A novel carbamate compound and use thereof

Abstract

The present application relates to a kind of anti-platelet aggregation drugs and its use.The drug is the compound of structure I, or its pharmaceutically acceptable salt, solvate or deuteride;The compound of the present application has significant anti-platelet aggregation effect, more suitable water solubility and good stability, and is expected to become a new type of injectable anti-platelet aggregation drug.

Description

TECHNICAL FIELD

[0001] The present application belongs to the field of pharmaceutical chemistry, and particularly relates to a carbamate compound and use of the compound in preparation of an anti-platelet aggregation drug. BACKGROUND

[0002] It is well known in the art that platelet aggregation can cause a series of cardiovascular and cerebrovascular and other arterial circulatory disorders, including (1) acute coronary syndrome (ACS), such as unstable angina (UA), acute ST segment elevation myocardial infarction (STEMI) and acute non-ST segment elevation myocardial infarction (NSTEMI); (2) atherosclerotic diseases, such as myocardial infarction, ischemic stroke, peripheral arterial disease; (3) and thrombotic complications, etc. Among the current anti-platelet aggregation drugs, clopidogrel is of less risk of bleeding and is concerned, but due to the existence of "clopidogrel resistance", the drug's onset time and application scenarios, such as acute thrombosis, are greatly limited. Although the prior art also attempts to prepare an injection form of clopidogrel in order to achieve rapid onset and overcome the defects such as slow onset in acute treatment environment, such as the ASD-002 nanoemulsion of Ascendia Company, the MDCO-157 (cyclodextrin inclusion) injection of CyDex Company and the JIN-2013 nano-liposome injection of Jina Pharmaceuticals Company; but so far, due to the solubility and water, light and heat stability of clopidogrel, all have failed. In addition, the 2-oxo-clopidogrel developed by Chengdu Shibeikang Biomedical Technology Co., Ltd. overcomes the adverse reactions of "clopidogrel resistance" and has the advantages of rapid onset and higher bioavailability; but it is found in the research that 2-oxo-clopidogrel still has the defect of instability in water, which greatly challenges and limits the development into an injection form. Therefore, the development of derivatives of the intermediate metabolite 2-oxo-clopidogrel has attracted widespread attention from all sectors of society.

[0003] At present, only vecalogrel among the prodrug molecules based on 2-oxo-clopidogrel has entered the clinic, and is in the form of tablets. However, it is found in comparative studies that vecalogrel still has many limitations, such as unsatisfactory solubility and thermal stability, which will greatly limit its administration form and clinical application scenarios.

[0004] In summary, it is a difficult problem to be solved in the clinic to develop a prodrug molecule that can release clopidogrel active metabolite component (H4) in vivo for a long time and has good water solubility and stability, and at the same time solve the characteristics of clopidogrel that cannot be injected. SUMMARY

[0005] One of the purposes of the present application is to provide a 2-oxo-clopidogrel prodrug molecule compound with excellent solubility in water.

[0006] The second object of the present application is to provide a use for preventing and treating heart, brain and other arterial circulatory disorders caused by platelet aggregation.

[0007] The technical solution of the present application is as follows:

[0008] The present application provides a compound shown in formula I, or a pharmaceutically acceptable salt, solvate or deuteride thereof:

[0009] .

[0010] Further, the pharmaceutically acceptable salt includes a fumarate, an acetate, an ascorbate, a benzoate, a benzenesulfonate, a citrate, a hydrochloride, a hydrobromide, a maleate, a methanesulfonate, a sulfate, a bisulfate, a nitrate, an oxalate, a phosphate or a succinate.

[0011] Further, the pharmaceutically acceptable salt is selected from a fumarate, wherein the molar ratio of fumaric acid to the compound shown in formula I is 1:1 or 2:1.

[0012] The present application provides a pharmaceutical composition containing any of the above compounds, or a pharmaceutically acceptable salt, solvate or deuteride thereof, and the pharmaceutical composition further comprises a pharmaceutically acceptable excipient.

[0013] The present application also provides a use of the above compound, or a pharmaceutically acceptable salt, solvate or deuteride thereof, in the preparation of a medicament for preventing and / or treating heart, brain and other arterial circulatory disorders caused by platelet aggregation.

