Medicinal salt of quinoline amine compound, crystal form and preparation method of medicinal salt

Abstract

The invention relates to a pharmaceutical salt of a quinoline amine compound, a crystal form and a preparation method of the pharmaceutical salt. Specifically, the invention provides a pharmaceutically acceptable salt of 8-chloro-N-(2, 2-difluorobenzo [d] [1, 3] dioxacyclopentane-5-yl) quinoline-2-amine, a crystal form and a preparation method thereof, and the corresponding salt has good stability and can be better used for clinical treatment.

Description

A pharmaceutically acceptable salt, crystal form and preparation method of quinolineamine compound

[0001] This application claims the benefit of Chinese Patent Application No. 2022114917754, filed on November 25, 2022. This application incorporates the entirety of the aforementioned Chinese Patent Application. Technical Field

[0002] The present invention relates to the pharmaceutical field and relates to a pharmaceutically acceptable salt, a crystal form and a preparation method of a quinolineamine compound. Background Art

[0003] miR-124 is widely expressed in tissues throughout the body, with particular high expression in brain tissue. Studies have shown that overexpression of miR-124 can promote the quiescent transition of activated macrophages and microglia, thereby inhibiting the autoimmune disease encephalomyelitis. Furthermore, miR-124 can promote the transformation of macrophages to the M2 type, thereby exerting an anti-inflammatory effect. miR-124 also affects T cell differentiation, with miR-124-treated T cells showing decreased levels of IFN-γ and TNFα. Overexpression of miR-124 exerts an anti-inflammatory effect by downregulating STAT3 protein, thereby reducing the expression of the inflammatory cytokine IL-17 and inhibiting the differentiation of Th17 cells. These studies suggest that the development of a new type of small molecule drug that upregulates miR-124 could be used to effectively treat related inflammatory diseases.

[0004] Published related patent applications include WO2010143169A2, WO2015001518A1, WO2016009065A2, WO2017158201A1 and WO2020127843A1, etc.

[0005] WO2022247920 discloses a class of quinolineamine compounds that can upregulate miR-124, the structure of which is shown below:

[0006] Salt formation can improve certain undesirable physicochemical or biological properties of drugs. Developing salts with superior physicochemical or pharmaceutical properties compared to 8-chloro-N-(2,2-difluorobenzo[d][1,3]dioxolan-5-yl)quinolin-2-amine is of great significance. Given the importance of solid drug crystal forms and their stability in clinical treatment, in-depth research on the polymorphic forms of pharmaceutically acceptable salts of the compound 8-chloro-N-(2,2-difluorobenzo[d][1,3]dioxolan-5-yl)quinolin-2-amine is also of great significance for the development of drugs suitable for industrial production and with good biological activity.

[0007] Summary of the Invention

[0008] In one aspect, the present disclosure provides pharmaceutically acceptable salts of the compound 8-chloro-N-(2,2-difluorobenzo[d][1,3]dioxolan-5-yl)quinolin-2-amine.

[0009] In some embodiments, the pharmaceutically acceptable salt is selected from the group consisting of p-toluenesulfonate, phosphate, sulfate, maleate, hydrobromide, and hydrochloride.

[0010] In other embodiments, the chemical ratio of the compound 8-chloro-N-(2,2-difluorobenzo[d][1,3]dioxolan-5-yl)quinolin-2-amine to the acid molecule is 1:0.5 to 1:3, including 1:0.5, 1:1, 1:2 or 1:3.

[0011] In some embodiments, the chemical ratio of the compound 8-chloro-N-(2,2-difluorobenzo[d][1,3]dioxolan-5-yl)quinolin-2-amine to the acid molecule is 1:0.5 or 1:1.

[0012] On the other hand, the present disclosure also provides a method for preparing a pharmaceutically acceptable salt of the compound 8-chloro-N-(2,2-difluorobenzo[d][1,3]dioxolan-5-yl)quinolin-2-amine, comprising the step of forming a salt of the compound 8-chloro-N-(2,2-difluorobenzo[d][1,3]dioxolan-5-yl)quinolin-2-amine with an acid.

[0013] In some embodiments, the acid used in the salt-forming reaction is selected from p-toluenesulfonic acid, phosphoric acid, sulfuric acid, maleic acid, hydrobromide, and hydrochloric acid.

[0014] In some embodiments, the solvent used in the salt-forming reaction is selected from at least one of ethanol, acetonitrile, acetone, methanol, and water.

[0015] On the other hand, the present disclosure also provides a crystalline form I of the p-toluenesulfonate salt of the compound 8-chloro-N-(2,2-difluorobenzo[d][1,3]dioxolan-5-yl)quinolin-2-amine, which has an X-ray powder diffraction pattern expressed in terms of a diffraction angle of 2θ, with characteristic peaks at 6.616, 8.996, 14.120, 19.138 and 22.592.

[0016] In some embodiments, the p-toluenesulfonate crystalline form I has an X-ray powder diffraction pattern expressed as a diffraction angle 2θ, with characteristic peaks at 6.616, 8.996, 14.120, 17.120, 19.138, 20.615, 22.592 and 25.033.

[0017] In some embodiments, the p-toluenesulfonate salt form I has an X-ray powder diffraction pattern expressed as a diffraction angle 2θ, with characteristic peaks at 6.616, 8.996, 13.304, 14.120, 17.120, 19.138, 20.615, 21.289, 22.592, 25.033, 26.101 and 26.503.

[0018] In other embodiments, the p-toluenesulfonate salt crystalline form I has an X-ray powder diffraction pattern represented by a diffraction angle of 2θ as shown in FIG1 .

[0019] The present disclosure also provides a method for preparing a p-toluenesulfonate salt of the aforementioned compound in crystalline form I, comprising the steps of mixing the compound 8-chloro-N-(2,2-difluorobenzo[d][1,3]dioxolan-5-yl)quinolin-2-amine and a solvent (1), (b) adding p-toluenesulfonic acid, and crystallizing, wherein the solvent (1) is selected from at least one of ethanol, acetone, acetonitrile, methanol, and water.

