A crystal form of a JAK2 inhibitor and a preparation method thereof

Abstract

The invention belongs to the field of pharmaceutical chemistry technology, and in particular to A, B, C, D crystal forms and preparation methods of 2-{N-[4-(4-methyl)piperazine]phenyl}amino-5,6-dihydro-7-[3-(N-tert-butyl)sulfonamido]phenyl-pyrrolo[2,3-d]pyrimidine dihydrochloride. The four crystal forms A, B, C, and D provided by the present invention can be prepared by a variety of different conditions, and their crystallization process has a good purification effect, and has the advantageous features of stable process and easy operation. The preparation method of the crystal form is simple and low in cost, and 2-{N-[4-(4-methyl)piperazine]phenyl}amino-5,6-dihydro-7-[3-(N-tert-butyl)sulfonamido]phenyl-pyrrolo[2,3-d]pyrimidine dihydrochloride of different crystal forms with high purity, good solubility and good stability can be obtained.

Description

Technical Field

[0001] The present invention belongs to the field of medicinal chemistry, and specifically relates to A, B, C, and D crystal forms of 2-{N-[4-(4-methyl)piperazine]phenyl}amino-5,6-dihydro-7-[3-(N-tert-butyl)sulfonamido]phenyl-pyrrolo[2,3-d]pyrimidine dihydrochloride and a preparation method. Background Art

[0002] CN110305140A describes a compound 2-{N-[4-(4-methyl)piperazine]phenyl}amino-5,6-dihydro-7-[3-(N-tert-butyl)sulfonamido]phenyl-pyrrolo[2,3-d]pyrimidine, which as a free base shows highly selective JAK2 inhibition.

[0003] Given the importance of solid drug crystal forms and their stability in clinical treatment, further in-depth research on the polymorphic form of the compound 2-{N-[4-(4-methyl)piperazine]phenyl}amino-5,6-dihydro-7-[3-(N-tert-butyl)sulfonamido]phenyl-pyrrolo[2,3-d]pyrimidine dihydrochloride is of great significance for the development of drugs suitable for industrial production and with good biological activity. Summary of the Invention

[0004] In response to the problems of the solubility in water and oral bioavailability of the compound 2-{N-[4-(4-methyl)piperazine]phenyl}amino-5,6-dihydro-7-[3-(N-tert-butyl)sulfonamido]phenyl-pyrrolo[2,3-d]pyrimidine, the present inventors, after long-term efforts, discovered that preparing 2-{N-[4-(4-methyl)piperazine]phenyl}amino-5,6-dihydro-7-[3-(N-tert-butyl)sulfonamido]phenyl-pyrrolo[2,3-d]pyrimidine into a corresponding pharmaceutically acceptable salt can solve these problems.

[0005] The present invention first provides a pharmaceutically acceptable salt of 2-{N-[4-(4-methyl)piperazine]phenyl}amino-5,6-dihydro-7-[3-(N-tert-butyl)sulfonamido]phenyl-pyrrolo[2,3-d]pyrimidine, wherein the pharmaceutically acceptable salt is an inorganic salt or organic salt conventional in the art. In order to improve the physicochemical or biological properties of the drug, the free base (unless otherwise specified in the present invention, the free base refers to 2-{N-[4-(4-methyl)piperazine]phenyl}amino-5,6-dihydro-7-[3-(N-tert-butyl)sulfonamido]phenyl-pyrrolo[2,3-d]pyrimidine) can achieve faster dissolution and release in the body, which is beneficial to human absorption and the exertion of drug efficacy, and has greater clinical advantages. Furthermore, the inorganic salt is preferably dihydrochloride, sulfate, phosphate, or nitrate; and the organic salt is preferably methanesulfonate, citrate, oxalate, or succinate. The pharmaceutically acceptable salt is particularly preferably the dihydrochloride, which has advantages in solubility, stability and hygroscopicity over the free base and other salts.

[0006] The present invention further provides A, B, C, and D crystal forms of 2-{N-[4-(4-methyl)piperazine]phenyl}amino-5,6-dihydro-7-[3-(N-tert-butyl)sulfonamido]phenyl-pyrrolo[2,3-d]pyrimidine dihydrochloride and a preparation method.

