Crystalline polymorphs of JAK tyrosine kinase inhibitors and methods for producing the 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] This application belongs to the field of chemical pharmaceuticals and relates to crystal polymorphs of chemical drugs. Specifically, it relates to the crystal 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 and a method for producing the same.

Background Art

[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 non-receptor tyrosine kinase inhibitors of JAK small molecules. Its 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, apoptosis, immune response, and regulation of cell homeostasis in the process of human health and disease. The structural formula of Compound A is as follows. TIFF0007857417000001.tif50130

[0003] The synthesis method for this compound is disclosed in the invention's patent CN201711248509.8, but the crystalline form of the compound is not mentioned, and the crystalline form of 2-{3-[3-amino-4-(7H-pyrrolo[2,3-d]pyrimidine-4-yl)-1H-pyrazole-1-yl]-1-(isopropylsulfonyl)azetidine-3-yl}acetonitrile has not been reported in other literature. However, the crystalline polymorphism of a drug is of significant importance to its physical properties, bioavailability, formulation quality, and process. Drugs with crystalline polymorphism exhibit differences in physicochemical properties depending on the crystalline form, which affects the drug's stability. When the drug is the same but has different crystalline forms, their bioavailability may differ significantly. Different crystalline forms affect the drug's dissolution rate. Furthermore, differences in the surface free energy of different crystalline forms result in different bonding forces between crystal particles, affecting the drug's fluidity, particle uniformity, content uniformity, and physical stability. Therefore, the crystalline form needs to be considered. [Overview of the project]

[0004] The object of this application is to provide a crystalline polymorph of compound A, whose chemical name is 2-{3-[3-amino-4-(7H-pyrrolo[2,3-d]pyrimidine-4-yl)-1H-pyrazole-1-yl]-1-(isopropylsulfonyl)azetidine-3-yl}acetonitrile, and a method for producing the same. The method for synthesizing compound A is disclosed in the invention patent CN201711248509.8, and compound A produced by reference to said invention patent is used in the production of the crystalline polymorph in this application.

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

[0006] This application provides crystalline polymorphs of drug compound A, which include many crystalline forms of compound A, such as crystalline form I, crystalline form IIa, crystalline form IIb, and crystalline form III. This application further provides crystalline forms of salts of compound A.

[0007] Figure 1 shows the powder X-ray diffraction pattern of crystalline form I of 2-{3-[3-amino-4-(7H-pyrrolo[2,3-d]pyrimidine-4-yl)-1H-pyrazole-1-yl]-1-(isopropylsulfonyl)azetidine-3-yl}acetonitrile of the present application. The measurement error of 2θ is ±0.2 degrees, and as shown in the table below, it contains multiple characteristic peaks between 0 and 40 degrees. TIFF0007857417000002.tif96165

[0008] Crystal type I according to this application was shown to have no exothermic or endothermic peaks between 50 and 200°C in differential scanning calorimeter (DSC) measurements, but had endothermic peaks around 213.5°C, 219.6°C, and 229.0°C, with a clear exothermic peak between 213.5°C and 229.0°C. The differential scanning calorimetry curve of crystal type I showed that a crystal transition occurred during the heating and melting process. It was shown that after the sample melted endothermically at around 213.5°C, it exothermically transitioned to a more stable crystal type, and when the temperature rose further, the newly formed crystal type melted endothermically. The crystal transition showed the relative magnitude of stability (melting point, lattice energy) between different crystal types, but such changes occurred at relatively high temperatures (>200.0°C) and do not correspond to the stability of the crystal type under normal operating conditions.

[0009] Crystal type I relating to this application does not contain crystal water and does not contain a crystal solvent.

[0010] This application relates to crystalline form IIa of 2-{3-[3-amino-4-(7H-pyrrolo[2,3-d]pyrimidine-4-yl)-1H-pyrazole-1-yl]-1-(isopropylsulfonyl)azetidine-3-yl}acetonitrile. The powder X-ray diffraction pattern of crystalline form IIa is shown in Figure 3, where the measurement error of 2θ is ±0.2 degrees, and several characteristic peaks contained between 0 and 40 degrees are shown in the table below. TIFF0007857417000003.tif99164

[0011] Crystal type IIa according to the present application was shown to have no endothermic peaks between 50 and 200°C, but to have an endothermic peak around 220.2°C, as measured by differential scanning calorimeter (DSC).

[0012] Crystal type IIa relating to this application does not contain crystal water and is anhydrous.