[0020] On the other hand, the present disclosure also provides a phosphate crystal form I of the compound 8-chloro-N-(2,2-difluorobenzo[d][1,3]dioxolan-5-yl)quinolin-2-amine, which has an X-ray powder diffraction pattern expressed in terms of a diffraction angle of 2θ, with characteristic peaks at 6.511, 9.685, 12.660, 14.012 and 15.704.

[0021] In some embodiments, the phosphate crystal form I has an X-ray powder diffraction pattern represented by a diffraction angle of 2θ, with characteristic peaks at 6.511, 9.685, 12.660, 14.012, 15.704, 16.517, 21.487 and 22.209.

[0022] In some embodiments, the phosphate form I has an X-ray powder diffraction pattern expressed as a diffraction angle of 2θ, with characteristic peaks at 6.511, 9.685, 12.660, 14.012, 15.704, 16.517, 18.507, 21.487, 22.209, 24.493 and 25.596.

[0023] In other embodiments, the X-ray powder diffraction pattern of the phosphate crystal form I expressed in terms of a diffraction angle of 2θ is shown in FIG2 .

[0024] The present disclosure also provides a method for preparing the phosphate salt crystal form I of the aforementioned compound, comprising the steps of mixing the compound 8-chloro-N-(2,2-difluorobenzo[d][1,3]dioxolan-5-yl)quinolin-2-amine and a solvent (2), (b) adding phosphoric acid, and crystallizing, wherein the solvent (2) is selected from acetonitrile.

[0025] On the other hand, the present disclosure also provides a sulfate salt crystal form I of the compound 8-chloro-N-(2,2-difluorobenzo[d][1,3]dioxolan-5-yl)quinolin-2-amine, which has an X-ray powder diffraction pattern expressed in terms of a diffraction angle of 2θ, with characteristic peaks at 5.687, 11.448, 14.670, 17.315, 23.323 and 24.907.

[0026] In some embodiments, the sulfate salt crystalline form I has an X-ray powder diffraction pattern represented by a diffraction angle of 2θ, with characteristic peaks at 5.687, 9.540, 11.448, 14.670, 17.315, 18.906, 23.323 and 24.907.

[0027] In some embodiments, the sulfate salt crystalline form I has an X-ray powder diffraction pattern expressed as a diffraction angle 2θ, with characteristic peaks at 5.687, 9.540, 11.448, 12.137, 14.670, 17.315, 17.465, 18.906, 21.682, 23.323 and 24.907.

[0028] In other embodiments, the X-ray powder diffraction pattern of the sulfate salt crystal form I expressed in terms of a diffraction angle of 2θ is shown in FIG3 .

[0029] The present disclosure also provides a method for preparing the sulfate salt crystalline form I of the aforementioned compound, comprising the steps of mixing the compound 8-chloro-N-(2,2-difluorobenzo[d][1,3]dioxolan-5-yl)quinolin-2-amine and a solvent (3), (b) adding sulfuric acid, and crystallizing, wherein the solvent (3) is selected from ethanol or acetone.

[0030] On the other hand, the present disclosure also provides a sulfate salt crystalline form II of the compound 8-chloro-N-(2,2-difluorobenzo[d][1,3]dioxolan-5-yl)quinolin-2-amine, which has an X-ray powder diffraction pattern expressed in terms of a diffraction angle of 2θ, with characteristic peaks at 11.895, 13.102, 15.607, 16.658 and 17.967.

[0031] In some embodiments, the sulfate salt crystalline form II has an X-ray powder diffraction pattern expressed as a diffraction angle 2θ, with characteristic peaks at 6.518, 11.895, 13.102, 15.607, 16.181, 16.658, 17.967 and 24.329.

[0032] In some embodiments, the sulfate II crystalline form has an X-ray powder diffraction pattern expressed as a diffraction angle 2θ, with characteristic peaks at 6.518, 10.808, 11.291, 11.895, 13.102, 15.196, 15.607, 16.181, 16.658, 17.967 and 24.329.

[0033] In other embodiments, the X-ray powder diffraction pattern of the sulfate salt II crystal form expressed in terms of a diffraction angle of 2θ is shown in FIG4 .

[0034] The present disclosure also provides a method for preparing the sulfate salt II crystal form of the aforementioned compound, comprising the steps of mixing the compound 8-chloro-N-(2,2-difluorobenzo[d][1,3]dioxolan-5-yl)quinolin-2-amine and a solvent (4), (b) adding sulfuric acid, and crystallizing, wherein the solvent (4) is selected from acetonitrile.

[0035] On the other hand, the present disclosure also provides a sulfate salt III crystalline form of compound 8-chloro-N-(2,2-difluorobenzo[d][1,3]dioxolan-5-yl)quinolin-2-amine, which has an X-ray powder diffraction pattern expressed in terms of a diffraction angle of 2θ, with characteristic peaks at 13.502, 16.610, 17.858, 22.247 and 26.671.

[0036] In some embodiments, the sulfate III crystalline form has an X-ray powder diffraction pattern expressed as a diffraction angle 2θ, with characteristic peaks at 8.675, 13.502, 16.610, 17.858, 22.247, 25.379, 26.671 and 27.029.

[0037] In some embodiments, the sulfate III crystalline form has an X-ray powder diffraction pattern expressed as a diffraction angle 2θ, with characteristic peaks at 8.675, 13.502, 16.610, 17.449, 17.858, 22.247, 24.292, 25.379, 26.671 and 27.029.

[0038] In other embodiments, the X-ray powder diffraction pattern of the sulfate III crystal form expressed in terms of a diffraction angle of 2θ is shown in FIG5 .

