Carbamate fumarate, crystalline form and amorphous form thereof, and preparation method therefor

Abstract

Provided are novel carbamate fumarate, a crystalline form and amorphous form thereof, and a preparation method therefor. The carbamate fumarate and the crystalline form and amorphous form thereof have high stability and high solubility, low hygroscopicity, and improved pharmaceutical properties, and achieves a better anti-platelet aggregation effect and higher efficacy, thereby facilitating the clinical application of drugs.

Description

A carbamate fumarate, its crystalline form, amorphous form, and preparation method thereof.

[0001] This application claims priority to the invention patent filed on November 22, 2024, entitled "A carbamate fumarate and its crystalline, amorphous and preparation methods", patent number CN202411684869.2, which is hereby expressly incorporated herein by reference. Technical Field

[0002] This invention relates to the field of medicinal chemistry, specifically to the salt form, crystalline form, and amorphous form of a novel carbamate compound, its uses, and preparation methods. Background Technology

[0003] Platelet aggregation can trigger a series of cardiovascular, cerebrovascular, and other arterial circulatory disorders, mainly including acute coronary syndrome (ACS), atherosclerotic diseases, and thrombotic complications. Currently, the design of prodrug molecules based on clopidogrel's metabolic intermediate 2-oxoclopidogrel is very popular. For example, patent CN116262757A describes a carbamate compound, in which compound 1 with the following structure and its salt have achieved more superior antiplatelet aggregation effects and in vivo pharmacokinetic properties.

[0004] However, further research revealed that this carbamate compound still has many limitations, such as incompatibility between thermal stability and solubility (it cannot be achieved simultaneously), and the efficacy needs to be further improved, which greatly limits the form of administration and clinical application scenarios.

[0005] Based on this, developing more advantageous salt forms and crystal forms to improve efficacy and broaden pharmaceutical applications, providing reliable options for clinical use and achieving a relatively superior clinical advantage, is a technical challenge that urgently needs to be solved for this class of drugs. Summary of the Invention

[0006] To address at least one of the technical problems existing in the prior art, this invention provides a novel carbamate fumarate, as well as its crystalline and amorphous forms. The carbamate fumarate of this invention, in both its crystalline and amorphous forms, exhibits high stability and solubility, good hygroscopicity, higher efficacy, ease of purification, suitability for large-scale production, and is beneficial for pharmaceutical formulation and clinical application.

[0007] On one hand, the present invention provides a carbamate fumarate as shown in formula (1):

[0008] The present invention also provides a crystalline form A of the carbamate fumarate as described above, which, when irradiated with Cu-Kα, exhibits X-ray powder diffraction patterns with diffraction peaks at the following 2θ values: 3.58±0.2°, 7.17±0.2°, 10.38±0.2°, 14.04±0.2°, 16.75±0.2°, 17.84±0.2°, and 18.49±0.2°.

[0009] Furthermore, the X-ray powder diffraction pattern of the crystal form A also includes any one or more diffraction peaks with 2θ values ​​as follows: 11.82±0.2°, 12.98±0.2°, 22.96±0.2°, 23.21±0.2°, 23.45±0.2°, 25.27±0.2°.

[0010] Furthermore, the X-ray powder diffraction pattern of crystal form A basically includes any or more of the following diffraction peaks with 2θ values: 3.58±0.2°, 4.48±0.2°, 7.17±0.2°, 7.91±0.2°, 10.38±0.2°, 11.82±0.2°, 12.98±0.2°, 14.04±0.2°, 14.31±0.2°, 15.11±0.2°. 2°, 15.51±0.2°, 16.24±0.2°, 16.75±0.2°, 16.98±0.2°, 17.84±0.2°, 18.49±0.2°, 19.35±0.2°, 19.81±0.2°, 20.29±0.2°, 21.60±0.2°, 22.96±0.2°, 23.21±0.2°, 23.45±0.2° °, 23.75±0.2°, 24.08±0.2°, 24.53±0.2°, 24.93±0.2°, 25.27±0.2°, 25.85±0.2°, 26.27±0.2°, 26.61±0.2°, 26.91±0.2°, 27.38±0.2°, 28.27±0.2°, 28.97±0.2°, 29.49±0.2° 30.49±0.2°, 31.33±0.2°, 32.49±0.2°, 32.93±0.2°, 33.81±0.2°, 34.82±0.2°, 35.79±0.2°, 36.17±0.2°, 36.54±0.2°, 37.44±0.2°, 38.10±0.2°, 38.69±0.2°, 39.61±0.2°.