[0007] The crystalline form A of the compound of formula (1), 2-{N-[4-(4-methyl)piperazine]phenyl}amino-5,6-dihydro-7-[3-(N-tert-butyl)sulfonamido]phenyl-pyrrolo[2,3-d]pyrimidine dihydrochloride, has characteristic peaks at 2θ of 5.989, 8.478, 12.288, 13.315, 16.985, and 17.476 in its X-ray powder diffraction pattern;

[0008]

[0009] Furthermore, the X-ray powder diffraction pattern of the A crystal form has characteristic peaks at diffraction angles 2θ of 3.105, 5.989, 8.478, 12.288, 13.315, 16.985, 17.476, 22.174, and 24.898.

[0010] Furthermore, the X-ray powder diffraction pattern of the A crystal form has characteristic peaks at diffraction angles 2θ of 3.105, 5.989, 6.765, 8.478, 12.288, 13.315, 16.432, 16.985, 17.476, 19.370, 22.174, 24.166, 24.898, and 30.243.

[0011] The present invention also provides a method for preparing the crystalline form A of the compound of formula (1), the method comprising:

[0012] (a) Add the compound of formula (1) to a solvent (I), and dissolve by stirring or heating to dissolve or slurry by heating. The solvent (I) is selected from at least one of water, ethanol, methanol, isopropanol, ethylene glycol, acetone, methyl tert-butyl ether, ethyl acetate, acetonitrile, tetrahydrofuran, and dioxane, preferably methanol, ethanol, or ethanol / water. The volume (ml) of the solvent (I) used is 1 to 60 times the weight (g) of the compound.

[0013] (b) Stirring crystallization.

[0014] (c) drying by heating, wherein the drying temperature is 40°C to 100°C, preferably 50°C to 90°C.

[0015] The present invention provides a crystal form B of a compound of formula (1). The X-ray powder diffraction pattern of the crystal form B has characteristic peaks at diffraction angles 2θ of 5.106, 12.765, 17.879, 22.419 and 24.214.

[0016] Furthermore, the X-ray powder diffraction pattern of the B crystal form has characteristic peaks at diffraction angles 2θ of 3.452, 5.106, 5.762, 6.339, 6.911, 8.900, 11.270, 12.765, 13.914, 17.347, 20.542, 22.419, and 24.214.

[0017] Furthermore, the X-ray powder diffraction pattern of the B crystal form has characteristic peaks at diffraction angles 2θ of 3.452, 5.106, 5.762, 5.957, 6.339, 6.911, 8.900, 10.406, 11.270, 11.924, 12.765, 13.914, 14.247, 17.347, 18.476, 20.542, 22.419, and 24.214.

[0018] The present invention also provides a method for preparing the crystalline form B of the compound of formula (1), the method comprising:

[0019] (a) Add the compound of formula (1) to solvent (II), stir and dissolve or heat and dissolve. The solvent (II) is selected from at least one of water, ethanol, methanol, isopropanol, and methyl tert-butyl ether, preferably methanol, ethanol / water, or methanol / methyl tert-butyl ether. The volume (ml) of the solvent (II) used is 1-60 times the weight (g) of the compound.

[0020] (b) Stirring crystallization.

[0021] (c) heating and drying, the drying temperature is 10°C-40°C, preferably 25°C-40°C.

[0022] The present invention provides a crystal form C of a compound of formula (1). The X-ray powder diffraction pattern of the crystal form C has characteristic peaks at diffraction angles 2θ of 5.743, 11.931, 13.097, 15.625, 17.066 and 18.429.

[0023] Furthermore, the X-ray powder diffraction pattern of the C crystal form has characteristic peaks at diffraction angles 2θ of 5.743, 6.262, 11.931, 12.561, 13.097, 13.967, 15.625, 17.066, 18.429, 18.882, 21.432, 24.496, and 24.931.

[0024] Furthermore, the X-ray powder diffraction pattern of the C crystal form has characteristic peaks at diffraction angles 2θ of 5.743, 6.262, 6.551, 11.931, 12.561, 13.097, 13.967, 15.625, 17.066, 18.429, 18.882, 19.180, 19.639, 21.432, 23.133, 24.496, and 24.931.

