Crystal form, preparation method and application of a pyrazolyl-amino-pyrimidinyl derivative

Abstract

The present invention discloses a crystalline form, preparation method, and application of a pyrazolyl-amino-pyrimidinyl derivative. The present invention provides a crystalline form I of compound 1, as well as its preparation method and application. This crystalline form exhibits one or more of the following advantages: excellent physical and chemical properties, solid-state stability, good solubility, low hygroscopicity, and good processability in formulation processes.

Description

Technical Field

[0001] The present invention relates to a crystal form, a preparation method and application of a pyrazolyl-amino-pyrimidinyl derivative. Background Art

[0002] The JAK-STAT pathway mediates the intracellular signaling of multiple cytokines in the body. Previous studies have shown that elevated levels of multiple inflammatory cytokines, such as Th1 (γ-interferon), Th2 (IL-4, IL-13, IL-31), and Th22 (IL-22), are present in the skin lesions of AD patients, suggesting that the pathogenesis of AD is closely related to the JAK-STAT pathway. IL-4 and IL-13 bind to IL-4 receptors α and γ and IL-13 receptor α1, activating JAK1 / 3 and promoting STAT3 / 6 phosphorylation. STAT3 can disrupt skin barrier integrity by downregulating proteins associated with KC differentiation; STAT6 can upregulate chemokines involved in the pathogenesis of AD, and Th0 cells differentiate into Th2 cells through the JAK1 / 3-STAT6 pathway, leading to the onset of AD. Eosinophils are one of the most important effector cells in AD. IL-5 and its receptor β chain complex participate in the regulation of eosinophil proliferation, survival, and efficacy through the JAK2-STAT1 / 5 pathway. Activated eosinophils are attracted to the skin by chemokines involved in the pathogenesis of AD released by epidermal cells in a high Th2 immune environment, further aggravating the AD condition.

[0003] Among the JAK inhibitors currently under development, those making rapid progress include Japan Tobacco's Delgocitinib (already marketed), Incyte's Ruxolitinib (already marketed), and Pfizer's Tofacitinib (Phase II). Results showed that Delgocitinib significantly improved clinical scores in patients in a Phase III clinical study for the treatment of AD. Phase II and Phase III studies of Ruxolitinib for the treatment of AD demonstrated rapid antipruritic and anti-inflammatory effects and good tolerability. In a Phase II study of Tofacitinib, 2% ointment significantly improved EASI scores in AD patients after four weeks of treatment, demonstrating good local tolerability and safety. In addition, domestic JAK inhibitors under development for the treatment of AD, including Suzhou Zejing Bio's Jaktinib Hydrochloride Cream and Jiangsu Hengrui's SHR0302 Alkaline Ointment, have also entered Phases I / II and II / III, respectively. However, current local treatments for AD still cannot meet clinical needs. Continuing to develop safer and more effective JAK inhibitors can improve the current situation of insufficient treatment drugs and enhance the treatment effect and quality of life of patients with atopic dermatitis. It has great significance and market prospects.

[0004] Preclinical studies have found that LNK01004 (its structure is shown below) is a pan-Janus kinase (JAK) inhibitor with strong inhibitory effects on JAK1, JAK2, and TYK2, and can simultaneously inhibit multiple cytokine-induced p-STAT signaling pathways in vitro and in vivo. In vitro and in vivo experimental results show that LNK01004 can inhibit the cytokine-induced p-STAT signaling pathway in immune cells. Skin application can also effectively inhibit the p-STAT signaling pathway induced by cytokines associated with psoriasis or atopic dermatitis in skin tissue. Unlike Ruxolitinib and Tofacitinib, LNK01004 can also inhibit the proliferation of keratinocytes.

[0005] Summary of the Invention

[0006] The present invention provides a crystalline form, preparation method and application of a pyrazolyl-amino-pyrimidinyl derivative. The crystalline form satisfies one or more of the following advantages: good physical and chemical properties, solid-state stability, good solubility, low hygroscopicity and good processability in formulation.

[0007] The present invention provides a crystalline form I of compound 1;

[0008]

[0009] The crystalline form I uses Cu-Kα radiation, and the X-ray powder diffraction pattern expressed in 2θ has diffraction peaks at the following positions: 8.60°±0.2°, 10.25°±0.2°, 11.96°±0.2°, 14.35°±0.2°, 15.39°±0.2°, 16.59°±0.2°, 17.06°±0.2° and 18.16°±0.2°.

