Crystal form of compound containing alpha and beta unsaturated ketones and preparation method thereof

Abstract

The invention relates to a crystal form of a compound containing alpha and beta unsaturated ketone and a preparation method of the crystal form, and belongs to the technical field of medicine crystal forms, the compound containing alpha and beta unsaturated ketone is 5-[2, 3-dichloro-4-(2-methylene butyryl) phenoxy methylene]-3-p-trifluoromethylphenyl-1, 2, 4-oxadiazole, the crystal form comprises a crystal form A, a crystal form B and a crystal form C, the compound has good chemical stability, is easy to dissolve and high in yield and purity, and provides a basis for dosage form research and clinical application of subsequent drugs.

Description

Technical Field

[0001] The invention relates to a crystal form of a compound containing alpha, beta unsaturated ketone and a preparation method thereof, belonging to the technical field of drug crystal forms. Background Art

[0002] The anti-tumor drug candidate contains an α,β-unsaturated ketone 1,2,4-oxadiazole compound 5-[2,3-dichloro-4-(2-methylenebutyryl)phenoxymethylene]-3-p-trifluoromethylphenyl-1,2,4-oxadiazole (6u). It is a promising compound screened out from more than 100 designed and synthesized compounds. After in vivo anti-tumor activity studies and preliminary pharmacokinetic studies, it has good drugability.

[0003] The document Novel Oxadiazole Analogues Derived from Ethacrynic Acid: Design, Synthesis, and Structure-Activity Relationships in Inhibiting the Activity of Glutathione S-Transferase Pl-land Cancer Cell Proliferation. (J. Med. Chem. 2010, 53, 1015-1022.) and patent documents CN101108832A and CN106916116A disclose the preparation process of 1,2,4-oxadiazole compound 5-[2,3-dichloro-4-(2-methylenebutyryl)phenoxymethylene]-3-p-trifluoromethylphenyl-1,2,4-oxadiazole (6u).

[0004] Drug crystal form refers to the arrangement and structural form of drug molecules in the solid state. Different crystal forms of the same drug may have significant differences in appearance, solubility, melting point, dissolution, bioavailability, etc., thus affecting the stability, bioavailability and efficacy of the drug. In order to meet the needs of subsequent drug development and clinical research, it is necessary to study the crystal form of 1,2,4-oxadiazole compound 5-[2,3-dichloro-4-(2-methylenebutyryl)phenoxymethylene]-3-(p-trifluoromethylphenyl)-1,2,4-oxadiazole (6u). Summary of the invention

[0005] In view of the deficiencies of the prior art, the present invention provides a stable and soluble crystalline form of a compound containing an α, β-unsaturated ketone, wherein the compound containing an α, β-unsaturated ketone is 5-[2,3-dichloro-4-(2-methylenebutyryl)phenoxymethylene]-3-p-trifluoromethylphenyl-1,2,4-oxadiazole.

[0006] The second object of the present invention is to provide a method for preparing a compound containing an α, β-unsaturated ketone.

[0007] The present invention is achieved through the following technical solutions:

[0008] The crystalline form of the compound described in formula I is selected from one of the following: crystalline form A, crystalline form B, and crystalline form C.

[0009]

[0010] According to the preferred embodiment of the present invention, Form A, the powder X-ray diffraction pattern of Form A has a characteristic diffraction angle 2θ, interplanar spacing d and relative intensity %, and the 2θ error is ±0.2 degrees. The X-ray diffraction pattern of Form A expressed in 2θ angles has diffraction peaks at 9.66°, 14.46°, 17.84°, 19.33°, 21.55°, 22.61°, 24.17°, and 31.45°.

[0011] According to the preferred embodiment of the present invention, the TGA spectrum of Form A is as follows Figure 2 shown.

[0012] According to the preferred embodiment of the present invention, the DSC spectrum of Form A is as follows Figure 3 shown.

[0013] According to the preferred embodiment of the present invention, Form B, the powder X-ray diffraction pattern of Form B has a characteristic diffraction angle 2θ, interplanar spacing d and relative intensity %, and the 2θ error is ±0.2 degrees. The X-ray diffraction pattern of Form B expressed in 2θ angles has diffraction peaks at 9.57°, 16.61°, 18.20°, 19.15°, 21.14°, 24.11°, 24.91°, 25.80°, and 28.49°.

