Salt form, crystal form of JAK inhibitor, preparation method and application thereof

Abstract

The present invention provides a salt form, a crystal form, a preparation method and an application of a JAK inhibitor. The crystal form A of the compound represented by formula I has a characteristic diffraction peak at the following 2θ angles in its X-ray powder diffraction pattern: 5.9°±0.2°, 7.4°±0.2°, 11.6°±0.2°, 21.7°±0.2°, and 23.8°±0.2°.

Description

[0001] This application claims priority to PCT patent application PCT / CN2020 / 109279 filed on August 14, 2020 and PCT patent application PCT / CN2021 / 075086 filed on February 3, 2021. The entire text of the above patent applications is incorporated herein by reference. Technical Field

[0002] The present invention relates to a salt form, a crystal form of a JAK inhibitor, and a preparation method and application thereof. Background Art

[0003] The Janus kinase (JAK) signaling pathway found in interferon-induced receptor-mediated gene expression has been shown to be a common signaling pathway used by many cytokines and growth factors. The mammalian JAK family of intracellular tyrosine kinases has four members: Janus kinase 1 (JAK1), Janus kinase 2 (JAK2), Janus kinase 3 (JAK3), and tyrosine kinase 2 (TYK2). JAKs range in size from 120 to 140 kDa and contain seven conserved JAK homology (JH) domains that define this superfamily of kinases.

[0004] Each JAK isoform can be used by multiple cytokine pathways, and even the biological activity of many cytokines can be regulated by inhibition of single or multiple JAKs. Inhibition of JAKs can be used to prevent, inhibit or treat the development or onset of various diseases and disorders, including hyperproliferative diseases and cancers, such as leukemias and lymphomas, immune and inflammatory disorders, such as transplant rejection, asthma, chronic obstructive pulmonary disease, allergies, rheumatoid arthritis, psoriasis, atopic dermatitis, Crohn's disease, ulcerative colitis, amyotrophic lateral sclerosis and multiple sclerosis.

[0005] WO2016119700A1 discloses that the compound represented by formula I is an effective JAK inhibitor for preventing or treating one or more symptoms of a Janus kinase-mediated disease;

[0006]

[0007] The solubility of the free base form of the compound shown in Formula I in water is too low, which will affect its dissolution and absorption in the body, resulting in too low bioavailability, making it unsuitable for further drug development. At the same time, due to its low water solubility, it is not easy to be purified during the production process, which brings certain difficulties to industrial production. Summary of the invention

[0008] The present invention provides a salt form, a crystal form, a preparation method and an application of a JAK inhibitor. The salt form and the crystal form have good JAK kinase inhibitory activity, solubility, stability and bioavailability, and enhance the developability of an oral preparation of the compound shown in Formula I.

[0009] The present invention provides a crystalline form A of a hydrochloride hydrate of a compound represented by formula I, wherein the X-ray powder diffraction (XRPD) spectrum thereof has characteristic diffraction peaks at the following 2θ angles: 5.9°±0.2°, 7.4°±0.2°, 11.6°±0.2°, 21.7°±0.2° and 23.8°±0.2°;

[0010]

[0011] In some embodiments of the present invention, the X-ray powder diffraction pattern of the crystalline form A has characteristic diffraction peaks at the following 2θ angles: 5.9°±0.2°, 7.4°±0.2°, 11.6°±0.2°, 17.7°±0.2°, 17.8°±0.2°, 21.7°±0.2°, 23.6°±0.2°, 23.8°±0.2°, 29.0°±0.2°, 30.4°±0.2° and 34.9°±0.2°.

[0012] In some embodiments of the present invention, the X-ray powder diffraction pattern of the crystalline form A has characteristic diffraction peaks at the following 2θ angles: 5.9°±0.2°, 7.4°±0.2°, 8.9°±0.2°, 11.6°±0.2°, 14.6°±0.2°, 17.7°±0.2°, 17.8°±0.2°, 18.7°±0.2°, 1 9.5°±0.2°, 21.0°±0.2°, 21.7°±0.2°, 21.9°±0.2°, 22.6°±0.2°, 23.6°±0.2°, 23.8°±0.2°, 29.0°±0.2°, 30.4°±0.2°, 33.3°±0.2°, 34.9°±0.2° and 37.7°±0.2°.

[0013] In some embodiments of the present invention, the X-ray powder diffraction pattern of the crystalline form A has characteristic diffraction peaks at the following 2θ angles: 5.9°±0.2°, 6.6°±0.2°, 7.4°±0.2°, 8.9°±0.2°, 10.5°±0.2°, 11.6°±0.2°, 12.3°±0.2°, 13.9°±0.2°, 14.2°±0.2°, 14.6°±0.2°, 17.1°±0.2°, 17.7°±0.2°, 17.8°±0.2°, 18.7°±0.2°, 19.5°±0.2°, 20.4°±0.2°, 21.0°±0.2°, 21.2°±0.2°, 21.7°±0.2°, 22. .2°, 21.9°±0.2°, 22.6°±0.2°, 23.6°±0.2°, 23.8°±0.2°, 24.9°±0.2°, 26.6°±0.2°, 26.8°±0.2°, 28.3°±0.2°, 29.0°±0.2°, 30.4°±0.2°, 31.2°±0.2 °, 32.3°±0.2°, 33.3°±0.2°, 34.2°±0.2°, 34.9°±0.2°, 35.6°±0.2°, 35.9°±0.2°, 36.4°±0.2°, 36.9°±0.2°, 37.7°±0.2°, 39.3°±0.2° and 39.8°±0.2°.

[0014] In some embodiments of the present invention, the X-ray powder diffraction pattern of the crystalline form A has characteristic diffraction peaks at the following 2θ angles: 5.9°, 6.6°, 7.4°, 8.9°, 10.5°, 11.6°, 12.3°, 13.9°, 14.2°, 14.6°, 17.1°, 17.7°, 17.8°, 18.7°, 19.5°, 20.4°, 21.0°, 21.2°, 21.7°, 21.9°, 22.6°, 23.6°, 23.8°, 24.9°, 26.6°, 26.8°, 28.3°, 29.0°, 30.4°, 31.2°, 32.3°, 33.3°, 34.2°, 34.9°, 35.6°, 35.9°, 36.4°, 36.9°, 37.7°, 39.3° and 39.8°.

[0015] In some embodiments of the present invention, the XRPD pattern analysis data of the crystalline form A is shown in Table 1:

[0016] Table 1. X-ray powder diffraction pattern analysis data of Form A

[0017] Diffraction angle (°2θ) Relative strength [%] Diffraction angle (°2θ) Relative strength [%] 5.9 20.7 23.6 15.3 6.6 0.9 23.8 29.8 7.4 55.4 24.9 14.3 8.9 7.8 26.6 1.2 10.5 1.3 26.8 3.9 11.6 100 28.3 4.2 12.3 0.7 29.0 19.0 13.9 3.6 30.4 19.7 14.2 2.0 31.2 0.8 14.6 6.2 32.3 1.2 17.1 3.9 33.3 8.0 17.7 17.0 34.2 1.7 17.8 18.8 34.9 15.6 18.7 5.0 35.6 3.1 19.5 14.8 35.9 2.3 20.4 1.7 36.4 0.8 21.0 9.4 36.9 0.9 21.2 2.6 37.7 5.7 21.7 33.6 39.3 1.0 21.9 11.8 39.8 2.5 22.6 5.3 .

[0018] In some embodiments of the present invention, the XRPD pattern of the crystalline form A is as follows Figure 1 shown.

[0019] In some embodiments of the present invention, the thermogravimetric analysis curve of the crystalline form A has a weight loss of 8.31±0.50% before 148°C; and a weight loss of 4.76±0.50% at 148°C-228°C.

[0020] In some embodiments of the present invention, the thermogravimetric analysis curve of the crystalline form A has a weight loss of 8.31% before 148°C and a weight loss of 4.76% at 148°C-228°C.

[0021] In some embodiments of the present invention, the thermogravimetric analysis curve of the crystalline form A has a weight loss of 8.3143% before 148°C and a weight loss of 4.7637% at 148°C-228°C.

[0022] In some embodiments of the present invention, the thermogravimetric analysis curve of the crystal form A is as follows Figure 3 shown.

[0023] In some embodiments of the present invention, the thermogravimetric analysis curve is obtained by detection under the conditions of a temperature increase range of 10°C-300°C and a temperature increase rate of 10°C / min.

