Crystal form of JAK kinase inhibitor

Abstract

The invention relates to a crystal form of a JAK kinase inhibitor, in particular to a polymorphic form of 2-{3-[3-amino-4-(7H-pyrrolo [2, 3-d] pyrimidine-4-yl)-1H-pyrazol-1-yl]-1-(isopropylsulfonyl) azetidin-3-yl} acetonitrile and a preparation method of the polymorphic form of the 2-{3-[3-amino-4-(7H-pyrrolo [2, 3-d] pyrimidine-4-yl)-1H-pyrazol-1-yl]-1-(isopropylsulfonyl) azetidin-3-yl} acetonitrile. The present application also relates to pharmaceutical compositions containing at least one polymorph and therapeutic or prophylactic uses of the polymorph and compositions.

Description

Technical Field

[0001] The present application relates to a polymorph 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 a preparation method thereof, as well as a pharmaceutical composition and an ointment dosage form containing at least one polymorph, and the therapeutic or preventive use of the polymorph and the composition. 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 the JAK class of small molecule non-receptor tyrosine kinase inhibitors, and 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 field needs to further study the crystal form of the compound A, which needs to have a crystalline or amorphous form with physical properties suitable for reliable formulation and manufacturing, including filterability, hygroscopicity, fluidity, solubility, etc. Summary of the invention

[0003] The present application provides a polymorph 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, a preparation method and an application thereof.

[0004] In the first aspect, the present application relates to a 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, wherein the crystalline form has an X-ray powder diffraction pattern including peaks at diffraction angles 2θ of 8.2°±0.2°, 14.5°±0.2° and 16.4°±0.2°.

[0005] In one embodiment, the X-ray powder diffraction pattern of the crystalline form further includes at least one peak located at diffraction angles 2θ of 10.6°±0.2°, 15.0°±0.2°, and 22.2°±0.2°.

[0006] In one embodiment, the X-ray powder diffraction pattern of the crystalline form further includes at least one peak located at diffraction angles 2θ of 13.6°±0.2°, 23.2°±0.2°, and 29.5°±0.2°.

[0007] In one embodiment, the crystalline form has an X-ray powder diffraction pattern comprising peaks at diffraction angles 2θ of 8.2±0.2°, 10.6±0.2°, 13.6±0.2°, 14.5±0.2°, 15.0±0.2°, 16.4±0.2°, 22.2±0.2°, 23.2±0.2° and 29.5±0.2°.

[0008] In one embodiment, the crystalline form has Figure 1 The X-ray powder diffraction pattern is shown.

[0009] In one embodiment, the crystalline form has a melting point of 231.3 ± 0.5 °C.

[0010] In a second aspect, the present application relates to a pharmaceutical composition comprising the crystal form of the first aspect of the present application.

[0011] In a third aspect, the present application relates to an ointment comprising the crystal form of the first aspect of the present application.

[0012] In one embodiment, the ointment comprises the crystalline form of the first aspect of the present application, a polyethylene glycol base and propylene glycol.

[0013] In one embodiment, based on the total weight of the ointment, the 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 accounts for 0.5%-3%, polyethylene glycol 400 accounts for 10%-50%, polyethylene glycol 3350 accounts for 30%-50%, and propylene glycol accounts for 20%-30%.

[0014] In one embodiment, based on the total weight of the ointment, the 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 accounts for 1.5%, polyethylene glycol 400 accounts for 33.5%, polyethylene glycol 3350 accounts for 40%, and propylene glycol accounts for 25%.

[0015] In a fourth aspect, the present application relates to use of the crystal form of the first aspect or the pharmaceutical composition of the second aspect in the preparation of a drug for treating JAK kinase-mediated diseases. BRIEF DESCRIPTION OF THE DRAWINGS

[0016] Figure 1 The XPRD spectrum of Form IV of Compound A is shown;

[0017] Figure 2 The DSC spectrum of Form IV of Compound A is shown;

[0018] Figure 3 The TGA spectrum of Form IV of Compound A is shown;

[0019] Figure 4 The XPRD spectrum of the crystalline form IIb of compound A is shown;

[0020] Figure 5 The DSC spectrum of the crystalline form IIb of compound A is shown;

