Crystal polymorphism of JAK tyrosine kinase inhibitor and method for producing same

Abstract

The present invention relates to crystalline polymorphs of the compound 2-{3-[3-amino-4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]-1-(isopropylsulfonyl)azetidin-3-yl}acetonitrile (Formula A) and pharmaceutical compositions and uses thereof. TIFF2025506562000013.tif50130

Description

[Technical field]

[0001] The present application belongs to the chemical pharmaceutical field and relates to the crystal polymorphism of chemical drugs, in particular to the crystal polymorphism of the compound 2-{3-[3-amino-4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]-1-(isopropylsulfonyl)azetidin-3-yl}acetonitrile and a method for preparing the same. [Background technology]

[0002] 2-{3-[3-amino-4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]-1-(isopropylsulfonyl)azetidin-3-yl}acetonitrile (compound A) belongs to the JAK family of small molecule non-receptor tyrosine kinase inhibitors, whose JAK-STAT signaling pathway is closely related to inflammatory cytokines and tumors, and is widely involved in important biological processes such as cell proliferation, differentiation, metastasis and apoptosis, and the regulation of immune response and cell homeostasis in human health and disease processes. The structural formula of compound A is as follows: TIFF2025506562000002.tif50130

[0003] The synthesis method of this compound was disclosed in the invention patent CN201711248509.8, but the crystal form of the compound was not mentioned, and the crystal form of 2-{3-[3-amino-4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]-1-(isopropylsulfonyl)azetidin-3-yl}acetonitrile has not been reported in other literature. However, the crystal polymorphism of a drug has important significance to the physical properties, bioavailability, formulation quality and process of the drug, and when a drug with crystal polymorphism has a different crystal form, the difference in their physicochemical properties will affect the stability of the drug. When the drug is the same but has a different crystal form, their bioavailability may have a significant difference. Different crystal forms affect the dissolution rate of the drug. In addition, the difference in surface free energy of different crystal forms causes different binding forces between crystal particles, which affects the fluidity, particle uniformity, content uniformity and physical stability of the drug. Therefore, it is necessary to consider the crystal form. Summary of the Invention

[0004] The purpose of this application is to provide a polymorph of compound A, whose chemical name is 2-{3-[3-amino-4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]-1-(isopropylsulfonyl)azetidin-3-yl}acetonitrile, and a method for preparing the same. The synthesis method of compound A is disclosed in invention patent CN201711248509.8, and compound A prepared with reference to this invention patent is used to prepare the polymorph of this application.

[0005] According to one aspect, the present application provides a crystalline polymorph of Compound A.

[0006] The present application provides crystalline polymorphs of drug Compound A, which include multiple crystalline forms of Compound A, such as, for example, Form I, Form IIa, Form IIb, and Form III. The present application further provides crystalline forms of salts of Compound A.

[0007] The powder X-ray diffraction pattern of crystalline form I of 2-{3-[3-amino-4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]-1-(isopropylsulfonyl)azetidin-3-yl}acetonitrile of the present application is shown in FIG. 1, in which the measurement error of 2θ is ±0.2 degrees, and it includes several characteristic peaks between 0 and 40 degrees, as shown in the table below. TIFF2025506562000003.tif96165

[0008] The crystal form I according to the present application was shown to have no exothermic or endothermic peaks between 50 and 200°C in a differential scanning calorimeter (DSC), endothermic peaks at around 213.5°C, 219.6°C, and 229.0°C, and a clear exothermic peak between 213.5°C and 229.0°C. The differential scanning calorimeter curve of the crystal form I showed that this crystal form underwent a crystal transition during the process of heating and melting. The sample was shown to have endothermic and melted at about 213.5°C, and then to have exothermic and transitioned to a more stable crystal form, and when the temperature was further increased, the newly generated crystal form was shown to have endothermic and melted. The transition of the crystal form showed the relative magnitude of stability (melting point, lattice energy) between different crystal forms, but such a change occurred at a relatively high temperature (>200.0°C), which does not correspond to the stability of the crystal form under normal use conditions.

