Oxo-pyridazine compounds, their preparation methods and uses

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 remarkable curative effects.

Description

Technical Field

[0001] The present invention relates to the field of medicinal chemistry, and particularly relates 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 tens of millions of lives 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, thereby 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 difficult 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 safety, effectiveness, 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. Most of the compounds in the examples exhibit 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 pharmaceutically acceptable salts:

[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 the other hand, 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 stereoisomer or pharmaceutically acceptable salt or its composition 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 cerebrovascular arterial diseases include but are not limited to transient ischemic attack (TIA), ischemic stroke, or events originating from thrombosis and / or thromboembolism that cause stroke or TIA; the 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 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 cardiogenic stroke includes but is not limited to stroke caused by atrial fibrillation; the 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 FeCl2-induced rabbit carotid artery thrombosis model, the weight of the arterial thrombus in the carotid artery of rabbits in the compound 1 group of the examples is significantly reduced compared with the weight of the arterial thrombus in the carotid artery of rabbits in the comparative example 1 group, which is statistically significant and the drug effect is 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 1H 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 Agilent 1260); the nuclear magnetic resonance spectrometer ( 1 1HNMR) is Bruker AVANCE-400 or Bruker AVANCE-800. The nuclear magnetic resonance ( 1 1H NMR) 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)-3-methoxy-6-oxopyridazin-1(6H)-yl)butanamido)-2-fluoro-N-(methyl-d3)benzamide (Compound 1):

[0025]

[0026] Step 1: Preparation of intermediate b

[0027] Dissolve diisopropylamine (24 mmol) in 25 ml of THF, cool to below -60 °C, and add dropwise 9.14 ml of a 2.5 M n-butyllithium n-hexane solution over about 1 hour. After addition, stir the reaction at -60 °C for 15 minutes. Then add dropwise a 10 ml THF solution of compound a (20 mmol). After addition, stir the reaction at -60 °C for 2 hours. Add dropwise triisopropyl borate (22 mmol). After addition, slowly warm up to room temperature (20 °C) and stir the reaction for 30 minutes. Add dropwise a mixture of 4 g of acetic acid and 21 g of water to terminate the reaction. After addition, stir at room temperature for 30 minutes. Evaporate 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 a solid. The yield is 70.20%, and the HPLC purity is 98.43%.

[0028] ESI-MS: m / z = 185.1 (M+H)+.

[0029] Step 2: Preparation of intermediate e

[0030] Take compound d (10.0 mmol), Pd(amphos)Cl2 (0.15 mmol) and suspend them in 25 ml of tert-amyl alcohol, heat to 85 °C, and add dropwise a mixed solution of compound c (1.2 mmol), sodium carbonate (30 mmol) and 25 ml of water. After addition, 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 intermediate e. The yield is 77.99%, and the HPLC purity is 98.27%.

[0031] ESI-MS: m / z = 319.1 (M+H)+.

[0032] Step 3: Preparation of intermediate f

[0033] Take compound e (6 mmol), anhydrous lithium chloride (30 mmol), p-toluenesulfonic acid monohydrate (12 mmol) and mix them with 20 ml of isopropyl alcohol, heat under reflux for 16 hours. Cool to room temperature, evaporate half of the solvent, add 30 ml of 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 intermediate f in solid form. The yield is 83.51%, and the HPLC purity is 97.14%.

[0034] ESI-MS: m / z = 305.1 (M+H)+.

[0035] Step 4: Preparation of intermediate h

[0036] Add compound f (2 mmol), tetramethylguanidine (7 mmol), 6 ml of isopropanol, and 1.5 ml of acetone to a 25-ml reaction flask. Stir at room temperature for 15 minutes, then add compound g (2.2 mmol) and stir the reaction overnight at room temperature. The next day, add water to terminate the reaction, extract with EA, separate the aqueous layer, and wash the organic layer successively with saturated ammonium chloride, water, saturated brine, dry over anhydrous sodium sulfate, filter, evaporate the solvent, purify by column chromatography, collect the product to obtain the pure product. The yield is 84.71% and the purity is 97.49%.

[0037] ESI-MS: m / z = 584.2 (M+H)+.

[0038] Step 5: Preparation of intermediate j

[0039] Add compound h (2 mmol) and 16 ml of methanol to a 25-ml reaction flask, stir to dissolve, and cool to 0 °C. Weigh lithium hydroxide monohydrate (4 mmol) and dissolve it in 8 ml of water, then add it dropwise to the reaction flask. After adding, react at room temperature for 2 h. Add water to terminate the reaction, adjust the pH to weakly acidic with 5% citric acid, extract with EA, separate the aqueous layer, wash the organic layer successively with water, saturated NaCl, dry over anhydrous sodium sulfate, filter, evaporate the solvent, perform column chromatography, and collect the target product to obtain intermediate j. The yield is 81.86% and the purity is 98.58%.

[0040] ESI-MS: m / z = 528.1 (M+H)+.

[0041] Step 6: Preparation of (S)-4-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-3-methoxy-6-oxo-pyridazin-1(6H)-yl)butanamido)-2-fluoro-N-(methyl-d3)benzamide (Compound 1)

[0042] Add intermediate j (1 mmol), 3 ml of DMF, deuterated methylamine hydrochloride (1.1 mmol), EDCI (2 mmol), and HOBT (2 mmol) to a 50-ml reaction flask. Cool to 0 °C and add DIPEA (4 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 Compound 1. The yield is 77.54% and the purity is 98.40%.

