A salt of a ROCK inhibitor, a crystal form of the salt, a composition and a pharmaceutical use

Abstract

The present invention discloses a salt of a ROCK inhibitor, a crystal form of the salt, a composition and a pharmaceutical use. The salt and the crystal form of the salt are an acid addition salt of Compound I and any one of the following acids and its crystal form: hydrochloric acid, sulfuric acid, p-toluenesulfonic acid, benzenesulfonic acid, maleic acid, tartaric acid, oxalic acid, ethanesulfonic acid, 2-hydroxyethanesulfonic acid, camphorsulfonic acid, 1,5-naphthalenedisulfonic acid. The free base crystal form, the salt and the crystal form of the salt of Compound I provided by the present invention have stable physical and chemical properties and good biological properties, which are more conducive to the quality control and drug-likeness of drugs.

Description

[0001] This application claims the right of priority to the following two prior applications: a prior application filed with the China National Intellectual Property Administration on January 13, 2022, with the patent application number 202210039840.3 and the invention title "A Salt of a ROCK Inhibitor, Crystal Forms of the Salt, Compositions, and Pharmaceutical Uses"; and a prior application filed with the China National Intellectual Property Administration on January 5, 2023, with the patent application number 202310016976.7 and the invention title "A Salt of a ROCK Inhibitor, Crystal Forms of the Salt, Compositions, and Pharmaceutical Uses". The entire text of the prior applications is incorporated into this application by reference. Technical Field

[0002] The present invention relates to the field of medicinal chemistry, and particularly to a salt of a ROCK inhibitor, crystal forms of the salt, compositions, and pharmaceutical uses. Background Art

[0003] Idiopathic interstitial pulmonary fibrosis (IPF) is a chronic, diffuse interstitial lung disease of unknown cause with changes characteristic of usual interstitial pneumonia. The histological and / or high-resolution chest CT (HRCT) features of the lung in IPF patients are characteristic of usual interstitial pneumonia. IPF patients have an insidious onset, and the main clinical manifestations are dry cough and progressive dyspnea, which are obvious after activity. Due to its complex pathogenesis, the disease progresses irreversibly, and early diagnosis is difficult; after diagnosis, the survival rate of patients decreases significantly over time. Its 3-year survival rate is 50%, and the 5-year survival rate is only 20%, which is lower than that of most cancers (such as leukemia, breast cancer, colon cancer, uterine cancer, kidney cancer, etc.), and it is known as the "cancer that is not cancer". Currently, there are no definitely effective therapeutic drugs and methods for IPF. According to the "Chinese Expert Consensus on the Diagnosis and Treatment of Idiopathic Pulmonary Fibrosis" released in 2016, only 3 drugs are "conditionally recommended for use", namely pirfenidone, nintedanib, and antacid drugs. However, there is currently insufficient evidence to confirm that antacid drug treatment can delay the decline of lung function in IPF. Currently, the only drugs on the market for the IPF indication globally are pirfenidone and nintedanib; however, pirfenidone and nintedanib still have certain deficiencies in efficacy and safety, and the treatment options are limited. There is an urgent need to develop drugs with other target mechanisms to meet the urgent needs of clinical patients and the possible subsequent demands for combination drug use.

[0004] ROCK (Rho-associated protein kinase), also known as Rho kinase (Rho-associated kinase), belongs to serine / threonine protein kinases with a molecular mass of approximately 160 kD. It is the most well-studied Rho downstream target effector molecule to date. ROCK is distributed in tissues throughout the body, including ROCK1 (ROKβ, p160-ROCK) and ROCK2 (ROKα) subtypes, which regulate the assembly of actin filaments and actomyosin contraction, control the damage responses of various cells, especially epithelial cells, endothelial cells, and fibroblasts, and mediate the fibrotic process in the pathogenesis of IPF; ROCK also plays a role in many signaling pathways related to autoimmunity and inflammation. ROCK signaling can interfere with various fibrotic initiation conditions, reduce fibroblast activation and collagen deposition, and improve organ function. Preclinical studies have shown that both ROCK1 and ROCK2 play a role in pulmonary fibrosis. Studies on ROCK1+ / - and ROCK2+ / - mice with pulmonary fibrosis have shown that both subtypes exhibit protection against vascular leakage, myofibroblast differentiation, and fibrosis, and ROCK1+ / - mice show a greater attenuation of epithelial cell apoptosis. ROCK kinase is activated in the lungs of IPF patients and disease-related animal models, while ROCK kinase inhibitors can prevent tissue fibrosis in the model and induce the regression of established fibrosis. Regarding the safety of inhibiting ROCK1 and ROCK2, there is evidence that although ROCK inhibitors can dilate blood vessels, they do not necessarily induce systemic hypotension. Therefore, ROCK kinase inhibitors have great potential for the treatment of idiopathic pulmonary fibrosis.

[0005] Currently, 3 ROCK inhibitors have been marketed globally, including fasudil, ripasudil, and netarsudil. The indications for these 3 drugs are cerebral vasospasm, glaucoma, and high intraocular pressure. There is no ROCK inhibitor with an indication for IPF on the market yet.

[0006] The development of new drugs requires the careful optimization of the chemical and biological properties of lead compounds. Further, the compounds must have the desired pharmacokinetic and pharmacodynamic characteristics. This arduous development process usually requires extensive testing. In many cases, the process of determining the optimal compound often requires the preparation of thousands of structurally similar compounds. Therefore, improving ROCK kinase inhibitors and developing new backbone compounds with ROCK1 and / or ROCK2 kinase inhibitory effects are of positive significance for the treatment of the above diseases. At the same time, developing pharmaceutical forms suitable for these compounds to be made into drugs, such as forms that improve stability (e.g., storage stability), hygroscopicity, and / or efficacy (e.g., in vivo bioavailability), so as to achieve good results in the pharmaceutical and drug use stages, has become a technical problem to be solved urgently. Summary of the Invention

[0007] The present invention provides a salt of Compound I:

[0008]

[0009] The salt is an acid addition salt. For example, the salt is an acid addition salt of Compound I and any one of the following acids: hydrochloric acid, sulfuric acid, p-toluenesulfonic acid, benzenesulfonic acid, maleic acid, tartaric acid (including L-tartaric acid or R-tartaric acid), oxalic acid, ethanesulfonic acid, 2-hydroxyethanesulfonic acid, camphorsulfonic acid, 1,5-naphthalenedisulfonic acid; preferably sulfuric acid, tartaric acid (including L-tartaric acid or R-tartaric acid), ethanesulfonic acid or 2-hydroxyethanesulfonic acid.

[0010] According to an embodiment of the present invention, the hydrochloride salt of Compound I is an acid addition salt of Compound I and hydrochloric acid; preferably, the molar ratio of Compound I to hydrochloric acid is 1:(0.9 - 1.2), for example 1:1.

[0011] According to an embodiment of the present invention, the sulfate salt of Compound I is an acid addition salt of Compound I and sulfuric acid; preferably, the molar ratio of Compound I to sulfuric acid is 1:(0.4 - 0.6), for example 1:0.5.

[0012] According to an embodiment of the present invention, the p-toluenesulfonate salt of Compound I is an acid addition salt of Compound I and p-toluenesulfonic acid; preferably, the molar ratio of Compound I to p-toluenesulfonic acid is 1:(0.9 - 1.2), for example 1:1.

[0013] According to an embodiment of the present invention, the maleate salt of Compound I is an acid addition salt of Compound I and maleic acid; preferably, the molar ratio of Compound I to maleic acid is 1:(0.9 - 1.2), for example 1:1.

[0014] According to an embodiment of the present invention, the oxalate salt of Compound I is an acid addition salt of Compound I and oxalic acid; preferably, the molar ratio of Compound I to oxalic acid is 1:(0.9 - 1.2), for example 1:1.

[0015] According to an embodiment of the present invention, the camphorsulfonate salt of Compound I is an acid addition salt of Compound I and camphorsulfonic acid; preferably, the molar ratio of Compound I to camphorsulfonic acid is 1:(0.9 - 1.2), for example 1:1.

[0016] According to an embodiment of the present invention, the 2-hydroxyethanesulfonate salt of Compound I is an acid addition salt of Compound I and 2-hydroxyethanesulfonic acid; preferably, the molar ratio of Compound I to 2-hydroxyethanesulfonic acid is 1:(0.9 - 1.2), for example 1:1.

[0017] According to an embodiment of the present invention, the ethanesulfonate of Compound I is an acid addition salt of Compound I and ethanesulfonic acid; preferably, the molar ratio of Compound I to ethanesulfonic acid can be 1:(0.9 - 1.2), such as 1:1, 1:1.05.

[0018] According to an embodiment of the present invention, the tartrate of Compound I is an acid addition salt of Compound I and tartaric acid; preferably, the molar ratio of Compound I to tartaric acid can be 1:(0.9 - 1.2), such as 1:1. Among them, the tartaric acid is L-tartaric acid or R-tartaric acid.

[0019] According to an embodiment of the present invention, the 1,5-naphthalenedisulfonate of Compound I is an acid addition salt of Compound I and 1,5-naphthalenedisulfonic acid; preferably, the molar ratio of Compound I to 1,5-naphthalenedisulfonic acid can be 1:(0.8 - 1.1), such as 1:0.9.

[0020] According to an embodiment of the present invention, the benzenesulfonate of Compound I is an acid addition salt of Compound I and benzenesulfonic acid; preferably, the molar ratio of Compound I to benzenesulfonic acid can be 1:(0.9 - 1.2), such as 1:1.

[0021] According to an embodiment of the present invention, the salt of Compound I may contain crystal water or not (for example, it is an organic solvent compound). For example, it contains 1, 1.5, 2, 2.5, 3 or more crystal waters.

[0022] According to an embodiment of the present invention, the salt of Compound I can be amorphous or crystalline.

[0023] The present invention also provides the free base crystal form of the above Compound I.

[0024] According to an embodiment of the present invention, the free base crystal form is free base crystal form I.

[0025] According to an embodiment of the present invention, the free base crystal form I has characteristic peaks at 13.1 ± 0.2°, 20.5 ± 0.2° in the X-ray powder diffraction expressed in 2θ angle using Cu-Kα radiation; further, it also has a characteristic peak at 8.4 ± 0.2°; still further, it also has characteristic peaks at 6.2 ± 0.2°, 6.5 ± 0.2°, 10.9 ± 0.2°, 12.3 ± 0.2°, 14.1 ± 0.2°, 14.4 ± 0.2°, 24.2 ± 0.2°, 25.3 ± 0.2°.

[0026] According to an embodiment of the present invention, the free base crystal form I has substantially as Figure 1-1 the shown X-ray powder diffraction pattern.

[0027] According to an embodiment of the present invention, the free base crystalline form I has an X-ray powder diffraction expressed in 2θ angle using Cu-Kα radiation as Figure 1-2 shown, with an error range of ±0.2°.

[0028] According to an embodiment of the present invention, the free base crystalline form I is an anhydrate.

[0029] According to an embodiment of the present invention, the free base crystalline form I has almost no weight loss before the temperature of 150 ± 2 °C.

[0030] According to an embodiment of the present invention, the free base crystalline form I has only one endothermic peak, and the peak temperature is about 273 ± 5 °C, for example, 273 ± 2 °C.

[0031] Preferably, the free base crystalline form I has a DSC-TGA spectrum substantially as Figure 1-3 shown.

[0032] According to an embodiment of the present invention, the free base crystalline form I has residual organic solvents. For example, the organic solvents are dichloromethane and / or tetrahydrofuran. For example, the content of the organic solvents is 0.1-3%, for example, 1.2%. Exemplarily, it contains 0.5% of dichloromethane and 0.7% of tetrahydrofuran.

[0033] According to an embodiment of the present invention, the free base crystalline form I has a Figure 1-4 substantially as 1 1H-NMR spectrum shown.

[0034] The present invention also provides a crystalline form of the salt of the above compound I.

[0035] According to an embodiment of the present invention, the crystalline form of the salt is selected from the crystalline forms of the acid addition salts of compound I.

[0036] According to an embodiment of the present invention, the crystalline form of the salt is selected from one, two or more of the hydrochloride crystalline form, sulfate crystalline form, p-toluenesulfonate crystalline form, benzenesulfonate crystalline form, maleate crystalline form, oxalate crystalline form, camphorsulfonate crystalline form, 2-hydroxyethanesulfonate crystalline form, ethanesulfonate crystalline form, tartrate crystalline form, 1,5-naphthalenedisulfonate crystalline form of the above compound I.

[0037] According to an embodiment of the present invention, the crystalline form of the salt may contain or not contain solvent 1. For example, the solvent 1 is selected from organic solvent 1 or water.

[0038] According to an embodiment of the present invention, the organic solvent 1 is selected from one, two or more of methanol, ethanol, isopropanol, butanol, acetone, butanone, ethyl acetate, isopropyl acetate, methyl tert-butyl ether, dichloromethane, acetonitrile, tetrahydrofuran, n-heptane, dimethyl sulfoxide, 2-methyl-tetrahydrofuran, chloroform.

[0039] According to an embodiment of the present invention, the water may be water of crystallization or non-crystallized water.

[0040] According to an embodiment of the present invention, the hydrochloride crystal form is hydrochloride crystal form I.

[0041] According to an embodiment of the present invention, for the hydrochloride crystal form I, using Cu-Kα radiation, the X-ray powder diffraction expressed in terms of 2θ angle has characteristic peaks at 13.1±0.2°, 22.6±0.2°, 23.0±0.2°, and 24.4±0.2°; further, it also has characteristic peaks at 8.0±0.2°, 8.3±0.2°, 18.2±0.2°, 18.7±0.2°, and 30.3±0.2°; still further, it also has characteristic peaks at 15.8±0.2°, 16.2±0.2°, and 21.5±0.2°.

[0042] According to an embodiment of the present invention, the hydrochloride crystal form I has substantially as Figure 2-1 the X-ray powder diffraction pattern shown.

[0043] According to an embodiment of the present invention, for the hydrochloride crystal form I, using Cu-Kα radiation, the X-ray powder diffraction expressed in terms of 2θ angle is as Figure 2-2 shown, with an error range of ±0.2°.

[0044] According to an embodiment of the present invention, the hydrochloride crystal form I is an anhydrate.

[0045] According to an embodiment of the present invention, the hydrochloride crystal form I has a weight loss of about 0.1-1.0%, such as 0.3%, in the temperature range from room temperature to about 90±2°C.

[0046] According to an embodiment of the present invention, the hydrochloride crystal form I has two endothermic peaks, and the peak temperatures are approximately 56±5°C and 241±5°C respectively, for example, having endothermic peaks at 56±2°C and 241±2°C respectively.

[0047] Preferably, the hydrochloride crystal form I has substantially as Figure 2-3 the DSC-TGA pattern shown.

[0048] According to an embodiment of the present invention, the size of the hydrochloride crystal form I is <10 μm. For example, in one embodiment, the hydrochloride crystal form I has substantially as Figure 2-4 the polarized light microscopic morphology diagram shown.

[0049] According to an embodiment of the present invention, the hydrochloride crystal form I has no residual organic solvents.

[0050] According to an embodiment of the present invention, the hydrochloride crystal form I has substantially as Figure 2-5 shown 1 1H-NMR spectrum.

[0051] According to an embodiment of the present invention, the sulfate crystal form is sulfate crystal form I.

[0052] According to an embodiment of the present invention, for the sulfate crystal form I using Cu-Kα radiation, the X-ray powder diffraction expressed in 2θ angle has characteristic peaks at 8.4±0.2°, 11.0±0.2°, 13.7±0.2°, 19.3±0.2°; further, it also has characteristic peaks at 5.3±0.2°, 16.2±0.2°, 18.0±0.2°; still further, it also has characteristic peaks at 10.6±0.2°, 14.5±0.2°, 17.0±0.2°, 18.8±0.2°, 19.6±0.2°, 22.3±0.2°, 24.9±0.2°.

[0053] According to an embodiment of the present invention, the sulfate crystal form I has substantially as Figure 3-1 shown X-ray powder diffraction pattern.

[0054] According to an embodiment of the present invention, for the sulfate crystal form I using Cu-Kα radiation, the X-ray powder diffraction expressed in 2θ angle is as Figure 3-2 shown, with an error range of ±0.2°.

[0055] According to an embodiment of the present invention, the sulfate crystal form I is a hydrate, such as a monohydrate, dihydrate, trihydrate, preferably a monohydrate.

[0056] According to an embodiment of the present invention, the sulfate crystal form I loses weight by about 0.1 - 5.0% in the temperature range from room temperature to about 125±2°C, for example 3%.

[0057] According to an embodiment of the present invention, the sulfate crystal form I has two endothermic peaks, and the peak temperatures are approximately 104±5°C and 163±5°C respectively, for example 104±2°C and 163±2°C respectively.

[0058] Preferably, the sulfate crystal form I has substantially as Figure 3-3 the DSC-TGA pattern shown.

[0059] According to an embodiment of the present invention, the sulfate crystal form I is an irregular crystal. For example, in one embodiment, the sulfate crystal form I has substantially as Figure 3-4 shown polarized light microscopic morphology diagram.

[0060] According to an embodiment of the present invention, the sulfate crystal form I has no residual organic solvents.

[0061] According to an embodiment of the present invention, the sulfate crystal form I has a Figure 3-5 shown 1 1H-NMR spectrum.

[0062] According to an embodiment of the present invention, the p-toluenesulfonate crystal form is p-toluenesulfonate crystal form I.

[0063] According to an embodiment of the present invention, for the p-toluenesulfonate crystal form I using Cu-Kα radiation, the X-ray powder diffraction expressed in 2θ angles has characteristic peaks at 5.1±0.2°, 26.1±0.2°; further, it also has characteristic peaks at 11.8±0.2°, 26.5±0.2°; still further, it also has characteristic peaks at 10.2±0.2°, 11.0±0.2°, 18.9±0.2°, 19.2±0.2°, 21.0±0.2°.

[0064] According to an embodiment of the present invention, the p-toluenesulfonate crystal form I has a Figure 4-1 shown X-ray powder diffraction pattern.

[0065] According to an embodiment of the present invention, for the p-toluenesulfonate crystal form I using Cu-Kα radiation, the X-ray powder diffraction expressed in 2θ angles is Figure 4-2 shown, with an error range of ±0.2°.

[0066] According to an embodiment of the present invention, the p-toluenesulfonate crystal form I is a hydrate, such as 0.5 hydrate, 0.75 hydrate or monohydrate, preferably 0.75 hydrate or monohydrate.

[0067] According to an embodiment of the present invention, the p-toluenesulfonate crystal form I loses about 0.5 - 5%, such as 2%, of its weight in the temperature range from room temperature to about 110±2°C.

[0068] According to an embodiment of the present invention, the p-toluenesulfonate crystal form I has two endothermic peaks, and the peak temperatures are approximately 94±5°C and 198±5°C respectively, such as 94±2°C and 198±2°C respectively.

[0069] Preferably, the p-toluenesulfonate crystal form I has a Figure 4-3 shown DSC-TGA spectrum.

[0070] According to an embodiment of the present invention, the p-toluenesulfonate crystal form I is an irregular crystal. For example, in one embodiment, the p-toluenesulfonate crystal form I has a Figure 4-4 shown polarized light microscopic morphology diagram.

[0071] According to an embodiment of the present invention, the p-toluenesulfonate polymorph I has substantially as Figure 4-5 shown 1 1H-NMR spectrum, with (trace amounts of) residual organic solvents.

[0072] According to an embodiment of the present invention, the benzenesulfonate polymorph is benzenesulfonate polymorph I.