[0010] In a certain embodiment, the X-ray powder diffraction pattern of the crystalline form I expressed in 2θ angles further has diffraction peaks at one or more of the following positions: 12.77°±0.2°, 13.48°±0.2°, 14.04°±0.2°, 17.27°±0.2°, 18.83°±0.2°, 20.52°±0.2°, 20.77°±0.2°, 21.45°±0.2°, 22.12°±0.2°, 22.79°±0.2°, 23.55°±0.2°, 24.04°±0.2°, 24.40°±0.2°, 25.08°±0.2°, 25.87°±0.2°, 26.51°±0.2°, 26.73°±0.2°, 26.89°±0.2°, 27.36°±0.2° and 28.29°±0.2°.

[0011] In one embodiment, the X-ray powder diffraction pattern of the crystalline form I expressed in 2θ angles further has diffraction peaks at one or more of the following positions: 28.93°±0.2°, 29.42°±0.2°, 30.63°±0.2°, 33.00°±0.2°, 33.37°±0.2°, 34.43°±0.2° and 37.09°±0.2°.

[0012] In one embodiment, the X-ray powder diffraction pattern of the crystalline form I expressed in 2θ angles has diffraction peaks as shown in the following table:

[0013]

[0014]

[0015] In one embodiment, the X-ray powder diffraction pattern of the crystalline form I expressed in 2θ angles is substantially as follows Figure 1 shown.

[0016] In one embodiment, the differential scanning calorimetry diagram of the crystalline form I has an endothermic peak at 207.4°C to 209.2°C.

[0017] In one embodiment, the differential scanning calorimetry diagram of the crystalline form I has an endothermic peak at 207.4°C to 209.2°C, and the heat of melting is 123.76 J / g.

[0018] In one embodiment, the differential scanning calorimetry diagram of the crystalline form I is substantially as follows: Figure 2 shown.

[0019] In one embodiment, the thermogravimetric analysis of the Form I ranges from 30.07° C. to 208.96° C., with a weight loss of 0.0%, indicating that the Form I is an anhydrous compound.

[0020] In one embodiment, the thermogravimetric analysis diagram of the crystalline form I is substantially as follows: Figure 3 shown.

[0021] The present invention also provides a method for preparing the above-mentioned crystal form I, which is Scheme 1 or Scheme 2;

[0022] Scheme 1 comprises the following steps: crystallizing a methanol solution of compound 1 to obtain the crystalline form I;

[0023]

[0024] Scheme 2 comprises the following steps: cooling a solution of compound 1 in tetrahydrofuran / methanol and isopropanol to obtain the crystalline form I.

[0025] In one embodiment, in embodiment 1, during the crystallization, the mass of the solution is (1:6)-(1:8.5) of the mass of the compound 1; preferably (1:6.3)-(1:8.3).

[0026] In one embodiment, the first embodiment preferably comprises the following steps: adding methanol to a solution of compound 1 and tetrahydrofuran at 40° C., concentrating the solution, adding methanol again to the concentrate, stirring, and crystallizing to obtain the crystalline form I;

[0027] The temperature at which the compound 1 is dissolved in the tetrahydrofuran to form a solution is preferably 50-60°C;

[0028] The mass ratio of the compound 1 to the tetrahydrofuran is preferably (1:8)-(1:9); more preferably 1:8.7;

[0029] The mass ratio of the compound 1 to the first added methanol is preferably (1:16)-(1:18); more preferably 1:17;

[0030] The mass of the concentrated solution is preferably 4-6 times the mass of the compound 1, more preferably 5 times;

[0031] The mass ratio of the compound 1 to the second added methanol is preferably (1:2)-(1:3); more preferably 1:2.3.

[0032] In one embodiment, the first embodiment may further include the following post-processing steps: filtration, washing, drying under reduced pressure, and sieving to obtain the crystal form I.

[0033] In one embodiment, in embodiment 2, the mass ratio of compound 1 to tetrahydrofuran / methanol is (1:3)-(1:5); preferably 1:4.

[0034] In one embodiment, in embodiment 2, the mass ratio of tetrahydrofuran to methanol in the tetrahydrofuran / methanol is (2:1)-(1:2); preferably 1:1.

[0035] In one embodiment, in embodiment 2, the mass ratio of compound 1 to isopropanol is (1:11)-(1:13); preferably 1:12.

[0036] In one embodiment, in embodiment 2, the temperature at which compound 1 is dissolved in tetrahydrofuran / methanol and isopropanol is 50-60°C; preferably 55°C.

[0037] In one embodiment, in embodiment 2, the cooling is to reduce the temperature to 0-5°C, preferably to 0°C.

[0038] In one scheme, in scheme 2, the holding time after cooling is related to the reaction scale, and the reaction end point is generally when the product no longer increases; the holding time is preferably 15-30 hours, more preferably 24 hours.

[0039] In one embodiment, the second embodiment preferably includes the following operations: heating compound 1 and tetrahydrofuran / methanol, adding isopropanol, dissolving, adding isopropanol again, cooling, forming a suspension, and obtaining the crystalline form I.

