Crystal form iii of oxazolidinone derivative, its preparation method and use

Abstract

The application belongs to the technical field of medicines, and relates to a crystal form III of an oxazolidinone derivative, a preparation method and purposes of the crystal form III, in particular to a crystal form III of (S)-5-chloro-N-((2-oxo-3-(4-(2-oxo-2H-pyridin-1-yl)phenyl)-1,3-oxazolidin-5-yl)methyl)thiophene-2-formamide, a preparation method and purposes of the crystal form III.

Description

Technical Field

[0001] This invention belongs to the field of pharmaceutical technology and relates to a crystal form III of an oxazolidinone derivative, its preparation method and uses. Specifically, this invention relates to a crystal form III of (S)-5-chloro-N-((2-oxo-3-(4-(2-oxo-2H-pyridin-1-yl)phenyl)-1,3-oxazolidin-5-yl)methyl)thiophene-2-carboxamide, its preparation method and uses. Background Technology

[0002] "Shaban" drugs are a new generation of antithrombotic agents, primarily used to prevent and treat acute and severe cardiovascular and cerebrovascular diseases such as venous thrombosis, pulmonary embolism, and systemic embolism caused by atrial fibrillation. Shaban anticoagulants play a crucial role in the treatment, prevention, and management of thromboembolic diseases. This class of drugs overcomes the shortcomings of traditional drugs such as heparin and warfarin, and has become a model of a new type of orally administered anticoagulant. Currently, only a few shaban drugs are available globally, and all are controlled by multinational corporations.

[0003] (S)-5-chloro-N-((2-oxo-3-(4-(2-oxo-2H-pyridin-1-yl)phenyl)-1,3-oxazolidin-5-yl)methyl)thiophene-2-carboxamide (Compound I) is a domestically developed innovative drug, the first domestically developed anticoagulant drug of the xaban class to enter the clinical research stage. Its structure is as follows:

[0004]

[0005] The study of drug polymorphs has become increasingly important. Different polymorphs of a drug substance can have different chemical and physical properties, including melting point, chemical reactivity, apparent solubility, dissolution rate, optical and mechanical properties, vapor pressure, and density. These properties can directly affect the processing and / or production of drug substances and formulations, and can also affect the stability, solubility, and bioavailability of formulations. Therefore, polymorphism can affect the quality, safety, and efficacy of drug formulations. Currently reported polymorphs of compound I include polymorph I and polymorph II. Patent CN201310183080.4 discloses polymorph I of compound I, prepared using a mixed solution of formic acid and ethanol as a solvent, through dissolution and crystallization to obtain polymorph I. The characteristic absorption peak (2θ) values ​​of this polymorph in X-ray powder diffraction are: 4.04, 8.08, 12.16, 13.96, 17.14, 19.40, 19.76, 20.30, 23.80, 24.46, 28.60, 32.26; the 2θ measurement error is... ±0.2; Patent CN201310182806.2 discloses crystal form II of compound I. The preparation method uses ethanol or water as a solvent to perform dissolution and crystallization to obtain crystal form II. The characteristic absorption peak (2θ) values ​​of this crystal form in X-ray powder diffraction are: 6.96, 9.68, 13.46, 13.94, 15.16, 15.50, 19.42, 20.60, 21.44, 22.72, 23.20, 24.66, 25.72, 27.32, 27.78; the 2θ measurement error is ±0.2.

[0006] The literature “Liu Wei; Yuan Jing; Zhang Shijun et al. Synthesis, crystal structure and anticoagulant activity of 5-chloro-N-[[(5S)-2-oxo-3-[4-(2-oxopyridin-1(2H)-yl)phenyl]oxazolidin-5-yl]methyl]thiophene-2-carboxamide[J]. Structural Chemistry. 2014, 33(7):1091-1095” discloses the preparation method of compound I and its single crystal, but does not disclose its crystal form data. Based on the preparation method of compound I disclosed in the literature, the experimental process was repeated to obtain crystal form I and crystal form II of compound I, as shown in Comparative Examples 3 and 4. Patent CN201410209934.6 also discloses a method for preparing a solvate of compound I, acetic acid. This method uses acetic acid as a solvent for dissolution and crystallization. The characteristic absorption peaks (2θ) of its crystal form in the X-ray powder diffraction pattern are: 3.60, 7.04, 10.50, 14.00, 17.48, 20.12, 21.00, 23.86, 24.56, 26.42, 28.12; the 2θ measurement error is ±0.2. Additionally, patent CN201110337461.4 also reports a method for preparing purified acetic acid compound I. Compound I has a melting point of 204.8-205.8°C, and the final product was found to contain trace amounts of acetic acid.

[0007] Although crystal forms I and II of compound I have certain therapeutic effects in the treatment, prevention and treatment of thromboembolic diseases, the solid form of crystal form I of compound I is unstable and will transform into crystal form II when placed under high temperature, high humidity or light conditions. Crystal form II has low apparent solubility and low bioavailability, so the use of the above two crystal forms for oral solid dosage form development is greatly limited. Therefore, the research on new crystal forms of compound I is very necessary. Summary of the Invention

[0008] In view of the above, the present invention provides a crystal form III of an oxazolidinone derivative, a method for its preparation, and its uses, in order to at least partially solve at least one of the aforementioned technical problems.

