CRYSTAL FORM OF AROMATIC VINYL DERIVATIVES, AND PREPARATION METHOD THEREFOR AND USE THEREOF

Abstract

Disclosed in the present invention are a crystal form of aromatic vinyl derivatives, and a preparation method therefor and the use thereof. Specifically disclosed in the present invention are crystal form A, crystal form B and crystal form C of a compound as shown in formula I. The crystal forms of the present invention have a good stability, are less hygroscopic and are easy to prepare, and have an important value in terms of the optimization and development of drugs.

##STR00001##

Claims

1. A crystal form A of a compound as shown in formula I, the X-ray powder diffraction pattern of the crystal form A represented by 2θ angles has characteristic peaks at: 9.923±0.2°, 10.883±0.2° and 17.357±0.2°; or, has characteristic peaks at 3.979±0.2°, 9.923±0.2°, 10.883±0.2°, 17.357±0.2°, 18.607±0.2° and 19.294±0.2°; or, has characteristic peaks at 3.979±0.2°, 4.991±0.2°, 9.923±0.2°, 10.883±0.2°, 14.251±0.2°, 16.210±0.2°, 17.357±0.2°, 18.607±0.2°, 19.294±0.2°, 19.594±0.2° and 20.792±0.2°; or, has characteristic peaks at 3.979±0.2°, 4.991±0.2°, 7.113±0.2°, 8.135±0.2°, 9.923±0.2°, 10.883±0.2°, 11.613±0.2°, 14.251±0.2°, 14.866±0.2°, 16.210±0.2°, 17.357±0.2°, 18.607±0.2°, 19.294±0.2°, 19.594±0.2°, 20.792±0.2°, 21.272±0.2°, 24.437±0.2°, 25.257±0.2°, 26.2295±0.2°, 27.870±0.2°, 28.631±0.2°, 29.126±0.2°, 29.943±0.2°; ##STR00006##

2. The crystal form A of the compound as shown in formula I as defined in claim 1, wherein, in the X-ray powder diffraction pattern of the crystal form A represented by 2θ angles, the 2θ values are as shown in the following table; TABLE-US-00018 2θ Relative (2θ ± 0.2°) intensity (%) 3.979 25.2 4.991 14.3 7.113 1.5 8.135 1.5 9.923 36.3 10.883 27.0 11.613 6.6 14.251 13.2 14.866 4.4 16.210 18.0 17.357 100.0 18.607 25.0 19.294 26.3 19.594 11.4 20.792 11.2 21.272 6.8 24.437 6.7 25.257 5.9 26.229 4.4 27.870 3.7 28.631 1.9 29.126 5.2 29.943 1.5 or, in the polarized light microscope analysis of the crystal form A, the shape of the crystal form is granular or rod-shaped; or, in the differential scanning calorimetry analysis of the crystal form A, the differential scanning calorimetry analysis of the crystal form A has a thermal absorption peak at 247° C.; or, in the thermogravimetric analysis of the crystal form A, the sample has a weight loss of only 0.1447% from 26.76° C. to 119.97° C., and the “%” is the weight percentage; or, in the dynamic vapor sorption analysis of the crystal form A, the hygroscopic weight gain is 0.310% at 80% RH and the hygroscopic weight gain is 0.409% at 95% RH.

3. A crystal form B of the compound as shown in formula I, the X-ray powder diffraction pattern of the crystal form B represented by 2θ angles has characteristic peaks at 3.424±0.2°, 6.576±0.2° and 19.297±0.2°; or, has characteristic peaks at 3.424±0.2°, 6.576±0.2°, 18.217±0.2°, 19.297±0.2°, 20.901±0.2° and 26.379±0.2°; or, has characteristic peaks at 3.424±0.2°, 6.576±0.2°, 14.467±0.2°, 16.406±0.2°, 17.567±0.2°, 18.217±0.2°, 19.297±0.2°, 20.557±0.2°, 20.901±0.2°, 22.460±0.2°, 25.084±0.2°, 25.878±0.2°, 26.379±0.2° and 28.983±0.2°; or, has characteristic peaks at 3.424±0.2°, 6.576±0.2°, 9.732±0.2°, 11.304±0.2°, 12.905±0.2°, 13.918±0.2°, 14.467±0.2°, 16.406±0.2°, 17.567±0.2°, 18.217±0.2°, 19.297±0.2°, 20.557±0.2°, 20.901±0.2°, 22.460±0.2°, 23.872±0.2°, 25.084±0.2°, 25.878±0.2°, 26.379±0.2°, 28.983±0.2°, 29.531±0.2°, 30.459±0.2°, 32.171±0.2°, 34.297±0.2°, 37.676±0.2° and 38.902±0.2°; ##STR00007##

4. The crystal form B of the compound as shown in formula I as defined in claim 3, wherein, in the X-ray powder diffraction pattern of the crystal form B represented by 2θ angles, the 2θ values are as shown in the following table; TABLE-US-00019 2θ Relative (2θ ± 0.2°) intensity (%) 3.424 89.1 6.576 100.0 9.732 3.4 11.304 3.7 12.905 3.2 13.918 8.7 14.467 13.9 16.406 17.6 17.567 11.6 18.217 26.6 19.297 87.8 20.557 16.2 20.901 35.7 22.460 24.7 23.872 8.3 25.084 12.1 25.878 20.0 26.379 25.5 28.983 10.2 29.531 8.0 30.459 1.6 32.171 2.3 34.297 2.0 37.676 1.9 38.902 1.6 or, in the differential scanning calorimetry analysis of the crystal form B, the differential scanning calorimetry analysis of the crystal form B has a thermal absorption peak at 243° C.; or, in the thermogravimetric analysis of the crystal form B, the sample has a weight loss of 5.2% from 25.3° C. to 92.5° C., and the “%” is the weight percentage; or, in the dynamic vapor sorption analysis of the crystal form B, the sample has a weight gain of 7.235% from 0% RH to 95% RH.