[0040] In one embodiment, the second embodiment may further include the following post-processing steps: filtration, washing, drying under reduced pressure, and sieving to obtain the crystalline form I.

[0041] The present invention also provides a pharmaceutical composition comprising the above-mentioned crystal form I and pharmaceutical excipients.

[0042] The present invention also provides a use of the above-mentioned crystal form I in the preparation of a drug for treating and / or preventing diseases associated with JAK kinase.

[0043] In one embodiment, the disease associated with JAK kinase is inflammatory bowel disease, psoriasis, vitiligo, atopic dermatitis, systemic lupus erythematosus, asthma, diabetic nephropathy, chronic myeloid leukemia (CML), essential thrombocythemia (ET), polycythemia vera (PV), myelofibrosis (MF), breast cancer or ovarian cancer.

[0044] Without violating the common sense in the art, the above-mentioned preferred conditions can be arbitrarily combined to obtain preferred embodiments of the present invention.

[0045] The reagents and raw materials used in the present invention are commercially available.

[0046] The positive progress of the present invention is that: Form I has good physical and chemical properties, good high temperature (e.g., 60°C) and high humidity (92.5% RH) stability, high pressure (10 MPa) solid-state stability, good solubility (much greater than 8 μg / mL), low hygroscopicity, uniform particle size distribution, good solid form and formulation processability, and good development prospects. BRIEF DESCRIPTION OF THE DRAWINGS

[0047] Figure 1 This is the XRPD spectrum of Form I.

[0048] Figure 2 This is the DSC spectrum of Form I.

[0049] Figure 3 This is the TGA spectrum of Form I.

[0050] Figure 4 This is the PLM spectrum of Form I.

[0051] Figure 5 This is the SEM spectrum of Form I.

[0052] Figure 6A and Figure 6B This is the DVS spectrum of Form I.

[0053] Figure 7 This is the XRPD spectrum of Form I before and after DVS testing.

[0054] Figure 8 This is the XRPD spectrum of Form II.

[0055] Figure 9 This is the DSC spectrum of Form II.

[0056] Figure 10 This is the TGA spectrum of Form II.

[0057] Figure 11 This is the PLM spectrum of Form II.

[0058] Figure 12 This is the XRPD spectrum of Form III.

[0059] Figure 13 This is the DSC spectrum of Form III.

[0060] Figure 14 This is the TGA spectrum of Form III.

[0061] Figure 15 This is the PLM spectrum of Form III.

[0062] Figure 16 This is the XRPD spectrum of Form IV.

[0063] Figure 17 This is the DSC spectrum of Form IV.

[0064] Figure 18 This is the TGA spectrum of Form IV.

[0065] Figure 19 This is the PLM spectrum of Form IV.

[0066] Figure 20 This is the XRPD spectrum of Form V.

[0067] Figure 21 This is the DSC spectrum of Form V.

[0068] Figure 22 This is the TGA spectrum of Form V.

[0069] Figure 23 This is the PLM spectrum of Form V.

[0070] Figure 24 This is the XRPD spectrum of Form VI.

[0071] Figure 25 This is the DSC spectrum of Form VI.

[0072] Figure 26 This is the TGA spectrum of Form VI.

[0073] Figure 27 This is the PLM spectrum of Form VI.

[0074] Figure 28 The XRPD spectrum of amorphous form.

[0075] Figure 29 The DSC spectrum of amorphous. DETAILED DESCRIPTION

[0076] The present invention is further illustrated by way of examples below, but the present invention is not limited to the scope of the examples. Experimental methods in the following examples where specific conditions are not specified were performed according to conventional methods and conditions, or selected according to the product specifications.

[0077] XRPD analysis method

[0078] 1. Test Preparation

[0079] category name Remark instrument Bruker D8 Advance X-ray Diffractometer or equivalent instrument Material Monocrystalline silicon panels or equivalent board

[0080] 2. Parameter setting

[0081]

[0082]

[0083] Note: The above parameters are established based on Bruker D8 XRPD and can be adjusted for different instruments.

[0084] 3. Testing and result reporting

[0085] Load an appropriate amount of sample (e.g., 20-50 mg, adjustable) onto the single crystal silicon plate and spread the sample evenly across the center of the plate, as shown in the figure below. If the sample contains large particles, a back-mounted sample plate can be used. If response intensity is not required, both flat and grooved single crystal silicon plates can be used; otherwise, a grooved plate should be used to maintain a consistent loading height.

[0086] If necessary, apply a thin layer of petroleum jelly or silicone oil to the surface of the single crystal silicon plate to adhere the sample, and gently tap off any excess sample. Load the sample plate into the XRPD sample holder, scan and acquire the spectrum, and report the results.

