A polymorph of a JAK tyrosine kinase inhibitor and a preparation method thereof

Abstract

The present disclosure relates to 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), pharmaceutical compositions, and uses thereof. #imgabs0#

Description

Technical Field

[0001] The present application belongs to the field of chemical pharmaceuticals and relates to polymorphs of chemical drugs, and specifically to 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 preparation method thereof. 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) is a small molecule non-receptor tyrosine kinase inhibitor of the JAK class. 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, regulation of immune response, and cell homeostasis in human health and disease. The structural formula of Compound A is as follows:

[0003]

[0004] The synthesis method of this compound is disclosed in the invention patent CN201711248509.8, but the crystal form of the compound is not involved, and no other literature reports 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; and the polymorphic form of the drug is of great significance to the physical properties, bioavailability, quality and process of the drug. The differences in physicochemical properties between different crystal forms of polymorphic drugs affect the stability of the drug. The bioavailability of the same drug may vary significantly depending on the crystal form. Different crystal forms affect the dissolution rate of the drug. In addition, the difference in surface free energy of different crystal forms will cause different binding forces between crystalline particles, affecting the flowability, particle uniformity, content uniformity and physical stability of the drug. Therefore, it is necessary to study the crystal form. Summary of the Invention

[0005] The purpose of this application is to provide a polymorph of Compound A and a preparation method thereof. The chemical name of this compound is 2-{3-[3-amino-4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]-1-(isopropylsulfonyl)azetidin-3-yl}acetonitrile. The synthesis method of Compound A is disclosed in invention patent CN201711248509.8. Compound A is prepared with reference to this invention patent for the preparation of the polymorphs described in this application.

[0006] According to one aspect, the present application provides a polymorphic form of Compound A.

[0007] The present application provides polymorphs of the pharmaceutical compound A, including multiple crystalline forms of compound A, such as crystalline form I, crystalline form IIa, crystalline form IIb, and crystalline form III; crystalline forms of salts of compound A are also provided.

[0008] The 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, the powder X-ray diffraction pattern of the crystalline form I is shown in FIG1 , wherein the measurement error of 2θ is ±0.2 degrees, and there are multiple characteristic peaks between 0 and 40 degrees, as shown in the following table:

[0009] Table 1: X-ray diffraction data of Form I

[0010]

[0011] The crystal form I involved in the present application shows no exothermic and endothermic peaks between 50 and 200°C in differential scanning calorimetry (DSC), and has endothermic peaks near 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 calorimetry curve of crystal form I shows that the crystal form will undergo crystal transformation during the melting process of heating. It is manifested in that after the sample melts at around 213.5°C, it releases heat and transforms into a crystal form with higher stability. When the temperature is further increased, the newly formed crystal form undergoes endothermic melting. The transformation of the crystal form shows the relative size of the stability (melting point, lattice energy) between different crystal forms, but this change occurs at a higher temperature (>200.0°C) and does not correspond to the stability of the crystal form under conventional conditions of use.

[0012] The crystalline form I involved in the present application does not contain crystal water and does not contain a crystallization solvent.

[0013] The present application relates to 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 Form IIa is shown in FIG3 , wherein the measurement error of 2θ is ±0.2 degrees, and multiple characteristic peaks between 0 and 40 degrees are shown in the following table:

[0014] Table 2: X-ray diffraction data of Form IIa

[0015]

[0016] The crystal form IIa involved in the present application shows no endothermic peak between 50 and 200°C in differential scanning calorimetry (DSC), but has an endothermic peak near 220.2°C.

[0017] The crystalline form IIa involved in the present application does not contain crystalline water and is an anhydrate.

[0018] The present application relates to 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 Form IIb is shown in FIG5 , wherein the measurement error of 2θ is ±0.2 degrees, and multiple characteristic peaks between 0 and 40 degrees are shown in the following table:

[0019] Table 3: X-ray diffraction data of Form IIb

[0020]

[0021]

[0022] The crystal form IIb involved in the present application shows an endothermic peak between 50 and 200°C in differential scanning calorimetry (DSC), and an endothermic peak near 221.5°C.

[0023] The crystal form IIb involved in the present application shows a weight loss of about 2.2% at 50-150°C in thermogravimetric analysis (TGA). According to the weight loss calculation, the weight loss is about 0.5 molecules of water. Therefore, the crystal form IIb is a 0.5 molecule hydrate of compound A.

[0024] According to another aspect, the present application relates to an oral pharmaceutical composition comprising the crystalline form I of the present application.

[0025] According to another aspect, the present application relates to a pharmaceutical composition for external use, comprising the crystal form IIb of the present application.

[0026] According to yet another aspect, the present application relates to use of the above-mentioned crystal form or the above-mentioned pharmaceutical composition in the preparation of a drug for treating a disease mediated by Janus kinase.

[0027] According to another aspect, the present application relates to use of the above-mentioned crystal form or the above-mentioned pharmaceutical composition in the preparation of a drug for treating a disease mediated by Janus kinase.