[0011] Furthermore, the X-ray powder diffraction pattern of crystal form A described above contains the following 2θ values ​​and diffraction peaks with relative intensities:

[0012] Furthermore, the X-ray powder diffraction pattern of crystal form A also contains one or more of the following sets of 2θ values ​​and diffraction peaks with their relative intensities:

[0013] Furthermore, the X-ray powder diffraction pattern of crystal form A basically contains one or more sets of diffraction peaks with 2θ values ​​and their relative intensities as follows:

[0014] Furthermore, the crystal form A described above has an X-ray powder diffraction pattern as shown in Figure 1.

[0015] Furthermore, the differential scanning calorimetry (DSC) spectrum of the crystal form A exhibits an endothermic peak at 150℃±5℃; preferably, the crystal form A has a differential scanning calorimetry curve that is essentially as shown in Figure 2.

[0016] Furthermore, the crystal form A described above has a thermogravimetric analysis spectrum as shown in Figure 3.

[0017] The present invention also provides an amorphous form of the carbamate monofumarate as described above, which, when irradiated with Cu-Kα, exhibits a diffuse peak X-ray powder diffraction pattern; preferably, the amorphous form has an X-ray powder diffraction pattern substantially as shown in Figure 4.

[0018] In a second aspect, the present invention provides a method for preparing carbamate fumarate or its crystal form A as described above, comprising the following steps: (S)-5-(1-(2-chlorophenyl)-2-methoxy-2-oxoethyl-4,5,6,7-tetrahydrothieno[3,2-c]pyridin-2-yl-4-methylpiperazine-1-carboxylic acid ester reacts with fumaric acid under protic solvent conditions to form a salt; after the reaction is complete, the temperature is lowered, a solid precipitates, and the solid is filtered and dried to obtain carbamate fumarate crystals; wherein the protic solvent is selected from alcohol or acetonitrile; preferably, the alcohol is selected from methanol, ethanol or isopropanol.

[0019] The present invention also provides a method for preparing the amorphous form of the carbamate fumarate as described above, comprising the following steps: adding the above carbamate fumarate crystal A to a solvent, filtering after the solid dissolves, and spray-drying or freeze-drying the filtrate to obtain a white solid; wherein the solvent is selected from tert-butanol or water.

[0020] Thirdly, the present invention provides a pharmaceutical composition comprising any of the above-mentioned carbamate fumarate or its crystal form A or amorphous form, and further comprising a pharmaceutically acceptable carrier.

[0021] The present invention also provides a pharmaceutical formulation comprising any of the above-mentioned carbamate fumarate or its crystal form A or amorphous form, wherein the pharmaceutical formulation is selected from oral formulations or liquid formulations; preferably, the liquid formulation comprises an injection solution or a lyophilized formulation.

[0022] Fourthly, the present invention provides the use of any of the above-mentioned carbamate fumarate or its crystal form A, amorphous form, pharmaceutical composition or pharmaceutical preparation in the preparation of a medicament for the prevention and / or treatment of cardiovascular, cerebrovascular and other arterial circulatory disorders caused by platelet aggregation; preferably, the cardiovascular, cerebrovascular and other arterial circulatory disorders caused by platelet aggregation include, but are not limited to, acute coronary syndrome, atherosclerotic disease or thrombotic complications.

[0023] Furthermore, the atherosclerotic diseases include myocardial infarction, ischemic stroke, and peripheral artery disease. The acute coronary syndromes include non-ST-segment elevation acute coronary syndromes (including unstable angina or non-Q-wave myocardial infarction) and are used for ST-segment elevation acute coronary syndromes.