[0024] In some embodiments of the present invention, the differential scanning calorimetry (DSC) analysis chart of the crystalline form A has absorption peaks at 165°C±3°C and 198°C±3°C, respectively.

[0025] In some embodiments of the present invention, the differential scanning calorimetry diagram of the crystalline form A has absorption peaks at 165.14° C. and 197.70° C., respectively.

[0026] In some embodiments of the present invention, the differential scanning calorimetry diagram of the crystalline form A has absorption peaks with onset temperatures of 141°C±3°C and 179°C±3°C, respectively.

[0027] In some embodiments of the present invention, the differential scanning calorimetry diagram of the crystalline form A has absorption peaks with onset temperatures of 140.82° C. and 179.20° C., respectively.

[0028] In some embodiments of the present invention, the differential scanning calorimetry diagram of the crystalline form A is as follows: Figure 2 shown.

[0029] In some embodiments of the present invention, the differential scanning calorimetry diagram is obtained under the conditions of a temperature increase range of 25°C-300°C and a temperature increase rate of 10°C / min.

[0030] In some embodiments of the present invention, the crystalline form A is a single crystal.

[0031] In some embodiments of the present invention, the single crystal of Form A belongs to the monoclinic system, P21 / c space group.

[0032] In some embodiments of the present invention, the unit cell parameters of the crystalline form A are α=90°, β=101.262(3)°, γ=90°.

[0033] In some embodiments of the present invention, the unit cell volume of the crystalline form A is

[0034] The unit cell parameters and / or unit cell volume of the single crystal of the crystal form A can be obtained by X-ray single crystal diffraction detection. The X-ray wavelength λ of the X-ray single crystal diffraction can be

[0035] In some embodiments of the present invention, in the crystalline form A, the molar ratio of the compound of formula I, HCl and water is 1:x:y, wherein x is greater than 0 but not greater than 3, and y is greater than 0 but not greater than 3.

[0036] In some embodiments of the present invention, in the crystalline form A, the molar ratio of the compound of formula I, HCl and water is 1:2:2.

[0037] The present invention also provides a method for preparing a crystalline form A of a hydrochloride hydrate of a compound shown in Formula I, comprising the following steps: precipitating crystals from a hydrochloride solution of a compound shown in Formula I, wherein the crystals are Form A; wherein the hydrochloride solution of the compound shown in Formula I contains a hydrochloride of the compound shown in Formula I, an organic solvent and water, and the organic solvent is selected from one or a mixture of two or more of methanol, ethanol, n-propanol, n-butanol, isopropanol, isobutanol and tert-butanol. The two or more herein should be understood to include two. It should be clear to those skilled in the art that the hydrochloride solution of the compound shown in Formula I does not contain other organic solvents, unless it is an unavoidable impurity or solvent residue.

[0038] Wherein, in the hydrochloride of the compound of formula I, the molar ratio of the compound of formula I to HCl is 1:x, where x is greater than 0 and not greater than 3, for example, 1:2.

[0039] In some embodiments, the organic solvent is ethanol.

[0040] In some embodiments, the volume ratio of the organic solvent to water is 5-15:0.5-1.5, for example, 10:1 to 6:1, for example, 10:1 to 8:1, for example, 9:1.

[0041] In some embodiments, the temperature of the hydrochloride solution of the compound of Formula I is 30-70°C.

[0042] In some embodiments, the step of precipitating crystals from the hydrochloride solution of the compound of Formula I comprises: cooling the hydrochloride solution of the compound of Formula I, for example, cooling to 20°C to 30°C.

[0043] In some embodiments, the step of precipitating crystals from the hydrochloride solution of the compound of Formula I comprises: stirring the hydrochloride solution of the compound of Formula I at 20°C to 30°C to precipitate crystals; the stirring time is, for example, 48-96 hours, and for example, 48 hours.

[0044] In some embodiments, the preparation method further comprises: after crystals are precipitated from the hydrochloride solution of the compound represented by Formula I, filtering, washing and drying the obtained filter cake to obtain Form A.

[0045] In some embodiments, the preparation method further comprises: cooling the solution of the crystalline form A of the hydrochloride hydrate of the compound shown in Formula I for crystallization to obtain a single crystal of the crystalline form A of the hydrochloride hydrate of the compound shown in Formula I, wherein the solvent of the solution of the crystalline form A of the hydrochloride hydrate of the compound shown in Formula I is a mixture of ethanol and water; for example, the temperature of the solution of the crystalline form A of the hydrochloride hydrate of the compound shown in Formula I is 55-75°C, for example, 60-70°C, and for example, 64-66°C; for example, the volume ratio of ethanol to water in the mixture of ethanol and water is 5-15:0.5-1.5, for example, 10:1 to 6:1, for example, 10:1 to 8:1, and for example, 9:1; for example, the cooling is cooling to 20°C to 30°C.

[0046] In some embodiments, the hydrochloride solution of the compound of Formula I is obtained by mixing raw materials including the hydrochloride hydrate of the compound of Formula I, an organic solvent and water; for example, the hydrochloride solution of the compound of Formula I is a mixture of the hydrochloride hydrate of the compound of Formula I, an organic solvent and water. For example, the volume ratio of the organic solvent to water is 5-15:0.5-1.5, for example, 10:1 to 6:1, for example, 10:1 to 8:1, for example, 9:1; for example, the amount ratio of the hydrochloride hydrate of the compound of Formula I to water is 250-450 mg:1 mL, for example, 300-400 mg:1 mL, for example, 350 mg:1 mL.

[0047] In some embodiments, the hydrochloride solution of the compound of formula I is obtained by mixing raw materials including the compound of formula I, an organic solvent, water and a concentrated hydrochloric acid solution. For example, the hydrochloride solution of the compound of formula I is a mixture of the compound of formula I, an organic solvent, water and a concentrated hydrochloric acid solution. For example, the preparation method further includes: mixing the compound of formula I, an organic solvent and water, heating the resulting mixture to 30-70° C., adding a concentrated hydrochloric acid solution, and obtaining the hydrochloride solution of the compound of formula I.

[0048] Wherein, the ratio of the concentrated hydrochloric acid solution to the compound represented by formula I can be, for example, 0.38mL-0.57mL: 1g, for example, 0.4mL-0.5mL: 1g, and for example, 0.38mL-0.40mL: 1g, 0.40mL-0.42mL: 1g, 0.42mL-0.44mL: 1g, 0.44mL-0.46mL: 1g, 0.46mL-0.48mL: 1g, 0.48mL-0.50mL: 1g, 0.50mL-0.52mL: 1g, 0.52mL-0.54mL: 1g or 0.54mL-0.57mL: 1g.

[0049] The ratio of the organic solvent to the compound represented by formula I may be, for example, 5 mL-15 mL: 1 g, for example, 8 mL-12 mL: 1 g, and for example, 5 mL-6 mL: 1 g, 6 mL-7 mL: 1 g, 7 mL-8 mL: 1 g, 8 mL-9 mL: 1 g, 9 mL-10 mL: 1 g, 10 mL-11 mL: 1 g, 11 mL-12 mL: 1 g, 12 mL-13 mL: 1 g, 13 mL-14 mL: 1 g or 14 mL-15 mL: 1 g.

[0050] The volume ratio of the organic solvent to water may be, for example, 5-15:0.5-1.5, for example, 10:1 to 6:1, for example, 10:1 to 8:1, and for example, 9:1.

[0051] The ratio of water to the compound represented by formula I may be, for example, 0.5 mL-1.5 mL: 1 g; for example, 0.8 mL-1.2 mL: 1 g, and for example, 0.5 mL-0.6 mL: 1 g, 0.6 mL-0.7 mL: 1 g, 0.7 mL-0.8 mL: 1 g, 0.8 mL-0.9 mL: 1 g, 0.9 mL-1.0 mL: 1 g, 1.0 mL-1.1 mL: 1 g, 1.1 mL-1.2 mL: 1 g, 1.2 mL-1.3 mL: 1 g, 1.3 mL-1.4 mL: 1 g or 1.4 mL-1.5 mL: 1 g.

[0052] The ratio of the organic solvent, water and the compound represented by formula I may be, for example, 5 mL-15 mL: 0.5 mL-1.5 mL: 1 g, for example, 8 mL-12 mL: 0.8 mL-1.2 mL: 1 g, and for example, 9 mL: 1 mL: 1 g.