[0021] Figure 6 The TGA spectrum of the crystalline form IIb of compound A is shown;

[0022] Figure 7 The XPRD pattern of the amorphous form of Compound A is shown;

[0023] Figure 8 The XPRD pattern of the comparative crystalline form of compound A is shown;

[0024] Fig. 9 The DSC spectrum of the comparative crystalline form of compound A is shown;

[0025] Fig.10 DVS of Form IV of Compound A is shown;

[0026] Fig.11 The DVS of Form IIb of Compound A is shown;

[0027] Fig.12 DVS showing the amorphous form of Compound A;

[0028] Fig.13 The cumulative permeation amounts of each of Prescription 1 and Prescription 2 within 24 hours are shown. DETAILED DESCRIPTION

[0029] The present application is further described in detail below through the accompanying drawings and embodiments. Through these descriptions, the characteristics and advantages of the present application will become clearer and more specific.

[0030] The word "exemplary" is used exclusively herein to mean "serving as an example, embodiment, or illustration." Any embodiment described herein as "exemplary" is not necessarily to be construed as preferred or advantageous over other embodiments. Although various aspects of the embodiments are shown in the drawings, the drawings are not necessarily drawn to scale unless otherwise noted.

[0031] In addition, the technical features involved in different embodiments of the present application described below can be combined with each other as long as they do not conflict with each other.

[0032] In the present application, the compound A referred to refers to 2-{3-[3-amino-4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]-1-(isopropylsulfonyl)azetidin-3-yl}acetonitrile, which can also be represented by the following structural formula:

[0033]

[0034] The synthesis method of compound A is disclosed in invention patent CN201711248509.8.

[0035] The term "substantially pure" as used herein when referring to a particular crystalline or amorphous form means that the crystalline or amorphous form contains less than 10% by weight, preferably less than 5% by weight, preferably less than 3% by weight, preferably less than 1% by weight of any other physical form of the compound.

[0036] As used herein, the term "substantially the same" when referring to X-ray diffraction peak positions means that typical peak position and intensity variability is taken into account. For example, those skilled in the art understand that peak positions (2θ) will show some variability, typically as much as 0.1 to 0.2 degrees, depending on the solvent used and the apparatus used to measure diffraction. Further, those skilled in the art understand that relative peak intensities will show instrument-to-instrument variability as well as variability caused by crystallinity, preferred orientation, prepared sample surface, and other factors known to those skilled in the art, and should be considered only as qualitative measurements.

[0037] In the present application, "at least one peak" means that there can be 1 peak, 2 peaks, 3 peaks or even all the peaks listed. For example, "at least one peak located at diffraction angles 2θ of 10.6°±0.2°, 15.0°±0.2° and 22.2°±0.2°" means "at diffraction angles 2θ of 10.6°±0.2°, 15.0°±0.2° and 22.2°±0.2°" 1 peak, 2 peaks or 3 peaks.

[0038] The abbreviations used in the present invention are explained as follows:

[0039] XRPD: X-ray powder diffraction

[0040] DSC: Differential Scanning Calorimetry

[0041] TGA: Thermogravimetric analysis

[0042] DVS: Dynamic Water Sorption

[0043] The X-ray powder diffraction pattern of the present invention is obtained by PANalytical Empyrean or X'Pert 3The X-ray powder diffractometer is used for the collection. The parameters of the X-ray powder diffraction method of the present invention are as follows:

[0044] X-ray source: Cu, Kα

[0045] 1.54060; 1.54439

[0046] Kα2 / Kα1 intensity ratio: 0.50

[0047] Voltage: 45 kilovolts (kV)

[0048] Current: 40 milliamperes (mA)

[0049] Scanning range: from 3.0 to 40.0 degrees.

[0050] The differential scanning calorimetry (DSC) graph of the present invention was collected on a TA Discovery 2500. The method parameters of the differential scanning calorimetry (DSC) of the present invention are as follows:

[0051] Scan rate: 10℃ / min

[0052] Shielding gas: N2.

[0053] The thermogravimetric analysis (TGA) graph of the present invention was collected on a TA Discovery 5500. The method parameters of the thermogravimetric analysis (TGA) of the present invention are as follows:

[0054] Scan rate: 10℃ / min

[0055] Shielding gas: N2.