[0024] The term "pharmaceutically acceptable carrier" refers to a diluent, excipient, or carrier that is administered co-administered with the active ingredient and is suitable, to the extent of reasonable medical judgment, for contact with human and / or other animal tissues without excessive toxicity, irritation, allergic reactions, or other problems or complications commensurate with a reasonable benefit / risk ratio.

[0025] Note: Unless otherwise stated by evidence to the contrary or otherwise, the carbamate fumarate shown in formula (1) of the present invention represents a mass molar ratio of carbamate to fumaric acid of 1:1; the mass molar ratio in the corresponding crystalline and amorphous forms is also 1:1.

[0026] Compared with the prior art, the present invention has the following beneficial effects:

[0027] The carbamate fumarate of the present invention, whether in crystalline or amorphous form, is suitable for development into a drug with clinical application value, specifically in the following ways: (1) it has a good pharmaceutical solid form, high solubility, no or slight hygroscopicity, high stability, and is easy to transport and store; (2) the production process is simple, the materials are cheap and readily available, it is easy to scale up production, the production cycle is short, and it is energy-saving and environmentally friendly; (3) it has a better anti-platelet aggregation effect and is suitable for development into a drug for the prevention and / or treatment of cardiovascular, cerebrovascular and other arterial circulatory disorders caused by platelet aggregation. Attached Figure Description

[0028] Figure 1 shows the X-ray powder diffraction (XRPD) pattern of crystal form A of urethane fumarate;

[0029] Figure 2 shows the differential scanning calorimetry (DSC) curve of urethane fumarate crystal form A;

[0030] Figure 3 shows the thermogravimetric analysis (TGA) diagram of crystal form A of carbamate fumarate;

[0031] Figure 4 shows the amorphous X-ray powder diffraction (XRPD) pattern of urethane fumarate. Detailed Implementation

[0032] The present invention will be further described in detail below with reference to 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 made in the art based on the content disclosed in the present invention shall fall within the protection scope of the present invention.

[0033] The structure of the compound is determined by nuclear magnetic resonance (NMR). 1 Determined by ¹H NMR or LC-MS.

[0034] The liquid chromatography-mass spectrometry (LC-MS) system is an Agilent G6120B (compatible with an Agilent 1260 liquid chromatography system); the nuclear magnetic resonance spectrometer (… 1 H NMR) is Brukeravance ~400 or Brukeravance ~600, nuclear magnetic resonance (NMR) 1 ¹H NMR shifts (δ) are given in parts per million (ppm), the solvent is DMSO, the internal standard is tetramethylsilane (TMS), and the chemical shifts are expressed in 10⁻¹⁰ ppm. ～6 (ppm) is given as the unit.

[0035] In this invention, the term "room temperature" refers to a temperature between 10 and 30°C.

[0036] Comparative Example 1: Synthesis of (S)-5-(1-(2-chlorophenyl)-2-methoxy-2-oxoethyl-4,5,6,7-tetrahydrothiopheno[3,2-c]pyridin-2-yl-4-methylpiperazine-1-carboxylic acid ester)

[0037] A free alkali-yellow oily substance was prepared according to the preparation method of Example 1 in patent CN116262757A. After purification by column chromatography, the solid of the compound of Comparative Example 1 was obtained with a purity of 98.89%.

[0038] LC-MS(ESI+): m / z=464.2(M+H) + .

[0039] 1HNMR(400MHz, 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),3.67(s,3H),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).

[0040] Comparative Example 2: Synthesis of (S)-5-(1-(2-chlorophenyl)-2-methoxy-2-oxoethyl-4,5,6,7-tetrahydrothiopheno[3,2-c]pyridin-2-yl-4-methylpiperazine-1-carboxylate difumarate)

[0041] Using the compound obtained in Comparative Example 1 as the starting material, two molecules of fumarate solid with a purity of 99.38% were prepared according to the preparation method in Example 2 of Patent CN116262757A.