[0073] According to an embodiment of the present invention, for the benzenesulfonate polymorph I, using Cu-Kα radiation, the X-ray powder diffraction expressed in 2θ angles has characteristic peaks at 6.0±0.2°, 12.0±0.2°; further, it also has characteristic peaks at 13.2±0.2°, 18.2±0.2°, 22.2±0.2°; still further, it also has characteristic peaks at 8.7±0.2°, 17.4±0.2°, 18.6±0.2°, 19.4±0.2°, 20.3±0.2°, 24.2±0.2°, 30.5±0.2°.

[0074] According to an embodiment of the present invention, the benzenesulfonate polymorph I has substantially as Figure 5-1 shown X-ray powder diffraction pattern.

[0075] According to an embodiment of the present invention, for the benzenesulfonate polymorph I, using Cu-Kα radiation, the X-ray powder diffraction expressed in 2θ angles is as Figure 5-2 shown, with an error range of ±0.2°.

[0076] According to an embodiment of the present invention, the benzenesulfonate polymorph I is a hydrate, such as a monohydrate, 1.5-hydrate, dihydrate or trihydrate, preferably 1.5-hydrate or monohydrate.

[0077] According to an embodiment of the present invention, the benzenesulfonate polymorph I loses weight by about 1-5%, such as 3.9%, in the temperature range from room temperature to about 100±2°C.

[0078] According to an embodiment of the present invention, the benzenesulfonate polymorph I has two endothermic peaks, and the peak temperatures are approximately 109±5°C and 176±5°C respectively, such as 109±2°C and 176±2°C respectively.

[0079] Preferably, the benzenesulfonate polymorph I has substantially as Figure 5-3 the DSC-TGA pattern as described.

[0080] According to an embodiment of the present invention, the benzenesulfonate polymorph I is a microcrystal, such as with a size less than 20μm, and also such as with a size less than 10μm. For example, in one embodiment, the benzenesulfonate polymorph I has substantially as Figure 5-4 shown polarized light microscopic morphology diagram.

[0081] According to an embodiment of the present invention, the benzenesulfonate polymorph I has substantially as Figure 5-5 shown 1 an H-NMR spectrum.

[0082] According to an embodiment of the present invention, the maleate polymorphs include maleate polymorph I, maleate polymorph II, and maleate polymorph III.

[0083] According to an embodiment of the present invention, the maleate polymorph I, using Cu-Kα radiation, has characteristic peaks at 24.8 ± 0.2° in terms of 2θ angle in X-ray powder diffraction; further, it also has characteristic peaks at 12.0 ± 0.2° and 20.5 ± 0.2°; still further, it also has characteristic peaks at 9.3 ± 0.2°, 13.2 ± 0.2°, 17.4 ± 0.2°, and 20.0 ± 0.2°.

[0084] According to an embodiment of the present invention, the maleate polymorph I has substantially as Figure 6-1 shown X-ray powder diffraction pattern.

[0085] According to an embodiment of the present invention, the maleate polymorph I, using Cu-Kα radiation, has X-ray powder diffraction as Figure 6-2 shown, with an error range of ±0.2°.

[0086] According to an embodiment of the present invention, the maleate polymorph I is a solvate, preferably an organic solvent solvate, such as a methanol solvate, an ethanol solvate, or an isopropanol solvate. For example, the amount of the organic solvent is 2.0 - 4.0%, such as 2.9%.

[0087] According to an embodiment of the present invention, the maleate polymorph I loses weight by about 0.5 - 5%, such as 2.6%, in the temperature range from room temperature to about 180 ± 2°C.

[0088] According to an embodiment of the present invention, the maleate polymorph I has an endothermic peak at a peak temperature of about 198 ± 5°C.

[0089] Preferably, the maleate polymorph I has substantially as Figure 6-3 the described DSC-TGA pattern.

[0090] According to an embodiment of the present invention, the maleate polymorph I is a microcrystal. For example, in one embodiment, the size of the maleate polymorph I is less than 20 μm, such as less than 10 μm. For example, in one embodiment, the maleate polymorph I has substantially as Figure 6-4 shown polarized light microscopic morphology.

[0091] According to an embodiment of the present invention, the maleate polymorph I has substantially as Figure 6-5 shown 1 1H-NMR spectrum.

[0092] According to an embodiment of the present invention, the maleate polymorph II has characteristic peaks at 11.0±0.2°, 14.4±0.2°, 16.2±0.2°, 16.8±0.2° in the X-ray powder diffraction expressed in 2θ angle using Cu-Kα radiation; further, it also has characteristic peaks at 5.4±0.2°, 12.1±0.2°, 13.4±0.2°, 17.2±0.2°, 20.3±0.2°, 20.6±0.2°; still further, it also has characteristic peaks at 7.2±0.2°, 17.8±0.2°, 25.0±0.2°.

[0093] According to an embodiment of the present invention, the maleate polymorph II has substantially as Figure 7-1 shown X-ray powder diffraction pattern.

[0094] According to an embodiment of the present invention, the maleate polymorph II has X-ray powder diffraction expressed in 2θ angle using Cu-Kα radiation as Figure 7-2 shown, with an error range of ±0.2°.

[0095] According to an embodiment of the present invention, the maleate polymorph II is an anhydrate.

[0096] According to an embodiment of the present invention, the maleate polymorph II has no obvious weight loss in the temperature range from room temperature to about 100±2°C.

[0097] According to an embodiment of the present invention, the maleate polymorph II has two endothermic peaks, and the peak temperatures are approximately 48±5°C and 190±5°C respectively, for example 48±2°C and 190±2°C respectively.

[0098] Preferably, the maleate polymorph II has substantially as Figure 7-3 the DSC-TGA spectrum as described.

[0099] According to an embodiment of the present invention, the maleate polymorph II is in the form of agglomerated particles. Preferably, the maleate polymorph II has substantially as Figure 7-4 shown polarized light microscopic morphology diagram.

[0100] According to an embodiment of the present invention, the maleate polymorph II has organic solvent residues. For example, the residue amount of the organic solvent is 2.0 - 3.5%, for example 2.8%. For example, the organic solvent is one, two or more (such as three) of acetonitrile, tetrahydrofuran, isopropanol, isopropyl acetate, methyl ethyl ketone.

[0101] According to an embodiment of the present invention, the maleate polymorph II has a Figure 7-5 as shown 1 1H-NMR spectrum.

[0102] According to an embodiment of the present invention, for the maleate polymorph III, using Cu-Kα radiation, the X-ray powder diffraction expressed in 2θ angle has characteristic peaks at 14.6 ± 0.2°, 16.9 ± 0.2°, 25.8 ± 0.2°; further, it also has characteristic peaks at 5.3 ± 0.2°, 10.7 ± 0.2°, 26.3 ± 0.2°; still further, it also has characteristic peaks at 12.4 ± 0.2°, 16.1 ± 0.2°, 19.2 ± 0.2°, 20.2 ± 0.2°.

[0103] According to an embodiment of the present invention, the maleate polymorph III has a Figure 8-1 X-ray powder diffraction pattern as shown.

[0104] According to an embodiment of the present invention, for the maleate polymorph III, using Cu-Kα radiation, the X-ray powder diffraction expressed in 2θ angle is as Figure 8-2 shown, with an error range of ±0.2°.

[0105] According to an embodiment of the present invention, the maleate polymorph III is an anhydrate.

[0106] According to an embodiment of the present invention, the maleate polymorph III has no obvious weight loss in the temperature range from room temperature to about 150 ± 2 °C.

[0107] According to an embodiment of the present invention, the maleate polymorph III has an endothermic peak at a peak temperature of about 197 ± 5 °C.

[0108] Preferably, the maleate polymorph III has a Figure 8-3 DSC-TGA pattern as described.

[0109] According to an embodiment of the present invention, the maleate polymorph III is composed of microcrystals. For example, in one embodiment, the maleate polymorph III has a Figure 8-4 polarized light microscopic morphology diagram as shown.

[0110] According to an embodiment of the present invention, the maleate polymorph III has residual organic solvents. For example, the residual amount of the organic solvent is 0.05 - 1%, such as 0.5%. For example, the organic solvent is one, two or more (such as three) of acetone, isopropyl acetate, butanol, acetonitrile, and butanone.

[0111] According to an embodiment of the present invention, the maleate polymorph III has substantially as Figure 8-5 shown 1 1H-NMR spectrum.

[0112] According to an embodiment of the present invention, the oxalate polymorph is oxalate polymorph I.

[0113] According to an embodiment of the present invention, the oxalate polymorph I has characteristic peaks at 18.4±0.2° and 24.3±0.2° in the X-ray powder diffraction expressed in 2θ angle using Cu-Kα radiation; further, it also has characteristic peaks at 10.6±0.2° and 15.2±0.2°; still further, it also has characteristic peaks at 7.6±0.2°, 11.2±0.2°, and 12.6±0.2°.

[0114] According to an embodiment of the present invention, the oxalate polymorph I has substantially as Figure 9-1 shown X-ray powder diffraction pattern.

[0115] According to an embodiment of the present invention, the oxalate polymorph I has X-ray powder diffraction expressed in 2θ angle using Cu-Kα radiation as Figure 9-2 shown, with an error range of ±0.2°.

[0116] According to an embodiment of the present invention, the oxalate polymorph I is a solvate, such as an organic solvate, preferably a methanol solvate, an ethanol solvate, or an isopropanol solvate. For example, the amount of the organic solvent is 2.0 to 5.0%, such as 3.9%.

[0117] According to an embodiment of the present invention, the oxalate polymorph I loses about 2 to 6% of its weight in the temperature range of 60-160°C, such as 4.1%.

[0118] According to an embodiment of the present invention, the oxalate polymorph I has a broad endothermic peak at a peak temperature of about 127±5°C.

[0119] Preferably, the oxalate polymorph I has substantially as Figure 9-3 the DSC-TGA spectrum as described.

[0120] According to an embodiment of the present invention, the oxalate polymorph I is a microcrystal. For example, in one embodiment, the oxalate polymorph I has substantially as Figure 9-4 shown polarized light microscopic morphology.

[0121] According to an embodiment of the present invention, the oxalate polymorph I has substantially as Figure 9-5 shown 1 1H-NMR spectrum.

[0122] According to an embodiment of the present invention, the camphorsulfonate crystal form is camphorsulfonate crystal form I.

[0123] According to an embodiment of the present invention, for the camphorsulfonate crystal form I, using Cu-Kα radiation, the X-ray powder diffraction expressed in 2θ angle has characteristic peaks at 3.6±0.2°, 12.6±0.2°, and 16.5±0.2°; further, it also has characteristic peaks at 9.6±0.2°, 14.8±0.2°, 17.4±0.2°, and 19.0±0.2°; still further, it also has characteristic peaks at 7.3±0.2°, 20.2±0.2°, 21.5±0.2°, 21.8±0.2°, 23.8±0.2°, 25.4±0.2°, and 28.1±0.2°.

[0124] According to an embodiment of the present invention, the camphorsulfonate crystal form I has substantially as Figure 10-1 the shown X-ray powder diffraction pattern.

[0125] According to an embodiment of the present invention, the camphorsulfonate crystal form I has the X-ray powder diffraction characteristic peaks expressed in 2θ angle as shown in Table 1, with an error range of ±0.2°:

[0126] Table 1

[0127]

[0128] . Preferably, the camphorsulfonate crystal form I has the X-ray powder diffraction characteristic peaks expressed in 2θ angle as shown in Table 1', with an error range of ±0.2°.

[0129] According to an embodiment of the present invention, the camphorsulfonate crystal form I is a hydrate, such as a monohydrate, a dihydrate, or a trihydrate, preferably a dihydrate.

[0130] According to an embodiment of the present invention, the camphorsulfonate crystal form I loses about 5% of its weight in the temperature range from room temperature to 180±2°C.

[0131] According to an embodiment of the present invention, the camphorsulfonate crystal form I has two endothermic peaks, and the peak temperatures are approximately 83±5°C and 198±5°C respectively, for example, 83±2°C and 198±2°C respectively.

[0132] Preferably, the camphorsulfonate crystal form I has substantially as Figure 10-2 the described DSC-TGA pattern.

[0133] According to an embodiment of the present invention, the camphorsulfonate crystal form I has no residual organic solvents.

[0134] According to an embodiment of the present invention, the camphorsulfonate polymorph I has substantially as Figure 10-3 shown 1 1H-NMR spectrum.

[0135] According to an embodiment of the present invention, the 2-hydroxyethanesulfonate polymorph is 2-hydroxyethanesulfonate polymorph I.

[0136] According to an embodiment of the present invention, the 2-hydroxyethanesulfonate polymorph I has characteristic peaks at 16.0 ± 0.2°, 25.4 ± 0.2° in terms of 2θ angle in X-ray powder diffraction using Cu-Kα radiation; further, it also has characteristic peaks at 18.0 ± 0.2°, 18.8 ± 0.2°; still further, it also has characteristic peaks at 8.5 ± 0.2°, 8.9 ± 0.2°, 13.2 ± 0.2°; even further, it also has characteristic peaks at 15.4 ± 0.2°, 20.7 ± 0.2°, 22.3 ± 0.2°, 22.6 ± 0.2°, 24.7 ± 0.2°, 26.1 ± 0.2°.

[0137] According to an embodiment of the present invention, the 2-hydroxyethanesulfonate polymorph I has substantially as Figure 11-1 shown X-ray powder diffraction pattern.

[0138] According to an embodiment of the present invention, the 2-hydroxyethanesulfonate polymorph I has X-ray powder diffraction characteristic peaks shown in Table 2 in terms of 2θ angle, with an error range of ±0.2°:

[0139] Table 2

[0140]

[0141] . Preferably, the 2-hydroxyethanesulfonate polymorph I has X-ray powder diffraction characteristic peaks shown in Table 2' in terms of 2θ angle, with an error range of ±0.2°.

[0142] According to an embodiment of the present invention, the 2-hydroxyethanesulfonate polymorph I is an anhydrate.

[0143] According to an embodiment of the present invention, the 2-hydroxyethanesulfonate polymorph I has a weight loss of 0 to 0.5% in the temperature range from room temperature to 150 ± 2°C, for example, almost no weight loss or a weight loss of 0.18 ± 0.02%; and a weight loss of 5.4 ± 0.2% in the temperature range from room temperature to 300 ± 2°C, for example, 5.4%.

[0144] According to an embodiment of the present invention, the 2-hydroxyethanesulfonate polymorph I has an exothermic peak at a peak temperature of about 258 ± 5°C.

[0145] Preferably, the 2-hydroxyethanesulfonate polymorph I has substantially asFigure 11-2 The DSC-TGA spectrum described above.

[0146] According to an embodiment of the present invention, the 2-hydroxyethanesulfonate polymorph I has no residual organic solvent.

[0147] According to an embodiment of the present invention, the 2-hydroxyethanesulfonate polymorph I has substantially as Figure 11-3 shown 1 1H-NMR spectrum.

[0148] According to an embodiment of the present invention, the ethanesulfonate polymorphs include ethanesulfonate polymorph I and ethanesulfonate polymorph II.

[0149] According to an embodiment of the present invention, for the ethanesulfonate polymorph I using Cu-Kα radiation, the X-ray powder diffraction expressed in 2θ angle has characteristic peaks at 24.6±0.2°; further, it also has characteristic peaks at 10.4±0.2° and 14.8±0.2°; still further, it also has characteristic peaks at 16.9±0.2°, 19.3±0.2°, and 21.4±0.2°; even further, it also has characteristic peaks at 15.8±0.2°, 17.4±0.2°, 20.5±0.2°, 23.0±0.2°, and 26.2±0.2°.

[0150] According to an embodiment of the present invention, the ethanesulfonate polymorph I has substantially as Figure 12-1-1 or Figure 12-1-2 shown X-ray powder diffraction pattern.

[0151] According to an embodiment of the present invention, the ethanesulfonate polymorph I has the X-ray powder diffraction characteristic peaks expressed in 2θ angle as shown in Table 4, with an error range of ±0.2°:

[0152] Table 4

[0153]

[0154] . Preferably, the ethanesulfonate polymorph I has the X-ray powder diffraction characteristic peaks expressed in 2θ angle as shown in Table 4', with an error range of ±0.2°.

[0155] According to still another embodiment of the present invention, the ethanesulfonate polymorph I has the X-ray powder diffraction characteristic peaks expressed in 2θ angle as shown in Table 5, with an error range of ±0.2°:

[0156] Table 5

[0157]

[0158] Preferably, the etilsulfonate polymorph I has X-ray powder diffraction characteristic peaks expressed in 2θ angles as shown in Table 6’, with an error range of ±0.2°.

[0159] According to an embodiment of the present invention, the etilsulfonate polymorph I is an anhydrate.

[0160] According to an embodiment of the present invention, the etilsulfonate polymorph I has a weight loss of about 1.4 ± 0.4% before 230 ± 2 °C, preferably a weight loss of 1.4 ± 0.2% before 230 ± 2 °C.

[0161] According to an embodiment of the present invention, the etilsulfonate polymorph I has an exothermic peak at a peak temperature of about 265 ± 5 °C, such as 265 ± 2 °C, 268 ± 2 °C.

[0162] According to an embodiment of the present invention, the etilsulfonate polymorph I has an endothermic peak at a peak temperature of about 260 ± 5 °C, such as 259 ± 2 °C, 263 ± 2 °C.

[0163] The peak temperatures of the exothermic peak and the endothermic peak of the above etilsulfonate polymorph I are different, and preferably the peak temperature of the endothermic peak < the peak temperature of the exothermic peak.

[0164] Preferably, the etilsulfonate polymorph I has substantially as Figure 12-2-1 or the DSC-TGA spectrum shown in 12-2-2.

[0165] According to an embodiment of the present invention, the etilsulfonate polymorph I has no residual organic solvent, or the etilsulfonate polymorph I has residual organic solvent. For example, the content of the organic solvent is 0.1-2.5%, such as 0.9-1.6%, and exemplarily 1.3%. For example, the organic solvent is selected from one, two or more of methanol, ethanol, isopropanol, acetonitrile, tetrahydrofuran, methyl ethyl ketone, acetone, 2-methyltetrahydrofuran, isopropyl acetate, ethyl acetate, methyl tert-butyl ether, dimethyl sulfoxide, and preferably methanol, isopropanol, ethanol, acetonitrile or methyl ethyl ketone. Exemplarily, it contains about 1.3% of ethanol.

[0166] According to an embodiment of the present invention, the etilsulfonate polymorph I is a microcrystal. For example, in one embodiment, the etilsulfonate polymorph I has substantially as Figure 12-3-5 the polarized light microscopic morphology diagram shown.

[0167] According to an embodiment of the present invention, the etilsulfonate polymorph I has substantially as Figure 12-3-1 or the 1 1H-NMR diagram shown in 12-3-2 or 12-3-3.

[0168] According to an embodiment of the present invention, the etilsulfonate polymorph II has characteristic peaks in an X-ray powder diffraction represented by 2θ angles at 10.6±0.2°, 17.5±0.2° using Cu-Kα radiation; further, it also has characteristic peaks at 4.5±0.2°, 9.0±0.2°, 19.2±0.2°; still further, it also has characteristic peaks at 13.5±0.2°, 14.7±0.2°, 16.2±0.2°, 18.0±0.2°, 22.7±0.2°, 25.2±0.2°, 26.9±0.2°.

[0169] According to an embodiment of the present invention, the etilsulfonate polymorph II has an X-ray powder diffraction pattern substantially as Figure 13-1 shown.