[0056] The high performance liquid chromatography (HPLC) data in the present invention were collected from Agilent 1260, and the detector used was a diode array detector (DAD).

[0057] The HPLC method parameters for testing solubility of the present invention are as follows:

[0058] 1. Chromatographic column: ZORBAX Eclipse XDB-C18, 150×4.6mm, 5μm

[0059] 2. Mobile phase: A: 10mM NaH2PO4 aqueous solution (pH = 2.4)

[0060] B: Acetonitrile

[0061] The elution gradient is as follows:

[0062]

[0063]

[0064] 3. Flow rate: 0.85mL / min

[0065] 4. Injection volume: 5μL

[0066] 5. Detection wavelength: 225nm

[0067] 6. Column temperature: 35°C

[0068] 7. Diluent: ACN / mobile phase A1:1

[0069] Unless otherwise specified, the following examples were all operated at room temperature.

[0070] I. Crystalline and amorphous forms of compound A

[0071] Each crystalline form of Compound A can be characterized by one or more of the following: X-ray powder diffraction (X-ray diffraction peaks at different diffraction angles (2θ)), differential scanning calorimetry (DSC), thermogravimetric analysis (TGA), solubility, etc.

[0072] A. Form IV of Compound A

[0073] Form IV of compound A is anhydrous crystalline form which can be prepared according to the method described in Example 1.

[0074] pass Figure 1 The XPRD pattern shown is used to characterize Form IV of Compound A, which is also shown in Table 1.

[0075] Table 1

[0076]

[0077]

[0078] The crystal form IV can also be characterized by TGA and DSC. Figure 2 As shown, the melting temperature of the crystal form IV sample is 231.3°C. The TGA spectrum is as follows Figure 3 As shown, heating to 150°C shows a weight loss of 1.3%.

[0079] B. Crystalline Form IIb of the Compound

[0080] The crystalline form IIb of compound A can be prepared according to the method described in Example 2.

[0081] pass Figure 4 The XPRD pattern shown is used to characterize Form IIb of Compound A, which is also shown in Table 2.

[0082] Table 2

[0083]

[0084]

[0085] C. Amorphous form of compound A

[0086] The amorphous form of compound A can be prepared according to the method described in Example 3. Its XPRD spectrum is as follows Figure 7 As shown, the lack of sharp X-ray powder diffraction peaks indicates that it is amorphous.

[0087] II. Pharmaceutical compositions and uses

[0088] Compound A belongs to the JAK class of small molecule non-receptor tyrosine kinase inhibitors, which can be used to treat Janus kinase (JAK kinase) mediated diseases. For example, JAK kinase mediated diseases can be tumors or autoimmune diseases. Further, JAK kinase mediated diseases include but are not limited to tumor diseases such as leukemia, lymphoma, myeloproliferative tumors, myelodysplastic syndrome; also include but are not limited to rheumatoid arthritis, psoriatic arthritis, autoimmune skin diseases and other autoimmune diseases.

[0089] Furthermore, diseases mediated by JAK kinases may include, but are not limited to, autoimmune skin diseases such as psoriasis, atopic dermatitis, vitiligo, pruritus, scleroderma, alopecia areata, alopecia totalis, alopecia universalis, and androgenic alopecia.

[0090] The active agent of the present invention (i.e., a polymorph of Compound A herein or a solid form comprising two or more of the polymorphs) can be formulated into a pharmaceutical composition suitable for medical use in mammals. Any suitable route of administration can be used to provide an effective dose of any polymorph of Compound A to a patient. For example, the dosage form includes capsules, tablets, dispersions, suspensions, ointments, etc. In all dosage forms, the polymorphs of Compound A can be mixed with other suitable ingredients. The composition can be conveniently present in a unit dosage form and can be prepared by any method known in the pharmaceutical field. The pharmaceutical composition of the present invention comprises a therapeutically effective amount of an active agent and one or more inert pharmaceutically acceptable carriers, and optionally any other therapeutic ingredients, stabilizers or the like. From the perspective of compatibility with other ingredients of the preparation and not excessively damaging its receptor, the carrier must be pharmaceutically acceptable. The composition may further include diluents, buffers, binders, disintegrants, thickeners, lubricants, preservatives (including antioxidants), flavoring agents, taste masking agents, inorganic salts (e.g., sodium chloride), antimicrobial agents (e.g., benzalkonium chloride, etc.), sweeteners, antistatic agents, surfactants, sorbitan esters, lipids, etc. The active agents of the present invention may be formulated into compositions including those suitable for oral, rectal, topical, nasal, ocular, or parenteral (including intraperitoneal, intravenous, subcutaneous or intramuscular injection) administration.