[0014] According to the preferred embodiment of the present invention, the TGA spectrum of Form B is as follows Figure 5 shown.

[0015] According to the preferred embodiment of the present invention, the DSC spectrum of Form B is as follows Figure 6 shown.

[0016] According to the preferred embodiment of the present invention, Form C, the powder X-ray diffraction pattern of Form C has a characteristic diffraction angle 2θ, interplanar spacing d and relative intensity %, and the 2θ error is ±0.2 degrees. The X-ray diffraction pattern of Form B expressed in 2θ angles has diffraction peaks at 9.62°, 15.60°, 16.45°, 17.94°, 18.20°, 19.38°, 21.13°, 23.96°, 24.33°, and 26.01°.

[0017] According to the preferred embodiment of the present invention, the TGA spectrum of Form C is as follows Figure 8 shown.

[0018] According to the preferred embodiment of the present invention, the DSC spectrum of Form C is as follows Fig. 9 shown.

[0019] The present invention also provides a method for preparing the crystalline form of the compound of formula I.

[0020] A method for preparing a crystalline form of the compound of formula I,

[0021] Method 1: Dissolve 5-[2,3-dichloro-4-(2-methylenebutyryl)phenoxymethylene]-3-(p-trifluoromethylphenyl)-1,2,4-oxadiazole 6u in ethanol as a single solvent, and then cool and crystallize to obtain Form A;

[0022] Method 2: 5-[2,3-dichloro-4-(2-methylenebutyryl)phenoxymethylene]-3-(p-trifluoromethylphenyl)-1,2,4-oxadiazole 6u was dissolved in acetone and evaporated naturally at room temperature to obtain Form B;

[0023] Method 3: 5-[2,3-dichloro-4-(2-methylenebutyryl)phenoxymethylene]-3-(p-trifluoromethylphenyl)-1,2,4-oxadiazole 6u was dissolved in a mixed solvent of dichloromethane and petroleum ether, and then the temperature was lowered for crystallization to obtain Form C.

[0024] Preferably according to the present invention, in method 1, the mass volume ratio of 5-[2,3-dichloro-4-(2-methylenebutyryl)phenoxymethylene]-3-p-trifluoromethylphenyl-1,2,4-oxadiazole 6u to ethanol is 1:(140-160), unit, g / mL.

[0025] Preferably, according to the present invention, in method 1, the dissolution temperature is 60°C, the cooling crystallization is cooling crystallization in a 6°C environment, and the cooling time is 6-18 hours, preferably 12 hours.

[0026] Preferably according to the present invention, in method 2, the amount of acetone added is 4 to 40 mL of acetone per 500 mg of 5-[2,3-dichloro-4-(2-methylenebutyryl)phenoxymethylene]-3-p-trifluoromethylphenyl-1,2,4-oxadiazole 6u.

[0027] Preferably according to the present invention, in method 3, the volume ratio of the mixed solvent of dichloromethane and petroleum ether is (1-5):(40-60).

[0028] Preferably according to the present invention, in method 3, the mass volume ratio of 5-[2,3-dichloro-4-(2-methylenebutyryl)phenoxymethylene]-3-(p-trifluoromethylphenyl)-1,2,4-oxadiazole 6u to a mixed solvent of dichloromethane and petroleum ether is 1:(100-120), unit, g / mL.

[0029] Preferably, according to the present invention, in method 3, the cooling crystallization is cooling crystallization in a 6°C environment, and the cooling time is 1-3 hours, preferably 3 hours.

[0030] The present invention has the following beneficial effects:

[0031] 1. The crystalline forms A, B and C of the compound of the present invention all have good chemical stability, high yield and purity, and provide a basis for subsequent drug dosage form research and clinical application.

[0032] 2. The preparation method of the crystal form of the present invention is simple and easy, can be stably and mass-produced, and is conducive to popularization and application.