[0053] The ratio of the organic solvent, water, concentrated hydrochloric acid solution and the compound represented by formula I can be, for example, 5 mL-15 mL: 0.5 mL-1.5 mL: 0.38 mL-0.57 mL: 1 g, for example, 8 mL-12 mL: 0.8 mL-1.2 mL: 0.38 mL-0.48 mL: 1 g, and for example, 9 mL: 1 mL: 0.41 mL: 1 g.

[0054] The concentration of the concentrated hydrochloric acid solution may be, for example, 8 mol / L-12 mol / L, for example, 10 mol / L-12 mol / L, for example, 12 mol / L.

[0055] In the hydrochloride solution of the compound represented by formula I, the molar ratio of HCl to the compound represented by formula I can be, for example, 2:1 to 3:1, for example, 2.0:1 to 2.5:1, and for example, 2.1:1 to 2.3:1.

[0056] In some embodiments, the hydrochloride solution of the compound represented by Formula I is obtained by mixing raw materials including a solution of the compound represented by Formula I and a concentrated hydrochloric acid solution; for example, the hydrochloride solution of the compound represented by Formula I is a mixture of raw materials consisting of a solution of the compound represented by Formula I and a concentrated hydrochloric acid solution.

[0057] For example, the solvent of the solution of the compound represented by formula I is a mixture of an organic solvent and water; for another example, the solution of the compound represented by formula I is a mixture of the compound represented by formula I, an organic solvent and water.

[0058] For example, the volume ratio of the organic solvent to water is 5-15:0.5-1.5, for example 10:1 to 6:1, for example 10:1 to 8:1, for example 9:1.

[0059] For example, the solution of the compound represented by formula I is obtained by mixing the compound represented by formula I and the solvent at a temperature of 30-70°C, and the temperature is, for example, 40-60°C, and for example, 45-55°C.

[0060] The present invention also provides a method for preparing a crystalline form A of a hydrochloride hydrate of a compound shown in Formula I, comprising the following steps: precipitating crystals from a hydrochloride solution of a compound shown in Formula I, wherein the crystals are Form A; wherein the hydrochloride solution of the compound shown in Formula I contains the compound shown in Formula I, an organic solvent, water and a concentrated hydrochloric acid solution, and the organic solvent is selected from one or a mixture of two or more of methanol, ethanol, n-propanol, n-butanol, isopropanol, isobutanol and tert-butanol. The two or more herein should be understood to include two. It should be clear to those skilled in the art that the hydrochloride solution of the compound shown in Formula I does not contain other organic solvents, unless it is an unavoidable impurity or solvent residue.

[0061] In some embodiments, the organic solvent is ethanol.

[0062] In some embodiments, the hydrochloride solution of the compound of Formula I is a mixture of the compound of Formula I, an organic solvent, water, and a concentrated hydrochloric acid solution.

[0063] In some embodiments, the ratio of the concentrated hydrochloric acid solution to the compound of Formula I is 0.38 mL-0.57 mL: 1 g, for example, 0.4 mL-0.5 mL: 1 g, and for example, 0.38 mL-0.40 mL: 1 g, 0.40 mL-0.42 mL: 1 g, 0.42 mL-0.44 mL: 1 g, 0.44 mL-0.46 mL: 1 g, 0.46 mL-0.48 mL: 1 g, 0.48 mL-0.50 mL: 1 g, 0.50 mL-0.52 mL: 1 g, 0.52 mL-0.54 mL: 1 g or 0.54 mL-0.57 mL: 1 g.

[0064] In some embodiments, the ratio of the organic solvent to the compound of Formula I is 5 mL-15 mL: 1 g, for example, 8 mL-12 mL: 1 g, for example, 5 mL-6 mL: 1 g, 6 mL-7 mL: 1 g, 7 mL-8 mL: 1 g, 8 mL-9 mL: 1 g, 9 mL-10 mL: 1 g, 10 mL-11 mL: 1 g, 11 mL-12 mL: 1 g, 12 mL-13 mL: 1 g, 13 mL-14 mL: 1 g or 14 mL-15 mL: 1 g.

[0065] In some embodiments, the ratio of water to the compound of formula I is 0.5mL-1.5mL:1g; for example, 0.8mL-1.2mL:1g, and for example, 0.5mL-0.6mL:1g, 0.6mL-0.7mL:1g, 0.7mL-0.8mL:1g, 0.8mL-0.9mL:1g, 0.9mL-1.0mL:1g, 1.0mL-1.1mL:1g, 1.1mL-1.2mL:1g, 1.2mL-1.3mL:1g, 1.3mL-1.4mL:1g or 1.4mL-1.5mL:1g.

[0066] In some embodiments, the volume ratio of the organic solvent to water is 5-15:0.5-1.5, preferably 10:1 to 6:1, more preferably 10:1 to 8:1, and more preferably 9:1.

[0067] In some embodiments, the ratio of the organic solvent, water and the compound of formula I is 5 mL-15 mL: 0.5 mL-1.5 mL: 1 g, preferably 8 mL-12 mL: 0.8 mL-1.2 mL: 1 g, and further preferably 9 mL: 1 mL: 1 g.

[0068] In some embodiments, the ratio of the organic solvent, water, concentrated hydrochloric acid solution and the compound represented by Formula I is 5 mL-15 mL: 0.5 mL-1.5 mL: 0.38 mL-0.57 mL: 1 g, preferably 8 mL-12 mL: 0.8 mL-1.2 mL: 0.38 mL-0.48 mL: 1 g, and further preferably 9 mL: 1 mL: 0.41 mL: 1 g.

[0069] In some embodiments, the concentration of the concentrated hydrochloric acid solution is 8 mol / L-12 mol / L, preferably 10 mol / L-12 mol / L, and more preferably 12 mol / L.

[0070] In some embodiments, in the hydrochloride solution of the compound of formula I, the molar ratio of HCl to the compound of formula I is 2:1 to 3:1, preferably 2.0:1 to 2.5:1, and further preferably 2.1:1 to 2.3:1.

[0071] In some embodiments, the hydrochloride solution of the compound of Formula I is a mixture of a solution of the compound of Formula I and a concentrated hydrochloric acid solution.

[0072] In some embodiments, the hydrochloride solution of the compound of Formula I is obtained by adding (eg, by dropwise addition) a concentrated hydrochloric acid solution to a solution of the compound of Formula I.

[0073] In some embodiments, the solvent of the solution of the compound of Formula I is a mixture of an organic solvent and water.

[0074] In some embodiments, the solution of the compound of formula I is obtained by mixing the compound of formula I and the solvent at a temperature of 30-70°C, preferably 40-60°C, and further preferably 45-55°C.

[0075] In some embodiments, the solution of the compound of Formula I is a mixture of the compound of Formula I, an organic solvent and water.

[0076] In some embodiments, in the solvent of the solution of the compound of Formula I, the volume ratio of the organic solvent to water is 10:1 to 6:1, preferably 10:1 to 8:1, and more preferably 9:1.

[0077] In some embodiments, the preparation method further comprises: mixing the compound represented by formula I, an organic solvent and water, heating the resulting mixture to 30-70°C (preferably 40-60°C, more preferably 45-55°C), adding (for example, by dropwise addition) a concentrated hydrochloric acid solution to obtain a hydrochloride solution of the compound represented by formula I.

[0078] In some embodiments, the preparation method further comprises: stirring the hydrochloride solution of the compound of formula I at 30-70° C. The step of stirring the hydrochloride solution of the compound of formula I at 30-70° C. is performed before the step of precipitating crystals from the hydrochloride solution of the compound of formula I. The stirring time is, for example, 1-5 hours, and for example, 1 hour.

[0079] In some embodiments, the step of precipitating crystals from the hydrochloride solution of the compound of Formula I comprises: cooling the hydrochloride solution of the compound of Formula I, preferably, the cooling is to 20°C to 30°C.

[0080] In some embodiments, the step of precipitating crystals from the hydrochloride solution of the compound of Formula I comprises: stirring the hydrochloride solution of the compound of Formula I at 20° C. to 30° C. to precipitate crystals. The stirring time is, for example, 48-96 hours, and for example, 48 hours.

[0081] In some embodiments, the preparation method further comprises: after crystals are precipitated from the hydrochloride solution of the compound represented by Formula I, filtering, washing and drying the obtained filter cake to obtain Form A.