[0087] DSC test method

[0088] 1. Test Preparation

[0089]

[0090] 2. Parameter setting

[0091] parameter set up Nitrogen flow rate 50ml / min Data collection frequency 1.00s / pt Temperature range Room temperature to 300℃ Heating rate 10℃ / min

[0092] 3. Testing and result reporting

[0093] Take an appropriate amount of sample (do not overfill to prevent overflow during heating) and place it in an augmentation tray. Cover and seal with a capping device. Use an empty augmentation tray as a blank control. The tray and lid for the blank control should match those for the sample. Install the trays in the appropriate sample racks: the sample augmentation tray on the sample rack and the blank augmentation tray on the control rack. Select the method, use the workstation software for data processing, and report the results.

[0094] TGA test methods

[0095] 1. Test Preparation

[0096] category name Remark instrument Thermogravimetric analyzer TA TGA55 / GA550 or equivalent Material Sample tray Aluminum or platinum

[0097] 2. Parameter setting

[0098] parameter set up Balance purge flow rate 40ml / min Furnace purge flow rate 10ml / min Temperature range Room temperature to 300℃ Heating rate 10℃ / min

[0099] 3. Testing and result reporting

[0100] Place the empty sample pan in the target position on the Auto Sampler. Click "TARE" on the workstation. The instrument will automatically attach the pan, turn off the furnace, and tare. Accurately weigh approximately 2-10 mg of sample and place it on the tareed sample pan. Edit the sample information, select the method, and click "Sta" to begin sample analysis. The workstation will automatically record the curve of the sample weight percentage changing with temperature. Click "Analysis" and select "Weight change" from the drop-down menu. Click "Analyze" and the workstation will automatically calculate the sample weight loss percentage (%) and report the results.

[0101] PLM (Polarized Light Microscopy) Test Method

[0102] 1. Test Preparation

[0103] category name instrument Nikon LV100POL polarizing microscope Material Slides / cover slips

[0104] 2. Testing and result reporting

[0105] Place several particles of the sample in mineral oil (e.g., silicone oil) to form a suspension and place on a clean glass slide. Place an appropriate amount of the suspension on a glass slide and cover with a coverslip. For irregularly shaped particles, the particle size characterization must also include particle size information. The homogeneity of the powder should be verified using an appropriate magnification. Report the results of the micrographs.

[0106] Preparation and Characterization Example 1 of Form I:

[0107] LNK01004 (prepared by referring to Example 113 of CN113227074A, amorphous) (net content: 1.73 kg, 1.00 ± 0.02X) and tetrahydrofuran (15 kg, 8.7X) were added to reactor R1, the temperature was raised to 50-60°C, and stirred for 1-3 hours until all dissolved.

[0108] The temperature was controlled at 40°C, and methanol (30 kg, 17X) was added. After concentrating under reduced pressure to 4.0-6.0X, methanol (4 kg, 2.3X) was added and stirred for 1-3 hours.

[0109] The suspension was filtered and methanol (2 kg, 1.2X) was added to wash the filter cake. The mixture was dried under reduced pressure at 40-50°C until the moisture and residual solvent content were within the acceptable range (tetrahydrofuran ≤ 720 ppm, methanol ≤ 5000 ppm). After drying, 1.362 kg of the final product, LNK01004, Form I (JR-C200212007-FPF21001), was obtained by sieving. The XRPD pattern is shown in FIG. Figure 1 As shown, the purity was 99.91% and the yield was 90%.

[0110] XRPD data of JR-C200212007-FPF21001

[0111]

[0112]

[0113]

[0114] Characterization of Form I

[0115] Typical characterization data collected for Form I obtained from JR-C200212007-FPF21001 are shown below. These data demonstrate that Form I is a stable, solvent-free crystalline product with very high purity, a stable melting point, solvent-free encapsulation, good morphology and particle size distribution, and no significant hygroscopicity. These properties are beneficial for the subsequent development and production of APIs and formulations.

[0116] Characterization data of Form I

[0117]

[0118] Preparation Example 2 of Form I:

[0119] To reactor R1, LNK01004 (prepared with reference to Example 113 of CN113227074A, amorphous) (net content: 5 kg, 1.00 ± 0.02X) and tetrahydrofuran / methanol = 1:1 (20 kg, 4.0X) were added, the temperature was raised to 50-60°C, isopropanol (5 kg, 1.0X) was added, and the mixture was stirred for 1-3 hours until completely dissolved.

[0120] The temperature was controlled at 55° C., and isopropyl alcohol (55 kg, 11.0X) was added simultaneously and maintained for 1 to 3 hours.

[0121] The reaction temperature was lowered to 0°C over 5.0 hours and maintained at 0°C for 24 hours to form a suspension.