[0009] As one aspect of the present invention, a crystal form III of (S)-5-chloro-N-((2-oxo-3-(4-(2-oxo-2H-pyridin-1-yl)phenyl)-1,3-oxazolidine-5-yl)methyl)thiophene-2-carboxamide (compound I) is provided, wherein the X-ray powder diffraction of the crystal form III exhibits characteristic peaks at diffraction angles 2θ = 5.76 ± 0.2°, 11.68 ± 0.2°, 14.98 ± 0.2°, 16.38 ± 0.2°, and 16.96 ± 0.2°.

[0010] Preferably, the X-ray powder diffraction of the crystal form III exhibits characteristic peaks at diffraction angles 2θ = 5.76 ± 0.2°, 8.30 ± 0.2°, 9.16 ± 0.2°, 11.68 ± 0.2°, 14.98 ± 0.2°, 16.38 ± 0.2°, and 16.96 ± 0.2°.

[0011] Preferably, the (S)-5-chloro-N-((2-oxo-3-(4-(2-oxo-2H-pyridin-1-yl)phenyl)-1,3-oxazolidine-5-yl)methyl)thiophene-2-carboxamide (compound I) in crystal form III is characterized by X-ray powder diffraction of crystal form III at diffraction angles 2θ = 5.76 ± 0.2°, 8.30 ± 0.2°, 9.16 ± 0.2°, 11.68 ± 0.2°, 14.98 ± 0.2°, 16.38 ± 0.2°, 16.96 ± 0.2°, 21.22 ± 0.2°, 23.40 ± 0.2°, 25.20 ± 0.2°, and 26.70 ± 0.2°.

[0012] Preferably, the (S)-5-chloro-N-((2-oxo-3-(4-(2-oxo-2H-pyridin-1-yl)phenyl)-1,3-oxazolidine-5-yl)methyl)thiophene-2-carboxamide (compound I) is in crystal form III, and the X-ray powder diffraction of crystal form III is at diffraction angles 2θ = 5.76 ± 0.2°, 8.30 ± 0.2°, and 9.16 ± Characteristic peaks are observed at 0.2°, 11.68±0.2°, 14.98±0.2°, 16.38±0.2°, 16.96±0.2°, 21.22±0.2°, 23.40±0.2°, 23.98±0.2°, 25.20±0.2°, 26.70±0.2°, 30.22±0.2°, and 39.78±0.2°.

[0013] As a second aspect of the present invention, a method for preparing crystal form III of (S)-5-chloro-N-((2-oxo-3-(4-(2-oxo-2H-pyridin-1-yl)phenyl)-1,3-oxazolidine-5-yl)methyl)thiophene-2-carboxamide (compound I) is provided, comprising the following steps:

[0014] (1) Dissolve compound I in a suitable solvent to prepare compound I solution, connect compound I solution to a horizontal flow pump, and control the working pressure to 7.5-30 MPa;

[0015] (2) The carbon dioxide in the steel cylinder is fed into the supercritical fluid antisolvent equipment system through a constant speed and constant pressure pump and enters the crystallization kettle. The flow rate is controlled at 10-40 mL / min, the start-up temperature is controlled at 33-60℃, and the working pressure is controlled at 7.5-30 MPa.

[0016] (3) The compound I solution prepared in step (1) above is rapidly injected into the crystallization vessel through the nozzle in the supercritical fluid antisolvent equipment system by a horizontal flow pump. The flow rate is controlled at 0.05-5 mL / min. After the compound I solution is added, carbon dioxide is continuously passed through for drying.

[0017] (4) Collect the crystals of compound I that have precipitated from the solvent at the bottom of the crystallization vessel. The resulting product is the crystal form III of compound I.

[0018] Preferably, in the method for preparing (S)-5-chloro-N-((2-oxo-3-(4-(2-oxo-2H-pyridin-1-yl)phenyl)-1,3-oxazolidine-5-yl)methyl)thiophene-2-carboxamide (compound I) to crystal form III, the mass-to-volume ratio of compound I to solvent in step (1) is 1:15-1:50 g / mL, preferably 1:28-1:35 g / mL.

[0019] Preferably, in the method for preparing (S)-5-chloro-N-((2-oxo-3-(4-(2-oxo-2H-pyridin-1-yl)phenyl)-1,3-oxazolidine-5-yl)methyl)thiophene-2-carboxamide (compound I) in crystal form III, the solvent in step (1) is selected from one or more of dimethylformamide, dimethyl sulfoxide, methanol, ethanol, acetone, dichloromethane, and tetrahydrofuran.

[0020] Preferably, in the method for preparing (S)-5-chloro-N-((2-oxo-3-(4-(2-oxo-2H-pyridin-1-yl)phenyl)-1,3-oxazolidine-5-yl)methyl)thiophene-2-carboxamide (compound I) to crystal form III, the solvent in step (1) is selected from dimethylformamide or dimethyl sulfoxide.

[0021] Preferably, in the method for preparing (S)-5-chloro-N-((2-oxo-3-(4-(2-oxo-2H-pyridin-1-yl)phenyl)-1,3-oxazolidin-5-yl)methyl)thiophene-2-carboxamide (compound I) crystal form III, the working pressure in steps (1) and (2) is selected from 10 to 13 MPa, and the starting temperature in step (2) is selected from 40 to 50 °C.