[0082] In some embodiments, the raw materials of the preparation method only include the compound shown in Formula I, anhydrous ethanol, water and concentrated hydrochloric acid solution.

[0083] In some embodiments, the method for preparing the crystalline form A of the hydrochloride hydrate of the compound shown in Formula I further includes: cooling the solution of the crystalline form A of the hydrochloride hydrate of the compound shown in Formula I for crystallization to obtain a single crystal of the crystalline form A of the hydrochloride hydrate of the compound shown in Formula I, wherein the solvent of the solution of the crystalline form A of the hydrochloride hydrate of the compound shown in Formula I is a mixture of ethanol and water.

[0084] Wherein, the temperature of the mixture of ethanol and water may be 55-75°C, more preferably 60-70°C, further preferably 64-66°C.

[0085] Wherein, the volume ratio of ethanol to water in the mixture of ethanol and water may be 10:1 to 6:1, preferably 10:1 to 8:1, and more preferably 9:1.

[0086] Wherein, the cooling may be cooling to 20°C to 30°C.

[0087] The present invention also provides a hydrochloride hydrate of the compound of formula I, in which the molar ratio of the compound of formula I, HCl and water is 1:x:y, x is greater than 0 but not greater than 3, and y is greater than 0 but not greater than 3.

[0088] In some embodiments, in the hydrochloride hydrate, the molar ratio of the compound of formula I, HCl and water is 1:2:2.

[0089] The present invention also provides a pharmaceutical composition, comprising at least one of the hydrochloride hydrate of the compound represented by Formula I described in any of the above schemes and the crystalline form A of the hydrochloride hydrate of the compound represented by Formula I described in any of the above schemes, and at least one pharmaceutically acceptable carrier.

[0090] In some embodiments, the carrier is at least one of an excipient, a diluent, and a vehicle.

[0091] The present invention also provides use of the hydrochloride hydrate, crystal form A or pharmaceutical composition of the compound of formula I described in any of the above schemes in the preparation of a drug for preventing or treating Janus kinase-mediated disorders, diseases or conditions.

[0092] The present invention also provides a method for preventing or treating Janus kinase-mediated disorders, diseases or conditions in a subject in need thereof, comprising administering to the subject an effective amount of a hydrochloride hydrate, crystalline form A or pharmaceutical composition of the compound of formula I described in any of the above schemes.

[0093] The present invention also provides a hydrochloride hydrate, crystal form A or pharmaceutical composition of the compound of formula I described in any of the above schemes for preventing or treating Janus kinase-mediated disorders, diseases or conditions.

[0094] In some embodiments, the Janus kinase is at least one of JAK1, JAK2, JAK3 and TyK2.

[0095] In some embodiments, the Janus kinase-mediated disease is at least one of a hyperproliferative disease, cancer (e.g., leukemia and lymphoma), immune and inflammatory disorders (e.g., transplant rejection, asthma, chronic obstructive pulmonary disease, allergies, rheumatoid arthritis, psoriasis, atopic dermatitis, Crohn's disease, ulcerative colitis, amyotrophic lateral sclerosis, and multiple sclerosis).

[0096] In some embodiments, the Janus kinase-mediated disease is at least one of leukemia, lymphoma, transplant rejection, asthma, chronic obstructive pulmonary disease, allergy, rheumatoid arthritis, psoriasis, atopic dermatitis, Crohn's disease, ulcerative colitis, amyotrophic lateral sclerosis, and multiple sclerosis.

[0097] Without violating the common sense in the art, the above-mentioned preferred conditions can be arbitrarily combined to obtain the preferred embodiments of the present invention.

[0098] The reagents and raw materials used in the present invention are commercially available.

[0099] The positive and progressive effects of the present invention are that the salt form and the crystal form of the present invention have good JAK kinase inhibitory activity, solubility, stability and bioavailability, and enhance the developability of the oral preparation of the compound represented by Formula I.

[0100] Definition and Description

[0101] Unless otherwise specified, the following terms and phrases used herein are intended to have the following meanings. A particular term or phrase should not be considered to be uncertain or unclear in the absence of a special definition, but should be understood according to its ordinary meaning. When a trade name appears in this article, it is intended to refer to its corresponding commercial product or its active ingredient.

[0102] The term "pharmaceutically acceptable carrier" used in this article refers to pharmaceutical excipients, which refer to all substances contained in pharmaceutical preparations other than active ingredients, such as the pharmaceutical excipients specified in Part IV of the Pharmacopoeia of the People's Republic of China (2015 edition or 2020 edition).

[0103] The term "pharmaceutical composition" as used herein refers to a composition containing specified active ingredients and which can be prepared into the same dosage form.

[0104] The term "subject" as used herein refers to any animal that is about to or has been administered the compound or composition according to the embodiments of the present invention, preferably a mammal, and most preferably a human. As used herein, the term "mammal" includes any mammal. Examples of mammals include, but are not limited to, cattle, horses, sheep, pigs, cats, dogs, mice, rats, rabbits, guinea pigs, monkeys, humans, etc., with humans being the most preferred.

[0105] As used herein, the term "prophylactically or therapeutically effective amount" refers to an amount of a compound that is sufficient to effectively prevent or treat a disorder, disease or condition described herein when administered to a subject. The amount of a compound that constitutes a "prophylactically or therapeutically effective amount" will vary depending on the compound, the type of disorder, disease or condition and its severity, and the age of the subject, but can be adjusted as needed by those skilled in the art.

[0106] As used herein, the term "treat" refers to therapeutic therapy. When referring to a specific disease or condition, "treat" means at least one of the following: (1) alleviating one or more biological manifestations of the disorder, disease, or condition, (2) interfering with one or more biological manifestations of the disorder, disease, or condition, (3) improving or eliminating one or more symptoms, effects, or side effects associated with the disorder, disease, or condition, or one or more symptoms, effects, or side effects associated with the treatment of the disorder, disease, or condition; and (4) slowing the progression of one or more biological manifestations of the disorder, disease, or condition.

[0107] The present invention uses the following abbreviations:

[0108] XRPD X-ray powder diffraction

[0109] DSC Differential Scanning Calorimetry

[0110] TGA Thermogravimetric Analysis

[0111] rpm Revolutions per minute

[0112] ATP Adenosine triphosphate

[0113] DTT DL-dithiothreitol

[0114] SEB Supplier of Enzymatic Buffer, enzyme reaction buffer additive

[0115] AUClast Area under the plasma concentration-time curve

[0116] Cmax peak concentration

[0117] min

[0118] -RH Relative humidity

[0119] XRPD Characterization Method

[0120] Unless otherwise specified, the XRPD data described in the present invention (including but not limited to the attached Figure 1 , 4 -5, 8-12) were measured under the following conditions:

[0121] Sample preparation: Take an appropriate amount of the sample to be tested and evenly distribute it in the sample holder, and press it flat with a clean glass plate so that the surface of the sample is flush with the surface of the sample holder.

[0122] The instrument used is a Bruker D2 Phaser X-ray powder diffractometer, where the detector is a PSD LynxEye detector.

[0123] Diffractometer parameter settings: goniometer diameter, 282.2 mm; divergence slit, 1.0 mm; primary cable slit, 2.5 degrees; secondary cable slit, 2.5 degrees; anti-air scattering assembly, 1.0 mm; light pipe element, copper; light pipe parameters, voltage 30 kV, current 10 mA;

[0124] Scan parameter settings: scan type, Locked coupled; scan mode, Continuous PSD fast mode; rotation speed, 20 degrees / minute; scan range, 3 degrees to 40 degrees (2θ); scan step, 0.02 degrees (2θ); scan speed, 0.2 seconds / step; detector opening, 4.5 degrees.