[0170] According to an embodiment of the present invention, the etilsulfonate polymorph II has X-ray powder diffraction characteristic peaks represented by 2θ angles as shown in Table 6, with an error range of ±0.2°:

[0171] Table 6

[0172]

[0173] . Preferably, the etilsulfonate polymorph II has X-ray powder diffraction characteristic peaks represented by 2θ angles as shown in Table 5', with an error range of ±0.2°.

[0174] According to an embodiment of the present invention, the etilsulfonate polymorph II is a solvate, such as an organic solvent solvate or a hydrate. For example, the organic solvent is selected from one, two or more of ethanol, acetonitrile, tetrahydrofuran, methyl ethyl ketone, acetone, 2-methyltetrahydrofuran, isopropyl acetate, ethyl acetate, methyl tert-butyl ether. For example, the content of the organic solvent is 1.5 to 3.5%, such as 2.3%. Exemplarily, it contains about 2.3% isopropyl acetate.

[0175] For example, the hydrate is a monohydrate, a 0.5 hydrate, a dihydrate, preferably a monohydrate.

[0176] According to an embodiment of the present invention, the etilsulfonate polymorph II has a weight loss of about 2.3±0.2% before 150±2°C.

[0177] According to an embodiment of the present invention, the etilsulfonate polymorph II has an endothermic peak at a peak temperature of about 85±5°C, an endothermic peak at a peak temperature of about 177±5°C, and an exothermic peak at a peak temperature of about 222±5°C. For example, the etilsulfonate polymorph II has an endothermic peak at a peak temperature of about 85±2°C, an endothermic peak at a peak temperature of about 177±2°C, and an exothermic peak at a peak temperature of about 222±2°C.

[0178] Preferably, the etanesulfonate polymorph II has substantially as Figure 13-2 the DSC-TGA spectrum as described.

[0179] According to an embodiment of the present invention, the tartrate polymorph includes L-tartrate polymorph, D-tartrate polymorph, preferably L-tartrate polymorph I.

[0180] According to an embodiment of the present invention, for the L-tartrate polymorph I, using Cu-Kα radiation, the X-ray powder diffraction expressed in 2θ angle has characteristic peaks at 11.6±0.2°, 15.5±0.2°, 20.8±0.2°; further, it also has characteristic peaks at 9.2±0.2°, 18.7±0.2°, 21.1±0.2°; still further, it also has characteristic peaks at 7.2±0.2°, 7.7±0.2°, 12.1±0.2°, 14.5±0.2°, 19.4±0.2°, 22.6±0.2°.

[0181] According to an embodiment of the present invention, the L-tartrate polymorph I has substantially as Figure 14-1 the X-ray powder diffraction spectrum as shown.

[0182] According to an embodiment of the present invention, the L-tartrate polymorph I has the X-ray powder diffraction characteristic peaks expressed in 2θ angle as shown in Table 11-3, with an error range of ±0.2°.

[0183] According to an embodiment of the present invention, the L-tartrate polymorph I is an anhydrate.

[0184] According to an embodiment of the present invention, the L-tartrate polymorph I has a weight loss of about 2-10% in the temperature range from room temperature to 150±2°C, such as a weight loss of 3.6±0.2% and 6.8±0.2%.

[0185] According to an embodiment of the present invention, the L-tartrate polymorph I has an endothermic peak at a peak temperature of about 213±5°C.

[0186] Preferably, the L-tartrate polymorph I has substantially as Figure 14-2 the DSC-TGA spectrum as shown.

[0187] According to an embodiment of the present invention, the L-tartrate polymorph I has residual organic solvents. For example, the content of the organic solvent is 0.3-1.5%, for example, it is 0.7%. Exemplarily, it contains 0.7% of methyl ethyl ketone or ethyl acetate.

[0188] According to an embodiment of the present invention, the L-tartrate polymorph I has substantially asFigure 14-3 as shown 1 H-NMR spectrum

[0189] According to an embodiment of the present invention, the 1,5-naphthalenedisulfonate crystal form includes 1,5-naphthalenedisulfonate crystal form I and 1,5-naphthalenedisulfonate crystal form II.

[0190] According to an embodiment of the present invention, for the 1,5-naphthalenedisulfonate crystal form I, using Cu-Kα radiation, the X-ray powder diffraction expressed in 2θ angle has characteristic peaks at 7.7±0.2°, 18.0±0.2°, 23.5±0.2°; further, it also has characteristic peaks at 15.4±0.2°, 24.0±0.2°, 24.8±0.2°; still further, the crystal form also has characteristic peaks at 8.0±0.2°, 8.6±0.2°, 10.8±0.2°, 11.8±0.2°, 16.8±0.2°, 17.7±0.2°, 19.8±0.2°, 22.5±0.2°; or the crystal form also has characteristic peaks at 8.0±0.2°, 8.6±0.2°, 10.9±0.2°, 11.9±0.2°, 16.8±0.2°, 17.7±0.2°, 19.8±0.2°, 22.6±0.2°.

[0191] According to an embodiment of the present invention, the 1,5-naphthalenedisulfonate crystal form I has an X-ray powder diffraction pattern substantially as Figure 15-1 shown

[0192] According to an embodiment of the present invention, the 1,5-naphthalenedisulfonate crystal form I has X-ray powder diffraction characteristic peaks expressed in 2θ angle as shown in Table 12-3, with an error range of ±0.2°.

[0193] According to an embodiment of the present invention, the 1,5-naphthalenedisulfonate crystal form I is a hydrate, such as monohydrate, dihydrate, trihydrate or tetrahydrate, preferably tetrahydrate.

[0194] According to an embodiment of the present invention, the 1,5-naphthalenedisulfonate crystal form I has a weight loss of about 2-10% in the temperature range from room temperature to 150±2°C, such as a weight loss of 8.3±0.2%.

[0195] According to an embodiment of the present invention, the 1,5-naphthalenedisulfonate crystal form I has two endothermic peaks, and the peak temperatures are approximately 84±5°C and 119±5°C respectively; for example, the peak temperatures are approximately 84.0±2°C and 119.6±2°C respectively.

[0196] Preferably, the 1,5-naphthalenedisulfonate crystal form I has a DSC-TGA pattern substantially as Figure 15-2 described

[0197] According to an embodiment of the present invention, the 1,5-naphthalenedisulfonate polymorph I has no residual organic solvents.

[0198] According to an embodiment of the present invention, the 1,5-naphthalenedisulfonate polymorph I has substantially as Figure 15-3 shown 1 1H-NMR spectrum.

[0199] According to an embodiment of the present invention, for the 1,5-naphthalenedisulfonate polymorph II, using Cu-Kα radiation, the X-ray powder diffraction expressed in 2θ angle has characteristic peaks at 25.5±0.2°, 10.2±0.2°; further, it also has characteristic peaks at 3.1±0.2°, 3.9±0.2°, 8.6±0.2°, 12.7±0.2°, 13.9±0.2°; still further, it also has characteristic peaks at 8.4±0.2°, 11.4±0.2°, 14.4±0.2°, 15.6±0.2°, 20.5±0.2°, 20.9±0.2°, 22.0±0.2°, 23.8±0.2°.

[0200] According to an embodiment of the present invention, the 1,5-naphthalenedisulfonate polymorph II has substantially as Figure 16-1 shown X-ray powder diffraction pattern.

[0201] According to an embodiment of the present invention, the 1,5-naphthalenedisulfonate polymorph II has the X-ray powder diffraction characteristic peaks shown in Table 12-4, with an error range of ±0.2°.

[0202] According to an embodiment of the present invention, the 1,5-naphthalenedisulfonate polymorph II is a hydrate, such as monohydrate, dihydrate, trihydrate or tetrahydrate, preferably trihydrate.

[0203] According to an embodiment of the present invention, the 1,5-naphthalenedisulfonate polymorph II has a weight loss of about 2-12% in the temperature range from room temperature to 180±2°C, such as a weight loss of 8.0±0.2%.

[0204] According to an embodiment of the present invention, the 1,5-naphthalenedisulfonate polymorph II has three endothermic peaks, and the peak temperatures are approximately 125±5°C, 170±5°C, 217±5°C respectively; for example, the peak temperatures are approximately 125.4±2°C, 170.5±2°C, 217.3±2°C respectively.

[0205] Preferably, the 1,5-naphthalenedisulfonate polymorph II has substantially as Figure 16-2 the shown DSC-TGA pattern.

[0206] According to an embodiment of the present invention, the 1,5-naphthalenedisulfonate polymorph II has residual organic solvents. For example, the mass content of the organic solvent is 0.2 to 1.0%, such as 0.6%. Exemplarily, the 1,5-naphthalenedisulfonate polymorph II contains methyl ethyl ketone.

[0207] According to an embodiment of the present invention, the 1,5-naphthalenedisulfonate polymorph II has substantially as Figure 16-3 shown 1 1H-NMR spectrum.

[0208] The present invention also provides a method for preparing a salt or a crystal form of a salt of the above compound I, comprising mixing and reacting compound I with an acid to obtain the salt or the crystal form of the salt of compound I.

[0209] According to an embodiment of the present invention, the acid has the selection as shown above.

[0210] According to an embodiment of the present invention, compound I and the acid have the molar ratio as described above.

[0211] According to an embodiment of the present invention, the reaction is carried out in solvent 2.

[0212] For example, solvent 2 is selected from one, two or more of organic solvent 2, water, and a mixed solvent of organic solvent 2 and water.

[0213] According to an embodiment of the present invention, organic solvent 2 is selected from one, two or more of methanol, ethanol, isopropanol, butanol (including n-butanol or isobutanol), acetone, methyl ethyl ketone, ethyl acetate, isopropyl acetate, methyl tert-butyl ether, dichloromethane, acetonitrile, tetrahydrofuran, n-heptane, dimethyl sulfoxide, 2-methyltetrahydrofuran, chloroform.

[0214] Preferably, organic solvent 2 is selected from one, two or more of methanol, ethanol, acetonitrile, tetrahydrofuran, isopropanol, acetone, butanol, methyl ethyl ketone, isopropyl acetate, ethyl acetate, dimethyl sulfoxide, 2-methyltetrahydrofuran.

[0215] According to an embodiment of the present invention, the solvents for preparing the hydrochloride or the crystal form of the hydrochloride of compound I (such as hydrochloride crystal form I) are selected from one, two or more of methanol, ethanol, isopropanol, acetonitrile, methyl ethyl ketone, tetrahydrofuran, 2-methyltetrahydrofuran (such as three), and preferably one, two or three of methanol, ethanol, isopropanol.

[0216] According to an embodiment of the present invention, the solvents for preparing the sulfate or the crystal form of the sulfate of compound I (such as sulfate crystal form I) are selected from one, two or more of methanol, ethanol, isopropanol, acetonitrile, methyl ethyl ketone, and preferably one, two or three of methanol, ethanol, isopropanol.

[0217] According to an embodiment of the present invention, the solvent for preparing the p-toluenesulfonate or p-toluenesulfonate crystal form (such as p-toluenesulfonate crystal form I) of Compound I is selected from one, two or more (such as three) of methanol, ethanol, isopropanol, acetonitrile, methyl ethyl ketone, tetrahydrofuran, 2-methyltetrahydrofuran, and is preferably one, two or three of methanol, ethanol, and isopropanol.

[0218] According to an embodiment of the present invention, the solvent for preparing the benzenesulfonate or benzenesulfonate crystal form (such as benzenesulfonate crystal form I) of Compound I is selected from one, two or more (such as three) of methanol, ethanol, isopropanol, acetonitrile, tetrahydrofuran, 2-methyltetrahydrofuran, and is preferably one, two or three of methanol, ethanol, and isopropanol.

[0219] According to an embodiment of the present invention, the solvent for preparing the maleate or maleate crystal form of Compound I is selected from one, two or more of methanol, ethanol, acetonitrile, tetrahydrofuran, isopropanol, acetone, butanol, methyl ethyl ketone, and isopropyl acetate.

[0220] Exemplarily, the solvent for preparing maleate crystal form I is one, two or three of methanol, ethanol, and isopropanol;

[0221] Exemplarily, the solvent for preparing maleate crystal form II is one, two or three of acetonitrile, tetrahydrofuran, isopropanol, isopropyl acetate, and methyl ethyl ketone;

[0222] Exemplarily, the solvent for preparing maleate crystal form III is one, two or three of acetone, isopropyl acetate, butanol, acetonitrile, and methyl ethyl ketone.

[0223] According to an embodiment of the present invention, the solvent for preparing the oxalate or oxalate crystal form (such as oxalate crystal form I) of Compound I is one, two or three of methanol, ethanol, and isopropanol.

[0224] According to an embodiment of the present invention, the solvent for preparing the camphorsulfonate or camphorsulfonate crystal form (such as camphorsulfonate crystal form I) of Compound I is selected from one, two or more of methanol, isopropanol, ethanol, acetonitrile, tetrahydrofuran, methyl ethyl ketone, isopropyl acetate, and ethyl acetate.

[0225] According to an embodiment of the present invention, the solvent for preparing the 2-hydroxyethanesulfonate or 2-hydroxyethanesulfonate crystal form (such as 2-hydroxyethanesulfonate crystal form I) of Compound I is selected from one, two or more of ethanol, acetonitrile, tetrahydrofuran, methyl ethyl ketone, and isopropyl acetate, and is preferably one, two or three of ethanol, methanol, and isopropanol.

[0226] According to an embodiment of the present invention, the solvent for preparing the ethanesulfonate or ethanesulfonate crystal form of Compound I is selected from one, two or more of ethanol, acetonitrile, tetrahydrofuran, methyl ethyl ketone, and isopropyl acetate.

[0227] Exemplarily, the solvent for preparing ethanesulfonate crystal form I is selected from one, two or more of dimethyl sulfoxide, methanol, isopropanol, ethyl acetate, isopropyl acetate, ethanol, acetonitrile, tetrahydrofuran, methyl ethyl ketone, acetone, and dichloromethane; preferably dimethyl sulfoxide, methanol, ethanol, isopropanol, acetonitrile, methyl ethyl ketone, or a mixed solvent of these solvents.

[0228] Exemplarily, the solvent for preparing ethanesulfonate crystal form II is selected from isopropyl acetate.

[0229] According to an embodiment of the present invention, the solvent for preparing the L-tartrate or L-tartrate crystal form (such as L-tartrate crystal form I) of Compound I is selected from one, two or three of methyl ethyl ketone, tetrahydrofuran, and ethyl acetate.

[0230] According to an embodiment of the present invention, the solvent for preparing the 1,5-naphthalenedisulfonate or 1,5-naphthalenedisulfonate crystal form of Compound I is selected from one, two or more of methanol, ethanol, isopropanol, methyl ethyl ketone, and water; for example, methanol, or a mixture of methyl ethyl ketone and water.

[0231] Exemplarily, the solvent for preparing 1,5-naphthalenedisulfonate crystal form I is one, two or three of methanol, ethanol, and isopropanol;

[0232] Exemplarily, the solvent for preparing 1,5-naphthalenedisulfonate crystal form II is a mixture of methyl ethyl ketone and water, preferably with a volume ratio of (10 - 25):1, such as 19:1.

[0233] According to an embodiment of the present invention, the mass-volume ratio of Compound I to the solvent is (15 - 150) mg:1 mL, for example (20 - 100) mg:1 mL, and exemplarily 30 mg:1.5 mL, 30 mg:0.6 mL, 20 mg:0.4 mL, 20 mg:0.5 mL, 250 mg:6 mL, 50 mg:0.5 mL, 50 mg:1 mL, 50 mg:1.75 mL, 500 mg:12 mL.

[0234] According to an embodiment of the present invention, the preparation method includes mixing and reacting a solution of Compound I with an acid solution to obtain the salt or crystal form of the salt of Compound I; or mixing and reacting a solution of Compound I with a solid acid to obtain the salt or crystal form of the salt of Compound I;

[0235] According to an embodiment of the present invention, the solvents in the solution of Compound I and the acid solution are the same.

[0236] According to an embodiment of the present invention, the preparation method includes a step of ultrasonic mixing.

[0237] According to an embodiment of the present invention, an acid solution is slowly added to a solution of Compound I.

[0238] According to an embodiment of the present invention, the temperature of the reaction is 20 - 35 °C, for example, 21 - 30 °C.

[0239] According to an embodiment of the present invention, the reaction is carried out under stirring or suspension milling.

[0240] According to an embodiment of the present invention, the reaction time is 10 hours - 10 days, for example, 15 hours - 6 days.

[0241] According to an embodiment of the present invention, the preparation method includes, after the reaction is completed, filtering to collect the solid product and drying.

[0242] The present invention also provides a pharmaceutical composition, which contains an active ingredient and optionally a pharmaceutically acceptable carrier, and the active ingredient is a salt of Compound I, a crystal form of the salt, or a free base crystal form.

[0243] For example, the pharmaceutically acceptable carrier includes, but is not limited to, one, two, or more of excipients, lubricants, binders, disintegrants, solvents, solubilizing agents, suspending agents, isotonic agents, buffers, preservatives, antioxidants, colorants, foaming agents, and flavoring agents (such as sweeteners, sour agents), etc.

[0244] Again, for example, the pharmaceutically acceptable carrier includes one, two, or more of water-soluble polymers, inorganic salts, etc.

[0245] According to an embodiment of the present invention, the pharmaceutical composition may further contain other active ingredients, such as other ROCK inhibitors.

[0246] The present invention also provides the use of the salt of Compound I, the crystal form of the salt, the free base crystal form, or the pharmaceutical composition in the preparation of a preparation.

[0247] The present invention also provides a preparation containing the salt of Compound I, the crystal form of the salt, or the free base crystal form.

[0248] According to an embodiment of the present invention, the preparation contains the above-mentioned pharmaceutical composition.

[0249] According to an embodiment of the present invention, the preparation may be in the form of powder, tablets (such as coated tablets, sustained-release or controlled-release tablets), lozenges, capsules (such as soft capsules or hard capsules), granules, pills, dispersible powders, suspensions, solutions, emulsions, elixirs, syrups, aerosols, creams, ointments, gels, injections, freeze-dried powder injections or suppositories and other dosage forms.

[0250] According to an embodiment of the present invention, the preparation may be administered in any of the following ways: oral administration, buccal administration, sublingual, inhalation, topical application, intravenous, subcutaneous, acupoint, intramuscular injection by parenteral administration, rectal administration.

[0251] According to an embodiment of the present invention, the preparation is a ROCK antagonist. Preferably, the ROCK antagonist is used for preventing and / or treating a disease caused by high expression of one or more ROCKs or overactivation of ROCK.

[0252] Preferably, the disease is selected from cardiovascular and cerebrovascular diseases, nervous system diseases, fibrosis diseases, eye diseases, tumors, arterial thrombosis disorders, radiation injuries, respiratory system diseases, autoimmune diseases, microbial infections, muscular dystrophy and diseases related to impaired lymphatic drainage, including atherosclerosis, acute coronary syndrome, hypertension, cerebral vasospasm, cerebral ischemia, ischemic stroke, restenosis, heart disease, heart failure, myocardial hypertrophy, myocardial ischemia-reperfusion injury, diabetes, diabetic nephropathy, cancer, neuronal degeneration, nerve injury diseases, spinal cord injury, erectile dysfunction, platelet aggregation, leukocyte aggregation, glaucoma, ocular hypertension, asthma, osteoporosis, pulmonary fibrosis (such as idiopathic interstitial pulmonary fibrosis), liver fibrosis, kidney fibrosis, COPD, renal dialysis, glomerulosclerosis, neuronal degenerative inflammation, fungal infection, bacterial infection, viral infection, Duchenne muscular dystrophy, fatty liver disease and fatty hepatitis.