[0039] The present disclosure also provides a method for preparing the sulfate salt III crystalline form of the aforementioned compound, comprising the steps of mixing the compound 8-chloro-N-(2,2-difluorobenzo[d][1,3]dioxolan-5-yl)quinolin-2-amine and a solvent (5), (b) adding sulfuric acid, and crystallizing, wherein the solvent (5) is selected from a mixed solution of water and methanol.

[0040] On the other hand, the present disclosure also provides a maleate salt form I of the compound 8-chloro-N-(2,2-difluorobenzo[d][1,3]dioxolan-5-yl)quinolin-2-amine, which has an X-ray powder diffraction pattern expressed in terms of a diffraction angle of 2θ, with characteristic peaks at 8.101, 12.810, 16.435, 26.186 and 27.616.

[0041] In some embodiments, the maleate salt crystalline form I has an X-ray powder diffraction pattern represented by a diffraction angle of 2θ, with characteristic peaks at 8.101, 12.810, 16.435, 20.667, 25.309, 26.186 and 27.616.

[0042] In some embodiments, the maleate salt crystalline form I has an X-ray powder diffraction pattern expressed as a diffraction angle 2θ, with characteristic peaks at 8.101, 12.810, 14.598, 16.435, 20.667, 21.474, 22.460, 25.309, 26.186 and 27.616.

[0043] In other embodiments, the maleate salt crystalline form I has an X-ray powder diffraction pattern represented by a diffraction angle of 2θ as shown in FIG6 .

[0044] The present disclosure also provides a method for preparing the maleate salt crystalline form I of the aforementioned compound, comprising the steps of mixing the compound 8-chloro-N-(2,2-difluorobenzo[d][1,3]dioxolan-5-yl)quinolin-2-amine and a solvent (6), (b) adding maleic acid, and crystallizing, wherein the solvent (6) is selected from ethanol.

[0045] On the other hand, the present disclosure also provides a maleate II crystalline form of the compound 8-chloro-N-(2,2-difluorobenzo[d][1,3]dioxolan-5-yl)quinolin-2-amine, which has an X-ray powder diffraction pattern expressed in terms of a diffraction angle of 2θ, with characteristic peaks at 7.817, 8.777, 11.663, 16.222 and 26.051.

[0046] In some embodiments, the maleate salt II crystalline form has an X-ray powder diffraction pattern expressed as a diffraction angle 2θ, with characteristic peaks at 7.817, 8.777, 11.663, 12.691, 16.222, 21.791 and 26.051.

[0047] In some embodiments, the maleate salt II crystalline form has an X-ray powder diffraction pattern expressed as a diffraction angle 2θ, with characteristic peaks at 7.817, 8.451, 8.777, 11.663, 12.691, 15.028, 16.222, 17.399, 21.791 and 26.051.

[0048] In other embodiments, the maleate salt II crystalline form has an X-ray powder diffraction pattern represented by a diffraction angle of 2θ as shown in FIG7 .

[0049] The present disclosure also provides a method for preparing a maleate salt II crystalline form of the aforementioned compound, comprising the steps of mixing the compound 8-chloro-N-(2,2-difluorobenzo[d][1,3]dioxolan-5-yl)quinolin-2-amine and a solvent (7), (b) adding maleic acid, and crystallizing, wherein the solvent (7) is selected from at least one of acetonitrile, ethanol, methanol, or water.

[0050] On the other hand, the present disclosure also provides a hydrobromide salt of compound 8-chloro-N-(2,2-difluorobenzo[d][1,3]dioxolan-5-yl)quinolin-2-amine in form I, which has an X-ray powder diffraction pattern expressed in terms of a diffraction angle of 2θ, with characteristic peaks at 6.077, 12.258, 22.769, 24.662 and 26.579.

[0051] In some embodiments, the hydrobromide salt form I has an X-ray powder diffraction pattern expressed as a diffraction angle 2θ, with characteristic peaks at 6.077, 12.258, 22.769, 24.662, 25.696, 26.579 and 34.105.

[0052] In some embodiments, the hydrobromide salt form I has an X-ray powder diffraction pattern expressed as a diffraction angle 2θ, with characteristic peaks at 6.077, 12.258, 16.589, 20.396, 22.769, 24.662, 25.218, 25.696, 26.579 and 34.105.

[0053] In other embodiments, the X-ray powder diffraction pattern of the hydrobromide salt form I expressed in terms of a diffraction angle of 2θ is shown in FIG8 .

[0054] The present disclosure also provides a method for preparing the hydrobromide salt crystalline form I of the aforementioned compound, comprising the steps of mixing the compound 8-chloro-N-(2,2-difluorobenzo[d][1,3]dioxolan-5-yl)quinolin-2-amine and a solvent (8), (b) adding hydrobromic acid, and crystallizing, wherein the solvent (8) is selected from at least one of ethanol, acetonitrile, or acetone.

[0055] On the other hand, the present disclosure also provides a hydrochloride salt form I of the compound 8-chloro-N-(2,2-difluorobenzo[d][1,3]dioxolan-5-yl)quinolin-2-amine, which has an X-ray powder diffraction pattern expressed in terms of a diffraction angle of 2θ, with characteristic peaks at 13.229, 14.520, 18.561, 19.925 and 23.830.

[0056] In some embodiments, the hydrochloride salt form I has an X-ray powder diffraction pattern expressed as a diffraction angle 2θ, with characteristic peaks at 13.229, 14.520, 18.561, 19.925, 23.830, 24.603 and 25.325.

[0057] In some embodiments, the hydrochloride salt form I has an X-ray powder diffraction pattern expressed as a diffraction angle 2θ, with characteristic peaks at 13.229, 14.520, 18.561, 19.925, 21.085, 22.588, 23.830, 24.603, 25.325 and 26.708.

[0058] In other embodiments, the X-ray powder diffraction pattern of the hydrochloride salt form I expressed in terms of a diffraction angle of 2θ is shown in FIG9 .