[0042] LC-MS(ESI+): m / z=464.2(M+H) + .

[0043] 1 HNMR(400MHz, DMSO-d6)δ:13.82~9.71(br,4H),7.64~7.56(m,1H),7.56~7.47(m,1H),7.46~7.35(m,2H),6.62(s,4H),6.3 9(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).

[0044] Example 1: Preparation of (S)-5-(1-(2-chlorophenyl)-2-methoxy-2-oxoethyl-4,5,6,7-tetrahydrothiopheno[3,2-c]pyridin-2-yl-4-methylpiperazine-1-carboxylic acid ester fumarate (crystal form A)

[0045] The synthesis route is as follows:

[0046] 5 ml of anhydrous ethanol and fumaric acid (0.70 g, 6 mmol) were added to a three-necked flask, and the mixture was heated and stirred until dissolved. Then, (S)-5-(1-(2-chlorophenyl)-2-methoxy-2-oxoethyl-4,5,6,7-tetrahydrothieno[3,2-c]pyridin-2-yl 4-methylpiperazine-1-carboxylate (1.86 g, 4 mmol) was added. The mixture was then cooled to room temperature and stirred for 1 h, resulting in the precipitation of a large amount of solid. The solid was filtered, dried, and yielded 1.74 g of crystalline (S)-5-(1-(2-chlorophenyl)-2-methoxy-2-oxoethyl-4,5,6,7-tetrahydrothieno[3,2-c]pyridin-2-yl 4-methylpiperazine-1-carboxylate fumarate, with a yield of 75% and a purity of 99.22%.

[0047] ESI-MS: m / z = 464.2(M+H) + .

[0048] 1 H NMR (400MHz, DMSO-d6) δ: 10.37 (brs, 2H), 7.58 (dd, 1H), 7.49 (dd, 1H), 7.39 (td, 1H), 7.36 (td, 1H), 6.62 (s, 2H), 6.36 (s, 1H), 4.84 (s, 1 H), 3.65(s, 3H), 3.52-3.58(m, 2H), 3.52(brs, 2H), 3.42-3.48(m, 2H), 2.74-2.87(m, 2H), 2.66(t, 2H), 2.45-2.51(m, 4H), 2.29(s, 3H).

[0049] The X-ray powder diffraction, differential scanning calorimetry, and thermogravimetric analysis results of (S)-5-(1-(2-chlorophenyl)-2-methoxy-2-oxoethyl-4,5,6,7-tetrahydrothiopheno[3,2-c]pyridin-2-yl-4-methylpiperazine-1-carboxylate fumarate crystal form A in this embodiment are shown in Figures 1 to 3.

[0050] Example 2: Preparation of (S)-5-(1-(2-chlorophenyl)-2-methoxy-2-oxoethyl-4,5,6,7-tetrahydrothiopheno[3,2-c]pyridin-2-yl-4-methylpiperazine-1-carboxylate fumarate (crystal form A)

[0051] 5 ml of anhydrous methanol and fumaric acid (0.70 g, 6 mmol) were added to a three-necked flask, and the mixture was heated and stirred until dissolved. Then, (S)-5-(1-(2-chlorophenyl)-2-methoxy-2-oxoethyl-4,5,6,7-tetrahydrothieno[3,2-c]pyridin-2-yl 4-methylpiperazine-1-carboxylate (1.86 g, 4 mmol) was added. The mixture was then cooled to room temperature and stirred for 1 h, resulting in the precipitation of a large amount of solid. The solid was filtered, dried, and yielded 1.39 g of crystalline (S)-5-(1-(2-chlorophenyl)-2-methoxy-2-oxoethyl-4,5,6,7-tetrahydrothieno[3,2-c]pyridin-2-yl 4-methylpiperazine-1-carboxylate fumarate, with a yield of 60% and a purity of 99.34%.

[0052] The mass spectrometry, proton spectrum, X-ray powder diffraction, differential scanning calorimetry, and thermogravimetric analysis of this embodiment are the same as those of Example 1, proving that it is the same crystal form.