[0013] This application relates to crystalline form IIb of 2-{3-[3-amino-4-(7H-pyrrolo[2,3-d]pyrimidine-4-yl)-1H-pyrazole-1-yl]-1-(isopropylsulfonyl)azetidine-3-yl}acetonitrile. The powder X-ray diffraction pattern of crystalline form IIb is shown in Figure 5, where the measurement error of 2θ is ±0.2 degrees, and several characteristic peaks contained between 0 and 40 degrees are shown in the table below. TIFF0007857417000004.tif132162

[0014] Crystal type IIb according to this application was shown to have an endothermic peak between 50 and 200°C, and an endothermic peak around 221.5°C, as measured by differential scanning calorimeter (DSC).

[0015] In thermogravimetric analysis (TGA), the crystalline form IIb of the present application showed a weight loss of approximately 2.2% at 50-150°C. Based on this weight loss, the amount of weight loss was calculated to be approximately 0.5 molecules of water, so the crystalline form IIb was the 0.5-hydrate of compound A.

[0016] In another embodiment, the present application relates to an oral pharmaceutical composition comprising crystalline form I of the present application.

[0017] In another embodiment, the present application relates to a topical pharmaceutical composition comprising the crystalline form IIb of the present application.

[0018] In another embodiment, the present application relates to the use of the above-mentioned crystalline form or pharmaceutical composition in the manufacture of a drug for treating Janus kinase-mediated diseases.

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

[0020] According to another aspect, the present application relates to crystalline form I or crystalline form IIb for treating Janus kinase-mediated diseases.

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

Brief Description of the Drawings

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

Modes for Carrying Out the Invention

[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 may better understand the present application and its advantages. The following examples are intended to illustrate the technical concept of the present application in detail without further limiting it, and should not be considered to restrict the scope of the present application; they are merely illustrative descriptions and typical representatives of the present application.

[0024] Unless otherwise specified, the reagents used in these examples are chemically pure, and their supplier is Fuchen (Tianjin) Chemical Reagents Co., Ltd. The laboratory equipment used in the examples includes round-bottom flasks, Buchner funnels, suction filtration bottles, and electromagnetic stirrers (DF-101S), all of which are common laboratory equipment. Drying is performed using a vacuum oven (DZF-6020) or a double cone oven (FZG-8).

[0025] Example 1 Preparation of crystalline form I of 2-{3-[3-amino-4-(7H-pyrrolo[2,3-d]pyrimidine-4-yl)-1H-pyrazole-1-yl]-1-(isopropylsulfonyl)azetidine-3-yl}acetonitrile 20.0 g of the compound 2-{3-[3-amino-4-(7H-pyrrolo[2,3-d]pyrimidine-4-yl)-1H-pyrazole-1-yl]-1-(isopropylsulfonyl)azetidine-3-yl}acetonitrile and 100 mL of N,N-dimethylformamide were added to a 250 mL flask, stirred to dissolve, 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 transferred to a 250 mL flask. Subsequently, 200 mL of anhydrous ethanol was added, and the mixture was refluxed while increasing the temperature for 3 hours, cooled, filtered, and the resulting solid was dried at 55-60°C until a constant weight was obtained, yielding 18.8 g of off-white solid (yield 94.0%, chemical purity 99.62%).

[0026] Example 2 Preparation of phosphate of 2-{3-[3-amino-4-(7H-pyrrolo[2,3-d]pyrimidine-4-yl)-1H-pyrazole-1-yl]-1-(isopropylsulfonyl)azetidine-3-yl}acetonitrile 2-{3-[3-amino-4-(7H-pyrrolo[2,3-d]pyrimidine-4-yl)-1H-pyrazole-1-yl]-1-(isopropylsulfonyl)azetidine-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 after concentration under reduced pressure, an oily substance, not a salt, was obtained.

[0027] Example 3 Preparation of crystalline form IIa of 2-{3-[3-amino-4-(7H-pyrrolo[2,3-d]pyrimidine-4-yl)-1H-pyrazole-1-yl]-1-(isopropylsulfonyl)azetidine-3-yl}acetonitrile 5.0 g of crystalline form I of the compound 2-{3-[3-amino-4-(7H-pyrrolo[2,3-d]pyrimidine-4-yl)-1H-pyrazole-1-yl]-1-(isopropylsulfonyl)azetidine-3-yl}acetonitrile was added to a 100 mL flask, 100 mL of ethanol (95%) was added, the mixture was stirred under increasing heat for 5 hours under reflux, cooled, filtered, and a solid was obtained. The obtained solid was dried under vacuum (0.1 MPa) at 80°C for 12 hours to obtain 4.8 g of solid (yield 96.0%, purity by HPLC: 99.50%).

