (S)-4-(2-(4-(5-chloro-2-(4-(trifluoromethyl)-1H-1,2,3-triazol-1-yl)phenyl)-5-methoxy-2-oxopyridine-1(2H)-yl)-3-(1-methyl-1H-pyrazol-3-yl)propionamido)benzoic acid methyl ester with a purity of 98.90%.

[0107] ESI-MS: m / z = 656.2 (M+H) + .

[0108] 1 H NMR(400MHz,DMSO-d6)δ:11.12(s,1H),9.20(s,1H),7.98(d,2H),7.96(d,1H),7.87–7.75(m,4H),7.68(s,1H), 7.30(d,1H),6.46(s,1H),6.22(d,1H),5.99(dd,1H),4.23(s,3H),3.83(s,3H),3.66–3.42(m,2H),3.28(s,3H).

[0109] Example 10: 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)-3-(1-methyl-1H-pyrazol-3-yl)propionamido)benzoic acid (Compound 10)

[0110]

[0111] The preparation method is the same as that of Example 2, except that (S)-4-(2-(4-(5-chloro-2-(4-(trifluoromethyl)-1H-1,2,3-triazol-1-yl)phenyl)-5-methoxy-2-oxopyridine-1(2H)-yl)-3-(1-methyl-1H-pyrazol-3-yl)propionamido)benzoic acid methyl ester is used as a raw material to obtain the title compound (S)-4-(2-(4-(5-chloro-2-(4-(trifluoromethyl)-1H-1,2,3-triazol-1-yl)phenyl)-5-methoxy-2-oxopyridine-1(2H)-yl)-3-(1-methyl-1H-pyrazol-3-yl)propionamido)benzoic acid with a purity of 99.10%.

[0112] ESI-MS: m / z = 642.1 (M+H) + .

[0113] 1 H NMR(400MHz,DMSO-d6)δ:12.66(s,1H),11.00(s,1H),9.19(s,1H),7.98(d,2H),7.96(d,1H),7.85–7.71(m,4H), 7.69(s,1H),7.30(d,1H),6.46(s,1H),6.24(d,1H),5.99(dd,1H),4.31(s,3H),3.64–3.39(m,2H),3.28(s,3H).

[0114] Example 11: Preparation of (S)-4-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-5-methoxy-2-oxopyridin-1(2H)-yl)-3-phenylpropionamido)benzoic acid (Compound 11)

[0115]

[0116] The preparation method is the same as the preparation method of Example 1 and Example 2, except that 4-(5-chloro-2-(4-(trifluoromethyl)-1H-1,2,3-triazol-1-yl)phenyl)-5-methoxypyridin-2(1H)-one is replaced with 4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-5-methoxypyridin-2(1H)-one to obtain the title compound (S)-4-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-5-methoxy-2-oxopyridine-1(2H)-yl)-3-phenylpropionamido)benzoic acid with a purity of 98.28%.

[0117] ESI-MS: m / z = 571.1 (M+H) + .

[0118] 1 H NMR(400MHz,DMSO-d6)δ:12.70(s,1H),10.53(s,1H),9.56(s,1H),8.00–7.84(d,2H),7.78(d,2H),7.75–7.69(d,2 H),7.65(s,1H),7.33–7.20(m,3H),7.14–7.00(m,3H),6.39(s,1H),5.86(dd,1H),3.51–3.45(m,2H),3.29(s,3H).

[0119] Example 12: Preparation of (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-5-methoxy-2-oxopyridin-1(2H)-yl)-3-(pyridin-4-yl)propionamido)picolinic acid (Compound 12)

[0120]

[0121] The preparation method is the same as the preparation method of Example 1 and Example 6, except that 4-(5-chloro-2-(4-(trifluoromethyl)-1H-1,2,3-triazol-1-yl)phenyl)-5-methoxypyridin-2(1H)-one is replaced by 4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-5-methoxypyridin-2(1H)-one, and the 4-aminobenzoic acid methyl ester in step 2 is replaced by 5-aminopyridine-2-carboxylic acid methyl ester to obtain the title compound (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-5-methoxy-2-oxopyridine-1(2H)-yl)-3-(pyridin-4-yl)propionylamino)picolinic acid with a purity of 97.75%.