[0059] The present disclosure also provides a method for preparing the hydrochloride salt form I of the aforementioned compound, comprising the steps of mixing the compound 8-chloro-N-(2,2-difluorobenzo[d][1,3]dioxolan-5-yl)quinolin-2-amine and a solvent (9), (b) adding hydrochloric acid, and crystallizing, wherein the solvent (9) is selected from at least one of ethanol, acetonitrile, or acetone.

[0060] On the other hand, the present disclosure also provides a hydrochloride II crystal form of the compound 8-chloro-N-(2,2-difluorobenzo[d][1,3]dioxolan-5-yl)quinolin-2-amine, which has an X-ray powder diffraction pattern expressed in terms of a diffraction angle of 2θ, with characteristic peaks at 11.888, 13.593, 17.851, 23.977 and 26.309.

[0061] In some embodiments, the hydrochloride salt form II has an X-ray powder diffraction pattern expressed as a diffraction angle 2θ, with characteristic peaks at 11.888, 13.593, 17.851, 19.114, 23.977, 25.878 and 26.309.

[0062] In some embodiments, the hydrochloride salt form II has an X-ray powder diffraction pattern expressed as a diffraction angle 2θ, with characteristic peaks at 11.888, 13.593, 14.492, 17.851, 19.114, 20.367, 23.977, 25.878, 26.309 and 29.779.

[0063] In other embodiments, the X-ray powder diffraction pattern of the hydrochloride salt form II expressed in terms of a diffraction angle of 2θ is shown in FIG10 .

[0064] The present disclosure also provides a method for preparing the hydrochloride salt II crystal form of the aforementioned compound, comprising the steps of mixing the compound 8-chloro-N-(2,2-difluorobenzo[d][1,3]dioxolan-5-yl)quinolin-2-amine and a solvent (10), (b) adding hydrochloric acid, and crystallizing, wherein the solvent (10) is selected from at least one of ethanol, methanol, or water.

[0065] Furthermore, the present invention discloses an X-ray powder diffraction pattern of the aforementioned compound crystal form expressed in terms of a diffraction angle 2θ, wherein the error range of the 2θ angle is ±0.2.

[0066] In certain embodiments, the method for preparing the crystal form described in the present disclosure further comprises any one of the steps of stirring and dissolving or heating and dissolving, filtering, washing or drying.

[0067] In some embodiments, the crystallization includes but is not limited to stirring crystallization (dissolution crystallization, slurry crystallization) and volatile crystallization.

[0068] In some embodiments, the drying method includes but is not limited to forced air drying and vacuum drying. The drying temperature is generally 25°C to 100°C, preferably 30°C to 70°C, such as 40°C, 50°C or 60°C.

[0069] On the other hand, the present disclosure also provides a pharmaceutical composition comprising the aforementioned crystal form and a pharmaceutically acceptable excipient.

[0070] The present disclosure also provides a pharmaceutical composition prepared from the aforementioned crystal form and a pharmaceutically acceptable excipient.

[0071] The present disclosure also provides a method for preparing a pharmaceutical composition, comprising the step of mixing the aforementioned crystal form with a pharmaceutically acceptable excipient.

[0072] The present disclosure also provides use of the aforementioned crystal form or pharmaceutical composition in the preparation of a drug for regulating miRNA levels; preferably, the miRNA is miR-124.

[0073] The present disclosure also provides use of the aforementioned crystalline form or pharmaceutical composition in a medicine for treating and / or preventing a disease or condition selected from inflammation and cancer.

[0074] In some embodiments, the inflammation is inflammatory bowel disease. In some embodiments, the cancer is melanoma or breast cancer.

[0075] The "2θ or 2θ angle" mentioned in the present disclosure refers to the diffraction angle, θ is the Bragg angle, and the unit is ° or degree; the error range of each characteristic peak 2θ is ±0.20 (including the case where the number exceeding 1 decimal place is rounded off), specifically -0.20, -0.19, -0.18, -0.17, -0.16, -0.15, -0.14, -0.13, -0.12, -0.11, -0.10, -0.09, -0.08, -0.07, -0.06, -0.05, -0.04, -0.03, -0.02, -0.01, 0.00, 0.01, 0.02, 0.03, 0.04, 0.05, 0.06, 0.07, 0.08, 0.09, 0.10, 0.11, 0.12, 0.13, 0.14, 0.15, 0.16, 0.17, 0.18, 0.19, 0.20.

[0076] The "differential scanning calorimetry or DSC" described in this disclosure refers to measuring the temperature difference and heat flow difference between a sample and a reference object during the process of heating or maintaining the sample at a constant temperature to characterize all physical and chemical changes related to thermal effects and obtain phase change information of the sample.

[0077] The drying temperature in the present disclosure is generally 25°C-100°C, preferably 30°C-70°C, and can be dried under normal pressure or reduced pressure.

[0078] The "pharmaceutically acceptable excipients" described in this disclosure include, but are not limited to, any adjuvant, carrier, glidant, sweetener, diluent, preservative, dye / colorant, flavoring agent, surfactant, wetting agent, dispersant, suspending agent, stabilizer, isotonic agent or emulsifier approved by the U.S. Food and Drug Administration for use by humans or livestock animals.

[0079] The "beating" mentioned in the present disclosure refers to a purification method that utilizes the property that a substance has poor solubility in a solvent but impurities have good solubility in a solvent. Beating purification can remove color, change the crystal form or remove a small amount of impurities.

[0080] The crystalline forms disclosed herein include but are not limited to solvates of Compound 1, and the solvents include but are not limited to water. BRIEF DESCRIPTION OF THE DRAWINGS

[0081] Figure 1 is an XRPD diagram of p-toluenesulfonate Form I.

[0082] Figure 2 is an XRPD pattern of phosphate I crystal form.

[0083] Figure 3 is the XRPD pattern of sulfate salt Form I.