[0025] The present invention also provides a method for preparing Form C of the compound of formula (1), the method comprising:

[0026] (a) Add the compound of formula (1) to a solvent (IIIa), wherein the solvent (IIIa) is selected from at least one of ethanol, ethylene glycol, and acetone, preferably ethanol, and slurry the mixture at room temperature or under heating; or add the compound of formula (1) to a solvent (IIIb), wherein the solvent (IIIb) is selected from at least one of methanol and ethyl acetate, isopropyl acetate, or butyl acetate, preferably methanol / ethyl acetate, and dissolve the mixture by stirring or heating. The volume (ml) of the solvent (III) used is 1-60 times the weight (g) of the compound.

[0027] (b) Stirring crystallization.

[0028] (c) heating and drying, the drying temperature is 10°C-40°C, preferably 25°C-40°C.

[0029] The present invention provides a crystal form D of a compound of formula (1). The X-ray powder diffraction pattern of the crystal form D has characteristic peaks at diffraction angles 2θ of 6.733, 8.670, 11.931, 18.213 and 19.043.

[0030] Furthermore, the X-ray powder diffraction pattern of the D crystal form has characteristic peaks at diffraction angles 2θ of 6.733, 8.670, 11.931, 12.363, 18.213, 19.043, 23.033, and 24.813.

[0031] The present invention also provides a method for preparing the crystalline form D of the compound of formula (1), the method comprising:

[0032] (a) Add the compound of formula (1) to a solvent (IV), stir or heat to dissolve, and add an antisolvent dropwise for crystallization, wherein the solvent (IV) is selected from methanol, ethanol, ethylene glycol, and water, preferably methanol. The antisolvent is selected from acetonitrile. The volume ratio of the solvent (IV) to acetonitrile is 2:1-1:10, preferably 1:3, 1:4, or 1:5.

[0033] (b) heating and drying, the drying temperature is 20°C-80°C, preferably 25°C-50°C.

[0034] It should be understood that using different types of equipment or using different test conditions may give slightly different melting point readings. The exact value of the melting point of different crystalline forms will be affected by compound purity, sample weight, heating rate, particle size, and calibration and maintenance of the test equipment. The values ​​provided should not be regarded as absolute values.

[0035] It should be understood that using different equipment or testing conditions may produce slightly different XPRD patterns and peak values. The patterns, peak values, and relative intensities of diffraction peaks for different crystalline forms will be affected by compound purity, sample preparation, scanning speed, particle size, and calibration and maintenance of the testing equipment. The values ​​provided should not be considered absolute.

[0036] The "X-ray powder diffraction pattern or XPRD" mentioned in the present invention is obtained by Cu-Kα ray diffraction.

[0037] The "differential scanning calorimetry or DSC" mentioned in the present invention refers to measuring the temperature difference and heat flow difference between a sample and a reference object during the process of heating or maintaining the sample at a constant temperature, so as to characterize all physical and chemical changes related to thermal effects and obtain phase change information of the sample.

[0038] The diffraction angle 2θ described in the present invention is the Bragg angle, with the unit being degree, and the error range of the 2θ is ±0.2.

[0039] The beneficial effects of the present invention are as follows: the A, B, C, and D crystal forms of the compound of formula (1) provided by the present invention have advantages in stability, solubility, and formulation dissolution, are more suitable for drug development, meet the requirements of bioavailability and efficacy, can meet the pharmaceutical requirements of production, transportation, and storage, and the production process is stable, repeatable, and controllable, and can be adapted to industrial production.

[0040] The four crystal forms A, B, C, and D provided by the present invention can be prepared under a variety of different conditions. The crystallization process has good purification effects and has advantageous characteristics such as stable process and easy operation. The preparation method of the crystal form is simple and low-cost, and 2-{N-[4-(4-methyl)piperazine]phenyl}amino-5,6-dihydro-7-[3-(N-tert-butyl)sulfonamido]phenyl-pyrrolo[2,3-d]pyrimidine dihydrochloride in different crystal forms with high purity, good solubility, and good stability can be obtained. BRIEF DESCRIPTION OF THE DRAWINGS