[0043] ESI-MS: m / z = 544.2 (M+H)+.

[0044] 11H NMR (400 MHz, DMSO-d6) δ: 10.76 (s, 1H), 9.12 (s, 1H), 8.10 (s, 1H), 7.90–7.76 (m, 2H), 7.71–7.60 (m, 2H), 7.37 (dd, J = 8.6, 2.0 Hz, 1H), 7.13 (s, 1H), 6.54 (s, 1H), 5.52 (dd, 1H), 3.26 (s, 3H), 2.10 (m, 2H), 0.79 (t, J = 7.2 Hz, 3H).

[0045] 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

[0046]

[0047] Synthesized according to the method described in Patent CN108026072B, purity: 98.5%.

[0048] ESI-MS: m / z = 593.1 (M+H) + .

[0049] 1 1H 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).

[0050] Test Example 1: Inhibitory effect on coagulation factor FXIa

[0051] 1. Test samples

[0052] Compound 1 of the example and Comparative Example 1.

[0053] 2. Test procedure

[0054] 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.

[0055] 2) Prepare a 10X working solution of the compound.

[0056] 3) Prepare 0.8 nM Human FXIa working solution (2X), mix well and set aside for use.

[0057] 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.

[0058] 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.

[0059] 6) Prepare 750 μM S-2366 working solution (2.5X), mix well and set aside for use.

[0060] 7) Add 16 μL of the S-2366 working solution in step 6) to all 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.

[0061] 8) After incubation, use EnVision to read the absorbance at OD405 nm and collect the data.

[0062] Set 5 concentrations, namely: 200 nM, 40 nM, 8 nM, 1.6 nM, 0.32 nM, to detect the IC 50 value.

[0063] 3. Data analysis

[0064] 1) Z’factor = 1 - 3*(SD Max + SD Min ) / (Mean Max - Mean Min );

[0065] 2) CV Max = (SD Max / Mean Max )*100%;

[0066] 3) CVMin = (SD Min / Mean Min )*100%;

[0067] 4) S / B = Singal / Background;

[0068] 5) Blank control: 0.1% DMSO; Positive control: Comparative Example 1;

[0069] 6) IC 50Calculation formula: Y = Bottom + (Top - Bottom) / (1 + 10^((LogIC 50 -X) * HillSlope)).

[0070] X: Log value of compound concentration; Y: Inhibition%.

[0071] 4. Test results

[0072] The test results are shown in the table: Under the condition of the same molar concentration, the in vitro inhibitory activity of Compound 1 of the present invention against FXIa is comparable to that of the compound in Comparative Example 1.

[0073] Table 1 Determination of the activity of coagulation factor FXIa inhibitor (IC50)

[0074] Sample <![CDATA[IC 50 (nM)]]> Sample <![CDATA[IC 50 (nM)]]> Compound 1 0.8 Comparative Example 1 7.6

[0075] Test Example 2: In vivo pharmacodynamic evaluation of a rabbit arteriovenous shunt model

[0076] 1. Test samples

[0077] Compound 1 of the example and Comparative Example 1.

[0078] 2. Test methods

[0079] 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.

[0080] The Comparative Example 1 group and the Compound 1 group were respectively administered the compounds shown in Comparative Example 1 and Compound 1 by single intravenous injection at a dose of 6 mg / kg.

[0081] Anesthetize the test animals by intramuscular injection of xylazine (5 mg / kg) and ketamine (40 mg / kg), and maintain the anesthetic effect by intravenous drip of xylazine and ketamine (80 mg + 800 mg, prepared into 12 ml) through the right ear marginal vein of the rabbit (5 ml / h). Expose one common carotid artery surgically. 30 minutes after intravenous injection of the drug, use a piece of filter paper (10 mm × 10 mm) on the strip to wrap the carotid artery. After wrapping, the blood flow is not affected. The filter paper contains 100 μl of an aqueous solution of FeCl2 with a concentration of 13%. After 5 minutes, remove the filter paper and rinse the blood vessel 2 times with 0.9% sodium chloride injection. 30 minutes after using the filter paper, excise the injured carotid artery, remove the thrombus in the blood vessel and weigh it.

[0082] 3. Test results

[0083] 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 the rabbits in the group of Example Compound 1 was significantly reduced compared with that in the group of Comparative Example 1, and the difference was statistically significant.

[0084] Table 2 Weight of arterial thrombus in the rabbit carotid artery thrombosis model induced by FeCl2 (unit: mg)

[0085] Animal Number / Group Model Group Comparative Example 1 Group Compound 1 Group Mean 18.53 <![CDATA[4.80 *** > <![CDATA[2.39 ***Δ >

[0086] Compared with the model group: *** P < 0.001; compared with the group of Comparative Example 1: Δ P < 0.05.

[0087] 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 modifications or polishings made without substantial significance in the main design concept and spirit of the present invention, as long as the technical problems solved are still consistent with 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, characterized in that, The metal salt is selected from sodium salt, potassium salt, calcium salt, lithium salt or magnesium salt.

4. A pharmaceutical composition containing the compound or pharmaceutically acceptable salt according to any one of claims 1 to 3, characterized in that, The composition further contains 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, characterized in that, The method includes 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 application according to claim 6, characterized in that, The diseases related to the FⅪa receptor are selected from thrombosis or thromboembolism-related disorders.

8. The application 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 application according to claim 6, wherein The diseases related to the FⅪa receptor are selected from transient ischemic attack, ischemic stroke, peripheral arterial occlusion or stent thrombosis.

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