[0125] DSC Characterization Method

[0126] Unless otherwise specified, the DSC data described in the present invention are measured under the following conditions:

[0127] Instrument: Discovery DSC250 differential scanning calorimeter, sample plate type is punch hole;

[0128] The heating range is 25℃-300℃, and the heating rate is 10℃ / min;

[0129] TGA Characterization Method

[0130] Unless otherwise specified, the TGA data described in the present invention are measured under the following conditions:

[0131] Instrument: Discovery 55 Thermogravimetric Analyzer

[0132] Sample pan type: default open aluminum pan;

[0133] Heating range: 10℃-300℃, heating rate: 10℃ / min. BRIEF DESCRIPTION OF THE DRAWINGS

[0134] Figure 1 is the XRPD pattern of Form A;

[0135] Figure 2 is the DSC spectrum of Form A;

[0136] Figure 3 is the TGA spectrum of Form A;

[0137] Figure 4 is the XRPD pattern of Form C;

[0138] Figure 5 is the XRPD pattern of Form D;

[0139] Figure 6 is the XRPD pattern of Form E;

[0140] Figure 7 is the XRPD pattern of Form F;

[0141] Figure 8 This is the XRPD pattern of Form A after being placed in a stabilization chamber at 40°C / 60%-RH for 30 days;

[0142] Fig. 9 This is the XRPD pattern of Form A after being placed in a stabilization chamber at 25°C / 75%-RH for 30 days;

[0143] Fig.10 This is the XRPD spectrum of the solid precipitated after adding Form A to the saturated solution system in Example 5;

[0144] Fig.11 This is the XRPD spectrum of the solid precipitated after adding Form E to the saturated solution system in Example 5;

[0145] Fig.12 This is the XRPD spectrum of the solid precipitated after adding Form F to the saturated solution system in Example 5;

[0146] Fig.13 This is the single crystal shape of Form A obtained in Example 8;

[0147] Fig.14 This is the XRPD pattern of the Form A single crystal obtained in Example 8;

[0148] Fig.15 Schematic diagram of the asymmetric structural unit of the Form A single crystal obtained in Example 8.

[0149] Figure 1 , 4 -5, 8-12, 14, the unit of the vertical axis intensity is counts, and the unit of the horizontal axis 2θ angle is degree (°). Figure 6-7 The unit of the horizontal coordinate 2θ angle is degree (°). DETAILED DESCRIPTION

[0150] The present invention is further described below by way of examples, but the present invention is not limited to the scope of the examples. The experimental methods in the following examples without specifying specific conditions are carried out according to conventional methods and conditions, or selected according to the product specifications.

[0151] In the following examples, unless otherwise specified, mass spectra were detected using a Waters Acquity Xevo G2-XS QTofUPLC / MS ultra-high performance liquid chromatography high-resolution mass spectrometry system. 1 H-NMR was detected by Bruker AVANCE III 400 MHz nuclear magnetic resonance instrument or Bruker AVANCE III HD 300 MHz nuclear magnetic resonance instrument, and HPLC was detected by Agilent 1260 high performance liquid chromatograph.

[0152] If not otherwise specified, the concentrated hydrochloric acid solution described in the present invention refers to an aqueous solution of hydrogen chloride (HCl), and its concentration is not less than 8 mol / L.

[0153] Unless otherwise specified, room temperature in the present invention refers to 20-30°C.

[0154] Example 1. Synthesis of the compound represented by formula I

[0155] Synthesis of compound 4

[0156]

[0157] SM1-1 (54.60 g, 175.0 mmol) and dichloromethane (1 L) were added to a 3 L round-bottom flask. After cooling to 0 ° C, trifluoroacetic acid (200 mL) was slowly added dropwise. After the addition, the mixture was stirred at room temperature for 10 hours. HPLC showed that the reaction was complete, and the trifluoroacetic acid was removed in vacuo, and then diluted with dichloromethane (500 mL). The resulting solution was washed with a NaHCO3 aqueous solution, dried and concentrated, and purified by silica gel column chromatography to obtain white solid compound 4 (23.00 g, yield 62.0%).

[0158] NMR data of compound 4: 1 H NMR (400 MHz, DMSO-d 6 ) δppm: 7.24 (d, J=8.4Hz, 1H), 7.05 (dd, J=1.1Hz, 6.8Hz, 1H), 6.97-7.01 (m, 2H), 5.31 (br, 2H).

[0159] Synthesis of compound 5

[0160]

[0161] Compound 4 (23.00 g, 108.5 mmol), triethylamine (39.46 g, 390.0 mmol) and acetonitrile (400 mL) were added to a 2 L round bottom flask. After cooling to 0 ° C, cyclopropyl chloride (40.23 g, 384.9 mmol) was slowly added dropwise. After the addition, the mixture was stirred at room temperature for 8 hours. HPLC showed that the reaction was complete. The resulting reaction solution was concentrated to dryness, then washed with water, and filtered to obtain a solid. The resulting solid was subjected to column chromatography to obtain a white solid compound 5 (29.46 g, yield 97.0%).

[0162] NMR data of compound 5: 1 H NMR (400 MHz, DMSO-d 6 )δppm: 11.14 (br, 1H), 8.05 (s, 1H), 7.50-7.52 (m, 1H), 7.19-7.28 (m, 2H), 1.92-1.99 (m, 1H), 0.82-0.84 (m, 4H).

[0163] Synthesis of the compound shown in formula I

[0164]

[0165] Compound 5 (29.46 g, 105.2 mmol), SM2-1 (41.88 g, 113.4 mmol), K2CO3 (26.12 g, 189.0 mmol), Pd(dppf)Cl2.CH2Cl2 (6.91 g, 8.5 mmol), H2O (120 mL) and 1,4-dioxane (600 mL) were added to a 2 L round bottom flask. The mixture was heated to reflux for 10 hours under N2 protection. HPLC showed that the reaction was complete, ice water was added to quench the reaction, and a solid was filtered to obtain the solid. The obtained solid was subjected to column chromatography to obtain the compound shown in Formula I (27.42 g, yield 58.9%) as a white solid.

[0166] Characterization data of the compound shown in formula I: 1 H (400 Hz, DMSO-d 6 ) δppm: 8.08 (1H, dd, J = 9.2, 7.3Hz), 8.03 (1H, dd, J = 9.3, 1.6Hz), 7.84-7.78 (3H, m), 7.60 (1H, dd, J = 7. 1, 1.6Hz), 4.66 (2H, s), 3.90-3.89, (4H, m), 3.70-3.68 (4H, m), 1.97-1.94 (1H, m), 1.08-1.04 (4H, m). LC / MSm / z: 443.2(M+H).

[0167] Example 2. Preparation of Form A of the Hydrochloride Hydrate of the Compound of Formula I

[0168] Add the compound of formula I (1.5 g, 3.4 mmol) to a round-bottom flask, then add 13.5 mL of anhydrous ethanol and 1.5 mL of purified water, and stir the resulting mixture to 50°C. Then add 0.62 mL of concentrated hydrochloric acid solution (12 mol / L, 7.4 mmol) dropwise. After the addition is complete, keep the hydrochloride solution of the compound of formula I at 50°C and stir for 1 hour, then cool naturally to 25°C at room temperature, and then keep at 25°C and stir for 48 hours to precipitate crystals, and stop stirring. Filter, wash the resulting filter cake with a small amount of ethanol, collect, and vacuum dry at 40°C for 6 hours. The resulting crystals are Form A.

[0169] The XRPD pattern of Form A is as follows: Figure 1 The X-ray powder diffraction peaks expressed in 2θ angles are shown in Table 1.

[0170] The DSC results of Form A are as follows: Figure 2 As shown, DSC shows the first endothermic peak at 165.14°C, with a thermal enthalpy of 180.63 J / g and an onset temperature of 140.82°C; the other endothermic peak is at 197.70°C, with a thermal enthalpy of 23.689 J / g and an onset temperature of 179.20°C.

[0171] The TGA results of Form A are as follows: Figure 3 As shown, the sample has a weight loss of 8.3143% before 148.00°C and a weight loss of 4.7637% at 148-228°C.

[0172] The chloride ion content was determined using a Mettler-Toledo T5 potentiometric titrator, and the results showed that the chloride ion content in the sample was 12.3%, which was basically consistent with the sample containing two chloride ions (the theoretical value of chloride ions in the sample containing two hydrochlorides was 12.9%). The moisture content was determined using a Mettler-Toledo KF titrator V30S, and the results showed that the water content in the sample was 8.0%, which was basically consistent with the sample containing two crystal waters (the theoretical value of moisture in the sample containing two crystal waters was 6.5%). It can be determined that the crystalline form A is a hydrochloride hydrate of the compound of formula I, in which the molar ratio of the compound of formula I, HCl and water is 1:2:2.