[0122] ESI-MS: m / z = 573.1 (M+H) + .

[0123] 1 H NMR(400MHz,DMSO-d6)δ:12.88(s,1H),11.22(s,1H),9.19(s,1H),8.91(d,1H),8.67(d,2H),8.26(dd,1H),8.00 (d,1H),7.81(d,2H),7.65(s,1H),7.39–7.22(m,3H),6.37(s,1H),5.91(dd,1H),3.60–3.46(m,2H),3.28(s,3H).

[0124] Example 13: 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)-3-phenylpropanamide)benzamide (Compound 13)

[0125]

[0126] The preparation method is the same as the preparation method of Example 1, except that methyl 4-aminobenzoate in step 2 is replaced by p-aminobenzamide to obtain the title compound (S)-4-(2-(4-(5-chloro-2-(4-(trifluoromethyl)-1H-1,2,3-triazol-1-yl)phenyl)-5-methoxy-2-oxopyridine-1(2H)-yl)-3-phenylpropionamido)benzamide with a purity of 98.61%.

[0127] ESI-MS: m / z = 637.0 (M+H) + .

[0128] 1 H NMR(400MHz,DMSO-d6)δ:10.21(s,1H),8.13(s,1H),7.80(d,2H),7.68–7.55(m,4H),7.52(d,2H),7.3 4(dd,1H),7.25(m,3H),7.20–7.08(m,3H),6.42(s,1H),5.88(dd,1H),3.49–3.34(m,2H),3.25(s,3H).

[0129] Example 14: 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)-3-phenylpropionamido)-2-fluorobenzamide (Compound 14)

[0130]

[0131] The preparation method is the same as the preparation method of Example 1, except that 4-aminobenzoic acid methyl ester in step 2 is replaced by 4-amino 2-fluoroaniline to obtain the title compound (S)-4-(2-(4-(5-chloro-2-(4-(trifluoromethyl)-1H-1,2,3-triazol-1-yl)phenyl)-5-methoxy-2-oxopyridine-1(2H)-yl)-3-phenylpropionamido)-2-fluorobenzamide with a purity of 98.81%.

[0132] ESI-MS: m / z = 655.0 (M+H) + .

[0133] 1 H NMR(400MHz,DMSO-d6)δ:10.88(s,1H),9.18(s,1H),7.80(d,2H),7.74–7.61(m,3H),7.56(d,2H),7.3 7(dd,1H),7.28(m,3H),7.23–7.10(m,3H),6.43(s,1H),5.91(dd,1H),3.52–3.36(m,2H),3.26(s,3H).

[0134] Example 15: 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)-3-phenylpropionamido)-2-fluorobenzoic acid (Compound 15)

[0135]

[0136] The preparation method is the same as the preparation method of Example 1 and Example 2, except that the 4-aminobenzoic acid methyl ester in step 2 is replaced by 4-amino-2-fluorobenzoic acid methyl ester to obtain the title compound (S)-4-(2-(4-(5-chloro-2-(4-(trifluoromethyl)-1H-1,2,3-triazol-1-yl)phenyl)-5-methoxy-2-oxopyridine-1(2H)-yl)-3-phenylpropionamido)-2-fluorobenzoic acid with a purity of 97.60%.

[0137] ESI-MS: m / z = 656.0 (M+H) + .

[0138] 1 H NMR(400MHz,DMSO-d6)δ:10.81(s,1H),9.18(s,1H),7.80(d,2H),7.69(s,1H),7.61(t,1H), 7.46(dd,1H),7.38–7.12(m,7H),6.41(s,1H),5.94(dd,1H),3.48-3.39(m,3H),3.26(s,3H).