[0014] Further, the heart, brain and other arterial circulatory disorders caused by platelet aggregation include, but are not limited to, acute coronary artery syndrome, atherosclerotic disease, or thrombotic complications.

[0015] Further, the acute coronary artery syndrome includes, but is not limited to, angina pectoris or myocardial infarction.

[0016] Further, the acute coronary artery syndrome includes, but is not limited to, unstable angina (UA), acute ST segment elevation myocardial infarction (STEMI) or acute non-ST segment elevation myocardial infarction (NSTEMI).

[0017] Further, the atherosclerotic disease includes, but is not limited to, myocardial infarction, ischemic stroke or peripheral arterial disease.

[0018] Further, the ischemic stroke includes, but is not limited to, cerebral stroke.

[0019] Furthermore, the above-mentioned thrombotic complications include but are not limited to pulmonary infarction.

[0020] Beneficial technical effects:

[0021] The compounds of the present invention have better in vitro efficacy and superior pharmacokinetic characteristics. Specifically, the compounds of the present invention have a strong antiplatelet aggregation effect, rapid onset of action, high blood concentration and bioavailability of the pharmacokinetic in vivo, a longer half-life, and better efficacy. In addition, the compounds of the present invention have good in vitro solubility and high stability, low risk of hemolysis, no cardiotoxicity, and low risk of vascular irritation, and are highly safe, making them more suitable for formulation into injections, thus addressing clinical deficiencies. DETAILED DESCRIPTION

[0022] The present invention will be further described in detail below in conjunction with embodiments and test examples. The embodiments and test examples of the present invention are only used to illustrate the technical solutions 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 scope of protection of the present invention.

[0023] The compounds of the present invention, their stereoisomers, or pharmaceutically acceptable salts thereof can be prepared by selecting the synthetic routes described in the examples. Conventional conditions for the reaction starting materials and reaction solvents can be adjusted based on the desired substituents or salt formation. These conditions can be implemented by those skilled in the art based on the disclosure of the present invention. Furthermore, column chromatography in the present invention, unless otherwise specified, refers to silica gel column chromatography. The elution solvent, unless otherwise specified, can be determined using a combination of the reaction solvent, common knowledge, or commonly used methods employed by those skilled in the art, to determine a single or mixed elution solvent.

[0024] The structure of the compound was determined by NMR ( 1 H NMR) or liquid chromatography-mass spectrometry (LC-MS).

[0025] The liquid chromatography-mass spectrometer (LC-MS) was Agilent G6120B (used with Agilent 1260 liquid chromatography); the nuclear magnetic resonance instrument ( 1 HNMR) was Bruker AVANCE-400 or Bruker AVANCE-800, and nuclear magnetic resonance ( 1 H NMR) shift ( δ ) are given in parts per million (ppm), the solvent used for determination is DMSO-d6 or CDCl3, the internal standard is tetramethylsilane (TMS), and the chemical shift is in 10 -6 The units are given in ppm.

[0026] The term "room temperature" in the present invention refers to a temperature between 10 and 25°C.

[0027] Example 1

[0028] Preparation of (S)-5-(l-(2-chlorophenyl)-2-(methoxy-d3)-2-oxoethyl)-4,5,6,7- tetrahydrothieno[3,2-c]pyridin-2-yl-4-methylpiperazine-l-carboxylate (Compound 1)

[0029]

[0030] Step 1: Preparation of (R)-methyl 2-(2-chlorophenyl)-2-hydroxyacetate (Intermediate b)

[0031] Into a 50 mL three-necked flask, 4 g of (R)-o-chloromandelic acid was added, and 12 mL of anhydrous deuterated methanol was added to dissolve the mixture by stirring. Then, 0.17 g of concentrated sulfuric acid was slowly added dropwise. After the dropwise addition was completed, the temperature was raised to 78°C by heating with an oil bath, and the reaction was carried out at reflux for 2 hours. The reaction was monitored by TLC (developing solvent: ethyl acetate / petroleum ether = 1 / 3, coloration under a 254 nm UV lamp). The methanol was removed by concentration under reduced pressure at an external temperature of 45°C, and the residue was dissolved by stirring in 12 mL of ethyl acetate. The ethyl acetate layer was washed twice with 10 mL of water, and then washed with 10 mL of saturated sodium bicarbonate solution and 10 mL of saturated brine, respectively. The ethyl acetate layer was dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure at an external temperature of 45°C to remove the ethyl acetate, and 0.415 g of a slightly yellow transparent oily liquid was obtained, which was Intermediate b. Yield: 95.1%.