[0041] Figure 1 XPRD pattern of Form A of the compound of formula (1);

[0042] Figure 2 DSC spectrum of Form A of the compound of formula (1);

[0043] Figure 3 TGA spectrum of Form A of the compound of formula (1);

[0044] Figure 4 XPRD pattern of Form B of the compound of formula (1);

[0045] Figure 5 DSC spectrum of Form B of the compound of formula (1);

[0046] Figure 6 TGA spectrum of Form B of the compound of formula (1);

[0047] Figure 7 XPRD pattern of Form C of the compound of formula (1);

[0048] Figure 8 DSC spectrum of Form C of the compound of formula (1);

[0049] Figure 9 TGA spectrum of Form C of the compound of formula (1);

[0050] Figure 10 XPRD pattern of Form D of the compound of formula (1);

[0051] Figure 11 DSC spectrum of Form D of the compound of formula (1);

[0052] Figure 12 TGA spectrum of Form D of the compound of formula (1). DETAILED DESCRIPTION

[0053] The present invention will be further described in detail below with reference to the examples, but is not limited thereto.

[0054] Test conditions of the instruments used in the experiment:

[0055] XRPD is an X-ray powder diffraction test: the measurement was carried out using a Bruker D8 X-ray diffractometer in accordance with the 2020 edition of the Chinese Pharmacopoeia, Part IV, General Chapter 0451, test conditions: Target: Cu; 40kV, 40mA.

[0056] DSC stands for differential scanning calorimetry: the measurement was carried out using a TA Q2000 differential scanning calorimeter in accordance with the general rules 0661 of Part IV of the 2020 edition of the Chinese Pharmacopoeia. The test conditions were: DSC: 30°C 10°C / min 300°C; TGA: 30°C 10°C / min 350°C.

[0057] Example 1 Properties of the Pharmaceutically Acceptable Salt of 2-{N-[4-(4-methyl)piperazine]phenyl}amino-5,6-dihydro-7-[3-(N-tert-butyl)sulfonamido]phenyl-pyrrolo[2,3-d]pyrimidine

[0058] 1. Melting point

[0059] Table 1

[0060] Salt type Melting point Decomposition temperature sulfate / 258.05℃ dihydrochloride / 271.44℃ Oxalate 155.10℃ 230℃ Methanesulfonate 164.22℃ 225℃ Citrate 135.39℃ 187.5℃

[0061] 2. Solubility

[0062] The test results show that compared with the free base, the solubility of its sulfate, dihydrochloride, oxalate, methanesulfonate and citrate in water is improved, especially the sulfate and dihydrochloride can significantly improve the solubility of the free base in water.

[0063] Table 2 Saturated solubility (in water) test of different salt types

[0064]

[0065] 3. Stability

[0066] The stability of the free base and five different types of pharmaceutically acceptable salts was investigated under two conditions.

[0067] To compare the solid-state stability of the five salt forms with the free base, the free base and its dihydrochloride, sulfate, methanesulfonate, oxalate, and citrate salts were subjected to stability evaluations at 30°C ± 2°C / 65% RH ± 5% RH and 40°C ± 2°C / 75% RH ± 5% RH for one month. The chemical stability of the stability test samples was assessed by HPLC for purity (HPLC method using area normalization).

[0068] Table 3: Summary of Stability Assessment

[0069]

[0070] Conclusion: No significant purity reduction was found for the dihydrochloride under the stability storage conditions of 30℃±2℃ / 65%RH±5%RH and 40℃±2℃ / 75%RH±5%RH, while the purity of the free base and its sulfate, methanesulfonate, oxalate and citrate all decreased to a certain extent under the same storage conditions.