[0028] According to another aspect, the present application relates to Form I or Form IIb for use in treating a disease mediated by Janus kinase.

[0029] According to yet another aspect, the present application relates to a method for treating a disease mediated by Janus kinase, comprising administering the above-mentioned crystalline form or pharmaceutical composition to an individual in need thereof. BRIEF DESCRIPTION OF THE DRAWINGS

[0030] Figure 1: X-ray powder diffraction (XRPD) spectrum of Form I.

[0031] Figure 2: Differential scanning calorimetry (DSC) spectrum of Form I.

[0032] Figure 3: X-ray powder diffraction (XRPD) spectrum of Form IIa.

[0033] Figure 4: Differential scanning calorimetry (DSC) spectrum of Form IIa.

[0034] Figure 5: Single crystal X-ray diffraction (XRSD) test spectrum of Form IIb.

[0035] Figure 6: X-ray powder diffraction (XRPD) spectrum of Form IIb.

[0036] Figure 7: Differential scanning calorimetry (DSC) spectrum of Form IIb.

[0037] Figure 8: Thermogravimetric analysis (TGA) spectrum of Form IIb.

[0038] Figure 9: Transdermal penetration rates of Form I and Form IIb in transdermal testing .

[0039] Figure 10 X-ray powder diffraction (XRPD) spectrum of Form III.

[0040] Figure 11: Form III powder after standing 1 H-NMR spectrum. DETAILED DESCRIPTION

[0041] Hereinafter, various embodiments of the present application will be described in more detail with reference to the examples, so that those skilled in the art can better understand the present application and its advantages. It should be noted that the following examples further illustrate the technical solutions of the present application in non-limiting detail and should not be considered as limiting the scope of the present application, but are merely exemplary and representative of the present application.

[0042] Unless otherwise specified, all reagents used in these examples were chemically pure and were supplied by Fuchen (Tianjin) Chemical Reagent Co., Ltd. The experimental instruments used in the examples, including round-bottom flasks, Büchner funnels, suction flasks, and magnetic stirring apparatus (DF-101S), were all general laboratory instruments. Drying was performed using a vacuum oven (DZF-6020) or a double cone oven (FZG-8).

[0043] Example 1

[0044] 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 Crystal Form I

[0045] 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 to dissolve, filtered, and the insoluble matter was removed. 250 mL of purified water was added to the filtrate, stirred for crystallization, and filtered. The resulting solid was dried and transferred to a 250 mL flask. 200 mL of anhydrous ethanol was added, the temperature was raised to reflux for 3 h, cooled, and filtered. The resulting solid was dried at 55-60 ° C to constant weight to give 18.8 g of an off-white solid with a yield of 94.0% and a chemical purity of 99.62%.

[0046] Example 2

[0047] 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 phosphate

[0048] -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 reaction was allowed to proceed overnight at room temperature and concentrated under reduced pressure to an oily product without forming a salt.

[0049] Example 3

[0050] 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 Form IIa

[0051] In a 100 mL flask, 5.0 g of type I crystals of compound 2-{3-[3-amino-4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]-1-(isopropylsulfonyl)azetidin-3-yl}acetonitrile were added, and 100 mL of ethanol (95%) was added. The mixture was heated and stirred at reflux for 5 h. The temperature was lowered and the mixture was filtered to obtain a solid. The obtained solid was dried at 80°C under vacuum (0.1 MPa) for 12 h to obtain 4.8 g of a solid with a yield of 96.0% and a purity of 99.50% as determined by HPLC.

[0052] Example 4

[0053] 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 phosphate

[0054] 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. 85% phosphoric acid (350 mg, 3.0 mmol, 3.0 eq) was added at room temperature. After addition, the temperature was raised to 70°C and the reaction was allowed to react for 3 hours. HPLC analysis showed that the reaction system had become complex (the main peak area changed from 98% to 87%). After concentration under reduced pressure, an oil was obtained, and no salt was formed.

[0055] Example 5

[0056] 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

[0057] 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. The reaction was allowed to proceed overnight at room temperature. HPLC showed an increase in impurities. After concentration under reduced pressure, the mixture was an oily substance and no salt was formed.

[0058] Example 6

[0059] 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 Form IIb

[0060] 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 and 25 mL of N,N-dimethylformamide were added. The mixture was stirred to dissolve and filtered to remove insoluble matter. 50 mL of purified water was added to the filtrate, stirred for crystallization, filtered, and the resulting solid was dried at 55-60°C to constant weight to obtain 5.6 g of solid with a yield of 88.9%.

[0061] Example 7

[0062] 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

[0063] 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. 38% concentrated hydrochloric acid (290 mg, 3.0 mmol, 3.0 eq) was added at room temperature. After addition, the mixture was heated to 40°C and reacted for 3 hours. HPLC analysis showed an increase in impurities in the reaction system. After concentration under reduced pressure, the product was an oil, with no salt formed.

