Co-crystal of Compound I Dihydrochloride and Preparation Method and Use Thereof

Abstract

Provided are co-crystals of Compound I dihydrochloride and preparation methods thereof, pharmaceutical compositions containing the co-crystals, and uses of the co-crystals for preparing cardiac muscle myosin agonist drugs and drugs for treating heart failure. Compared with prior arts, the provided co-crystals of Compound I dihydrochloride have one or more improved properties, which solve the problems of prior arts and are of great value to the optimization and development of the drugs.

Claims

1. A co-crystal of Compound I dihydrochloride with fumaric acid ##STR00002##

2. The co-crystal of Compound I dihydrochloride with fumaric acid according to claim 1, wherein the molar ratio of Compound I dihydrochloride and fumaric acid is 2:1.

3. The co-crystal of Compound I dihydrochloride with fumaric acid according to claim 1, wherein the X-ray powder diffraction pattern comprises characteristic peaks at 2theta values of 6.2°±0.2°, 17.4°±0.2° and 25.8°±0.2° using CuKα radiation.

4. The co-crystal of Compound I dihydrochloride with fumaric acid according to claim 3, wherein the X-ray powder diffraction pattern comprises one or two or three characteristic peaks at 2theta values of 12.6°±0.2°, 19.6°±0.2° and 23.5°±0.2° using CuKα radiation.

5. The co-crystal of Compound I dihydrochloride with fumaric acid according to claim 3, wherein the X-ray powder diffraction pattern comprises one or two or three characteristic peaks at 2theta values of 15.4°±0.2°, 21.1°±0.2° and 26.3°±0.2° using CuKα radiation.

6. A process for preparing the co-crystal of Compound I dihydrochloride with fumaric acid according to claim 1, wherein the process comprises: adding Compound I dihydrochloride solid and fumaric acid solid into a solvent mixture of a nitrile and water, stirring to obtain the co-crystal of Compound I dihydrochloride with fumaric acid.

7. The process according to claim 6, wherein the molar ratio of said Compound I dihydrochloride solid and fumaric acid solid is 1:3-2:1, said nitrile is acetonitrile, and volume ratio of acetonitrile and water in said solvent mixture is 9:1.

8. A co-crystal of Compound I dihydrochloride with tartaric acid ##STR00003##

9. The co-crystal of Compound I dihydrochloride with tartaric acid according to claim 8, wherein the molar ratio of Compound I dihydrochloride and tartaric acid is 1:1.

10. The co-crystal form of Compound I dihydrochloride with tartaric acid according to claim 8, wherein the X-ray powder diffraction pattern comprises characteristic peaks at 2theta values of 17.2°±0.2°, 20.2°±0.2° and 25.7°±0.2° using CuKα radiation.

11. The co-crystal of Compound I dihydrochloride with tartaric acid according to claim 10, wherein the X-ray powder diffraction pattern comprises one or two or three characteristic peaks at 2theta values of 19.4°±0.2°, 24.4°±0.2° and 30.6°±0.2° using CuKα radiation.

12. The co-crystal of Compound I dihydrochloride with tartaric acid according to claim 10, wherein the X-ray powder diffraction pattern comprises one or two or three characteristic peaks at 2theta values of 18.0°±0.2°, 14.7°±0.2° and 21.3°±0.2° using CuKα radiation.

13. A process for preparing the co-crystal of Compound I dihydrochloride with tartaric acid according to claim 8, wherein the process comprises: adding Compound I dihydrochloride solid and tartaric acid solid in an ester, slurring and separating to obtain the co-crystal of Compound I dihydrochloride with tartaric acid.

14. The process according to claim 13, wherein the molar ratio of said Compound I dihydrochloride solid and tartaric acid solid is 1:3-1:1, and said ester is ethyl acetate.

15. A pharmaceutical composition, wherein said pharmaceutical composition comprises a therapeutically effective amount of the co-crystal of Compound I dihydrochloride with fumaric acid according to claim 1 and pharmaceutically acceptable excipients.

16. A pharmaceutical composition, wherein said pharmaceutical composition comprises a therapeutically effective amount of the co-crystal of Compound I dihydrochloride with tartaric acid according to claim 8 and pharmaceutically acceptable excipients.

17. A method of agonizing cardiac muscle myosin, comprising administering to a subject in need thereof a therapeutically effective amount of the co-crystal of Compound I dihydrochloride with fumaric acid according to claim 1.

