CRYSTALLINE FORMS OF A FATTY ACID BILE ACID CONJUGATE, PREPARATION METHOD THEREOF AND USE THEREOF

Abstract

The present disclosure relates to novel crystalline forms of compound (I) and processes for preparation and use thereof. Crystalline forms CS3, CS2, CS5 and CS8 of compound (I) of the present disclosure have advantages in at least one aspect of solubility, stability, melting point, hygroscopicity, particle size, bioavailability, processability, purification effect, flowability, adhesiveness, stability in drug product, in vitro and in vivo dissolution, etc., which provides new and better choices for preparation of drug product containing compound (I) and has significant values for future drug development.

##STR00001##

Claims

1. A crystalline form CS3 of compound (I), wherein the X-ray powder diffraction pattern shows characteristic peaks at 2theta values of 20.5°±0.2°, 15.4°±0.2° and 22.3°±0.2° using CuKα radiation. ##STR00003##

2. The crystalline form CS3 according to claim 1, wherein the X-ray powder diffraction pattern shows one or two or three characteristic peaks at 2theta values of 7.8°±0.2°, 17.0°±0.2° and 17.9°±0.2° using CuKα radiation.

3. The crystalline form CS3 according to claim 1, wherein the X-ray powder diffraction pattern shows one or two or three characteristic peaks at 2theta values of 8.4°±0.2°, 5.1°±0.2° and 19.1°±0.2° using CuKα radiation.

4. A process for preparing crystalline form CS3 according to claim 1, wherein the process comprises: 1) Suspending solid of compound (I) into a solvent mixture comprising alcohols and water or alcohols and esters, then, stirring the solution at 0-80° C. to obtain crystalline form CS3; or 2) Adding solid of compound (I) into a solvent to obtain a solution of compound (I), and adding an anti-solvent slowly into the solution of compound (I) in solvent, wherein the solvent is halogenated hydrocarbons, and the anti-solvent is aromatic hydrocarbons; or adding the solution of compound (I) in solvent into an anti-solvent, wherein the solvent is ethers, and the anti-solvent is alkanes, stirring at room temperature to obtain crystalline form CS3.

5. The process for preparing crystalline form CS3 according to claim 4, wherein in method 1) said mixture of alcohols and water is mixture of isopropanol and water, said mixture of alcohols and esters is mixture of isopropanol and isopropyl acetate, in method 2) said halogenated hydrocarbon is chloroform, said aromatic hydrocarbon is toluene, said ether is tetrahydrofuran or 2-methyltetrahydrofuran, said alkane is n-heptane.

6. A crystalline form CS2 of compound (I), wherein the X-ray powder diffraction pattern shows characteristic peaks at 2theta values of 20.5°±0.2°, 16.8°±0.2° and 4.3°±0.2° using CuKα radiation.

7. The crystalline form CS2 according to claim 6, wherein the X-ray powder diffraction pattern shows one or two or three characteristic peaks at 2theta values of 8.4°±0.2°, 17.6°±0.2° and 14.9°±0.2° using CuKα radiation.

8. A process for preparing crystalline form CS2 according to claim 6, wherein the process comprises: 1) Mixing solid of compound (I) with methanol or a solvent mixture of ethanol and acetone, filtering after dissolved, storing the filtrate in a larger sealed device containing acetonitrile at 0-30° C. to obtain crystalline form CS2; or 2) Suspending solid of compound (I) into acetonitrile, then stirring the solution at 0-30° C. to obtain crystalline form CS2.

9. A crystalline form CS5 of compound (I), wherein the X-ray powder diffraction pattern shows characteristic peaks at 2theta values of 4.8°±0.2°, 9.7°±0.2° and 6.4°±0.2° using CuKα radiation.

10. The crystalline form CS5 according to claim 9, wherein the X-ray powder diffraction pattern shows one or two or three or four characteristic peaks at 2theta values of 20.9°±0.2°, 16.8°±0.2°, 23.3°±0.2° and 14.6°±0.2° using CuKα radiation.