[0071] 4. Hygroscopicity assessment

[0072] Referring to the guidelines for drug hygroscopicity testing in the 2020 edition of the Chinese Pharmacopoeia, hygroscopicity evaluations were conducted on the free base and its five salt forms. At a constant temperature of 25°C and a relative humidity of 80%, data on the changes in the mass of solid samples (free base and its dihydrochloride, sulfate, oxalate, methanesulfonate, and citrate) were collected after 24 hours of storage. The hygroscopicity evaluation is summarized in Table 4:

[0073] Table 4: Summary of Moisture Emission Evaluation

[0074] Salt type Weight gain after moisture induction Hygroscopicity sulfate 8.2％ Hygroscopic dihydrochloride 4.5％ Hygroscopic Oxalate 7.6％ Hygroscopic Methanesulfonate 7.8％ Hygroscopic Citrate 4.5％ Hygroscopic Free base 1.7％ Slightly hygroscopic

[0075] Conclusion: From the above hygroscopicity evaluation results, it can be seen that the free base is slightly hygroscopic, and the five salt forms are all hygroscopic, among which the dihydrochloride and citrate have relatively lower hygroscopicity, showing better physical and chemical properties.

[0076] Example 2 Preparation of 2-{N-[4-(4-methyl)piperazine]phenyl}amino-5,6-dihydro-7-[3-(N-tert-butyl)sulfonamido]phenyl-pyrrolo[2,3-d]pyrimidine dihydrochloride crystals

[0077] 1. Preparation of Form A

[0078] The compound of formula (1) (42 g) was added to a mixed solvent of 420 ml of anhydrous ethanol and 63 ml of purified water, heated and stirred to dissolve, stirred and crystallized, filtered, the filter cake was collected, and vacuum dried at 80°C to obtain the product (40 g, yield 95%).

[0079] X-ray powder diffraction showed that it was Form A, and its XRPD spectrum was as follows: Figure 1 The positions of its characteristic peaks are shown in Table 5 below. In the DSC spectrum, there is an endothermic peak at 273.68°C. Thermogravimetric analysis (TGA) shows a weight loss of 2.854% between 25°C and 100°C.

[0080] Table 5 Characteristic peaks of A crystal form

[0081]

[0082]

[0083] 2. Preparation of Form A

[0084] The compound of formula (1) (0.3 g) was added to 9 ml of anhydrous ethanol, stirred and slurried, stirred and crystallized, filtered, the filter cake was collected, and vacuum dried at 90°C to obtain the product (0.23 g, yield 77%), which was detected by XPRD to be Form A.

[0085] 3. Preparation of Form A

[0086] The compound of formula (1) (0.5 g) was added to 3 ml of anhydrous methanol, heated and stirred to dissolve, stirred and crystallized, filtered, the filter cake was collected, and vacuum dried at 60°C to obtain the product (0.21 g, yield 42%), which was detected by XPRD to be Form A.

[0087] 4. Preparation of Form B

[0088] The compound of formula (1) (3.66 g) was added to a mixed solvent of 73 ml of anhydrous ethanol and 4.5 ml of purified water, heated and stirred to dissolve, stirred and crystallized, filtered, the filter cake was collected, and dried in vacuo at 40°C to obtain the product (3.02 g, yield 82.5%).

[0089] X-ray powder diffraction showed that it was Form B, and its XRPD spectrum was as follows: Figure 4 The positions of its characteristic peaks are shown in Table 6 below. In the DSC spectrum, there is an endothermic peak at 266.04°C. Thermogravimetric analysis (TGA) shows a weight loss of 6.505% between 25°C and 100°C.

[0090] Table 6 Characteristic peaks of B crystal form

[0091]

[0092]

[0093] 5. Preparation of Form B

[0094] The compound of formula (1) (0.5 g) was added to 3 ml of anhydrous methanol, heated and stirred to dissolve, stirred and crystallized, filtered, the filter cake was collected, and vacuum dried at 30°C to obtain the product (0.26 g, yield 52%), which was detected by XPRD to be Form B.

[0095] 6. Preparation of Form B

[0096] The compound of formula (1) (0.3 g) was added to a mixed solvent of 9.5 ml of anhydrous methanol and 4 ml of methyl tert-butyl ether, heated and stirred to dissolve, stirred and crystallized, filtered, and the filter cake was collected and dried in vacuo at 25°C to obtain the product (0.21 g, yield 70%), which was detected by XPRD to be Form B.

[0097] 7. Preparation of Form C

[0098] The compound of formula (1) (0.3 g) was added to 9 ml of anhydrous ethanol, stirred and slurried, stirred and crystallized, filtered, the filter cake was collected, and dried in vacuo at 30°C to obtain the product (0.25 g, yield 83%).