5. A crystal form C of the compound as shown in formula I, the X-ray powder diffraction pattern of the crystal form C represented by 2θ angles has characteristic peaks at 6.250±0.2°, 18.458±0.2° and 19.302±0.2°; or, has characteristic peaks at 6.250±0.2°, 8.779±0.2°, 13.720±0.2°, 18.458±0.2° and 19.302±0.2°; or, has characteristic peaks at 6.250±0.2°, 8.779±0.2°, 12.635±0.2°, 13.720±0.2°, 16.525±0.2°, 18.458±0.2° and 19.302±0.2°; or, has characteristic peaks at 6.250±0.2°, 6.979±0.2°, 8.779±0.2°, 12.635±0.2°, 13.720±0.2°, 16.525±0.2°, 18.458±0.2°, 19.302±0.2°, 20.852±0.2°, 22.345±0.2°, 24.772±0.2°, 25.230±0.2° and 27.285±0.2°; or, has characteristic peaks at 6.250±0.2°, 6.979±0.2°, 8.779±0.2°, 12.635±0.2°, 13.720±0.2°, 15.285±0.2°, 16.525±0.2°, 18.458±0.2°, 19.302±0.2°, 20.852±0.2°, 22.345±0.2°, 24.772±0.2°, 25.230±0.2°, 25.996±0.2°, 27.285±0.2°, 28.303±0.2°, 28.829±0.2°, 29.699±0.2°, 30.703±0.2°, 33.133±0.2°, 34.655±0.2°, 36.829±0.2°, 37.967±0.2°; ##STR00008##

6. The crystal form C of the compound as shown in formula I as defined in claim 5, wherein, in the X-ray powder diffraction pattern of the crystal form C represented by 2θ angles, the 2θ values are as shown in the following table; TABLE-US-00020 2θ Relative (2θ ± 0.2°) intensity (%) 6.250 100 6.979 15.3 8.779 38.6 12.635 21.3 13.720 32.6 15.285 3.1 16.525 24.1 17.696 5.5 18.458 73.0 19.302 59.0 20.852 15.4 22.345 19.1 24.772 14.3 25.230 11.5 25.996 4.1 27.285 13.2 28.303 4.1 28.829 3.7 29.699 2.3 30.703 5.6 33.133 3.1 34.655 1.9 36.829 2.4 37.967 3.1 or, in the differential scanning calorimetry analysis of the crystal form C, the differential scanning calorimetry analysis of the crystal form C has a thermal absorption peak at 243° C., and the melting heat is preferably 99.33 J/g; or, in the thermogravimetric analysis of the crystal form C, the sample has a weight loss of 0.62% from 24.0° C. to 58.0° C., and the sample has a weight loss of 2.5% from 58.0° C. to 162.3° C., and the “%” is the weight percentage; or, in the dynamic vapor sorption analysis of the crystal form C, the sample has a weight gain of 4.767% from 0% RH to 95% RH.

7. The crystal form A of the compound as shown in formula I as defined in claim 1, wherein the polarized light micrograph of the crystal form A is as shown in FIG. 1; or the differential scanning calorimetry analysis pattern of the crystal form A is as shown in FIG. 2; or, the thermogravimetric analysis pattern of the crystal form A is as shown in FIG. 3; or, the X-ray powder diffraction pattern of the crystal form A is as shown in FIG. 4; or, the dynamic vapor sorption analysis pattern of the crystal form A is as shown in FIG. 5.

8. A method of preparing the crystal form A of the compound as shown in formula I as defined in claim 1, comprising the following steps: in a solvent, crystallizing the compound as shown in formula I; the crystallization method is suspension equilibrium method, solution heating-slow cooling method or anti-solvent method; the solvent is ethanol, and when the crystallization method is anti-solvent method, the anti-solvent is an alkane solvent.

9. The method of preparing the crystal form A of the compound as shown in formula I as defined in claim 8, wherein, the crystallization temperature is 20° C. to 60 ° C., for example, room temperature or 50° C.; or, when the crystallization method is anti-solvent method, the alkane solvent is C.sub.1-10 alkane solvent, preferably n-heptane; or, the mass-volume ratio of the compound as shown in formula Ito the solvent is 5 mg/mL to 20 mg/mL, for example, 7.7 mg/mL to 20 mg/mL; or, the crystallization time is 1 hour to 20 days, for example, 1 hour to 2 hours, 5 hours to 6 hours or 10 days to 20 days; or, when the crystallization method is anti-solvent method, the mass ratio of the anti-solvent to the solvent is 5:1 to 8:1, for example, 6.5:1.

10. A method of preparing the crystal form B of the compound as shown in formula I as defined in claim 3, comprising the following steps: in a solvent, crystallizing the compound as shown in formula I; the crystallization method is suspension equilibrium method or anti-solvent method; when the crystallization method is suspension equilibrium method, the solvent is water, or ethanol and water; and when the crystallization method is anti-solvent method, the solvent is ethanol or tetrahydrofuran, and the anti-solvent is water.

11. The method of preparing the crystal form B of the compound as shown in formula I as defined in claim 10, wherein, the water is one or more of distilled water, deionized water, purified water, tap water and mineral water; or, the crystallization temperature is 20° C. to 60° C., for example, room temperature or 50° C.; or, the mass-volume ratio of the compound as shown in formula Ito the solvent is 5 mg/mL to 40 mg/mL, for example, 7.7 mg/mL, 11.1 mg/mL, 20 mg/mL or 33.3 mg/mL; or, the crystallization time is 1 hour to 20 days, for example, 1 hour to 2 hours, 1 day or 17 days; or, when the crystallization method is suspension equilibrium method, and when the solvent is ethanol and water, the volume ratio of ethanol to water is 1:3 to 1:5, for example, 1:4; or, when the crystallization method is anti-solvent method, the volume ratio of the anti-solvent to the solvent is 1:1 to 4:1, for example, 1:1 or 2.7:1.

12. A method of preparing the crystal form C of the compound as shown in formula I as defined in claim 5, comprising the following steps: in a solvent, crystallizing the compound as shown in formula I by suspension equilibrium method; the solvent is isopropanol, N,N-dimethylacetamide, or acetone and water.