[0084] Figure 4 is the XRPD pattern of sulfate salt Form II.

[0085] Figure 5 is the XRPD pattern of sulfate III crystal form.

[0086] Figure 6 is the XRPD pattern of maleate salt Form I.

[0087] Figure 7 is an XRPD pattern of maleate II crystal form.

[0088] FIG8 is an XRPD pattern of hydrobromide salt Form I.

[0089] Figure 9 is the XRPD pattern of hydrochloride Form I.

[0090] Figure 10 is the XRPD pattern of hydrochloride salt form II. DETAILED DESCRIPTION

[0091] The present disclosure is further described in detail by the following examples and experimental examples. These examples and experimental examples are for illustrative purposes only and are not intended to limit the scope of the present disclosure.

[0092] Test conditions of the instruments used in the experiment:

[0093] The structures of the compounds were determined by nuclear magnetic resonance (NMR) and / or mass spectrometry (MS). -6 The unit of ppm is given. NMR measurements were performed using a Bruker AVANCE-400 NMR spectrometer. The solvents used were deuterated dimethyl sulfoxide (DMSO-d6), deuterated chloroform (CDCl3), and deuterated methanol (CD3OD), and tetramethylsilane (TMS) was used as the internal standard.

[0094] MS measurements were performed using an Agilent 1200 / 1290 DAD-6110 / 6120 Quadrupole MS liquid chromatography-mass spectrometer (manufacturer: Agilent, MS model: 6110 / 6120 Quadrupole MS), a Waters ACQuity UPLC-QD / SQD (manufacturer: Waters, MS model: Waters ACQuity Qda Detector / Waters SQ Detector), and a THERMO Ultimate 3000-Q Exactive (manufacturer: THERMO, MS model: THERMO Q 15 Exactive).

[0095] HPLC analysis was performed using an Agilent 1260DAD high pressure liquid chromatograph (Sunfire C18 150×4.6 mm column) and a Thermo U3000 high pressure liquid chromatograph (Gimini C18 150×4.6 mm column).

[0096] XRPD is X-ray powder diffraction detection: the measurement is carried out using a BRUKER D8 X-ray diffractometer, specific collection information: Cu anode (40kV, 40mA), radiation: monochromatic Cu-Ka radiation Scanning mode: θ / 2θ, scanning range: 3-48°.

[0097] DSC is differential scanning calorimetry: the measurement was performed using a METTLER TOLEDO DSC 3+ differential scanning calorimeter with a heating rate of 10°C / min, from 25 to 300°C, and a nitrogen purge rate of 50 mL / min.

[0098] TGA is thermogravimetric analysis: the test was performed using a METTLER TOLEDO TGA 2 thermogravimetric analyzer with a heating rate of 10°C / min. The specific temperature range was referred to the corresponding spectrum, and the nitrogen purge rate was 50 mL / min.

[0099] DVS stands for dynamic moisture sorption: using the Surface Measurement Systems instrument, humidity starts at 50% and the humidity range is 0%-95% with a step of 10%. The judgment standard is that the mass change of each gradient dM / dT is ≤ 0.002%, TMAX is 360min, and there are two cycles.

[0100] The known starting materials disclosed herein can be synthesized by methods known in the art, or can be purchased from ABCR GmbH & Co. KG, Acros Organics, Aldrich Chemical Company, Accela ChemBio Inc, Darui Chemicals, etc.

[0101] The reaction progress in the examples was monitored by thin layer chromatography (TLC). The developing solvent used in the reaction, the eluent system for column chromatography used to purify the compound, and the developing solvent system for thin layer chromatography included: A: dichloromethane / methanol system, B: n-hexane / ethyl acetate system. The volume ratio of the solvent was adjusted according to the polarity of the compound, and a small amount of alkaline or acidic reagents such as triethylamine and acetic acid could also be added for adjustment.

[0102] Example 1. Synthesis of 8-chloro-N-(2,2-difluorobenzo[d][1,3]dioxolan-5-yl)quin-2-amine (Refer to the preparation method of Example 1 in application No. WO2022247920)

[0103] 2,8-Dichloroquinoline 1a (100 mg, 0.51 mmol, Bidex Pharmaceuticals) and 5-amino-2,2-difluoro-1,3-benzo[1,3]dioxolane 1b (105 mg, 0.61 mmol, Shanghai Haohong Pharmaceuticals) were dissolved in isopropanol (1 mL) and heated to 90°C for 12 hours. The reaction mixture was filtered and purified by HPLC (Waters 2767-SQ Detecor2, elution system: 0.1% formic acid in water and acetonitrile, acetonitrile gradient: 65%-85%, flow rate: 30 mL / min) to obtain the title compound 1 (150 mg, 89.0% yield).

[0104] MS m / z(ESI):335.0[M+1].

[0105] 1 H NMR (500MHz, DMSO-d6) δ9.99(s,1H),8.88(d,1H),8.17(d,1H),7.81(dd,1H),7.77(dd,1H),7.50(dd,1H),7.39(d,1H),7.32(t,1H),7.14(d,1H).