[0033] 3. The crystal form of the present invention has the advantages of high solubility in water and good stability. BRIEF DESCRIPTION OF THE DRAWINGS

[0034] Figure 1 is the powder X-ray diffraction pattern of Form A obtained in Example 1;

[0035] Figure 2 This is the thermogravimetric analysis spectrum TGA of Form A obtained in Example 1;

[0036] Figure 3 The differential scanning calorimetry (DSC) spectrum of Form A obtained in Example 1;

[0037] Figure 4 is the powder X-ray diffraction pattern of Form B obtained in Example 2;

[0038] Figure 5 This is the thermogravimetric analysis spectrum TGA of Form B obtained in Example 2;

[0039] Figure 6 The differential scanning calorimetry (DSC) spectrum of Form B obtained in Example 2;

[0040] Figure 7 The X-ray powder diffraction pattern of the crystal form C obtained in Example 3;

[0041] Figure 8 This is the thermogravimetric analysis spectrum TGA of Form C obtained in Example 3;

[0042] Fig. 9 This is the differential scanning calorimetry (DSC) spectrum of Form C obtained in Example 3. DETAILED DESCRIPTION

[0043] The present invention will be further described below with reference to the accompanying drawings and in combination with specific embodiments. The embodiments are provided to better illustrate the content of the present invention, but the content of the present invention is not limited to the embodiments. Therefore, those skilled in the art may make non-essential improvements and adjustments to the implementation scheme according to the above invention content, which still fall within the protection scope of the present invention.

[0044] In the embodiment:

[0045] Preparation method of 5-[2,3-dichloro-4-(2-methylenebutyryl)phenoxymethylene]-3-p-trifluoromethylphenyl-1,2,4-oxadiazole 6u:

[0046] 1) 2,3-Dichloroanisole (21.2g, 120mmol) was placed in a reaction bottle, and 120mL of dichloromethane was added to dissolve it. The mixture was cooled to 0℃ in an ice bath, and aluminum chloride (16.8g, 126mmol) was added. The mixture was stirred magnetically for 0.5h. Butyryl chloride (13.4g, 126mmol) was slowly added with a constant pressure dropping funnel. The mixture was reacted for 0.5h in an ice bath, and then heated to room temperature for 2.5h. The solvent was evaporated under reduced pressure, and dichloromethane was added. The solvent was evaporated under reduced pressure twice. Then 120mL of dichloromethane and aluminum chloride (13.4g, 126mmol) were added in sequence, and the mixture was heated to 40℃ for 3h. The temperature was lowered to room temperature, the reaction flask was placed in an ice bath, 240 mL of ice water was slowly added dropwise to the reaction flask, the organic phase was separated, the ice-water phase was extracted with dichloromethane three times, the organic phases were combined, washed twice with saturated sodium chloride, dried over anhydrous sodium sulfate (still anhydrous sodium sulfate), filtered, and the solvent was evaporated under reduced pressure to obtain 2,3-dichloro-4-butyrylphenol.

[0047] 2) 2,3-dichloro-4-butyrylphenol (26.0 g, 111.5 mmol) was placed in a reaction flask, 120 mL of acetone was added to dissolve it, potassium carbonate (16.9 g, 122.7 mmol) and potassium iodide (3.7 g, 22.3 mmol) were added in sequence, magnetic stirring was performed for 0.5 h, ethyl bromoacetate (20.5 g, 122.7 mmol) was added dropwise, the temperature was raised to 60 °C for reaction for 4 h, the temperature was lowered to room temperature, acetone was evaporated, water and ethyl acetate were added, the aqueous phase was separated, the organic phase was washed with water twice, the solvent was evaporated under reduced pressure to obtain an oily substance, 80 mL of ethanol was added to dissolve it, 80 mL of water and potassium hydroxide (12.5 g, 223.0 mmol) were added in sequence, the reaction was performed at room temperature for 2 h, ethanol was evaporated under reduced pressure, concentrated hydrochloric acid was added dropwise to pH = 1, a large amount of white solid was precipitated, filtered, and dried to obtain 2-[2,3-dichloro-4-butyrylphenoxy]acetic acid.

[0048] 3) 2-[2,3-dichloro-4-butyrylphenoxy]acetic acid (30.0 g, 103.1 mmol) was placed in a reaction flask, 100 mL of ethanol and 100 mL of water were added to dissolve it, potassium carbonate (28.5 g, 206.2 mmol) and paraformaldehyde (6.2 g, 206.2 mmol) were added in sequence, and the mixture was reacted at 90°C for 10 h, cooled to room temperature, ethanol was evaporated under reduced pressure, concentrated hydrochloric acid was added dropwise until pH = 1, a white precipitate was precipitated, filtered, dried, and recrystallized from petroleum ether / ethyl acetate to obtain 2-[2,3-dichloro-4-(2-methylenebutyryl)phenoxy]acetic acid as a white powder.