13. The method of preparing the crystal form C of the compound as shown in formula I as defined in claim 12, wherein, the water is one or more of distilled water, deionized water, purified water, tap water and mineral water; or, when the solvent is acetone and water, the volume ratio of acetone to water is 7:1 to 10:1, for example, 8:1; or, the crystallization temperature is room temperature; or, the mass-volume ratio of the compound as shown in formula Ito the solvent is 10 mg/mL to 50 mg/mL, for example, 20 mg/mL, 40 mg/mL or 44.4 mg/mL; or, the crystallization time is 1 day to 20 days, for example, 1 day, 7 days, 10 days or 20 days.

14. A method for treating cancer in a subject in need thereof comprising: administering an effective amount of the crystal form A of the compound as shown in formula I as defined in claim 1 to the subject the cancer is preferably one or more of lung cancer, esophageal cancer. gastric cancer, colorectal cancer, liver cancer, nasopharyngeal cancer, brain tumor, breast cancer, cervical cancer, blood cancer and bone

15. The crystal form A of the compound as shown in formula I as defined in claim 2, wherein, in the polarized light microscope analysis of the crystal form A, the particle size of the crystal form A is 10 to 100 μm; or, in the differential scanning calorimetry analysis of the crystal form A, the melting heat is 118.0 J/g.

16. The crystal form B of the compound as shown in formula I as defined in claim 4, wherein, in the differential scanning calorimetry analysis of the crystal form B, the melting heat is 93.73 J/g.

17. The crystal form B of the compound as shown in formula I as defined in claim 3, wherein the X-ray powder diffraction pattern of the crystal form B is as shown in FIG. 6; or, the differential scanning calorimetry analysis pattern of the crystal form B is as shown in FIG. 7; or, the thermogravimetric analysis pattern of the crystal form B is as shown in FIG. 8; or, the dynamic vapor sorption analysis pattern of the crystal form B is as shown in FIG. 9.

18. The crystal form C of the compound as shown in formula I as defined in claim 5, wherein the X-ray powder diffraction pattern of the crystal form C is as shown in FIG. 10; or, the differential scanning calorimetry analysis pattern of the crystal form C is as shown in FIG. 11; or, the thermogravimetric analysis pattern of the crystal form C is as shown in FIG. 12; or, the dynamic vapor sorption analysis pattern of the crystal form C is as shown in FIG. 13.

19. A method for treating cancer in a subject in need thereof, comprising: administering an effective amount of the crystal form B of the compound as shown in formula I as defined in claim 3 to the subject; the cancer is preferably one or more of lung cancer, esophageal cancer, gastric cancer, colorectal cancer, liver cancer, nasopharyngeal cancer, brain tumor, breast cancer, cervical cancer, blood cancer and bone cancer.

20. A method for treating cancer in a subject in need thereof, comprising: administering an effective amount of the crystal form C of the compound as shown in formula I as defined in claim 5 to the subject; the cancer is preferably one or more of lung cancer, esophageal cancer, gastric cancer, colorectal cancer, liver cancer, nasopharyngeal cancer, brain tumor, breast cancer, cervical cancer, blood cancer and bone cancer.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

[0074] FIG. 1 is the polarized light micrograph of the crystal form A of the compound as shown in formula I.

[0075] FIG. 2 is the DSC pattern of the crystal form A of the compound as shown in formula I.

[0076] FIG. 3 is the TGA pattern of the crystal form A of the compound as shown in formula I.

[0077] FIG. 4 is the XRPD pattern of the crystal form A of the compound as shown in formula I.

[0078] FIG. 5 is the DVS pattern of the crystal form A of the compound as shown in formula I.

[0079] FIG. 6 is the XRPD pattern of the crystal form B of the compound as shown in formula I.

[0080] FIG. 7 is the DSC pattern of the crystal form B of the compound as shown in formula I.

[0081] FIG. 8 is the TGA pattern of the crystal form B of the compound as shown in formula I.

[0082] FIG. 9 is the DVS pattern of the crystal form B of the compound as shown in formula I.

[0083] FIG. 10 is the XRPD pattern of the crystal form C of the compound as shown in formula I.

[0084] FIG. 11 is the DSC pattern of the crystal form C of the compound as shown in formula I.

[0085] FIG. 12 is the TGA pattern of the crystal form C of the compound as shown in formula I.

[0086] FIG. 13 is the DVS pattern of the crystal form C of the compound as shown in formula I.

[0087] FIG. 14 is the XRPD pattern of the crystal form V of the compound as shown in formula I.

[0088] FIG. 15 is the DSC pattern of the crystal form V of the compound as shown in formula I.

[0089] FIG. 16 is the TGA pattern of the crystal form V of the compound as shown in formula I.

[0090] FIG. 17 is the XRPD pattern of the crystal form VII of the compound as shown in formula I.

[0091] FIG. 18 is the DSC pattern of the crystal form VII of the compound as shown in formula I.

[0092] FIG. 19 is the TGA pattern of the crystal form VII of the compound as shown in formula I.

[0093] FIG. 20 is the XRPD pattern of the crystal form VIII of the compound as shown in formula I.

[0094] FIG. 21 is the DSC pattern of the crystal form VIII of the compound as shown in formula I.

[0095] FIG. 22 is the TGA pattern of the crystal form VIII of the compound as shown in formula I.

[0096] FIG. 23 is the XRPD pattern of the crystal form IX of the compound as shown in formula I.

[0097] FIG. 24 is the DSC pattern of the crystal form IX of the compound as shown in formula I.

[0098] FIG. 25 is the TGA pattern of the crystal form IX of the compound as shown in formula I.

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENT

[0099] The present disclosure is further described below by way of embodiments, but the present disclosure is not thereby limited to the scope of the described embodiments. The experimental methods not specified in the specific conditions in the following embodiments are selected according to the conventional methods and conditions, or according to the commodity instructions.

[0100] In the preparation method of the following crystal forms, the volume (mL) of the added solvent=sample mass×volume multiple (mL), for example, the volume of ethanol in the preparation method I of embodiment 1 is: 0.02×50 mL=1 mL.