[0091] In particular, the active agent of the present invention can be administered transdermally in the form of an ointment. The crystal form IV of compound A and the comparative crystal form ointment were prepared using the same formulation, and surprisingly, the crystal form IV ointment had a 24-hour cumulative permeation amount that was about 3 times higher than that of the comparative crystal form ointment, and the intradermal retention in the epidermis and dermis was about 3 to 5 times higher. As shown in Tables 8, 9 and Fig.13 shown.

[0092] Therefore, the present application also relates to an ointment, which contains the crystalline form IV described in the present application.

[0093] In one embodiment, the ointment comprises the crystalline form IV of compound A of the present application, a polyethylene glycol base and propylene glycol.

[0094] As the molecular weight of polyethylene glycol increases, its physical state can gradually transition from liquid to solid. Those skilled in the art can mix polyethylene glycols of different molecular weights in appropriate proportions to obtain a polyethylene glycol matrix with a suitable consistency. For example, a certain proportion of polyethylene glycol 400 and polyethylene glycol 3350 can be used to obtain a corresponding polyethylene glycol matrix.

[0095] In one embodiment, based on the total weight of the ointment, the 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 accounts for 0.5%-3%, polyethylene glycol 400 accounts for 10%-50%, polyethylene glycol 3350 accounts for 30%-50%, and propylene glycol accounts for 20%-30%.

[0096] In one embodiment, based on the total weight of the ointment, the 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 accounts for 1.5%, polyethylene glycol 400 accounts for 33.5%, polyethylene glycol 3350 accounts for 40%, and propylene glycol accounts for 25%.

[0097] The following examples will further illustrate the preparation of different polymorphs of the present invention, but are not intended to limit the present invention.

[0098] Example 1: Preparation and characterization of Form IV of Compound A

[0099] The crystalline form IV of compound A can be prepared according to the following process:

[0100] In a 250 mL flask, add 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, stir to dissolve, filter, filter out insoluble matter, add 250 mL of purified water to the filtrate, stir to crystallize, filter, and transfer the obtained solid to a 250 mL flask after drying, continue to add 200 mL of acetonitrile, heat and reflux for 3 h, cool, filter, and dry the obtained solid at 55-60 ° C to constant weight to obtain 18.0 g of off-white solid with a yield of 90.0%.

[0101] The crystal form IV was characterized by X-ray powder diffraction, TGA, and DSC. The X-ray powder diffraction data of the crystal form IV are shown in Table 1 and Figure 1 As shown; DSC spectrum as Figure 2 As shown, the melting temperature of the crystal form IV sample is 231.3°C; the TGA spectrum is as follows Figure 3 As shown, heating to 150°C shows a weight loss of 1.3%.

[0102] Example 2: Preparation and characterization of Form IIb

[0103] The crystalline form IIb of compound A can be prepared according to the following process:

[0104] In a 100 mL flask, add 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, stir to dissolve, filter to remove insoluble matter, add 50 mL of purified water to the filtrate, stir to crystallize, filter, and dry the resulting solid at 55-60°C to constant weight to obtain 5.6 g of an off-white solid with a yield of 88.9%.

[0105] The crystal form IIb was characterized by X-ray powder diffraction, TGA, and DSC. The X-ray powder diffraction data of the crystal form IIb are shown in Table 2 and Figure 4 The DSC spectrum is shown in Figure 5 As shown, the crystal form IIb has an endothermic peak between 50 and 200°C, and an endothermic peak near 221.5°C; the dehydration temperature is about 120.7°C, and the melting temperature is 220.2°C. The TGA spectrum is shown in Figure 6 As shown, it shows a weight loss of about 2.2% at 50-150°C.

