CN116262724A - Novel oxopyridine compound and its preparation method and use - Google Patents

Abstract

The present invention discloses a compound represented by formula (I), its stereoisomer, or a pharmaceutically acceptable salt. The present invention also provides the application of said compound, its stereoisomer or pharmaceutically acceptable salt in the preparation of medicines for the treatment and/or prevention of diseases related to FⅪa receptors, especially in the preparation of treatment and/or prevention of Application of drugs for cerebrovascular arterial disease and/or peripheral arterial disease.

Description

Novel oxopyridine compound and its preparation method and use

technical field

The present invention relates to the field of medicinal chemistry, in particular to oxopyridine compounds or their salts, isomers and preparation methods thereof and their use in the preparation, treatment and/or prevention of diseases related to FⅪa receptors, especially in cerebrovascular and arterial diseases And/or the purposes in the treatment and prevention of peripheral arterial disease etc.

Background technique

Thromboembolism is a disease of humans and animals caused by abnormal blood clots that form in blood vessels during their lifetime. 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 be clinically manifested as myocardial infarction, stroke, deep vein thrombosis (deep vein thrombosis, DVT), pulmonary embolism, atrial fibrillation and cerebral infarction, etc., especially with Embolism and infarction are the main causes of myocardial infarction, cerebral infarction, and pulmonary infarction, ranking first among all causes of death. The world claims nearly 12 million lives every year, which is close to a quarter of the world's total death toll.

Coagulation factor XI (FXI) is a plasma serine proteaseogen necessary to maintain the intrinsic pathway. After activation, it generates activated coagulation factor XIa (FXIa), which plays a key role in the amplification process of the coagulation cascade reaction. In the blood coagulation cascade reaction, thrombin can feed back and activate FXI, and the activated FXI can promote the massive production of thrombin, thereby amplifying the blood coagulation cascade reaction. Therefore, drugs targeting FXI can block the intrinsic pathway and inhibit the amplification of the coagulation cascade, thereby having antithrombotic effects. In recent years, clinical data on human coagulation factor XI (FXI) deficiency or elevated FXI levels associated with thrombotic diseases, as well as animal FXI deficiency or knockout or inhibited antithrombotic experimental studies have shown that compared with direct FXa inhibition Inhibiting FXI may reduce the risk of bleeding, and it is a new target for antithrombotic prevention and treatment.

The reported FXI inhibitors mainly include monoclonal antibodies, antisense oligonucleotides, chemical small molecules, polypeptides or proteins, and polypeptide mimics. At present, milvexian, jointly developed by BMS and Johnson & Johnson, has completed phase II clinical trials, and the results show that it has a small risk of bleeding. The phase I clinical trial of BMS-962122, an intravenous injection of small molecule FXIa inhibitor, has been completed, and the research and development has been suspended. ONO-7684, a small-molecule oral FXIa inhibitor developed by Ono Corporation of Japan, has entered phase I clinical research. Monoclonal antibodies and antisense oligonucleotides need to be administered by injection, and have disadvantages such as expensive, slow onset of action, and may not be easy to control. Chemical small molecules have relatively good oral bioavailability and better patient compliance. . Therefore, the development of safe, effective, specific and highly active small-molecule inhibitors of FXIa may make up for the shortcomings of current clinical anticoagulant and antithrombotic drugs that are prone to bleeding complications and meet the unmet clinical needs.

Contents of the invention

The compound of the present invention is a novel oxopyridine compound, and most of the compounds in the examples show good anticoagulant effect and in vitro affinity to FⅪa.

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

in,

R ¹ is selected from C1-C5 alkyl, wherein: the alkyl can be substituted by a substituent selected from the group consisting of fluorine, cyano, hydroxyl, difluoromethyl, trifluoromethyl, C1-C4 alkoxy, C3 -C6 cycloalkoxy, C3-C6 cycloalkyl, six-membered heterocycloalkyl, phenyl, pyridyl, pyrazolyl, oxazolyl, oxadiazolyl, dihydrooxazolyl or isoxazole group, wherein pyridyl, pyrazolyl, oxazolyl, oxadiazolyl, dihydrooxazolyl and isoxazolyl may be substituted by 1 to 2 substituents independently selected from the group consisting of methyl, ethyl and cyclopropyl;

^R2 is independently selected from C1-C5 alkyl, substituted or unsubstituted aryl, or substituted or unsubstituted amino, the substituents are selected from alkyl, cycloalkyl or halogen;

^R3 is independently selected from hydrogen or halogen;

R ⁴ is independently selected from COR ⁵ or COOH, wherein R ⁵ is selected from C1-C5 alkyl.

Further, in the compound described in any one of the above, its stereoisomer or pharmaceutically acceptable salt, the ^R1 is selected from methyl or ethyl, wherein: methyl or ethyl can be selected from the following Substituent substitution: C1-C4 alkoxy, C3-C6 cycloalkoxy, C3-C6 cycloalkyl, oxygen-containing six-membered cycloalkyl, phenyl, pyridyl, pyrazolyl, oxazolyl , oxadiazolyl, dihydrooxazolyl, isoxazolyl, wherein pyridyl, pyrazolyl, oxazolyl, oxadiazolyl, dihydrooxazolyl and isoxazolyl can be replaced by 1 to 2 Substituents independently selected from the following substituents: methyl, ethyl and cyclopropyl;

And/or ^R2 is independently selected from methyl, ethyl, propyl, phenyl, 3,4-dichlorophenyl, N,N-dimethylamino or cyclopropylamino;

and/or ^R3 are independently selected from hydrogen, fluorine or chlorine;

And/or R ⁴ is independently selected from COR ⁵ , COOH, wherein R ⁵ is selected from methyl or ethyl.