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

[0029] Example 5 Preparation of 2-{3-[3-amino-4-(7H-pyrrolo[2,3-d]pyrimidine-4-yl)-1H-pyrazole-1-yl]-1-(isopropylsulfonyl)azetidine-3-yl}acetonitrile hydrochloride 2-{3-[3-amino-4-(7H-pyrrolo[2,3-d]pyrimidine-4-yl)-1H-pyrazole-1-yl]-1-(isopropylsulfonyl)azetidine-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. The mixture was reacted overnight at room temperature. HPLC showed an increase in impurities, and after concentration under reduced pressure, a non-salt oily substance was obtained.

[0030] Example 6 Preparation of crystalline form IIb of 2-{3-[3-amino-4-(7H-pyrrolo[2,3-d]pyrimidine-4-yl)-1H-pyrazole-1-yl]-1-(isopropylsulfonyl)azetidine-3-yl}acetonitrile 6.2 g of the compound 2-{3-[3-amino-4-(7H-pyrrolo[2,3-d]pyrimidine-4-yl)-1H-pyrazole-1-yl]-1-(isopropylsulfonyl)azetidine-3-yl}acetonitrile was added to a 100 mL flask, 25 mL of N,N-dimethylformamide was added, and the mixture was stirred to dissolve it. Insoluble matter was removed by filtration, 50 mL of purified water was added to the filtrate, and the mixture was stirred to crystallize it. The mixture was filtered, and the resulting solid was dried at 55-60°C until a constant weight was obtained, yielding 5.6 g of solid (yield 88.9%).

[0031] Example 7 Preparation of 2-{3-[3-amino-4-(7H-pyrrolo[2,3-d]pyrimidine-4-yl)-1H-pyrazole-1-yl]-1-(isopropylsulfonyl)azetidine-3-yl}acetonitrile hydrochloride 2-{3-[3-amino-4-(7H-pyrrolo[2,3-d]pyrimidine-4-yl)-1H-pyrazole-1-yl]-1-(isopropylsulfonyl)azetidine-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. The mixture was then heated to 40°C and reacted for 3 hours. HPLC showed an increase in impurities in the reaction system, and after concentration under reduced pressure, an oily substance (not a salt) was obtained.

[0032] Example 8 Preparation of crystalline form III of 2-{3-[3-amino-4-(7H-pyrrolo[2,3-d]pyrimidine-4-yl)-1H-pyrazole-1-yl]-1-(isopropylsulfonyl)azetidine-3-yl}acetonitrile 15 mg of 2-{3-[3-amino-4-(7H-pyrrolo[2,3-d]pyrimidine-4-yl)-1H-pyrazole-1-yl]-1-(isopropylsulfonyl)azetidine-3-yl}acetonitrile was added to 3 mL of dioxane, then heated to 40°C to dissolve, clarified, filtered, and the solution was evaporated at 40°C to crystallize and obtain crystal form III. This crystal form had poor crystallinity and stability, and after stable standing, nuclear magnetic resonance analysis showed that approximately 10% impurities had formed.

[0033] TIFF0007857417000005.tif58143 TIFF0007857417000006.tif86130

[0034] Consideration of the stability of different crystal forms Samples of the crystalline forms produced in Examples 1, 3, and 6 were selected, and the stability of the produced crystalline forms I, IIa, and IIb was examined in accordance with the guiding principles for stability studies issued by the State Food and Drug Administration. The samples were packaged in polarized low-density polyethylene bags for pharmaceutical use, and the long-term stability for 24 months was examined. The results of the examination are as follows. TIFF0007857417000007.tif82166 TIFF0007857417000008.tif83166 TIFF0007857417000009.tif82165

[0035] The results of the analysis of the stability of the different crystal forms described above showed that crystal forms I and IIb of the compound 2-{3-[3-amino-4-(7H-pyrrolo[2,3-d]pyrimidine-4-yl)-1H-pyrazole-1-yl]-1-(isopropylsulfonyl)azetidine-3-yl}acetonitrile have good chemical stability. Crystal form IIa gradually absorbed water during the standing process, converting to crystal form IIb, causing the sample to become heavier due to water absorption and resulting in a decrease in content.