[0101] Preparation of amorphous compound I (the compound as shown in formula I):

[0102] At room temperature, glacial acetic acid (32.8 mg, 0.54 mmol) was added to a mixed solution of compound 5-a (preparation of compound 5-a was obtained according to the method in patent CN109988144A) and (S)-2-methylserine (65 mg, 0.54 mmol) in methanol (10 mL) and dichloromethane (10 mL), and the reaction solution was stirred at room temperature for 1 hour. Then, sodium cyanoborohydride (85.8 mg, 1.36 mmol) was added thereto and the mixture was stirred for 16 hours. The reaction solution was concentrated under reduced pressure, and the residue was dissolved in ethyl acetate (50 mL), washed with water (20 mL) and saturated brine (20 mL), and then the organic phase was dried over anhydrous sodium sulfate, concentrated under reduced pressure. The residue was separated by silica gel thin layer chromatography preparative plate with the mobile phase of dichloromethane: methanol=10:1, and the silica gel mixed with compound I was washed with methanol (20 mL×3), and the washing solution was concentrated under reduced pressure to obtain amorphous compound I (24 mg, yield: 18.7%). LC-MS (ESI): m/z=468 [M—H].sup.−

[0103] .sup.1H NMR (500 MHz, CD.sub.3OD)δ: 8.17 (s, 1H), 7.81-7.79 (d, J=8.5 Hz, 1H), 7.68-7.62 (m, 2H), 7.57-7.55 (d, J=8.0 Hz, 1H), 7.46-7.43 (m, 2H), 7.39-7.28 (m, 5H), 7.20-7.19 (d, J=7.0 Hz, 1H), 4.36-4.28 (q,2H), 4.03-4.00 (d, J=12.5 Hz, 1H), 3.86-3.84 (d, J=12.5 Hz, 1H), 2.33 (s,3H), 1.57 (s,3H) ppm.

Embodiment 1: A Method of Preparing the Crystal Form A

[0104] Preparation method I: 20 mg of the compound as shown in formula I was weighed and added to a 4 mL glass bottle, and 50 times the volume of ethanol (EtOH) was added to the glass bottle, then the mixture was sonicated for 1 minute to obtain a suspension of the sample, and the suspension sample bottle was wrapped with an aluminum foil to shading and placed on a Labquaker rotator, and the mixture was equilibrated at room temperature (about 20 to 25° C.) with 360° rotation, and samples were taken at 10 days and 20 days, respectively, centrifuged, and characterized by XRPD after drying, the results were characterized as crystal form A.

[0105] Preparation method II: 20 mg of the compound as shown in formula I was weighed and added to a 10 mL glass bottle, then 130 times the volume of ethanol (EtOH) was added to the glass bottle; the sample was placed on a magnetic heating stirrer, and the water bath temperature was about 50° C., and the rotation speed was 200 rpm. The sample was heated to accelerate the dissolution, and the heat preservation was carried out for 15 min, and the solution was filtered while hot with a 0.45 μm filter membrane. The filtrate was transferred to a new glass bottle, and slowly cooled down to room temperature (20 to 25° C.) at a rate of 6° C./h. The solvent system of the precipitated solid was centrifuged and the solid was taken out, and after the solvent was evaporated to dryness, then the solid was characterized by XRPD, and the results were characterized as crystal form A.

[0106] Preparation method III (anti-solvent method): About 80 mg of the compound as shown in formula I sample was weighed and added to a 20 mL glass bottle, then 130 times the volume of good solvent ethanol (EtOH) was added to the bottle. The sample was placed on a magnetic heating stirrer, and the water bath temperature was about 50° C., and the rotation speed was 200 rpm. The sample was heated to accelerate the dissolution, and the heat preservation was carried out for 15 min, and the solution was filtered while hot with a 0.45 μm filter membrane. The filtrate was transferred to a new bottle, and 850 times the volume of the anti-solvent n-heptane was slowly added dropwise to each bottle under stirring, and the heat preservation was carried out for 10 min, then naturally cooled down. The solvent system of the precipitated solid was centrifuged and the solid was taken out, and after the solvent was evaporated to dryness, then the solid was characterized by XRPD, and the results were characterized as crystal form A.

Embodiment 2: A Method of Preparing the Crystal Form B

[0107] Preparation method I: 200 mg of the compound as shown in formula I was weighed and added to a bottle, then 10 times the volume of ethanol and 40 times the volume of water were added to the bottle, and the mixture was magnetic stirred and slurried for one day at room temperature. The solution was centrifuged, and the solid was collected, and dried at 40° C. for 4 hours; the solid after drying was characterized, and the results were characterized as crystal form B.

[0108] Preparation method II: 100 mg of the compound as shown in formula I was weighed and added to a glass bottle, then 30 times the volume of water was added to the glass bottle, and the mixture was magnetic stirred and slurried for 17 days at room temperature. The solution was filtered under reduced pressure, and dried at 40° C. for 4 hours, and then the solid was characterized by XRPD, and the results were characterized as crystal form B.

[0109] Preparation method III (anti-solvent method): About 80 mg of the compound as shown in formula I sample was respectively weighed and added to a 20 mL glass bottle. According to the following table, an appropriate volume of good solvent (see table 4 for specific volume) was respectively added to the bottle; the sample bottle was placed on a magnetic heating stirrer, and the water bath temperature was about 50° C., and the rotation speed was 200 rpm. The temperature in the water bath was maintained to promote sample dissolution, and the heat preservation was carried out for 15 min, and the solution was filtered while hot with a 0.45 μm filter membrane. The filtrate was transferred to a new bottle, and different anti-solvents were slowly added dropwise to each bottle in turn under stirring, and the heat preservation was carried out for 10 min, then the mixture was naturally cooled down. The solvent system of the precipitated solid was centrifuged and the solid was taken out, and after the solvent was evaporated to dryness, then the solid was characterized by XRPD, and the results were characterized as crystal form B.

