Tartrate salt of selective cdk9 inhibitor and its crystal form

[0053] Example 1 3-(5-Fluoro-4-(4-methyl-2-(methylamino)thiazol-5-yl)pyrimidin-2-ylamino)-benzenesulfonamide (code named LS007) salt-forming properties

[0054] 1.1 High-throughput screening for salt formation

[0055] Combined with the pKa value of LS007 and its solubility in different pH, it can be determined that acids with a pKa value of about 3 or less can be used as the acid for salt formation screening. Therefore, we chose 8 acids: hydrochloric acid, sulfuric acid, aspartic acid, maleic acid, phosphoric acid, glutamic acid, tartaric acid, and fumaric acid.

[0056] After dissolving the drug, add it to a 96-well plate, and determine the volume of counter ion to be added according to the molar amount of drug added and the number of counter ion functional groups. The heating time and temperature can be determined according to the specific circumstances (generally 40°C, 1 hour). In order to ensure a certain pressure during the reaction in the bottle, the absolute tightness of the sample should be ensured during the mixing, vortexing and heating process, and at least a silicone inner pad should be used during the whole process. The specific steps are as follows:

[0057] 1) Prepare a THF solution of 0.02M acid, where glutamic acid, sulfuric acid and phosphoric acid are aqueous solutions;

[0058] 2) Prepare 0.01M LS007 in THF/MeOH (1:1) solution;

[0059] 3) Add 1 mL of hydrochloric acid, 0.25 mL of sulfuric acid, 0.5 mL of aspartic acid, 0.5 mL of maleic acid, 0.5 mL of phosphoric acid, 0.5 mL of glutamic acid, 0.5 mL of tartaric acid, and 0.5 mL of fumaric acid, and then add 1 mL of LS007 each;

[0060] 4) After vortexing and reacting in an oil bath at 40°C for 1 hour, the organic solvent is evaporated at room temperature, and finally dried under reduced pressure at 50°C.

[0061] The comparison of Raman spectra shows that hydrochloric acid, sulfuric acid, phosphoric acid, maleic acid, tartaric acid and fumaric acid all form salts with LS007, while aspartic acid and glutamic acid do not form salts.

[0062] A scale-up experiment was performed on the above 6 kinds of salts to determine the solubility of various salts in different pH buffers and deionized water, and compared with free base. The results are shown in Table 1:

[0063] Table 1

[0064]

[0065] Choose the hydrochloride, phosphate and tartrate with better solubility, and do a comprehensive solid-state characterization of the free base LS007, hydrochloride, phosphate and tartrate. The comparison results are listed in Table 2:

[0066] Table 2

[0067]

[0068]

[0069] The solubility results of the HPLC test found that in pH 2.0 buffer and deionized water, the solubility of hydrochloride, phosphate and tartrate was significantly increased compared to the raw materials. Solubility: Tartrate> Phosphate> Hydrochloride> Free base.

[0070] It can be seen from the DVS experiment on the bulk drug that the hygroscopicity of LS007 is very small, and the hygroscopicity increases after salt formation. Phosphate has the strongest hygroscopicity and absorbs 11.71% of water at 60%RH; hydrochloride is the second; tartrate absorbs moisture. The least sex.

[0071] The inventor surprisingly discovered that tartrate has outstanding performance in terms of solubility and hygroscopicity.

[0072] Example 2 3-(5-Fluoro-4-(4-methyl-2-(methylamino)thiazol-5-yl)pyrimidin-2-ylamino)-benzenesulfonamide tartrate crystal form

[0073] In this study, aiming at the polymorphism of LS007 tartrate, we used different crystallization conditions and experimental methods to systematically screen the possible crystal forms of compound LS007 tartrate. Through nearly 300 crystallization experiments, it is found that LS007 tartrate can exist in two different crystal forms, namely crystal forms A and B. Further characterization found that there is no significant difference in physical and chemical properties between different crystal forms. In the conversion experiment between crystal forms, it was found that Form A is more stable crystal form, and Form B can be transformed into Form A crystal form under certain conditions.

[0074] (1)Form A

[0075] Columnar crystals, the drug melts and decomposes, the peak temperature of decomposition is 236.8℃. It does not attract humidity (at 80% humidity, the humidity gains 0.22%), and the humidity change is small within the normal storage humidity range. The physical and chemical properties are ideal, the sample crystallinity is the best, the fluidity is greater than that of Form B, the druggability is better than that of Form B, and the equilibrium solubility is greater than that of Form B under various simulated in vivo conditions (pH=2.0, 4.6, 6.8).

[0076] The specific preparation method of FormA crystal form is as follows:

[0077] In a four-necked flask, add LS007 free alkali, 6 times the mass of dimethyl sulfoxide, and heat to clear (control the temperature <60℃), filter while hot; transfer the reaction mixture to a 10L reaction flask, add 0.494 times (free base weight, 1.3 equivalents) of tartaric acid and 0.27 times (free base weight) of water, stir, and heat to 60±2°C Incubate for half an hour; add 8.6 times (free base weight) of absolute ethanol, and keep at 60±2℃ for 4 hours; the system temperature drops to 25±5℃, suction filtration (or centrifugal drying), filter cake with proper amount of absolute ethanol Rinse.

[0078] Add the above solids and 2-3 times the mass of absolute ethanol to the flask, stir at 80°C for 1 hour, filter while hot, and dry the filter cake with hot air at 80°C to obtain the yellow solid product of LS007 tartrate FormA crystal form.

[0079] (2)Form B

[0080] Granular crystals, the drug melts and decomposes, the peak temperature of decomposition is 240.5℃. It does not attract humidity (at 80% humidity, the humidity increases by 0.11%), and the humidity change is small within the normal storage humidity range. In the 50℃ suspension experiment, use NM: H 2 Form B can be obtained when O(1:1) is the solvent.

[0081] From the XRPD overlay, it can be seen that Form B is significantly different from Form A at 8.08°, 10.63°, 14.85°, 16.12°, 22.30°, 24.33°, 26.50°, etc.