[0009] The crystalline form I according to the present application does not contain water of crystallization and does not contain a crystallization solvent.

[0010] The present application relates to crystalline form IIa of 2-{3-[3-amino-4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]-1-(isopropylsulfonyl)azetidin-3-yl}acetonitrile. The powder X-ray diffraction pattern of crystalline form IIa is shown in FIG. 3, in which the measurement error of 2θ is ±0.2 degrees, and several characteristic peaks included in the range of 0 to 40 degrees are shown in the following table. TIFF2025506562000004.tif99164

[0011] It was shown that the crystalline form IIa according to the present application had no endothermic peak between 50 and 200°C and had an endothermic peak around 220.2°C in differential scanning calorimetry (DSC).

[0012] The crystalline form IIa according to the present application does not contain water of crystallization and is anhydrous.

[0013] The present application relates to crystalline form IIb of 2-{3-[3-amino-4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]-1-(isopropylsulfonyl)azetidin-3-yl}acetonitrile. The powder X-ray diffraction pattern of crystalline form IIb is shown in FIG. 5, in which the measurement error of 2θ is ±0.2 degrees, and several characteristic peaks included in the range of 0 to 40 degrees are shown in the following table. TIFF2025506562000005.tif132162

[0014] The crystalline form IIb according to the present application was shown to have an endothermic peak between 50 and 200°C and at about 221.5°C in differential scanning calorimetry (DSC).

[0015] The crystalline form IIb according to the present application showed a weight loss of about 2.2% at 50 to 150°C in thermogravimetric analysis (TGA). The amount of weight loss was calculated to be about 0.5 water molecules based on the weight loss, and thus the crystalline form IIb was a 0.5 hydrate of compound A.

[0016] According to another aspect, the present application relates to an oral pharmaceutical composition comprising crystalline Form I of the present application.

[0017] In another aspect, the present application relates to an external pharmaceutical composition comprising the crystalline form IIb of the present application.

[0018] According to yet another aspect, the present application relates to the use of said crystalline form or said pharmaceutical composition in the manufacture of a medicament for treating a Janus kinase mediated disease.

[0019] According to yet another aspect, the present application relates to the use of said crystalline form or said pharmaceutical composition in the manufacture of a medicament for treating a Janus kinase mediated disease.

[0020] In another aspect, the present application relates to crystalline Form I or crystalline Form IIb for treating a Janus kinase mediated disease.

[0021] According to yet another aspect, the present application relates to a method of treating a Janus kinase mediated disease comprising administering to an individual in need thereof the above-mentioned crystalline form or pharmaceutical composition. [Brief description of the drawings]

[0022] [Figure 1] This is the X-ray powder diffraction (XRPD) pattern of crystalline form I. [Diagram 2] This is a differential scanning calorimeter (DSC) pattern of crystalline form I. [Diagram 3] This is the X-ray powder diffraction (XRPD) pattern of crystalline form IIa. [Figure 4] This is a differential scanning calorimeter (DSC) pattern of crystalline form IIa. [Diagram 5] This is a pattern obtained by single crystal X-ray diffraction (XRSD) measurement of crystalline form IIb. [Figure 6] This is the X-ray powder diffraction (XRPD) pattern of crystalline form IIb. [Figure 7] This is a differential scanning calorimeter (DSC) pattern of crystalline form IIb. [Figure 8] This is a thermogravimetric analysis (TGA) pattern of crystalline form IIb. [Figure 9] 1 shows the skin permeation rates of crystalline form I and crystalline form IIb in a skin permeation test. [Figure 10] This is the X-ray powder diffraction (XRPD) pattern of crystalline form III. [Figure 11] 1H-NMR pattern of crystalline form III after standing as a powder. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0023] Hereinafter, various embodiments of the present application will be described in more detail with reference to examples, so that those skilled in the art can better understand the present application and its advantages. Note that the following examples are provided to describe the technical solutions of the present application in detail without further limiting them, and should not be considered as limiting the scope of the present application, but are merely illustrative and representative of the present application.