18. A method of agonizing cardiac muscle myosin, comprising administering to a subject in need thereof a therapeutically effective amount of the co-crystal of Compound I dihydrochloride with tartaric acid according to claim 8.

19. A method for treating heart failure, comprising administering to a subject in need thereof a therapeutically effective amount of the co-crystal of Compound I dihydrochloride with fumaric acid according to claim 1.

20. A method for treating heart failure, comprising administering to a subject in need thereof a therapeutically effective amount of the co-crystal of Compound I dihydrochloride with tartaric acid according to claim 8.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

[0063] FIG. 1 shows an XRPD pattern of Form CSI in Example 1

[0064] FIG. 2 shows a TGA curve of Form CSI in Example 1

[0065] FIG. 3 shows an XRPD pattern of Form CSI in Example 2

[0066] FIG. 4 shows a TGA curve of Form CSI in Example 2

[0067] FIG. 5 shows an XRPD pattern overlay of Form CSI before and after storage (from top to bottom: initial, stored at 25° C./60% RH (sealed) for six months, stored at 25° C./60% RH (open) for six months, stored at 40° C./75% RH (sealed) for six months, stored at 40° C./75% RH (open) for six months, stored at 60° C./75% RH (sealed) for one month).

[0068] FIG. 6 shows a DVS plot of Form CSI

[0069] FIG. 7 shows an XRPD pattern overlay of Form CSI before and after DVS test (top: before DVS, bottom: after DVS)

[0070] FIG. 8 shows an XRPD pattern of Form CSIII in Example 6

[0071] FIG. 9 shows a TGA curve of Form CSIII in Example 6

[0072] FIG. 10 shows an XRPD pattern of Form CSIII in Example 7

[0073] FIG. 11 shows a TGA curve of Form CSIII in Example 7

[0074] FIG. 12 shows a DSC curve of Form CSIII in Example 7

[0075] FIG. 13 shows an XRPD pattern overlay of Form CSIII before and after storage (from top to bottom: initial, stored at 25° C./60% RH (sealed with desiccant) for three months, stored at 25° C./60% RH (open) for three months, stored at 40° C./75% RH (sealed with desiccant) for three months, stored at 60° C./75% RH (sealed with desiccant) for three month).

[0076] FIG. 14 shows an XRPD pattern overlay of Form A before and after grinding (top:

[0077] before grinding, bottom: after grinding).

[0078] FIG. 15 shows an XRPD pattern overlay of Form CSI before and after grinding (top: before grinding, bottom: after grinding).

[0079] FIG. 16 shows an XRPD pattern overlay of Form CSIII before and after grinding (top: before grinding, bottom: after grinding).

[0080] FIG. 17 shows an XRPD pattern overlay of Form CSI tableting under different pressure (from top to bottom: 20 kN, 10 kN, 5 kN, 0 kN).

[0081] FIG. 18 shows an XRPD pattern overlay of Form CSIII tableting under different pressure (from top to bottom: 20 kN, 10 kN, 5 kN, 0 kN).

[0082] FIG. 19 shows an XRPD pattern overlay of Form CSI and Form CSI drug product (from top to bottom: excipients, Form CSI drug product, and Form CSI).

[0083] FIG. 20 shows an XRPD pattern overlay of Form CSIII and Form CSIII drug product (from top to bottom: excipients, Form CSIII drug product, and Form CSIII).

[0084] FIG. 21 shows an XRPD pattern overlay of Form CSI drug product stored under different conditions (from top to bottom: initial, stored under 40° C.±2° C./75%±5% RH (sealed with 1 g desiccant) for 3 months)

[0085] FIG. 22 shows an XRPD pattern overlay of Form CSIII drug product stored under different conditions (from top to bottom: initial, stored under 40° C.±2° C./75%±5% RH (sealed with 1 g desiccant) for 3 months)

[0086] FIG. 23 shows a dissolution curve of Form CSI drug product and Form A drug product in pH6.8 PBS.

DETAILED DESCRIPTION

[0087] 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 changes in the materials and methods can be accomplished without departing from the scope of the present disclosure.