[0053] Example 3: Preparation of (S)-5-(1-(2-chlorophenyl)-2-methoxy-2-oxoethyl-4,5,6,7-tetrahydrothiopheno[3,2-c]pyridin-2-yl-4-methylpiperazine-1-carboxylate fumarate (crystal form A)

[0054] 5 ml of isopropanol and fumaric acid (0.70 g, 6 mmol) were added to a three-necked flask, and the mixture was heated and stirred until dissolved. Then, (S)-5-(1-(2-chlorophenyl)-2-methoxy-2-oxoethyl-4,5,6,7-tetrahydrothieno[3,2-c]pyridin-2-yl 4-methylpiperazine-1-carboxylic acid ester (1.86 g, 4 mmol) was added. The mixture was then cooled to room temperature and stirred for 1 h, resulting in the precipitation of a large amount of solid. The solid was filtered, dried, and yielded 1.90 g of crystalline (S)-5-(1-(2-chlorophenyl)-2-methoxy-2-oxoethyl-4,5,6,7-tetrahydrothieno[3,2-c]pyridin-2-yl 4-methylpiperazine-1-carboxylic acid ester fumarate, with a yield of 82% and a purity of 98.55%.

[0055] The mass spectrometry, proton spectrum, X-ray powder diffraction, differential scanning calorimetry, and thermogravimetric analysis of this embodiment are the same as those of Example 1, proving that it is the same crystal form.

[0056] Example 4: Preparation of (S)-5-(1-(2-chlorophenyl)-2-methoxy-2-oxoethyl-4,5,6,7-tetrahydrothiopheno[3,2-c]pyridin-2-yl-4-methylpiperazine-1-carboxylate fumarate (crystal form A)

[0057] 10 ml of acetonitrile and fumaric acid (0.70 g, 6 mmol) were added to a three-necked flask, and the mixture was heated and stirred until dissolved. Then, (S)-5-(1-(2-chlorophenyl)-2-methoxy-2-oxoethyl-4,5,6,7-tetrahydrothieno[3,2-c]pyridin-2-yl 4-methylpiperazine-1-carboxylate (1.86 g, 4 mmol) was added. The mixture was then cooled to room temperature and stirred for 1 h, resulting in the precipitation of a large amount of solid. The solid was filtered, dried, and yielded 1.69 g of crystalline (S)-5-(1-(2-chlorophenyl)-2-methoxy-2-oxoethyl-4,5,6,7-tetrahydrothieno[3,2-c]pyridin-2-yl 4-methylpiperazine-1-carboxylate fumarate, with a yield of 73% and a purity of 98.87%.

[0058] The mass spectrometry, proton spectrum, X-ray powder diffraction, differential scanning calorimetry, and thermogravimetric analysis of this embodiment are the same as those of Example 1, proving that it is the same crystal form.

[0059] Example 5: Preparation of (S)-5-(1-(2-chlorophenyl)-2-methoxy-2-oxoethyl-4,5,6,7-tetrahydrothiopheno[3,2-c]pyridin-2-yl-4-methylpiperazine-1-carboxylate fumarate (amorphous)

[0060] 1.50 g of (S)-5-(1-(2-chlorophenyl)-2-methoxy-2-oxoethyl-4,5,6,7-tetrahydrothieno[3,2-c]pyridin-2-yl 4-methylpiperazine-1-carboxylate fumarate obtained by the preparation method in Example 1 was added to 75 ml of water, stirred at room temperature, filtered to remove insoluble matter, filtered under vacuum, and the filtrate was freeze-dried to obtain 1.32 g of white solid, with a yield of 88% and a purity of 99.17%.

[0061] The mass spectrometry and proton spectrum of this embodiment are the same as those of Example 1, and the X-ray powder diffraction detection results are shown in Figure 4.

[0062] Experimental Example 1: Stability Study

[0063] 1. Test sample

[0064] Comparative Example 1, Comparative Example 2, Examples 1 to 5.