[0022] Preferably, in the method for preparing (S)-5-chloro-N-((2-oxo-3-(4-(2-oxo-2H-pyridin-1-yl)phenyl)-1,3-oxazolidine-5-yl)methyl)thiophene-2-carboxamide (compound I) crystal form III, the flow rate in step (2) is selected from 15 to 25 mL / min, and the flow rate in step (3) is selected from 0.5 to 1 mL / min.

[0023] Preferably, the use of (S)-5-chloro-N-((2-oxo-3-(4-(2-oxo-2H-pyridin-1-yl)phenyl)-1,3-oxazolidin-5-yl)methyl)thiophene-2-carboxamide (compound I) crystal form III in the prevention / treatment of thromboembolic diseases and / or thromboembolic complications.

[0024] Preferably, the use of (S)-5-chloro-N-((2-oxo-3-(4-(2-oxo-2H-pyridin-1-yl)phenyl)-1,3-oxazolidin-5-yl)methyl)thiophene-2-carboxamide (compound I) crystal form III in an oral medication for the prevention / treatment of thromboembolic diseases and / or thromboembolic complications.

[0025] As a third aspect of the invention, the use of crystal form III of (S)-5-chloro-N-((2-oxo-3-(4-(2-oxo-2H-pyridin-1-yl)phenyl)-1,3-oxazolidin-5-yl)methyl)thiophene-2-carboxamide (compound I) in the preparation of a treatment, prevention and treatment of thromboembolic diseases is provided.

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

[0027] (1) The crystal form III of compound I provided by the present invention has good stability as shown by the influencing factor test, accelerated test and long-term stability test of room temperature sample retention. The dissolution rate test shows that the crystal form III of compound I has a faster dissolution rate under both acidic and neutral conditions, which is more conducive to improving its dissolution rate and absorption in vivo, thereby improving its bioavailability and efficacy.

[0028] (2) The method for preparing crystal form III of compound I provided by the present invention uses a supercritical crystallization device and an antisolvent method for drug crystallization. The operation is simple, the equipment has low energy consumption and low cost. The obtained crystal form III of compound I has good stability and a fast dissolution rate, which is more conducive to absorption in vivo, thereby improving its bioavailability and efficacy.

[0029] (3) The present invention provides the use of the preparation of crystal form III of (S)-5-chloro-N-((2-oxo-3-(4-(2-oxo-2H-pyridin-1-yl)phenyl)-1,3-oxazolidin-5-yl)methyl)thiophene-2-carboxamide (compound I) in the preparation of treatment, prevention and treatment of thromboembolic diseases.

[0030] Compared with the prior art, the present invention has the following beneficial effects:

[0031] The crystal form III of compound I provided by this invention, as determined by influencing factor tests, accelerated stability tests, and long-term stability tests at room temperature, demonstrates good stability. Dissolution rate tests show that crystal form III of compound I dissolves significantly faster than crystal forms I and II. Therefore, it is more conducive to improving its dissolution rate and absorption in vivo, thereby enhancing its bioavailability and efficacy. Attached Figure Description

[0032] Figure 1 This is the X-ray powder diffraction pattern of crystal form III of compound I obtained in Example 1 of the invention.

[0033] Figure 2 This is the X-ray powder diffraction pattern of crystal form I of compound I obtained in Comparative Example 1;

[0034] Figure 3 This is the X-ray powder diffraction pattern of crystal form II of compound I obtained in Comparative Example 2;

[0035] Figure 4 This is a schematic diagram of the connection of the supercritical fluid antisolvent equipment in the preparation process of crystal form III of compound I involved in this invention. Detailed Implementation

[0036] The present invention will be further described below through embodiments, but these descriptions are not intended to further limit the scope of the invention. Those skilled in the art should understand that equivalent substitutions or corresponding improvements made to the technical features of the present invention still fall within the protection scope of the present invention.

[0037] In the following examples, the same reagents and reagents were from the same batch number, and compound I was prepared in-house.

[0038] The powder diffraction instrument used in this invention is a Rigaku D / max-2500 powder diffractometer, a product of Rigaku Corporation, Japan.

[0039] The supercritical crystallization equipment used in this invention is the SCF-PD-200 type supercritical crystallization equipment.

[0040] During the study of the crystal form of compound I, the inventors discovered a novel crystal form of compound I, which they named crystal form III. Crystal form III of compound I exhibits excellent stability. This invention employs an SCF-PD-200 supercritical crystallization apparatus and utilizes an antisolvent method for drug crystallization. This involves dissolving the drug in a suitable solvent and then mixing it with supercritical carbon dioxide. The property of carbon dioxide—compatibility with the solvent but insolubility for the drug—allows for rapid crystallization. First, the solvent system is determined based on the solvent's solubility, polarity, and viscosity. The phase conditions of carbon dioxide (temperature, pressure) and other process parameters (drug concentration, flow rate, carbon dioxide flow rate, etc.) are optimized to obtain crystal form III of compound I.

[0041] Invention Embodiments

[0042] Example 1: Preparation of Crystal Form III of Compound I

[0043] Example 1-1

[0044] (1) Dissolve 10g of compound I in 330mL of dimethylformamide to prepare compound I solution. Connect compound I solution to a horizontal flow pump and control the working pressure to 12MPa.