TABLE-US-00004 TABLE 4 Good solvent/ Volume Crystal Number anti-solvent multiple Remark form 1 Ethanol/water 130/130 Magnetic Crystal stirring at room form B temperature for 8 days 2 Tetrahydro-  90/245 Magnetic Crystal furan/water stirring at room form B temperature for 7 days

Embodiment 3: A Method of Preparing the Crystal Form C

[0110] Preparation method I: 200 mg of the compound as shown in formula I sample was weighed, then 20 times the volume of acetone and 2.5 times the volume of water were added thereto. The mixture was magnetic stirred and slurried for one day at room temperature, and then the solution was centrifuged, and the solid was collected, and dried at 70° C. for 4 hours. The solid after drying was characterized by XRPD, and the results were characterized as crystal form C.

[0111] Preparation method II: 200 mg of the compound as shown in formula I and 25 times the volume of isopropanol (IPA) were magnetic stirred and slurried for 7 days at room temperature, filtered under reduced pressure, and the solid was dried at 70° C. for 4 hours, and then the solid sample was characterized by XRPD, and the results were characterized as crystal form C.

[0112] Preparation method III: 20 mg of the compound as shown in formula I was weighed and added to a 4 mL glass bottle, and 50 times of the volume of N,N-dimethylacetamide (DMA) was added to the glass bottle, then the mixture was sonicated for 1 minute to obtain a suspension of the sample, and the suspension sample bottle was wrapped with an aluminum foil to shading and placed on a Labquaker rotator, and the mixture was equilibrated at room temperature (about 20 to 25° C.) with 360° rotation, and samples were taken at 10 days and 20 days, respectively, centrifuged, and characterized by XRPD after drying, the results were characterized as crystal form C.

[0113] The crystal forms A, B and C of the compound were subjected to structure determination, crystal form study and the like by X-ray powder diffraction (XRPD) pattern, differential scanning calorimetry (DSC) analysis, thermogravimetric analysis (TGA) or dynamic vapor sorption (DVS), etc.

Embodiment 4: Characterization of Crystal Form A by Polarized Light Microscope (as shown in FIG. 1)

[0114] A small amount of the crystal form A sample of the compound as shown in formula I was taken and placed on a glass slide with a scale, and an appropriate amount of liquid paraffin was added to disperse, covered with a cover glass, and placed under a microscope with a 10-fold objective lens to observe the shape and size of the particles and crystal form properties. The birefringence properties and crystal habit of the samples were displayed using crossed polarizers and photographed with a digital camera.

[0115] The results showed that the sample had obvious birefringence phenomenon under the polarized light microscope, and the sample was granular and rod-shaped, and the particle size was 10 to 100

Embodiment 5: Differential Scanning Calorimetry (DSC) Analysis of the Crystal Form A (as shown in FIG. 2)

[0116] 3.1820 mg of the crystal form A sample of the compound as shown in formula I was weighed, and placed in an unsealed aluminum tray; the sample was equilibrated at 25° C. in a nitrogen flow (50 mL/min), and then heated from 25° C. to 300° C. at a heating rate of 10° C./min, the results are detailed in table 5.

TABLE-US-00005 TABLE 5 Initial Maximum Peak temperature temperature area Sample (° C.) (° C.) (J/g) Crystal form A 246.01 246.77 118.0

Embodiment 6: Thermogravimetric Analysis (TGA) of the Crystal Form A (as shown in FIG. 3)

[0117] 15.3240 mg of the crystal form A sample of the compound as shown in formula I was weighed, and placed in a platinum sample tray; in a nitrogen flow (60 mL/min), the sample was heated from 25° C. to 300° C. at a heating rate of 10° C./min, and the sample was heated from 26.76° C. to 119.97° C., and the weight loss was only 0.1447%, indicating that the sample was almost free of water or solvent, the results are detailed in table 6.

TABLE-US-00006 TABLE 6 Initial End temperature temperature Weight Sample (° C.) (° C.) loss (%) Crystal form A 26.76 119.97 0.1447 26.76 246.16 4.797

Embodiment 7: X-ray Powder Diffraction (XRPD) Analysis of the Crystal Form A (as shown in FIG. 4)

[0118] The light source was CuK, the X-ray intensity was 40 KV/40 mA, the scanning mode was Theta-Theta, the scanning angle range was 4° to 40°, the step size was 0.05°, and the scanning speed was 0.5 second/step, and the results are detailed in table 7.

TABLE-US-00007 TABLE 7 2θ Relative Number (2θ ± 0.2°) intensity (%) 1 3.979 25.2 2 4.991 14.3 3 7.113 1.5 4 8.135 1.5 5 9.923 36.3 6 10.883 27.0 7 11.613 6.6 8 14.251 13.2 9 14.866 4.4 10 16.210 18.0 11 17.357 100.0 12 18.607 25.0 13 19.294 26.3 14 19.594 11.4 15 20.792 11.2 16 21.272 6.8 17 24.437 6.7 18 25.257 5.9 19 26.229 4.4 20 27.870 3.7 21 28.631 1.9 22 29.126 5.2 23 29.943 1.5

Embodiment 8: Dynamic Vapor Sorption (DVS) Analysis of the Crystal Form A (as shown in FIG. 5)

[0119] An appropriate amount of the crystal form A sample of the compound as shown in formula I was weighed and dried at 25° C. and 0% RH for 60 min. The hygroscopic characteristics of the sample were tested when the humidity changed from 0% RH to 95% RH, and the dehumidifying characteristics of the sample were tested when the humidity changed from 95% RH to 0% RH. The humidity change in each step size was 5% RH, the equilibrium standard was that the weight change rate within 10 min was less than 0.01%/min, and the longest equilibrium time was 2 hours. The DVS results showed that the crystal form A had a hygroscopic weight gain of 0.310% at 25° C. and 80% RH humidity, and had a hygroscopic weight gain of 0.409% at 95% RH humidity, indicating that the sample was slightly hygroscopic.

Embodiment 9: X-ray Powder Diffraction (XRPD) Analysis of the Crystal Form B (as shown in FIG. 6)

[0120] The light source was CuK, the X-ray intensity was 40 KV/40 mA, the scanning mode was Theta-Theta, the scanning angle range was 4° to 40°, the step size was 0.05°, and the scanning speed was 0.5 second/step, and the results are detailed in table 8.