[0032] ESI-MS: m / z = 204.1 (M+H) + .

[0033] 1 HNMR (400 MHz, CDCl3) δ:7.4 (m,1H),7.35 (m, 1H), 7.23 (m, 2H), 5.58(s, 1H), 4.06 (brs, 1H).

[0034] Step 2: Preparation of (R)-methyl 2-(2-chlorophenyl)-2-((4-nitrophenyl)sulfonyl)oxyacetate (Intermediate d)

[0035] Into a 30 mL single neck flask, 4.15 g of intermediate I and 15 mL of dichloromethane were added at room temperature, and the mixture was stirred to dissolve under ice-bath. Then, 2.51 g of triethylamine was added, and the temperature was controlled to be lower than 5°C. 4.35 g of p-nitrobenzenesulfonyl chloride dissolved in 15 mL of dichloromethane was added dropwise at a constant pressure, and the dropping was completed after 1.5 hours. After the dropping, the ice-bath was maintained for 0.5 hours. Then, 5 mL of 4M hydrochloric acid was added dropwise at a constant pressure under ice-bath, and the stirring was continued until the solution became yellow. Then, the mixture was separated. The dichloromethane layer was washed with 10 mL of water and 10 mL of saturated brine, and then, the dichloromethane was removed under reduced pressure at 35°C to obtain a yellow paste of intermediate d. The crude product was added into a 50 mL beaker, and then, 10 mL of ice-methanol was added under ice-bath for 0.5 hours. During the process, a large amount of white solid was precipitated. The solid was filtered to obtain 5.61 g of white powder, which was intermediate d, and the yield was 70.8%.

[0036] ESI-MS: m / z = 389.1 (M+H) + .

[0037] 1 HNMR (400 MHz, CDCl3) δ: 8.31 (d, J = 8.9 Hz,2H),8.07 (d,J = 8.9 Hz,2H), 7.31 (m, 4H), 6.39 (s, 1H).

[0038] Step 3: Preparation of (S)-methyl 2-(2-chlorophenyl)-2-((S)-2-oxo-2,6,7,7a-tetrahydrothieno[3,2-c]pyridin-5(4H)-yl)acetate-d3 (intermediate f)

[0039] Reaction: 5.61 g of intermediate b, 3.19 g of compound e, 5.39 g of anhydrous sodium carbonate and 100 mL of acetonitrile were added into a 200 mL three-neck flask at room temperature, and then, the mixture was stirred at 38°C for 5 hours under nitrogen. The reaction solution was filtered through diatomite, and then, the acetonitrile was removed under reduced pressure at 45°C. The residue was dissolved in 20 mL of dichloromethane, and then, the dichloromethane layer was washed with 10 mL of water and 10 mL of saturated brine, respectively. The mixture was separated, and then, the filtrate was concentrated under reduced pressure to obtain 4.18 g of partially crystallized brown paste semi-solid. Then, 4.18 g of sbk002 crude product and 10 mL of acetone were added into a 20 mL beaker under 60°C water-bath, and then, the mixture was stirred for 10 minutes. The solution was filtered while hot, and then, the crystallization was performed to obtain 2.95 g of white crystalline powder, and the yield was 60.0%.

[0040] ESI-MS: m / z = 341.1 (M+H) + .

[0041] Step 4: Preparation of (S)-5-(l-(2-chlorophenyl)-2-(methoxy-d3)-2-oxoethyl)-4,5,6,7- tetrahydrothieno[3,2-c]pyridin-2-yl-4-methylpiperazine-1-carboxylate (Compound 1)

[0042] 50 ml three-necked flask was added (2S)-2-(2-oxo-7,7a-dihydrothieno[3,2-c]pyridin-5(2H,4H,6H)-yl)-2-(2-chlorophenyl)-acetic acid methyl ester (1 g, 3 mmol) and 4-methylpiperazine-1-carboxylic acid chloride hydrochloride (884 mg, 4.44 mmol), DMF (20 ml), stirred and cooled to 0°C, dropwise added DBU (1.37 g, 9 mmol). After dropwise addition, the reaction was completed at room temperature for 2 h. Added EA, washed with water twice, and brine once. The organic phase was dried with anhydrous Na2SO4and concentrated. The concentrate was separated and purified by column chromatography, the product was collected and concentrated to obtain 1.2 g of Compound 1, with a yield of 87.6% and a purity of 98.34%.

