Tartrate of selective CDK9 inhibitor and crystal form thereof

Abstract

Disclosed are a tartrate of 3-(5-fluoro-4-(4-methyl-2-(methylamino)thiazol-5-yl) pyrimidin-2-ylamino)-benzenesulfonamide and a polymorph thereof, which are inhibitors of protein kinases, in particular cyclin-dependent kinase 9 (CDK9), and can be used to treat proliferative disorders, such as cancer, and other diseases related to protein kinase/CDK activity.

Claims

1. A 3-(5-fluoro-4-(4-methyl-2-(methylamino)thiazol-5-yl) pyrimidin-2-ylamino)-benzenesulfonamide tartrate having a structure as represented by Formula II: ##STR00003##

2. The tartrate according to claim 1 which is Crystal Form A of the tartrate, characterized in that it has a X-ray powder diffraction pattern with peaks located at positions with 2θ values of about 7.3, 9.6, 11.0, 15.3, 18.1, 18.9, 23.8, 24.5, 26.2, 26.7, and 27.1, a Raman spectrum with peaks located at positions with Raman shift values of about 1613 cm.sup.−1, 1597 cm.sup.−1, 1571 cm.sup.−1, 1543 cm.sup.−1, 1389 cm.sup.−1, 827 cm.sup.−1, and 543 cm.sup.−1, and a DSC thermogram with a sharp endothermic peak at 238.6° C.

3. The tartrate according to claim 1 which is Crystal Form B of the tartrate, characterized in that it has a X-ray powder diffraction pattern with peaks located at positions with 2θ values of about 8.0, 8.9, 9.5, 10.5, 14.8, 15.3, 16.1, 17.9, 18.9, 24.5, and 26.5, a Raman spectrum with peaks located at positions with Raman shift values of about 1612 cm.sup.−1, 1596 cm.sup.−1, 1569 cm.sup.−1, 1540 cm.sup.−1, 1519 cm.sup.−1, 1388 cm.sup.−1, 1286 cm.sup.−1, 1259 cm.sup.−1, 827 cm.sup.−1, and 543 cm.sup.−1, and a DSC thermogram with a sharp endothermic peak at 239.9° C.

4. A pharmaceutical formulation comprising 3-(5-fluoro-4-(4-methyl-2-(methylamino) thiazol-5-yl) pyrimidin-2-ylamino)-benzenesulfonamide tartrate and one or more pharmaceutically acceptable excipients.

5. A preparation method of the Crystal Form A of 3-(5-fluoro-4-(4-methyl-2-(methylamino)thiazol-5-yl) pyrimidin-2-ylamino)-benzenesulfonamide tartrate according to claim 2, comprising the following steps: (1) adding dimethyl sulfoxide to 3-(5-fluoro-4-(4-methyl-2-(methylamino)thiazol-5-yl)pyrimidin-2-ylamino)-benzenesulfonamide in an amount that is 4 to 8 times that of 3-(5-fluoro-4-(4-methyl-2-(methylamino)thiazol-5-yl)pyrimidin-2-ylamino)-benzenesulfonamide, heating to completely dissolve, and filtering while the liquid is hot; (2) adding a certain amount of tartaric acid and water to the first mixture, and reacting to obtain a second mixture; and (3) adding a certain amount of a water-miscible solvent to the second mixture, and reacting to obtain 3-(5-fluoro-4-(4-methyl-2-(methylamino)thiazol-5-yl) pyrimidin-2-ylamino)-benzenesulfonamide tartrate, which is the Crystal Form A of the tartrate.

6. The method according to claim 5, wherein the water-miscible solvent in step (3) is an alcohol.

7. The method according to claim 6, wherein the alcohol is ethanol.

8. The method according to claim 5, wherein the second mixture is obtained by mixing tartaric acid and 3-(5-fluoro-4-(4-methyl-2-(methylamino)thiazol-5-yl) pyrimidin-2-ylamino)-benzenesulfonamide at a molar ratio of 1.1 to 1.3:1.

9. A method of treating diseases or conditions caused by proliferative disorders, the method comprising administering to a patient in need thereof the 3-(5-fluoro-4-(4-methyl-2-(methylamino) thiazol-5-yl) pyrimidin-2-ylamino)-benzenesulfonamide tartrate according to claim 1.

10. The method according to claim 9, wherein the diseases or conditions caused by proliferative disorders are cancer.

11. The method according to claim 10, wherein the cancer is leukemia.

12. A use method of treating a disease or condition responsive to the inhibition of a protein kinase, the method comprising administering to a patient in need thereof the 3-(5-fluoro-4-(4-methyl-2-(methylamino) thiazol-5-yl) pyrimidin-2-ylamino)-benzenesulfonamide tartrate according to claim 1.

