Oxopyrimidine compounds, preparation methods and uses thereof

Abstract

The present invention discloses a compound represented by formula (I), its stereoisomers, or pharmaceutically acceptable salts. The present invention also provides the use of the compound, its stereoisomers or pharmaceutically acceptable salts in the preparation of drugs for treating and / or preventing diseases related to the FXIa receptor, especially in the preparation of drugs for treating and / or preventing cerebrovascular arterial diseases and / or peripheral arterial diseases, with significant pharmacodynamic effects.

Description

Technical Field

[0001] The present invention relates to the field of medicinal chemistry, and particularly to oxopyridine compounds or their salts, isomers, and their preparation methods, as well as their use in the preparation of drugs for treating and / or preventing diseases related to the FXIa receptor, especially their use in the preparation of drugs for treating cerebrovascular arterial diseases and / or peripheral arterial diseases, etc. Background Art

[0002] Thromboembolic diseases are diseases caused by abnormal blood clots formed in blood vessels during the survival period of humans and animals. Clinically, they may manifest as myocardial infarction, stroke, deep vein thrombosis (DVT), pulmonary embolism, atrial fibrillation, and cerebral infarction, etc., claiming the lives of tens of millions of people globally every year. Coagulation factor XI (FXI) is a plasma serine protease zymogen 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. Therefore, drugs targeting the FXIa target can block the intrinsic pathway and inhibit the amplification of the coagulation cascade reaction, thus having an antithrombotic effect.

[0003] The reported FXIa inhibitors mainly include monoclonal antibodies, antisense oligonucleotides, small chemical molecules, polypeptides or proteins, and polypeptide mimetics, etc. Currently, milvexian jointly developed by BMS and Johnson & Johnson has completed clinical phase II trials, and the results show a relatively low bleeding risk. The clinical phase I trial of the intravenous small molecule FXIa inhibitor BMS-962122 of BMS has been completed, and the R & D has been suspended. The small molecule oral FXIa inhibitor ONO-7684 developed by Ono Pharmaceutical Co., Ltd. of Japan has entered clinical phase I research. BAY-2433334 developed by Bayer has completed clinical phase II trials and has become the most promising small molecule FXIa inhibitor currently. Monoclonal antibodies and antisense oligonucleotides need to be administered by injection, and have the disadvantages of high price, slow onset, and possible difficulty in control. Small chemical molecules have the advantages of relatively good oral bioavailability and better patient compliance.

[0004] Therefore, the research and development of new small molecule FXIa inhibitors with high safety, good specificity, and strong activity may make up for the deficiencies of current clinical anticoagulant and antithrombotic drugs that are prone to bleeding complications and meet the unmet clinical needs. Summary of the Invention

[0005] The compounds of the present invention are a novel type of oxopyridine compounds, showing good anticoagulant effects and in vitro and in vivo affinities for FXIa.

[0006] On the one hand, the present invention provides a compound represented by formula (I), its stereoisomers, or a pharmaceutically acceptable salt:

[0007]

[0008] Further, the pharmaceutically acceptable salt is a metal salt.

[0009] Further, the metal salt is selected from sodium salt, potassium salt, calcium salt, lithium salt, and magnesium salt.

[0010] On the other hand, the present invention provides a pharmaceutical composition of the above compound, its stereoisomer or pharmaceutically acceptable salt, and the composition further contains a pharmaceutically acceptable carrier and / or excipient.

[0011] On another aspect, the present invention provides a preparation method of the above compound, its stereoisomer or pharmaceutically acceptable salt, including the following route:

[0012]

[0013] On the other hand, the present invention also provides the use of any one of the above compounds, its stereoisomers or pharmaceutically acceptable salts or their compositions in the preparation of a drug for treating and / or preventing diseases related to the FⅪa receptor.

[0014] Further, the diseases related to the FⅪa receptor are selected from thrombosis or thromboembolism-related disorders.

[0015] Further, the diseases related to the FⅪa receptor are selected from cerebrovascular arterial diseases and / or peripheral arterial diseases.