TABLE-US-00008 TABLE 8 2θ Relative Number (2θ ± 0.2°) intensity (%) 1 3.424 89.1 2 6.576 100.0 3 9.732 3.4 4 11.304 3.7 5 12.905 3.2 6 13.918 8.7 7 14.467 13.9 8 16.406 17.6 9 17.567 11.6 10 18.217 26.6 11 19.297 87.8 12 20.557 16.2 13 20.901 35.7 14 22.460 24.7 15 23.872 8.3 16 25.084 12.1 17 25.878 20.0 18 26.379 25.5 19 28.983 10.2 20 29.531 8.0 21 30.459 1.6 22 32.171 2.3 23 34.297 2.0 24 37.676 1.9 25 38.902 1.6

Embodiment 10: Differential Scanning Calorimetry (DSC) Analysis of the Crystal Form B (as shown in FIG. 7)

[0121] 1.5330 mg of the crystal form B sample of the compound as shown in formula I was weighed, and placed in an unsealed aluminum tray; the sample was equilibrated at 25° C. in a nitrogen flow (50 mL/min), and then heated from 25° C. to 300° C. at a heating rate of 10° C./min, and the results are detailed in table 9.

TABLE-US-00009 TABLE 9 Initial Maximum temperature temperature Peak Sample (° C.) (° C.) area (J/g) Crystal form B 49.24 67.36 90.47 80.86 85.42 1.479 98.17 104.30 4.510 241.15 242.84 93.73

Embodiment 11: Thermogravimetric Analysis (TGA) of the Crystal Form B (as shown in FIG. 8)

[0122] 1.7310 mg of the crystal form B sample of the compound as shown in formula I was weighed, and placed in a platinum sample tray; in a nitrogen flow (60 mL/min), the sample was heated from 25° C. to 300° C. at a heating rate of 10° C./min, and the initial sample was heated from 25.3° C. to 92.5° C., and the weight loss was 5.2%.

Embodiment 12: Dynamic Vapor Sorption (DVS) Analysis of the Crystal Form B (as shown in FIG. 9)

[0123] An appropriate amount of the crystal form B sample of the compound as shown in formula I was weighed and dried at 25° C. and 0% RH for 60 min. The hygroscopic characteristics of the sample were tested when the humidity changed from 0% RH to 95% RH, and the dehumidifying characteristics of the sample were tested when the humidity changed from 95% RH to 0% RH. The humidity change in each step size was 5% RH, the equilibrium standard was that the weight change rate within 5 min was less than 0.01%/min, and the longest equilibrium time was 2 hours. The results showed that the sample had a weight gain of 7.235% from 0% RH to 95% RH, indicating that the sample was hygroscopic.

Embodiment 13: X-ray powder diffraction (XRPD) analysis of the crystal form C (as shown in FIG. 10)

[0124] The light source was CuK, the X-ray intensity was 40 KV/40 mA, the scanning mode was Theta-Theta, the scanning angle range was 4° to 40°, the step size was 0.05°, and the scanning speed was 0.5 second/step, and the results are detailed in table 10.

TABLE-US-00010 TABLE 10 2θ Relative Number (2θ ± 0.2°) intensity (%) 1 6.250 100 2 6.979 15.3 3 8.779 38.6 4 12.635 21.3 5 13.720 32.6 6 15.285 3.1 7 16.525 24.1 8 17.696 5.5 9 18.458 73.0 10 19.302 59.0 11 20.852 15.4 12 22.345 19.1 13 24.772 14.3 14 25.230 11.5 15 25.996 4.1 16 27.285 13.2 17 28.303 4.1 18 28.829 3.7 19 29.699 2.3 20 30.703 5.6 21 33.133 3.1 22 34.655 1.9 23 36.829 2.4 24 37.967 3.1

Embodiment 14: Differential Scanning Calorimetry (DSC) analysis of the Crystal Form C (as shown in FIG. 11)

[0125] 1.550 mg of the crystal form C sample of the compound as shown in formula I was weighed, and placed in an unsealed aluminum tray; the sample was equilibrated at 25° C. in a nitrogen flow (50 mL/min), and then heated from 25° C. to 300° C. at a heating rate of 10° C./min, and the results are detailed in table 11.

TABLE-US-00011 TABLE 11 Initial Maximum temperature temperature Peak Sample (° C.) (° C.) area (J/g) Crystal form C 58.03 114.19 66.51 240.85 242.94 99.33

Embodiment 15: Thermogravimetric Analysis (TGA) of the Crystal Form C (as shown in FIG. 12)

[0126] 5.3570 mg of the crystal form C sample of the compound as shown in formula I was weighed, and placed in a platinum sample tray; in a nitrogen flow (60 mL/min), the sample was heated from 25° C. to 300° C. at a heating rate of 10° C./min; the sample was heated from 24.0° C. to 58.0° C., and the weight loss was 0.62%; the sample was heated from 58.0° C. to 162.3° C., and the weight loss was 2.5%. The sample was heated to 200° C., and the crystal form C was converted into the crystal form A, and the results are detailed in table 12.

TABLE-US-00012 TABLE 12 Initial End temperature temperature Weight Sample (° C.) (° C.) loss (%) Crystal form C 24.03 58.03 0.6172 58.03 162.33 2.517

Embodiment 16: Dynamic Vapor Sorption (DVS) Analysis of the Crystal Form C (as shown in FIG. 13)

[0127] An appropriate amount of the crystal form C sample of the compound as shown in formula I was weighed and dried at 25° C. and 0% RH for 60 min. The hygroscopic characteristics of the sample were tested when the humidity changed from 0% RH to 95% RH, and the dehumidifying characteristics of the sample were tested when the humidity changed from 95% RH to 0% RH. The humidity change in each step size was 5% RH, the equilibrium standard was that the weight change rate within 5 min was less than 0.01%/min, and the longest equilibrium time was 2 hours. The results showed that the sample had a weight gain of 4.767% from 0% RH to 95% RH, indicating that the sample was hygroscopic.