[0045] (2) The carbon dioxide in the cylinder is fed into the supercritical fluid antisolvent equipment system through a constant speed and constant pressure pump and enters the crystallization kettle. The flow rate is controlled at 20 mL / min, the start-up temperature is controlled at 45℃, and the working pressure is controlled at 12 MPa.

[0046] (3) The compound I solution prepared in step (1) above is rapidly injected into the crystallization vessel through the nozzle in the supercritical fluid antisolvent equipment system by a horizontal flow pump. The flow rate is controlled at 0.5 mL / min. After the compound I solution is added, carbon dioxide is continuously passed through for drying.

[0047] (4) Collect the crystals of compound I that have precipitated from the solvent at the bottom of the crystallization vessel. The resulting product is the crystal form III of compound I.

[0048] X-ray powder diffraction was performed on the dried crystals. Characteristic peaks were observed at 2θ = 5.76°, 8.30°, 9.16°, 11.68°, 14.98°, 16.38°, 16.96°, 21.22°, 22.44°, 23.40°, 23.98°, 25.20°, 26.70°, 30.22°, and 39.78°, as detailed in the attached instruction manual. Figure 1 As shown, its melting point is 200℃.

[0049] Examples 1-2

[0050] (1) Dissolve 10g of compound I in 280mL of dimethyl sulfoxide to prepare compound I solution. Connect the compound I solution to a horizontal flow pump and control the working pressure to 10MPa.

[0051] (2) The carbon dioxide in the cylinder is fed into the supercritical fluid antisolvent equipment system through a constant speed and constant pressure pump and enters the crystallization kettle. The flow rate is controlled at 15 mL / min, the start-up temperature is controlled at 45℃, and the working pressure is controlled at 10 MPa.

[0052] (3) The compound I solution prepared in step (1) is rapidly injected into the crystallization vessel through a nozzle in the supercritical fluid antisolvent equipment system using a horizontal flow pump. The flow rate is controlled at 1 mL / min. After the compound I solution is added, carbon dioxide is continuously passed through for drying.

[0053] (4) The crystals of compound I precipitated from the solvent were collected at the bottom of the crystallization vessel. The resulting product is the crystal form III of compound I. X-ray powder diffraction was measured on the dried crystals. The X-ray powder diffraction showed characteristic peaks at 2θ = 5.76°, 8.29°, 9.16°, 11.68°, 14.99°, 16.38°, 16.96°, 21.22°, 22.44°, 23.40°, 23.97°, 25.20°, 26.70°, 30.22°, and 39.78°.

[0054] Examples 1-3

[0055] (1) Dissolve 10g of compound I in 350mL of methanol to prepare compound I solution. Connect compound I solution to a horizontal flow pump and control the working pressure to 13MPa.

[0056] (2) The carbon dioxide in the cylinder is fed into the supercritical fluid antisolvent equipment system through a constant speed and constant pressure pump and enters the crystallization kettle. The flow rate is controlled at 25 mL / min, the start-up temperature is controlled at 33℃, and the working pressure is controlled at 13 MPa.

[0057] (3) The compound I solution prepared in step (1) is rapidly injected into the crystallization vessel through a nozzle in the supercritical fluid antisolvent equipment system using a horizontal flow pump. The flow rate is controlled at 2 mL / min. After the compound I solution is added, carbon dioxide is continuously passed through for drying.

[0058] (4) The crystals of compound I precipitated from the solvent were collected at the bottom of the crystallization vessel. The resulting product is the crystal form III of compound I. X-ray powder diffraction was measured on the dried crystals. The X-ray powder diffraction showed characteristic peaks at 2θ = 5.78°, 8.30°, 9.16°, 11.68°, 14.98°, 16.38°, 16.96°, 21.22°, 22.45°, 23.40°, 23.98°, 25.20°, 26.69°, 30.22°, and 39.78°.

[0059] Examples 1-4

[0060] (1) Dissolve 10g of compound I in 150mL of acetone to prepare compound I solution. Connect compound I solution to a horizontal flow pump and control the working pressure to 7.5MPa.

[0061] (2) The carbon dioxide in the cylinder is fed into the supercritical fluid antisolvent equipment system through a constant speed and constant pressure pump and enters the crystallization kettle. The flow rate is controlled at 10 mL / min, the start-up temperature is controlled at 50℃, and the working pressure is controlled at 7.5 MPa.

[0062] (3) The compound I solution prepared in step (1) above is rapidly injected into the crystallization vessel through the nozzle in the supercritical fluid antisolvent equipment system by a horizontal flow pump. The flow rate is controlled at 0.05 mL / min. After the compound I solution is added, carbon dioxide is continuously passed through for drying.

[0063] (4) The crystals of compound I precipitated from the solvent were collected at the bottom of the crystallization vessel. The resulting product is the crystal form III of compound I. X-ray powder diffraction was measured on the dried crystals. The X-ray powder diffraction showed characteristic peaks at 2θ = 5.76°, 8.28°, 9.16°, 11.68°, 14.98°, 16.38°, 16.96°, 21.21°, 22.44°, 23.40°, 23.98°, 25.21°, 26.70°, 30.22°, and 39.78°.