Further, in the compound described in any one of the above, its stereoisomer or pharmaceutically acceptable salt, the ^R1 is selected from methyl or ethyl, wherein: methyl or ethyl can be selected from the following Substituent substitution: methoxy, ethoxy, cyclopropoxy, tert-butoxy, isopropoxy, tetrahydropyranyl, 1,4-dioxanyl, furyl, cyclopropyl, Cyclobutyl, cyclopentyl, cyclohexyl, pyridyl, oxadiazolyl, pyrazolyl, isoxazolyl, methyloxadiazolyl, methylpyrazolyl, methylisoxazolyl, cyclopropane Oxadiazolyl, cyclopropylpyrazolyl, cyclopropylisoxazolyl or phenyl;

And/or ^R2 is independently selected from methyl, phenyl, 3,4-dichlorophenyl, N,N-dimethylamino or cyclopropylamino;

And/or R ⁴ is independently selected from COCH ₃ or COCH ₂ CH ₃ .

Further, in the compound described in any one of the above, its stereoisomer or pharmaceutically acceptable salt, the ^R1 is selected from methyl or ethyl, wherein: methyl or ethyl can be selected from the following Substituent substitution: tetrahydro-2H-pyran-2-yl, methoxy, cyclobutyl, pyridin-4-yl, cyclohexyloxy, 5-methyl-1,3,4-oxadiazole -2-yl, 1-methyl-1H-pyrazol-3-yl, 5-methylisoxazol-3-yl or phenyl;

And/or ^R2 is independently selected from methyl, phenyl, 3,4-dichlorophenyl, N,N-dimethylamino or cyclopropylamino;

And/or R ⁴ is independently selected from COCH ₃ or COCH ₂ CH ₃ .

Further, the compound described in any one of the above, its stereoisomer or pharmaceutically acceptable salt includes the following structure:

Further, hydrogen in the structure of any one of the compounds described above, its stereoisomers or pharmaceutically acceptable salts may be replaced by one or more deuteriums.

In another aspect, the present invention provides a method for preparing the compound described in any one of the above, its stereoisomer or pharmaceutically acceptable salt, comprising the following steps:

Step 1: condensation reaction of starting material a and starting material b to generate intermediate c;

Step 2: intermediate c undergoes a substitution reaction with compound d to generate intermediate e;

Step 3: intermediate e undergoes a hydrolysis reaction under alkaline conditions to generate intermediate f;

Step 4: intermediate f undergoes a condensation reaction with compound g to generate a compound of formula (I);

Wherein R ¹ , R ² , R ³ , R ⁴ are as defined in any one of the above.

In three aspects, the present invention provides the use of any one of the above-mentioned compounds, stereoisomers or pharmaceutically acceptable salts thereof in the preparation of medicines for treating and/or preventing diseases associated with FⅪa receptors.

Further, the above-mentioned diseases related to FⅪa receptor are selected from cerebrovascular arterial disease and/or peripheral arterial disease.

Further, the above-mentioned cerebrovascular arterial diseases include, but are not limited to, transient ischemic attack (TIA), ischemic stroke, or events leading to thrombosis and/or thromboembolic origin of 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 (eg, angioplasty, stent placement or surgery and bypass), and/or stent thrombosis.

Further, the above-mentioned ischemic stroke includes but not limited to cardiogenic stroke, non-cardiogenic stroke, stroke caused by aortic or small artery disease, stroke caused by undetermined cause, cryptogenic stroke, embolic stroke or Embolic stroke of undetermined origin.

Further, the aforementioned cardiogenic stroke includes but not limited to stroke caused by atrial fibrillation; the aforementioned non-cardiogenic stroke includes but not limited to lacunar stroke.

Beneficial effects: Compared with the prior art, the present invention has stronger activity on blood coagulation factor FXIa inhibitors and better pharmacokinetic properties, and has the potential to provide an antithrombotic drug with higher activity and better properties for clinical use.

Detailed ways

The present invention will be described in further detail below in conjunction with the examples and experimental examples. The embodiments of the present invention and the experimental examples are only used to illustrate the technical solutions of the present invention, not to limit the present invention. Equivalent replacements in this field all belong to the protection scope of the present invention.

The compound of the present invention, its stereoisomer or pharmaceutically acceptable salt can be prepared by selecting the synthetic route of the embodiment, and according to the needs of substituents or salt formation, the conventional conditions of the reaction raw materials and the reaction solvent are adjusted, All these can be realized by those skilled in the art on the basis of 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 single or mixed in combination with the reaction solvent and the common knowledge or common means of those skilled in the art unless otherwise specified. Elution solvent.

The structures of the compounds were confirmed by nuclear magnetic resonance ( ¹ H NMR) or liquid chromatography-mass spectrometry (LC-MS).

Liquid-mass spectrometry (LC-MS) is Agilent G6120B (matched with liquid phase Agilent 1260); nuclear magnetic resonance ( ¹ HNMR) is Bruker AVANCE-400 or Bruker AVANCE-800, nuclear magnetic resonance ( ¹ H NMR) shift ( δ ) is given in parts per million (ppm), the measurement solvent is DMSO, the internal standard is tetramethylsilane (TMS), and the chemical shifts are given in units of 10 ⁻⁶ (ppm).

The term "room temperature" in the present invention means that the temperature is between 10°C and 30°C.