Embodiment 17: Comparison of Equilibrium Solubility and Dissolution Rate of Crystal Forms A, B and C

[0128] To further study whether the equilibrium solubility and dissolution rate of crystal forms A, B and C are different in pure water and physiological related media in vivo, the equilibrium solubility and dissolution rate of four crystal forms in water, simulated gastric fluid (SGF), fasted-state simulated intestinal fluid (FaSSIF) and fed state simulated intestinal fluid (FeSSIF) were tested respectively. About 20 mg of samples A, B, and C were weighed and added to a 4 mL glass bottle, respectively, and 3 mL of medium was added in turn, respectively; the mixture was sonicated for 15 seconds, put in a shaker at 37° C. with a rotation speed of 200 rpm, and about 1 mL of the suspension was taken at 0.5 hours, 2 hours and 24 hours, respectively, centrifuged with a rotation speed of 12000 rpm for 5 min; if necessary, the supernatant was diluted (diluent: methanol/water=9/1) to a suitable multiple, or directly measured the concentration by HPLC, and the solubility value of each sample was calculated. The solubility test results at each time point are shown in table 13:

TABLE-US-00013 TABLE 13 Time 0.5 hours 2 hours 24 hours Crystal form- Solubility Solubility Solubility medium (μg/mL) (μg/mL) (μg/mL) A-Water 1.5 6.0 6.1 B-Water 2.5 7.5 1.2 C-Water 1.7 6.5 1.8 A-SGF 0.4 0.3 0.1 B-SGF 0.5 0.1 0.2 C-SGF 0.3 0.2 0.5 A-FaSSIF 8.7 2.0 4.3 B-FaSSIF 13.3 8.8 8.6 C-FaSSIF 9.1 2.2 4.5 A-FaSSIF 182.8 176.6 170.4 B-FaSSIF 176.4 175.9 179.7 C-FaSSIF 181.7 174.3 166.6

[0129] Remarks: SGF: simulated gastric fluid; FaSSIF: fasted-state simulated intestinal fluid; FeSSIF: fed state simulated intestinal fluid

[0130] The solubility test results showed that there was no significant difference in the dissolution rate and solubility of the three crystal forms in water, SGF, FaSSIF and FeSSIF.

Embodiment 18: Solid-state stability experiment of crystal forms A, B and C

[0131] The samples of crystal forms A, B, and C were weighed and added to a 20 mL colorless transparent glass bottle, respectively, and the sample bottles were respectively placed under the corresponding influencing factors and acceleration conditions (the sample bottles under humidity conditions were sealed with tin foil and placed with holes, and the other samples were covered tightly and placed), taken out after being placed for 1 week and 2 weeks respectively, and the sample content and related substances under various conditions were detected by HPLC; according to the same method as above, samples were respectively weighed and placed under a corresponding influencing factor and accelerated condition, and were taken out after being placed for 2 weeks for observing the appearance and XRPD characterization of each sample, and the physical stability of each crystal form was examined, and the test methods are detailed in table 14 and table 15; at the same time, 2 samples of the crystal forms A, B, and C of the compound as shown in formula I were respectively weighed accurately into 20 mL colorless transparent glass bottles and covered tightly, placed in a −20° C. refrigerator, and taken out as a standard for HPLC analysis at 2 weeks, and the test results are detailed in table 16 and table 17.

TABLE-US-00014 TABLE 14 Content liquid chromatography method for solid stability investigation Chromatographic Waters Xbridge C18 column Mobile phase A: 100 % pure water; B: 100 % methanol Time Percentage Percentage Gradient elution (minutes) of phase A of phase B program 0 30 70 13 10 90 18 10 90 18.1 30 70 25 30 70 Flow rate  1.0 mL/min Column temperature  40° C. Detection 220 nm wavelength Running time  25 min Injection volume  10 μL Diluent Methanol: acetonitrile = 50:50 Sample  0.3 mg/mL concentration

TABLE-US-00015 TABLE 15 Related substances liquid chromatography method for solid stability investigation Chromatographic Waters Xbridge ™ C18 column (4.6 × 150 mm, 3.5 μm) Mobile phase A: 100 % pure water; B: 100 % methanol Gradient elution Time (minutes) program 0 40 60 5 40 60 30 20 80 60 20 80 Flow rate  1.0 mL/min Column temperature  40° C. Detection 220 nm wavelength Running time  60 min Injection volume  5 μL Diluent Methanol: acetonitrile = 50:50 Sample  1.5 mg/mL concentration

Embodiment 16: Solid-state Stability Results of Crystal Forms A, B and C-content and Related Substances

[0132]

TABLE-US-00016 Sample Content Total related name Condition Time (%) substances (%) Crystal form −20° C. 2 weeks / 0.00 A-Control Crystal form High temperature 1 week 100.69 0.02 A (60° C.) 2 weeks 104.03 0.03 High humidity 1 week 104.75 0.10 (92.5% RH) 2 weeks 104.25 0.12 illumination 1 week 96.10 3.94 (4500 LuX) 2 weeks 88.50 11.18 Acceleration 1 week 104.34 0.04 (40° C./75% RH) 2 weeks 104.13 0.03 Crystal form −20° C. 2 weeks / 0.00 B-Control Crystal form High temperature 1 week 100.70 0.00 B (60° C.) 2 weeks 100.15 0.00 High humidity 1 week 100.09 0.00 (92.5% RH) 2 weeks 100.83 0.00 illumination 1 week 100.02 0.39 (4500 LuX) 2 weeks 99.69 1.28 Acceleration 1 week 103.22 0.00 (40° C./75% RH) 2 weeks 101.60 0.00 Crystal form −20° C. 2 weeks 100.00 0.00 C-Control Crystal form High temperature 1 week 98.41 0.00 C (60° C.) 2 weeks 100.18 0.00 High humidity 1 week 101.55 0.00 (92.5% RH) 2 weeks 100.59 0.00 illumination 1 week 102.18 0.52 (4500 LuX) 2 weeks 98.64 1.56 Acceleration 1 week 100.06 0.00 (40° C./75% RH) 2 weeks 99.89 0.00

Embodiment 17: Solid-state Stability Results of Crystal Forms A, B and C-appearance and Crystal Forms

[0133]