[0064] Examples 1-5

[0065] (1) Dissolve 10g of compound I in 500mL of dichloromethane to prepare compound I solution. Connect compound I solution to a horizontal flow pump and control the working pressure to 15MPa.

[0066] (2) The carbon dioxide in the cylinder is fed into the supercritical fluid antisolvent equipment system through a constant speed and constant pressure pump and enters the crystallization kettle. The flow rate is controlled at 20 mL / min, the start-up temperature is controlled at 40℃, and the working pressure is controlled at 15 MPa.

[0067] (3) The compound I solution prepared in step (1) is rapidly injected into the crystallization vessel through a nozzle in the supercritical fluid antisolvent equipment system using a horizontal flow pump. The flow rate is controlled at 5 mL / min. After the compound I solution is added, carbon dioxide is continuously passed through for drying.

[0068] (4) Collect the crystals of compound I precipitated from the solvent at the bottom of the crystallization vessel. The resulting product is the crystal form III of compound I. X-ray powder diffraction was measured on the dried crystals. The X-ray powder diffraction showed characteristic peaks at 2θ = 5.75°, 8.30°, 9.17°, 11.68°, 14.98°, 16.38°, 16.96°, 21.22°, 22.44°, 23.41°, 23.98°, 25.20°, 26.70°, 30.22°, and 39.78°.

[0069] Examples 1-6

[0070] (1) Dissolve 10g of compound I in 400mL of tetrahydrofuran to prepare compound I solution. Connect compound I solution to a horizontal flow pump and control the working pressure to 30MPa.

[0071] (2) The carbon dioxide in the cylinder is fed into the supercritical fluid antisolvent equipment system through a constant speed and constant pressure pump and enters the crystallization kettle. The flow rate is controlled at 40 mL / min, the start-up temperature is controlled at 60℃, and the working pressure is controlled at 30 MPa.

[0072] (3) The compound I solution prepared in step (1) above is rapidly injected into the crystallization vessel through a nozzle in the supercritical fluid antisolvent equipment system using a horizontal flow pump. The flow rate is controlled at 0.3 mL / min. After the compound I solution is added, carbon dioxide is continuously passed through for drying.

[0073] (4) The crystals of compound I precipitated from the solvent were collected at the bottom of the crystallization vessel. The resulting product is the crystal form III of compound I. X-ray powder diffraction was measured on the dried crystals. The X-ray powder diffraction showed characteristic peaks at 2θ = 5.76°, 8.32°, 9.16°, 11.68°, 14.98°, 16.38°, 16.95°, 21.22°, 22.44°, 23.40°, 23.98°, 25.20°, 26.68°, 30.22°, and 39.78°.

[0074] It should be understood that the diffraction intensity of the characteristic peaks may vary slightly depending on the crystal preparation technique, sample mounting method, and measuring instrument, and this should also be within the scope of this invention. Furthermore, differences in instruments and other factors may affect the diffraction angle 2θ value, so the aforementioned diffraction angle 2θ value with characteristic peaks can vary within ±0.2° of the existing value.

[0075] Example 2: Stability test of crystal form III of compound I

[0076] HPLC analysis for the determination of related substances in compound I:

[0077] The HPLC chromatographic conditions were as follows: column: octadecylsilane bonded silica gel; mobile phase: acetonitrile-water (1:1); detection wavelength: 254 nm.

[0078] The relevant substances were determined using the normalization method.

[0079] Example 2-1 Influencing Factor Experiment

[0080] Take an appropriate amount of compound I sample, place it in a weighing bottle, spread it into a thin layer ≤5mm thick, and conduct the following experiment. The results are shown in Table 1.

[0081] l. High temperature test

[0082] The test sample was placed in a weighing bottle with the opening open and placed at 60℃ for 30 days. Samples were taken on days 0, 5, and 10 to detect the content and related substances, and samples were taken on days 0, 5, 10, and 30 to detect XRD data.

[0083] 2. High humidity test

[0084] Take an appropriate amount of the test sample and place it in a constant humidity desiccator. Place it at 25℃ and 92.5% relative humidity for 30 days. Take samples on days 0, 5, and 10 to detect the content and related substances. Take samples on days 0, 5, 10, and 30 to detect XRD data.

[0085] 3. Strong light irradiation test

[0086] The sample was placed open in a light box equipped with fluorescent lamps and kept under an illuminance of 4500 lx ± 500 lx for 30 days. Samples were taken on days 0, 5, and 10 to detect the content and related substances, and XRD data were collected on days 0, 5, 10, and 30.

[0087] Table 1. Experimental results of factors influencing compound I.

[0088]

[0089]

[0090] The above experimental results show that the content of crystal form III of compound I is stable under high temperature, high humidity, or strong light irradiation conditions, and its stability is comparable to that of crystal forms I and II. Crystal form I of compound I is unstable in its solid state and will transform into crystal form II when placed under high temperature, high humidity, or light irradiation conditions.

[0091] Example 2-2 Accelerated Test

[0092] Three batches of crystal form III of compound I were taken and placed at a temperature of 40℃±2℃ and a relative humidity of 75%±5% for 6 months. Samples were taken in the 1st, 2nd, 3rd and 6th months for testing, and the results are shown in Table 2.

[0093] Table 2. Accelerated test results for crystal form III of compound I.