[0253] The present invention also provides a method for preventing and / or treating a disease caused by high expression of ROCK or overactivation of ROCK, including administering a therapeutically effective amount of the salt or crystal form of the salt of Compound I, the free base crystal form, the pharmaceutical composition or the preparation to a subject.

[0254] Term Explanation

[0255] The term "crystal form" refers to a crystal form having the same chemical composition but different spatial arrangements of molecules and / or ions forming the crystal.

[0256] The term "amorphous" refers to a solid form of molecules and / or ions that is not crystalline. Amorphous solids do not show a definite X-ray powder diffraction pattern with distinct maxima.

[0257] The term "X-ray powder diffraction pattern substantially as shown" means that at least 50%, or at least 60%, or at least 70%, or at least 80%, or at least 90%, or at least 95%, or at least 99% of the main peaks shown in the X-ray powder diffraction pattern appear in the X-ray powder diffraction pattern; the main peaks refer to the peaks with a relative intensity greater than 10%, preferably greater than 20%, more preferably greater than 30% with the highest peak as a reference (the relative intensity of the highest peak is designated as 100%).

[0258] "Patient" and "subject" in the present invention have the same meaning and refer to a specific human or other warm-blooded mammal. The humans as "subjects" in the present invention include adults, infants, and children. Other warm-blooded mammals include but are not limited to non-human primates such as chimpanzees, other anthropoid apes or monkeys, as well as other zoo animals, domestic mammals or laboratory animals such as cats, pigs, dogs, cows, sheep, mice, rats, and guinea pigs, etc. Preferably, the "subject" in the present invention is a human.

[0259] Those skilled in the art can determine the appropriate amounts of any one, two, or any proportion of mixtures of salts, crystal forms of salts, or free base crystal forms in the pharmaceutical composition, as well as various pharmaceutically acceptable carriers and / or other active ingredients according to conventional methods.

[0260] The term "therapeutically effective amount" can be determined by a doctor with clinical practice qualifications in the art to be an amount of any one or two, or any proportion of mixtures of salts, crystal forms of salts, or free base crystal forms of the present invention, pharmaceutical composition or preparation sufficient to achieve the intended application (including but not limited to the treatment of diseases as defined above). Determining the therapeutically effective dose is within the ability of a clinician or researcher in the art and can be changed due to factors such as the intended application (in vitro or in vivo), or the subject and disease condition being treated, such as the weight and age of the subject, general health status, severity of the disease condition, route of administration, and other factors affecting the efficacy such as drug allergy history, etc. The specific dosage to be administered will vary depending on factors such as the specific compound or crystal form selected, the dosing regimen followed, whether co-administered with other compounds, the timing of administration, the tissue to which the drug is administered, and the physical delivery system carried.

[0261] The term "room temperature" refers to a temperature of 20 - 25 °C.

[0262] Those skilled in the art can understand that the "free base" or "free base of Compound I" in the present invention refers to Compound I (in the non-salted form).

[0263] Advantages of the present invention:

[0264] The inventors unexpectedly found that the free base crystal forms, salts, and crystal forms of salts of Compound I provided by the present invention have stable physical and chemical properties; the crystal forms of the present invention are relatively stable under high-temperature or high-humidity conditions.

[0265] Meanwhile, the free base crystal forms, salts, and crystal forms of salts of Compound I provided by the present invention have good in vitro and in vivo biological properties, which are more conducive to the quality control and drug-likeness of drugs. BRIEF DESCRIPTION OF THE DRAWINGS

[0266] Figure 1-1 is the XRPD characterization of free base crystal form I;

[0267] Figure 1-2 is the spectral analysis of the XRPD of free base crystal form I;

[0268] Figure 1-3 is the DSC-TGA spectrum of free base crystal form I;

[0269] Figure 1-4 is of free base crystal form I 1 1H-NMR spectrum;

[0270] Figure 2-1 is the XRPD characterization of hydrochloride crystal form I;

[0271] Figure 2-2 is the spectral analysis of the XRPD of hydrochloride crystal form I;

[0272] Figure 2-3 is the DSC-TGA spectrum of hydrochloride crystal form I;

[0273] Figure 2-4 is the polarized light micrograph of hydrochloride crystal form I;

[0274] Figure 2-5 is of hydrochloride crystal form I 1 1H-NMR spectrum;

[0275] Figure 3-1 is the XRPD characterization of sulfate crystal form I;

[0276] Figure 3-2 is the spectral analysis of the XRPD of sulfate crystal form I;

[0277] Figure 3-3 is the DSC-TGA spectrum of sulfate crystal form I;

[0278] Figure 3-4 is the polarized light micrograph of sulfate crystal form I;

[0279] Figure 3-5 is of sulfate crystal form I 1 1H-NMR spectrum;

[0280] Figure 4-1 XRPD characterization of p-toluenesulfonate polymorph I;

[0281] Figure 4-2 Spectrum analysis of XRPD of p-toluenesulfonate polymorph I;

[0282] Figure 4-3 DSC-TGA spectrum of p-toluenesulfonate polymorph I;

[0283] Figure 4-4 Polarizing micrograph of p-toluenesulfonate polymorph I;

[0284] Figure 4-5 For p-toluenesulfonate polymorph I 1 1H-NMR spectrum;

[0285] Figure 5-1 XRPD characterization of benzenesulfonate polymorph I;

[0286] Figure 5-2 Spectrum analysis of XRPD of benzenesulfonate polymorph I;

[0287] Figure 5-3 DSC-TGA spectrum of benzenesulfonate polymorph I;

[0288] Figure 5-4 Polarizing micrograph of benzenesulfonate polymorph I;

[0289] Figure 5-5 For benzenesulfonate polymorph I 1 1H-NMR spectrum;

[0290] Figure 6-1 XRPD characterization of maleate polymorph I;

[0291] Figure 6-2 Spectrum analysis of XRPD of maleate polymorph I;

[0292] Figure 6-3 DSC-TGA spectrum of maleate polymorph I;

[0293] Figure 6-4 Polarizing micrograph of maleate polymorph I;

[0294] Figure 6-5 For maleate polymorph I 1 1H-NMR spectrum;

[0295] Figure 6-6 XRPD overlay pattern of maleate polymorph I heated to 180 °C;

[0296] Figure 7-1 XRPD characterization of maleate polymorph II;

[0297] Figure 7-2 XRPD spectrum analysis of maleate polymorph II;

[0298] Figure 7-3 DSC-TGA spectrum of maleate polymorph II;

[0299] Figure 7-4 Polarized light micrograph of maleate polymorph II;

[0300] Figure 7-5 of maleate polymorph II 1 1H-NMR spectrum;

[0301] Figure 7-6 XRPD overlay of maleate polymorph II heated to 160 °C;

[0302] Figure 8-1 XRPD characterization of maleate polymorph III;

[0303] Figure 8-2 XRPD spectrum analysis of maleate polymorph III;

[0304] Figure 8-3 DSC-TGA spectrum of maleate polymorph III;

[0305] Figure 8-4 Polarized light micrograph of maleate polymorph III;

[0306] Figure 8-5 of maleate polymorph III 1 1H-NMR spectrum;

[0307] Figure 9-1 XRPD characterization of oxalate polymorph I;

[0308] Figure 9-2 XRPD spectrum analysis of oxalate polymorph I;

[0309] Figure 9-3 DSC-TGA spectrum of oxalate polymorph I;

[0310] Figure 9-4 Polarized light micrograph of oxalate polymorph I;

[0311] Figure 9-5 of oxalate polymorph I 1 1H-NMR spectrum;

[0312] Figure 10-1 XRPD characterization of camphorsulfonate polymorph I;

[0313] Figure 10-2DSC-TGA spectrum of camphorsulfonate polymorph I;

[0314] Figure 10-3 is of camphorsulfonate polymorph I 1 1H-NMR spectrum;

[0315] Figure 11-1 XRPD characterization of 2-hydroxyethanesulfonate polymorph I;

[0316] Figure 11-2 DSC-TGA spectrum of 2-hydroxyethanesulfonate polymorph I;

[0317] Figure 11-3 is of 2-hydroxyethanesulfonate polymorph I 1 1H-NMR spectrum;

[0318] Figure 12-1-1 XRPD characterization of ethanesulfonate polymorph I;

[0319] Figure 12-1-2 XRPD characterization of ethanesulfonate polymorph I prepared by scale-up;

[0320] Figure 12-2-1 DSC-TGA spectrum of ethanesulfonate polymorph I;

[0321] Figure 12-2-2 DSC-TGA spectrum of ethanesulfonate polymorph I prepared by scale-up;

[0322] Figure 12-3-1 is of ethanesulfonate polymorph I 1 1H-NMR spectrum;

[0323] Figure 12-3-2 is of ethanesulfonate polymorph I prepared by scale-up 1 1H-NMR spectrum;

[0324] Figure 12-3-3 is of ethanesulfonate polymorph I prepared by scale-up after methanol slurry 1 1H-NMR spectrum;

[0325] Figure 12-3-4 XRPD characterization of ethanesulfonate polymorph I prepared by scale-up before and after methanol slurry;

[0326] Figure 12-3-5 PLM image of ethanesulfonate polymorph I;

[0327] Figure 13-1 XRPD characterization of ethanesulfonate polymorph II;

[0328] Figure 13-2 DSC-TGA spectrum of ethanesulfonate polymorph II;

[0329] Figure 14-1 XRPD characterization of L-tartrate polymorph I;

[0330] Figure 14-2 DSC-TGA spectrum of L-tartrate polymorph I;

[0331] Figure 14-3 of L-tartrate polymorph I 1 1H-NMR spectrum;

[0332] Figure 15-1 XRPD characterization of 1,5-naphthalenedisulfonate polymorph I;

[0333] Figure 15-2 DSC-TGA spectrum of 1,5-naphthalenedisulfonate polymorph I;

[0334] Figure 15-3 of 1,5-naphthalenedisulfonate polymorph I 1 1H-NMR spectrum;

[0335] Figure 16-1 XRPD characterization of 1,5-naphthalenedisulfonate polymorph II;

[0336] Figure 16-2 DSC-TGA spectrum of 1,5-naphthalenedisulfonate polymorph II;

[0337] Figure 16-3 of 1,5-naphthalenedisulfonate polymorph II 1 1H-NMR spectrum;

[0338] Figure 17 DVS spectrum of free base polymorph I;

[0339] Figure 18 XRPD overlay of free base polymorph I before and after DVS measurement;

[0340] Figure 19 DVS spectrum of maleate polymorph III;

[0341] Figure 20 XRPD overlay of maleate polymorph III before and after DVS measurement;

[0342] Figure 21 DVS spectrum of ethylsulfonate polymorph I;

[0343] Figure 22 XRPD overlay of ethylsulfonate polymorph I before and after DVS measurement;

[0344] Figure 23 XRPD overlay of ethylsulfonate polymorph I before and after solubility measurement in biological medium FeSSIF; Detailed implementation manners

[0345] The technical solution of the present invention will be further described in detail below in combination with specific embodiments. It should be understood that the following embodiments are only used to illustrate and explain the present invention exemplarily, and should not be construed as limiting the protection scope of the present invention. All technologies implemented based on the above content of the present invention are covered within the scope of protection intended by the present invention.

[0346] Unless otherwise specified, the raw materials and reagents used in the following embodiments are all commercially available products, or can be prepared by known methods.

[0347] X-ray powder diffractometer (XRPD)

[0348] Using an X-ray powder diffractometer PANalytical Empyrean equipped with a PIXcel 1D detector to perform solid form analysis on the solids obtained in the experiment. The X-ray tube target material of the instrument is a copper target (K-Alpha( ))). The tube voltage and current are 45 KV and 40 mA respectively. The sample scanning range is from 3° 2θ to 40° 2θ, and the detection step size is 0.013° 2θ (except for tartrate crystal form and 1,5-naphthalenedisulfonate crystal form), and the detection step size for tartrate crystal form and 1,5-naphthalenedisulfonate crystal form is 0.0263° 2θ.

[0349] Differential scanning calorimetry (DSC)

[0350] The instrument used for DSC analysis is Discovery DSC 250 (TA Instruments, US) or TA Q2000 / 2500.

[0351] ① Using Discovery DSC 250 (TA Instruments, US) to perform thermal analysis on the sample: Weigh an appropriate amount of the sample and place it in the DSC sample pan and punch holes. After equilibrating the sample at 25 °C, heat it to the final temperature at a rate of 10 °C / min.

[0352] ② Collect DSC graphs on a TA Q2000 / 2500 differential scanning calorimeter, and the parameters are as follows:

[0353] Parameter DSC Method Linear heating rate Sample pan Aluminum pan, with / without crimping Temperature range 25℃ - Set end temperature Purge rate (℃ / min) 10 Protective gas Nitrogen

[0354] Thermogravimetric analysis (TGA)

[0355] Thermogravimetric analysis (TGA) uses a TGA 55 (TA Instruments, US) or TA Q5000 / 5500 thermogravimetric analyzer. Specifically:

[0356] ①Thermogravimetric analysis of the sample was carried out using a TGA 55 (TA Instruments, US): The sample was placed in a peeled closed aluminum sample pan. After the sample mass was automatically weighed in the TGA furnace, the sample was heated from room temperature to the final temperature at a rate of 10 °C / min.

[0357] ②TGA graphs were collected on a TA Q5000 / 5500 thermogravimetric analyzer with the following parameters:

[0358] Parameter TGA Method Linear heating rate Sample pan Aluminum pan, open Temperature range Room temperature - Set end temperature Purge rate (℃ / min) 10 Protective gas Nitrogen

[0359] TG-DSC thermal analyzer

[0360] Thermal analysis of the sample was carried out using a TG-DSC thermal analyzer STA449F3 (Netzsch, Germany). An appropriate amount of the sample was weighed and placed in the sample pan. After the sample was equilibrated at 25 °C, it was heated to the final temperature at a rate of 10 °C / min.

[0361] Polarizing light microscopy analysis (PLM)

[0362] The instrument used for PLM was a Polarizing Microscope ECLIPSE LV100POL (Nikon, JPN).

[0363] Proton nuclear magnetic resonance spectroscopy analysis ( 1 1H-NMR)

[0364] The 1H NMR information of the sample was 1 confirmed by 1H-NMR. 1 The instrument used for 1H-NMR analysis was a Bruker AVANCE III HD 300 / 400 equipped with a Sample Xpress60 automatic sample injection system.

[0365] Dynamic vapor sorption analysis (DVS)

[0366] The instrument used for dynamic vapor sorption analysis was a Vsorp (ProUmid GmbH&Co.KG, Germany).

[0367] The moisture sorption / desorption test of the sample was carried out using a Vsorp (ProUmid GmbH&Co.KG, Germany) moisture sorption analyzer. The sample was placed in the peeled sample pan, and the change in the sample mass with humidity at 25 °C (0 - 90% RH) was recorded. The specific DVS test parameters are as follows:

[0368] Equilibrium condition: 0.01% per / 45 min Cycle weighing time: 10 min Minimum time interval: 50 min Maximum time interval: 2.0h Equilibrium condition: 40℃ @ 0% RH (relative humidity) 6 h Sample measurement temperature: 25℃ Adsorption humidity: 0, 10, 20, 30, 40, 50, 60, 70, 80, 90% RH Desorption humidity: 80, 70, 60, 50, 40, 30, 20, 10, 0% RH

[0369] High performance liquid chromatography analysis (HPLC)

[0370] The instrument used for HPLC analysis is Agilent HPLC 1260 series or Shimadzu LC-20ADxr.

[0371] ① When the instrument used is Agilent HPLC 1260 series:

[0372] A) As shown in the HPLC method used for solubility test:

[0373]

[0374]

[0375] B) The HPLC method used for stability test is as follows:

[0376]

[0377] ② When the instrument used is Shimadzu LC-20ADxr:

[0378] A) The HPLC method used for solubility test is as follows:

[0379]

[0380]

[0381] B) The HPLC method used for stability investigation is as follows:

[0382]

[0383] Ion chromatography (IC)

[0384] The instrument used for IC analysis is Thermo ICS-6000. The method used for ion chromatography test is as follows:

[0385]

[0386]

[0387] Reagent list

[0388]

[0389] Example 1: Preparation and characterization of Compound I The preparation method of Compound I is as follows:

[0390]

[0391] 1. Preparation of ethyl 3-(3-nitropyridin-4-yl)-2-oxopropionate (I-01)

[0392] Under nitrogen protection, 4-methyl-3-nitropyridine (10.0 g, 72.0 mmol) was dissolved in diethyl oxalate (50 mL). DBU (12.7 g, 83.0 mmol) was added to the reaction solution, and the mixture was stirred at room temperature overnight. Ice water (30.0 g) was added to quench the reaction system, and the pH was adjusted to 4 - 5 with 1N dilute hydrochloric acid. The mixture was filtered, and the filter cake was washed with ethyl acetate (100 mL × 2). After drying in an oven, 18.0 g of a red solid was obtained, and the crude product was directly used in the next step. LC-MS [M + H] + = 239.1

[0393] 2. Preparation of ethyl 1H-pyrrolo[2,3-c]pyridine-2-carboxylate (I-02)

[0394] Under hydrogen protection, I-01 (17.0 g, 70.0 mmol) was dissolved in anhydrous dichloromethane (200 mL). Palladium / carbon (3.4 g, Pd mass fraction in palladium / carbon was 10%) was added to the reaction solution, and the mixture was stirred at room temperature overnight. The mixture was filtered, and the filtrate was concentrated under reduced pressure. The residue after concentration was separated by silica gel column chromatography (PE / EA = 1 / 1) to obtain 6.0 g of a yellow oily liquid. The overall yield of the two steps was 43.8%. LC-MS [M + H] + = 191.1

[0395] 3. Preparation of 1-(tert-butyl) 2-ethyl 1H-pyrrolo[2,3-c]pyridine-1,2-dicarboxylate (I-03)

[0396] Under nitrogen protection, I-02 (6.0 g, 31.5 mmol) was dissolved in anhydrous dichloromethane (50 mL). Boc anhydride (7.6 g, 34.7 mmol) and DMAP (384 mg, 3.2 mmol) were added to the reaction solution, and the mixture was stirred at room temperature overnight. The mixture was concentrated under reduced pressure, and the obtained crude product was separated by silica gel column chromatography (PE / EA = 3 / 1) to obtain 7.5 g of a yellow oily liquid. The yield was 82.4%, LC-MS [M + H] + = 291.0

[0397] 4. Preparation of ethyl 6-benzyl-4,5,6,7-tetrahydro-1H-pyrrolo[2,3-c]pyridine-2-carboxylate (I-04)

[0398] Under nitrogen protection, dissolve I-03 (7.2 g, 24.7 mmol) in anhydrous DMF (50 mL), add benzyl bromide (4.2 g, 24.7 mmol), react at 80 °C for 4 hours, concentrate under reduced pressure. Dissolve the concentrated solid in ethanol (50 mL), add sodium borohydride (934 mg, 24.7 mmol) to the reaction solution, and react at room temperature for 3 hours. Quench with water (100 mL), extract the resulting mixture with ethyl acetate (100 mL × 3), wash the combined organic phases with saturated brine (100 mL), dry over anhydrous sodium sulfate, filter, concentrate the filtrate, and separate the crude product by silica gel column chromatography (PE / EA = 3 / 1) to obtain 3.5 g of a yellow oily liquid with a yield of 49.8%. LC-MS [M+H] + = 285.1.