[0173] Comparative Example 1. Attempt to prepare Form A using other solvent systems

[0174] (1) Crystallization in THF / water system

[0175] Add the compound of formula I (1.0 g, 2.3 mmol) to a round-bottom flask, then add 9 mL of THF and 1 mL of pure water, and stir the resulting mixture to 60°C. Then drop 0.41 mL of concentrated hydrochloric acid solution (12 mol / L, 4.9 mmol). After the addition is complete, keep the hydrochloride solution of the compound of formula I at 60°C and stir for 1 hour, then cool naturally to 25°C at room temperature, then keep at 25°C and stir for 48 hours to precipitate crystals, and stop stirring. Filter, wash the resulting filter cake with a small amount of THF and dry, and the resulting solid is Form C, and its XRPD spectrum is as follows: Figure 4 As shown, it can be seen that the obtained crystal form C is different from the crystal form A.

[0176] (2) Crystallization in DCM / MeOH system

[0177] Add the compound of formula I (0.5 g, 1.1 mmol) to a round-bottom flask, then add 10 mL of DCM and 10 mL of MeOH, and stir the resulting mixture to 40°C. Then add 4M hydrochloric acid solution (0.6 mL, 2.4 mmol) dropwise. After the addition is complete, keep the hydrochloride solution of the compound of formula I at 40°C and stir for 1 hour, then cool naturally to 25°C at room temperature, then keep at 25°C and stir for 48 hours to precipitate crystals, and stop stirring. Filter, wash the resulting filter cake with a small amount of DCM and dry, and the resulting solid is Form D, and its XRPD spectrum is as follows: Figure 5 As shown. It can be seen that the obtained crystal form D is different from the crystal form A.

[0178] Comparative Example 2. Preparation of Hydrochloride Form E of the Compound of Formula I

[0179] 10 mg of the hydrochloride hydrate of the compound of formula I, Form A, was added to 0.3 mL of acetonitrile, stirred at room temperature for 3 days, and filtered to obtain Form E. The XRPD spectrum of the crystal form is as follows Figure 6 shown.

[0180] The present invention is attached Figure 6 The detection instrument used for the XRPD spectrum shown is a Bruker D8 Advance X-ray diffractometer, wherein the X-ray tube parameters are set as follows: X-ray wavelength, Cu: Voltage, 40 kV; current, 40 mA; sample rotation speed: 15 rpm; scanning range: 3 to 40 degrees (2θ); scanning speed: 10 degrees / min.

[0181] Comparative Example 3. Preparation of Hydrochloride Form F of the Compound of Formula I

[0182] 10 mg of the hydrochloride hydrate of the compound of formula I, Form A, was added to a 4 mL vial, added to 0.5 mL of DMSO, and stirred to dissolve. Another 40 mL round-bottom flask was taken, and 4 mL of acetonitrile was added. The vial was placed in the round-bottom flask, which was covered and left at room temperature to slowly crystallize. After 3 days, Form F was obtained by filtration. Its XRPD is as follows: Figure 7 shown.

[0183] The present invention is attached Figure 7 The detection instrument used for the XRPD spectrum shown is a Bruker D8 Advance X-ray diffractometer, wherein the X-ray tube parameters are set as follows: X-ray wavelength, Cu: Voltage, 40 kV; current, 40 mA; sample rotation speed: 15 rpm; scanning range: 3 to 40 degrees (2θ); scanning speed: 10 degrees / min.

[0184] Example 3. JAK kinase inhibitory activity test of the hydrochloride hydrate of the compound of formula I in form A

[0185] Cisbio The in vitro enzyme activity biochemical assay of the compound was performed using a HTRF kinEASE-TK kit (catalog number: 62TKOPEC). The substrate, kinase reaction buffer, detection buffer, allophycocyanin modifier (XL-665) labeled streptavidin, europium (EU) labeled specific phosphorylation antibody and SEB used in the assay were all provided by the kit. JAK1 used in the assay was purchased from Invitrogen (catalog number PV4774), JAK2, JAK3 and TYK2 were purchased from Carna Biosciences, Inc. (catalog numbers: 08-045, 08-046, 08-147, respectively), and DTT was purchased from Sigma (catalog number 43816).

[0186] The compound (the hydrochloride hydrate crystal form A of the compound shown in Formula I prepared according to the method of Example 2 or Tofacitinib citrate (Shanghai Haoyuan Chemical Technology Co., Ltd., HY-40354A)) was diluted 3 times in DMSO for 10 times, and the concentration of the obtained compound dilution was 100 times the final test concentration. Then, the concentration of the compound dilution was further diluted to 2.5 times the final test concentration with kinase reaction buffer to obtain a compound test solution.

[0187] The enzymatic reaction was performed in a white 384-well polypropylene plate (Greniner, catalog number: 784075) with a total reaction volume of 10 uL containing 500 ng / mL JAK1, 6 ng / mL JAK2, 37 ng / mL JAK3, 100 ng / mL TYK2, 1 uM substrate and 1 mM ATP (Sigma-Aldrich, catalog number: A7699).

[0188] 4uL of the compound to be tested solution was added to the wells of a 384-well polypropylene plate, followed by the addition of 2uL of JAK1 (additional buffer additives MgCl2, MnCl2 and DTT were added to final concentrations of 5mM, 1mM and 1mM, respectively), JAK2 (additional buffer additives MgCl2 and DTT were added to final concentrations of 5mM and 1mM, respectively), JAK3 (additional buffer additives MgCl2 and DTT were added to final concentrations of 5mM and 1mM, respectively), TYK2 (additional buffer additives MgCl2, MnCl2, DTT and SEB were added to final concentrations of 5mM, 1mM, 1mM and 12.5nM, respectively) diluted in kinase reaction buffer and incubated at room temperature for 15 minutes for pretreatment. Enzymatic reactions were initiated by adding a mixture of 2uL substrate and 2uL ATP prepared in kinase reaction buffer. After 30 minutes of reaction at room temperature, 5uL of streptavidin labeled with allophycocyanin modifier (XL-665) prepared in detection buffer and 5uL of europium-labeled specific phosphorylated antibody prepared in detection buffer were added to stop the enzymatic reaction and generate signals. After incubation for 1 hour at room temperature, the plate was read in a Molecular Devices SpectraMAX Paradigm multifunctional microplate reader with the following settings: excitation is 340nm / emission 1 is 616nm / emission 2 is 665nm. The ratio (Ratio) of the emission light signal of the acceptor and donor of each well was calculated as follows: Ratio = signal 665nm / signal 616nm*10000, wherein signal 665nm is the signal of 665nm and signal 616nm is the signal of 616nm. The percentage of inhibition was calculated as follows: percentage of inhibition = 100% - the percentage of the ratio value of the treated compound / the percentage of the ratio value of the treated DMSO vehicle (DMSO vehicle is a blank control group without drugs).

[0189] Dose-response curves were generated and IC was calculated by nonlinear sigmoidal curve fitting using GraphPad Prism software. 50 The results are as follows:

[0190] Table 2. IC of Form A of the hydrochloride hydrate of the compound of Formula I 50data

[0191]

[0192] Example 4. Solubility test of the compound represented by formula I and crystal form A

[0193] Weigh the samples ground into fine powder and add them to a certain volume of solvent (ambient temperature: 25°C ± 5°C), shake vigorously for 30 seconds every 5 minutes, and observe the dissolution within 30 minutes. If there are no visible solute particles or droplets, it is considered to be completely dissolved. According to this method, the dissolution of the hydrochloride hydrate of the compound represented by Formula I and the free base form of the compound represented by Formula I in common solvents was observed, and the results are shown in Table 3. Among them, "readily soluble" means that 1g (mL) of solute can be dissolved in a solvent with a volume of 1mL≤V<10mL; "soluble" means that 1g (mL) of solute can be dissolved in a solvent with a volume of 10mL≤V<30mL; "slightly soluble" means that 1g (mL) of solute can be dissolved in a solvent with a volume of 30mL≤V<100mL; "slightly soluble" means that 1g (mL) of solute can be dissolved in a solvent with a volume of 100mL≤V<1000mL; "very slightly soluble" means that 1g (mL) of solute can be dissolved in a solvent with a volume of 1000mL≤V<10000mL; "almost insoluble" or "insoluble" means that 1g (mL) of solute cannot be completely dissolved in a solvent with a volume of 10000mL.