[0106] Test Example 1. Upregulation of miR-124

[0107] 1. Experimental Materials and Instruments

[0108] 1. Dynabead Human T-Activator CD3 / CD28 for T Cell Expansion and Activation (Gibco, 11131D)

[0109] 2. Pan T Cell Isolation Kit, human (Miltenyi, 130-096-535)

[0110] 3. Human interleukin-2 (Human IL-2) (Peprotech, 200-02-100)

[0111] 4. Small RNA extraction kit (microRNA extraction kit) (Qiagen, 217004)

[0112] 5. miScript II RT Kit (Qiagen, 218161)

[0113] 6. miScript SYBR Green PCR Kit (Qiagen, 218073)

[0114] 7. Phosphate buffer PBS, pH 7.4 (Shanghai Yuanpei Biotechnology Co., Ltd., B320)

[0115] 8. Bovine serum albumin, BSA (Biyuntian, ST023)

[0116] 9.EDTA (0.5M), pH 8.0 (Invitrogen, AM9260G)

[0117] 10. LS Columns (Miltenyi, 130-042-401)

[0118] 11. 24-well cell culture plate (Corning, 3524)

[0119] 12. 96-well plate (Corning, 3788)

[0120] 13. Cell culture incubator (Thermo, Stericycle i160)

[0121] 14. Real-time fluorescence quantitative PCR instrument (Applied biosystem, QuantStudio6Flex)

[0122] 15.PCR instrument (Applied biosystem, ProFlex)

[0123] 16. 96-well clear PCR plate, 0.2 mL (Applied biosystems, N8010560)

[0124] 17. RPMI1640 culture medium (Gibco, 11875119)

[0125] 18. Fetal bovine serum, FBS (Gibco, 10099-141)

[0126] 19. Magnetic rack (Invitrogen, DynaMag TM -2)

[0127] 20. Six-well cell culture plate (Thermo, 150239)

[0128] 21. Spectrophotometer (IMPLEN, NP80)

[0129] 22. Magnetic bead separation rack (QuadroMACS Separator) (Miltenyi Biotec, 130-090-976)

[0130] 23.miR124-3P-F primer (customized by Genewise)

[0131] 24.hsa-U6 detection primer (Tian Gen, CD201-0145)

[0132] 2. Experimental steps

[0133] The effects of the compounds on miR-124 expression were examined in T cells activated with CD3 / CD28 antibodies. After treatment with the compounds, total RNA was extracted from the activated T cells, and the resulting reverse-transcribed cDNA was used as a template for quantification using SYBR Green fluorescent quantitative PCR using specific miR-124 primers.

[0134] Isolation of T cells: Purchased human peripheral blood mononuclear cells (PBMCs) were counted and filtered, washed once with separation buffer (PBS pH 7.4, containing 0.5% BSA and 2 mM EDTA), and the supernatant was discarded. The cells were separated at a rate of 1 × 10 7 Add 40 μL of buffer and 10 μL of pan T cell biotin-antibody cocktail to each cell, resuspend the pellet and mix well, then incubate in a refrigerator at 4°C for 5 minutes. 7 Add 30 μL of buffer and 20 μL of T cell separation magnetic beads (Pan T Cell MicroBeads Cocktail) to each cell, mix well and incubate at 4°C for 10 minutes. Rinse the LS column with 3 ml of cell separation buffer in advance, pass the above cell suspension through the column, and then wash the column three times with 1 ml of cell separation buffer. The outflowing cell fluid is collected in a 15 ml filter tube, which is the enriched T cells. Count the cells and calculate the number of cells to be 1×10 6 RPMI1640 medium (complete medium) containing 10% FBS and 40 U / mL IL-2 was added at a density of 10 cells / mL and stored on ice until use.

[0135] T cell activation: per 1×10 6Add 25 μL of activated magnetic beads to each cell, remove the corresponding T cell-activating CD3 / CD28 magnetic beads, and place them in a 1.5 mL filter tube. Oscillate on a shaker for approximately 30 seconds before aspirating. Wash the activated magnetic beads three times in the filter tube with culture medium at a volume ratio greater than 1:1. Remove all wash buffer from the final wash and resuspend the activated magnetic beads in complete culture medium equal to the starting volume. Add the washed activated magnetic beads to the cell suspension and mix thoroughly. Remove the six-well plate and add 3 mL of cells to each well. Incubate in a 37°C, 5% CO2 cell culture incubator for 2 days.

[0136] Compound treatment: The compound stock solution is 20mM, diluted to 200μM with DMSO, and then diluted 4-fold to 50μM (50×) with complete medium, mixed and set aside. DMSO diluted 4-fold (25% DMSO) is used as a negative control well. Activate T cells for two days, pipette the cells evenly, use a magnetic stand and install a 1.5mL filter tube, remove the activated magnetic beads, and collect the cell suspension. After counting the cells, filter at 300xg for 10 minutes, discard the supernatant, and resuspend the cells to 1.02×10 6 / mL, 980 μL of cell suspension and 20 μL of 50× compound were added to each 24-well plate, with a final concentration of 1 μM. The cells were cultured in a 37°C, 5% CO2 cell culture incubator for 3 days.

[0137] RNA extraction: Collect T cells by filtration, filter at 1500 rpm for 3 minutes, rinse once with PBS, and discard the supernatant after filtration. Use a small RNA extraction kit according to the manufacturer's instructions to extract total cellular RNA. Add 700 μL of Trizol cell lysis buffer to the cell pellet, pipette evenly, and let it stand at room temperature for 5 minutes. Add 140 μL of chloroform, vortex to mix, and let it stand at room temperature for 3 minutes. Filter the chloroform-cell lysis buffer mixture at 12,000 x g for 15 minutes at 4°C. Transfer the upper layer of solution to a new RNase-free filter tube, add 1.5 volumes of anhydrous ethanol, and pipette several times. Transfer the solution to an RNA adsorption column and filter at 8,000 x g for 15 seconds. Wash the filter column once with 700 μL of RWT solution, filter at 8,000 x g for 15 seconds, wash twice with 500 μL of RPE solution, and filter at 8,000 x g for 2 minutes. Place the adsorption column in a new 2 mL filter tube and filter at 12,000 x g for 1 minute to remove any residual wash solution. Place the adsorption column in a new 1.5 mL filter tube, add 30-50 μL of RNase-free water, and filter at 12,000 x g for 2 minutes. The collected solution is the RNA solution, and the RNA concentration is measured using a spectrophotometer. Store the RNA solution in a freezer at -80°C.