[0088] The abbreviations used in the present disclosure are explained as follows

[0089] XRPD: X-ray Powder Diffraction

[0090] TGA: Thermo Gravimetric Analysis

[0091] DSC: Differential Scanning calorimetry

[0092] HPLC: High Performance Liquid Chromatography

[0093] IC: Ion Chromatography

[0094] .sup.1H NMR: Proton Nuclear Magnetic Resonance

[0095] DVS: Dynamic Vapor Sorption

[0096] Instruments and methods used for data collection

[0097] The X-ray powder diffraction patterns for the use of stability characterization of

[0098] Form CSI were acquired by a Bruker D8 DISCOVER X-ray powder diffractometer. The parameters of the X-ray powder diffraction method of the present disclosure are as follows:

[0099] X-Ray source: Cu, Kα

[0100] Kα1 (Å): 1.54060; Kα2 (Å): 1.54439

[0101] Kα2/Kα1 intensity ratio: 0.50

[0102] Voltage: 40 (kV)

[0103] Current: 40 (mA)

[0104] Scan range (2θ): from 4.0 degree to 40.0 degree

[0105] Except the samples tested by the Bruker D8 DISCOVER X-ray powder diffractometer, the other X-ray powder diffraction patterns 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:

[0106] X-Ray source: Cu, Kα

[0107] Kα1 (Å): 1.5406; Kα2 (Å): 1.54439

[0108] Kα2/Kα1 intensity ratio: 0.50

[0109] Voltage: 30 (kV)

[0110] Current: 10 (mA)

[0111] Scan range (2θ): from 3.0 degree to 40.0 degree

[0112] Thermo 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:

[0113] Heating rate: 10° C./min

[0114] Purge gas: nitrogen

[0115] 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:

[0116] Heating rate: 10° C./min

[0117] Purge gas: nitrogen

[0118] Dynamic Vapor Sorption (DVS) was measured via an SMS (Surface Measurement Systems Ltd.) intrinsic DVS instrument. Typical Parameters for DVS test are as follows:

[0119] Temperature: 25° C.

[0120] Gas and flow rate: nitrogen, 200 mL/min

[0121] Rate of mass change: 0.002%/min

[0122] RH range: 0% RH to 95% RH

[0123] 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.

[0124] The assay of Compound I in Form CSIII of the present disclosure is detected by

[0125] HPLC and the parameters are shown in Table 1.

TABLE-US-00001 TABLE 1 HPLC Agilent 1290 with DAD detector Column Agilent ZORBAX Eclipse Plus C18 Rapid Resolution HD, 2.1*50 mm, 1.8 μm Mobile Phase A: 0.05% H.sub.3PO.sub.4 aqueous solution (pH 6.8, triethylamine) B: Acetonitrile Time (min) % B Gradient 0.0 30.0 1.0 30.0 3.0 40.0 6.0 70.0 6.1 30.0 8.0 30.0 Run Time 8.0 min Post Time 0.0 min Speed 0.5 mL/min Injection Volume 1 μL Detection UV at 254 nm Wavelength Column Temperature 40° C. Sample Temperature Room temperature Diluent H.sub.2O

[0126] The assay of tartaric acid in Form CSIII of the present disclosure is detected by HPLC and the parameters are shown in Table 2.

TABLE-US-00002 TABLE 2 HPLC Agilent 1260 with VWD detector Column Ultimate LP-C18, 250*4.6 mm, 5 μm Mobile Phase A: ACN: H.sub.2O (pH = 3.0, H.sub.3PO.sub.4) = 5:95 B: ACN Time(min) % B Gradient 0.0 0.0 10.0 0.0 15.0 70.0 25.0 70.0 26.0 0.0 40.0 0.0 Run Time 40.0 min Post Time 0.0 min Speed 0.8 mL/min Injection Volume 2 μL Detection Wavelength UV at 210 nm Column Temperature 30° C. Sample Temperature Room temperature Diluent H.sub.2O

[0127] The assay of chloride ion in Form CSIII of the present disclosure is detected by IC, and the parameters are shown in Table 3.

TABLE-US-00003 TABLE 3 IC Thermo Fisher Dionex Aquion Column Thermo Dionex IonPac AS22, 4 × 250 mm, 6.0 μm Mobile Phase 4.5 mM Na.sub.2CO.sub.3/1.4 mM NaHCO.sub.3 Injection Volume 25 μL Flow rate 1.0 mL/min Conductivity Cell 35° C. Temperature Column Temperature: 30° C. Suppressor Current 31 mA Running Time 8 min

[0128] The parameters for related substance detection in the present disclosure are shown in Table 4.