[0024] Unless otherwise specified, the reagents used in these examples are chemically pure reagents, and their suppliers are Fuchen (Tianjin) Chemical Reagent Co., Ltd. The experimental equipment used in the examples includes a round-bottom flask, a Buchner funnel, a suction filter bottle, and an electromagnetic stirrer (DF-101S), all of which are common laboratory equipment, and the drying equipment is dried by a vacuum oven (DZF-6020 type) or a double cone (FZG-8 type).

[0025] Example 1 Preparation of Crystalline Form I of 2-{3-[3-amino-4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]-1-(isopropylsulfonyl)azetidin-3-yl}acetonitrile In a 250 mL flask, 20.0 g of the compound 2-{3-[3-amino-4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]-1-(isopropylsulfonyl)azetidin-3-yl}acetonitrile and 100 mL of N,N-dimethylformamide were added, stirred, dissolved, and insoluble matter was removed by filtration. 250 mL of purified water was added to the filtrate, stirred to crystallize, filtered, and the resulting solid was dried and then transferred to a 250 mL flask. 200 mL of anhydrous ethanol was then added, refluxed with heating for 3 hours, cooled, filtered, and the resulting solid was dried at 55 to 60 ° C until a constant weight was reached, giving 18.8 g of off-white solid (yield 94.0%, chemical purity 99.62%).

[0026] Example 2 Preparation of the phosphate salt of 2-{3-[3-amino-4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]-1-(isopropylsulfonyl)azetidin-3-yl}acetonitrile 2-{3-[3-amino-4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]-1-(isopropylsulfonyl)azetidin-3-yl}acetonitrile (400 mg, 1.0 mmol, 1.0 eq) was added to 20 mL of acetonitrile, and 85% phosphoric acid (350 mg, 3.0 mmol, 3.0 eq) was added at room temperature. The mixture was reacted overnight at room temperature, and then concentrated under reduced pressure to obtain an oily product that was not a salt.

[0027] Example 3 Preparation of Crystalline Form IIa of 2-{3-[3-amino-4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]-1-(isopropylsulfonyl)azetidin-3-yl}acetonitrile 5.0 g of the compound 2-{3-[3-amino-4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]-1-(isopropylsulfonyl)azetidin-3-yl}acetonitrile crystalline form I was added to a 100 mL flask, 100 mL of ethanol (95%) was added, and the mixture was refluxed with stirring for 5 h while increasing the temperature, cooled, and filtered to obtain a solid. The solid was dried in vacuum (0.1 MPa) at 80°C for 12 h to obtain 4.8 g of solid (yield 96.0%, purity by HPLC: 99.50%).

[0028] Example 4 Preparation of the phosphate salt of 2-{3-[3-amino-4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]-1-(isopropylsulfonyl)azetidin-3-yl}acetonitrile 2-{3-[3-amino-4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]-1-(isopropylsulfonyl)azetidin-3-yl}acetonitrile (400 mg, 1.0 mmol, 1.0 eq) was added to 20 mL of acetonitrile, and 85% phosphoric acid (350 mg, 3.0 mmol, 3.0 eq) was added at room temperature, and then the temperature was raised to 70° C. and the reaction was carried out for 3 hours. HPLC showed that the reaction system had become complicated (the area of ​​the main peak changed from 98% to 87%), and after concentration under reduced pressure, a non-salt oil was obtained.

[0029] Example 5 Preparation of 2-{3-[3-amino-4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]-1-(isopropylsulfonyl)azetidin-3-yl}acetonitrile hydrochloride 2-{3-[3-amino-4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]-1-(isopropylsulfonyl)azetidin-3-yl}acetonitrile (400 mg, 1.0 mmol, 1.0 eq) was added to 20 mL of acetonitrile, and 38% concentrated hydrochloric acid (290 mg, 3.0 mmol, 3.0 eq) was added at room temperature and allowed to react overnight at room temperature. HPLC showed that the impurities had increased, and after concentration under reduced pressure, an oil that was not a salt was obtained.