[0106] Example 3: Preparation and characterization of amorphous form of compound A

[0107] Weigh 103.7 mg of compound A sample into a 50 mL flask, add 10 mL of acetone to dissolve, filter into another 50 mL flask with a PTFE filter membrane with a pore size of 0.45 μm, and rotary evaporate the solvent to obtain a solid.

[0108] Using Cu-Kα radiation, its X-ray powder diffraction pattern is as follows Figure 7 As shown, the lack of sharp X-ray powder diffraction peaks indicates that it is amorphous.

[0109] Comparative Example:

[0110] 2-(3-{3-(2,5-dioxopyrrolidin-1-yl)-4-[7-(hydroxymethyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl]-1H-pyrazol-1-yl}-1-(isopropylsulfonyl)azetidin-3-yl)acetonitrile 77.0g (0.15mol, 1.0eq) and 800ml of ethanol were added to a 2L reaction bottle. Under stirring, 92.0g (1.50mol, 10.0eq) of ethanolamine were added. The temperature was raised to reflux for 8h, the reaction was stopped, cooled, filtered, 500mL of water was added to the filter cake, stirred for crystallization, filtered, and the filter cake was vacuum dried at 60°C to obtain a light yellow solid. Weighing 55.6g, yield 92.4%.

[0111] The crystal form was characterized by X-ray powder diffraction and DSC. The X-ray powder diffraction data of the crystal form are as follows Figure 8 As shown, the DSC spectrum is Fig. 9As shown, the melting temperature of this crystalline form is 221.25°C.

[0112] Test Case

[0113] 1. Hygroscopicity

[0114] About 20-50 mg of each of the Form IV sample prepared in Example 1, the Form IIb sample prepared in Example 2, and the amorphous sample prepared in Example 3 were weighed for dynamic moisture sorption (DVS) testing, which was collected on the DVS Intrinsic Plus of SMS (Surface Measurement Systems).

[0115] The experimental results are shown in Table 3. The DVS of Form IV is as follows Fig.10 As shown, the DVS of Form IIb is as follows Fig.11 As shown, amorphous DVS Fig.12 As shown, Form IV and Form IIb showed no crystalline change after DVS test, and the amorphous sample turned into Form IIb after the test.

[0116] Table 3

[0117]

[0118] Compared with the amorphous form, the crystalline form IV of the present invention has a smaller moisture absorption weight gain as the humidity increases and better physical stability.

[0119] 2. Comparative study of solid-state physical and chemical stability

[0120] About 2-3 mg of the crystalline form IV (initial purity 99.87%) obtained in Example 1 of the present invention was weighed, placed in an oven at 80°C with the mouth closed, and sampled for XRPD and HPLC after 1 day. The crystalline form IV was placed in a stabilization box at 25°C / 60%RH and 40°C / 75%RH with the mouth open, and sampled for XRPD and HPLC after 1 month.

[0121] About 2 to 3 mg of the crystalline form IIb (initial purity 99.84%) obtained in Example 2 of the present invention was weighed, placed in an oven at 80°C with the lid closed, and sampled for XRPD and HPLC after 1 day. The sample was placed in a stabilization box at 25°C / 60%RH and 40°C / 75%RH with the lid open, and sampled for XRPD and HPLC after 1 month.

[0122] Weigh about 2-3 mg of the amorphous form (initial purity 99.74%) obtained in Example 3 of the present invention, place it in an oven at 80°C with the lid closed, take a sample after 1 day to measure XRPD and HPLC, and place it in a stabilization box at 25°C / 60%RH and 40°C / 75%RH with the lid open, take a sample after 1 month to measure XRPD and HPLC.

[0123] The experimental results are shown in Table 4. The results show that the crystal form and purity of Form IV did not change significantly after being placed under 25°C / 60%RH, 40°C / 75%RH and 80°C. The crystal form and purity of Form IIb did not change significantly after being placed under 25°C / 60%RH, 40°C / 75%RH and 80°C. After the amorphous form was placed under 25°C / 60%RH, 40°C / 75%RH and 80°C, the crystal form did not change significantly, but the purity decreased slightly under 40°C / 75%RH and 80°C.

[0124] Compared with the amorphous form, the crystal form IV of the present invention has better physical / chemical stability under the conditions of 25°C / 60% RH and 40°C / 75% RH. During storage and use, it can remain stable without specific temperature and humidity control conditions, and can well meet the requirements of drug storage and use.