13. A use method of treating a disease or condition responsive to the inhibition of a protein kinase, the method comprising administering to a patient in need thereof the Crystal Form A of 3-(5-fluoro-4-(4-methyl-2-(methylamino) thiazol-5-yl) pyrimidin-2-ylamino)-benzenesulfonamide tartrate according to claim 2.

14. The method of claim 11, wherein the leukemia is acute myeloid leukemia.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

(1) Various features, advantages, and other applications of the present invention will be more obvious with reference to the description below and the accompanying drawings.

(2) FIG. 1 is an X-ray powder diffraction pattern of Crystal Form A of 3-(5-fluoro-4-(4-methyl-2-(methylamino) thiazol-5-yl) pyrimidin-2-ylamino)-benzenesulfonamide tartrate;

(3) FIG. 2 is an X-ray powder diffraction pattern of Crystal Form B of 3-(5-fluoro-4-(4-methyl-2-(methylamino) thiazol-5-yl) pyrimidin-2-ylamino)-benzenesulfonamide tartrate;

(4) FIG. 3 is an overlay of X-ray powder diffraction patterns of Crystal Form A and Crystal Form B of 3-(5-fluoro-4-(4-methyl-2-(methylamino) thiazol-5-yl) pyrimidin-2-ylamino)-benzenesulfonamide tartrate with 2θ values in a range from 0 to 40;

(5) FIG. 4 is an overlay of X-ray powder diffraction patterns of Crystal Form A and Crystal Form B of 3-(5-fluoro-4-(4-methyl-2-(methylamino) thiazol-5-yl) pyrimidin-2-ylamino)-benzenesulfonamide tartrate with 2θ values in a range from 0 to 20;

(6) FIG. 5 is an overlay of X-ray powder diffraction patterns of Crystal Form A and Crystal Form B of 3-(5-fluoro-4-(4-methyl-2-(methylamino) thiazol-5-yl) pyrimidin-2-ylamino)-benzenesulfonamide tartrate with 2θ values in a range from 20 to 40;

(7) FIG. 6 is an overlay of Raman spectra of Crystal Form A of 3-(5-fluoro-4-(4-methyl-2-(methylamino) thiazol-5-yl) pyrimidin-2-ylamino)-benzenesulfonamide tartrate with Raman shift values in a range from 0 cm.sup.−1 to 3000 cm.sup.−1;

(8) FIG. 7 is an overlay of Raman spectra of Crystal Form B of 3-(5-fluoro-4-(4-methyl-2-(methylamino) thiazol-5-yl) pyrimidin-2-ylamino)-benzenesulfonamide tartrate with Raman shift values in a range from 0 cm.sup.−1 to 3000 cm.sup.−1;

(9) FIG. 8 is an overlay of Raman spectra of Crystal Form A and Crystal Form B of 3-(5-fluoro-4-(4-methyl-2-(methylamino) thiazol-5-yl) pyrimidin-2-ylamino)-benzenesulfonamide tartrate with Raman shift values in a range from 0 cm.sup.−1 to 3000 cm.sup.−1;

(10) FIG. 9 is an overlay of Raman spectra of Crystal Form A and Crystal Form B of 3-(5-fluoro-4-(4-methyl-2-(methylamino) thiazol-5-yl) pyrimidin-2-ylamino)-benzenesulfonamide tartrate with Raman shift values in a range from 750 cm.sup.−1 to 1750 cm.sup.−1;

(11) FIG. 10 is an overlay of Raman spectra of Crystal Form A and Crystal Form B of 3-(5-fluoro-4-(4-methyl-2-(methylamino) thiazol-5-yl) pyrimidin-2-ylamino)-benzenesulfonamide tartrate with Raman shift values in a range from 100 cm.sup.−1 to 800 cm.sup.−1;

(12) FIG. 11 is a DSC thermogram of Crystal Form A of 3-(5-fluoro-4-(4-methyl-2-(methylamino) thiazol-5-yl) pyrimidin-2-ylamino)-benzenesulfonamide tartrate;

(13) FIG. 12 is a DSC thermogram of Crystal Form B of 3-(5-fluoro-4-(4-methyl-2-(methylamino) thiazol-5-yl) pyrimidin-2-ylamino)-benzenesulfonamide tartrate;