[0399] 5. Preparation of compound 6-(tert-butyl)-2-ethyl 1,4,5,7-tetrahydro-6H-pyrrolo[2,3-c]pyridine-2,6-dicarboxylate (I-05)

[0400] Dissolve I-04 (3.5 g, 12.3 mmol) in methanol (40 mL), add Boc anhydride (4.0 g, 18.5 mmol) and palladium / carbon (700 mg, Pd mass fraction in palladium / carbon is 10%), and react overnight at room temperature under a hydrogen atmosphere. Filter, concentrate the filtrate, and separate the residue by silica gel column chromatography (PE / EA = 5 / 1) to obtain 1.8 g of a yellow oily liquid with a yield of 50.0%. MS [M / 2+H] + = 295.1.

[0401] 6. Preparation of compound 6-(tert-butyl)-2-ethyl 1-methyl-1,4,5,7-tetrahydro-6H-pyrrolo[2,3-c]pyridine-2,6-dicarboxylate (I-06)

[0402] Under nitrogen protection, dissolve I-05 (1.8 g, 6.1 mmol) in tetrahydrofuran (20 mL), cool to 0 °C, add sodium hydride (364 mg, 9.1 mmol, 60%) to the reaction system, stir at this temperature for 30 minutes, dropwise add iodomethane (1.7 g, 12.2 mmol), and then react at room temperature for 2 hours. Quench the reaction with saturated ammonium chloride (20 mL), extract the resulting mixture with ethyl acetate (30 mL * 3), combine the organic phases, wash with saturated brine (50 mL), dry over anhydrous sodium sulfate, filter, concentrate the filtrate to obtain 1.8 g of a yellow oily liquid (crude product), LC-MS [M+H] + = 308.9.

[0403] 7. Preparation of compound 6-(tert-butoxycarbonyl)-1-methyl-4,5,6,7-tetrahydro-1H-pyrrolo[2,3-c]pyridine-2-carboxylic acid (I-07)

[0404] Dissolve I-06 (4.0 g, 12.9 mmol) in methanol (40 mL), add sodium hydroxide solution (38.7 mL, 1 N), and react at 50 °C for 2 hours. Adjust the pH to 5 - 6 with dilute hydrochloric acid (1 N), extract with ethyl acetate (50 mL × 3), wash the organic phase with saturated brine (50 mL), dry over anhydrous sodium sulfate, filter, and concentrate the filtrate under reduced pressure to obtain 3.2 g of a purple oily liquid. LC-MS [M+H] + = 281.1.

[0405] 8. Preparation of tert-butyl 2-(3,3-difluoroazetidine-1-carbonyl)-1-methyl-1,4,5,7-tetrahydro-6H-pyrrolo[2,3-c]pyridine-6-carboxylate (I-08)

[0406] Dissolve I-07 (2.4 g, 8.5 mmol) in anhydrous DMF (20 mL), add 3,3-difluoroazetidine hydrochloride (1.65 g, 12.8 mmol), HATU (4.9 g, 12.8 mmol), and DIEA (3.3 g, 25.6 mmol), and react at room temperature for 1 hour. Quench the reaction with water (30 mL), extract the resulting mixture with ethyl acetate (30 mL × 3), wash the combined organic phases with saturated brine (50 mL), dry over anhydrous sodium sulfate, filter, and purify the residue after concentrating the filtrate under reduced pressure by silica gel column chromatography (PE / EA = 3 / 1) to obtain 2.5 g of a yellow oily liquid. The yield is 83.3%, LC-MS [M+H] + = 355.8.

[0407] 9. Preparation of (3,3-difluoroazetidin-1-yl)(1-methyl-4,5,6,7-tetrahydro-1H-pyrrolo[2,3-c]pyridin-2-yl)methanone hydrochloride (I-09):

[0408] Dissolve I-08 (1.8 g, 5.8 mmol) in anhydrous ethanol (20 mL), add an ethanol solution of hydrochloric acid (5 mL, mass fraction of HCl 33%), and react at room temperature for 3 hours. Concentrate the reaction solution by rotary evaporation under reduced pressure to obtain 2.0 g of a yellow solid. LC-MS [M+H] + = 256.0.

[0409] 10. Preparation of tert-butyl 4-(4-nitrophenyl)-1H-pyrazole-1-carboxylate (I-10)

[0410] tert-Butyl 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole-1-carboxylate (58.8 g, 0.2 mol), 1-bromo-4-nitrobenzene (20 g, 0.1 mol), anhydrous potassium carbonate (41.5 g, 0.3 mol) and dichlorobis(1,1'-bis(diphenylphosphino)ferrocene)palladium(II) (7.3 g, 10 mmol) were added to a mixed solvent of 1,4-dioxane and water (1,4-dioxane: water = 30:1 (v:v), 207 mL), and the mixture was purged with nitrogen three times. The reaction mixture was stirred at 100 °C in an oil bath for 5 h. After the reaction was completed, the mixture was filtered under reduced pressure, and the residue obtained after concentration of the filtrate was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 4:1) to obtain 16 g of a yellow solid with a yield of 46%. MS (M+1-100) = 190.05, and the 1H NMR spectrum showed that it was product I-10.

[0411] 11. Preparation of tert-Butyl 4-(4-aminophenyl)-1H-pyrazole-1-carboxylate (I-11)

[0412] I-10 (16 g, 55.3 mmol) was dissolved in methanol (160 mL), and palladium / carbon (3.2 g, the mass fraction of Pd in palladium / carbon was 10%) was added. A hydrogen balloon was installed, and the mixture was purged with hydrogen. The reaction was carried out at 45 °C in a hydrogen atmosphere for 16 h. Celite was added, and the mixture was filtered under reduced pressure. The filtrate was concentrated, and the yellow crude product was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 3:1) to obtain 9.6 g of a light white solid with a yield of 60%. MS (M+1) = 260.15.

[0413] 12. Preparation of tert-Butyl 4-(4-((2-chloro-5-fluoropyrimidin-4-yl)amino)phenyl)-1H-pyrazole-1-carboxylate (I-12):

[0414] I-11 (1.8 g, 6.9 mmol) was dissolved in ethanol (20 mL), and 2,4-dichloro-5-fluoropyrimidine (2.3 g, 13.8 mmol) and DIEA (3.6 g, 27.6 mmol) were added successively. The reaction mixture was stirred at 40 °C for 2 h. After the reaction was completed, the mixture was cooled to room temperature, quenched with water (20 mL), and extracted with ethyl acetate (15 mL × 3). The organic phases were combined, washed with saturated brine (30 mL × 2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain a crude product. The crude product was purified by silica gel column chromatography (PE / EA = 3 / 1) to obtain 2.3 g of a brown solid product with a yield of 85.2%. LC-MS [M+H] + = 389.9.

[0415] 13. Preparation of (6-(4-((4-(1H-pyrazol-4-yl)phenyl)amino)-5-fluoropyrimidin-2-yl)-1-methyl-4,5,6,7-tetrahydro-1H-pyrrolo[2,3-c]pyridin-2-yl)(3,3-difluorazetidin-1-yl)methanone (Compound I):

[0416] I-12 (1.6 g, 12.3 mmol) was dissolved in n-butanol (16 mL). To the reaction system was added I-09 (784 mg, 3.1 mmol), DIEA (1.2 g, 3.1 mmol), and the mixture was stirred at 120 °C overnight. After the reaction was completed and cooled to room temperature, water (20 mL) was added to quench the reaction. The resulting mixture was extracted with ethyl acetate (20 mL × 3). The organic phases were combined, washed with saturated brine (20 mL × 3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain a crude product. The crude product was purified by silica gel column chromatography (DCM / MeOH = 20 / 1) and then lyophilized to obtain 600 mg of a white solid. LC-MS [M+H] + = 508.7.

[0417] 1 H NMR (400 MHz, DMSO-d 6 ) δ 12.90 (s, 1H), 9.34 (s, 1H), 8.16 (s, 1H), 8.06 (d, J = 3.7 Hz, 1H), 7.91 (s, 1H), 7.76 (d, J = 8.7 Hz, 2H), 7.59 (d, J = 8.7 Hz, 2H), 6.46 (s, 1H), 4.73 (s, 2H), 4.56 (s, 4H), 3.92 (t, J = 5.5 Hz, 2H), 3.74 (s, 3H), 2.55 (t, J = 5.2 Hz, 2H).

[0418] Characterization

[0419] An appropriate amount of the starting material, the free base of Compound I, was subjected to PLM, DSC (Discovery DSC 250 (TA Instruments, US)), TGA (TGA 55 (TA Instruments, US)), XRPD, and 1 H-NMR characterization. The results are shown in Table 1-1.

[0420] The XRPD pattern showed that the free base had a relatively high crystallinity and was named free base crystalline form I ( Figure 1-1 and 1-2 ). 1 The H-NMR results showed that the Compound I sample had 0.5% DCM and 0.7% THF solvent residues ( Figure 1-4)。DSC results showed that the sample had only one endothermic peak and the melting point was 271 °C, which was caused by the melting of the free base crystal form I; TGA showed that there was no obvious weight loss of the free base crystal form I before 150 °C ( Figure 1-3 )。

[0421] Table 1-1 Characterization results of starting material compound I

[0422]

[0423] Example 2: Hydrochloride

[0424] Weigh 30 mg of compound I at room temperature and place it in a sample bottle, then add 0.5 mL of MeOH. Dilute 54 μL of hydrochloric acid (12 M) in 1 mL of MeOH, and slowly add the above diluted hydrochloric acid solution (100 μL, 1.1 equivalents) to the suspension of compound I. Stir at room temperature overnight (about 21 °C, 16 h). Filter, collect the sample (filter cake) and dry it under vacuum at 40 °C for about 4 hours for XRPD characterization. The specific information and results are summarized in Table 2-1.

[0425] Table 2-1 Preparation of hydrochloride

[0426] Group Solvent HCl dosage (equivalent) Result 1 MeOH 1.1 Hydrochloride crystal form I

[0427] Note: "Equivalent" in the table is the molar ratio of acid to compound I.

[0428] One hydrochloride crystal form was obtained in this experiment, named hydrochloride crystal form I. Perform PLM, DSC (Discovery DSC250 (TA Instruments, US)), TGA (TGA 55 (TA Instruments, US)), XRPD and 1 1H-NMR characterization, and the relevant characterization results are shown in Table 2-2, Figure 2-1 to Figure 2-5 as shown.

[0429] Hydrochloride crystal form I is extremely small crystals (<10 μm, Figure 2-4 ). There is about 0.3% weight loss before 90 °C; there are two endothermic peaks on the DSC graph, the temperature is about 56 °C, and the broad endothermic peak may be caused by the dehydration of the hydrochloride, and the endothermic peak at about 241 °C is caused by the melting and decomposition of the hydrochloride. 1 1H-NMR analysis showed that there was no residual organic solvent in the sample. IC analysis showed that the sample contained about 1 equivalent of chloride ions, so the salt formation ratio was 1 / 1 (Table 2-3). Hydrochloride crystal form I is a water-free crystal form with medium crystallinity.

[0430] Table 2-2 Solid state characterization results of hydrochloride crystal form I

[0431]

[0432] Note: m / d: Melting with decomposition; RT = room temperature

[0433] Table 2-3 IC data of hydrochloride crystal form I

[0434]

[0435]

[0436] Example 3: Sulfate

[0437] Weigh 30 mg of Compound I at room temperature and place it in a sample bottle, then add 0.5 mL of MeOH. Dilute 18 μL of concentrated sulfuric acid (18 M) in 1 mL of MeOH, and slowly add 100 μL (0.55 equivalent) of the diluted sulfuric acid solution to the suspension of Compound I. Stir overnight at room temperature (about 21 °C, 16 h). Filter, collect the sample (filter cake) and dry it under vacuum at 40 °C for about 4 h, and perform XRPD characterization. The specific information and results are summarized in Table 3-1.

[0438] Table 3-1 Preparation of sulfate

[0439] Group Solvent <![CDATA[H 2 SO 4 Dosage (equivalent)]]> XRPD 1 MeOH 0.55 Sulfate crystal form I

[0440] Note: "Equivalent" in the table is the molar ratio of acid to Compound I.

[0441] One sulfate crystal form was obtained in this experiment, named sulfate crystal form I. Perform PLM, DSC (Discovery DSC 250 (TA Instruments, US)), TGA (TGA 55 (TA Instruments, US)), XRPD and 1 1H-NMR characterizations on sulfate crystal form I, and the relevant characterization results are shown in Table 3-2 Figure 3-1 to Figure 3-5 as follows.

[0442] Sulfate crystal form I is an irregular crystal ( Figure 3-4 ). There is about 3% weight loss before 125 °C, while 1 1H-NMR analysis shows that there is no residual organic solvent in the sample, and it is speculated that it is sulfate dehydration (about 1 equivalent of water). There are two endothermic peaks on the DSC curve. The broad endothermic peak at about 104 °C may be caused by sulfate dehydration, and the endothermic peak at about 163 °C is caused by melting with decomposition of sulfate. IC analysis shows that this sample contains ~0.5 equivalent of sulfate ions (Table 3-3), so the salt formation ratio is 0.5 / 1. It is speculated that sulfate crystal form I is a hydrate.

[0443] Table 3-2 Solid-state characterization results of sulfate crystal form I

[0444]

[0445] Note: RT = room temperature

[0446] Table 3-3 IC data of sulfate crystal form I

[0447]

[0448]

[0449] Example 4: p-toluenesulfonate

[0450] Weigh 30 mg of Compound I into a sample bottle at room temperature, then add 0.6 mL of the selected solvent MeOH, and then add solid p-toluenesulfonic acid (11.2 mg, 1.1 equivalents). The suspension is stirred overnight at room temperature (about 21 °C, 16 h). Filter, collect the sample (filter cake) and dry it under vacuum at 40 °C for about 4 h, and perform XRPD characterization. The specific information and results are summarized in Table 4-1.

[0451] Table 4-1 Preparation of p-toluenesulfonate

[0452]

[0453] Note: "Equivalent" in the table is the molar ratio of acid to Compound I.

[0454] In this experiment, a p-toluenesulfonate crystal form was identified and defined as p-toluenesulfonate crystal form I. Perform PLM, DSC (Discovery DSC 250 (TA Instruments, US)), TGA (TGA 55 (TA Instruments, US)), XRPD and 1 1H-NMR characterization, and the relevant characterization results are shown in Table 4-2, Figure 4-1 to Figure 4-5 as shown below.

[0455] p-Toluenesulfonate crystal form I is an irregular crystal ( Figure 4-3 ). There is about 2% weight loss before 110 °C. 1 1H-NMR analysis shows that there is a very small amount of organic solvent residue in the sample. Therefore, the weight loss in TGA is mainly attributed to the desorption of water. There are two endothermic peaks on the DSC curve. The broad endothermic peak at about 94 °C is attributed to dehydration, and the endothermic peak at about 198 °C is due to melting accompanied by decomposition. Therefore, it is speculated that p-toluenesulfonate crystal form I is a hydrate.

[0456] Table 4-2 Solid state characterization results of p-toluenesulfonate crystal form I

[0457]

[0458] Note: m / d: Melting with decomposition; RT = room temperature

[0459] Example 5: Benzenesulfonate

[0460] Weigh 30 mg of Compound I into a sample vial at room temperature, then add 0.6 mL of the selected solvent MeOH, and then add solid benzenesulfonic acid (10.3 mg, 1.1 equivalents). The suspension is stirred overnight at room temperature (~21 °C, 16 h). Filter, collect the sample (filter cake) and dry it under vacuum at 40 °C for about 4 h, and perform XRPD characterization. The specific information and results are summarized in Table 5-1.

[0461] Table 5-1 Preparation of Benzenesulfonate

[0462]

[0463] Note: "Equivalent" in the table is the molar ratio of acid to Compound I.

[0464] In this experiment, a crystalline salt form was identified and defined, named Benzenesulfonate Crystal Form I. Perform PLM, DSC (Discovery DSC 250 (TA Instruments, US)), TGA (TGA 55 (TA Instruments, US)), XRPD and 1 1H-NMR characterization on the Benzenesulfonate Crystal Form I sample. The relevant characterization results are shown in Table 5-2 Figure 5-1 to Figure 5-5 as follows.

[0465] Benzenesulfonate Crystal Form I is composed of tiny crystals ( Figure 5-4 ). There is about 3.9% weight loss before 100 °C. The weight loss in TGA is mainly attributed to the desorption of water. The broad endothermic peak at about 109 °C in the DSC curve is attributed to the desorption of water, and the endothermic peak at about 176 °C is attributed to melting with decomposition. Benzenesulfonate Crystal Form I is judged to be a hydrate.

[0466] Table 5-2 Solid State Characterization Results of Benzenesulfonate Crystal Form I

[0467]

[0468] Note: m / d: Melting with decomposition; RT = room temperature

[0469] Example 6: Maleate

[0470] Method 1:

[0471] At room temperature, 30 mg of Compound I was weighed and placed in a sample bottle respectively, then suspended in 0.6 mL of the corresponding solvent in Table 6-1, and then solid maleic acid (7.5 mg, 1.1 equivalents) was added. The suspension was stirred overnight at room temperature (about 21 °C, 16 h). Filtered, the sample (filter cake) was collected and vacuum dried at 40 °C for about 4 h, and XRPD characterization was carried out.

[0472] Two crystalline forms of maleate were found in this experiment, named maleate crystalline form I and maleate crystalline form II respectively. The specific information and results are summarized in Table 6-1.

[0473] Table 6-1 Preparation of Maleate

[0474]

[0475] Note: "Equivalent" in the table is the molar ratio of acid to Compound I.

[0476] Method 2:

[0477] At room temperature, 20 mg of Compound I was weighed and placed in a sample bottle respectively, then the corresponding solvents shown in Table 6-2 were added respectively. 5.03 mg of solid maleic acid was dissolved in 0.1 mL of the selected solvent (where IPAC directly added solid maleic acid), and slowly added dropwise to the suspension of the free base. Stirred overnight at room temperature (about 22 °C, 24 h). Filtered, the sample (filter cake) was collected and vacuum dried at 40 °C for about 4 h, and XRPD characterization was carried out.

[0478] Another new crystalline form of maleate was found in this experiment, named maleic acid crystalline form III. The specific information and results are summarized in Table 6-2.

[0479] Table 6-2 Preparation of Maleate

[0480]

[0481] Note: "Equivalent" in the table is the molar ratio of acid to Compound I.

[0482] The three crystalline forms of maleate obtained in the above experiment were characterized by PLM, DSC (Discovery DSC 250 (TA Instruments, US)), TGA (TGA 55 (TA Instruments, US)), XRPD and 1 1H-NMR, and the relevant characterization results are shown in Table 6-3, Figures 6-1 to 6-5 , Figures 7-1 to 7-5 , Figures 8-1 to 8-5 .

[0483] Maleate crystalline form I (Group 1) is a microcrystal. There is about 2.6% weight loss before 180 °C, 1The sample for H-NMR analysis contains approximately 2.9% MeOH. Therefore, the weight loss in TGA is due to the removal of MeOH. The endothermic peak with a peak temperature of approximately 198 °C in the DSC curve is caused by melting accompanied by decomposition. The XRPD patterns before and after heating the sample to 180 °C are inconsistent ( Figure 6-6 ), so maleate polymorph I is a MeOH solvate.