[0016] Further, the above cerebrovascular arterial diseases include but are not limited to transient ischemic attack (TIA), ischemic stroke or events originating from thrombosis and / or thromboembolism leading to stroke or TIA; the above peripheral arterial diseases include but are not limited to peripheral arterial occlusion, acute limb ischemia, amputation, re-occlusion and restenosis after intervention (such as angioplasty, stent implantation or surgery and bypass), and / or stent thrombosis.

[0017] Further, the above ischemic stroke includes but is not limited to cardiogenic stroke, non-cardiogenic stroke, stroke caused by large artery or small artery diseases, stroke caused by undetermined reasons, cryptogenic stroke, embolic stroke or embolic stroke of undetermined origin.

[0018] Further, the above cardiogenic stroke includes but is not limited to stroke caused by atrial fibrillation; the above non-cardiogenic stroke includes but is not limited to lacunar stroke.

[0019] Beneficial effects: Compared with the prior art, the present invention has good FXIa inhibitory effect. In the rabbit carotid artery thrombosis model induced by FeCl2, the weight of the arterial thrombus in the carotid artery of the rabbits in the group of Example Compound 1 is significantly reduced compared with that in the group of Comparative Example 1, which is statistically significant, proving that the in vitro and in vivo pharmacodynamic effects of the compounds of the present invention are remarkable. Detailed implementation manners

[0020] The present invention will be further described in detail below in conjunction with examples and experimental examples. The examples and experimental examples of the present invention are only used to illustrate the technical solutions of the present invention and do not limit the present invention. Any equivalent substitution in the art made in accordance with the content disclosed in the present invention belongs to the protection scope of the present invention.

[0021] The structure of the compound is determined by nuclear magnetic resonance ( 1 H NMR) or liquid chromatography - mass spectrometry (LC - MS).

[0022] The liquid chromatography - mass spectrometer (LC - MS) is Agilent G6120B (used in conjunction with the liquid phase Agilent1260); the nuclear magnetic resonance spectrometer ( 1 HNMR) is Bruker AVANCE - 400 or Bruker AVANCE - 800. The nuclear magnetic resonance ( 1 HNMR) chemical shift (δ) is given in parts per million (ppm) units, with the internal standard being tetramethylsilane (TMS), and the chemical shift is given in units of 10 -6 (ppm).

[0023] The term "room temperature" in the present invention refers to a temperature between 10 and 30 °C.

[0024] Example 1: Preparation of (S)-4-(2-(4-(5 - chloro - 2-(1H - tetrazol - 1 - yl)phenyl)-5-(methoxy - d3)-2 - oxopyrimidin - 1(2H)-yl)butanamido)-2 - fluoro - N-(methyl - d3)benzamide (Compound 1):

[0025]

[0026] Step 1: Preparation of intermediate b

[0027] Take compound a (2 g, 15.8 mmol), dissolve it in 40 ml of DMF, add cesium carbonate (10.3 g, 31.6 mmol), cool to 0 °C, and dropwise add methyl iodide-d (3.5 g, 24.1 mmol). It takes about 30 minutes to add it all. After adding, stir the reaction at room temperature for 4 hours. Add EA and water for extraction, wash with water, wash with saturated brine, dry over anhydrous sodium sulfate, filter, evaporate the solvent, and purify by column chromatography to obtain 2.1 g of intermediate b. Yield: 92.84%, HPLC purity: 98.31%.