[0043] ESI-MS: m / z = 467.1 (M+H) + .

[0044] 1 HNMR (400 MHz, DMSO-d6) δ: 7.60~7.54 (m, 1H), 7.51~7.46 (m, 1H), 7.46~7.35 (m, 2H), 6.39 (s, 1H), 4.86 (s, 1H), 3.57 (s, 2H), 3.40 (s, 2H), 2.80 (qt, 2H), 2.63 (d, 2H), 2.48~2.39 (m, 6H), 2.34 (s, 3H).

[0045] Example 2

[0046] Preparation of (S)-5-(l-(2-chlorophenyl)-2-(methoxy-d3)-2-oxoethyl)-4,5,6,7- tetrahydrothieno[3,2-c]pyridin-2-yl-4-methylpiperazine-1-carboxylate fumarate (Compound 2)

[0047]

[0048] 25 ml flask was added (S) -5- (1- (2-chlorophenyl) -2- (methoxy-d3) -2-oxoethyl) -4, 5, 6, 7-tetrahydrothieno [3, 2-c] pyridin-2-yl-4-methylpiperazine-1-carboxylate (800 mg, 1.71 mmol), EA (2.5 ml) was dissolved, fumaric acid (198 mg, 1.71 mmol) was added and stirred, a large amount of solid was precipitated. Stirring for 3 h, filtering, drying to obtain 240 mg of the title compound. Yield 56.2%, purity 98.81%.

[0049] ESI-MS: m / z = 467.1 (M+H) + .

[0050] 1 HNMR (400 MHz, DMSO-d6) δ: 13.82~9.71 (br, 2H), 7.64~7.56 (m, 1H),7.56 ~7.47 (m, 1H), 7.46~7.35 (m, 2H), 6.62 (d, 2H),6.39 (s, 1H), 4.86 (s,1H), 3.67 (s, 3H), 3.57 (s, 2H), 3.46 (s, 2H), 2.82 (qt, 2H), 2.69 (d, 2H),2.48~2.39 (m, 6H), 2.30 (s, 3H)。

[0051] Comparative Example 1: 2-oxo-clopidogrel (metabolic intermediate in clopidogrel activation process)

[0052]

[0053] Prepared by Chengdu Shibeikang Biomedical Technology Co., Ltd., ee = 98.8%.

[0054] Comparative Example 2: Vicagrel (in clinical studies)

[0055]

[0056] Prepared according to CN201010624329.7, purity 98.22%.

[0057] Test Example 1: Anti-platelet aggregation effect study

[0058] Test purpose: to evaluate and compare the therapeutic effects of each compound on anti-platelet aggregation after the same molar administration.

[0059] Test method

[0060] (1) Grouping

[0061] Forty rats were randomly and evenly divided into four groups: solvent control group, Comparative Example 1 (2-oxo clopidogrel) group, Comparative Example 2 (vaccagrel) group, and Compound 1 group.

[0062] (2) Anesthesia induction

[0063] The rats were anesthetized by intraperitoneal injection of 1.5% sodium pentobarbital.

[0064] (3) Fixation

[0065] The anesthetized animals were transferred to the operating table, and the eyelid reflex and pain sensation of the rats were observed. After the eyelid reflex and pain sensation of the limbs and tail disappeared, the operation could be started.