[0065] 2. Test Methods

[0066] Three samples were weighed and placed in weighing bottles. Each sample group was placed under high temperature (60℃), high humidity (RH 92.5%), and light (5000 Lux) conditions. Samples were taken and tested after 10 days and 30 days, respectively.

[0067] Detection method: Accurately weigh appropriate amounts of each sample, dissolve and quantitatively dilute with acetonitrile to a concentration of approximately 0.1 mg / mL. High-performance liquid chromatography (HPLC) is used with octadecylsilane-bonded silica gel as the stationary phase. The mobile phase consists of a 40:60 volume ratio of 30 mmol / L ammonium acetate (adjusted to pH 5.0 with acetic acid) and acetonitrile (B). The detection wavelength is 220 nm, the flow rate is 1.0 mL / min, the column temperature is 30 °C, and the injection volume is 10 μL. Each sample solution is injected into the HPLC system, and the chromatograms are recorded. The content of the main component and impurities is calculated using the peak area normalization method.

[0068] 3. Test Results

[0069] Table 1. Results of the experiment on influencing factors

[0070] The experimental results are shown in the table above: After 30 days of testing under the influence of high temperature, high humidity and light, the stability of the samples in each embodiment was better than that of the comparative sample, proving that the carbamate monofumarate of the present invention and its crystalline and amorphous forms are more stable and more suitable for pharmaceutical use.

[0071] Experimental Example 2: Solubility Study

[0072] 1. Test sample

[0073] Comparative Example 1, Example 1, and Example 5.

[0074] 2. Test Methods

[0075] Each sample was accurately weighed, and 1 ml of physiological saline and pH 1.2 buffer solution were taken at 25–30°C. Then, 10 mg of each compound was added to each sample. The solubility data are shown in the table below.

[0076] Table 2 Solubility Test Results

[0077] The results showed that the compounds in the embodiments of the present invention have excellent solubility, especially meeting the clinical requirements for preparing effective concentrations of injection solutions, thus broadening the administration methods; while the solubility conditions of Comparative Example 1 are limited, and the requirements for administration methods and types of excipients are relatively high.

[0078] Experimental Example 3: Hygroscopicity Study

[0079] 1. Test sample

[0080] Comparative Example 1, Comparative Example 2, Example 1 and Example 5.

[0081] 2. Test Methods

[0082] (1) Take a dry glass weighing bottle with a stopper (outer diameter 50 mm, height 15 mm) and place it in a suitable constant temperature desiccator (with ammonium chloride or ammonium sulfate saturated solution placed at the bottom) or artificial climate chamber (temperature set at 25℃±1℃, relative humidity at 80%±2%) one day before the test, and accurately weigh it with the lid on (m1).

[0083] (2) Take an appropriate amount of the test sample and spread it evenly in the weighing bottle mentioned above. The thickness of the test sample is generally about 1 mm. Weigh it accurately with the cap on (m2).

[0084] (3) Leave the weighing bottle open and place it together with the bottle cap under the above constant temperature and humidity conditions for 24 hours;

[0085] (4) Close the weighing bottle cap and accurately weigh the weight (m3);

[0086] (5) Percentage gain = (m3-m2) / (m2-m1)×100%.

[0087] 3. Test Results

[0088] The hygroscopicity results are shown in the table below: The crystalline form A of the carbamate fumarate of the present invention (see Example 1) has no or almost no hygroscopicity; the amorphous form of the carbamate fumarate (see Example 5) has slight hygroscopicity; the free base of Comparative Example 1 has no or almost no hygroscopicity; and the difumarate of Comparative Example 2 has slight hygroscopicity. However, the weight gain percentage of all these forms is higher than that of crystalline form A and the amorphous form of the present invention, demonstrating that the crystalline and amorphous forms of the carbamate fumarate of the present invention have better hygroscopicity characteristics and are significantly superior to the difumarate of Comparative Example 2. Therefore, the carbamate fumarate of the present invention, as well as its crystalline form A and amorphous form, are more advantageous for product storage and industrial application.