[0028] ESI-MS: m / z = 144.1 (M+H) + 。

[0029] Step 2: Preparation of intermediate c

[0030] Take diisopropylamine (1.7 g, 16.8 mmol) and dissolve it in 20 ml of THF. Cool to below -60 °C, and dropwise add 6.4 ml of a 2.5 M n-butyllithium hexane solution. It takes about 1 hour to add it all. After adding, stir the reaction at -60 °C for 15 minutes. Dropwise add a 5 ml THF solution of compound b (2 g, 14.0 mmol), which takes about 1 hour. After adding, stir the reaction at -60 °C for 2 hours. Dropwise add triisopropyl borate (2.9 g, 15.4 mmol), which takes 30 minutes. After adding, slowly warm up to room temperature (20 °C) and stir the reaction for 30 minutes. Dropwise add a mixture of 3 g of acetic acid and 15 g of water to terminate the reaction. After adding, stir at room temperature for 30 minutes. Distill off the organic solvent, add a little water, stir at room temperature for 15 minutes, filter, wash the filter cake with water, and dry it under vacuum at 70 °C to obtain 2.03 g of solid. Yield 77.54%, HPLC purity: 98.54%.

[0031] ESI-MS: m / z = 188.1 (M+H) + 。

[0032] Step 3: Preparation of intermediate e

[0033] Take compound d (2.59 g, 10.0 mmol), and suspend Pd(amphos)Cl2 (107.5 mg, 0.15 mmol) in 25 ml of tert-amyl alcohol. Heat to 85 °C, and dropwise add a mixed solution of compound c (2.22 g, 11.9 mmol), sodium carbonate (3.2 g, 30.2 mmol), and 25 ml of water, which takes about 1 hour. After adding, react at 85 °C for 1 hour. Cool to room temperature, add EA / water for extraction treatment, separate the aqueous layer, wash the organic layer successively with water and saturated brine, dry over anhydrous sodium sulfate, filter, and evaporate the solvent. Purify by column chromatography to obtain 2.58 g of intermediate e. Yield 80.18%, HPLC purity: 98.63%.

[0034] ESI-MS: m / z = 322.1 (M+H) + 。

[0035] Step 4: Preparation of Intermediate f

[0036] Compound e (1.87 g, 5.80 mmol), anhydrous lithium chloride (1.3 g, 30.7 mmol), p-toluenesulfonic acid monohydrate (2.2 g, 11.6 mmol) and 20 ml of isopropanol were mixed, heated under reflux for 16 hours. Cooled to room temperature, half of the solvent was distilled off, 30 ml of water was added, stirred at room temperature for 15 minutes, filtered, the filter cake was washed with water, and dried in vacuo at 70 °C to obtain 1.59 g of solid. Yield: 89.09%, HPLC purity: 96.37%.

[0037] ESI-MS: m / z = 308.1 (M+H) + 。

[0038] Step 5: Preparation of Intermediate h

[0039] A 25 ml reaction flask was charged with compound f (523 mg, 1.70 mmol), tetramethylguanidine (681 mg, 5.91 mmol), 6 ml of isopropanol, 1.5 ml of acetone, stirred at room temperature for 15 minutes, compound g (684 mg, 1.9 mmol) was added, and stirred at room temperature overnight. The next day, the reaction was terminated by adding water, extracted with EA, the aqueous layer was separated, and the organic layer was washed successively with saturated ammonium chloride, water, saturated brine, dried over anhydrous sodium sulfate, filtered, the solvent was evaporated, purified by column chromatography, the product was collected, and 857 mg of pure product was obtained. Yield 85.87%, purity 98.69%.

[0040] ESI-MS: m / z = 587.1 (M+H) + 。

[0041] Step 6: Preparation of Intermediate j

[0042] A 25 ml reaction flask was charged with compound h (587 mg, 1 mmol), 8 ml of methanol, stirred to dissolve, and cooled to 0 °C. Lithium hydroxide monohydrate (84 mg, 2 mmol) was weighed and dissolved in 4 ml of water, added dropwise to the reaction flask. After addition, the reaction was carried out at room temperature for 2 h. The reaction was terminated by adding water, the pH was adjusted to weakly acidic with 5% citric acid, extracted with EA, the aqueous layer was separated, and the organic layer was washed successively with water, saturated NaCl, dried over anhydrous sodium sulfate, filtered, the solvent was evaporated, and column chromatography was carried out to collect 419 mg of intermediate j, with a yield of 78.92% and a purity of 98.51%.