[0484] Maleate polymorph II (group 3) is in the form of aggregated particles. There is no obvious weight loss before 100 °C. 1 The sample for H-NMR analysis contains approximately 2.8% THF. Therefore, the weight loss in TGA is due to the desorption of THF. There are two endothermic peaks in the DSC curve. The broad endothermic peak with a peak temperature of approximately 48 °C is due to the desorption of surface water, and the endothermic peak with a peak temperature of approximately 190 °C is caused by melting accompanied by decomposition. The XRPD patterns before and after heating the sample to 160 °C are consistent ( Figure 7-6 ), and maleate polymorph II is an anhydrous crystal form.

[0485] Maleate polymorph III (group 4) is in the form of tiny crystals. There is no obvious weight loss before 100 °C. 1 The sample for H-NMR analysis contains approximately 0.5% acetone. The endothermic peak with a peak temperature of approximately 197 °C in the DSC curve is caused by melting accompanied by decomposition. Maleate polymorph III is an anhydrous crystal form.

[0486] Table 6-3 Solid-state characterization results of maleate

[0487]

[0488] Note: m / d: melting accompanied by decomposition; RT = room temperature

[0489] Example 7: Oxalate

[0490] Weigh 30 mg of Compound I into a sample vial at room temperature, then add 0.6 mL of the selected solvent MeOH, and then add oxalic acid solid (5.9 mg, 1.1 equivalents) to make a suspension. The suspension is stirred overnight at room temperature (about 21 °C, 16 h). Filter, collect the sample (filter cake) and dry it under vacuum at 40 °C for about 4 h, and perform XRPD characterization. The specific information and results are summarized in Table 7-1.

[0491] Table 7-1 Preparation of oxalate

[0492] Group Solvent Usage amount of oxalic acid (equivalent) XRPD 1 MeOH 1.1 Oxalate crystal form I

[0493] Note: "Equivalent" in the table is the molar ratio of acid to Compound I.

[0494] An oxalate crystal form was obtained from the experiment and named oxalate crystal form I. The oxalate sample was characterized by PLM, DSC (Discovery DSC 250 (TA Instruments, US)), TGA (TGA 55 (TA Instruments, US)), XRPD, and 1 1H-NMR, and the relevant characterization results are shown in Table 7-2, Figures 9-1 to 9-5 as follows.

[0495] Oxalate crystal form I is a microcrystal ( Figure 9-4 ). There is a weight loss of about 4.1% at 60-160 °C. 1 1H-NMR analysis of the sample shows 3.9% MeOH residue ( Figure 9-5 ), so the weight loss in TGA is attributed to the removal of MeOH; the broad endothermic peak with a peak temperature of about 127 °C in the DSC curve is attributed to the removal of MeOH ( Figure 9-3 ). The IC results show that the acid-base ratio of the sample is about 1 / 1 (Table 7-3). Oxalate crystal form I is determined to be a MeOH solvate.

[0496] Table 7-2 Solid state characterization results of oxalate crystal form I

[0497]

[0498] Table 7-3 IC data of oxalate crystal form I

[0499]

[0500] Example 8: Camphorsulfonate

[0501] At room temperature, 20 mg of Compound I was weighed into sample bottles respectively, and then the corresponding solvents (0.4 mL, 20V) in Table 8-1 were added respectively, and then camphorsulfonic acid (9.2 mg, 1.0 equivalent) was added. The suspension was stirred at room temperature for 5 days, centrifuged, the solid sample was collected, and dried in a blast oven at room temperature for 24 h, and XRPD characterization was carried out. The specific information and results are shown in Table 8-1, Table 1', and Figure 10-1 as follows:

[0502] Table 8-1 Preparation of camphorsulfonate

[0503]

[0504]

[0505] Note: "Equivalent" in the table is the molar ratio of acid to Compound I;

[0506] "V" in the table represents the ratio of the volume of the solvent in the salt-forming system in mL to the mass of Compound I in g.

[0507] XRPD Analysis of Camphorsulfonate Crystal Form I

[0508]

[0509] An experiment yielded a camphorsulfonate crystal form, named camphorsulfonate crystal form I. The camphorsulfonate sample was characterized by TGA (TG-DSC thermal analyzer), DSC (TG-DSC thermal analyzer), and 1 1H-NMR, and the relevant characterization results are as Figure 10-2 , Figure 10-3 shown.

[0510] TGA showed that camphorsulfonate crystal form I had a weight loss of approximately ~5% before 180 °C ( Figure 10-2 ). There were two endothermic peaks on the DSC. The broad endothermic peak at around 83 °C might be due to sample dehydration, and the endothermic peak at around 198 °C might be due to sample melting ( Figure 10-2 ). 1 1H NMR indicated that the salt-forming ratio of the free base to camphorsulfonic acid was 1 / 1, with no solvent residue ( Figure 10-3 ). Camphorsulfonate crystal form I is a hydrate, containing approximately 2 equivalents of water.

[0511] Example 9: 2-Hydroxyethanesulfonate

[0512] Weigh 20 mg of Compound I into a sample vial at room temperature, then add the selected solvent EtOH (0.4 mL, 20V), and then add 2-hydroxyethanesulfonic acid (5.8 mg, 85 wt%, 1.0 equivalent). The suspension was slurried at room temperature for 5 days, centrifuged, and the solid sample was collected and dried in a blast oven at room temperature for 24 h, followed by XRPD characterization. The specific information and results are shown in Table 9-1, Table 2', and Figure 11-1 shown as follows:

[0513] Table 9-1 Preparation of 2-Hydroxyethanesulfonate

[0514]

[0515] Note: "Equivalent" in the table refers to the molar ratio of acid to Compound I;

[0516] "V" in the table represents the ratio of the volume of the solvent in the salt-forming system in mL to the mass of Compound I in g.

[0517] Table 2' XRPD Analysis of 2-Hydroxyethanesulfonate Crystal Form I

[0518]

[0519] The XRPD test results of 2-hydroxyethanesulfonate are as Figure 11-1As shown, a 2-hydroxyethanesulfonate crystal form was obtained in ethanol in this experiment, named 2-hydroxyethanesulfonate crystal form I. The 2-hydroxyethanesulfonate crystal form I sample was characterized by TGA (TG-DSC thermal analyzer), DSC (TG-DSC thermal analyzer), 1 1H-NMR, and the relevant characterization results are as Figure 11-2 , Figure 11-3 shown.

[0520] TGA showed no weight loss before 150 °C and about 5.4% weight loss before 300 °C ( Figure 11-2 ). DSC had an exothermic peak at around 258 °C, which might be caused by the decomposition of the sample ( Figure 11-2 ). 1 1H NMR indicated that the salt formation ratio of the free base to hydroxyethanesulfonic acid was 1 / 1 and there was no solvent residue ( Figure 11-3 ).

[0521] Example 10: Ethanesulfonate

[0522] Example 10-1

[0523] At room temperature, 20 mg of Compound I was weighed and placed in a sample bottle respectively, then the corresponding solvents in Table 10-1 (0.4 mL, 20V) were added respectively, and then ethanesulfonic acid (4.33 mg, 1.0 equivalent) was added respectively. The suspension was mixed and slurried at room temperature for 5 days, centrifuged, the solid sample was collected, dried in a blast oven at room temperature for 24 h, and XRPD characterization was carried out. The specific information and results are shown in Table 10-1.

[0524] Table 10-1 Preparation of Ethanesulfonate

[0525]

[0526] Note: "Equivalent" in the table is the molar ratio of acid to Compound I;

[0527] "V" in the table represents the ratio of the volume of the salt formation system solvent in mL to the mass of Compound I in g.

[0528] In this experiment, 2 kinds of ethanesulfonate crystal forms were obtained, named ethanesulfonate crystal form I and ethanesulfonate crystal form II respectively. The 2 obtained ethanesulfonate crystal forms were characterized by XRPD, TGA (TG-DSC thermal analyzer), DSC (TG-DSC thermal analyzer), 1 1H-NMR, PLM, and the characterization results are as Figure 12-1-1 , 12-2-1 , 12-3-1, 12-3-5, 13-1, 13-2, Table 4' and Table 5' shown.

[0529] PLM shows that etanesulfonic acid crystal form I is small particle crystals (~10μm) with relatively high crystallinity. TGA shows that etanesulfonic acid crystal form I has a weight loss of about 1.4% before 230 °C, which is attributed to the removal of the solvent; DSC shows that etanesulfonic acid crystal form I has an exothermic peak at about 265 °C, which may be caused by the decomposition of the sample( Figure 12-2-1 ). The crystal form I of etanesulfonate 1 1H NMR shows that the salt formation ratio of the free base and hydroxyethanesulfonic acid is 1 / 1, and there is about 1.3 wt% ethanol residue( Figure 12-3-1 ).

[0530] Etanesulfonate crystal form II has a weight loss of about 2.3% before 150 °C; etanesulfonate crystal form II has two endothermic peaks and one exothermic peak. The endothermic peak at about 85 °C is a broad peak, which may be caused by the desolvation of the sample. The endothermic peak at about 177 °C may be caused by the melting of the sample. The exothermic peak at about 222 °C may be caused by the decomposition of the sample( Figure 13-2 ). It is speculated that etanesulfonate crystal form II is a solvate.

[0531] Table 4’ XRPD analysis of etanesulfonate crystal form I

[0532]

[0533] Table 5’ XRPD analysis of etanesulfonate crystal form II

[0534]

[0535] Example 10-2 Scale-up preparation of etanesulfonate crystal form I

[0536] Weigh about 200 mg of the free base and disperse it in 2 mL of anhydrous ethanol. Dissolve 43.3 mg (1.0 equivalent) of etanesulfonic acid in 2 mL of ethanol, and slowly add the ethanol solution of etanesulfonic acid dropwise to the suspension of the free base. Stir at room temperature for 3 days, filter, collect the filter cake, and vacuum dry the sample (filter cake) at 45 °C for about 24 hours. Perform XRPD, TGA (TG-DSC thermal analyzer), DSC (TG-DSC thermal analyzer) and 1 1H-NMR characterization on the dried sample. The test results are as shown in Figure 12-1-2 、 Figure 12-2-2 、 Figure 12-3-2 .

[0537] The XRPD test results (Table 6’) are consistent with the crystal form of etanesulfonate crystal form I (Table 4’). TGA shows a weight loss of about 1.3% before 230 °C. The endothermic peak of DSC at about 264 °C may be caused by the melting and decomposition of the sample, and the exothermic peak at about 269 °C may be caused by the decomposition of the sample. 11H NMR indicated that the salt formation ratio of the free base to 2-hydroxyethanesulfonic acid was 1 / 1, and there was 1.3 wt% ethanol residue. The sample was slurried with methanol at 50 °C for 24 h, and there was no residual ethanol solvent ( Figure 12-3-3 ), and the solid sample obtained by methanol slurrying was subjected to XRPD detection, and the crystal form remained unchanged ( Figure 12-3-4 ), and it was still crystal form I of the ethanesulfonate, so crystal form I of the ethanesulfonate is a crystal form without water.

[0538] Table 6’ XRPD analysis obtained from the scale-up experiment of crystal form I of ethanesulfonate

[0539] Example 11 L-tartrate

[0540] According to Table 11-1, compound I and L-tartaric acid were weighed into glass vials respectively, and then the corresponding solvents with the corresponding volumes were added to obtain a suspension. At temperature T 1 and stirred under suspension for time t 1 , the solid was separated by centrifugation and dried in vacuo at 50 °C to obtain a solid. The XRPD characterization results of the obtained solid are shown in Table 11-3. Compound I has poor solubility, and the tartrate was prepared by suspension slurrying, and in some experiments, the salt formation was incomplete and there was a mixed crystal form of the free base.

[0541] Table 11-1 Preparation parameters of L-tartrate

[0542]

[0543]

[0544] Note: "eq" in the table is the molar ratio of acid to compound I;

[0545] "V" in the table represents the ratio of the volume of the solvent in the salt formation system in mL to the mass of compound I in g.

[0546] One crystal form of L-tartrate was obtained in this experiment and named crystal form I of L-tartrate. The obtained L-tartrate sample (group 1) was characterized by XRPD, TGA (TA Q5000 / 5500 thermogravimetric analyzer), DSC (TA Q2000 / 2500 differential scanning calorimeter) and 1 1H NMR, and the relevant characterization results are summarized in Tables 11-2, 11-3 and Figure 14-1 、 14-2 、14-3.

[0547] Table 11-2 Solid state characterization results of crystal form I of L-tartrate

[0548]

[0549] The TGA results showed that when the L-tartrate polymorph I was heated to 150 °C, there was a weight loss of approximately 6.8%; the DSC results showed that the sample had an endothermic peak at around 207 °C (initial temperature). 1 1H NMR was measured in DMSO-d 6 and the results showed that the molar ratio of the residual solvent MEK to Compound I was 0.06 (corresponding to a weight loss of 0.7%), and the acid-base molar ratio of L-tartaric acid to Compound I was 1.0.

[0550] XRPD analysis of L-tartrate polymorph I in Group 1 of Table 11-3

[0551]

[0552]

[0553] Example 12 1,5-Naphthalenedisulfonate

[0554] Weigh approximately 20 mg of Compound I and an equimolar amount (1 equivalent) of 1,5-naphthalenedisulfonic acid into an HPLC vial, and add 0.5 mL of the corresponding solvent in Table 12-1 respectively to obtain a suspension. After stirring the suspension at room temperature for about 6 days, the solid was separated by centrifugation and dried under vacuum at 50 °C overnight. The XRPD characterization results of the obtained solid are shown in Table 12-1.

[0555] Table 12-1 Preparation of 1,5-Naphthalenedisulfonic Acid

[0556]

[0557] Note: "Equivalent" in the table is the molar ratio of the acid to Compound I.

[0558] Two polymorphs of 1,5-naphthalenedisulfonic acid were obtained in this experiment, named 1,5-naphthalenedisulfonate polymorph I and 1,5-naphthalenedisulfonate polymorph II respectively. The obtained 1,5-naphthalenedisulfonate samples were characterized by TGA (TA Q5000 / 5500 thermogravimetric analyzer), DSC (TA Q2000 / 2500 differential scanning calorimeter) and 1 1H NMR, and the relevant characterization results are summarized in Tables 12-2, 12-3, 12-4 and Figure 15-1 、 15-2 、15-3, 16-1, 16-2, 16-3.

[0559] Table 12-2 Solid State Characterization Results of 1,5-Naphthalenedisulfonate

[0560]

[0561] TGA results showed that when the 1,5-naphthalenedisulfonate polymorph I was heated to 150 °C, there was a weight loss of approximately 8.3%; DSC results showed that the 1,5-naphthalenedisulfonate polymorph I had two endothermic peaks at 84.0 °C and 119.6 °C (peak temperatures) Figure 15-2 ). 1 1H NMR was measured in DMSO-d6. The results showed that no residual solvent MeOH was detected, and the molar ratio of 1,5-naphthalenedisulfonic acid to Compound I was 0.9.

[0562] TGA results showed that when the 1,5-naphthalenedisulfonate polymorph II was heated to 180 °C, there was a weight loss of approximately 8.0%; DSC results showed that the sample had three endothermic peaks at 125.4 °C, 170.5 °C and 217.3 °C (peak temperatures) Figure 16-2 ). 1 1H NMR was measured in DMSO-d6. The results showed that the molar ratio of the solvent MEK to API was 0.06 (0.6 wt%), and the molar ratio of 1,5-naphthalenedisulfonic acid to Compound I was 0.9.

[0563] Table 12-3 XRPD analysis of 1,5-naphthalenedisulfonate polymorph I

[0564]

[0565] Table 12-4 XRPD analysis of 1,5-naphthalenedisulfonate polymorph II

[0566]

[0567] Example 13 Performance Test

[0568] 1. Hygroscopicity Test

[0569] 1.1 Hygroscopicity Test of Free Base Polymorph I, Maleate Polymorph III, and Ethanesulfonate Polymorph I

[0570] Using a Vsorp (ProUmid GmbH&Co.KG, Germany) moisture sorption analyzer, approximately 90 mg of free base polymorph I, 80 mg of maleate polymorph III (Group 8), and 80 mg of ethanesulfonate polymorph I (Group 1) were weighed into a tared DVS pan respectively for the hygroscopicity test and evaluation of the solid form.

[0571] DVS data showed that the free base polymorph I had a weight increase of 0.30 / 0.46% in the range of 0.0% RH to 80 / 90% RH, indicating that the free base polymorph I was slightly hygroscopic. The XRPD pattern of the free base polymorph I did not change before and after the DVS test, and the crystal form remained unchanged. The detailed characterization results are shown in Figure 17-18 .

[0572] DVS data showed that maleate polymorph III (group 8) had a weight increase of 0.29 / 0.37% in the range of 0.0% RH to 80 / 90% RH, indicating that maleate polymorph III was slightly hygroscopic. The XRPD patterns of maleate polymorph III before and after DVS testing remained consistent, and the crystal form remained unchanged. Detailed characterization results are shown in Figure 19-20 。

[0573] DVS test results showed that ethasylate polymorph I had a weight gain of about 0.3% in the range of 0.0% RH to 80% RH, indicating that ethasylate polymorph I was slightly hygroscopic; the XRPD patterns of ethasylate polymorph I before and after DVS testing did not change, and the crystal form remained unchanged. Detailed characterization results are shown in Figure 21-22 。

[0574] 2. Solubility tests

[0575] 2.1 Solubility tests of free base polymorph I and maleate polymorph III

[0576] At 37 °C, the solubilities of free base polymorph I and maleate polymorph III (group 8) in relevant biological media (SGF, FaSSIF, and FeSSIF) were tested.

[0577] Weigh 15 mg of the sample and disperse it in 3.0 mL of biorelevant medium. At 37 °C, shake it on a shaker at a speed of 800 rpm. Take out 1 mL of the dispersion for filtration at 0.5, 2, and 24 h, respectively. Test the solubility of the filtrate by Agilent HPLC 1260 series, test the pH value of the filtrate by a pH meter, and characterize the crystal form of the filter cake by XRPD.

[0578] In the three biorelevant media, the solubilities of free base polymorph I and maleate polymorph III were relatively low (<0.05 mg / mL). The crystal form of free base polymorph I remained unchanged in FaSSIF and FeSSIF for 24 h and underwent a crystal form transformation in SGF. The crystal form of maleate polymorph III underwent a crystal form transformation in FaSSIF, FeSSIF, and SGF for 24 h. The relevant characterization results are summarized in Table 13-1.

[0579] Table 13-1 Solubility results in biorelevant media

[0580]

[0581] 2.2 Solubility tests of tartrate polymorph I, ethasylate polymorph I, 2-hydroxyethasylate polymorph I, and sulfate polymorph I

[0582] The solubility of tartrate polymorph I, ethanesulfonate polymorph I, 2-hydroxyethanesulfonate polymorph I and sulfate polymorph I in relevant biological media (SGF, FaSSIF and FeSSIF) and pure water was tested at 37 °C.

[0583] Weigh 4 portions of tartrate polymorph I, ethanesulfonate polymorph I and 2-hydroxyethanesulfonate polymorph I in parallel, each portion being about 5 mg, and place them in 10 mL centrifuge tubes. Then add 5 mL of the corresponding biological medium solution to each tube, cover the tubes and mix evenly by suspension. Weigh 10 portions of 1 mg of sulfate polymorph I in parallel and place them in centrifuge tubes. Add 2 mL of the corresponding biological medium solution to each tube and mix evenly by suspension. Place the above centrifuge tubes in a thermostatic shaking incubator (temperature 37 °C, shaking frequency 100 times / min) and shake. Take about 1 mL of the dispersion and filter it at 1 h, 2 h, 4 h and 24 h respectively, and test the solubility and calculate the concentration of the filtrate by Shimadzu LC-20Adxr.