[0125] Table 4

[0126]

[0127] *Relative purity = test sample HPLC purity / starting sample HPLC purity × 100%

[0128] 3. Solubility comparison study

[0129] The crystal form IV sample prepared in Example 1, the crystal form IIb sample prepared in Example 2, and the amorphous sample prepared in Example 3 were weighed, and prepared into a suspension with H2O, simulated body fluid (SBF), propylene glycol and PEG-400, and stirred and mixed at 25°C with a feed concentration of 10 mg / mL (for the PEG-400 system, the feed concentration was adjusted to >150 mg / mL), and the solubility of each sample was measured at a point taken after 24 hours. After sampling, centrifugation (12000 rpm, 2 min) and filtration (0.45 μm PTFE membrane) were performed, and the HPLC concentration of the filtrate was determined, and the solid sample after centrifugation was tested by XRPD.

[0130] The experimental results are shown in Table 5. The results show that the solubility of each sample in water and SBF is low, less than 0.0041 mg / mL, and the solubility in PEG-400 is higher than 42 mg / mL. Most systems have undergone crystal form changes after the solubility test: Crystal form IIb is converted to crystal form I in the propylene glycol system; Crystal form IV is converted to crystal form IIb in water and SBF systems; amorphous is converted to crystal form IIb in water and SBF, to crystal form I in propylene glycol, and to a mixed crystal form of crystal form I and crystal form IIb in PEG-400.

[0131] Compared with the amorphous form, the crystalline form IV of the present invention has comparable solubility in solution systems, and even has better solubility in propylene glycol and SBF.

[0132] Table 5

[0133]

[0134] *“No” means that no crystal transformation occurs and the original crystal form remains.

[0135] 4. Comparative study of particle size distribution

[0136] Take about 10-30 mg of each of the Form IV sample prepared in Example 1, the Form IIb sample prepared in Example 2, and the amorphous sample prepared in Example 3, and then add about 5 mL of Isopar G (containing 0.2% lecithin). After the samples to be tested are fully mixed, they are added to the injection system so that the shading degree reaches an appropriate range. The experiment is started, and the particle size distribution is tested before and 30 seconds after ultrasound.

[0137] The results are shown in Table 6. Compared with the amorphous form, the experimental results show that the crystalline form IIb and the crystalline form IV of the present invention have a more uniform particle size distribution (the particle size distribution SD is smaller) than the amorphous form.

[0138] Table 6

[0139]

[0140] * Refers to data before ultrasound

[0141] ** Refers to the data after 30W ultrasound for 30 seconds

[0142] 5. Comparative study of crystal habit

[0143] Weigh about 2 mg of the Form IV sample prepared in Example 1, the Form IIb sample prepared in Example 2, and the amorphous sample prepared in Example 3, respectively, and place them on glass slides. Add a small amount of vacuum silicone oil to disperse the samples, then cover them with coverslips and observe them under a polarizing microscope.

[0144] The experimental results show that Form IIb is a plate-like crystal, Form IV is a long rod-shaped crystal, and amorphous is an irregular particle. The self-regulation of crystals helps to remove impurities and improve product purity, while the regular geometric shape is a macroscopic manifestation of the self-regulation of crystals. Therefore, in terms of crystal habits, Form IV has more advantages than amorphous.

[0145] 6. Comparative study of wettability

[0146] About 200 mg of each of the Form IV sample prepared in Example 1, the Form IIb sample prepared in Example 2, and the amorphous sample prepared in Example 3 were weighed, and each of the relevant solid-state samples was pressed into tablets, and their contact angles to water were measured.

[0147] The results are shown in Table 7. Compared with the amorphous form, the experimental results show that the crystal form IV of the present invention has a smaller contact angle and is more easily wetted by water.

[0148] Table 7

[0149] Material Contact angle Form IIb 62.5～69.2° Form IV 55.3～60.9° Amorphous 80.0～83.3°

[0150] Pharmaceutical composition

[0151] According to the prescription ratio listed in Table 8, different types of compound A were weighed into PEG 400 and propylene glycol, stirred and dispersed for 15 to 30 minutes, and then PEG 3350 was added and stirred for about 1 hour to prepare an anhydrous ointment.