[0036] Skin permeability test A gel formulation was prepared using the crystalline forms produced in Examples 1 and 6 (the gel formulation may be prepared as follows: PEG400, propylene glycol, glycerin, ethanol, and the crystalline form from Example 1 or 6 were uniformly stirred, then mixed with an EDTA aqueous solution, added to a swollen carbomer aqueous solution, and the pH was adjusted to 6 with a 10% triethanolamine solution to obtain a gel). The skin penetration rate of each (receptor medium 5.0% Tween 80, pH 5.5 phosphate) was measured by a skin penetration test. The test results are shown in Figures 9A and 9B, demonstrating that crystalline form I exhibited better skin penetration performance.

[0037] Pharmacokinetic experiments The solid powders and hydrogel formulations of Examples 1 and 6 were selected, and pharmacokinetic data were measured in rats administered orally via gastric injection and transdermally. Three male SD rats were divided into each group. Each group received a single intragastric administration of the gel formulations of Examples 1 and 6, intravenously administered the gel formulation of Example 1, and transdermally administered the gel formulations of Examples 1 and 6. The animals were fasted overnight before the experiment, with the fasting period ranging from 10 hours before administration to 4 hours after administration. Blood samples were collected from the oral and transdermal administration groups at 0.25, 0.5, 1, 2, 4, 6, 8, and 24 hours after administration, while blood samples were collected from the intravenous administration group at 0.083, 0.25, 0.5, 1, 2, 4, 6, 8, and 24 hours after administration. After anesthetizing rats 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 minutes, and the plasma was transferred to a centrifuge tube and stored at -80°C until analysis. The sample in the plasma was extracted by protein precipitation, and the extract was analyzed by LC / MS / MS. In the transdermal administration group, 24 hours after transdermal administration, the drug was washed off the epidermis, the skin at the administration site was peeled off, homogenized, and the drug content in the skin was analyzed by LC / MS / MS. The results of the pharmacokinetic experiments are shown in Tables 1 and 2. TIFF0007857417000010.tif74166 TIFF0007857417000011.tif37147

[0038] Crystalline forms I and IIb are characterized by high crystallinity and good stability. Crystalline form I has higher solubility than crystalline form IIb and exhibits higher bioavailability when administered orally. Crystalline form IIb has inferior transdermal absorption compared to crystalline form I, but the concentration of the drug retained on the skin of animals when applied topically is significantly several times higher (e.g., more than 7 times higher) than that of crystalline form I. This unexpected characteristic makes it more suitable for situations where penetration into the system is low but drug retention in the skin is high.

Claims

1. A crystal of crystalline form I of 2-{3-[3-amino-4-(7H-pyrrolo[2,3-d]pyrimidine-4-yl)-1H-pyrazole-1-yl]-1-(isopropylsulfonyl)azetidine-3-yl}acetonitrile, A crystal characterized by having 2θ values ​​of characteristic peaks in its powder X-ray diffraction pattern of 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 crystal according to Claim 1, A crystal characterized in that the 2θ values ​​of the characteristic peaks in the powder X-ray diffraction pattern 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, and the measurement error is ±0.2 degrees.

3. A crystal of crystalline form IIb of 2-{3-[3-amino-4-(7H-pyrrolo[2,3-d]pyrimidine-4-yl)-1H-pyrazole-1-yl]-1-(isopropylsulfonyl)azetidine-3-yl}acetonitrile, A crystal characterized by having 2θ values ​​of characteristic peaks in its powder X-ray diffraction pattern of 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.

4. The crystal according to claim 3, A crystal characterized in that the 2θ values ​​of characteristic peaks in the powder X-ray diffraction pattern 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, and the measurement error is ±0.2 degrees.

5. The crystal according to claim 3, A crystal characterized by being a hemihydrate.

6. The crystal according to claim 4, A crystal characterized by being a hemihydrate.

7. An oral pharmaceutical composition comprising the crystal described in claim 1 or 2.

8. A topical pharmaceutical composition comprising the crystal described in any one of claims 3 to 6.

9. Use of the crystal according to any one of claims 1 to 6 in the manufacture of a drug for treating Janus kinase-mediated diseases.

10. Use of the oral pharmaceutical composition according to claim 7 in the manufacture of a drug for treating Janus kinase-mediated diseases.

11. Use of the topical pharmaceutical composition according to claim 8 in the manufacture of a drug for treating Janus kinase-mediated diseases.

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