11. A process for preparing crystalline form CS5 according to claim 9, wherein the process comprises: Adding solid of compound (I) into a solvent selected from alcohols to obtain a solution of compound (I), and adding acetonitrile slowly into the solution of compound (I), stirring at room temperature until solid precipitates, then, separating and drying to obtain crystalline form CS5.

12. The process for preparing crystalline form CS5 according to claim 11, wherein said alcohols are methanol, ethanol and isopropanol.

13. A crystalline form CS8 of compound (I), wherein the X-ray powder diffraction pattern shows characteristic peaks at 2theta values of 6.0°±0.2°, 15.5°±0.2° and 20.7°±0.2° using CuKα radiation.

14. The crystalline form CS8 according to claim 13, wherein the X-ray powder diffraction pattern shows one or two or three or four characteristic peaks at 2theta values of 21.8°±0.2°, 9.1°±0.2°, 19.9°±0.2° and 22.9°±0.2° using CuKα radiation.

15. A process for preparing crystalline form CS8 according to claim 13, wherein the process comprises: Storing crystalline form CS2 at room temperature for 7-30 days, heating the obtained sample to 95° C.±2° C. at a rate of 5-20° C./min with nitrogen purging, holding at 95° C.±2° C. for 1-10 min, and then cooling to room temperature at a rate of 5-20° C./min to obtain crystalline form CS8.

16. A pharmaceutical composition, wherein said pharmaceutical composition comprises a therapeutically effective amount of crystalline form CS3 according to claim 1, and pharmaceutically acceptable carriers, diluents or excipients.

17. A method for treating non-alcoholic steatohepatitis, gallstones, cholesterol gallstone, or atherosclerosis, comprising administering to a patient in need thereof a therapeutically effective amount of crystalline form CS3 according to claim 1.

18. (canceled)

19. A method for treating non-alcoholic steatohepatitis, gallstones, cholesterol gallstone, or atherosclerosis, comprising administering to a patient in need thereof a therapeutically effective amount of crystalline form CS2 according to claim 6.

20. A method for treating non-alcoholic steatohepatitis, gallstones, cholesterol gallstone, or atherosclerosis, comprising administering to a patient in need thereof a therapeutically effective amount of crystalline form CS5 according to claim 9.

21. A method for treating non-alcoholic steatohepatitis, gallstones, cholesterol gallstone, or atherosclerosis, comprising administering to a patient in need thereof a therapeutically effective amount of crystalline form CS8 according to claim 13.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

[0064] FIG. 1A shows an XRPD pattern of Form CS3 of the present disclosure.

[0065] FIG. 1B shows a TGA curve of Form CS3 of the present disclosure.

[0066] FIG. 1C shows a DSC curve of Form CS3 of the present disclosure.

[0067] FIG. 1D shows a .sup.1H NMR spectrum of Form CS3 of the present disclosure.

[0068] FIG. 1E shows an XRPD pattern overlay of Form CS3 of the present disclosure before and after being stored under 25° C./60% RH for one year, top: XRPD pattern before storage, bottom: XRPD pattern after storage.

[0069] FIG. 1F shows an XRPD pattern overlay of Form CS3 of the present disclosure before and after being stored under 40° C./75% RH for one year, top: XRPD pattern before storage, bottom: XRPD pattern after storage.

[0070] FIG. 1G shows a DVS plot of Form CS3 of the present disclosure.

[0071] FIG. 1H shows an XRPD pattern overlay of Form CS3 of the present disclosure before and after DVS test, top: XRPD pattern before test, bottom: XRPD pattern after test.

[0072] FIG. 1I shows a DVS plot of the prior art.

[0073] FIG. 1J shows a PSD diagram of Form CS3 of the present disclosure.

[0074] FIG. 1K shows a PSD diagram of the prior art.

[0075] FIG. 2A shows an XRPD pattern of Form CS2 of the present disclosure.

[0076] FIG. 2B shows a TGA curve of Form CS2 of the present disclosure.

[0077] FIG. 2C shows a DSC curve of Form CS2 of the present disclosure.

[0078] FIG. 2D shows an XRPD pattern overlay of Form CS2 of the present disclosure before and after DVS test, top: XRPD pattern before test, bottom: XRPD pattern after test.