(14) FIG. 13 is an overlay of infrared spectra of Crystal Form A and Crystal Form B of 3-(5-fluoro-4-(4-methyl-2-(methylamino) thiazol-5-yl) pyrimidin-2-ylamino)-benzenesulfonamide tartrate with infrared shift values in a range from 500 cm.sup.−1 to 3500 cm.sup.−1;

(15) FIG. 14 is an overlay of infrared spectra of Crystal Form A and Crystal Form B of 3-(5-fluoro-4-(4-methyl-2-(methylamino) thiazol-5-yl) pyrimidin-2-ylamino)-benzenesulfonamide tartrate with infrared shift values in a range from 500 cm.sup.−1 to 2000 cm.sup.−1; and

(16) FIG. 15 is an overlay of infrared spectra of Crystal Form A and Crystal Form B of 3-(5-fluoro-4-(4-methyl-2-(methylamino) thiazol-5-yl) pyrimidin-2-ylamino)-benzenesulfonamide tartrate with infrared shift values in a range from 2500 cm.sup.−1 to 3000 cm.sup.−1.

DETAILED DESCRIPTION

Definitions

(17) The term “cancer” includes, but is not limited to, the following cancers: leukemia, breast cancer, ovarian cancer, cervical cancer, prostate cancer, testicular cancer, esophageal cancer, gastric cancer, skin cancer, lung cancer, bone cancer, colon cancer, pancreatic cancer, thyroid cancer, biliary tract cancer, throat cancer, lip cancer, tongue cancer, oral cancer, throat cancer, small intestine cancer, colon-rectal cancer, colorectal cancer, rectal cancer, brain and central nervous system cancer, malignant glioma, bladder cancer, liver cancer, kidney cancer, lymphoma, and the like.

(18) 3-(5-fluoro-4-(4-methyl-2-(methylamino) thiazol-5-yl) pyrimidin-2-ylamino)-benzenesulfonamide tartrate (Formula II) can be present in one or more polymorphic forms, including Form A and Form B. As described above, the polymorphic forms may be differentiated through X-ray powder diffraction, Raman spectroscopy, infrared spectroscopy, differential scanning calorimetry, or some combination of these characterization methods. The tartrate (Formula II) may be of high purity, i.e., containing at least 99% by weight of a particular polymorph, or may be a mixture of two polymorphs.

(19) FIG. 1 and FIG. 2 provide X-ray powder diffraction patterns of 3-(5-fluoro-4-(4-methyl-2-(methylamino) thiazol-5-yl) pyrimidin-2-ylamino)-benzenesulfonamide tartrate (Formula II), which define these polymorphic forms as Form A in FIG. 1 and Form B in FIG. 2. In order to facilitate comparison and reading, FIG. 3 is an overlay of X-ray powder diffraction patterns of Crystal Form A and Crystal Form B, and FIG. 4 and FIG. 5 are partially enlarged views of the overlay, respectively. Through comparison of the enlarged views, Polymorphic Form B is significantly different from Form A at 8.1, 10.6, 14.9, 16.1, etc. A person of ordinary skills in the field of polymorph identification is able to distinguish one crystal form from another crystal form by superimposing and comparing X-ray powder diffraction patterns and selecting a combination of characteristic peaks.

(20) The X-ray powder diffraction patterns shown in FIG. 1 to FIG. 5 are obtained on a Bruker D8 advance X-ray powder diffractometer using CuKα (40 kV, 40 mA) radiation. When the diffractometer is operated, the tube voltage and current are set to 40 kV and 40 mA, respectively, the distance from a sample to the detector: 30 cm, the scanning step: 0.1 s, and the scanning range: 3° to 40° (2θ).

(21) FIGS. 6-10 provide Raman spectra of 3-(5-fluoro-4-(4-methyl-2-(methylamino) thiazol-5-yl) pyrimidin-2-ylamino)-benzenesulfonamide tartrate (Formula II). FIG. 6 and FIG. 7 provide Raman spectra of Crystal Form A and Crystal Form B of the tartrate, respectively, wherein the Raman shift is from 0 cm.sup.−1 to 3000 cm.sup.−1. For convenience of comparison and reading, FIG. 8 is an overlay of Raman spectra of Crystal Form A and Crystal Form B. FIG. 9 and FIG. 10 are partially enlarged views of the overlay, respectively. Through comparison of the enlarged views, Polymorphic Form B is significantly different from Form A at 297 cm.sup.−1, 325 cm.sup.−1, 806 cm.sup.−1, and 1569 cm.sup.−1. A person of ordinary skills in the field of polymorph identification is able to distinguish one polymorphic form from another polymorphic form by choosing the above characteristic data or other characteristics.