[0049] 4) Preparation of trifluoromethylbenzamide oxime:

[0050] Place p-trifluoromethylbenzonitrile (3.4 g, 20.0 mmol) in a reaction bottle, add 30 mL of methanol, add hydroxylamine hydrochloride (2.8 g, 40.0 mmol) and triethylamine (4.0 g, 40.0 mmol) in sequence, react at room temperature for 0.5 h, evaporate the solvent under reduced pressure, dissolve the residue with water and ethyl acetate, extract the aqueous phase twice with ethyl acetate, combine the organic phases, wash the organic phases twice with saturated sodium chloride, dry over anhydrous sodium sulfate, filter, and evaporate ethyl acetate under reduced pressure to obtain p-trifluoromethylbenzamide oxime.

[0051] 5) Preparation of 6u:

[0052] Place 2-[2,3-dichloro-4-(2-methylenebutyryl)phenoxy]acetic acid (1.0 g, 3.3 mmol) in a reaction flask, add 10 mL of dichloromethane, dropwise add oxalyl chloride (0.84 g, 6.6 mmol), and dropwise add two drops of N,N-dimethylformamide. After 0.5 h at room temperature, evaporate the solvent under reduced pressure to obtain a light yellow oil, add 10 mL of acetone to dissolve it for later use;

[0053] Place p-trifluoromethylbenzamide oxime (0.74 g, 3.6 mmol) in a reaction flask, add 10 mL of acetone to dissolve, add potassium carbonate (0.91 g, 6.6 mmol), stir at room temperature for 2 h, slowly drop the above-mentioned acyl chloride in acetone solution under ice bath conditions, react at room temperature for 3 h, heat to 60 ° C to react for 5 h, cool to room temperature, evaporate acetone, dissolve the residue in water and ethyl acetate, separate the aqueous phase, extract the aqueous phase twice with ethyl acetate, combine the organic phases, wash the organic phases twice with water and saturated sodium chloride solution, dry over anhydrous sodium sulfate, filter, and evaporate the solvent under reduced pressure to obtain a crude product of 5-[2,3-dichloro-4-(2-methylenebutyryl)phenoxymethylene]-3-p-trifluoromethylphenyl-1,2,4-oxadiazole, which was purified by column chromatography with petroleum ether / ethyl acetate (8:1-7:1) to obtain 6u.

[0054] Example 1

[0055] Preparation method of crystal form A:

[0056] Take 500 mg of 5-[2,3-dichloro-4-(2-methylenebutyryl)phenoxymethylene]-3-(p-trifluoromethylphenyl)-1,2,4-oxadiazole 6u, put it into a 250 mL eggplant-shaped bottle, add 78 mL of ethanol, heat in a 60°C water bath until the solution is clear, cool to 6°C and keep warm for 12 h. After crystallization is completed, filter, wash, and dry at room temperature to obtain a white solid with a yield of 89% and a purity of 98.4%.

[0057] The crystalline form A prepared in Example 1 has the following characteristics: Figure 1 The X-ray powder diffraction pattern is shown.

[0058] The thermogravimetric analysis (TGA) spectrum of the obtained crystal form A is as follows: Figure 2 As shown in the figure, it can be seen that Form A starts to lose weight when heated to 117°C; when heated to 580°C, the weight loss is about 93.76%.

[0059] The differential scanning calorimetry (DSC) of the obtained crystal form A is as follows: Figure 3 As shown, there is no endothermic peak at 0-280°C, indicating that Form A is stable at this temperature.

[0060] Example 2

[0061] Preparation method of crystal form B:

[0062] Take 5-[2,3-dichloro-4-(2-methylenebutyryl)phenoxymethylene]-3-(p-trifluoromethylphenyl)-1,2,4-oxadiazole 6u 500mg, put it into a 100mL eggplant-shaped bottle, add 5mL of acetone, all the solids are dissolved, leave it at room temperature without cover for 3h, a large amount of solids precipitate, crystallization is completed, filter, wash, and dry at room temperature to obtain a white solid with a yield of 93% and a purity of 97.4%.