[0030] Example 6 Preparation of Crystalline Form IIb of 2-{3-[3-amino-4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]-1-(isopropylsulfonyl)azetidin-3-yl}acetonitrile In a 100 mL flask, 6.2 g of the compound 2-{3-[3-amino-4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]-1-(isopropylsulfonyl)azetidin-3-yl}acetonitrile was added, 25 mL of N,N-dimethylformamide was added, stirred and dissolved, and insoluble matter was removed by filtration. 50 mL of purified water was added to the filtrate, stirred to crystallize, and filtered. The obtained solid was dried at 55 to 60 ° C until a constant weight was reached, and 5.6 g of solid (yield 88.9%) was obtained.

[0031] Example 7 Preparation of 2-{3-[3-amino-4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]-1-(isopropylsulfonyl)azetidin-3-yl}acetonitrile hydrochloride 2-{3-[3-amino-4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]-1-(isopropylsulfonyl)azetidin-3-yl}acetonitrile (400 mg, 1.0 mmol, 1.0 eq) was added to 20 mL of acetonitrile, and 38% concentrated hydrochloric acid (290 mg, 3.0 mmol, 3.0 eq) was added at room temperature, and the mixture was then heated to 40° C. and reacted for 3 hours. HPLC showed that the amount of impurities in the reaction system had increased, and after concentration under reduced pressure, an oily product that was not a salt was obtained.

[0032] Example 8 Preparation of Crystalline Form III of 2-{3-[3-amino-4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]-1-(isopropylsulfonyl)azetidin-3-yl}acetonitrile 15 mg of 2-{3-[3-amino-4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]-1-(isopropylsulfonyl)azetidin-3-yl}acetonitrile was added to 3 mL of dioxane, then the mixture was heated to 40° C. to dissolve, clarified, filtered, and the solution was evaporated at 40° C. to crystallize, yielding crystalline form III. This crystalline form had poor crystallinity and stability, and after standing at room temperature, nuclear magnetic resonance analysis showed that about 10% impurities were formed.

[0033] TIFF2025506562000006.tif58143 TIFF2025506562000007.tif86130

[0034] Consideration of the stability of different crystal forms The crystal forms I, IIa, and IIb prepared in Examples 1, 3, and 6 were selected and the stability of the crystal forms I, IIa, and IIb was investigated according to the guiding principles for stability studies issued by the State Food and Drug Administration. The samples were packaged in polar pharmaceutical low-density polyethylene bags and the long-term stability was investigated for 24 months. The results are as follows: TIFF2025506562000008.tif82166 TIFF2025506562000009.tif83166 TIFF2025506562000010.tif82165

[0035] The above results of the study on the stability of different crystal forms show that the chemical stability of the compound 2-{3-[3-amino-4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]-1-(isopropylsulfonyl)azetidin-3-yl}acetonitrile crystal forms I and IIb is good. Crystal form IIa gradually absorbs water during the standing process and transforms into crystal form IIb, which causes the sample to absorb water and become heavier, resulting in a decrease in content.

[0036] Skin permeation test The crystal forms prepared in Examples 1 and 6 were selected to prepare gel formulations (the gel formulations may be prepared as follows: PEG400, propylene glycol, glycerin, ethanol, and the crystals of Examples 1 or 6 were uniformly stirred, then mixed with an EDTA aqueous solution, added to the swollen carbomer aqueous solution, and the pH was adjusted to 6 with a 10% triethanolamine solution to obtain a gel), and the skin permeation rates of each were measured by a skin permeation test (receptor medium 5.0% Tween 80, pH 5.5 phosphate). The test results are shown in Figures 9A and 9B, which show that the skin permeation performance of crystal form I is better.