(22) FIG. 11 and FIG. 12 respectively provide DSC thermograms of 3-(5-fluoro-4-(4-methyl-2-(methylamino) thiazol-5-yl) pyrimidin-2-ylamino)-benzenesulfonamide tartrate (Formula II) designated as Polymorphic Form A and Form B. The DSC data is obtained using a Perkin Elmer DSC 8500, temperature range: 50-280° C., scan rate: 10° C./min, and nitrogen flow rate: 20 mL/min. The DSC spectra show that Crystal Form A and Form B are heated to melt and decompose, and the two have similar melting points. Form A has a sharp endothermic peak at 238.6° C., and Form B has a sharp endothermic peak at 239.9° C.

(23) FIGS. 13-15 provide the infrared spectra of 3-(5-fluoro-4-(4-methyl-2-(methylamino) thiazol-5-yl) pyrimidin-2-ylamino)-benzenesulfonamide tartrate (Formula II). For convenience of comparison and reading, FIGS. 13-15 are all superimposed comparison diagrams of Crystal Form A and Form B. The infrared shift on FIG. 13 is 500 cm.sup.−1 to 3500 cm.sup.−1, the infrared shift on FIG. 14 is 500 cm.sup.−1 to 2000 cm.sup.−1, and the infrared shift on FIG. 15 is 2500 cm.sup.−1 to 3000 cm.sup.−1. It can be seen from the comparison of the overlay of infrared spectra that Polymorphic Form B is significantly different from Form A at 1641.1 cm.sup.−1, 3355.5 cm.sup.−1, and the like.

SPECIFIC IMPLEMENTATIONS

(24) The present invention will be further described in detail below in combination with examples, but is not limited thereto.

Example 1 Salt Formation Properties of 3-(5-fluoro-4-(4-methyl-2-(methylamino) Thiazol-5-yl) Pyrimidin-2-ylamino)-benzenesulfonamide Tartrate (with a Designation of LS007)

(25) 1.1 High-Throughput Screening of Salt Formation

(26) Based on the pKa value and solubilities at different pH of LS007, it can be determined that acids with pKa values of about 3 or lower may be used as acids for salt formation screening. Therefore, we selected 8 acids, including hydrochloric acid, sulfuric acid, aspartic acid, maleic acid, phosphoric acid, glutamic acid, tartaric acid, and fumaric acid.

(27) Dissolve the medicine and then add it to a 96-well plate, and determine the amount of counter ion to be added according to the molar mass of the added medicine and the quantity of the counter-ion functional groups. The heating time and temperature may be determined according to specific situation (typically 40° C. and 1 hour). In order to ensure a certain pressure in the flask during the reaction, the absolute tightness of a sample must be ensured during the mixing, vortex, and heating processes, and there must be at least a silicon resin liner throughout the entire process. The specific steps are as follows:

(28) 1) preparing a 0.02 M THF solution of the acids, wherein glutamic acid, sulfuric acid, and phosphoric acid are aqueous solutions;

(29) 2) preparing a 0.01 M THF/MeOH (1:1) solution of LS007;

(30) 3) adding 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 adding 1 mL solution of LS007, respectively; and

(31) 4) after vortex, reacting in a 40° C. oil bath for 1 hour, evaporating the organic solvent at room temperature, and finally reducing pressure for drying at 50° C.

(32) A comparison of Raman spectra shows that hydrochloric acid, sulfuric acid, phosphoric acid, maleic acid, tartaric acid, and fumaric acid all form a salt with LS007, while aspartic acid and glutamic acid do not form a salt.

(33) Perform scale-up experiments on the above six salts to determine solubilities of various salts in different pH buffers and deionized water, and compare with the free base. The results are listed in Table 1:

(34) TABLE-US-00001 TABLE 1 LS007 Hydrochloride Sulfate Phosphate Maleate Fumarate Tartrate Glycine- 134.1 904.4 141.8 1116.6 98.2 48.1 4302.3 HCl buffer (pH 2.0) Na.sub.2HPO.sub.4− 3.6 7.3 1.9 1.7 4.2 2.9 7.5 citric acid buffer (pH 4.5) Na.sub.2HPO.sub.4− 1.4 3.9 1.9 2.9 9.1 1.8 3.7 citric acid buffer (pH 6.8) Deionized 9.6 405.1 69.7 412.1 75.8 35.7 418.1 water