Example 1: (S)-4-(2-(4-(2-acetyl-5-chlorophenyl)-5-methoxy-2-oxopyridin-1(2H)-yl)butanamide base)-2-fluoro-N-(methylsulfonyl)benzamide (compound 1)

Step 1: Preparation of (S)-4-(2-bromobutyramide)-2-fluorobenzoic acid methyl ester

Add (R)-2-bromobutyric acid (400mg, 2.39mmol), THF 5ml, methyl 4-aminobenzoate (271mg, 1.6mmol) into a 25ml reaction bottle, stir and cool down to 0°C, add pyridine (475mg, 6mmol) , T3P (50%EA) (2.04g, 3.2mmol) was added dropwise, the dropwise addition was completed, and the reaction was carried out at room temperature for 5h. The reaction of the raw materials was completed, and the reaction was terminated. Add EA, wash twice with acid, wash once with alkali, separate layers, and concentrate the organic phase to dryness to obtain 580 mg of the title product with a yield of 76.3% and a purity of 96.5%.

ESI-MS: m/z=318.0 (M+H) ⁺ .

Step 2: (S)-4-(2-(4-(2-Acetyl-5-chlorophenyl)-5-methoxy-2-oxopyridin-1(2H)-yl)butanylamino ) Preparation of methyl 2-fluorobenzoate

Add (S)-4-(2-bromobutyramide)-2-fluorobenzoic acid methyl ester (550mg, 1.73mmol), isopropanol 5.5ml, acetone 1.1ml, 4-(2-acetyl- 5-Chlorophenyl)-5-methoxypyridin-2(1H)-one (480mg, 1.73mmol), stirred and cooled to 0°C, added tetramethylguanidine (300mg, 2.6mmol) dropwise, the dropwise addition was completed, Reaction at room temperature for 3h. After the reaction was completed, EA was added, washed once with acid and once with alkali, separated into layers, and concentrated to dry organic phase. The concentrate was separated and purified by chromatographic column (MeOH:DCM=2:100), the product was collected and concentrated to dryness to obtain 350 mg of the title product with a yield of 78.6% and a purity of 98.91%.

ESI-MS: m/z=515.1 (M+H) ⁺ .

Step 3: (S)-4-(2-(4-(2-Acetyl-5-chlorophenyl)-5-methoxy-2-oxopyridin-1(2H)-yl)butyrylamide )-2- The preparation of fluorobenzoic acid

Add (S)-4-(2-(4-(2-acetyl-5-chlorophenyl)-5-methoxy-2-oxopyridin-1(2H)-yl)butanamide to 25ml reaction flask )-Methyl 2-fluorobenzoate (340mg, 0.66mmol), methanol 5ml, weigh lithium hydroxide (166mg, 4.0mmol) and dissolve it in 2.5ml of water, add dropwise to the reaction bottle, react at room temperature for 2h, and the reaction of the raw materials is complete. Add EA, wash once with acid, wash once with saturated NaCl, separate layers, and concentrate the organic phase to dryness to obtain 300 mg of the title product with a yield of 90.7% and a purity of 96.06%.

ESI-MS: m/z=501.1 (M+H) ⁺ .

Step 4: (S)-4-(2-(4-(2-Acetyl-5-chlorophenyl)-5-methoxy-2-oxopyridin-1(2H)-yl)butanamide)- Preparation of 2-fluoro-N-(methylsulfonyl)benzamide

Add (S)-4-(2-(4-(2-acetyl-5-chlorophenyl)-5-methoxy-2-oxopyridin-1(2H)-yl)butanamide to 25ml reaction flask Base)-2-fluorobenzoic acid (100mg, 0.2mmol), THF 1.5ml, stirred and dissolved, added CDI (97mg, 0.6mmol), heated to 40°C for 1h. Add methylsulfonamide (38mg, 0.4mmol), DBU (91.2mg, 0.6mmol), react at 40°C for 2h, and the reaction of the raw materials is complete. Add EA, wash once with acid, wash once with saturated NaCl, separate layers, concentrate the organic phase to dryness, the concentrate is separated and purified by chromatography (MeOH:DCM=8:100), collect the product, concentrate to dryness to obtain 90 mg of the title product, the yield 75.9% with a purity of 98.53%.

ESI-MS: m/z=578.1 (M+H) ⁺ .

¹ HNMR (400 MHz, DMSO-d6) δ: 12.10 (s, 1H), 10.78 (s, 1H), 7.89 (d,1H), 7.75 – 7.53 (m, 3H), 7.47 (d, 1H), 7.34 (dd, 1H), 7.28 (s, 1H), 6.42 (s,1H), 5.59 (dd, 1H), 3.55 (s, 3H), 3.00 (s, 3H), 2.13 (ddd, 2H), 1.23 ( s, 3H), 0.89 (t, 3H).

Example 2: (S)-4-(2-(4-(2-acetyl-5-chlorophenyl)-5-methoxy-2-oxopyridin-1(2H)-yl)butanamide base)-N-(3,4-difluorophenyl)sulfonyl)-2-fluorobenzamide (compound 2)

The preparation method was the same as that in Example 1, except that in step 4, methanesulfonamide was replaced by equimolar 3,4-difluorobenzenesulfonamide to obtain the title compound with a yield of 50.3% and a purity of 97.20%.

ESI-MS: m/z = 676.1 (M+H) ⁺ .

¹ HNMR (400 MHz, DMSO-d6) δ: 12.12 (s, 1H), 10.77 (s, 1H), 7.86 (d,1H), 7.83 – 7.75 (m, 3H), 7.75 – 7.64 (m, 2H) , 7.62 (d, 1H), 7.51 – 7.37 (m,3H),, 6.42 (s, 1H), 5.59 (dd, 1H), 3.00 (s, 3H), 2.13 (ddd, 2H), 1.23 (s, 3H), 0.89 (t, 3H).