[0139] Example 16: Preparation of (S)-5-(2-(4-(5-chloro-2-(4-(trifluoromethyl)-1H-1,2,3-triazol-1-yl)phenyl)-5-methoxy-2-oxopyridin-1(2H)-yl)-3-phenylpropionamido)picolinamide (Compound 16)

[0140]

[0141] The preparation method is the same as the preparation method of Example 1, except that 4-aminobenzoic acid methyl ester in step 2 is replaced by 5-amino-2-pyridinecarboxamide to obtain the title compound (S)-5-(2-(4-(5-chloro-2-(4-(trifluoromethyl)-1H-1,2,3-triazol-1-yl)phenyl)-5-methoxy-2-oxopyridine-1(2H)-yl)-3-phenylpropionamido)picolinamide with a purity of 98.14%.

[0142] ESI-MS: m / z = 638.0 (M+H) + .

[0143] 1 H NMR(400MHz,DMSO-d6)δ:10.99(s,1H),9.25(s,1H),8.87(d,1H)8.63(dd,1H),8.51(d,1H),7.92(d,2H),7.58(d ,2H),7.42(dd,1H),7.33(m,3H),7.26–7.13(m,3H),6.55(s,1H),5.97(dd,1H),3.57–3.46(m,2H),3.28(s,3H).

[0144] Example 17: 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)-3-phenylpropionamido)-2-fluoro-N-methylbenzamide (Compound 17)

[0145]

[0146] The preparation method is the same as the preparation method of Example 1, except that the methyl 4-aminobenzoate in step 2 is replaced by N-methyl-2-fluoro-4-aminobenzamide to obtain the title compound (S)-4-(2-(4-(5-chloro-2-(4-(trifluoromethyl)-1H-1,2,3-triazol-1-yl)phenyl)-5-methoxy-2-oxopyridine-1(2H)-yl)-3-phenylpropionamido)-2-fluoro-N-methylbenzamide with a purity of 98.19%.

[0147] ESI-MS: m / z = 669.0 (M+H) + .

[0148] 1H NMR(400MHz,DMSO-d6)δ:11.08(s,1H),9.08(s,1H),8.33(m,1H),7.66–7.52(m,3H),7.48(d,2H),7.35(dd,1 H),7.26(m,3H),7.20–7.05(m,3H),6.40(s,1H),5.85(dd,1H),3.54–3.39(m,2H),3.22(s,3H),2.76(d,3H).

[0149] Example 18: Preparation of (S)-5-(2-(4-(5-chloro-2-(4-(trifluoromethyl)-1H-1,2,3-triazol-1-yl)phenyl)-5-methoxy-2-oxopyridin-1(2H)-yl)-3-phenylpropionamido)-N-propylpicolinamide (Compound 18)

[0150]

[0151] The preparation method is the same as the preparation method of Example 1, except that 4-aminobenzoic acid methyl ester in step 2 is replaced by 5-amino-N-propylpicolinamide to obtain the title compound (S)-5-(2-(4-(5-chloro-2-(4-(trifluoromethyl)-1H-1,2,3-triazol-1-yl)phenyl)-5-methoxy-2-oxopyridine-1(2H)-yl)-3-phenylpropionamido)-N-propylpicolinamide with a purity of 98.10%.

[0152] ESI-MS: m / z = 680.2 (M+H) + .

[0153] 1 H NMR(400MHz,DMSO-d6)δ:11.12(s,1H),9.21(s,1H),9.13(m,1H),8.71(d,1H)8.53(dd,1H),8.32(d,1H),7.55(d,2H),7.40(dd,1 H),7.29(m,3H),7.22(m,3H),6.50(s,1H),5.86(dd,1H),3.52–3.40(m,2H),3.28(s,3H),3.22(m,2H),1.68(m,2H),0.99(m,3H).