[0194] Table 3. Solubility of the free base of the compound of formula I and its hydrochloride hydrate in Form A

[0195] Solvent Free base of the compound shown in formula I Crystalline Form A Methanol Slightly soluble Soluble Ethanol Slightly soluble Dissolve water Very slightly soluble Dissolve Simulated gastric fluid Insoluble Slightly soluble

[0196] Example 5. Stability test of different crystal forms

[0197] Test whether temperature and humidity have an effect on the stability of the hydrochloride hydrate of the compound represented by Formula I in Form A:

[0198] The crystal form A was placed in a stabilization chamber, and the temperature and humidity were controlled at 40°C / 60%-RH and 25°C / 75%-RH, respectively. After 30 days, the obtained solid was compared with its initial crystal form A: its appearance did not change significantly, the content of the compound represented by Formula I measured by HPLC (as shown in Table 4) did not change significantly, and the XRPD (XRPD patterns obtained after treatment at 40°C / 60%-RH and 25°C / 75%-RH, respectively, were shown in Table 4) did not change significantly. Figure 8 and Fig. 9 ) also showed no obvious change, indicating that Form A has good stability.

[0199] Table 4. Content of the compound represented by formula I before and after crystal form A is placed

[0200] sample The content of the compound shown in formula I Initial Crystal A 99.901％ Crystal A was placed at 40℃ / 75%-RH for 30 days. 99.905％ Crystal A was placed at 25℃ / 75%-RH for 30 days. 99.899％

[0201] Comparison of thermodynamic stability of Form A, Form E and Form F:

[0202] Crystalline Form A, Crystalline Form E and Crystalline Form F were dissolved in a mixed solvent of ethanol / water (volume ratio = 9:1) to prepare saturated solutions. The solution was heated to 50°C and 10 mg of Crystalline Form A, Crystalline Form E and Crystalline Form F were added under stirring. The mixture was stirred at 50°C for 24 hours and then filtered. The XRPD patterns of the crystals obtained were measured (the XRPD patterns of the crystals precipitated by adding Crystalline Form A, E and F are shown in the figure below). Fig.10 , 11 and 12). The obtained XRPD pattern and Figure 1 Comparative analysis of the XRPD spectrum of Form A shown in the figure shows that the crystals precipitated from the system with Form A added are still Form A, while Form E and Form F are both converted into Form A, indicating that Form A has better thermodynamic stability than Form E and Form F.

[0203] Example 6. Single-dose PK study

[0204] Two groups of SD rats (3 females in each group) were administered a single oral dose of 3 mg / kg of the free base of the compound of Formula I and a single oral dose of 3 mg / kg of the hydrochloride hydrate of the compound of Formula I in Form A. The drug was administered in the form of a 0.5% CMC-Na suspension. Samples were taken at different times until 24 hours, and the drug concentration of the compound of Formula I in plasma was analyzed by LC-MS / MS, and the final calculated results are shown in Table 8.

[0205] Chromatographic conditions of the LC-MS / MS method for the compound shown in Formula I:

[0206] Instrument model, SHIMADZU LC-30AD liquid phase system; Chromatographic column, ACQUITY BEH C18 (2.1×50 mm, 1.7 μm); column temperature, 40°C; injection volume, 1 μL; flow rate, 0.6 mL / min; running time, 6 min; mobile phase gradient settings are shown in Table 5.

[0207] Table 5.

[0208]

[0209] Mass spectrometry conditions:

[0210] Instrument model, AB SCIEX TRIPLE QUAD 6500 mass spectrometer; ion source, electrospray ionization (ESI); ionization mode, positive ion scan; multiple ion (MRM) ion pairs are shown in Table 6; instrument parameters are shown in Table 7.

[0211] Table 6.

[0212]

[0213]

[0214] *: Internal standard compound purchased from MedChemExpress (Catalog No.: HY-18300), same as in Table 7.

[0215] Table 7.

[0216] parameter Compounds shown in formula I GLPG0634 Spray voltage (v) 5000 5000 Ion source temperature (℃) 550 550 Collision gas (psi) 10 10 Curtain gas (psi) 40 40 Atomizing gas (psi) 50 50 Auxiliary gas (psi) 40 40 Declustering voltage (v) 110 110 Inlet voltage (V) 12 12 Collision voltage (v) 47 39 Collision cell exit voltage (v) 10 10

[0217] Table 8. PK data of the hydrochloride hydrate of the compound of formula I, Form A, and the free base of the compound of formula I

[0218] Solid form AUClast Cmax Free base 987h*ng / mL 352 ng / mL Crystalline Form A 1390h*ng / mL 456 ng / mL

[0219] Example 7. Study on the Hygroscopicity of Form A, Form C, Form D, Form E and Form F

[0220] About 500 mg of each of Form A, Form C, Form D, Form E and Form F were taken, accurately weighed and placed in a constant humidity sealed container, left at 25°C / 75%-RH for 5 days, and then taken out to measure their moisture absorption and weight gain. The results are shown in Table 9, indicating that Form A has the lowest hygroscopicity.

[0221] Table 9: Hygroscopicity data of each crystal form

[0222] Crystal form Crystalline Form A Crystalline Form C Crystal form D Crystal Form E Form F Moisture absorption and weight gain 2.4％ 6.1％ 8.1％ 5.3％ 4.9％

[0223] Example 8. Single crystal testing and structural analysis of Form A

[0224] Weigh 35 mg of the crystalline form A sample of the hydrochloride hydrate of the compound of formula I prepared in Example 2, dissolve it in 1 mL of ethanol / water (9:1, v / v) at 65°C, and cool it to room temperature for crystallization to obtain long rod-shaped crystals ( Fig.13 Then some crystals were selected for XRPD diffraction study ( Fig.14 ), Fig.14 There are characteristic diffraction peaks at the following 2θ angles: 5.9°±0.2°, 7.4°±0.2°, 11.6°±0.2°, 21.7°±0.2° and 23.8°±0.2°, which are consistent with Figure 1 The results of single crystal structure analysis show that Form A belongs to the monoclinic system, P21 / c space group, and its unit cell parameters are α=90°, β=101.262(3)°, γ=90°, Fig.15Schematic diagram of the asymmetric structural unit of the crystal, wherein the asymmetric unit contains one cation of the compound represented by Formula I, two chloride ions and two crystal waters.

[0225] Instruments, software and methods used in the experiment:

[0226] X-ray powder diffraction (XRPD)

[0227] The solid samples obtained in the experiment were analyzed by X-ray powder diffractometer Bruker D8 Advance (Bruker, GER). The 2θ scanning angle was from 3° to 45°, the scanning step was 0.02°, and the exposure time was 0.12 s. When testing the samples, the tube voltage and current were 40 kV and 40 mA, respectively, and the sample pan was a zero background sample pan.

[0228] Single Crystal X-ray Diffraction (SCXRD)

[0229] A single crystal with suitable morphology and size was selected from the cultured single crystal samples, and the single crystal was glued to the loop ring, and then the single crystal sample was placed on the crystal stage. The single crystal sample pre-experiment and single crystal diffraction data collection were performed using a SuperNova (Rigaku, JPN) single crystal diffractometer (Cu target light source, ) was carried out at 253.0K, and the diffraction data were analyzed and processed using the CrysAlisPro software package.

[0230] The angle range of the diffraction data collection at 253.0K is 4.4340°<θ<73.8890°, and 8128 diffraction points are collected from this angle range. The unit cell parameters and orientation matrix of the crystal are obtained by analyzing them using the CrysAlisPro program and refining them by the least square method. The completeness of the data collection corresponding to the highest θ angle (θ=66.97°) is 99.94%.

[0231] Data Restoration

[0232] The CrysAlisPro 1.171.39.46 (Rigaku Oxford Diffraction, 2018) program was used to restore and integrate each frame of the diffraction image collected by the detector, and a total of 16159 diffraction points were collected, including 4275 independent diffraction points. The SCALE3 ABSPACK scaling algorithm was used to correct the diffraction data for absorption. The linear absorption coefficient of X-rays is 3.404 mm -1, the minimum transmittance (Tmin) is 0.586, the maximum transmittance (Tmax) is 1.000. The intensities of all equivalent diffraction points are basically equal within the experimental error range, and its Rint is 4.30%.

[0233] Structural analysis and refinement

[0234] The single crystal structure was analyzed using OLEX2 software, and the XS (Sheldrick, 2008) program was used to perform a preliminary structural analysis on the diffraction data and determine that the space group to which the crystal belongs is P21 / c. The XH (Sheldrick, 2008) program was then used for structural refinement. The coordinates of all non-hydrogen atoms were determined by several rounds of difference Fourier synthesis, and then all non-hydrogen atoms were anisotropically refined using the full matrix least squares method. All hydrogen atoms were calculated using theoretical hydrogenation.