[0089] Table 3. Results of Hygroscopicity

[0090] According to the pharmacopoeia definition, the above results are as follows: "hygroscopic" means that the weight gain due to hygroscopic absorption is less than 15% but not less than 2%; "extremely hygroscopic" means that the weight gain due to hygroscopic absorption is not less than 15%; "no or almost no hygroscopicity" means that the weight gain due to hygroscopic absorption is less than 0.2%; "slightly hygroscopic" means that the weight gain due to hygroscopic absorption is less than 2% but not less than 0.2%.

[0091] Experimental Example 4: Study on Antiplatelet Aggregation Effect

[0092] 1. Purpose of the experiment: To evaluate and compare the antiplatelet aggregation effects of various compounds after equimolar administration.

[0093] 2. Experimental Samples:

[0094] Comparative Example 1, Comparative Example 2, Example 1, Example 3 and Example 5 were prepared into gastric-soluble capsules by filling different gastric-soluble capsules in the same batch according to the dosage.

[0095] 3. Test methods

[0096] 3.1 Grouping and Administration: Twenty-four beagle dogs were randomly and evenly divided into six groups of four each: solvent control group, comparative example 1 group, comparative example 2 group, example 1 group, example 3 group, and example 5 group. Administration was performed in two cycles with a 14-day washout period. Each group received a single oral gavage of a gastric-soluble capsule at a dose of 6 μmol / kg. Blood samples were collected from the forelimb veins three hours after administration for testing.

[0097] 3.2 Sampling method: Each blood sample is approximately 1.8 mL, which is directly added to a glass vacuum tube containing sodium citrate anticoagulant. After blood collection, the tube should be gently inverted and mixed 5 to 10 times to ensure that the blood and anticoagulant are fully mixed.

[0098] 3.3 Sample preparation: (1) Platelet-rich plasma (PRP) preparation: Centrifuge 200g of sodium citrate anticoagulated blood sample at 25℃ for 10min, and take about 300μL for later use; (2) Platelet-poor plasma (PPP) preparation: Centrifuge 3000g of the blood sample with PRP taken above at 25℃ for 10min, and place at room temperature; (3) Platelet aggregation rate is measured using a platelet aggregator (Helena Laboratories, USA, model: AgG RAM): First, take the PPP corresponding to the PRP to be tested in each channel for transmittance correction. After correction, take out the PPP, and then put the cuvette with 225μl of the PRP to be tested into each channel, add a stir bar, and then add ADP (adenosine diphosphate, final concentration of 20μM), and immediately start the platelet maximum aggregation rate detection.

[0099] 4. Test Results

[0100] The maximum platelet aggregation rate data of each group after equimolar administration are shown in the table below. The results showed that the maximum platelet aggregation rate (%) in the solvent control group was 77.13±3.00; the maximum platelet aggregation rates (%) in Comparative Example 1, Comparative Example 2, Example 1, Example 3, and Example 5 were 30.40±5.80, 25.54±5.39, 17.59±3.29, 16.84±4.82, and 19.96±3.01, respectively. Compared with the solvent control group, all groups showed significant inhibition of ADP-induced platelet aggregation in beagle dogs (P<0.0001); compared with Comparative Example 2, Example 1, Example 3, and Example 5 also showed significant inhibition of ADP-induced platelet aggregation in beagle dogs (P<0.01). This demonstrates that the carbamate fumarate of the present invention, and its crystalline and amorphous forms, have superior antiplatelet aggregation effects and better efficacy.

[0101] Table 4. Results of antiplatelet aggregation efficacy experiments in each group after equimolar administration.

[0102] Note: Compared with the solvent control group, **** indicates P<0.0001; compared with comparative group 2, ▲▲ indicates P<0.01.

[0103] The above embodiments are merely one of the preferred embodiments of the present invention and should not be used to limit the scope of protection of the present invention. Any modifications or refinements made to the main design concept and spirit of the present invention that are not of substantial significance, but solve the same technical problem as the present invention, should be included within the scope of protection of the present invention.