[0099] X-ray powder diffraction showed that it was Form C, and its XRPD spectrum was as follows: Figure 7 The positions of its characteristic peaks are shown in Table 7 below. In the DSC spectrum, there is an endothermic peak at 274.4°C. Thermogravimetric analysis (TGA) shows a weight loss of 7.307% between 25°C and 100°C.

[0100] Table 7 Characteristic peaks of C crystal form

[0101]

[0102] 8. Preparation of Form C

[0103] The compound of formula (1) (0.3 g) was added to a mixed solvent of 7 ml of anhydrous methanol and 2.8 ml of ethyl acetate, heated and stirred to dissolve, stirred and crystallized, filtered, the filter cake was collected, and dried in vacuo at 40°C to obtain the product (0.26 g, yield 87%), which was detected by XPRD to be Form C.

[0104] 9. Preparation of Crystal Form D

[0105] The compound of formula (1) (0.3 g) was added to 4 ml of anhydrous methanol, stirred to dissolve, 16 ml of acetonitrile was added dropwise, stirred to crystallize, filtered, the filter cake was collected, and dried in vacuo at 30°C to obtain the product (0.17 g, yield 57%).

[0106] X-ray powder diffraction showed that it was Form D, and its XRPD spectrum was as follows: Figure 10 The positions of its characteristic peaks are shown in Table 8 below. In the DSC spectrum, there is an endothermic peak at 234.85°C. Thermogravimetric analysis (TGA) shows a weight loss of 6.25% between 25°C and 100°C.

[0107] Table 8 Characteristic peaks of D crystal form

[0108]

[0109]

[0110] Example 3 Stability Test of Crystal Form A, Crystal Form B, Crystal Form C, and Crystal Form D

[0111] Purity testing was carried out according to the 0512 method of the General Chapter 4 of the Chinese Pharmacopoeia 2015 edition.

[0112]

[0113]

[0114] Example 4A: Test of factors affecting the crystal form

[0115] Purity testing was carried out according to the 0512 method of the General Chapter 4 of the Chinese Pharmacopoeia 2015 edition.

[0116]

[0117]

[0118] Example 5 Solubility Determination of Crystal Form A, Crystal Form B, Crystal Form C, and Crystal Form D

[0119] According to the solubility test method of Part IV of the 2015 edition of the Chinese Pharmacopoeia, place this product in a certain volume of solvent at 25±2℃.

[0120] Shake vigorously for 30 seconds every 5 minutes and observe the dissolution within 30 minutes. The results are as follows:

[0121]

[0122] Example 6 Dissolution characteristics of the crystalline dihydrochloride form in the formulation

[0123] In vitro dissolution tests can objectively reflect the characteristics of the preparation and play an important guiding and predictive role in the dissolution and release of the drug in vivo. Using the same prescription and powder direct compression process, the finished product of the dihydrochloride crystal form A preparation can reach a dissolution rate of more than 85% within 15 minutes in pH 1.2 hydrochloric acid medium, pH 4.5 acetate buffer, pH 6.8 phosphate buffer (containing 0.05% SDS) and aqueous medium that simulate the in vivo environment, achieving a rapid release effect. However, due to the poor solubility of the raw material drug, the finished free base preparation can only reach a dissolution rate of more than 80% within 120 minutes in pH 1.2 hydrochloric acid medium and pH 4.5 acetate buffer. The dissolution effect in pH 6.8 phosphate buffer (containing 0.05% SDS) and aqueous medium is even worse, and the dissolution rate within 120 minutes does not exceed 20%.

[0124] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principles of the present invention should be included in the scope of protection of the present invention.

Claims

1. A crystalline form A of the compound of formula (1), 2-{N-[4-(4-methyl)piperazine]phenyl}amino-5,6-dihydro-7-[3-(N-tert-butyl)sulfonamido]phenyl-pyrrolo[2,3-d]pyrimidine dihydrochloride, characterized in that: Its X-ray powder diffraction pattern shows characteristic peaks at 2θ of 3.105, 5.989, 6.765, 8.478, 12.288, 13.315, 16.432, 16.985, 17.476, 19.370, 22.174, 24.166, 24.898, and 30.243;

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