[0043] ESI-MS: m / z = 531.1 (M+H) + 。

[0044] Step 7: Preparation of (S)-4-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-5-(methoxy-d3)-2-oxopyrimidin-1(2H)-yl)butanamido)-2-fluoro-N-(methyl-d3)benzamide (Compound 1)

[0045] Add intermediate j (0.3 g, 0.57 mmol), 3 ml of DMF, deuterated methylamine hydrochloride (48.2 mg, 0.68 mmol), EDCI (218.7 mg, 1.14 mmol), and HOBT (154 mg, 1.14 mmol) to a 50 ml reaction flask. Cool the mixture to 0 °C and add DIPEA (295 mg, 2.28 mmol) dropwise. React at room temperature for 12 h until the raw materials are completely reacted. Add the reaction solution to cold water, extract twice with ethyl acetate, wash twice with water, wash with saturated brine, dry over anhydrous sodium sulfate, evaporate the solvent, and purify by silica gel column chromatography to obtain 225 mg of Compound 1 with a yield of 72.17% and a purity of 98.49%.

[0046] ESI-MS: m / z = 547.2 (M+H) + 。

[0047] 1 1H NMR (400 MHz, DMSO-d6) δ: 10.70 (s, 1H), 9.09 (s, 1H), 8.01 (s, 1H), 7.83–7.66 (m, 3H), 7.71–7.60 (m, 2H), 7.35 (m, 1H), 7.10 (m, 1H), 6.95 (s, 1H), 2.09 (m, 2H), 0.77 (t, J = 7.2 Hz, 3H).

[0048] 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)butanamido)-2-fluorobenzamide

[0049]

[0050] Synthesized according to the method described in Patent CN108026072B, purity: 98.5%.

[0051] ESI-MS: m / z = 593.1 (M+H) + 。

[0052] 11H 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).

[0053] Test Example 1: Inhibitory Effect on Coagulation Factor FXIa

[0054] 1. Test Samples

[0055] Compound 1 of the Example and Comparative Example 1.

[0056] 2. Test Procedures

[0057] 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.

[0058] 2) Prepare a 10X compound working solution.

[0059] 3) Prepare a 0.8 nM Human FXIa working solution (2X) and mix well for later use.

[0060] 4) Add 20 μL of the FXIa working solution in step 3) to all the experimental wells of a 384-well plate (Coring, 3702), centrifuge at 200 g, RT for 10 s.

[0061] 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.

[0062] 6) Prepare a 750 μM S-2366 working solution (2.5X) and mix well for later use.

[0063] 7) Add 16 μL of the S-2366 working solution in step 6) to all the experimental wells of the 384-well plate, centrifuge at 200 g, RT for 10 s, and then incubate the working plate at 37 °C for 45 min.

[0064] 8) After incubation, use EnVision to read the absorbance at OD405 nm and collect the data.

[0065] Set 5 concentrations, namely: 200 nM, 40 nM, 8 nM, 1.6 nM, 0.32 nM, and detect the IC 50 value.

[0066] 3. Data analysis

[0067] 1) Z’ factor = 1 - 3*(SD Max + SD Min ) / (Mean Max - Mean Min );

[0068] 2) CV Max = (SD Max / Mean Max ) * 100%;

[0069] 3) CVMin = (SD Min / Mean Min ) * 100%;

[0070] 4) S / B = Singal / Background;

[0071] 5) Blank control: 0.1% DMSO; Positive control: Comparative Example 1;

[0072] 6) Calculation formula of IC 50 : Y = Bottom + (Top - Bottom) / (1 + 10^((LogIC 50 - X) * HillSlope)).

[0073] X: Log value of compound concentration; Y: Inhibition%.

[0074] 4. Test results

[0075] The test results are shown in the following table. The results show that under the condition of the same molar concentration, the compound of the present invention has an in vitro inhibitory activity against FXIa equivalent to that of the compound of Comparative Example 1.