[0066] (4) Rat abdominal aorta blood sampling

[0067] After the anesthetic was injected, until the whole body was soft, the rat could be fixed on the operating table in a supine position. After routine disinfection, the abdominal cavity was cut along the midline with surgical scissors, the fat around the blood vessels was gently scraped with small forceps, and the excess fat covering the blood vessels was wiped clean with a cotton ball until the blood vessels were clearly visible (the abdominal aorta is located above the spine; the abdominal vein is thicker and darker than the abdominal aorta). The blood vessels were fixed to avoid displacement. The thumb and index finger of the left hand fixed the fat and other organs on both sides of the blood vessels, the ring finger pressed the upper end of the blood vessel puncture point, the blood pressure was lowered to avoid blood spitting, the right hand held the puncture needle with the bevel facing down at an angle of about 30 degrees, and the needle was inserted towards the heart end at a depth of about 5 mm. After the needle end bled, the other end of the blood sampling needle was inserted into the vacuum tube. After the needle was inserted, the needle was clamped with a hemostat to prevent the blood vessels from being punctured by the needle due to struggling caused by insufficient anesthesia.

[0068] (5) Platelet preparation

[0069] Three hours after administration, the rats were anesthetized with sodium pentobarbital, and blood was collected from the abdominal aorta. The blood was anticoagulated with 3.8% sodium citrate 1:9, mixed well, and centrifuged at 200 g for 10 min. The supernatant was platelet-rich plasma (PRP). The remaining plasma was centrifuged at 1600 g for 15 min, and the supernatant was platelet-poor plasma (PPP).

[0070] (6) ADP-induced platelet aggregation

[0071] The platelet aggregation rate was measured using a platelet aggregometer (Helena, USA, model: Agg RAM). First, the PPP corresponding to the PRP to be tested in each channel was corrected for light transmittance. After correction, the PPP was removed, and the cuvette containing 225 μl of PRP to be tested was placed in each channel, a stirrer was added, ADP (final concentration 20 μM) was added, and the platelet aggregation rate detection was started immediately.

[0072] Test results

[0073] The platelet aggregation rate data of each group after the same molar administration are shown in Table 1.

[0074] The results show that: (1) the platelet aggregation rate (%) of the solvent control group is 76.06±3.29; the platelet aggregation rates (%) of the Comparative Example 1 group, the Comparative Example 2 group and the Compound 1 group are 31.32±6.07, 39.32±12.38 and 19.06±5.47, respectively, and each of the administration groups has a significant inhibitory effect on ADP-induced platelet aggregation of rats compared with the solvent control group (all P<0.01).

[0075] (2) Compared with the Comparative Example 2 group, the Compound 1 group has a significant advantage in inhibiting ADP-induced platelet aggregation of rats (P<0.01), which has statistical significance.

[0076] (3) From the test results, it can be seen that under the same dose, after single intravenous injection administration, the Compound 1 in the present application has a significantly better inhibitory effect on ADP-induced platelet aggregation of rats than the Comparative Example 2 compound and the Comparative Example 1 compound.

[0077]

[0078] Test Example 2: Effect of the compound on the stability of overexpressed hERG channel current

[0079] Test sample

[0080] Compound 1, control compound Cisapride.

[0081] Test method

[0082] The manual patch clamp technique (the gold standard for hERG safety evaluation) was used to study the inhibitory effect of the test compound 1 on the hERG potassium channel and to evaluate the risk of inducing ventricular repolarization toxicity.

[0083] Test results

[0084] In this study, the concentration-effect relationship of the test compound 1 on the hERG channel was detected by the manual patch clamp technique, so as to evaluate the risk of the test substance on the inhibition of the hERG potassium channel of the heart. The test results are shown in Table 2.

[0085]

[0086] The above test results show that the compound of the present application has a small risk of inducing ventricular repolarization toxicity and high safety.

[0087] Test Example 3: Pharmacokinetic study

[0088] Test purpose

[0089] The pharmacokinetic characteristics of each compound after a single intragastric administration in rats at the same molar dose were investigated, and the main pharmacokinetic parameters were compared.

[0090] Materials and methods

[0091] (1) Test substances

[0092] Comparative Example 1 (2-oxo-clopidogrel), Compound 1.

[0093] (2) Preparation of administration preparations

[0094] The test compounds were accurately weighed into clean administration containers, an appropriate amount of Solutol was added to dissolve them, and the containers were vortexed and shaken, pure water was added, and the containers were ultrasonicated, vortexed and shaken until the compounds were completely dissolved; the administration preparations were freshly prepared on the day of administration.