[0235] Crystal structure diagram

[0236] The crystal structure diagram and atomic thermal vibration ellipsoid diagram were drawn using Diamond software.

[0237] Although the specific embodiments of the present invention are described above, it should be understood by those skilled in the art that this is only for illustration and the protection scope of the present invention is defined by the appended claims. Those skilled in the art may make various changes or modifications to these embodiments without departing from the principles and essence of the present invention, but these changes and modifications all fall within the protection scope of the present invention.

Claims

1. Form A of the hydrochloride hydrate of the compound represented by formula I, characterized in that: The crystal form A is a single crystal, which belongs to the monoclinic system, the P21 / c space group, and the unit cell parameters are α=90°,β=101.262(3)°,γ=90°, the unit cell volume is The X-ray powder diffraction pattern has characteristic diffraction peaks at the following 2θ angles: 5.9°±0.2°, 7.4°±0.2°, 11.6°±0.2°, 21.7°±0.2° and 23.8°±0.2°; 2. The crystalline form A of the hydrochloride hydrate of the compound of formula I as claimed in claim 1, characterized in that: The X-ray powder diffraction pattern of the crystalline form A has characteristic diffraction peaks at the following 2θ angles: 5.9°±0.2°, 7.4°±0.2°, 11.6°±0.2°, 17.7°±0.2°, 17.8°±0.2°, 21.7°±0.2°, 23.6°±0.2°, 23.8°±0.2°, 29.0°±0.2°, 30.4°±0.2° and 34.9°±0.2°.

3. The crystalline form A of the hydrochloride hydrate of the compound of formula I according to claim 1, characterized in that: The X-ray powder diffraction pattern of the crystalline form A has characteristic diffraction peaks at the following 2θ angles: 5.9°±0.2°, 7.4°±0.2°, 8.9°±0.2°, 11.6°±0.2°, 14.6°±0.2°, 17.7°±0.2°, 17.8°±0.2°, 18.7°±0.2°, 19.5°±0.2°, 21.0°±0.2°, 21.7°±0.2°, 21.9°±0.2°, 22.6°±0.2°, 23.6°±0.2°, 23.8°±0.2°, 29.0°±0.2°, 30.4°±0.2°, 33.3°±0.2°, 34.9°±0.2° and 37.7°±0.2°.

4. The crystalline form A of the hydrochloride hydrate of the compound of formula I as claimed in claim 3, characterized in that: The X-ray powder diffraction pattern of the crystalline form A has characteristic diffraction peaks at the following 2θ angles: 5.9°±0.2°, 6.6°±0.2°, 7.4°±0.2°, 8.9°±0.2°, 10.5°±0.2°, 11.6°±0.2°, 12.3°±0.2°, 13.9°±0.2°, 14.2°±0.2°, 14.6°±0.2°, 17.1°±0.2°, 17.7°±0.2°, 17.8°±0.2°, 18.7°±0.2°, 19.5°±0.2°, 20.4°±0.2°, 21.0°±0.2°, 21.2°±0.2°, 21.7°±0.2°, 21.8°±0.2°, 21.9°±0.2°, 22.1°±0.2°, 22.2°±0.2°, 22.3°±0.2°, .9°±0.2°, 22.6°±0.2°, 23.6°±0.2°, 23.8°±0.2°, 24.9°±0.2°, 26.6°±0.2°, 26.8°±0.2°, 28.3°±0.2°, 29.0°±0.2°, 30.4°±0.2°, 31.2°±0.2°, 32.3°±0.2°, 33.3°±0.2°, 34.2°±0.2°, 34.9°±0.2°, 35.6°±0.2°, 35.9°±0.2°, 36.4°±0.2°, 36.9°±0.2°, 37.7°±0.2°, 39.3°±0.2° and 39.8°±0.2°.

5. The crystalline form A of the hydrochloride hydrate of the compound of formula I as claimed in claim 3, characterized in that: The X-ray powder diffraction pattern of the crystalline form A has characteristic diffraction peaks at the following 2θ angles: 5.9°, 6.6°, 7.4°, 8.9°, 10.5°, 11.6°, 12.3°, 13.9°, 14.2°, 14.6°, 17.1°, 17.7°, 17.8°, 18.7°, 19.5°, 20.4°, 21.0°, 21.2°, 2 1.7°, 21.9°, 22.6°, 23.6°, 23.8°, 24.9°, 26.6°, 26.8°, 28.3°, 29.0°, 30.4°, 31.2°, 32.3°, 33.3°, 34.2°, 34.9°, 35.6°, 35.9°, 36.4°, 36.9°, 37.7°, 39.3° and 39.8°.

6. The crystalline form A of the hydrochloride hydrate of the compound of formula I according to claim 1, characterized in that: The XRPD pattern analysis data of the crystal form A is shown in the following table: 。 7. The crystalline form A of the hydrochloride hydrate of the compound of formula I as described in any one of claims 1 to 6, characterized in that: The thermogravimetric analysis curve of the crystal form A shows a weight loss of 8.31±0.50% before 148°C, and a weight loss of 4.76±0.50% between 148°C and 228°C.

8. The crystalline form A of the hydrochloride hydrate of the compound of formula I as claimed in claim 7, characterized in that: The thermogravimetric analysis curve of the crystal form A shows a weight loss of 8.31% before 148°C, and a weight loss of 4.76% between 148°C and 228°C.

9. The crystalline form A of the hydrochloride hydrate of the compound of formula I as claimed in claim 7, characterized in that: The thermogravimetric analysis curve of the crystal form A shows a weight loss of 8.3143% before 148°C, and a weight loss of 4.7637% between 148°C and 228°C.

10. The crystalline form A of the hydrochloride hydrate of the compound of formula I as described in any one of claims 1 to 6, characterized in that: The differential scanning calorimetry diagram of the crystal form A has absorption peaks at 165°C±3°C and 198°C±3°C, respectively; And / or, the differential scanning calorimetry diagram of the crystalline form A has absorption peaks with onset temperatures of 141°C±3°C and 179°C±3°C, respectively.

11. The crystalline form A of the hydrochloride hydrate of the compound of formula I according to claim 10, characterized in that: The differential scanning calorimetry diagram of the crystal form A has absorption peaks at 165.14° C. and 197.70° C. respectively; And / or, the differential scanning calorimetry diagram of the crystalline form A has absorption peaks with onset temperatures of 140.82° C. and 179.20° C., respectively.

12. The crystalline form A of the hydrochloride hydrate of the compound of formula I as described in any one of claims 1 to 6, characterized in that: In the crystalline form A, the molar ratio of the compound represented by formula I, hydrochloric acid and water is 1:2:

2.

13. A method for preparing the crystalline form A of the hydrochloride hydrate of the compound represented by formula I according to any one of claims 1 to 12, characterized in that: The following steps are involved: The compound of formula I, an organic solvent and water are mixed, the obtained mixture is stirred and heated to 50°C, and a concentrated hydrochloric acid solution is added to obtain a hydrochloride solution of the compound of formula I; wherein the organic solvent is anhydrous ethanol; the concentration of the concentrated hydrochloric acid solution is 12 mol / L; the ratio of the organic solvent, water, concentrated hydrochloric acid solution and the compound of formula I is 9 mL: 1 mL: 0.41 mL: 1 g; The hydrochloride solution of the compound of formula I is kept at 50° C. and stirred for 1 hour, then cooled to 25° C. and kept at 25° C. and stirred for 48 hours to precipitate crystals; a single crystal of the hydrochloride hydrate of the compound of formula I in form A is obtained.

14. A pharmaceutical composition comprising the crystalline form A of the hydrochloride hydrate of the compound of formula I according to any one of claims 1 to 12, and at least one pharmaceutically acceptable carrier.

15. The pharmaceutical composition according to claim 14, characterized in that The carrier is an excipient.

16. The pharmaceutical composition according to claim 14, characterized in that The carrier is a diluent.

17. The pharmaceutical composition according to claim 14, characterized in that The carrier is a solvent.

18. Use of the hydrochloride hydrate form A of the compound of formula I according to any one of claims 1 to 12 or the pharmaceutical composition according to any one of claims 14 to 17 in the preparation of a medicament for treating a disease mediated by Janus kinase; the disease mediated by Janus kinase is rheumatoid arthritis.

19. The use according to claim 18, characterized in that The Janus kinase is at least one of JAK1, JAK2, JAK3 and TyK2.

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