[0063] The crystalline form B prepared in Example 2 has the following characteristics: Figure 4 The X-ray powder diffraction pattern is shown.

[0064] The thermogravimetric analysis (TGA) spectrum of the obtained crystal form B is as follows: Figure 5 As shown in the figure, it can be seen that Form B starts to lose weight when heated to 95°C; when heated to 580°C, the weight loss is about 90.02%.

[0065] The differential scanning calorimetry (DSC) of the obtained crystal form B is as follows: Figure 6 As shown, there is no endothermic peak at 0-280°C, indicating that Form B is stable at this temperature.

[0066] Example 3

[0067] Preparation method of crystal form C:

[0068] Take 5-[2,3-dichloro-4-(2-methylenebutyryl)phenoxymethylene]-3-(p-trifluoromethyl)phenyl-1,2,4-oxadiazole 6u 500mg, put it into a 250mL eggplant-shaped bottle, add 15mL of dichloromethane, all the solids are dissolved, slowly add 40mL of petroleum ether along the wall of the bottle, the solution is clear, cool to 6°C, keep warm for 3h, white solid precipitates, crystallization is completed, filter, wash, and dry at room temperature to obtain a white solid with a yield of 95% and a purity of 98.2%.

[0069] The crystalline form C prepared in Example 3 has the following characteristics: Figure 7 The X-ray powder diffraction pattern is shown.

[0070] The thermogravimetric analysis (TGA) spectrum of the obtained crystal form C is as follows: Figure 8 As shown in the figure, it can be seen that Form C starts to lose weight when heated to 184°C; when heated to 580°C, the weight loss is about 95.81%.

[0071] The differential scanning calorimetry (DSC) of the obtained crystal form C is as follows: Fig. 9 As shown, there is an endothermic peak at 120°C, indicating that Form C melts at 120°C.

[0072] Experimental example

[0073] Solubility test method: Grind the crystal forms A, B and C into fine powders respectively, take 0.1g of each and add them to the solvents described in Table 1, shake vigorously for 30s every 5min and observe the dissolution within 30 minutes. If there is solid, continue to add liquid until there is no precipitation. The test results are shown in Table 1.

[0074] Very soluble means that 1g (ml) of solute can be dissolved in less than 1ml of solvent;

[0075] Soluble means that 1g (ml) of solute can be dissolved in 1 to less than 10ml of solvent;

[0076] Solubility means that 1g (ml) of solute can be dissolved in 10 to less than 30ml of solvent;

[0077] Slightly soluble means that 1g (ml) of the solute can be dissolved in 30 to less than 100ml of the solvent;

[0078] Slightly soluble means that 1g (ml) of solute can be dissolved in 100 to less than 1000ml of solvent;

[0079] Very slightly soluble means that 1g (ml) of solute can be dissolved in 1000 to less than 10000ml of solvent;

[0080] Almost insoluble or insoluble means that 1g (ml) of the solute cannot be completely dissolved in 10000ml of the solvent.

[0081] Table 1

[0082] Solvents Crystalline Form A Form B Crystalline Form C Ethyl acetate Soluble Soluble Soluble Methanol Insoluble Insoluble Insoluble Dichloromethane Soluble Soluble Soluble 0.1mol / L hydrochloric acid Insoluble Insoluble Insoluble 0.1mol / L sodium hydroxide Insoluble Insoluble Insoluble

[0083] As can be seen from Table 1, Forms A, B and C are easily soluble in ethyl acetate and dichloromethane, but insoluble in other solvents.

[0084] Purity determination: (HPLC method)

[0085] Instrument model: Shimadzu;

[0086] Column: Eclipse Plus-C18, 5 μM, 4.6 × 250 mm;

[0087] Mobile phase: methanol: water = 10:1;

[0088] Flow rate 1ml / min;

[0089] Sample: the samples prepared in Examples 1, 2, and 3;

[0090] Experimental method: Take 2 mg of the samples obtained in Examples 1, 2, and 3 above, respectively, add 3 mL of chromatographic methanol to dissolve; filter with a microporous filter membrane, and inject the sample into the injector;

[0091] Experimental results: The purity of Form A in Example 1 is 98.4%; the purity of Form B in Example 2 is 97.4%; the purity of Form C in Example 3 is 98.2%.