[0138] Reverse Transcription: Place the extracted RNA template on ice. Remove the small RNA reverse transcription kit and thaw some of the components (including 5× miScript HiSpec Buffer, 10× miScript Nucleic Acids Mix, and RNase-free water) at room temperature. Thaw the miScript Reverse Transcriptase mix on ice. Each 10 μL reaction consists of: 5× miScript HiSpec Buffer (2 μL), 10× miScript Nucleic Acids Mix (1 μL), miScript Reverse Transcriptase mix (1 μL), RNase-free water (2 μL), and RNA template (4 μL). Prepare the reaction on ice. Place the sample in a PCR instrument and set the following program: 37°C for 60 minutes; 95°C for 5 minutes; store at 4°C. The completed reaction is the cDNA sample.

[0139] Fluorescence quantitative PCR: SYBR green staining was used to detect the transcription level of miR-124. The transcription level of the housekeeping gene U6 was also detected as an internal control. Thaw all reagents required for the small RNA SYBR green PCR kit to room temperature. Dilute each cDNA sample template 10-fold and then 5-fold with RNase-free water. Prepare the reaction mixture according to Table 1 below and add the reaction mixture to a 96-well PCR plate. Seal the plate with a sealing film and filter. Perform the PCR reaction on a fluorescence quantitative PCR instrument according to the steps in Table 2.

[0140] Table 1 Fluorescence quantitative PCR reaction components

[0141] Table 2 Fluorescence quantitative PCR steps

[0142] Table 3 Fluorescence quantitative PCR detection primer list

[0143] Data analysis: According to the CT value calculated by the software, the ratio of miR-124 expression level to the internal reference U6 expression level of each sample was calculated, that is, ΔCT (test compound) = CT miRNA-124 (Test compound)-CT U6 (Test compound) The relative expression level was calculated by the following formula: relative expression level (test compound) = 2 (-[ΔCT(test compound)-ΔCT(DMSO)]).

[0144] Compound 1 upregulated miR-124 by 3.9 times (fold), and had good activity in promoting miR124 upregulation.

[0145] Example 2: Preparation of p-toluenesulfonate Crystal Form I

[0146] About 10 mg of compound 1 was weighed and dissolved in 0.15 mL of ethanol. A p-toluenesulfonic acid solution (2 mol / L, 16.4 μL) was added, and the mixture was stirred for crystallization. After centrifugation, the solid was vacuum dried to obtain the product.

[0147] The product was identified as p-toluenesulfonate crystalline form I by X-ray powder diffraction analysis. The XRPD spectrum is shown in FIG1 , and the positions of the characteristic peaks are shown in Table 4.

[0148] The DSC spectrum showed that the endothermic peak was 207.15℃.

[0149] The TGA spectrum showed that the weight loss was 0.22% at 30℃-150℃.

[0150] DVS testing showed that under normal storage conditions (i.e., 25°C, 60% RH), the sample experienced a moisture gain of approximately 0.06%. Under accelerated storage conditions (i.e., 70% RH), the moisture gain was approximately 0.09%. Under extreme conditions (i.e., 90% RH), the moisture gain was approximately 0.23%. Furthermore, subsequent DVS testing revealed no change in the crystal form.

[0151] Table 4

[0152] Example 3: Preparation of p-toluenesulfonate Form I

[0153] About 10 mg of compound 1 was weighed, added to 0.15 mL of 10% water / methanol, and p-toluenesulfonic acid solution (2 mol / L, 16.4 μL), stirred for crystallization, and the solid was vacuum dried after centrifugation to obtain the title product.

[0154] Example 4: Preparation of p-toluenesulfonate Crystal Form I

[0155] About 10 mg of compound 1 was weighed and dissolved in 0.15 mL of acetone. A p-toluenesulfonic acid solution (2 mol / L, 16.4 μL) was added, and the mixture was stirred for crystallization. After centrifugation, the solid was vacuum dried to obtain the title product.

[0156] Example 5: Preparation of p-toluenesulfonate Crystal Form I

[0157] About 10 mg of compound 1 was weighed and dissolved in 0.15 mL of acetonitrile. A p-toluenesulfonic acid solution (2 mol / L, 16.4 μL) was added, and the mixture was stirred for crystallization. After centrifugation, the solid was vacuum dried to obtain the title product.

[0158] Example 6: Preparation of Phosphate I Crystalline Form

[0159] About 10 mg of compound 1 was weighed and dissolved in 0.15 mL of acetonitrile. Phosphoric acid (2 mol / L, 16.4 μL) was added, stirred for crystallization, and the solid was vacuum dried after centrifugation to obtain the product.

[0160] The product was defined as phosphate I crystal form by X-ray powder diffraction analysis. The XRPD spectrum is shown in FIG2 , and the positions of its characteristic peaks are shown in Table 5.

[0161] The DSC spectrum showed that the endothermic peak was 164.15℃.

[0162] The TGA spectrum showed that the weight loss was 1.60% at 30℃-140℃.

[0163] Table 5

[0164] Example 7: Preparation of Sulfate Crystal Form I

[0165] About 10 mg of compound 1 was weighed and dissolved in 0.15 mL of ethanol. Sulfuric acid (2 mol / L, 16.4 μL) was added, stirred for crystallization, and the solid was vacuum dried after centrifugation to obtain the product.

[0166] The product was defined as sulfate crystal form I by X-ray powder diffraction analysis. The XRPD spectrum is shown in FIG3 , and the positions of its characteristic peaks are shown in Table 6.

[0167] The DSC spectrum showed that the endothermic peaks were 196.39°C and 205.37°C.

[0168] The TGA spectrum showed that the weight loss was 0.38% at 30℃-150℃.

[0169] Table 6

[0170] Example 8: Preparation of Sulfate Crystal Form I

[0171] About 10 mg of compound 1 was weighed and dissolved in 0.15 mL of acetone. Sulfuric acid (2 mol / L, 16.4 μL) was added and stirred for crystallization. After centrifugation, the solid was vacuum dried to obtain the title product.