TABLE-US-00004 TABLE 4 HPLC Agilent 1290 with DAD detector Column Agilent ZORBAX Eclipse Plus C18 Rapid Resolution HD, 2.1*50 mm, 1.8 μm Mobile Phase A: 0.05% H.sub.3PO.sub.4 aqueous solution (pH 6.8, triethylamine) B: Acetonitrile Time (min) % B Gradient 0.0 30.0 1.0 30.0 3.0 40.0 6.0 70.0 6.1 30.0 8.0 30.0 Run Time 8 min Post Time 0 min Speed 0.5 mL/min Injection Volume 2 μL Detection Wavelength UV, 254 nm Column Temperature 40° C. Sample Temperature Room temperature Diluent 80% acetonitrile aqueous solution

[0129] The parameters for formulation dissolution detection in the present disclosure are shown in Table 5.

TABLE-US-00005 TABLE 5 HPLC Agilent 1290 with DAD detector Column Agilent ZORBAX Eclipse Plus C18 Rapid Resolution HD, 2.1*50 mm, 1.8 μm Mobile Phase A: 0.05% H.sub.3PO.sub.4 aqueous solution (pH 6.8, triethylamine) B: Acetonitrile Time (min) % B Gradient 0.0 30.0 1.0 30.0 3.0 40.0 6.0 70.0 6.1 30.0 8.0 30.0 Run Time 8 min Post Time 0 min Speed 0.5 mL/min Injection Volume 2 μL Detection Wavelength UV, 254 nm Column Temperature 40° C. Sample Temperature Room temperature Diluent 80% acetonitrile aqueous solution

[0130] Unless otherwise specified, the following examples were conducted at room temperature. Said “room temperature” is not a specific temperature, but a temperature range of 10-30° C.

[0131] According to the present disclosure, Compound I dihydrochloride as a raw material is solid (crystalline and amorphous), semisolid, wax, oil, liquid form or solution. Preferably, Compound I dihydrochloride as a raw material is a solid.

[0132] Raw materials of Compound I dihydrochloride used in the following examples were prepared by known methods in prior arts, for example, the method disclosed in WO2014152270A1.

EXAMPLES

Example 1: Preparation of Form CSI

[0133] 98.7 mg of Compound I dihydrochloride, 44.5 mg of fumaric acid, and 5.0 mL of acetonitrile/water (9:1, v/v) were mixed and the obtained suspension was stirred at room temperature for 13 days. The solid was separated and dried under vacuum at 25° C. for 40 minutes. The obtained solid is Form CSI and the XRPD pattern of Form CSI is substantially as depicted in FIG. 1, and the XRPD data are listed in Table 6.

[0134] The TGA curve of Form CSI shows about 2.9% weight loss when heated to 130° C., which is substantially as depicted in FIG. 2.

[0135] The .sup.1H NMR results show that the molar ratio of Compound I dihydrochloride and fumaric acid in Form CSI is 2:1 and the specific data are: .sup.1H NMR (400 MHz, DMSO-d6) δ 11.13 (s, 1H), 10.94 (s, 1H), 9.35 (d, J=2.3 Hz, 1H), 8.92 (d, J=2.5 Hz, 1H), 8.19 (ddd, J=16.2, 8.4, 2.1 Hz, 2H), 7.78 (d, J=8.8 Hz, 1H), 7.48-7.34 (m, 1H), 7.26 (t, J=8.0 Hz, 1H), 6.63 (s, 1H), 4.39 (s, 2H), 2.65 (s, 3H).

TABLE-US-00006 TABLE 6 2θ (°) d spacing (Å) Relative Intensity % 6.18 14.30 25.47 11.10 7.97 2.97 12.60 7.02 13.10 13.70 6.46 13.53 14.47 6.12 3.44 15.35 5.77 10.35 15.82 5.60 1.97 16.76 5.29 15.75 17.42 5.09 29.76 18.21 4.87 10.04 18.62 4.76 13.88 19.63 4.52 20.90 20.03 4.43 8.54 20.82 4.27 14.01 21.10 4.21 16.57 21.71 4.09 12.25 22.21 4.00 11.74 22.62 3.93 7.60 23.52 3.78 24.00 24.20 3.68 13.54 24.77 3.59 37.26 25.35 3.51 12.74 25.78 3.46 100.00 26.31 3.39 17.25 27.88 3.20 11.11 28.67 3.11 13.16 29.18 3.06 8.59 30.06 2.97 8.21 31.01 2.88 10.27 31.28 2.86 22.34 32.46 2.76 6.26 33.53 2.67 1.85 34.17 2.62 2.53 34.68 2.59 5.54 36.32 2.47 3.18 38.39 2.34 3.20

Example 2: Preparation of Form CSI

[0136] 9.9 mg of Compound I dihydrochloride, 5.1 mg of fumaric acid, and 0.5 mL of acetonitrile/water (9:1, v/v) were mixed and the obtained suspension was stirred at room temperature for 21 days. The solid was separated to obtain a crystalline solid.