[0122] The suspension was filtered and isopropyl alcohol (10 kg, 2.0X) was added to wash the filter cake.

[0123] The mixture was dried under reduced pressure at 40-50°C until the water content and residual solvent content were within acceptable limits (tetrahydrofuran ≤ 720 ppm, methanol ≤ 5000 ppm, isopropanol ≤ 5000 ppm). After drying, the mixture was sieved to obtain 4.7 kg of the final product, LNK01004 Form I (characterization data of which were consistent with those in Example 1 for the preparation of Form I), with a purity of 100.0% and a yield of 94%.

[0124] Crystal form I effect example 1:

[0125] Water activity experiment at 1.25℃

[0126] 20 mg of Form I was weighed, 1 mL of acetone / water system with different water activities was added, and the mixture was stirred at 25° C. for 10, 12, or 22 days. The obtained solid was filtered and characterized by XRPD.

[0127] Water activity experiment at 25℃

[0128]

[0129] The results of this water activity experiment show that anhydrous Form I is stable over a wide range of water activities (1%-100%) for a long period of time (up to 22 days) without converting into hydrates or other crystalline forms. This helps the formulation maintain a stable crystalline form after ingestion, facilitating temperature-dependent absorption and drug exposure.

[0130] 2. Stability test results

[0131] Crystal form I was placed under high temperature 60 ° C and high humidity 92.5% RH conditions for 30 days, and placed under light conditions 1×ICH (total illumination not less than 1.2×10 6 Lux·hr, near-ultraviolet energy not less than 200w·hr / m 2 ) and accelerated (40±2℃ / 75%±5%RH) storage for 6 months and long-term (25±2℃ / 60%±5%RH) storage for 18 months, the appearance, related substances, content on an anhydrous and solvent-free basis, moisture, crystal form, content and microbial limit results did not change.

[0132] Stability test

[0133]

[0134] Example 2 of the effect of Form I: Dynamic solubility study of Form I

[0135] Approximately 20 mg of Form I was weighed and placed in a 40 mL glass vial. 10 mL of simulated gastrointestinal fluid was added and stirred at 400 rpm at 37°C. Approximately 1 mL of the suspension was removed at 1 hour, 4 hours, and 24 hours, and centrifuged at 37°C to determine the solubility of Form I at each time point. After 24 hours, the pH of the suspension was measured, and the remaining suspension was centrifuged for XRPD characterization of the remaining solid.

[0136] Study on the dynamic solubility (37°C) of Form I

[0137]

[0138] The results showed that Form I had good solubility in simulated gastrointestinal fluid (far greater than 8 μg / mL), and the preparation could maintain stable absorption during subsequent formulation development and production.

[0139] Effect of Form I Example 3: Pressure Test of Form I

[0140] Approximately 10 mg of Form I (sample number: FR00970-12-SU1) was weighed and tableted using a hydraulic press at 10 MPa for 5 minutes. XRPD analysis was then performed to investigate the changes in crystallinity and crystallinity. The results showed that the dominant Form I remained stable under high pressure (10 MPa), facilitating subsequent formulation stability production.

[0141] Study on crystal transformation behavior under pressure

[0142] pressure XRPD Remark 10Mpa Form I The crystal form and crystallinity remain unchanged

[0143] Example 4 of the effect of Form I: Simulated dry grinding of Form I

[0144] Approximately 10 mg of Form I (sample number: FR00970-12-SU1) was weighed and ground in a mortar for 3 minutes. XRPD analysis was then performed to investigate the crystal transformation and crystallinity changes. The results showed that Form I remained stable under dry grinding conditions, facilitating subsequent formulation production.

[0145] Simulated dry grinding experiment

[0146]

[0147]

[0148] Example 5: Simulated dry grinding of Form I

[0149] Approximately 10 mg of Form I (sample number: FR00970-12-SU1) was weighed and added with 40 μL of water or ethanol, respectively. The sample was then ground in a mortar for 3 minutes. XRPD analysis was performed to investigate the crystal transformation and crystallinity changes. The results showed that the dominant Form I remained stable under wet grinding conditions, facilitating subsequent formulation production.

[0150] Simulated wet grinding experiment

[0151] solvent XRPD Remark ethanol Form I The crystal form remains unchanged, but the crystallinity is slightly reduced. water Form I The crystal form remains unchanged, but the crystallinity is slightly reduced.