[0079] FIG. 2E shows a PSD diagram of Form CS2 of the present disclosure.

[0080] FIG. 3A shows an XRPD pattern of Form CS5 of the present disclosure.

[0081] FIG. 3B shows a TGA curve of Form CS5 of the present disclosure.

[0082] FIG. 3C shows a DSC curve of Form CS5 of the present disclosure.

[0083] FIG. 3D shows a DVS plot of Form CS5 of the present disclosure.

[0084] FIG. 4A shows an XRPD pattern of Form CS8 of the present disclosure.

[0085] FIG. 4B shows a TGA curve of Form CS8 of the present disclosure.

[0086] FIG. 4C shows a DSC curve of Form CS8 of the present disclosure.

[0087] FIG. 4D shows an XRPD pattern overlay of Form CS8 of the present disclosure before and after being stored under 60° C./75% RH for 3 weeks, top: XRPD pattern before storage, bottom: XRPD pattern after storage.

[0088] FIG. 4E shows a DVS plot of Form CS8 of the present disclosure.

[0089] FIG. 4F shows an XRPD pattern overlay of Form CS8 of the present disclosure before and after DVS test, top: XRPD pattern before test, bottom: XRPD pattern after test.

DETAILED DESCRIPTION

[0090] The present disclosure is further illustrated by the following examples which describe the preparation and use of the crystalline forms of the present disclosure in detail. It is obvious to those skilled in the art that many changes in the materials and methods can be accomplished without departing from the scope of the present disclosure.

[0091] The abbreviations used in the present disclosure are explained as follows: [0092] XRPD: X-ray Powder Diffraction [0093] DSC: Differential Scanning calorimetry [0094] TGA: Thermal Gravimetric Analysis [0095] DVS: Dynamic Vapor Sorption [0096] .sup.1H NMR: Proton Nuclear Magnetic Resonance [0097] PSD: Particle Size Distribution

[0098] X-ray powder diffraction patterns in the present disclosure were acquired by a Bruker D2 PHASER X-ray powder diffractometer. The parameters of the X-ray powder diffraction method of the present disclosure are as follows: [0099] X-ray Reflection: Cu, Kα [0100] Kα1 (Å): 1.54060; Kα2 (Å): 1.54439 [0101] Kα2/Kα1 intensity ratio: 0.50 [0102] Voltage: 30 (kV) [0103] Current: 10 (mA) [0104] Scan range: from 3.0 degree to 40.0 degree

[0105] Differential scanning calorimetry (DSC) data in the present disclosure were acquired by a TA Q2000. The parameters of the DSC method of the present disclosure are as follows: [0106] Heating rate: 10° C./min [0107] Purge gas: nitrogen

[0108] Thermal gravimetric analysis (TGA) data in the present disclosure were acquired by a TA Q500. The parameters of the TGA method of the present disclosure are as follows: [0109] Heating rate: 10° C./min [0110] Purge gas: nitrogen

[0111] Proton nuclear magnetic resonance spectrum data (.sup.1H NMR) were collected from a Bruker Avance II DMX 400M HZ NMR spectrometer. 1-5 mg of sample was weighed and dissolved in 0.5 mL of deuterated dimethyl sulfoxide to obtain a solution with a concentration of 2-10 mg/mL.

[0112] The particle size distribution data in the present disclosure were acquired by an S3500 laser particle size analyzer of Microtrac. Microtrac S3500 is equipped with an SDC (Sample Delivery Controller). The test was carried out in wet mode, and the dispersion medium is Isopar G. The parameters are as follows:

TABLE-US-00001 Size distribution: Volume Run Time: 10 s Dispersion medium: Isopar G Particle coordinates: Standard Run Number: 3 Fluid refractive index: 1.42 Particle Transparency: Trans Residuals: Enabled Particle refractive index: 1.5 Flow rate: 60%* Particle shape: Irregular Filtration: Enabled Ultrasonication power: 30 W Ultrasonication time: 30 s *Flow rate 60% is 60% of 65 mL/s.

[0113] Unless otherwise specified, the following examples were conducted at room temperature. Solid of compound (I) used in the following examples were prepared by known methods in the prior art, the method disclosed in CN100386339C.