[0154] Example 19: Preparation of (S)-tert-butyl 4-(2-(4-(5-chloro-2-(4-(trifluoromethyl)-1H-1,2,3-triazol-1-yl)phenyl)-5-methoxy-2-oxopyridin-1(2H)-yl)-3-phenylpropionamido)-2-fluorobenzoate (Compound 19)

[0155]

[0156] The preparation method is the same as the preparation method of Example 1, except that the 4-aminobenzoic acid methyl ester in step 2 is replaced by 4-amino 2-fluorobenzoic acid tert-butyl ester to obtain the title compound (S)-4-(2-(4-(5-chloro-2-(4-(trifluoromethyl)-1H-1,2,3-triazol-1-yl)phenyl)-5-methoxy-2-oxopyridine-1(2H)-yl)-3-phenylpropionamido)-2-fluorobenzoic acid tert-butyl ester with a purity of 97.15%.

[0157] ESI-MS: m / z = 712.0 (M+H) + .

[0158] 1 H NMR(400MHz,DMSO-d6)δ:10.81(s,1H),9.18(s,1H),7.80(d,2H),7.69(s,1H),7.61(t,1H), 7.46(dd,1H),7.38–7.12(m,7H),6.41(s,1H),5.94(dd,1H),3.48-3.39(m,3H),3.26(s,3H).

[0159] Example 20: 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)-3-phenylpropionamido)-2-fluorobenzoic acid methyl ester (Compound 20)

[0160]

[0161] The preparation method is the same as the preparation method of Example 1, except that the 4-aminobenzoic acid methyl ester in step 2 is replaced by 4-amino-2-fluorobenzoic acid methyl ester to obtain the title compound (S)-4-(2-(4-(5-chloro-2-(4-(trifluoromethyl)-1H-1,2,3-triazol-1-yl)phenyl)-5-methoxy-2-oxopyridine-1(2H)-yl)-3-phenylpropionamido)-2-fluorobenzoic acid methyl ester with a purity of 96.70%.

[0162] ESI-MS: m / z = 670.0 (M+H) + .

[0163] 1H NMR(400MHz,DMSO-d6)δ:10.81(s,1H),9.18(s,1H),7.80(d,2H),7.69(s,1H),7.61(t,1H),7.46(d d,1H),7.38–7.12(m,7H),6.41(s,1H),5.94(dd,1H),3.83(s,3H),3.48-3.39(m,3H),3.26(s,3H).

[0164] 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

[0165]

[0166] It was synthesized according to the method described in patent CN108026072B, with a purity of 98.5%.

[0167] ESI-MS: m / z = 593.0 (M+H) + .

[0168] 1 H NMR(400MHz,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).

[0169] Test Example 1: Determination of the activity of coagulation factor FXIa inhibitor (50 nM inhibition rate of coagulation factor FXIa)

[0170] 1. Test samples

[0171] Example compounds 1 to 20 and comparative example 1.

[0172] ⒉Test steps

[0173] 1) Prepare experimental buffer (50 mM H1) Prepare experimental buffer (50 mM HEPES, 5 mM KCl, 145 mM NaCl, 1 mg / ml PEG 8000, pH 7.4) and equilibrate to room temperature.

[0174] 2) Prepare 10X compound working solution.

[0175] 3) Prepare 0.8 nM Human FXIa working solution (2X), mix well and set aside.

[0176] 4) Add 20 μL of the FXIa working solution prepared in step 3) to all experimental wells of a 384-well plate (Coring, 3702), and centrifuge at 200 g, RT, for 10 s.

[0177] 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.

[0178] 6) Prepare 750 μM S-2366 working solution (2.5X), mix well and set aside.

[0179] 7) Add 16 μL of the S-2366 working solution in step 6) to all experimental wells in the 384-well plate, centrifuge at 200 g, RT for 10 s, and then incubate the working plate at 37° C. for 45 min.

[0180] 8) After the incubation is completed, use EnVision to read the absorbance value at OD405nm and collect the data.

[0181] 3. Data Analysis

[0182] 1) Z'factor = 1-3*(SD Max +SD Min ) / (Mean Max -Mean Min );

[0183] 2)CV Max =(SD Max / Mean Max )*100%;

[0184] 3) CVMin = (SD Min / Mean Min )*100%;

[0185] 4) S / B = Singal / Background;

[0186] 5) Blank control: 0.1% DMSO; Positive control: Comparative Example 1;

[0187] 6) IC 50 Calculation formula: Y = Bottom + (Top-Bottom) / (1 + 10^((LogIC 50 -X)*HillSlope)).