[0094]

[0095]

[0096] Example 2-3 Long-term stability test of samples kept at room temperature

[0097] Long-term stability test of crystal form III of compound I at room temperature: Three batches of crystal form III of compound I were taken and placed at a temperature of 30℃±2℃ and a relative humidity of 60%±10% for 24 months. Samples were taken at 1, 2, 3, 6, 12, 18 and 24 months for testing. The results are shown in Table 3.

[0098] Table 3. Long-term stability test of crystal form III of compound I at room temperature.

[0099]

[0100]

[0101] The results of the influencing factors, accelerated testing and long-term stability test at room temperature for crystal form III of compound I showed that there were no significant changes in any of the test items for this crystal form, indicating good stability. In addition, X-ray powder diffraction test was also conducted, and the results showed that the crystal form did not change, and crystal form III of compound I can maintain good stability.

[0102] Invention Example 3: Dissolution Rate Test of Compound I

[0103] Example 3-1

[0104] Prepare three 100mL hydrochloric acid solutions with a pH of 1.2. Add 50mg of crystal form III, crystal form I, and crystal form II of compound I to each of the three prepared hydrochloric acid solutions, respectively. Shake for 24 hours using an air bath constant temperature shaker at 37℃ and a mains speed of 150rpm. Measure the maximum apparent solubility at different time points. The results are as follows:

[0105] Crystal form 0.5h 1h 2h 6h 24h Crystal form III 9.28 μg / mL 12.15 μg / mL 17.24 μg / mL 21.98 μg / mL 9.31 μg / mL Crystal form I 5.56 μg / mL 8.23 μg / mL 13.01 μg / mL 15.01 μg / mL 7.65 μg / mL Crystal form II 3.63 μg / mL 5.18 μg / mL 6.29 μg / mL 7.42 μg / mL 6.50 μg / mL

[0106] Example 3-2

[0107] Add 0.2 g of sodium dodecyl sulfate to 100 mL of hydrochloric acid solution with pH 1.2, stir to dissolve, and prepare 3 aliquots for later use. Add 50 mg of crystal form III, crystal form I, and crystal form II of compound I to the 3 prepared aliquots respectively. Shake for 24 hours using an air bath constant temperature shaker at 37℃ and 150 rpm. Measure the maximum apparent solubility at different time points. The results are as follows:

[0108]

[0109]

[0110] Example 3-3

[0111] Three 100mL phosphate buffer solutions with a pH of 6.8 were prepared. 50mg of crystal form III, crystal form I, and crystal form II of compound I were added to each of the three prepared phosphate buffer solutions, respectively. The solutions were shaken for 24 hours at 37℃ and 150rpm using an air bath constant-temperature shaker. The maximum apparent solubility was measured at different time points, and the results are as follows:

[0112] Crystal form 0.5h 1h 2h 6h 24h Crystal form III 10.69 μg / mL 14.13 μg / mL 19.46 μg / mL 23.56 μg / mL 11.64 μg / mL Crystal form I 6.46 μg / mL 9.32 μg / mL 17.96 μg / mL 19.06 μg / mL 9.43 μg / mL Crystal form II 4.33 μg / mL 5.53 μg / mL 6.25 μg / mL 6.36 μg / mL 6.32 μg / mL

[0113] Examples 3-4

[0114] Add 0.2 g of sodium dodecyl sulfate to 100 mL of phosphate buffer solution with pH 6.8, stir to dissolve, and prepare 3 aliquots for later use. Add 50 mg of crystal form III, crystal form I, and crystal form II of compound I to the 3 prepared aliquots, respectively. Shake for 24 hours using an air bath constant temperature shaker at 37℃ and 150 rpm. Measure the maximum apparent solubility at different time points. The results are as follows:

[0115] Crystal form 0.5h 1h 2h 6h 24h Crystal form III 113.97 μg / mL 142.67 μg / mL 121.71 μg / mL 83.46 μg / mL 66.42 μg / mL Crystal form I 61.94 μg / mL 77.15 μg / mL 79.66 μg / mL 72.58 μg / mL 56.7 μg / mL Crystal form II 48.61 μg / mL 54.16 μg / mL 55.45 μg / mL 56.83 μg / mL 52.48 μg / mL

[0116] The experimental results above show that, under the same conditions, in hydrochloric acid solution (pH 1.2) or hydrochloric acid solution containing 0.2% sodium dodecyl sulfate (pH 1.2), and in phosphate buffer solution (pH 6.8) or phosphate buffer solution containing 0.2% sodium dodecyl sulfate (pH 6.8), the dissolution rate of crystal form III of compound I is significantly faster than that of crystal form I and crystal form II of compound I. In phosphate buffer solution at pH 6.8, compared to acidic conditions, the dissolution rates of crystal forms I, II, and III of compound I are essentially the same and are not affected by pH.

[0117] Compound I has low solubility, and its dissolution is the rate-limiting step in its absorption in vivo. Therefore, increasing its dissolution rate is more conducive to its absorption in vivo, thereby improving its bioavailability and efficacy.

[0118] The foregoing has provided a detailed description of one embodiment of the present invention, but this description is merely a preferred embodiment and should not be construed as limiting the scope of the invention. All equivalent variations and modifications made within the scope of the claims of this invention should still fall within the patent coverage of this invention.