Claims

1. A carbamate fumarate as shown in formula (1):

2. A crystalline form A of the carbamate fumarate of claim 1, wherein the X-ray powder diffraction pattern of the crystalline form, when subjected to Cu-Kα radiation, exhibits diffraction peaks at the following 2θ values: 3.58±0.2°, 7.17±0.2°, 10.38±0.2°, 14.04±0.2°, 16.75±0.2°, 17.84±0.2°, and 18.49±0.2°.

3. The crystal form A according to claim 2, wherein the X-ray powder diffraction pattern of the crystal form further includes any one or more diffraction peaks with 2θ values ​​as follows: 11.82±0.2°, 12.98±0.2°, 22.96±0.2°, 23.21±0.2°, 23.45±0.2°, 25.27±0.2°.

4. The crystalline Form A of claim 2, having an X-ray powder diffraction pattern comprising the following 2Θ values and relative intensities of the diffraction peaks:

5. The crystalline Form A of claim 4, further comprising X-ray powder diffraction pattern of said crystalline Form A comprising one or more of the following sets of 2θ values and relative intensities of the diffraction peaks:

6. The crystal form A according to claim 2, wherein the crystal form has an X-ray powder diffraction pattern substantially as shown in Figure 1.

7. According to claim 2, the differential scanning calorimetry (DSC) spectrum of the crystal form has an endothermic peak at 150℃±5℃; preferably, the crystal form has a differential scanning calorimetry curve as shown in Figure 2.

8. Crystal form A according to claim 2, wherein the crystal form has a thermogravimetric analysis spectrum as shown in Figure 3.

9. An amorphous form of the carbamate fumarate of claim 1, wherein the X-ray powder diffraction pattern of the amorphous form, when subjected to Cu-Kα radiation, shows diffuse peaks; preferably, the amorphous form has an X-ray powder diffraction pattern substantially as shown in Figure 4.

10. A method for preparing the carbamate fumarate of claim 1 or the crystal form A of any one of claims 2-8, comprising the following steps: (S)-5-(1-(2-chlorophenyl)-2-methoxy-2-oxoethyl-4,5,6,7-tetrahydrothieno[3,2-c]pyridin-2-yl-4-methylpiperazine-1-carboxylic acid ester reacting with fumaric acid under protic solvent conditions to form a salt; after the reaction is complete, the temperature is lowered, a solid precipitates, and the solid is filtered and dried to obtain carbamate fumarate crystals; wherein the protic solvent is selected from alcohols or acetonitrile; preferably, the alcohol is selected from methanol, ethanol or isopropanol.

11. A method for preparing the amorphous solid according to claim 9, comprising the following steps: adding crystal A according to any one of claims 2-8 to a solvent, filtering after the solid dissolves, and spray-drying or freeze-drying the filtrate to obtain a white solid; wherein the solvent is selected from tert-butanol or water.

12. A pharmaceutical composition comprising the carbamate fumarate of claim 1, crystal form A of any one of claims 2-8 or the amorphous form of claim 9, and further comprising a pharmaceutically acceptable carrier.

13. A pharmaceutical formulation comprising the carbamate fumarate of claim 1, crystal form A of any one of claims 2-8 or the amorphous form of claim 9, wherein the pharmaceutical formulation is selected from oral formulations or liquid formulations; preferably, the liquid formulation comprises an injection or a lyophilized formulation.

14. Use of the carbamate fumarate of claim 1, crystal form A of any one of claims 2-8, amorphous form of claim 9, pharmaceutical composition of claim 12, or pharmaceutical formulation of claim 13 in the preparation of a medicament for the prevention and / or treatment of cardiovascular, cerebrovascular, and other circulatory disorders caused by platelet aggregation; preferably, the cardiovascular, cerebrovascular, and other circulatory disorders caused by platelet aggregation include, but are not limited to, acute coronary syndrome, atherosclerotic disease, or thrombotic complications.

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