[0137] The crystalline solid was confirmed to be Form CSI, and the XRPD pattern of Form CSI is substantially as depicted in FIG. 3, and the XRPD data are listed in Table 7.

[0138] The TGA curve of Form CSI shows about 2.9% weight loss when heated to 130° C., which is substantially as depicted in FIG. 4.

TABLE-US-00007 TABLE 7 2θ (°) d spacing (Å) Relative Intensity % 6.17 14.33 38.36 11.10 7.97 3.74 12.59 7.03 9.48 13.70 6.46 12.22 15.35 5.77 10.25 16.66 5.32 12.47 17.42 5.09 31.38 18.21 4.87 8.55 18.61 4.77 17.93 19.63 4.52 20.48 20.04 4.43 6.73 20.82 4.27 14.23 21.10 4.21 9.88 21.72 4.09 10.56 22.21 4.00 8.93 22.61 3.93 10.11 23.53 3.78 15.05 24.21 3.68 11.52 24.77 3.59 24.47 24.89 3.58 22.76 25.35 3.51 8.57 25.78 3.46 100.00 26.31 3.39 13.45 27.05 3.30 3.16 27.89 3.20 8.02 28.66 3.11 8.31 29.18 3.06 6.53 30.08 2.97 3.32 31.03 2.88 5.13 31.28 2.86 19.75 31.89 2.81 5.40 32.46 2.76 3.99 34.69 2.59 2.36 35.06 2.56 2.29 36.40 2.47 1.22 38.38 2.35 1.19

Example 3: Preparation of Form CSI

[0139] 423.9 mg of Compound I dihydrochloride solid, 238.6 mg of fumaric acid and 10 mL of acetonitrile/water (9:1, v:v) were mixed, and stirred at room temperature for 1 day. Then 5 mL of acetonitrile/water (9:1, v:v) was added into the system and the system was further stirred for 1 day. The solid was separated and dried under vacuum at 25° C. for 50 minutes to obtain Form CSI.

Example 4: Stability of Form CSI

[0140] Approximately 5 mg of solid samples of Form CSI were stored under different conditions of 25° C./60% RH, 40° C./75% RH, and 60° C./75% RH. Crystalline form and chemical impurity were checked by XRPD and HPLC, respectively. The results are shown in Table 8 and FIG. 5.

TABLE-US-00008 TABLE 8 Condition Time Solid Form Purity (%) Initial — Form CSI 99.71 25° C./60% RH (sealed) 6 Months Form CSI 99.65 25° C./60% RH (open) 6 Months Form CSI 99.66 40° C./75% RH (sealed) 6 Months Form CSI 99.64 40° C./75% RH (open) 6 Months Form CSI 99.65 60° C./75% RH (sealed) 1 Month Form CSI 99.70

[0141] The results show that Form CSI is stable for at least six months at 25° C./60% RH and 40° C./75% RH. Form CSI has good stability under both long-term and accelerated conditions. Form CSI is stable for at least one month at 60° C./75% RH. Form CSI has good stability under more stress condition.

Example 5: Humidity Stability of Form CSI

[0142] DVS was applied to test the stability of Form CSI under different humidity with about 10 mg of samples. The weight change at each relative humidity were recorded in a humidity range of 0-95% RH. The results are shown in Table 9.

TABLE-US-00009 TABLE 9 Form Weight loss prior art Form A 2.7% (30%-5% RH) Form CSI 0.21% (30-0% RH) .sup.  

[0143] The weight loss of prior art Form A under 30%-5% RH is 2.7% and Form A converted to dehydrated state Form C under 5% RH. The weight loss of Form CSI of the present disclosure under 30%-0% RH is only 0.21%, which is much lower than that of prior art Form A, indicating that Form CSI of the present disclosure has less weight change in lower humidity, and has better stability under low humidity.

[0144] The DVS plot of Form CSI is shown in FIG. 6 and the XRPD pattern overlay of Form CSI before and after DVS test is shown in FIG. 7. The results show that the crystalline state of Form CSI remains unchanged after DVS, which indicates that Form CSI has good humidity stability.