[0152] Effect of Form I Example 6: Pharmacodynamic Data of Form I

[0153] This study used a migration assay to determine the half-inhibitory concentration (IC50) values ​​of Form I of the compound LNK01004, ruxolitinib, tofacitinib, and upadacitinib against JAK1, JAK2, JAK3, and TYK2 kinase activity. In the assay, Form I of the compound LNK01004, ruxolitinib, tofacitinib, and upadacitinib were tested against JAK1, JAK2, JAK3, and TYK2 kinases at a starting concentration of 10 μM, using a 3-fold serial dilution series for a total of 10 concentrations in duplicate wells. The ATP concentration was 1 mM. The test results are shown in the table below:

[0154] The half-inhibitory concentration (IC50) value of Form I on the kinase activity of JAK1, JAK2, JAK3, and TYK2

[0155]

[0156] The experimental results show that Form I of the compound LNK01004 has stronger kinase activity inhibition ability in JAK1, JAK2, JAK3, and TYK2 activity tests than ruxolitinib, tofacitinib, and upadacitinib, demonstrating that Form I of LNK01004 has better pharmaceutical properties at the same concentration and dosage.

[0157] Comparative Example 1 Preparation and Characterization of Form II

[0158] Approximately 50 mg of LNK01004 (prepared as amorphous form, as described in Example 113 of CN113227074A) was weighed and thoroughly dissolved in 2 ml of acetone / water (v:v = 1:1) at 50°C. The resulting solution was filtered through a 0.45 μm filter to obtain a clear solution. The resulting clear solution was cooled to 5°C at a cooling rate of 0.1°C / min. The resulting solid was collected by filtration to obtain Form II. Form II (sample number: FR00970-7-SC12) contained 0.3% residual acetone and a water content of 12.2%. Form II has a relatively low dehydration temperature of T onset 56.8℃, it is a highly crystalline metastable hydrate.

[0159] Characterization data of Form II

[0160]

[0161] Comparative Example 2 Preparation and Characterization of Form III

[0162] Approximately 50 mg of LNK01004 (prepared as amorphous according to Example 113 of CN113227074A) was weighed into a 2 mL glass vial. 1 mL of methanol / dichloromethane (v:v = 1:1) was added and the mixture was suspended at 50°C and 400 rpm for one week. The resulting suspension was filtered, and the solid fraction was characterized to yield Form III. Form III (sample number: FR00970-7-SC6) contains no residual solvent and a water content of 6.6%, demonstrating its high crystallinity as a metastable hydrate.

[0163] Characterization data of Form III

[0164]

[0165] Comparative Example 3 Preparation and Characterization of Form IV

[0166] About 50 mg of LNK01004 (prepared with reference to Example 113 of CN113227074A, amorphous) was weighed and placed in a 2 mL glass bottle. 1 mL of tetrahydrofuran / water (v:v = 1:1) solvent was added and suspended at 25 ° C and 400 rpm for one week. The resulting suspension was filtered and the resulting solid was characterized, which was Form IV ( Figure 24 Form IV (sample number: FR00970-12-SU3) contained 3% residual tetrahydrofuran and 5.4% water. Form IV transformed into Form VI after 2 days of storage at ambient temperature (20-25°C, 80-95% RH), indicating that it is a highly crystalline metastable hydrate of Form IV.

[0167] Characterization data of Form IV

[0168]

[0169] Comparative Example 4 Preparation and Characterization of Form V

[0170] Approximately 20 mg of LNK01004 (prepared as amorphous form, as described in Example 113 of CN113227074A) was weighed and thoroughly dissolved in 1 ml of DMF / n-heptane (v:v = 1:1). The solution was then filtered through a 0.45 μm filter to obtain a clear solution. The resulting clear solution was then slowly evaporated at room temperature to yield Form V (sample number: FR00970-11-VD3), a solid containing 1.6 equivalents of DMF. Upon heating to 150°C and removing the solvent, Form V transformed into Form I (whose XRPD data were consistent with those in Example 1 for the preparation of Form I).

[0171] Characterization data of Form V

[0172]

[0173]

[0174] “ / ”: Indicates not done.

[0175] Comparative Example 5 Preparation and Characterization of Form VI

[0176] Approximately 50 mg of LNK01004 (prepared as amorphous, as described in Example 113 of CN113227074A) was weighed and placed in a 2 mL glass vial. 1 mL of a tetrahydrofuran / water (v:v = 1:1) solvent was added and the mixture was suspended at 25°C and 400 rpm for one week. The resulting suspension was filtered, and the resulting solid was characterized, revealing Form IV. Form IV crystallized to Form VI after 2 days under ambient conditions (20-25°C, 80-95% RH). Form VI (sample number: FR00970-9-TC17) contained 2.3% residual THF and a water content of 4.8%. Form VI is unstable and can transform into other forms under certain conditions. Exposure to zero humidity resulted in Form VII, which then transformed to Form III upon heating to 120°C and cooling to room temperature.