Example 1

[0114] Preparation of Form CS3 Example 1

[0115] As shown in Table 1A, certain amount of compound (I) was weighed into each of three glass vials followed by adding corresponding volume of solvent to form a suspension. These suspensions were stirred at RT, then centrifuged and dried. The obtained solid was collected and labeled as sample 1-a, 1-b, 1-c.

TABLE-US-00002 TABLE 1A Sample ID Mass (mg) Solvent (v/v) Volume (mL) 1-a 10.6 IPA/H.sub.2O 1:1 0.4 1-b 10.4 EtOH/EtOAc 1:9 0.5 1-c 9.1 IPA/IPAc 1:1 0.4

[0116] Sample 1-a, 1-b, 1-c were confirmed to be Form CS3 by XRPD. The XRPD pattern of sample 1-a is depicted in FIG. 1A, and the XRPD data are listed in Table 1B. The XRPD patterns of sample 1-b and 1-c are the same as or similar to that of sample 1-a.

TABLE-US-00003 TABLE 1B 2θ d spacing Intensity % 5.10 17.31 21.16 5.44 16.24 16.26 6.56 13.47 17.60 7.76 11.40 47.21 8.40 10.52 23.60 10.76 8.22 11.09 11.50 7.70 16.24 11.98 7.39 16.03 12.39 7.14 15.35 12.98 6.82 16.15 13.69 6.47 28.31 14.08 6.29 41.22 15.07 5.88 19.39 15.41 5.75 54.37 15.75 5.63 18.98 16.72 5.30 11.17 17.04 5.20 39.33 17.21 5.15 48.52 17.56 5.05 15.91 17.94 4.94 28.29 18.42 4.82 11.18 18.76 4.73 20.05 19.17 4.63 32.94 20.49 4.33 100.00 21.44 4.14 15.85 22.32 3.98 60.51 22.67 3.92 34.67 23.28 3.82 11.51 23.73 3.75 11.35 24.33 3.66 10.58 25.14 3.54 10.29 25.70 3.47 5.16 26.49 3.36 5.65

[0117] The TGA curve of Form CS3 shows about 0.3% weight loss when heated to 120° C., which is depicted in FIG. 1B.

[0118] The DSC curve of Form CS3 is depicted in FIG. 1C, which shows one endothermic peak at around 111° C. (onset temperature).

[0119] The .sup.1H NMR spectrum of Form CS3 is depicted in FIG. 1D, and the corresponding data are: .sup.1HNMR (400 MHz, DMSO) δ 11.95 (s, 1H), 7.53 (d, J=7.0 Hz, 1H), 4.12 (d, J=3.3 Hz, 1H), 4.05 (d, J=3.2 Hz, 1H), 3.83 (s, 1H), 3.77 (s, 1H), 3.61 (s, 1H), 2.26-1.93 (m, 6H), 1.89-1.03 (m, 53H), 1.04-0.79 (m, 10H), 0.59 (s, 3H). Except water, no other solvent residue was observed.

[0120] Preparation of Form CS3 Example 2:

[0121] 15.6 mg of compound (I) was dissolved into 0.5 mL of CHCl.sub.3. The solution was filtered and 2.0 mL of toluene was added into the filtrate. Then the clear solution was stirred until solid crystallized. The solid was collected by centrifugation, and the obtained solid was dried and labeled as sample 1-d. Sample 1-d was confirmed to be Form CS3.

[0122] Preparation of Form CS3 Example 3:

[0123] As shown in Table 1C, certain amount of compound (I) was dissolved in corresponding volume of solvent. After filtering, the filtrate was added into certain volume of anti-solvent. The suspension was stirred at RT until a large amount of solid precipitated. The solid was collected by centrifugation, and the obtained solid was dried and labeled as sample 1-e and 1-f. Sample 1-e and 1-f were confirmed to be Form CS3.