Example 6: Preparation of Form CSIII

[0145] 98.5 mg of Compound I dihydrochloride, 63.8 mg of L-tartaric acid, and 5.0 mL of ethyl acetate were mixed and the obtained suspension was stirred at room temperature for 18 days. Then 5.0 mL of ethyl acetate was added into the system and the system was stirred at room temperature for 14 days. The solid was separated and dried at 50° C. for 2.5 hours. The obtained solid was confirmed to be Form CSIII, and the XRPD pattern of Form CSIII is substantially as depicted in FIG. 8, and the XRPD data are listed in Table 10.

[0146] The TGA curve of Form CSIII shows about 0.3% weight loss when heated to 100° C., which is substantially as depicted in FIG. 9.

[0147] The assay of Compound I, chloride ion and tartaric acid in Form CSIII was determined by HPLC and IC. The test results show that the molar ratio of chloride ion and Compound I in Form CSIII is 2:1, and the molar ratio of Compound I and tartaric acid is 1:1. The results are shown in Table 11.

TABLE-US-00010 TABLE 10 2θ (°) d spacing (Å) Relative Intensity % 5.56 15.91 6.57 6.65 13.29 3.48 8.17 10.82 1.33 10.33 8.57 13.55 11.12 7.96 4.28 12.06 7.34 9.31 13.29 6.66 5.60 14.69 6.03 43.08 15.28 5.80 18.86 16.42 5.40 52.75 17.19 5.16 52.44 17.99 4.93 46.16 19.37 4.58 41.35 20.16 4.40 60.65 21.33 4.16 47.06 22.07 4.03 12.41 23.33 3.81 45.52 23.68 3.76 27.60 24.42 3.65 48.43 25.70 3.47 100.00 28.07 3.18 18.38 28.69 3.11 26.69 30.59 2.92 74.23 32.05 2.79 10.19 33.03 2.71 7.52 33.71 2.66 12.68 35.66 2.52 6.68 36.34 2.47 5.32 37.62 2.39 2.48

TABLE-US-00011 TABLE 11 Chloride ion:Compound I Compound I:Tartaric acid 2.006:1 1.001:1

Example 7: Preparation of Form CSIII

[0148] 587.4 mg of Compound I dihydrochloride solid, 384.6 mg of L-tartaric acid and 20 mL of ethyl acetate were mixed and stirred at room temperature for 11 days. The solid was separated and dried under vacuum at 50° C. for 2.5 hours. The obtained dried solid was further mixed with 13 mL of ethyl acetate and stirred at room temperature for 1 day. The solid was separated and dried under vacuum at 40° C. for about 2 hours. The obtained solid was confirmed to be Form CSIII, and the XRPD pattern of Form CSIII is substantially as depicted in FIG. 10, and the XRPD data are listed in Table 12.

[0149] The TGA curve of Form CSIII shows about 0.8% weight loss when heated to 150° C., which is substantially as depicted in FIG. 11.

[0150] The DSC curve of Form CSIII is substantially as depicted in FIG. 12, which shows two endothermic peaks at around 197° C. and 209° C.

TABLE-US-00012 TABLE 12 2θ (°) d spacing (Å) Relative Intensity % 5.50 16.07 5.25 6.61 13.38 5.59 10.27 8.61 12.07 11.05 8.01 4.05 12.07 7.33 9.73 13.28 6.67 9.77 14.68 6.04 54.19 15.14 5.85 17.02 16.38 5.41 75.01 17.27 5.13 42.34 17.99 4.93 18.38 18.32 4.84 12.12 19.08 4.65 22.23 19.46 4.56 65.74 20.24 4.39 62.10 21.32 4.17 48.88 22.16 4.01 13.95 23.44 3.80 60.58 23.77 3.74 30.59 24.45 3.64 52.36 25.31 3.52 21.59 25.71 3.46 100.00 26.75 3.33 16.35 28.14 3.17 18.98 28.53 3.13 22.25 28.76 3.10 19.96 30.18 2.96 22.10 30.58 2.92 58.06 31.13 2.87 21.02 32.16 2.78 12.89 34.07 2.63 9.96 35.21 2.55 8.64 36.29 2.48 8.10 37.39 2.41 6.09 38.96 2.31 7.49

[0151] As disclosed in WO2014152270A1, the prior art Form A began losing weight at room temperature in the heating process, and there is a weight loss of about 2 to 5% in the range of about 100° C. to about 150° C., and when heated to 75-100° C., Form A converts to Form B. While Form CSIII of the present disclosure only has a mass change of 0.80% when heated to 150° C., and there is no thermal signal in DSC process before 150° C., indicating that Form CSIII has no crystal transformation before 150° C. and has better stability at higher temperature (below 150° C.), which is more beneficial for the stability of formulation processing and industrial production.