[0177] Characterization data of Form VI

[0178]

[0179] Comparative Example 6 Preparation and Characterization of Form VII

[0180] Approximately 50 mg of LNK01004 (prepared with reference to Example 113 of CN113227074A, amorphous) was weighed into a 2 mL glass vial, and 1 mL of tetrahydrofuran / water (v:v = 1:1) was added. The mixture was suspended at 25°C and 400 rpm for one week. The resulting suspension was filtered, and the resulting solid was characterized, revealing Form IV. Form IV transformed into Form VII after equilibration at 0% RH for 12 hours. Form VII was only stable under low RH conditions; within 2 hours of returning to 60% RH, Form VII transformed into Form III. Form VII and Form III had similar XRPD spectra, except for slight shifts in the positions of several peaks.

[0181] Comparative Example 7 Preparation and Characterization of Amorphous

[0182] Approximately 50 mg of LNK01004 (prepared as amorphous form, as described in Example 113 of CN113227074A) was weighed and placed in a 2 mL glass vial. 1 mL of acetonitrile / water (v:v = 1:1) was added and thoroughly dissolved at 50°C. The resulting clear solution was filtered through a 0.45 μm filter to obtain a clear solution. The clear solution was cooled to 5°C at a rate of 0.1°C / min. The resulting solid was collected by filtration to obtain the amorphous form (sample number: FR00970-7-SC8). DSC thermal analysis revealed the crystalline form (heating from 30°C to melt at a rate of 10°C / min; cooling from melt to -20°C at a rate of 20°C / min): the amorphous form was desolvated between 30°C and 110°C. Further heating from 140°C to 190°C resulted in conversion to Form I (characterization data similar to those in "Form I Preparation and Characterization Example 1").

[0183] Characterization data of crystalline amorphous form

[0184]

[0185] Although the above describes specific embodiments of the present invention, it should be understood by those skilled in the art that these are merely illustrative and that various changes or modifications may be made to these embodiments without departing from the principles and essence of the present invention. Therefore, the scope of protection of the present invention is defined by the appended claims.

Claims

1. A crystalline form I of compound 1, characterized in that: The crystalline form I uses Cu-Kα radiation, and the X-ray powder diffraction pattern expressed in 2θ has diffraction peaks at the following positions: 8.60°±0.2°, 10.25°±0.2°, 11.96°±0.2°, 14.35°±0.2°, 15.39°±0.2°, 16.59°±0.2°, 17.06°±0.2° and 18.16°±0.2°; 2. The crystalline form I according to claim 1, wherein The crystalline form I satisfies one or more of the following conditions: (1) The X-ray powder diffraction pattern of the crystalline form I expressed in 2θ angles further has diffraction peaks at one or more of the following positions: 12.77°±0.2°, 13.48°±0.2°, 14.04°±0.2°, 17.27°±0.2°, 18.83°±0.2°, 20.52°±0.2°, 20.77°±0.2°, 21.45°±0.2°, 22 .12°±0.2°, 22.79°±0.2°, 23.55°±0.2°, 24.04°±0.2°, 24.40°±0.2°, 25.08°±0.2°, 25.87°±0.2°, 26.51°±0.2°, 26.73°±0.2°, 26.89°±0.2°, 27.36°±0.2°, and 28.29°±0.2°; (2) The differential scanning calorimetry of the crystalline form I has an endothermic peak at 207.4°C to 209.2°C; (3) The thermogravimetric analysis of the crystal form I shows a weight loss of 0.0% from 30.07°C to 208.96°C.

3. The crystalline form I according to claim 2, wherein The X-ray powder diffraction pattern of the crystalline form I expressed in 2θ angles also has diffraction peaks at one or more of the following positions: 28.93°±0.2°, 29.42°±0.2°, 30.63°±0.2°, 33.00°±0.2°, 33.37°±0.2°, 34.43°±0.2° and 37.09°±0.2°.

4. The crystalline form I according to claim 3, wherein The crystalline form I satisfies one or more of the following conditions: (1) The X-ray powder diffraction pattern of the crystalline form I expressed at 2θ angles has the diffraction peaks shown in the following table: (2) The differential scanning calorimetry of the Form I has an endothermic peak at 207.4°C to 209.2°C, and a heat of fusion of 123.76 J / g; (3) The thermogravimetric analysis diagram of the crystal form I is basically as shown in Figure 3.

5. The crystalline form I according to claim 4, wherein The crystalline form I satisfies one or more of the following conditions: (1) The X-ray powder diffraction pattern of the crystalline form I expressed at 2θ angles is substantially as shown in FIG1 ; (2) The differential scanning calorimetry diagram of the crystal form I is basically as shown in Figure 2.