Example 3: (S)-4-(2-(4-(2-acetyl-5-chlorophenyl)-5-methoxy-2-oxopyridin-1(2H)-yl)butanamide base)-N-(N-cyclopropylsulfonamido)-2-fluorobenzamide (compound 3)

The preparation method was the same as that in Example 1, except that methylsulfonamide in step 4 was replaced by equimolar N-cyclopropylaminosulfonamide to obtain the title compound with a yield of 54.3% and a purity of 96.9%.

ESI-MS: m/z = 619.1 (M+H) ⁺ .

¹ HNMR (400 MHz, DMSO-d6) δ: 12.03 (s, 1H), 9.52 (s, 1H), 7.86 (d,1H), 7.83 – 7.75 (m, 2H), 7.67 (dd, 1H), 7.62 (d, 1H), 7.54 (d, 1H), 7.47(ddd, 2H), 7.05 (s, 1H), 4.82 (td, 1H), 3.81 (s, 3H), 2.66 (dp, 1H), 2.56 ( s,3H), 2.03 – 1.88 (m, 2H), 1.00 (t, 3H), 0.74 – 0.53 (m, 4H).

Example 4: (S)-4-(2-(4-(2-acetyl-5-chlorophenyl)-5-methoxy-2-oxopyridin-1(2H)-yl)butanamide base)-N-(N,N-dimethylsulfonamido)-2-fluorobenzamide (compound 4)

The preparation method was the same as that in Example 1, except that methylsulfonamide in step 4 was replaced by equimolar N,N-dimethylsulfonamide to obtain the title compound with a yield of 76.5% and a purity of 97.87%.

ESI-MS: m/z = 607.1 (M+H) ⁺ .

¹ HNMR (400 MHz, DMSO-d6) δ: 12.10 (s, 1H), 10.78 (s, 1H), 7.89 (d,1H), 7.75 – 7.53 (m, 3H), 7.47 (d, 1H), 7.34 (dd, 1H), 7.28 (s, 1H), 6.42 (s,1H), 5.59 (dd, 1H), 3.81 (s, 3H), 2.70 (s, 6H), 2.56 (s, 3H), 2.03 – 1.88 (m, 2H), 1.00 (t, 3H).

Example 5: (S)-4-(2-(4-(2-acetyl-5-chlorophenyl)-5-methoxy-2-oxopyridin-1(2H)-yl)-3 Preparation of -phenylpropanylamino)-2-fluoro-N-(methylsulfonyl)benzamide (compound 5)

The preparation method is the same as that in Example 1, replacing (R)-2-bromobutyric acid with equimolar (R)-2-bromo-3-phenylpropionic acid in step 1 to obtain the title compound, four-step reaction Yield: 22.1%, purity 97.87%.

ESI-MS: m/z = 640.1 (M+H) ⁺ .

¹ HNMR (400 MHz, DMSO-d6) δ: 10.74 (s, 1H), 7.88 (d, 1H), 7.83 – 7.75(m, 2H), 7.67 (dd, 1H), 7.62 (d, 1H), 7.51 – 7.42 (m, 2H), 7.29 – 7.13 (m,5H), 7.04 (s, 1H), 5.08 (td, 1H), 3.81 (s, 3H), 3.25 (s, 3H), 3.24 – 3.13 (m ,2H), 2.56 (s, 3H).

Example 6: 4-(2S)-2-(4-(2-acetyl-5-chlorophenyl)-5-methoxy-2-oxopyridin-1(2H)-yl)-3- Preparation of (tetrahydro-2H-pyran-2-yl)propionamido)-2-fluoro-N-(methylsulfonyl)benzamide (compound 6)

The preparation method is the same as that in Example 1, replacing (R)-2-bromobutyric acid in step 1 with equimolar (2R)-2-bromo-3-(tetrahydro-2H-pyran-2-yl ) propionic acid to obtain the title compound, the four-step reaction yield: 18.6%, and the purity is 98.23%.

ESI-MS: m/z = 648.1 (M+H) ⁺ .

¹ HNMR (400 MHz, DMSO-d6) δ: 10.35 (s, 1H), 7.86 (d, 1H), 7.83 – 7.75(m, 2H), 7.67 (dd, 1H), 7.62 (d, 1H), 7.51 – 7.42 (m, 2H), 7.05 (s, 1H), 4.86(td, 1H), 3.93 (tt, 1H), 3.81 (s, 3H), 3.74 – 3.57 (m, 2H), 3.25 (s, 3H ), 2.56 (s, 3H), 2.28 – 2.08 (m, 2H), 1.78 – 1.46 (m, 6H).

Example 7: (S)-4-(2-(4-(2-acetyl-5-chlorophenyl)-5-methoxy-2-oxopyridin-1(2H)-yl)-4 Preparation of -methoxybutanylamino)-2-fluoro-N-(methylsulfonyl)benzamide (compound 7)

The preparation method is the same as the preparation method of Example 1, replacing (R)-2-bromobutyric acid with equimolar (R)-2-bromo-4-methoxybutyric acid in step 1 to obtain the title compound, four steps Reaction yield: 24.7%, purity 98.41%.

ESI-MS: m/z = 608.1 (M+H) ⁺ .