[0152] Table 8 Compound A ointment prescription list

[0153] Raw materials Prescription 1 (%) Prescription 2 (%) Compound A (Form IV) 1.5 / Compound A (Comparative Example Crystal Form) / 1.5 PEG 400 33.5 33.5 Propylene glycol 25 25 PEG 3350 40 40 total 100 100

[0154] The skin of 1-3 month old Bama miniature pigs was selected as the skin for in vitro transdermal experiments to test the penetration of compound A ointment with different crystal forms in the skin. Specifically, a Franz diffusion cell was selected, the receiving cell volume was 12 mL, the inner diameter was 1.5 cm, and the receiving liquid was normal saline. The temperature was set to 32 ° C, and about 20 mg of the ointment was placed on the skin of the miniature pigs. The sampling points were 4, 8, 12, 16 and 24 hours. After 24 hours, the skin was removed, the remaining drug on the skin was collected, and the skin was layered and sampled for extraction. After the sample was filtered, the content was determined by LC-MS / MS.

[0155] Fig.13 The cumulative permeation within 24 hours of each of Prescription 1 and Prescription 2 is shown, and the intradermal retention (μg) is given in Table 9. Surprisingly, it was found that the 24-hour cumulative permeation of the crystal form IV ointment was about 3 times higher than that of the comparative crystal form ointment, and the intradermal retention in the epidermis and dermis was about 3 to 5 times higher.

[0156] Table 9 In vitro transdermal test of compound A ointment Intradermal retention (μg)

[0157] Prescription 1 Prescription 2 Stratum corneum 98.57 19.83 Epidermis 4.26 0.73 Dermis 2.11 0.84

[0158] The present application has been described above in conjunction with preferred embodiments, but these embodiments are only exemplary and serve only as an illustration. On this basis, various replacements and improvements may be made to the present application, all of which fall within the scope of protection of the present application.

Claims

1. A 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, wherein: The crystalline form has an X-ray powder diffraction pattern including peaks located at diffraction angles 2θ of 8.2°±0.2°, 14.5°±0.2° and 16.4°±0.2°.

2. The crystal form according to claim 1, wherein The X-ray powder diffraction pattern of the crystalline form further includes at least one peak located at diffraction angles 2θ of 10.6°±0.2°, 15.0°±0.2° and 22.2°±0.2°.

3. The crystal form according to claim 1 or 2, wherein The X-ray powder diffraction pattern of the crystalline form further includes at least one peak located at diffraction angles 2θ of 13.6°±0.2°, 23.2°±0.2° and 29.5°±0.2°.

4. The crystal form according to claim 1, wherein The crystalline form has an X-ray powder diffraction pattern including peaks at diffraction angles 2θ of 8.2±0.2°, 10.6±0.2°, 13.6±0.2°, 14.5±0.2°, 15.0±0.2°, 16.4±0.2°, 22.2±0.2°, 23.2±0.2° and 29.5±0.2°.

5. The crystal form according to claim 1, wherein The crystal form has an X-ray powder diffraction pattern as shown in FIG1 .

6. The crystal form according to claim 1, wherein The crystalline form has a melting point of 231.3±0.5°C.

7. A pharmaceutical composition comprising the crystal form according to any one of claims 1 to 6.

8. An ointment comprising the crystal form according to any one of claims 1 to 6.

9. The ointment according to claim 8, wherein The ointment comprises the crystal form according to any one of claims 1 to 6, a polyethylene glycol base and propylene glycol.

10. The ointment according to claim 9, wherein Based on the total weight of the ointment, the 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 accounts for 0.5%-3%, polyethylene glycol 400 accounts for 10%-50%, polyethylene glycol 3350 accounts for 30%-50%, and propylene glycol accounts for 20%-30%.

11. The ointment according to claim 10, wherein Based on the total weight of the ointment, the 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 accounts for 1.5%, polyethylene glycol 400 accounts for 33.5%, polyethylene glycol 3350 accounts for 40%, and propylene glycol accounts for 25%.

12. Use of the crystal form according to any one of claims 1 to 6, the pharmaceutical composition according to claim 7, or the ointment according to any one of claims 8 to 11 in the preparation of a medicament for treating a JAK kinase-mediated disease.

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