[0119] Comparative Examples

[0120] Comparative Example 1

[0121] Compound I crystals were prepared according to the methods of Examples 3 and 4 of patent CN201310183080.4. X-ray powder diffraction analysis was performed on the obtained crystals. The X-ray powder diffraction patterns all showed characteristic peaks at diffraction angles 2θ = 4.06°, 8.10°, 12.16°, 13.96°, 17.14°, 19.40°, 19.76°, 20.32°, 23.80°, 24.46°, 28.62°, and 32.26°, indicating crystal form I of compound I, as detailed in the appendix to the specification. Figure 2 As shown.

[0122] Comparative Example 2

[0123] Compound I crystals were prepared according to the methods of Examples 2, 3, 4, and 5 of patent CN201310182806.2. X-ray powder diffraction analysis was performed on the obtained crystals. The X-ray powder diffraction patterns all showed characteristic peaks at diffraction angles 2θ = 6.94°, 9.66°, 13.46°, 13.92°, 15.16°, 15.50°, 19.42°, 20.62°, 21.42°, 22.72°, 23.20°, 24.66°, 25.70°, 27.32°, and 27.78°, indicating crystal form II of compound I, as detailed in the appendix to the specification. Figure 3 As shown.

[0124] Comparative Example 3

[0125] References: "Liu Wei; Yuan Jing; Zhang Shijun et al. Synthesis, crystal structure and anticoagulant activity of" According to the description in section 2.1 of "5-chloro-N-[[(5S)-2-oxo-3-[4-(2-oxopyridin-1(2H)-yl)phenyl]oxazolidin-5-yl]methyl]thiophene-2-carboxamide[J]. Structural Chemistry. 2014, 33(7):1091─1095", compound I was prepared and crystallized in formic acid-ethanol as described. The obtained crystals were subjected to X-ray powder diffraction. The X-ray powder diffraction showed characteristic peaks at diffraction angles 2θ=4.06°, 8.10°, 12.16°, 13.96°, 17.14°, 19.40°, 19.76°, 20.32°, 23.80°, 24.46°, 28.62°, and 32.26°, which is the crystal form I of compound I.

[0126] Comparative Example 4

[0127] References: "Liu Wei; Yuan Jing; Zhang Shijun et al. Synthesis, crystal structure and anticoagulant activity of" According to the description in section 2.1 of "5-chloro-N-[[(5S)-2-oxo-3-[4-(2-oxopyridin-1(2H)-yl)phenyl]oxazolidin-5-yl]methyl]thiophene-2-carboxamide[J]. Structural Chemistry. 2014, 33(7):1091-1095", single crystals of compound I were prepared and X-ray powder diffraction was performed on the obtained crystals. The X-ray powder diffraction showed characteristic peaks at diffraction angles 2θ=6.94°, 9.66°, 13.46°, 13.92°, 15.16°, 15.50°, 19.42°, 20.62°, 21.42°, 22.72°, 23.20°, 24.66°, 25.70°, 27.32°, and 27.78°, which is the crystal form II of compound I.

[0128] Pharmacological Example 1: Prothrombin (PT) Time Model in Mice

[0129] Animals were grouped according to their body weight and tested 2 hours after oral administration. Approximately 600 μL of blood was collected from the eyes of mice and instilled into 2 ml plastic centrifuge tubes containing 60 μL of 3.8% sodium citrate. The mixture was stirred, allowed to stand for 0.5 h, and then centrifuged to separate the plasma. The proteolytic time (PT) was measured using a coagulation factor analyzer.

[0130] Effect of oral administration of crystal form III of compound I on mouse PT

[0131] Group number Group Dosage (mg / kg) route of administration PT(s) 1 blank - oral 9.41±0.51 2 Example 1-1 2.5 oral 16.78±0.91 3 Comparative Example 1 2.5 oral 16.21±1.73 4 Comparative Example 2 2.5 oral 15.80±1.49 5 Livarsaban 2.5 oral 10.35±0.64

[0132] Pharmacological Example 2: Rat model of thrombosis on arteriovenous bypass suture

[0133] Animals were grouped according to their body weight. Two hours after a single administration of urethane (1.25 g / kg, ip), the animals were anesthetized and fixed in a supine position. The right common carotid artery and left external jugular vein were isolated. A 5 cm long silk thread was inserted into the middle of a polytetrafluoroethylene (PTFE) tube. The PTFE tube was filled with heparinized saline solution (50 u / ml). After one end of the PTFE tube was inserted into the left external jugular vein, 50 u / kg of heparin was accurately injected through the PTFE tube for anticoagulation. Then, the other end of the PTFE tube was inserted into the right common carotid artery. The arterial clamp was opened, and blood flowed from the right common carotid artery into the PTFE tube and back to the left external jugular vein. Blood flow was maintained for 30 minutes, then interrupted. The silk thread was quickly removed and weighed. The wet weight of the thrombus was obtained by subtracting the weight of the silk thread from the total weight. The thrombus was dried at 60°C for 1 hour and then brought to room temperature before being weighed dry.

[0134] Effects of oral administration of crystal form III of compound I on arteriovenous bypass thrombosis

[0135] Group number Group Dosage (mg / kg) Thrombus dry weight (mg) 1 blank - 9.61±1.28 2 Example 1-1 3 6.09±1.34 3 Comparative Example 1 3 6.46±1.96 4 Comparative Example 2 3 6.82±2.47 5 Livarsaban 3 7.01±2.60 .