[0064] Example 8

[0065] 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 Form III

[0066] 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, and the temperature was raised to 40°C to dissolve and clarify. After filtration, the solution was evaporated at 40°C to crystallize to obtain Form III. The crystallinity and stability of this form are poor. After stability is allowed to stand, nuclear magnetic resonance shows that about 10% of impurities are generated.

[0067] Investigation of the solubility of different crystal forms

[0068]

[0069]

[0070] Stability investigation of different crystal forms

[0071] The crystal samples prepared in Examples 1, 3, and 6 were selected and the stability of the prepared crystal forms I, IIa, and IIb was investigated in accordance with the stability study guidelines issued by the State Food and Drug Administration. The samples were packaged in pharmaceutical low-density polyethylene bags and the long-term stability was investigated for 24 months. The results are as follows:

[0072] Long-term 24-month stability of Compound A Form I

[0073]

[0074] Long-term 24-month stability of Compound A Form IIa

[0075]

[0076] Long-term 24-month stability of Compound A Form IIb

[0077]

[0078] The stability tests of the various crystalline forms above demonstrate that Forms I and 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 exhibit excellent chemical stability. Form IIa gradually absorbs water over time, transforming into Form IIb. This water absorption increases the sample's weight, leading to a decrease in sample content.

[0079] Transdermal test

[0080] The crystal forms prepared in Examples 1 and 6 were selected and prepared into gel formulations, respectively. (The gel formulations can be prepared by uniformly stirring PEG400, propylene glycol, glycerol, ethanol, and the crystals of Example 1 or 6, followed by mixing with an EDTA aqueous solution, adding the mixture to a swollen carbomer aqueous solution, and adjusting the pH to 6 with a 10% triethanolamine solution to obtain a gel.) The transdermal penetration rates of the crystal forms were determined by transdermal testing (receiving medium: 5.0% Tween 80, pH 5.5 phosphate). The test results are shown in Figures 9A and 9B, which indicate that Form I has better transdermal performance.

[0081] Pharmacokinetic experiments

[0082] The solid powders of Examples 1 and 6 and their hydrogel preparations were selected to measure the PK of rats administered orally by gavage and transdermal application. Male SD rats were divided into groups of 3 per group and administered orally by single gavage of Examples 1 and 6, intravenously by Example 1, and transdermally by gel preparations of Examples 1 and 6. The animals were fasted overnight before the experiment, and the fasting period was from 10 hours before administration to 4 hours after administration. Blood was collected from the oral group and transdermal group at 0.25, 0.5, 1, 2, 4, 6, 8 and 24 hours after administration, and from the intravenous group at 0.083, 0.25, 0.5, 1, 2, 4, 6, 8 and 24 hours after administration. After anesthesia with isoflurane using a small animal anesthesia machine, 0.3 mL of whole blood was collected from the fundus venous plexus and placed in a heparin anticoagulant tube. The sample was centrifuged at 4°C and 4000 rpm for 5 minutes. The plasma was transferred to a centrifuge tube and stored at -80°C until analysis. Plasma samples were extracted using protein precipitation, and the extracts were analyzed by LC / MS / MS. In the skin administration group, after 24 hours of transdermal administration and after washing away the epidermal drug, the skin at the administration site of the rats was peeled and homogenized, followed by LC / MS / MS analysis of drug content. The pharmacokinetic results are shown in Tables 1 and 2.

[0083] Table 1. Pharmacokinetic parameters of male rats after administration of the compounds of the examples

[0084]

[0085] Table 2. Drug concentrations in the skin 24 hours after transdermal administration

[0086]

[0087] Form I and Form IIb are characterized by high crystallinity and good stability. Form I has higher solubility and higher oral bioavailability than Form IIb. Although Form IIb has inferior transdermal absorption to Form I, the drug concentration retained in the skin of its topical preparations is significantly higher than that of Form I by several times (e.g., up to 7 times or more). This unexpected feature makes it more suitable for use in situations where systemic penetration is low and the drug retention concentration in the skin is higher.

Claims

1. 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 the characteristic peaks in the powder X-ray diffraction pattern of the crystalline form I are 6.48, 13.01, 15.38, 19.61, 20.68, 23.47, and 26.29, and the measurement error is ±0.2 degrees.

2. The crystalline form I according to claim 1, characterized in that The 2θ values of the characteristic peaks in the powder X-ray diffraction pattern of the crystal 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, and the measurement error is ±0.2 degrees.

3. The crystalline form I according to claim 1, characterized in that The powder X-ray diffraction pattern of the crystal form is shown in FIG1 .

4. 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, characterized in that: The crystalline form is a crystalline form of a hemihydrate 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 the 2θ values of the characteristic peaks in its 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, and the measurement error is ±0.2 degrees.

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

6. The crystalline form IIb according to claim 4, characterized in that The powder X-ray diffraction pattern of the crystal form is shown in FIG6 .

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

8. A pharmaceutical composition for external use, comprising the crystalline form IIb according to any one of claims 4 to 6.

9. Use of the crystalline form or pharmaceutical composition according to any one of claims 1 to 8 in the preparation of a medicament for treating a disease mediated by Janus kinase.

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