TABLE-US-00017 Stability investigation Characterization results Sample name condition Appearance Crystal form Crystal form A- −20° C. Off-white Crystal form A Control Crystal form A High temperature (60° C.) Off-white Crystal form A Illumination (4500 Lux) Off-white Crystal form A High humidity Off-white Crystal form A (92.5% RH) Acceleration Off-white Crystal form A (40° C./75% RH) Crystal form B- −20° C. Off-white Crystal form B Control Crystal form B High temperature (60° C.) Off-white Mixed crystal form Illumination (4500 Lux) Off-white Crystal form B High humidity Off-white Crystal form B (92.5% RH) Acceleration Off-white Crystal form B (40° C./75% RH) Crystal form C- −20° C. Off-white Crystal form C Control Crystal form C High temperature (60° C.) Off-white Crystal form C Illumination (4500 Lux) Off-white Crystal form C High humidity Off-white Crystal form C (92.5% RH) Acceleration Off-white Crystal form C (40° C./75% RH)

[0134] Conclusion:

[0135] The samples of crystal forms A, B and C placed under four conditions of high temperature, high humidity, illumination and acceleration for 2 weeks had the same appearance as the initial samples, and they were all off-white; there was no significant difference between the crystal form A and C and the initial sample, and the physical stability was good, while the crystal form B was converted into other crystal form at a high temperature (60° C.) and the physical stability was poor.

[0136] Crystal forms A, B and C were significantly affected by illumination, and the total related substances increased by 11.18%, 1.28% and 1.56% respectively at 2 weeks, indicating that illumination has a significant effect on crystal forms A, B and C, and the crystal forms A, B and C are necessary to protect from light.

Comparative Embodiment 1: Preparation of Crystal Form V (Suspension Equilibrium Method)

[0137] 100 mg of the compound as shown in formula I sample was weighed and added to a bottle, then 20 times the volume of methanol was added to the bottle, and the mixture was magnetic stirred and slurried for 14 days at room temperature. The solution was centrifuged, and the solid was collected, and dried at 40° C. for 4 hours, and the solid after drying was characterized, and the crystal form was defined as a crystal form V; the XRPD pattern of the crystal form V is as shown in FIG. 14, and the DSC pattern of the crystal form V is as shown in FIG. 15; the crystal form V was heated to 85° C., and the crystal form V was converted into the crystal form A.

[0138] FIG. 16 is the TGA pattern of the crystal form V (as shown in FIG. 16, the sample has a weight loss of 3.690% when heated from 24.8° C. to 82.7° C.).

Comparative Embodiment 2: Preparation of Crystal Form VII (Solution Heating-Slow Cooling Method)

[0139] 100 mg of the compound as shown in formula I sample was weighed and added to a glass bottle, and 100 times the volume of ethyl acetate (EA) and 20 times the volume of methanol (MeOH) were added to the bottle, and the sample was placed on a magnetic heating stirrer. The mixture was magnetic stirred to fully dissolve at a water bath of 50° C., filtered and the filtrate was slowly cooled to room temperature at a rate of 6° C./h, overnight, filtered under reduced pressure, and the solid was dried at 70° C. for 4 hours. The solid after drying was characterized, and the crystal form was defined as a crystal form VII; the XRPD pattern of the crystal form VII is as shown in FIG. 17, the sample was heated to 200° C. and cooled to room temperature to characterize XRPD, and the result showed that the crystal form VII was converted into the crystal form A.

[0140] FIG. 18 is the DSC pattern of the crystal form VII (as shown in FIG. 18, the DSC heat flow curve shows that the initial melting point is 241.9° C., and there are several endothermic and exothermic peaks before 200° C.).

[0141] FIG. 19 is the TGA pattern of the crystal form VII (as shown in FIG. 19, the sample has a weight loss of 0.75% when heated from 26.2° C. to 112° C. and a weight loss of 1.69% when heated from 112° C. to 200° C.).

Comparative embodiment 3: Preparation of crystal form VIII (suspension equilibrium method)

[0142] 100 mg of the compound as shown in formula I sample was weighed and added to a bottle, then 30 times the volume of acetonitrile (ACN) (sample mass (g) x volume multiples) was added to the bottle, and the mixture was magnetic stirred and slurried for 2 days at room temperature, filtered under reduced pressure, and the solid was collected, and dried at 40° C. for 4 hours. The solid after drying was characterized, and the crystal form was defined as a crystal form VIII, and the XRPD pattern of the crystal form VIII is as shown in FIG. 20. The sample was heated to 180° C., and the crystal form VIII sample was converted into the crystal form A.

[0143] FIG. 21 is the DSC pattern of the crystal form VIII (as shown in FIG. 21, the DSC heat flow curve shows that the initial melting point is 239.0° C., and there is a broad endothermic peak before 100° C.).

[0144] FIG. 22 is the TGA pattern of the crystal form VIII (as shown in FIG. 22, the sample has a weight loss of 3.409% when heated from 26.1° C. to 85.2° C.).

Comparative embodiment 4: Preparation of crystal form IX (solution heating-slow cooling method)

[0145] 100 mg of the compound as shown in formula I sample was weighed and added to a glass bottle, and 30 times the volume of N,N-dimethylacetamide (DMA) was added to the bottle, and the sample was placed on a magnetic heating stirrer. The mixture was magnetic stirred to fully dissolve at a water bath of 50° C., filtered, and the filtrate was slowly cooled to room temperature at a rate of 6° C./h, filtered under reduced pressure, and the solid was dried at 70° C. for 4 hours. The solid after drying was characterized, and the crystal form was defined as a crystal form IX, and the XRPD pattern of the crystal form IX is as shown in FIG. 23. The sample was heated to 180° C., and the crystal form IX sample was converted into the crystal form A.

[0146] FIG. 24 is the DSC pattern of the crystal form IX (as shown in FIG. 24, the DSC heat flow curve shows that the initial melting point is 241.8° C., and there are several endothermic and exothermic peaks before 200° C.).

[0147] FIG. 25 is the TGA pattern of the crystal form IX (as shown in FIG. 25, the sample has a weight loss of 0.035% when heated from 27.4° C. to 95° C. and a weight loss of 0.841% when heated from 95° C. to 200° C.).