[0095] (3) Test grouping and administration

[0096] Twelve healthy adult SD rats were administered by gavage. The specific scheme is shown in Table 3.

[0097]

[0098] (4) Test method

[0099] Grouping and fasting: SD rats were randomly grouped, with 6 rats in each group, and the corresponding compounds were administered by gavage according to Table 3.

[0100] Sample collection and processing: 0.2 ml of blood was collected at different time points before administration (0 h) and after administration 5 min, 15 min, 30 min, 1 h, 2 h, 4 h, 8 h, and 24 h, and after anticoagulation with EDTA-K2, the plasma was centrifuged at 4°C for 5 min and stored at -80°C for testing.

[0101] Detection: LC / MS / MS was used to detect the blood drug concentration of 2-oxo-clopidogrel in all PK plasma samples, and WinNonlin 7.2 software was used to calculate the pharmacokinetic parameters of each compound.

[0102] Test results

[0103]

[0104] After equimolar administration, the T 1 / 2 , C max , and AUC 0-last of Compound 1 were significantly prolonged and increased compared with those of Comparative Example 1, with statistical significance (P < 0.01). This indicates that the absorption of Compound 1 in rats is significantly better than that of Comparative Example 1, and it has a more optimal pharmacokinetic characteristic.

[0105] In summary, the compound of the present application is better absorbed, has high bioavailability, and is expected to fully exert the drug efficacy.

[0106] Test Example 4: solubility study

[0107] Accurately weigh the sample of Example Compound 2 and Comparative Example 2 (Vorapaxar), and take 1 ml of each in physiological saline and pH 1.2 buffer at 25-30°C, and then add 10 mg of each compound, respectively, to measure the solubility data as shown in the following table: The compound 2 of the present application has excellent solubility, which can meet the requirement of preparing an injection solution with effective concentration in clinic, while the solubility of Comparative Example 2 is extremely low, which does not meet the requirement of injection solution. If other solubilizers are added, potential safety risks may be caused.

[0108]

[0109] Test Example 5: study on influencing factors

[0110] Test method: weigh the sample of Compound 2 and Comparative Example 2 (Vorapaxar) respectively and place them in a weighing bottle, and then place them under high temperature (60°C), high humidity (RH 80%) and light (5000 Lux) conditions, respectively.

[0111] The test results of Compound 2 are shown in the following table: under high temperature, high humidity and light conditions for 10 days, the content change of Compound 2 is small, and the appearance shape has no obvious change, and the stability is still good.

[0112]

[0113] However, Comparative Example 2 was placed under high temperature (60°C) for two days, and the property changed greatly, from white solid to yellow-brown oily substance, indicating that Comparative Example 2 has poor stability under high temperature conditions.

[0114] In summary, the compound of the present application has temperature, humidity and light stability, and is better than Comparative Example 2.

[0115] Those skilled in the art can make various modifications and changes to the compound, composition and method of the present application without departing from the spirit of the present application, and these all belong to the same or equivalent range of the present application.

Claims

1. A compound represented by Formula I: ###0001### or a pharmaceutically acceptable salt thereof. 。 2. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein, the pharmaceutically acceptable salt is selected from the group consisting of a fumarate, an acetate, an ascorbate, a benzoate, a besylate, a citrate, a hydrochloride, a hydrobromide, a maleate, a mesylate, a sulfate, a bisulfate, a nitrate, an oxalate, a phosphate, or a succinate.

3. The compound of claim 2, or a pharmaceutically acceptable salt thereof, wherein, the pharmaceutically acceptable salt of the compound is a fumarate, wherein the molar ratio of fumaric acid to the compound represented by I is 1:1 or 2:

1.

4. A pharmaceutical composition comprising a compound of any one of claims 1-3, or a pharmaceutically acceptable salt thereof. the pharmaceutical composition further comprises a pharmaceutically acceptable excipient.

5. Use of the compound of any one of claims 1 to 3, or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of claim 4, in the manufacture of a medicament for preventing and / or treating a heart, brain, and other arterial circulatory disorder caused by platelet aggregation.

6. Use according to claim 5, characterized in that, the heart, brain, and other arterial circulatory disorder caused by platelet aggregation is selected from the group consisting of acute coronary syndrome, atherosclerotic disease, or a thrombotic complication.

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