[0584] Among the three biological media, the solubility of tartrate polymorph I in SGF is the highest, reaching about 50 μg / mL, followed by its solubility in FeSSIF, with the solubility distribution ranging from 16 to 26 μg / mL in 0 - 24 h. While its solubility in FaSSIF is the lowest, with the solubility only being 1 - 2 μg / mL within 24 h.

[0585] The solubility of ethanesulfonate polymorph I in SGF is the highest, being about 100 μg / mL in the first 4 h, and the solubility at 24 h can also reach 72 μg / mL. Its solubility in FeSSIF is the second, with the solubility being 85 μg / mL in the first 1 h and only 12 μg / mL after 24 h. While its solubility in FaSSIF is the lowest, with the solubility only being 2 - 3 μg / mL within 24 h.

[0586] The change of the solubility of 2-hydroxyethanesulfonate polymorph I with time in SGF is not obvious, all being about 56 μg / mL. Its solubility in FeSSIF is the highest at 1 h, reaching 85 μg / mL, and only 28 μg / mL at 24 h. The solubility data in FaSSIF is close to that of ethanesulfonate, being 3 - 5 μg / mL.

[0587] The solubility of sulfate polymorph I in SGF at 24 h is 24.57 μg / mL, and it is not detected in other biological medium solutions and pure water.

[0588] In the solubility tests of tartrate polymorph I, esylate polymorph I, and 2-hydroxyesylate polymorph I in three biological media, there was no significant change in the pH of all solutions. However, the XRPD patterns of tartrate polymorph I, esylate polymorph I, and 2-hydroxyesylate polymorph I in the three biological media changed, and the crystallinity decreased to a certain extent; especially, esylate polymorph I was almost amorphous in FeSSIF. And the transformation from crystalline form to amorphous form or the decrease in crystallinity in biological media is beneficial to increasing the in vivo exposure of the active ingredient.

[0589] The superimposed XRPD patterns of esylate polymorph I before and after the solubility test in the biological medium FeSSIF are shown as Figure 23 follows.

[0590] The relevant characterization results are summarized in Table 13-2.

[0591] Table 13-2 Solubility results in biorelevant media

[0592]

[0593]

[0594] 3. Solid-state stability test

[0595] 3.1 Solid-state stability test of free base polymorph I, maleate polymorph III, and esylate polymorph I

[0596] Take 15 mg of free base polymorph I, maleate polymorph III, and esylate polymorph I and keep them under the conditions in Table 13-3. Take samples at 0 day, 1 week, 2 weeks, and 30 days, dissolve them in diluent to prepare a solution of about 1.0 mg / mL, and perform HPLC analysis of chemical stability by Agilent HPLC 1260 series. XRPD tests are performed on the solid samples after 2 weeks and 30 days to analyze physical stability.

[0597] The relevant characterization results are summarized in Table 13-3. The stability results show that free base polymorph I is physically and chemically stable for 2 weeks under the four conditions of 60 °C / closed, 60 °C / closed / N 2 , 40 °C / 75% RH (open), and RT / closed, but maleate polymorph III undergoes slight degradation. Esylate polymorph I is physically and chemically stable under the conditions of 40 °C / 75% RH (open) and 60 °C / closed.

[0598] Table 13-3 Stability evaluation results of free base polymorph I, maleate polymorph III, and esylate polymorph I Note: [1] "2 weeks" refers to the XRPD results after being placed for 2 weeks under the conditions of RT / closed, 40°C / 75% RH (open), 60°C / closed, 60°C / closed / N; [2] "30 days" refers to the XRPD results after being placed for 30 days under the conditions of 40°C / 75% RH (open) and 60°C / closed. 2 The XRPD results after being placed for 2 weeks under each condition; [2] "30 days" refers to the XRPD results after being placed for 30 days under the conditions of 40°C / 75% RH (open) and 60°C / closed.

[0599] 3.2 Stability Tests of Tartrate Crystal Form I, Ethanesulfonate Crystal Form I, and 2-Hydroxyethanesulfonate Crystal Form I

[0600] Weigh two portions of approximately 25 mg of tartrate crystal form I, ethanesulfonate crystal form I, and 2-hydroxyethanesulfonate crystal form I into 2 mL liquid phase vials. After sealing the vials with tin foil, place them separately under the conditions of temperature 40°C / humidity 75% or temperature 25°C / humidity 60% for about 3 months. Take samples at 0 month, 2 weeks, 1 month, and 3 months, dissolve them in the diluent to prepare a solution of approximately 1.0 mg / mL, and perform HPLC analysis of chemical stability using Shimadzu LC-20ADxr.

[0601] The relevant characterization results are summarized in Table 13-4. The stability results show that the chemical properties of tartrate crystal form I are basically stable under the two conditions of 25°C / 60% closed and protected from light, and 40°C / 75% closed and protected from light; the chemical properties of ethanesulfonate crystal form I and 2-hydroxyethanesulfonate crystal form I are basically stable under the condition of 40°C / 75% closed and protected from light.

[0602] Table 13-4 Stability Evaluation Results of Tartrate Crystal Form I, Ethanesulfonate Crystal Form I, and 2-Hydroxyethanesulfonate Crystal Form I

[0603]

[0604] Biological Activity Test of the Compound in Example 14

[0605] 1 In Vitro Kinase Activity Evaluation Experiment

[0606] 1) In Vitro ROCK2 Kinase Activity Evaluation

[0607] ROCK2 Activity Screening. The 96-well (Cisbio) time-resolved fluorescence assay was used to detect ROCK2 activity. The ROCK2 assay was run in the following assay buffer: 5 mM MgCl 2(Sigma), 1 mM DTT (Sigma) and 1× kinase buffer. Diluted with kinase buffer, first add 7.5 μL of ROCK2 kinase (Invitrogen, PV3759) to a 96-well microplate to make the final concentration 0.4 ng / μL, then add 0.25 μL of Compound I with 1% (v / v) DMSO content, and incubate at room temperature for 0.5 h. To initiate the reaction, after mixing ATP (Aladdin) and substrate STK-substrate 2-biotin with kinase buffer, add 7.5 μL of the mixture to the microplate to make the final concentrations 6.739 μM and 1 μM respectively, and continue to incubate at room temperature for 2 h. Mix 5 mL of detection buffer with STK antibody-Cryptate, then take an appropriate volume and mix it with an equal volume of streptavidin-XL665, and add 10 μL of this mixture to the microplate to terminate the reaction. After continuing to incubate for about 1 h, read the plate on a Molecular Devices Spectra Maxi3x multi-functional microplate reader. Among them, the kinase buffer, STK-substrate 2-biotin, detection buffer, STK antibody-Cryptate, and streptavidin-XL665 are all from HTRF KinEASE-STK kit (Cisbio, 1000 tests, 61GSTXLA).

[0608] 2) In vitro evaluation of ROCK1 kinase selectivity

[0609] The ROCK1 activity was detected by 96-well (Cisbio) time-resolved fluorescence assay. The ROCK1 assay was run in the following assay buffer: 5 mM MgCl 2(Sigma), 1 mM DTT (Sigma) and 1X kinase buffer. Diluted with kinase buffer, first add 7.5 μL of ROCK1 kinase (Invitrogen) to a 96-well microplate to make the final concentration 0.4 ng / μL, then add 0.25 μL of the compound with 1% DMSO content, and incubate at room temperature for 0.5 h. To initiate the reaction, also using kinase buffer, mix ATP (Aladdin) and substrate STK-substrate 2-biotin together, and add 7.5 μL of the mixture to the microplate to make the final concentrations 3.53 μM and 1 μM respectively, and continue to incubate at room temperature for 2 h. Mix 5 mL of detection buffer with STK antibody-Cryptate, then take an appropriate volume and mix it with an equal volume of streptavidin-XL665, and add 10 μL of the mixture to terminate the reaction. After continuing to incubate for about 1 h, read the plate on a Molecular Devices SpectraMax i3x multi-functional microplate reader. Among them, kinase buffer, STK-substrate 2-biotin, detection buffer, STK antibody-Cryptate, and streptavidin-XL665 are all from HTRF KinEASE-STK kit (Cisbio, 1000 tests, 61GSTXLA).

[0610] In vitro kinase activity evaluation experiments found that the IC 50 of compound I for ROCK2 is 36.48 nM, and the IC 50 for ROCK1 > 4.5 μM, showing excellent ROCK inhibitory activity; especially showing good selective inhibition of ROCK2 kinase.

[0611] 2 In vitro cytotoxicity test

[0612] The in vitro cytotoxicity test of compound I was determined by the CCK-8 method in HepG2 cells. Collect HepG2 cells (Beijing Na Biotechnology) in the logarithmic growth phase, adjust the cell suspension concentration, plate at 50,000 cells / well in a 96-well cell culture plate, place the cells in a 5%, 37 °C cell culture incubator overnight. After the cell confluence in the plate reaches 80 - 90%, change the medium and add the test compounds or solvent (DMSO) at each concentration gradient, and incubate in a 5%, 37 °C cell culture incubator for 48 hours. After the treatment, discard the medium in the plate, wash twice with PBS, add 100 μL of CCK-8 working solution (Beyotime Biotechnology) to each well, incubate at 37 °C in the dark for 1.5 hours, and detect the absorbance value at OD 450nm of each well, and analyze and calculate the CC 50 of each compound.

[0613] The results found that the CC 50 range of compound I is > 100 μM, showing good safety.

[0614] 3. Pharmacokinetic Study in Rats

[0615] After accurately weighing Compound I, 2-hydroxyethanesulfonate of Compound I, ethanesulfonate of Compound I (Polymorph I), tartrate of Compound I, and sulfate of Compound I, the required volume of solvent was added first, stirred for 5 min, and ultrasonicated at 25 °C until no large particles were visible to the naked eye, and then stirred for 30 min to obtain a homogeneous suspension for administration.

[0616] Male SPF-grade SD rats (Beijing Vital River Laboratory Animal Technology Co., Ltd.) were adaptively acclimated (without water and food restriction), and then Compound I, 2-hydroxyethanesulfonate of Compound I, ethanesulfonate of Compound I (Polymorph I), tartrate of Compound I, and sulfate of Compound I were administered respectively by single-dose gavage. Blood was collected from the orbital venous plexus of rats at 0.167, 0.5, 1, 2, 3, 4, 6, 9, 12, and 24 h after administration. The blood was centrifuged at 3500×g for 10 min at 4 °C, the supernatant was transferred, and the concentration of Compound I in plasma was detected by LC-MS / MS (AB Sciex TRIPLE QUAD 3500). The data were analyzed by WinNonlin (version 5.2.1 Pharsight, Mountain View, CA) using a non-compartmental model to obtain PK parameters. The experimental data are shown in Table 14-1, and the in vivo exposure of the salt forms of Compound I was significantly better than that of Compound I.

[0617] Table 14-1 PK Experiments of the Compounds of the Present Invention on Rats

[0618]

[0619] In the table: The dosing dose marked with * is calculated based on Compound I.

[0620] The above describes the embodiments of the present invention. However, the present invention is not limited to the above embodiments. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principle of the present invention shall be included within the protection scope of the present invention.

Claims

1. A salt of Compound I, characterized in that, the salt is an acid addition salt of Compound I and any one of the following acids: hydrochloric acid, sulfuric acid, p-toluenesulfonic acid, benzenesulfonic acid, maleic acid, tartaric acid, oxalic acid, ethanesulfonic acid, 2-hydroxyethanesulfonic acid, camphorsulfonic acid, 1,5-naphthalenedisulfonic acid; 2. The salt according to claim 1, characterized in that, the acid is sulfuric acid, L-tartaric acid, R-tartaric acid, ethanesulfonic acid or 2-hydroxyethanesulfonic acid.

3. The salt according to claim 1, characterized in that, the hydrochloride of Compound I is an acid addition salt of Compound I and hydrochloric acid, and the molar ratio of Compound I to hydrochloric acid is 1:(0.9 - 1.2).

4. The salt according to claim 1, characterized in that, the sulfate of Compound I is an acid addition salt of Compound I and sulfuric acid, and the molar ratio of Compound I to sulfuric acid is 1:(0.4 - 0.6).

5. The salt according to claim 1, characterized in that, the p-toluenesulfonate of Compound I is an acid addition salt of Compound I and p-toluenesulfonic acid, and the molar ratio of Compound I to p-toluenesulfonic acid is 1:(0.9 - 1.2).

6. The salt according to claim 1, characterized in that, the maleate of Compound I is an acid addition salt of Compound I and maleic acid, and the molar ratio of Compound I to maleic acid is 1:(0.9 - 1.2).

7. The salt according to claim 1, characterized in that, the oxalate of Compound I is an acid addition salt of Compound I and oxalic acid, and the molar ratio of Compound I to oxalic acid is 1:(0.9 - 1.2).

8. The salt according to claim 1, characterized in that, the camphorsulfonate of Compound I is an acid addition salt of Compound I and camphorsulfonic acid, and the molar ratio of Compound I to camphorsulfonic acid is 1:(0.9 - 1.2).

9. The salt according to claim 1, characterized in that, the 2-hydroxyethanesulfonate of Compound I is an acid addition salt of Compound I and 2-hydroxyethanesulfonic acid, and the molar ratio of Compound I to 2-hydroxyethanesulfonic acid is 1:(0.9 - 1.2).

10. The salt according to claim 1, characterized in that, the ethanesulfonate of Compound I is an acid addition salt of Compound I and ethanesulfonic acid, and the molar ratio of Compound I to ethanesulfonic acid is 1:(0.9 - 1.2).

11. The salt according to claim 1, characterized in that, the tartrate of Compound I is an acid addition salt of Compound I and tartaric acid; the molar ratio of Compound I to tartaric acid is 1:(0.9 - 1.2).

12. The salt according to claim 1, characterized in that, the 1,5-naphthalenedisulfonate of Compound I is an acid addition salt of Compound I and 1,5-naphthalenedisulfonic acid, and the molar ratio of Compound I to 1,5-naphthalenedisulfonic acid is 1:(0.8 - 1.1).

13. The salt according to claim 1, characterized in that, the benzenesulfonate of Compound I is an acid addition salt of Compound I and benzenesulfonic acid, and the molar ratio of Compound I to benzenesulfonic acid is 1:(0.9 - 1.2).

14. The salt according to claim 1, characterized in that, the salt of Compound I contains crystal water or does not contain crystal water.

15. The salt according to claim 1, characterized in that, The salt of Compound I is amorphous.

16. The crystal form of the salt of Compound I as claimed in Claim 1, wherein, the crystal form is hydrochloride crystal form I; the X-ray powder diffraction of the hydrochloride crystal form I represented by 2θ angle using Cu-Kα radiation has characteristic peaks at 8.0±0.2°, 8.3±0.2°, 13.1±0.2°, 18.2±0.2°, 18.7±0.2°, 22.6±0.2°, 23.0±0.2°, 24.4±0.2°, 30.3±0.2°.

17. The crystal form of the salt according to Claim 16, wherein, the hydrochloride crystal form I further has characteristic peaks at 15.8±0.2°, 16.2±0.2°, 21.5±0.2°.

18. The crystal form of the salt according to Claim 16, wherein, the hydrochloride crystal form I has an X-ray powder diffraction pattern substantially as shown in Figure 2-1; alternatively, the X-ray powder diffraction of the hydrochloride crystal form I represented by 2θ angle using Cu-Kα radiation is as shown in Figure 2-2, with an error range of ±0.2°.

19. The crystal form of the salt according to any one of Claims 16-18, wherein, the hydrochloride crystal form I is an anhydrate.

20. The crystal form of the salt of Compound I as claimed in Claim 1, wherein, the crystal form is sulfate crystal form I; the X-ray powder diffraction of the sulfate crystal form I represented by 2θ angle using Cu-Kα radiation has characteristic peaks at 5.3±0.2°, 8.4±0.2°, 11.0±0.2°, 13.7±0.2°, 16.2±0.2°, 18.0±0.2°, 19.3±0.2°.

21. The crystal form of the salt according to Claim 20, wherein, the sulfate crystal form I further has characteristic peaks at 10.6±0.2°, 14.5±0.2°, 17.0±0.2°, 18.8±0.2°, 19.6±0.2°, 22.3±0.2°, 24.9±0.2°.

22. The crystal form of the salt according to Claim 20, wherein, the sulfate crystal form I has an X-ray powder diffraction pattern substantially as shown in Figure 3-1; alternatively, the X-ray powder diffraction of the sulfate crystal form I represented by 2θ angle using Cu-Kα radiation is as shown in Figure 3-2, with an error range of ±0.2°.

23. The crystal form of the salt according to any one of Claims 20-22, wherein, the sulfate crystal form I is a hydrate.

24. The crystal form of the salt according to any one of Claims 20-22, wherein, the sulfate crystal form I has a DSC-TGA pattern substantially as shown in Figure 3-3; alternatively, the sulfate crystal form I loses 0.1-5.0% of its weight in the temperature range from room temperature to 125±2°C, and / or the sulfate crystal form I has two endothermic peaks, with peak temperatures of 104±5°C and 163±5°C respectively.

25. The crystal form of the salt of Compound I as claimed in Claim 1, wherein, the crystal form is p-toluenesulfonate crystal form I; The toluenesulfonate polymorph I uses Cu-Kα radiation, and the X-ray powder diffraction expressed in 2θ angles has characteristic peaks at 5.1±0.2°, 10.2±0.2°, 11.0±0.2°, 11.8±0.2°, 18.9±0.2°, 19.2±0.2°, 21.0±0.2°, 26.1±0.2°, 26.5±0.2°.

26. The crystal form of the salt according to claim 25, wherein, the toluenesulfonate polymorph I has an X-ray powder diffraction pattern substantially as shown in Figure 4-1; alternatively, the toluenesulfonate polymorph I uses Cu-Kα radiation, and the X-ray powder diffraction expressed in 2θ angles is as shown in Figure 4-2, with an error range of ±0.2°.

27. The crystal form of the salt according to any one of claims 25-26, wherein, the toluenesulfonate polymorph I is a hydrate.

28. The crystal form of the salt of compound I according to claim 1, wherein, the crystal form is benzenesulfonate polymorph I, and the X-ray powder diffraction expressed in 2θ angles of the benzenesulfonate polymorph I has characteristic peaks at 6.0±0.2°, 12.0±0.2°, 13.2±0.2°, 18.2±0.2°, 22.2±0.2°.

29. The crystal form of the salt according to claim 28, wherein, the benzenesulfonate polymorph I also has characteristic peaks at 8.7±0.2°, 17.4±0.2°, 18.6±0.2°, 19.4±0.2°, 20.3±0.2°, 24.2±0.2°, 30.5±0.2°.

30. The crystal form of the salt according to claim 28, wherein, the benzenesulfonate polymorph I has an X-ray powder diffraction pattern substantially as shown in Figure 5-1; alternatively, the benzenesulfonate polymorph I uses Cu-Kα radiation, and the X-ray powder diffraction expressed in 2θ angles is as shown in Figure 5-2, with an error range of ±0.2°.

31. The crystal form of the salt according to any one of claims 28-30, wherein, the benzenesulfonate polymorph I is a hydrate.