[0188] X: log value of compound concentration; Y: Inhibition%.

[0189] 4. Test results

[0190] As shown in the following table: At 50 nM, most of the compounds of the present invention have stronger inhibitory effects on FXIa than Comparative Example 1.

[0191] Table 1 Results of activity determination of coagulation factor FXIa inhibitor

[0192] sample FXIa inhibition rate (%) sample FXIa inhibition rate (%) Compound 1 47.2 Compound 2 100.4 Compound 3 55.9 Compound 4 89.2 Compound 5 70.8 Compound 6 98.4 Compound 7 60.6 Compound 8 99.0 Compound 9 65.4 Compound 10 93.1 Compound 1 1 95.3 Compound 12 92.6 Compound 13 88.7 Compound 14 97.9 Compound 15 96.1 Compound 16 90.5 Compound 17 87.6 Compound 18 73.1 Compound 19 90.1 Compound 20 89.2 Comparative Example 1 80.5

[0193] Test Example 2: Determination of the activity of coagulation factor FXIa inhibitor (IC 50 )

[0194] The inhibitory effects of compounds 2, 4, 6, 8, 11, 14, 15, 16 and the compound of comparative example 1 on human coagulation factor FXIa were determined by in vitro enzymatic assay, and 5 concentrations, namely 200 nM, 40 nM, 8 nM, 1.6 nM and 0.32 nM, were set to detect IC50 values.

[0195] The test reagents and operating methods are as described in Test Example 1.

[0196] The test results are shown in the following table: Compounds 2, 4, 6, 8, 11, 14, 15 and 16 of the present invention have stronger inhibitory effects on FXIa than the compound of comparative example 1.

[0197] Table 2 Results of activity determination (IC50) of coagulation factor FXIa inhibitor

[0198] sample <![CDATA[IC 50 (nM)]]> sample <![CDATA[IC 50 (nM)]]> Compound 2 0.5 Compound 4 4.6 Compound 6 2.0 Compound 8 3.3 Compound 11 3.8 Compound 14 1.1 Compound 15 4.2 Compound 16 3.2 Comparative Example 1 7.59

[0199] Test Example 3: Determination of the anticoagulant effect of human plasma in vitro

[0200] 1. Test samples

[0201] Example compounds 2, 4, 14 and comparative example 1.

[0202] 2. Test methods

[0203] 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.

[0204] Weigh an appropriate amount of compound and prepare it into a 10-100 mM stock solution with 100% DMSO (the specific stock solution concentration depends on the properties of the compound); then prepare working solutions of different concentrations using healthy human plasma as the solution (specifically: 0, 0.3, 1, 3, 10, 30, 60, 100, 300 μM), mix thoroughly; incubate at 37°C for 3 min and then perform APTT determination on a machine (model CS-2000I).

[0205] 3. Data processing

[0206] GraphPad Prism software was used for curve fitting and EC150 value was calculated, that is, the effective concentration of the compound corresponding to 50% extension of APTT relative to the blank control group.

[0207] 4. Test results

[0208] The results are shown in the following table. The APTTEC150 of the tested compounds 2, 4, and 14 are all better than that of the compound in comparative example 1.

[0209] Table 3 Determination results of the anticoagulant effect of human plasma in vitro

[0210] sample APTTEC150(μM) sample APTTEC150(μM) Compound 2 0.26 Compound 4 0.24 Compound 14 0.14 Comparative Example 1 1.1

[0211] Test Example 4: Pharmacokinetic study in rats

[0212] 1. Test samples

[0213] Example Compound 2, Compound 14 and Comparative Example 1.

[0214] 2. Preparation method and environmental requirements of the test substance

[0215] The preparation of test substances was carried out on the conventional workbench in the preparation room.

[0216] 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 2, Compound 14 and Comparative Example 1, respectively.