Example 8: Stability of Form CSIII

[0152] Approximately 5 mg of solid samples of Form CSIII were stored under different conditions of 25° C./60% RH, 40° C./75% RH, and 60° C./75% RH. Crystalline form and chemical impurity were checked by XRPD and HPLC, respectively. The results are shown in Table 13 and FIG. 13.

TABLE-US-00013 TABLE 13 Condition Time Solid Form Purity (%) Initial — Form CSIII 99.32 25° C./60% RH (sealed with 3 Months Form CSIII 99.31 desiccant) 25° C./60% RH (open) 3 Months Form CSIII 99.35 40° C./75% RH (sealed with 3 Months Form CSIII 99.32 desiccant) 60° C./75% RH (sealed with 3 Months Form CSIII 99.36 desiccant)

[0153] The results show that Form CSIII is stable for at least 3 months at 25° C./60% RH and 40° C./75% RH. Form CSIII has good stability under both long-term and accelerated conditions. Form CSIII is stable for at least 3 months at 60° C./75% RH. Form CSIII has good stability under more stress condition.

Example 9: Humidity Stability of Form CSIII

[0154] DVS was applied to test the stability of Form CSIII under different humidity with about 10 mg of samples. The weight change at each relative humidity were recorded in a humidity range of 0-95% RH.

[0155] The weight loss of prior art Form A under 30%-5% RH is 2.7% and it converted to Form C under 5% RH. The weight loss of Form CSIII of the present disclosure under 30%-0% RH is only 1.81%. The results indicate that Form CSIII of the present disclosure has less weight change in a lower humidity range and has better stability under low humidity.

Example 10: Compressibility of CSIII

[0156] A manual tablet press was used for compression. 60 mg of Form CSIII and prior art Form A were weighed and added into the dies of a φ6 mm round tooling, compressed at 10 KN manually, then stored at room temperature for 24 h until complete elastic recovery, diameter (D) and thickness (L) were tested with a caliper. Hardness (H) was tested with an intelligent tablet hardness tester. Tensile strength of the powder was calculated with the following formula: T=2H/πDL. Under a certain force, the greater the tensile strength, the better the compressibility. The results are presented in Table 14.

TABLE-US-00014 TABLE 14 Thickness Diameter Hardness Tensile strength Form (mm) (mm) (kgf) (MPa) Form A 1.67 6.00 3.38 2.11 Form CSIII 1.62 6.00 3.51 2.26

[0157] The results indicate that Form CSIII has better compressibility compared with prior art Form A.

Example 11: Mechanical Stability of Form CSI and Form CSIII

[0158] Solid sample of prior art Form A, Form CSI and Form CSIII of the present disclosure were separately grounded manually for 5 minutes in mortars. The XRPD patterns overlay before and after grinding is shown in FIG. 14, FIG. 15 and FIG. 16.

[0159] The results show that the crystallinity of prior art Form A decreases after grinding, while Form CSI and Form CSIII of the present disclosure have no crystal transformation and the crystallinity almost has no change after grinding, which indicates that Form CSI and Form III have better grinding stability when compared with Form A in the prior art.

[0160] A certain amount of Form CSI and Form CSIII were compressed into tablets under 5 kN, 10 kN, 20 kN pressure with suitable tableting die. Crystalline form before and after tableting were checked by XRPD. The results show that Form CSI and Form CSIII have no crystal transformation after tableting under different pressure. The XRPD pattern overlays are shown in FIG. 17 and FIG. 18.

Example 12: Adhesiveness of Form CSI

[0161] 30 mg of Form CSI, Form CSIII and prior art Form A were weighed and then added into the dies of φ8 mm round tooling, compressed at 10 KN and held for 30 s. The punch was weighed and the amount of material sticking to the punch was calculated. The compression was repeated twice and the maximum amount of material sticking to the punch during the compression were recorded. Detailed experimental results are shown in Table 15. Test results indicate that the adhesiveness of Form CSI and Form CSIII of the present disclosure is superior to the prior art Form A and the maximum amount is less than ⅕ of that of the prior art.