32. The crystal form of the salt of compound I according to claim 1, wherein, the crystal form is selected from maleate polymorph I, maleate polymorph II, maleate polymorph III; the maleate polymorph I uses Cu-Kα radiation, and the X-ray powder diffraction expressed in 2θ angles has characteristic peaks at 9.3±0.2°, 12.0±0.2°, 13.2±0.2°, 17.4±0.2°, 20.0±0.2°, 20.5±0.2°, 24.8±0.2°. The malate polymorph II, using Cu-Kα radiation, has characteristic peaks in the X-ray powder diffraction expressed in 2θ angles at 5.4±0.2°, 11.0±0.2°, 12.1±0.2°, 13.4±0.2°, 14.4±0.2°, 16.2±0.2°, 16.8±0.2°, 17.2±0.2°, 20.3±0.2°, 20.6±0.2°; The malate polymorph III, using Cu-Kα radiation, has characteristic peaks in the X-ray powder diffraction expressed in 2θ angles at 5.3±0.2°, 10.7±0.2°, 14.6±0.2°, 16.9±0.2°, 25.8±0.2°, 26.3±0.2°.

33. The crystal form of the salt according to claim 32, wherein, the malate polymorph II further has characteristic peaks at 7.2±0.2°, 17.8±0.2°, 25.0±0.2°; and / or, the malate polymorph III further has characteristic peaks at 12.4±0.2°, 16.1±0.2°, 19.2±0.2°, 20.2±0.2°.

34. The crystal form of the salt according to claim 32, wherein, the malate polymorph I has an X-ray powder diffraction pattern substantially as shown in Figure 6-1; or, the malate polymorph I, using Cu-Kα radiation, has an X-ray powder diffraction shown in Figure 6-2 in terms of 2θ angles, with an error range of ±0.2°.

35. The crystal form of the salt according to any one of claims 32-34, wherein, the malate polymorph I is a solvate.

36. The crystal form of the salt according to claim 32, wherein, the malate polymorph II has an X-ray powder diffraction pattern substantially as shown in Figure 7-1; or, the malate polymorph II, using Cu-Kα radiation, has an X-ray powder diffraction shown in Figure 7-2 in terms of 2θ angles, with an error range of ±0.2°.

37. The crystal form of the salt according to any one of claims 32-33 and 36, wherein, the malate polymorph II is an anhydrate.

38. The crystal form of the salt according to claim 32, wherein, the malate polymorph III has an X-ray powder diffraction pattern substantially as shown in Figure 8-1; or, the malate polymorph III, using Cu-Kα radiation, has an X-ray powder diffraction shown in Figure 8-2 in terms of 2θ angles, with an error range of ±0.2°.

39. The crystal form of the salt according to any one of claims 32-33 and 38, wherein, the malate polymorph III is an anhydrate; or, the malate polymorph III has a DSC-TGA pattern substantially as shown in Figure 8-3; or, the malate polymorph III has an endothermic peak at a peak temperature of 197±5°C.

40. The crystal form of the salt of the compound I according to claim 1, wherein, The crystal form is oxalate crystal form I. The X-ray powder diffraction of the oxalate crystal form I using Cu-Kα radiation has characteristic peaks at 7.6 ± 0.2°, 10.6 ± 0.2°, 11.2 ± 0.2°, 12.6 ± 0.2°, 15.2 ± 0.2°, 18.4 ± 0.2°, and 24.3 ± 0.2° in terms of 2θ angle.

41. The crystal form of the salt according to claim 40, wherein, the oxalate crystal form I has an X-ray powder diffraction pattern substantially as shown in Figure 9-1; alternatively, the X-ray powder diffraction of the oxalate crystal form I using Cu-Kα radiation is as shown in Figure 9-2 in terms of 2θ angle, with an error range of ±0.2°.

42. The crystal form of the salt according to any one of claims 40-41, wherein, the oxalate crystal form I is a solvate.

43. The crystal form of the salt of compound I according to claim 1, wherein, the crystal form is camphorsulfonate crystal form I; the X-ray powder diffraction of the camphorsulfonate crystal form I using Cu-Kα radiation has characteristic peaks at 3.6 ± 0.2°, 9.6 ± 0.2°, 12.6 ± 0.2°, 14.8 ± 0.2°, 16.5 ± 0.2°, 17.4 ± 0.2°, and 19.0 ± 0.2° in terms of 2θ angle.

44. The crystal form of the salt according to claim 43, wherein, the camphorsulfonate crystal form I also has characteristic peaks at 7.3 ± 0.2°, 20.2 ± 0.2°, 21.5 ± 0.2°, 21.8 ± 0.2°, 23.8 ± 0.2°, 25.4 ± 0.2°, and 28.1 ± 0.2°.

45. The crystal form of the salt according to claim 43, wherein, the camphorsulfonate crystal form I has an X-ray powder diffraction pattern substantially as shown in Figure 10-1; alternatively, the camphorsulfonate crystal form I has the following X-ray powder diffraction characteristic peaks in terms of 2θ angle, with an error range of ±0.2° 46. The crystal form of the salt according to any one of claims 43-45, wherein, the camphorsulfonate crystal form I is a hydrate.

47. The crystal form of the salt of compound I according to claim 1, wherein, the crystal form is 2-hydroxyethanesulfonate crystal form I. The X-ray powder diffraction of the 2-hydroxyethanesulfonate crystal form I using Cu-Kα radiation has characteristic peaks at 8.5 ± 0.2°, 8.9 ± 0.2°, 13.2 ± 0.2°, 16.0 ± 0.2°, 18.0 ± 0.2°, 18.8 ± 0.2°, and 25.4 ± 0.2°.

48. The crystal form of the salt according to claim 47, wherein, the 2-hydroxyethanesulfonate crystal form I also has characteristic peaks at 15.4 ± 0.2°, 20.7 ± 0.2°, 22.3 ± 0.2°, 22.6 ± 0.2°, 24.7 ± 0.2°, and 26.1 ± 0.2°.

49. The crystal form of the salt according to claim 47, wherein, The 2-hydroxyethanesulfonate polymorph I has an X-ray powder diffraction pattern substantially as shown in Figure 11-1; Alternatively, the 2-hydroxyethanesulfonate polymorph I has the following X-ray powder diffraction characteristic peaks expressed in 2θ angles, with an error range of ±0.2° 50. The polymorph of the salt according to any one of claims 47-49, characterized in that, the 2-hydroxyethanesulfonate polymorph I is an anhydrate; Alternatively, the 2-hydroxyethanesulfonate polymorph I has a DSC-TGA pattern substantially as shown in Figure 11-2.

51. The polymorph of the salt according to any one of claims 47-49, characterized in that, the 2-hydroxyethanesulfonate polymorph I loses 0 to 0.5% in weight in the temperature range from room temperature to 150 ± 2 °C, and loses 5.4 ± 0.2% in weight in the temperature range from room temperature to 300 ± 2 °C; and / or, the 2-hydroxyethanesulfonate polymorph I has an exothermic peak at a peak temperature of 258 ± 5 °C.

52. The polymorph of the salt of compound I according to claim 1, characterized in that, the polymorph is selected from ethanesulfonate polymorph I and ethanesulfonate polymorph II; the ethanesulfonate polymorph I, using Cu-Kα radiation, has characteristic peaks in X-ray powder diffraction expressed in 2θ angles at 10.4 ± 0.2°, 14.8 ± 0.2°, 16.9 ± 0.2°, 19.3 ± 0.2°, 21.4 ± 0.2°, 24.6 ± 0.2°; the ethanesulfonate polymorph II, using Cu-Kα radiation, has characteristic peaks in X-ray powder diffraction expressed in 2θ angles at 4.5 ± 0.2°, 9.0 ± 0.2°, 10.6 ± 0.2°, 17.5 ± 0.2°, 19.2 ± 0.2°.

53. The polymorph of the salt according to claim 52, characterized in that, the ethanesulfonate polymorph II also has characteristic peaks at 13.5 ± 0.2°, 14.7 ± 0.2°, 16.2 ± 0.2°, 18.0 ± 0.2°, 22.7 ± 0.2°, 25.2 ± 0.2°, 26.9 ± 0.2°.

54. The polymorph of the salt according to claim 53, characterized in that, the ethanesulfonate polymorph I also has characteristic peaks at 15.8 ± 0.2°, 17.4 ± 0.2°, 20.5 ± 0.2°, 23.0 ± 0.2°, 26.2 ± 0.2°.

55. The polymorph of the salt according to claim 52 or 54, characterized in that, the ethanesulfonate polymorph I has an X-ray powder diffraction pattern substantially as shown in Figure 12-1-1 or Figure 12-1-2; Alternatively, the ethanesulfonate polymorph I has the following X-ray powder diffraction characteristic peaks expressed in 2θ angles, with an error range of ±0.2° Alternatively, the ethanesulfonate polymorph I has the following X-ray powder diffraction characteristic peaks expressed in 2θ angles, with an error range of ±0.2° and / or, the ethanesulfonate polymorph I is an anhydrate; Alternatively, the ethanesulfonate polymorph I has a DSC-TGA pattern substantially as shown in Figure 12-2-1 or 12-2-2; Alternatively, the etilsulfonate polymorph I has a weight loss of 1.4 ± 0.4% before 230 ± 2 °C; Alternatively, the etilsulfonate polymorph I has an exothermic peak at a peak temperature of 265 ± 5 °C and an endothermic peak at a peak temperature of 260 ± 5 °C, and the peak temperature of the endothermic peak < the peak temperature of the exothermic peak.

56. The crystal form of the salt according to any one of claims 52-53, characterized in that, the etilsulfonate polymorph II has an X-ray powder diffraction pattern substantially as shown in Figure 13-1; Alternatively, the etilsulfonate polymorph II has the following X-ray powder diffraction characteristic peaks expressed in 2θ angles, with an error range of ±0.2° and / or, the etilsulfonate polymorph II is an organic solvent complex or hydrate; Alternatively, the etilsulfonate polymorph II has a DSC-TGA pattern substantially as shown in Figure 13-2; Alternatively, the etilsulfonate polymorph II has a weight loss of 2.3 ± 0.2% before 150 ± 2 °C; Alternatively, the etilsulfonate polymorph II has an endothermic peak at a peak temperature of 85 ± 5 °C, an endothermic peak at a peak temperature of 177 ± 5 °C, and an exothermic peak at a peak temperature of 222 ± 5 °C.

57. The crystal form of the salt of compound I according to claim 1, characterized in that, the crystal form is L-tartrate polymorph I; The L-tartrate polymorph I has characteristic peaks at 9.2 ± 0.2°, 11.6 ± 0.2°, 15.5 ± 0.2°, 18.7 ± 0.2°, 20.8 ± 0.2°, 21.1 ± 0.2° in the X-ray powder diffraction expressed in 2θ angles using Cu-Kα radiation.

58. The crystal form of the salt according to claim 57, characterized in that, the L-tartrate polymorph I also has characteristic peaks at 7.2 ± 0.2°, 7.7 ± 0.2°, 12.1 ± 0.2°, 14.5 ± 0.2°, 19.4 ± 0.2°, 22.6 ± 0.2°.

59. The crystal form of the salt according to any one of claims 57-58, characterized in that, the L-tartrate polymorph I has an X-ray powder diffraction pattern substantially as shown in Figure 14-1; Alternatively, the L-tartrate polymorph I has the following X-ray powder diffraction characteristic peaks expressed in 2θ angles, with an error range of ±0.2° and / or, the L-tartrate polymorph I is an anhydrate; Alternatively, the L-tartrate polymorph I has a DSC-TGA pattern substantially as shown in Figure 14-2; Alternatively, the L-tartrate polymorph I has a weight loss of 2-10% in the temperature range from room temperature to 150 ± 2 °C; Alternatively, the L-tartrate polymorph I has an endothermic peak at a peak temperature of 213 ± 5 °C.

60. The crystal form of the salt of compound I according to claim 1, characterized in that, the crystal form is selected from 1,5-naphthalenedisulfonate polymorph I and 1,5-naphthalenedisulfonate polymorph II; The 1,5-naphthalenedisulfonate polymorph I has characteristic peaks at 7.7 ± 0.2°, 15.4 ± 0.2°, 18.0 ± 0.2°, 23.5 ± 0.2°, 24.0 ± 0.2°, 24.8 ± 0.2° in the X-ray powder diffraction expressed in 2θ angle using Cu-Kα radiation; The 1,5-naphthalenedisulfonate polymorph II has characteristic peaks at 3.1 ± 0.2°, 3.9 ± 0.2°, 8.6 ± 0.2°, 10.2 ± 0.2°, 12.7 ± 0.2°, 13.9 ± 0.2°, 25.5 ± 0.2° in the X-ray powder diffraction expressed in 2θ angle using Cu-Kα radiation.

61. The crystal form of the salt according to claim 60, characterized in that, the 1,5-naphthalenedisulfonate polymorph I further has characteristic peaks at 8.0 ± 0.2°, 8.6 ± 0.2°, 10.9 ± 0.2°, 11.9 ± 0.2°, 16.8 ± 0.2°, 17.7 ± 0.2°, 19.8 ± 0.2°, 22.6 ± 0.2°; and / or, the 1,5-naphthalenedisulfonate polymorph II further has characteristic peaks at 8.4 ± 0.2°, 11.4 ± 0.2°, 14.4 ± 0.2°, 15.6 ± 0.2°, 20.5 ± 0.2°, 20.9 ± 0.2°, 22.0 ± 0.2°, 23.8 ± 0.2°.

62. The crystal form of the salt according to any one of claims 60 - 61, characterized in that, the 1,5-naphthalenedisulfonate polymorph I has an X-ray powder diffraction pattern substantially as shown in Figure 15-1; or, the 1,5-naphthalenedisulfonate polymorph I has the following X-ray powder diffraction characteristic peaks expressed in 2θ angle, with an error range of ±0.2° and / or, the 1,5-naphthalenedisulfonate polymorph I is a hydrate.

63. The crystal form of the salt according to any one of claims 60 - 61, characterized in that, the 1,5-naphthalenedisulfonate polymorph II has an X-ray powder diffraction pattern substantially as shown in Figure 16-1; or, the 1,5-naphthalenedisulfonate polymorph II has the following X-ray powder diffraction characteristic peaks expressed in 2θ angle, with an error range of ±0.2° and / or, the 1,5-naphthalenedisulfonate polymorph II is a hydrate.

64. The preparation method of the salt of compound I according to any one of claims 1 - 15, comprising mixing compound I with an acid and reacting in solvent 2 to obtain the salt of compound I; the solvent 2 is selected from one, two or more of organic solvent 2, water, and a mixed solvent of organic solvent 2 and water; the organic solvent 2 is selected from one, two or more of methanol, ethanol, isopropanol, butanol, acetone, butanone, ethyl acetate, isopropyl acetate, methyl tert-butyl ether, dichloromethane, acetonitrile, tetrahydrofuran, n-heptane, dimethyl sulfoxide, 2-methyltetrahydrofuran, chloroform.

65. The preparation method of the crystal form of the salt of compound I according to any one of claims 16 - 63, characterized in that, It includes mixing and reacting Compound I with an acid to obtain a crystalline form of the salt of Compound I; the reaction is carried out in Solvent 2; For preparing crystalline form I of the hydrochloride salt, sulfate salt, p-toluenesulfonate salt, benzenesulfonate salt, oxalate salt, 2-hydroxyethanesulfonate salt of Compound I, Solvent 2 is selected from one or two of methanol and ethanol; For preparing crystalline form I of the maleate salt of Compound I, Solvent 2 is selected from one or two of methanol and ethanol; For preparing crystalline form II of the maleate salt of Compound I, Solvent 2 is selected from one, two or three of tetrahydrofuran, isopropanol, acetonitrile, and isopropyl acetate; when Solvent 2 is selected as acetonitrile, a solution of Compound I is mixed and reacted with a solid acid; for preparing crystalline form III of the maleate salt of Compound I, Solvent 2 is selected from one, two or three of acetone, butanol, acetonitrile, and isopropyl acetate, and during the preparation, a solution of Compound I is mixed and reacted with an acid solution, and the solvents in the solution of Compound I and the acid solution are the same; For preparing crystalline form I of the camphorsulfonate salt of Compound I, Solvent 2 is selected from one, two or more of methanol, ethanol, acetonitrile, tetrahydrofuran, butanone, and isopropyl acetate; For preparing crystalline form I of the ethanesulfonate salt of Compound I, Solvent 2 is selected from one, two or more of methanol, ethanol, acetonitrile, and butanone; For preparing crystalline form II of the ethanesulfonate salt of Compound I, Solvent 2 is isopropyl acetate; For preparing crystalline form I of the L-tartrate salt of Compound I, Solvent 2 is selected from one, two or more of butanone, tetrahydrofuran, and ethyl acetate; For preparing crystalline form I of the 1,5-naphthalenedisulfonate salt of Compound I, Solvent 2 is selected from one or two of methanol and ethanol; For preparing crystalline form II of the 1,5-naphthalenedisulfonate salt of Compound I, Solvent 2 is a mixture of butanone and water.

66. According to the preparation method described in claim 65, it is characterized in that, the mass-volume ratio of Compound I to Solvent 2 is (15 - 150) mg:1 mL; and / or, the temperature of the reaction is 20 - 35 °C; and / or, the reaction is carried out under stirring or suspension milling; and / or, the reaction time is 10 hours - 10 days.

67. According to the preparation method described in claim 65 or 66, it is characterized in that, the preparation method includes, after the reaction is completed, filtering to collect the solid product and drying.

68. A pharmaceutical composition, it is characterized in that, the pharmaceutical composition contains an active ingredient and optionally a pharmaceutically acceptable carrier, and the active ingredient is the salt of Compound I described in any one of claims 1 - 15 or the crystalline form of the salt of Compound I described in any one of claims 16 - 63.

69. According to the pharmaceutical composition described in claim 68, it is characterized in that, the pharmaceutical composition further contains other ROCK inhibitors.

70. Use of the salt of Compound I described in any one of claims 1 - 15, the crystalline form of the salt of Compound I described in any one of claims 16 - 63, or the pharmaceutical composition described in any one of claims 68 - 69 in the preparation of a preparation, and the preparation is a ROCK antagonist.

71. The application according to claim 70, wherein, the ROCK antagonist is used for preventing and / or treating one or more diseases caused by high expression of ROCK or over-activation of ROCK.

72. The application according to claim 71, wherein, the diseases are selected from cardiovascular diseases, nervous system diseases, fibrotic diseases, eye diseases, tumors, arterial thrombosis disorders, radiation injuries, respiratory diseases, autoimmune diseases, microbial infections, muscular dystrophy, and diseases related to impaired lymphatic drainage.

73. The application according to claim 71, wherein, the diseases are selected from atherosclerosis, acute coronary syndrome, hypertension, cerebral vasospasm, cerebral ischemia, ischemic stroke, restenosis, heart disease, diabetes, diabetic nephropathy, cancer, neuronal degeneration, nerve injury diseases, spinal cord injury, erectile dysfunction, platelet aggregation, leukocyte aggregation, glaucoma, ocular hypertension, asthma, osteoporosis, pulmonary fibrosis, liver fibrosis, kidney fibrosis, COPD, renal dialysis, glomerulosclerosis, neuronal degenerative inflammation, fungal infection, bacterial infection, viral infection, Duchenne muscular dystrophy, and fatty liver disease.

74. The application according to claim 71, wherein, the diseases are selected from heart failure, myocardial hypertrophy, myocardial ischemia-reperfusion injury, and fatty liver.

75. The application according to claim 73, wherein, the pulmonary fibrosis is idiopathic pulmonary interstitial fibrosis.

76. A preparation containing a salt of compound I according to any one of claims 1-15, a crystal form of a salt of compound I according to any one of claims 16-63, or a pharmaceutical composition according to any one of claims 68-69.

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