¹ HNMR (400 MHz, DMSO-d6) δ: 10.74 (s, 1H),7.81 (d, 1H), 7.79 (dd,1H), 7.70 – 7.59 (m, 3H), 7.51 – 7.42 (m, 2H) , 7.05 (s, 1H), 4.84 (td, 1H), 3.81 (s, 3H), 3.60 – 3.46 (m, 2H), 3.25 (s, 3H), 3.17 (s, 3H), 2.56 (s, 3H ),2.29 – 2.08 (m, 2H).

Example 8: (S)-4-(2-(4-(2-acetyl-5-chlorophenyl)-5-methoxy-2-oxopyridin-1(2H)-yl)-3 Preparation of -cyclobutylpropionylamino)-2-fluoro-N-(methylsulfonyl)benzamide (compound 8)

The preparation method is the same as the preparation method of Example 1, replacing (R)-2-bromobutyric acid with equimolar (R)-2-bromo-4-cyclobutylbutyric acid in step 1 to obtain the title compound, four steps Reaction yield: 16.5%, purity 98.63%.

ESI-MS: m/z = 618.1 (M+H) ⁺ .

¹ HNMR (400 MHz, DMSO-d6) δ: 10.56 (s, 1H),7.86 (d, 1H), 7.79 (dd,1H), 7.70 (d, 1H), 7.67 (dd, 1H), 7.62 (d , 1H), 7.51 – 7.42 (m, 2H), 7.05 (s,1H), 4.86 (td, 1H), 3.81 (s, 3H), 3.25 (s, 3H), 2.56 (s, 3H), 2.04 – 1.79 (m, 3H), 1.76 – 1.56 (m, 4H), 1.53 – 1.37 (m, 2H).

Example 9: (S)-4-(2-(4-(2-acetyl-5-chlorophenyl)-5-methoxy-2-oxopyridin-1(2H)-yl)-3 Preparation of -(pyridin-4-yl)propionamido)-2-fluoro-N-(methylsulfonyl)benzamide (compound 9)

The preparation method is the same as that in Example 1, replacing (R)-2-bromobutyric acid with equimolar (R)-2-bromo-3-(pyridin-4-yl)propionic acid in step 1 to obtain the title Compound, four-step reaction yield: 26.6%, purity 98.92%.

ESI-MS: m/z = 641.1 (M+H) ⁺ .

¹ HNMR (400 MHz, DMSO-d6) δ: 10.44(s, 1H), 8.55 – 8.49 (m, 2H), 7.86(d, 1H), 7.79 (dd, 1H), 7.75 (d, 1H), 7.67 (dd, 1H), 7.62 (d, 1H), 7.51 –7.42 (m, 2H), 7.23 – 7.18 (m, 2H), 7.04 (s, 1H), 5.08 (td, 1H), 3.81 (s, 3H ), 3.47 (dd, 1H), 3.25 (s, 3H), 3.20 (dd, 1H), 2.56 (s, 3H).

Example 10: (S)-4-(2-(4-(2-acetyl-5-chlorophenyl)-5-methoxy-2-oxopyridin-1(2H)-yl)-4 Preparation of -(cyclohexyloxy)butanylamino)-N-(methylsulfonyl)benzamide (compound 10)

The preparation method is the same as that in Example 1, replacing (R)-2-bromobutyric acid with equimolar (R)-2-bromo-4-(cyclohexyloxy)butanoic acid in step 1, 4-amino The title compound was obtained by replacing methyl 2-fluorobenzoate with equimolar methyl 4-amino-benzoate. The yield of the four-step reaction was 14.8%, and the purity was 98.22%.

ESI-MS: m/z = 658.2 (M+H) ⁺ .

¹ HNMR (400 MHz, DMSO-d6) δ: 10.61(s, 1H), 7.95 – 7.88 (m, 2H), 7.86(d, 1H), 7.76 – 7.68 (m, 2H), 7.63 (dd, 2H) , 7.45 (dd, 1H), 7.05 (s, 1H), 4.83 (td, 1H), 3.84 – 3.72 (m, 4H), 3.59 (t, 2H), 3.25 (s, 3H), 2.56 (s, 3H ),2.24 – 2.04 (m, 2H), 1.81 – 1.67 (m, 2H), 1.67 – 1.59 (m, 1H), 1.63 – 1.58(m, 1H), 1.62 – 1.47 (m, 2H), 1.51 – 1.37 (m, 1H), 1.42 – 1.32 (m, 1H).

Example 11: (S)-4-(2-(4-(2-acetyl-5-chlorophenyl)-5-methoxy-2-oxopyridin-1(2H)yl)-3- Preparation of (5-methyl-1,3,4-oxadiazol-2-yl)propionamido)-2-fluoro-N-(methylsulfonyl)benzamide (compound 11)

The preparation method is the same as that in Example 1, replacing (R)-2-bromobutyric acid in step 1 with equimolar (R)-2-bromo-3-(5-methyl-1,3,4- Oxadiazol-2-yl) propionic acid to obtain the title compound, the four-step reaction yield: 16.7%, the purity is 97.82%.

ESI-MS: m/z = 646.1 (M+H) ⁺ .

¹ HNMR (400 MHz, DMSO-d6) δ: 10.44(s, 1H),7.86 (d, 1H), 7.79 (dd, 1H),7.70 – 7.64 (m, 2H), 7.62 (d, 1H), 7.51 – 7.42 (m, 2H), 6.94 (s, 1H), 5.17(td, 1H), 3.81 (s, 3H), 3.35 (dd, 2H), 3.25 (s, 3H), 2.56 (s, 3H), 2.52 (s,3H).