(35) Select the hydrochloride, phosphate, and tartrate with relatively good solubilities, and perform comprehensive solid-state characterization on the free base LS007, hydrochloride, phosphate, and tartrate. The comparison results are listed in Table 2:

(36) TABLE-US-00002 TABLE 2 Properties LS007 Hydrochloride Phosphate Tartrate Appearance Melting point TGA (decomposition temperature) Solubility 134.1 904.4 1116.6 4302.3 (25° C., 3.6 7.3 1.7 7.5 Glycine-HCl 1.4 3.9 2.9 3.7 buffer (pH 2.0) 9.6 405.1 412.1 418.1 Na.sub.2HPO.sub.4- citric acid buffer (pH 4.5) Na.sub.2HPO.sub.4- citric acid buffer (pH 6.8) Deionized water pH (saturated 3.06 2.70 2.50 3.03 aqueous solution 25° C.) Hygroscopicity 1.20 2.39 11.71 1.36 (DVS, 60% RH)

(37) The solubility results from HPLC testing show that the solubilities of the hydrochloride, phosphate, and tartrate in the pH 2.0 buffer and deionized water are significantly increased compared to those of the raw materials. Solubility: tartrate>phosphate>hydrochloride>free base.

(38) It can be seen from the DVS experiments on the Active Pharmaceutical Ingredients that LS007 has very low hygroscopicity, and hygroscopicity is increased after salt formation, wherein the phosphate has the highest hygroscopicity and absorbs 11.71% of water at 60% RH, followed by the hydrochloride, and the tartrate has the lowest hygroscopicity.

(39) The inventors are surprised to find that the tartrate has extraordinary performance in both solubility and hygroscopicity.

Example 2 Crystal Forms of 3-(5-fluoro-4-(4-methyl-2-(methylamino) Thiazol-5-yl) pyrimidin-2-ylamino)-benzenesulfonamide Tartrate

(40) For the polymorph issue of LS007 tartrate, this study has systematically screened possible crystal forms of compound LS007 tartrate by using different crystallization conditions and experimental approaches. Through nearly 300 crystallization experiments, it has been found that LS007 tartrate can exist in two different crystal forms, Crystal Form A and Crystal Form B, respectively. Further characterization has revealed that there is no significant difference in physicochemical properties between the different crystal forms. In the conversion experiments between the crystal forms, it has been found that Form A is a more stable crystal form, and Form B can be converted to Crystal Form A under certain conditions.

(41) (1) Form A

(42) Column-shaped crystal, melt and decompose the medicine, and the decomposition peak temperature is 236.8° C. It is non-hygroscopic (at 80% humidity, the hygroscopic weight gain is 0.22%). The amplitude of variation of humidity is low within a conventional storage humidity range. The physical and chemical properties are relatively ideal, the sample has the best crystallinity, the fluidity is greater than that of Form B, and the drug formation performance is better than that of Form B. Moreover, the equilibrium solubility is greater than that of Form B under various simulated in vivo conditions (pH=2.0, 4.6, 6.8).

(43) The specific preparation method of Crystal Form A is as follows:

(44) Add LS007 free base and dimethyl sulfoxide in a mass 6 times the mass of the free base into a four-necked flask, heat to completely dissolve (control the temperature <60° C.), and filter while hot; transfer the reaction mixture to a 10 L reaction flask, and add tartaric acid at 0.494 times (the weight of the free base, 1.3 equivalents) and water at 0.27 times (the weight of the free base), stir, heat to 60±2° C. and keep the temperature constant for half an hour; add anhydrous ethanol at 8.6 times (the weight of the free base), and keep the temperature constant at 60±2° C. for 4 hours; and lower the system temperature to 25±5° C., filter through suction (or centrifuge to dry), and wash the filter cake with an appropriate amount of anhydrous ethanol.

(45) Add the above solid and anhydrous ethanol 2-3 times the mass of the solid into the flask, stir at 80° C. for 1 hour, filter while hot, and dry the obtained filter cake with hot air at 80° C. to obtain a product as a yellow solid, which is the Crystal Form A of LS007 tartrate.

(46) (2) Form B

(47) Granular crystal, melt and decompose the medicine, and the decomposition peak temperature is 240.5° C. It is non-hygroscopic (at 80% humidity, the hygroscopic weight gain is 0.11%). The amplitude of variation of humidity is low within a conventional storage humidity range. Form B can be obtained when NM:H.sub.2O (1:1) is used as a solvent in a suspension experiment at 50° C.

(48) It can be seen from the XRPD overlay 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.

(49) All reference documents mentioned in the present invention are referenced in the present application, as if each reference document is individually referenced.