6. A method for preparing the crystalline form I according to any one of claims 1 to 5, characterized in that: It is Option 1 or Option 2; wherein, Scheme 1 comprises the following steps: crystallizing a methanol solution of compound 1 to obtain the crystalline form I; Scheme 2 comprises the following steps: cooling a solution of compound 1 in tetrahydrofuran / methanol and isopropanol to obtain the crystalline form I.

7. The method for preparing the crystal form I according to claim 6, wherein: The preparation method meets one or more of the following conditions: (1) In the first embodiment, during the crystallization, the mass of the solution is (1:6)-(1:8.5) of the mass of the compound 1; (2) The first scheme comprises the following steps: adding methanol to a solution of compound 1 and tetrahydrofuran at 40°C, concentrating the solution, adding methanol again to the concentrate, stirring, and crystallizing to obtain the crystalline form I; (3) The first solution further includes the following post-processing steps: filtration, washing, drying under reduced pressure, and sieving to obtain the crystal form I; (4) In Scheme 2, the mass ratio of compound 1 to tetrahydrofuran / methanol is (1:3)-(1:5); (5) In the second scheme, the mass ratio of tetrahydrofuran to methanol in the tetrahydrofuran / methanol is (2:1)-(1:2); (6) In Scheme 2, the mass ratio of compound 1 to isopropanol is (1:11)-(1:13); (7) In Scheme 2, the temperature at which compound 1 is dissolved in tetrahydrofuran / methanol and isopropanol is 50–60 °C; (8) In the second solution, the temperature is lowered to 0-5°C; (9) In the second scheme, the holding time after cooling is 15-30h; (10) The second scheme also includes the following post-processing steps: filtration, washing, drying under reduced pressure, and sieving to obtain the crystal form I.

8. The method for preparing the crystal form I according to claim 7, wherein: The preparation method meets one or more of the following conditions: (1) In the first embodiment, during the crystallization, the mass of the solution is (1:6.3)-(1:8.3) of the mass of the compound 1; (2) In scheme 1, the temperature at which the compound 1 is dissolved in the tetrahydrofuran to form a solution is 50-60°C; (3) In Scheme 1, the mass ratio of the compound 1 to the tetrahydrofuran is (1:8)-(1:9); (4) In Scheme 1, the mass ratio of the compound 1 to the first added methanol is (1:16)-(1:18); (5) In scheme 1, the mass of the concentrated solution is 4-6 times the mass of the compound 1; (6) In scheme 1, the mass ratio of the compound 1 to the second added methanol is (1:2)-(1:3); (7) In Scheme 2, the mass ratio of compound 1 to tetrahydrofuran / methanol is 1:4; (8) In the second scheme, the mass ratio of tetrahydrofuran to methanol in the tetrahydrofuran / methanol is 1:1; (9) In Scheme 2, the mass ratio of compound 1 to isopropanol is 1:12; (10) In Scheme 2, the temperature at which compound 1 is dissolved in tetrahydrofuran / methanol and isopropanol is 55 °C; (11) In the second solution, the temperature is lowered to 0°C; (12) In the second scheme, the heat preservation time after cooling is 24 hours.

9. The method for preparing the crystal form I according to claim 8, wherein: The preparation method meets one or more of the following conditions: (1) In Scheme 1, the mass ratio of the compound 1 to the tetrahydrofuran is 1:8.7; (2) In scheme 1, the mass ratio of the compound 1 to the first added methanol is 1:17; (3) In scheme 1, the mass of the concentrated solution is 5 times the mass of the compound 1; (4) In Scheme 1, the mass ratio of the compound 1 to the methanol added for the second time is 1:2.

3.

10. The method for preparing the crystal form I according to claim 6, wherein: The second scheme comprises the following operations: heating compound 1 and tetrahydrofuran / methanol, adding isopropanol, dissolving, adding isopropanol again, cooling, forming a suspension, and obtaining the crystalline form I.

11. A pharmaceutical composition comprising the crystalline form I according to any one of claims 1 to 5 and a pharmaceutical excipient.

12. Use of the crystalline form I according to any one of claims 1 to 5 in the preparation of a drug for treating and / or preventing diseases associated with JAK kinase; The disease associated with JAK kinase is inflammatory bowel disease, psoriasis, vitiligo, atopic dermatitis, systemic lupus erythematosus, asthma, diabetic nephropathy, chronic myeloid leukemia, essential thrombocythemia, polycythemia vera, myelofibrosis, breast cancer or ovarian cancer.

13. Use of the crystalline form I according to any one of claims 1 to 5 in the preparation of a drug for treating and / or preventing a disease, characterized in that: The disease is inflammatory bowel disease, psoriasis, vitiligo, atopic dermatitis, systemic lupus erythematosus, asthma, diabetic nephropathy, chronic myeloid leukemia, essential thrombocythemia, polycythemia vera, myelofibrosis, breast cancer or ovarian cancer.

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