TABLE-US-00015 TABLE 15 Form Maximum amount (mg) Form A 2.83 Form CSI 0.48 Form CSIII 0.40

Example 13: Preparation of Form CSI and Form CSIII Drug Product

[0162] The formulation and preparation process of Form CSI and Form CSIII are shown in Table 16 and Table 17, respectively. The XRPD overlay of Form CSI and Form CSIII before and after formulation process are shown in FIG. 19 and FIG. 20. The results show that Form CSI and Form CSIII remain stable before and after the formulation process.

TABLE-US-00016 TABLE 16 Form Form CSI Form CSIII No. Component mg/unit % (w/w) mg/unit % (w/w) Function 1 Drug substance* 34.01 13.60 38.98 15.59 API 2 Fumaric acid 34.01 13.60 38.98 15.59 pH regulator 3 Microcrystalline 88.23 35.29 78.29 31.32 Filler Cellulose (PH102) 4 Lactose monohydrate 75.00 30.00 75.00 30.00 Filler (Armor Pharma 150 mesh) 5 Hydroxypropyl 5.00 2.00 5.00 2.00 Binder methylcellulose (EXF) 6 Croscarmellose 6.25 2.50 6.25 2.50 Disintegrant sodium Subtotal 242.50 97.00 242.50 97.00 N/A 7 Croscarmellose 6.25 2.50 6.25 2.50 Disintegrant sodium 8 Magnesium stearate 1.25 0.50 1.25 0.50 Lubricant (5712) Total 250 100 250 100 N/A *The sample weight is calculated with the consideration of the molecular weight and TGA weight loss of different APIs and each tablet corresponds to 25 mg compound I.

TABLE-US-00017 TABLE 17 Stage Procedure Pre-blending According to the formulation, materials No. 1-6 were weighed into an LDPE bag and blended for 2 mins. Simulation The mixture was pressed by a single punch manual of dry tablet press (type: ENERPAC; die: φ 20 mm round; granulation flake weight: 500 mg ± 100 mg; pressure: 5 ± 1 KN) and flakes were obtained. The flakes were pulverized and sieved through a 20-mesh sieve. Final blending Materials No. 7-8 were weighed and added into an LDPE bag together with the flakes after dry granulation and the mixture was blended for 2 mins. Tableting The mixture was tableted by a single punch manual tablet press (type: ENERPAC; die: φ9 mm round; tablet weight: 250 mg ± 10 mg; pressure: 7 ± 1 KN)

Example 14 Stability of Form CSI and Form CSIII in Drug Product

[0163] The drug products of Form CSI and Form CSIII prepared according to Example 13 were stored under 40° C./75% RH condition. The chemical impurity and crystalline form of the sample were tested by HPLC and XRPD, respectively. The stability results of the Form CSI and Form CSIII drug products are shown in Table 18.

TABLE-US-00018 TABLE 18 Sample Time Form Purity (% ) FIG. Form CSI tablet Initial Form CSI 99.69 FIG. 21 3 Months Form CSI 99.71 Form CSI tablet Initial Form CSIII 99.31 FIG. 22 3 Months Form CSIII 99.35 Packing Condition 35 cc HDPE bottle + 1 g desiccant

[0164] The results indicate that Form CSI and Form CSIII drug products can keep physically and chemically stable under 40° C.±2° C./75%±5% RH for at least 3 months and the chemical purity remains substantially unchanged.

Example 15: Dissolution of Form CSI Drug Product

[0165] Dissolution test was performed on Form CSI and prior art Form A drug product obtained from example 13. Dissolution method according to Chinese Pharmacopoeia 2020<0931>was used. The conditions are shown in Table 19.

TABLE-US-00019 TABLE 19 Dissolution tester Agilent 708DS Method Paddle Strength 25 mg Volume 900 mL Speed 50 rpm Temperature 37° C. Time 5, 10, 15, 20, 30, 45, 60 min Supplementary medium No

[0166] Dissolution results of Form CSI and prior art Form A drug products are presented in Table 20, the dissolution curves are shown in FIG. 23, which indicate that Form CSI drug product possesses better dissolution.

TABLE-US-00020 TABLE 20 Medium pH 6.8 PBS Time (min) Form A Form CSI 0 0.0 0.0 5 79.9 87.4 10 83.5 90.0 15 86.1 92.2 20 88.1 92.9 30 89.9 94.5 45 91.8 95.4 60 93.4 95.9

[0167] 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.