Example 12: (S)-4-(2-(4-(2-acetyl-5-chlorophenyl)-5-methoxy-2-oxopyridin-1(2H)-yl)-3 Preparation of -(1-methyl-1H-pyrazol-3-yl)propionamido)-2-fluoro-N-(methylsulfonyl)benzamide (compound 12)

The preparation method is the same as that in Example 1, replacing (R)-2-bromobutyric acid in step 1 with equimolar (R)-2-bromo-3-(1-methyl-1H-pyrazole-3 -yl) propionic acid to obtain the title compound, the four-step reaction yield: 16.7%, and the purity is 97.82%.

ESI-MS: m/z = 644.1 (M+H) ⁺ .

¹ HNMR (400 MHz, DMSO-d6) δ: 10.56(s, 1H), 7.86 (d, 1H), 7.79 (dd, 1H), 7.75 (d, 1H), 7.67 (dd, 1H), 7.62 (d , 1H), 7.51 – 7.42 (m, 2H), 7.39 (dt,1H), 6.96 (s, 1H), 6.06 (d, 1H), 5.09 (td, 1H), 3.87 – 3.79 (m, 5H), 3.25 (s,3H), 3.14 – 3.00 (m, 2H), 2.56 (s, 3H).

Example 13: (S)-4-(2-(4-(5-chloro-2-propionylphenyl)-5-methoxy-2-oxopyridin-1(2H)-yl)-3 Preparation of -(5-methylisoxazol-3-yl)propionamido)-2-fluoro-N-(methylsulfonyl)benzamide (compound 13)

The preparation method is the same as that in Example 1, replacing (R)-2-bromobutyric acid in step 1 with equimolar (R)-2-bromo-3-(5-methylisoxazol-3-yl ) propionic acid, 4-(2-acetyl-5-chlorophenyl)-5-methoxypyridin-2(1H)-one in step 2 was replaced by equimolar 4-(5-chloro-2-propane Acylphenyl)-5-methoxypyridin-2(1H)-one, the title compound was obtained, the yield of the four-step reaction was 13.7%, and the purity was 97.23%.

ESI-MS: m/z = 659.1 (M+H) ⁺ .

¹ HNMR (400 MHz, DMSO-d6) δ: 10.16(s, 1H), 7.85 (d, 1H), 7.79 (dd, 1H), 7.75 (d, 1H), 7.67 (dd, 1H), 7.57 (d , 1H), 7.51 – 7.41 (m, 2H), 6.96 (s, 1H),6.21 (d, 1H), 5.12 (td, 1H), 3.81 (s, 3H), 3.25 (s, 3H), 3.16 – 2.97 (m, 4H), 2.44 (s, 3H), 1.17 (t, 3H).

Example 14: (S)-4-chloro-2-(5-methoxy-1-(1-(4-((methylsulfonyl)carbamoyl)phenyl)-1-oxo-3- Preparation of phenylpropanamide-2-yl)-2-oxo-1,2-dihydropyridin-4-yl)benzoic acid (compound 14)

The preparation method is the same as that in Example 1, replacing (R)-2-bromobutyric acid in step 1 with equimolar (R)-2-bromo-3-phenylpropionic acid, 4-amino-2-fluoro Methyl benzoate was replaced by an equimolar amount of 4-amino-methylbenzoate, 4-(2-acetyl-5-chlorophenyl)-5-methoxypyridin-2(1H)-one in step 2 For equimolar tert-butyl 4-chloro-2-(5-methoxy-2-oxo-1,2-dihydropyridin-4-yl)benzoate, (S)-4-chloro-2 -(5-Methoxy-1-(1-((4-((methylsulfonyl)carbamoyl)phenyl)amino)-1-oxo-3-phenylpropanamide-2-yl)- 2-Oxo-1,2-dihydropyridin-4-yl) tert-butyl benzoate, followed by hydrolysis reaction under 2M HCl/dioxane solution to give the title compound, the yield of the five-step reaction: 11.8% , with a purity of 98.62%.

ESI-MS: m/z = 624.1 (M+H) ⁺ .

¹ HNMR (400 MHz, DMSO-d6) δ: 13.11 (s, 1H), 10.74 (s, 1H), δ: 7.95 –7.86 (m, 3H), 7.78 – 7.68 (m, 3H), 7.65 (d, 1H), 7.35 (dd, 1H), 7.29 – 7.18(m, 3H), 7.23 – 7.13 (m, 2H), 7.01 (s, 1H), 5.09 (td, 1H), 3.81 (s, 3H), 3.25 (s, 3H), 3.24 – 3.13 (m, 2H).

Comparative Example 1: (S)-4-(2-(4-(2-acetyl-5-chlorophenyl)-5-methoxy-2-oxopyridin-1(2H)-yl)-3 -Phenyl

Preparation of propionylamino) benzoic acid

Synthesized according to the method described in patent CN113929618A, purity: 98.5%.

ESI-MS: m/z = 575.2 (M+H) ⁺ .

¹ H NMR (400 MHz, DMSO-d6) δ: 10 .81(s ,1H) ,7 .92(s ,1H) ,7 .90(s ,1H) , 7 .83-

7.81(d,1H) ,7.74(s,1H) ,7.72(s,1H) ,7.62-7.59(dd,1H) ,7.43(s,1H)7 . 38(s ,1H) ,7 .30-7 .25(m,4H) ,7 .21-7 .17(m ,1H) ,8 .31(s ,1H) ,6 .05-6 .01( m ,1H) ,3 .54(s ,3H) ,3 .49-3 .44(m ,2H) ,2 .37(s ,3H).