Claims

1. A crystalline form of the compound of formula I, wherein the crystalline form is selected from one of the following: crystalline form A, crystalline form B, and crystalline form C; Form A, the powder X-ray diffraction pattern of Form A has characteristic diffraction angle 2θ, interplanar spacing d and relative intensity %, and the 2θ error is ±0.2 degrees. The X-ray diffraction pattern of Form A expressed in 2θ angle has diffraction peaks at 9.66°, 14.46°, 17.84°, 19.33°, 21.55°, 22.61°, 24.17°, and 31.45°; Form B, the powder X-ray diffraction pattern of Form B has characteristic diffraction angle 2θ, interplanar spacing d and relative intensity %, and the 2θ error is ±0.2 degrees. The X-ray diffraction pattern of Form B expressed in 2θ angle has diffraction peaks at 9.57°, 16.61°, 18.20°, 19.15°, 21.14°, 24.11°, 24.91°, 25.80°, and 28.49°; According to the preferred embodiment of the present invention, Form C, the powder X-ray diffraction pattern of Form C has a characteristic diffraction angle 2θ, interplanar spacing d and relative intensity %, and the 2θ error is ±0.2 degrees. The X-ray diffraction pattern of Form B expressed in 2θ angles has diffraction peaks at 9.62°, 15.60°, 16.45°, 17.94°, 18.20°, 19.38°, 21.13°, 23.96°, 24.33°, and 26.01°.

2. A method for preparing the crystalline form of the compound of formula I according to claim 1, Method 1: Dissolve 5-[2,3-dichloro-4-(2-methylenebutyryl)phenoxymethylene]-3-(p-trifluoromethylphenyl)-1,2,4-oxadiazole 6u in ethanol as a single solvent, and then cool and crystallize to obtain Form A; Method 2: 5-[2,3-dichloro-4-(2-methylenebutyryl)phenoxymethylene]-3-(p-trifluoromethylphenyl)-1,2,4-oxadiazole 6u was dissolved in acetone and evaporated naturally at room temperature to obtain Form B; Method 3: 5-[2,3-dichloro-4-(2-methylenebutyryl)phenoxymethylene]-3-(p-trifluoromethylphenyl)-1,2,4-oxadiazole 6u was dissolved in a mixed solvent of dichloromethane and petroleum ether, and then the temperature was lowered for crystallization to obtain Form C.

3. The preparation method according to claim 2, characterized in that: In method 1, the mass volume ratio of 5-[2,3-dichloro-4-(2-methylenebutyryl)phenoxymethylene]-3-(p-trifluoromethylphenyl)-1,2,4-oxadiazole 6u to ethanol is 1:(140-160), unit, g / mL.

4. The preparation method according to claim 2, characterized in that: In method 1, the dissolving temperature is 60°C, the cooling crystallization is cooling crystallization in a 6°C environment, and the cooling time is 6-18 hours, preferably 12 hours.

5. The preparation method according to claim 2, characterized in that: In method 2, the amount of acetone added is 4 to 40 mL for every 500 mg of 5-[2,3-dichloro-4-(2-methylenebutyryl)phenoxymethylene]-3-(p-trifluoromethylphenyl)-1,2,4-oxadiazole 6u.

6. The preparation method according to claim 2, characterized in that: In method 3, the mixed solvent of dichloromethane and petroleum ether has a volume ratio of dichloromethane to petroleum ether of (1-5):(40-60).

7. The preparation method according to claim 2, characterized in that: In method 3, the mass volume ratio of 5-[2,3-dichloro-4-(2-methylenebutyryl)phenoxymethylene]-3-(p-trifluoromethylphenyl)-1,2,4-oxadiazole 6u to a mixed solvent of dichloromethane and petroleum ether is 1:(100-120), unit, g / mL.

8. The preparation method according to claim 2, characterized in that: In method 3, the cooling crystallization is carried out in a 6°C environment, and the cooling time is 1-3 hours.

9. The preparation method according to claim 2, characterized in that: In method 3, the cooling time is 3 hours.

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