[0217] Preparation of dosing solution for rats: 0.5% CMC-Na was used as solvent. The injection concentration was 3 mg / mL.

[0218] 3. Test operation

[0219] (1) Dosage regimen

[0220] The experiment used 18 SD rats, half male and half female, divided into 6 groups, 3 rats in each group. The dosage of each group was 3 mg / kg.

[0221] (2) Medication and sample collection

[0222] Rats were fasted for 12 hours before administration and had free access to water. The experiment used 18 SD rats, half male and half female, divided into 6 groups, 3 rats in each group. Group 1 was the comparative example 1-♀ group, and the dosage was 3 mg / kg; Group 2 was the comparative example 1-♂ group, and the dosage was 3 mg / kg; Group 3 was the compound 2-♀ group, and the dosage was 3 mg / kg; Group 4 was the compound 2-♂ group, and the dosage was 3 mg / kg; Group 5 was the compound 14-♀ group, and the dosage was 3 mg / kg; Group 6 was the compound 14-♂ group, and the dosage was 3 mg / kg;

[0223] Blank blood was collected before administration, and blood was collected at predetermined time points after administration: 2min, 5min, 10min, 15min, 30min, 45min, 1h, 2h, 3h, 5h, 7h, 24h, 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.

[0224] (3) Animal treatment

[0225] At the end of the experiment, all animals were euthanized according to institutional SOP.

[0226] (4) Instruments

[0227] Liquid chromatography-mass spectrometry analysis system (LC-MS / MS), including Shimadzu LC-20AD series binary pump and SIL-20AC autosampler and AB API-4000Q-Trap mass spectrometer detector (including ESI ion source), chromatographic column: ODS-C18 (4.6×50mm, 3μm).

[0228] (5) Sample processing

[0229] Standard curve sample processing: Prepare a series of working solutions containing different concentrations of comparative example 1 and compound 2 and compound 14. Take 20 μL of the working solution, add 100 μL of blank plasma sample, vortex mix, then add 300 μL of acetonitrile solution containing 40 ng / mL propranolol internal standard, vortex mix, centrifuge at 4°C and 14000g for half an hour, and take the supernatant for LC-MS / MS detection.

[0230] Treatment of rat plasma samples: Take 20 μL of acetonitrile, add 100 μL of plasma sample, vortex mix, then add 300 μL of acetonitrile solution containing 40 ng / mL propranolol internal standard, vortex mix, centrifuge at 4°C and 14000g for half an hour, and take the supernatant for detection.

[0231] (6) Pharmacokinetic analysis

[0232] According to the plasma concentration data of the drug, DAS2.0 software was used to calculate the pharmacokinetic parameters.

[0233] 4. Results

[0234] The experimental results of the pharmacokinetic study of injection administration in rats are shown in the following table. The AUC of Example Compound 2 and Compound 14 are much higher than that of Comparative Example 1, and the half-life is shorter, indicating that Example Compound 2 and Compound 14 have the characteristics of faster onset, stronger efficacy and rapid elimination compared with Comparative Example 1, which reduces the risk of bleeding caused by long-term non-elimination of the drug during use, and the medication process is more controllable.

[0235] Table 4 Comparison of pharmacokinetics in rats

[0236]

[0237] 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 or a pharmaceutically acceptable salt as shown below: 。 2. The compound or pharmaceutically acceptable salt according to claim 1, characterized in that The hydrogen in the structure of the compound or pharmaceutically acceptable salt may be replaced by one or more deuterium.

3. Use of the compound or pharmaceutically acceptable salt according to claim 1 or 2 in the preparation of a medicament for treating and / or preventing diseases associated with FXIa receptor.

4. The use according to claim 3, characterized in that The disease associated with FXIa receptor is selected from cerebrovascular arterial disease or peripheral arterial disease.

5. The use according to claim 3, characterized in that The disease associated with FXIa receptor is selected from transient ischemic attack, ischemic stroke, peripheral arterial occlusion or stent thrombosis.

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