Claims

1. A (S)-5-chloro-N-((2-oxo-3-(4-(2-oxo-2H-pyridin-1-yl)phenyl)-1,3-oxazolidin-5-yl)methyl)thiophene-2-carboxamide (compound I) in crystal form III, characterized in that, The X-ray powder diffraction of the crystal form III exhibits characteristic peaks at diffraction angles of 2θ = 5.76 ± 0.2°, 8.30 ± 0.2°, 9.16 ± 0.2°, 11.68 ± 0.2°, 14.98 ± 0.2°, 16.38 ± 0.2°, and 16.96 ± 0.2°.

2. The (S)-5-chloro-N-((2-oxo-3-(4-(2-oxo-2H-pyridin-1-yl)phenyl)-1,3-oxazolidine-5-yl)methyl)thiophene-2-carboxamide (compound I) crystal form III as described in claim 1, characterized in that, The X-ray powder diffraction of the crystal form III exhibits characteristic peaks at diffraction angles of 2θ = 5.76 ± 0.2°, 8.30 ± 0.2°, 9.16 ± 0.2°, 11.68 ± 0.2°, 14.98 ± 0.2°, 16.38 ± 0.2°, 16.96 ± 0.2°, 21.22 ± 0.2°, 23.40 ± 0.2°, 25.20 ± 0.2°, and 26.70 ± 0.2°.

3. A method for preparing (S)-5-chloro-N-((2-oxo-3-(4-(2-oxo-2H-pyridin-1-yl)phenyl)-1,3-oxazolidine-5-yl)methyl)thiophene-2-carboxamide (compound I) crystal form III as described in claim 1 or 2, characterized in that, Includes the following steps: Compound I was dissolved in a suitable solvent to prepare a compound I drug solution. The compound I drug solution was connected to a horizontal flow pump, and the working pressure was controlled at 7.5-30 MPa. Carbon dioxide from the cylinder is introduced into the supercritical fluid antisolvent equipment system via a constant speed and constant pressure pump, and then into the crystallization kettle. The flow rate is controlled at 10-40 ml / min, the start-up temperature is controlled at 33-60℃, and the working pressure is controlled at 7.5-30 MPa. The compound I solution prepared in step (1) above is rapidly injected into the crystallization vessel through a nozzle in the supercritical fluid antisolvent equipment system using a horizontal flow pump. The flow rate is controlled at 0.05-5 ml / min. After the compound I solution is added, carbon dioxide is continuously passed through for drying. The crystals of compound I precipitated from the solvent are collected at the bottom of the crystallization vessel, and the resulting product is the crystal form III of compound I.

4. The method for preparing (S)-5-chloro-N-((2-oxo-3-(4-(2-oxo-2H-pyridin-1-yl)phenyl)-1,3-oxazolidine-5-yl)methyl)thiophene-2-carboxamide (compound I) crystal form III as described in claim 3, wherein the mass-volume ratio of compound I to solvent in step (1) is 1:15-1:50 g / ml.

5. The method for preparing (S)-5-chloro-N-((2-oxo-3-(4-(2-oxo-2H-pyridin-1-yl)phenyl)-1,3-oxazolidine-5-yl)methyl)thiophene-2-carboxamide (compound I) crystal form III as described in claim 4, wherein the mass-to-volume ratio of compound I to solvent in step (1) is 1:28-1:35 g / ml.

6. The method for preparing (S)-5-chloro-N-((2-oxo-3-(4-(2-oxo-2H-pyridin-1-yl)phenyl)-1,3-oxazolidine-5-yl)methyl)thiophene-2-carboxamide (compound I) crystal form III as described in claim 3, wherein the solvent in step (1) is selected from one or more of dimethylformamide, dimethyl sulfoxide, methanol, ethanol, acetone, dichloromethane, and tetrahydrofuran.

7. The method for preparing (S)-5-chloro-N-((2-oxo-3-(4-(2-oxo-2H-pyridin-1-yl)phenyl)-1,3-oxazolidine-5-yl)methyl)thiophene-2-carboxamide (compound I) crystal form III as described in claim 6, wherein the solvent in step (1) is selected from dimethylformamide or dimethyl sulfoxide.

8. The use of (S)-5-chloro-N-((2-oxo-3-(4-(2-oxo-2H-pyridin-1-yl)phenyl)-1,3-oxazolidin-5-yl)methyl)thiophene-2-carboxamide (compound I) crystal form III as described in any one of claims 1-2, or crystal form III prepared by the preparation method as described in any one of claims 3-7, in the preparation of drugs for the prevention / treatment of thromboembolic diseases and / or thromboembolic complications.

9. The use of (S)-5-chloro-N-((2-oxo-3-(4-(2-oxo-2H-pyridin-1-yl)phenyl)-1,3-oxazolidin-5-yl)methyl)thiophene-2-carboxamide (compound I) crystal form III as described in any one of claims 1-2, or crystal form III prepared by the preparation method as described in any one of claims 3-7, in the preparation of oral medications for the prevention / treatment of thromboembolic diseases and / or thromboembolic complications.

Images