TABLE-US-00004 TABLE 1C Solvent Volume of Volume of Sample Mass Volume filtrate Anti- anti- solvent ID Method (mg) Solvent (mL) (mL) solvent (mL) 1-e Reverse 15.6 THF 0.5 0.2 n-Heptane 2.0 anti-solvent addition 1-f Reverse 15.4 2-MeTHF 1.0 0.4 n-Heptane 2.0 anti-solvent addition

[0124] Stability Study of Form CS3

[0125] As active pharmaceutical ingredient plays an important part in drug products, it is vital that the crystalline active pharmaceutical ingredient has good physical and chemical stability. Good physical stability avoids crystal transformation during the storage and formulation processes, thereby ensuring consistent and controllable quality of the drug substance and drug product.

[0126] Solid samples of Form CS3 were stored under different conditions of 25° C./60% RH and 40° C./75% RH in open dishes for one year. Crystalline form was checked by XRPD. The results are shown in Table 1D.

TABLE-US-00005 TABLE 1D Initial crystalline form Conditions Time Form change Form CS3 25° C./60% RH One No form change (FIG. 1E top) year observed (FIG. 1E bottom) Form CS3 40° C./75% RH One No form change (FIG. 1F top) year observed (FIG. 1F bottom)

[0127] The results show that Form CS3 has good stability and meets the requirements of the guidance of stability testing of drug substances and produces of China and/or American. Therefore, Form CS3 is suitable for drug development.

[0128] Hygroscopicity Study of Form CS3

[0129] Dynamic vapor sorption (DVS) was applied to test hygroscopicity of Form CS3 and the solid disclosed in CN100386339C with about 15 mg of samples. The results are listed in Table 1E. As shown in FIG. 1I, the solid disclosed in prior art is hygroscopic with 2.44% weight gain under 80% RH. While Form CS3 is non hygroscopic or almost non hygroscopic with 0.15% weight gain under 80% RH. DVS plot of test hygroscopicity of Form CS3 is depicted in FIG. 1G. What's more, the crystalline form of Form CS3 didn't change after DVS test, which is depicted in FIG. 1H.

TABLE-US-00006 TABLE 1E Weight gain under 80% The definition of Sample Relative Humidity hygroscopicity Form CS3 0.15% non hygroscopic or almost non hygroscopic Solid disclosed in 2.44% hygroscopic CN100386339C

[0130] Description and definition of hygroscopicity (Chinese Pharmacopoeia 2015 edition appendix XIX J Drug hygroscopic test guidelines, test at 25° C.+/−1° C., 80% RH.). [0131] deliquescent: Sufficient water is absorbed to form a liquid; [0132] very hygroscopic: Increase in mass is equal to or greater than 15 percent; [0133] hygroscopic: Increase in mass is less than 15 percent and equal to or greater than 2 percent; [0134] slightly hygroscopic: Increase in mass is less than 2 percent and equal to or greater than 0.2 percent. [0135] non hygroscopic or almost non hygroscopic: Increase in mass is less than 0.2 percent.

[0136] Particle Size Study of Form CS3

[0137] The particle size distribution of Form CS3 and the solid disclosed in CN100386339C were tested after ultrasonication for 30 seconds. The results are shown in Table 1F. The average particle size of Form CS3 is 26.3 μm, and D90 is 62.8 μm. The average particle size and D90 of the solid disclosed in prior art are 452.5 μm and 870.3 μm. Compared with prior art, Form CS3 has smaller particle size, which increases the specific surface area of the drug substance, improves the dissolution rate of drug, thereby facilitating drug absorption and further improving the bioavailability of the drug.

TABLE-US-00007 TABLE 1F Sample MV (μm) D10 (μm) D50 (μm) D90 (μm) Form CS3 26.3 2.0 12.3 62.8 Solid disclosed in 452.5 95.0 398.8 870.3 CN100386339C

[0138] Explanation of the abbreviations used in the present invention is as follows:

[0139] MV: Average particle diameter calculated by volume.

[0140] D10: the size in microns below which 10 percent of the particles reside on a volume basis.

[0141] D50: the size in microns below which 50 percent of the particles reside on a volume basis, also known as the median diameter.

[0142] D90: the size in microns below which 90 percent of the particles reside on a volume basis.

[0143] The particle size distribution diagrams of Form CS3 and the solid disclosed in prior art are shown in FIGS. 1J and 1K. As shown in FIG. 1J, Form CS3 has a narrow, nearly normal and uniform. Whereas, the solid disclosed in prior art has a large particle size and poor uniformity.