Test Example 1: Determination of blood coagulation factor FXIa inhibitor activity (50nM inhibition rate of blood coagulation factor FXIa)

1. Test sample

Example Compounds 1 to 14 and Comparative Example 1.

⒉Test steps

1) Prepare the experimental buffer (50mM HEPES, 5mM KCl, 145mM NaCl, 1mg/ml PEG8000, pH7.4), and equilibrate to room temperature.

2) Prepare 10X compound working solution.

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

4) Add 20 μL of the 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.

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

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

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

8) After the incubation is completed, use EnVision to read the absorbance value of OD405nm and collect the data.

⒊Data analysis

1) Z' factor = 1-3*(SD _Max +SD _Min )/(Mean _Max -Mean _Min );

2) CV _Max = (SD _Max /Mean _Max )*100%;

3) CVMin = (SD _Min /Mean _Min )*100%;

4) S/B = Signal/Background;

5) blank control: 0.1% DMSO; positive control: comparative example 1;

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

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

⒋Experimental results

As shown in the following table: at 50nM, the inhibitory effect of the compounds of the present invention on FXIa is stronger than that of Comparative Example 1.

Test Example 2: Determination of blood coagulation factor FXIa inhibitor activity (IC ₅₀ )

Utilize the in vitro enzymatic test to determine the inhibitory effect of compound 2, 5, 6, 9, 10, 11, 12, 13 and the compound of comparative example 1 on human blood coagulation factor FXIa, set 5 concentrations, namely: 200nM, 40nM, 8nM, 1.6nM, 0.32nM, detection IC50 value.

Test reagents and operating methods are as described in Test Example 1.

The test results are shown in the following table: Compounds 2, 5, 6, 9, 10, 11, 12, and 13 of the present invention have stronger inhibitory effects on FXIa than the compound of Comparative Example 1.

Test Example 3: Pharmacokinetic Study in Rats

1. Test sample

Example compound 2 and comparative example 1.

2. Test substance preparation method and environmental requirements

The preparation of the test substances was carried out on the regular workbench in the preparation room.

Stock solution preparation according to the proposed method: prepared according to the dispensing regulations, using methanol as the solvent to prepare 1.00 mg/mL stock solutions of Example Compound 2 and Comparative Example 1 respectively.

Rat administration solution preparation: use 0.5% CMC-Na as solvent preparation. The oral administration concentration is 3 mg/mL.

⒊Test operation

(1) Dosing regimen

In the experiment, 12 SD rats, half male and half male, were divided into 4 groups with 3 rats in each group. The dosage of each group was 3 mg/kg.

(2) Drug administration and sample collection

Rats were fasted for 12 h before administration and had free access to water. In the experiment, 12 SD rats, half male and half male, were divided into 4 groups with 3 rats in each group. Group 1 is the comparative example 1-♀ group, the dosage is 3 mg/kg; Group 2 is the comparative example 1-♂ group, the dosage is 3 mg/kg; Group 3 is the compound 2-♀ group, the dosage is 3mg /kg; 4 groups are compound 2-♂ group, administration dose 3mg/kg;

Collect blank blood before administration, and collect blood at predetermined time points after administration: 5min, 15min, 30min, 45min, 1h, 2h, 3h, 5h, 7h, 24h, about 0.5mL of blood is collected, placed in an EDTA-K2 tube, and centrifuged Plasma was stored at -80°C.

(3) Animal disposal

After the experiment, all animals were euthanized according to institutional SOP.

(4) Instruments

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

(5) Sample processing

Standard curve sample processing: Prepare a series of working solutions containing different concentrations of Comparative Example 1 and Compound 2. Take 20 μL working solution, add 100 μL blank plasma sample, vortex mix, then add 300 μL acetonitrile solution containing 40 ng/mL propranolol internal standard, vortex mix, centrifuge at 4°C, 14000g for half an hour, take The supernatant was subjected to LC-MS/MS detection.

Rat plasma sample processing: take 20 μL acetonitrile, add 100 μL plasma sample, vortex mix, then add 300 μL acetonitrile solution containing 40 ng/mL propranolol internal standard, vortex mix, centrifuge at 4°C, 14000g for half Hours, the supernatant was taken for detection.

(6) Pharmacokinetic analysis

According to the plasma concentration data of the drug, the DAS 2.0 software is used to calculate the pharmacokinetic parameters, providing parameters such as AUC _last , C _max , T _max , CLz, t _1/2 .

⒋ test results

The results of oral pharmacokinetic characteristics of compound 2 and comparative example 1 in rats are shown in the table below. Compound 2 has a pharmacokinetic characteristic superior to that of comparative example 1, indicating that the oral absorption of compound 2 in rats is better than that of comparative example 1. In particular, the clinical results of Comparative Example 1 show that multiple daily doses are required and the dosage is large, while the half-life of Compound 2 of the present invention is longer than that of Comparative Example 1, which can reduce the number of doses.

The above-mentioned embodiment is only one of the preferred implementation modes of the present invention, and should not be used to limit the scope of protection of the present invention, but any modification or embellishment without substantive significance made on the main design idea and spirit of the present invention shall be solved by If the technical problems are still consistent with the present invention, all should be included in the protection scope of the present invention.