[0076] Table 1 Determination of the activity of coagulation factor FXIa inhibitor (IC50)

[0077] Sample <![CDATA[IC 50 (nM)]]> Sample <![CDATA[IC 50 (nM)]]> Compound 1 0.4 Comparative Example 1 7.6

[0078] Test Example 2: In vivo pharmacodynamic evaluation of rabbit arteriovenous shunt model

[0079] 1. Test samples

[0080] Compound 1 of the example and Comparative Example 1.

[0081] 2. Test method

[0082] Select New Zealand white rabbits, all male, 30 rabbits, 2.5 - 3.0 kg. Divide them into 3 groups, 10 rabbits / group. They are the model group, the Comparative Example 1 group and the Compound 1 group respectively.

[0083] Rabbits in the control group 1 and the compound 1 group were each intravenously injected via the femoral vein with the compounds shown in control 1 and compound 1 at a dose of 6 mg / kg.

[0084] The test animals were anesthetized by intramuscular injection of xylazine (5 mg / kg) and ketamine (40 mg / kg), and anesthesia was maintained by intravenous drip of xylazine and ketamine (80 mg + 800 mg, prepared in 12 ml) via the marginal vein of the right ear of the rabbit at a rate of 5 ml / h. After exposing one common carotid artery surgically, 30 min after intravenous injection of the drug, a piece of filter paper (10 mm × 10 mm) on the strip was wound around the carotid artery. After winding, blood flow was not affected. The filter paper contained 100 μl of an aqueous solution of FeCl 2 with a concentration of 13%. After 5 min, the filter paper was removed and the blood vessel was rinsed twice with 0.9% sodium chloride injection. 30 min after using the filter paper, the injured carotid artery was excised, and the thrombus in the blood vessel was removed and weighed.

[0085] 3. Test results

[0086] As shown in Table 2, in the rabbit carotid artery thrombosis model induced by FeCl2, the weight of the arterial thrombus in the carotid artery of rabbits in the compound 1 group of the examples was significantly reduced compared with that in the carotid artery of rabbits in the control group 1, with statistical significance.

[0087] Table 2 Weight of arterial thrombus in the rabbit carotid artery thrombosis model induced by FeCl 2 (unit: mg)

[0088] Animal Number / Group Model Group Comparative Example 1 Group Compound 1 Group Mean 18.53 <![CDATA[4.80 +++ > <![CDATA[2.47 +++ Δ]]>

[0089] Compared with the model group: +++ P < 0.001: Compared with the control group 1: ΔP < 0.01.

[0090] The above embodiments are only one of the preferred embodiments of the present invention and should not be used to limit the protection scope of the present invention. Any modification or polishing made without substantial significance in the main design concept and spirit of the present invention, as long as the technical problems solved are still the same as those of the present invention, should be included in the protection scope of the present invention.

Claims

1. A compound of formula (I) or a pharmaceutically acceptable salt thereof: 。 2. The compound or pharmaceutically acceptable salt according to claim 1, wherein, the salt is a metal salt.

3. The compound or pharmaceutically acceptable salt according to claim 2, wherein, the metal salt is selected from sodium salt, potassium salt, calcium salt, lithium salt or magnesium salt.

4. A pharmaceutical composition comprising the compound or pharmaceutically acceptable salt according to any one of claims 1 to 3, wherein, the composition further comprises a pharmaceutically acceptable carrier and / or excipient.

5. A method for preparing the compound or pharmaceutically acceptable salt according to any one of claims 1 to 3, wherein, the method comprises the following route: 。 6. Use of the compound or pharmaceutically acceptable salt according to any one of claims 1 to 3 or the composition according to claim 4 in the preparation of a medicament for treating and / or preventing diseases related to the FⅪa receptor.

7. The use according to claim 6, wherein, the diseases related to the FⅪa receptor are selected from thrombosis or thromboembolism-related disorders.

8. The use according to claim 6, wherein, the diseases related to the FⅪa receptor are selected from cerebrovascular arterial diseases and / or peripheral arterial diseases.

9. The use according to claim 6, wherein, the diseases related to the FⅪa receptor are selected from transient ischemic attack, ischemic stroke and / or peripheral arterial diseases.

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