Example 2

[0144] Preparation of Form CS2 Example 1:

[0145] As shown in Table 2A, certain amount of compound (I) was dissolved in corresponding volume of solvent, and then filtered. The glass vial with filtrate was placed into a 20 mL glass vial with 5.0 mL of acetonitrile (ACN). The 20 mL glass vial was sealed and placed until solid crystallized. The obtained solids were labeled as sample 2-a˜2-c.

TABLE-US-00008 TABLE 2A Sample ID Mass (mg) Solvent (v/v) Volume (mL) 2-a 32.6 EtOH/Acetone 1:1 1.5 2-b 55.9 EtOH/Acetone 1:1 2.0 2-c 25.7 MeOH 1.4

[0146] Preparation of Form CS2 Example 2:

[0147] Approximately 5.3 mg of compound (I) was weighed into a 3 mL glass vial followed by adding 2.0 mL of ACN to form a suspension. The suspension was stirred at RT for 5 days and centrifuged. The obtained solids were labeled as sample 2-d.

[0148] Sample 2-a˜2-d were confirmed to be Form CS2 by XRPD. The XRPD pattern of sample 2-a is depicted in FIG. 2A, and the XRPD data are listed in Table 2B. The XRPD patterns of sample 2-b˜2-d are the same as or similar to that of sample 2-a.

TABLE-US-00009 TABLE 2B 2θ d spacing Intensity % 2.91 30.32 8.71 4.27 20.70 32.29 4.54 19.48 37.12 5.95 14.86 3.73 8.45 10.47 27.43 8.84 10.00 5.88 10.35 8.54 4.46 12.25 7.22 3.82 14.32 6.18 19.12 14.52 6.10 22.19 14.94 5.93 12.78 16.81 5.27 56.15 17.45 5.08 17.20 17.90 4.96 8.29 20.65 4.30 100.00 20.85 4.26 71.33 23.50 3.79 6.83 25.84 3.45 5.63 28.24 3.16 1.35 32.26 2.78 1.11 34.20 2.62 3.69 34.82 2.58 2.26

[0149] The TGA curve of Form CS2 shows about 2.9% weight loss when heated to 100° C., which is depicted in FIG. 2B. Form CS2 is a hydrate.

[0150] The DSC curve of Form CS2 is depicted in FIG. 2C, which shows one endothermic peak at around 83° C. (onset temperature).

[0151] Hygroscopicity Study of Form CS2

[0152] Dynamic vapor sorption (DVS) was applied to test hygroscopicity of Form CS2 with about 10 mg of samples. The results are listed in Table 2C and the XRPD patterns before and after DVS are shown in FIG. 2D. The result shows that weight gain of Form CS2 under 80% RH is 0.46%. Form CS2 is slightly hygroscopic according to the Chinese Pharmacopoeia 2015 edition appendix XIX J Drug hygroscopic test guidelines. As depicted in FIG. 1I, weight gain of the solid disclosed in prior art under 80% RH is 2.44%. The hygroscopicity of Form CS2 is lower than that of prior art. No form change of Form CS2 was observed before and after DVS test.

TABLE-US-00010 TABLE 2C Weight gain under 80% The definition of Sample Relative Humidity hygroscopicity Form CS2 0.46% slightly hygroscopic Solid disclosed in 2.44% hygroscopic CN100386339C

[0153] Particle Size Study of Form CS2

[0154] The particle size distribution of Form CS2 and the solid disclosed in CN100386339C were tested after ultrasonication for 30 seconds. The results are shown in Table 2D. An average particle size of Form CS2 is 41.9 μm, and D90 is 87.5 μm, while the average particle size and D90 of the solid disclosed in prior art are 452.5 μm and 870.3 μm. Compared with prior art, Form CS2 has smaller particle size, which increases the specific surface area of the drug substance, improves the dissolution rate of drug, thereby facilitating drug absorption and further improving the bioavailability of the drug.