Claims (10)

1. A compound represented by formula (I), its stereoisomer or pharmaceutically acceptable salt:

in, R ¹ is selected from C1-C5 alkyl, wherein: the alkyl can be substituted by a substituent selected from the group consisting of fluorine, cyano, hydroxyl, difluoromethyl, trifluoromethyl, C1-C4 alkoxy, C3 -C6 cycloalkoxy, C3-C6 cycloalkyl, six-membered heterocycloalkyl, phenyl, pyridyl, pyrazolyl, oxazolyl, oxadiazolyl, dihydrooxazolyl or isoxazole group, wherein pyridyl, pyrazolyl, oxazolyl, oxadiazolyl, dihydrooxazolyl and isoxazolyl may be substituted by 1 to 2 substituents independently selected from the group consisting of methyl, ethyl and cyclopropyl; ^R2 is independently selected from C1-C5 alkyl, substituted or unsubstituted aryl, or substituted or unsubstituted amino, the substituents are selected from alkyl, cycloalkyl or halogen; ^R3 is independently selected from hydrogen or halogen; R ⁴ is independently selected from COR ⁵ or COOH, wherein R ⁵ is selected from C1-C5 alkyl. 2. The compound according to claim 1, its stereoisomer or pharmaceutically acceptable salt, characterized in that, R ¹ is selected from methyl or ethyl, wherein: methyl or ethyl can be substituted by substituents selected from: C1-C4 alkoxy, C3-C6 cycloalkoxy, C3-C6 cycloalkyl , oxygen-containing six-membered cycloalkyl, phenyl, pyridyl, pyrazolyl, oxazolyl, oxadiazolyl, dihydrooxazolyl, isoxazolyl, among which pyridyl, pyrazolyl, oxazolyl , oxadiazolyl, dihydrooxazolyl and isoxazolyl may be substituted by 1 to 2 substituents independently selected from the group consisting of methyl, ethyl and cyclopropyl; And/or ^R2 is independently selected from methyl, ethyl, propyl, phenyl, 3,4-dichlorophenyl, N,N-dimethylamino or cyclopropylamino; and/or ^R3 are independently selected from hydrogen, fluorine or chlorine; And/or R ⁴ is independently selected from COR ⁵ , COOH, wherein R ⁵ is selected from methyl or ethyl. 3. The compound according to claim 1, its stereoisomer or pharmaceutically acceptable salt, characterized in that, ^R is selected from methyl or ethyl, wherein: methyl or ethyl can be substituted by a substituent selected from: methoxy, ethoxy, cyclopropoxy, tert-butoxy, isopropoxy, Tetrahydropyranyl, furyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, pyridyl, oxadiazolyl, pyrazolyl, isoxazolyl, methyloxadiazolyl, methyl Pyrazolyl, methylisoxazolyl, cyclopropyloxadiazolyl, cyclopropylpyrazolyl, cyclopropylisoxazolyl or phenyl; And/or ^R2 is independently selected from methyl, phenyl, 3,4-dichlorophenyl, N,N-dimethylamino or cyclopropylamino; And/or R ⁴ is independently selected from COCH ₃ or COCH ₂ CH ₃ . 4. The compound according to claim 1, its stereoisomer or pharmaceutically acceptable salt, characterized in that, R ¹ is selected from methyl or ethyl, wherein: methyl or ethyl may be substituted by a substituent selected from tetrahydro-2H-pyran-2-yl, methoxy, cyclobutyl, pyridine-4 -yl, cyclohexyloxy, 5-methyl-1,3,4-oxadiazol-2-yl, 1-methyl-1H-pyrazol-3-yl, 5-methylisoxazole-3 -yl or phenyl; And/or ^R2 is independently selected from methyl, phenyl, 3,4-dichlorophenyl, N,N-dimethylamino or cyclopropylamino; And/or R ⁴ is independently selected from COCH ₃ or COCH ₂ CH ₃ . 5. The compound, its stereoisomer or pharmaceutically acceptable salt as claimed in any one of claims 1 to 4, wherein the compound is selected from the following structures:

. 6. The compound, its stereoisomer or pharmaceutically acceptable salt as claimed in any one of claims 1 to 5, wherein the compound, its stereoisomer or pharmaceutically acceptable salt Hydrogens in the structure may be replaced by one or more deuteriums. 7. the preparation method of the compound described in any one of claims 1～5, its stereoisomer or pharmaceutically acceptable salt, it is characterized in that, described method comprises the following steps:

Step 1: condensation reaction of starting material a and starting material b to generate intermediate c; Step 2: intermediate c undergoes a substitution reaction with compound d to generate intermediate e; Step 3: intermediate e undergoes a hydrolysis reaction under alkaline conditions to generate intermediate f; Step 4: intermediate f undergoes a condensation reaction with compound g to generate a compound of formula (I); wherein R ¹ , R ² , R ³ , R ⁴ are as defined in claim 1. 8. Application of the compound according to any one of claims 1 to 6, its stereoisomers or pharmaceutically acceptable salts in the preparation of medicines for the treatment and/or prevention of diseases associated with FⅪa receptors. 9. The application according to claim 8, characterized in that the disease associated with the FXIA receptor is selected from cerebrovascular arterial disease and/or peripheral arterial disease. 10. Use as claimed in claim 8, wherein the disease associated with the FXIA receptor is selected from the group consisting of transient ischemic attack (TIA), ischemic stroke, including cardiogenic stroke, for example due to atrial fibrillation stroke caused by noncardiac origin, such as lacunar stroke, stroke due to disease of a large or small artery, or stroke of undetermined origin, cryptogenic stroke, embolic stroke, embolic stroke of undetermined origin, or events of thrombotic and/or thromboembolic origin leading to stroke or TIA, and/or conditions of peripheral arteries leading to peripheral arterial disease, including peripheral arterial occlusion, acute limb ischemia, amputation, interventions (e.g., angioplasty, stent reocclusion and restenosis after implantation or surgery and bypass), and/or stent thrombosis.