[0037] Pharmacokinetics experiments The solid powder and its hydrogel formulation of Examples 1 and 6 were selected, and the pharmacokinetic data of rats administered orally in the stomach and by transdermal application were measured. SD male rats were divided into groups of 3 rats each, and the gel formulations of Examples 1 and 6 were administered once in the stomach, the gel formulation of Example 1 was administered intravenously, and the gel formulations of Examples 1 and 6 were administered by transdermal application. The animals were fasted overnight before the experiment, and the fasting period was from 10 hours before administration to 4 hours after administration. Blood samples were taken from the oral administration group and the transdermal administration group 0.25, 0.5, 1, 2, 4, 6, 8, and 24 hours after administration, and blood samples were taken from the intravenous administration group 0.083, 0.25, 0.5, 1, 2, 4, 6, 8, and 24 hours after administration. After anesthetizing with isoflurane using a small animal anesthesia machine, 0.3 mL of whole blood was collected from the orbital venous plexus and placed in a heparin anticoagulant tube. The sample was centrifuged at 4°C and 4000 rpm for 5 min, and the plasma was transferred to a centrifuge tube and stored at -80°C until analysis. The plasma sample was extracted by protein precipitation, and the extract was analyzed by LC / MS / MS. For the percutaneous administration group, 24 hours after percutaneous administration, the drug was washed off from the epidermis, and the skin at the administration site of the rat was peeled off, homogenized, and the drug content in the skin was analyzed by LC / MS / MS. The results of the pharmacokinetic experiment are shown in Tables 1 and 2. TIFF2025506562000011.tif74166 TIFF2025506562000012.tif37147

[0038] Form I and IIb are characterized by high crystallinity and good stability, and Form I has higher solubility and higher bioavailability when administered orally than Form IIb. Form IIb is less absorbent than Form I, but the drug concentration retained in the skin of animals is several times higher (e.g., 7 times higher) than Form I. Due to this unexpected feature, Form IIb is more suitable for cases where the drug is less likely to penetrate the system but is retained in the skin at a higher concentration.

Claims

1. Crystalline Form I of the compound 2-{3-[3-amino-4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]-1-(isopropylsulfonyl)azetidin-3-yl}acetonitrile, characterized in that the 2θ values ​​of characteristic peaks in a powder X-ray diffraction pattern are 6.48, 13.01, 15.38, 19.61, 20.68, 23.47, and 26.29, with a measurement error of ±0.2 degrees.

2. The crystalline form I according to claim 1, characterized in that the 2θ values ​​of characteristic peaks in the powder X-ray diffraction pattern of said crystalline form are 6.48, 7.68, 9.90, 13.01, 14.81, 15.38, 16.43, 16.87, 19.61, 20.68, 23.47, 26.29 and 32.23, with a measurement error of ±0.2 degrees.

3. Crystalline Form I according to claim 1, characterized in that the powder X-ray diffraction pattern of said crystal form is shown in Figure 1.

4. The crystalline form IIb of the compound 2-{3-[3-amino-4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]-1-(isopropylsulfonyl)azetidin-3-yl}acetonitrile is characterized in that the 2θ values ​​of characteristic peaks in a powder X-ray diffraction pattern are 7.93, 9.36, 11.29, 14.95, 20.96, 21.36, 21.77, 22.12, and 22.79, with a measurement error of ±0.2 degrees.

5. The crystalline form IIb according to claim 4, characterized in that the 2θ values ​​of characteristic peaks in the powder X-ray diffraction pattern of said crystalline form are 7.93, 9.36, 11.29, 14.69, 14.95, 15.13, 15.95, 19.93, 20.15, 20.96, 21.36, 21.77, 22.12, 22.79, 23.54, 24.14, 26.38, 28.74, and 29.15, with a measurement error of ±0.2 degrees.

6. The crystalline form IIb according to claim 4, characterized in that the powder X-ray diffraction pattern of said crystalline form is shown in Figure 6.

7. 7. Crystalline Form IIb according to any one of claims 4 to 6, wherein the crystalline form is a hemihydrate crystalline form of the compound 2-{3-[3-amino-4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]-1-(isopropylsulfonyl)azetidin-3-yl}acetonitrile.

8. 4. An oral pharmaceutical composition comprising crystalline form I according to any one of claims 1 to 3.

9. 8. An external pharmaceutical composition comprising the crystalline form IIb of any one of claims 4 to 7.

10. 10. Use of a crystalline form or pharmaceutical composition according to any one of claims 1 to 9 in the manufacture of a medicament for treating a Janus kinase mediated disease.

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