TABLE-US-00011 TABLE 2D Sample MV (μm) D10 (μm) D50 (μm) D90 (μm) Form CS2 41.9 9.7 27.9 87.5 Solid disclosed in 452.5 95.0 398.8 870.3 CN100386339C

[0155] The particle size distribution diagrams of Form CS2 and the solid disclosed in prior art are shown in FIGS. 2E and 1K. As shown in FIG. 2E, Form CS2 has a narrow, nearly normal and uniform. Whereas, the solid disclosed in prior art has a large particle size and poor uniformity.

Example 3

[0156] Preparation of Form CS5 Example 1:

[0157] As shown in Table 3A, certain amount of compound (I) was dissolved in corresponding volume of solvent and then filtered. Then anti-solvent was added into the filtrate drop by drop until a large amount of solid precipitated. The suspension was then centrifuged and solid was obtained. The solid were dried at RT and labeled as sample 3-a˜3-c. Sample 3-a˜3-c were confirmed to be Form CS5 by XRPD. The XRPD pattern of sample 3-a is depicted in FIG. 3A, and the XRPD data are listed in Table 3B. The XRPD pattern of sample 3-b and 2-c are the same as or similar to that of sample 3-a.

TABLE-US-00012 TABLE 3A Volume of Volume of Volume of Sample Mass Solvent solvent filtrate Anti- anti-solvent ID (mg) ( v/v ) (mL) (mL) solvent (mL) 3-a 100.5 EtOH 2.0 2.0 ACN 10.0 3-b 92.2 MeOH 4.8 0.8 ACN 3.0 3-c 15.8 IPA 0.5 0.2 ACN 2.0

TABLE-US-00013 TABLE 3B 2θ d spacing Intensity % 3.11 28.41 35.83 3.34 26.43 36.32 4.76 18.57 100.00 5.12 17.26 56.39 6.43 13.74 25.69 6.88 12.85 14.22 7.90 11.19 9.13 9.71 9.11 26.62 10.43 8.48 15.50 11.31 7.82 6.40 12.11 7.31 4.84 13.96 6.34 7.31 14.56 6.08 11.44 16.34 5.42 10.45 16.82 5.27 15.50 17.86 4.97 10.06 19.60 4.53 4.80 20.51 4.33 15.06 20.92 4.25 25.84 21.72 4.09 6.44 23.30 3.82 12.46 28.28 3.16 2.88 33.16 2.70 2.22

[0158] The TGA curve of Form CS5 shows about 1.5% weight loss when heated to 90° C., which is depicted in FIG. 3B.

[0159] The DSC curve of Form CS5 is depicted in FIG. 3C, which shows one endothermic peak at around 91° C. (onset temperature).

[0160] Hygroscopicity Study of Form CS5

[0161] Dynamic vapor sorption (DVS) was applied to test hygroscopicity of Form CS5 with about 8 mg of samples. The results are listed in Table 3C. The result shows that weight gain of Form CS5 under 80% RH is 0.35%. Form CS5 is slightly hygroscopic according to the Chinese Pharmacopoeia 2015 edition appendix XIX J Drug hygroscopic test guidelines. As depicted in FIG. 1I, weight gain of the solid disclosed in prior art under 80% RH is 2.44%. The hygroscopicity of Form CS5 is lower than that of prior art. No form change of Form CS5 was observed before and after DVS test.

TABLE-US-00014 TABLE 3C Weight gain under 80% The definition of Sample Relative Humidity hygroscopicity Form CS5 0.35% slightly hygroscopic Solid disclosed in 2.44% hygroscopic CN100386339C

Example 4

[0162] Preparation of Form CS8 Example 1:

[0163] Form CS2 was stored at RT for one month. Then the sample was heated to 95° C. at a rate of 10° C./min with nitrogen purging, held at 95° C. for 2 min, and then cooled to RT at a rate of 5° C./min. The heating and cooling processes were conducted in a TGA equipment. The obtained sample was stored at 60° C./75% RH for one month to get Form CS8. The XRPD pattern of Form CS8 is depicted in FIG. 4A, and the XRPD data are listed in Table 4A. The TGA curve of Form CS8 shows about 0.9% weight loss when heated to 150° C., which is depicted in FIG. 4B. The DSC curve of Form CS8 is depicted in FIG. 4C, which shows one endothermic peak at around 104° C.

TABLE-US-00015 TABLE 4A 2θ d spacing Intensity % 5.96 14.83 100.00 7.97 11.10 8.80 9.19 9.62 24.41 10.52 8.41 6.32 13.33 6.64 8.28 13.92 6.36 13.11 14.52 6.10 20.11 15.50 5.72 82.15 15.76 5.62 46.17 16.67 5.32 21.95 17.18 5.16 32.62 18.00 4.93 17.41 18.32 4.84 10.15 19.90 4.46 29.70 20.60 4.31 78.93 21.71 4.09 66.13 22.80 3.90 28.42 22.96 3.87 27.23

[0164] Preparation of Form CS8 Example 2:

[0165] Form CS2 was stored at 25° C./60% RH for one week. The obtained sample was heated to 95° C. at a rate of 10° C./min with nitrogen purging, held at 95° C. for 2 min, and then cooled to RT at a rate of 10° C./min. The heating and cooling processes were conducted in a TGA equipment. Form CS8 was obtained and the XRPD data are listed in Table 4B.

TABLE-US-00016 TABLE 4B 2θ d spacing Intensity % 4.09 21.62 7.54 5.99 14.75 100.00 8.05 10.99 4.17 9.08 9.74 9.59 10.63 8.32 3.40 14.07 6.29 9.73 14.68 6.03 7.68 15.56 5.69 30.89 15.84 5.60 34.75 16.33 5.43 16.70 16.72 5.30 11.72 17.20 5.16 15.53 17.73 5.00 17.76 19.95 4.45 15.46 20.67 4.30 22.07 21.45 4.14 17.26 21.79 4.08 15.35 22.87 3.89 11.37

[0166] Stability Study of Form CS8

[0167] As active pharmaceutical ingredient plays an important part in drug products, it is vital that the crystalline active pharmaceutical ingredient has good physical and chemical stability. Good physical stability avoids crystal transformation during the storage and formulation processes, thereby ensuring consistent and controllable quality of the drug substance and drug product.

[0168] Form CS8 were stored in open dishes under 25° C./60% RH, 40° C./75% RH and 60° C./75% RH conditions for three weeks, and sampled for XRPD test. The results showed that no form change was observed, and the XRPD data of the sample before and after stored 60° C./75% RH for three weeks are shown in FIG. 4D as a representative.

[0169] Form CS8 is stable for at least three weeks under 25° C./60% RH, 40° C./75% RH and 60° C./75% RH. Further, Form CS8 is stable for at least three months. Further, Form CS8 is stable for at least six months. Further, Form CS8 is stable for at least one year. The results show that Form CS8 has good stability and is suitable for drug development.

[0170] Hygroscopicity Study of Form CS8

[0171] Dynamic vapor sorption (DVS) was applied to test hygroscopicity of Form CS8 with about 7 mg of sample. The results were listed in Table 4C and DVS curve is shown in FIG. 5E. The result showed that the weight gain of Form CS8 under 80% RH is 0.40%. Form CS8 is slightly hygroscopic according to the Chinese Pharmacopoeia 2015 edition appendix XIX J Drug hygroscopic test guidelines. As depicted in FIG. 1I, weight gain of the solid disclosed in prior art under 80% RH is 2.44%. The hygroscopicity of Form CS8 is lower than that of prior art. No form change of Form CS8 was observed before and after DVS test. The XRPD patterns before and after DVS were shown in FIG. 4F, top: the XRPD pattern before DVS; bottom: XRPD pattern after DVS.

TABLE-US-00017 TABLE 4C Weight gain under 80% The definition of Sample Relative Humidity hygroscopicity Form CS8 0.40% Slightly hygroscopic Solid disclosed in 2.44% Hygroscopic CN100386339C

[0172] The examples described above are only for illustrating the technical concepts and features of the present disclosure, and intended to make those skilled in the art being able to understand the present disclosure and thereby implement it, and should not be concluded to limit the protective scope of this disclosure. Any equivalent variations or modifications according to the spirit of the present disclosure should be covered by the protective scope of the present disclosure.