[0172] Example 9: Preparation of Sulfate II Crystalline Form

[0173] About 10 mg of compound 1 was weighed and dissolved in 0.3 mL of acetonitrile. Sulfuric acid (2 mol / L, 16.4 μL) was added, stirred for crystallization, and the solid was vacuum dried after centrifugation to obtain the product.

[0174] The product was defined as sulfate II crystal form by X-ray powder diffraction analysis. The XRPD spectrum is shown in FIG4 , and the positions of its characteristic peaks are shown in Table 7.

[0175] The DSC spectrum showed that the endothermic peak was 212.61℃.

[0176] The TGA spectrum showed that the weight loss was 0.60% at 30℃-150℃.

[0177] DVS testing showed that under normal storage conditions (i.e., 25°C, 60% RH), the sample experienced a moisture absorption weight gain of approximately 1.56%. Under accelerated storage conditions (i.e., 70% RH), the moisture absorption weight gain was approximately 1.94%. Under extreme conditions (i.e., 90% RH), the moisture absorption weight gain was approximately 4.11%. Furthermore, subsequent DVS testing revealed no change in the crystal form.

[0178] Table 7

[0179] Example 10: Preparation of Sulfate III Crystalline Form

[0180] About 10 mg of compound 1 was weighed, added to 0.15 mL of 10% water / methanol, added with sulfuric acid (2 mol / L, 16.4 μL), stirred for crystallization, centrifuged, and the solid was vacuum dried to obtain the product.

[0181] The product was defined as sulfate III crystal form by X-ray powder diffraction analysis. The XRPD spectrum is shown in FIG5 , and the positions of its characteristic peaks are shown in Table 8.

[0182] The DSC spectrum showed that the endothermic peak was 123.93℃.

[0183] The TGA spectrum showed that the weight loss was 2.39% at 30℃-100℃.

[0184] DVS testing showed that under normal storage conditions (i.e., 25°C, 60% RH), the sample experienced a moisture gain of approximately 0.66%. Under accelerated storage conditions (i.e., 70% RH), the moisture gain was approximately 0.76%. Under extreme conditions (i.e., 90% RH), the moisture gain was approximately 1.45%. Furthermore, subsequent DVS testing revealed no change in the crystal form.

[0185] Table 8

[0186] Example 11: Preparation of Maleate Salt Form I

[0187] About 10 mg of compound 1 was weighed and dissolved in 0.15 mL of ethanol. Maleic acid solution (2 mol / L, 16.4 μL) was added and stirred for crystallization. After centrifugation, the solid was vacuum dried to obtain the product.

[0188] The product was defined as maleate salt form I by X-ray powder diffraction analysis. The XRPD spectrum is shown in FIG6 , and the positions of the characteristic peaks are shown in Table 9.

[0189] The DSC spectrum showed that the endothermic peak was 162.42℃.

[0190] The TGA spectrum showed that the weight loss was 0.62% at 30℃-130℃.

[0191] Table 9

[0192] Example 12: Preparation of Maleate Salt Form II

[0193] About 10 mg of compound 1 was weighed and dissolved in 0.15 mL of acetonitrile. Maleic acid solution (2 mol / L, 16.4 μL) was added and stirred for crystallization. After centrifugation, the solid was vacuum dried to obtain the product.

[0194] The product was defined as maleate II crystal form by X-ray powder diffraction analysis. The XRPD spectrum is shown in FIG7 , and the positions of its characteristic peaks are shown in Table 10.

[0195] The DSC spectrum showed that the endothermic peak was 164.59℃.

[0196] The TGA spectrum showed that the weight loss was 1.61% from 30°C to 145°C.

[0197] DVS testing showed that under normal storage conditions (i.e., 25°C, 60% RH), the sample experienced a moisture gain of approximately 0.09%; under accelerated storage conditions (i.e., 70% RH), the moisture gain was approximately 0.11%; and under extreme conditions (i.e., 90% RH), the moisture gain was approximately 1.11%. Furthermore, subsequent DVS testing revealed no change in the crystal form.

[0198] Table 10

[0199] Example 13: Preparation of Maleate Salt Form II

[0200] About 10 mg of compound 1 was weighed, added to 0.15 mL of 10% water / methanol, and maleic acid solution (2 mol / L, 16.4 μL) was added, stirred for crystallization, centrifuged, and the solid was vacuum dried to obtain the title product.

[0201] Example 14: Preparation of Maleate Salt Form II

[0202] About 300 mg of compound 1 was weighed and dissolved in 4 mL of ethanol. Maleic acid solution (2 mol / L, 471 μL) was added and stirred for crystallization. After centrifugation, the solid was vacuum dried to obtain the title product.

[0203] Example 15: Preparation of Maleate Salt Form II

[0204] About 10 mg of compound 1 was weighed and dissolved in 0.15 mL of ethanol. Maleic acid solution (2 mol / L, 16.4 μL) was added and stirred for crystallization. After centrifugation, the solid was vacuum dried to obtain the title product.

[0205] Example 16: Preparation of Hydrobromide Crystal Form I

[0206] About 10 mg of compound 1 was weighed and dissolved in 0.15 mL of ethanol. Hydrobromic acid (2 mol / L, 16.4 μL) was added and stirred for crystallization. After centrifugation, the solid was vacuum dried to obtain the product.

[0207] The product was defined as hydrobromide salt form I by X-ray powder diffraction analysis. The XRPD spectrum is shown in FIG8 , and the positions of the characteristic peaks are shown in Table 11.

[0208] The DSC spectrum showed that the endothermic peak was 228.04℃.

[0209] The TGA spectrum showed that the weight loss was 0.01% at 30℃-150℃.

[0210] DVS testing showed that under normal storage conditions (i.e., 25°C, 60% RH), the sample experienced a moisture gain of approximately 0.04%. Under accelerated storage conditions (i.e., 70% RH), the moisture gain was approximately 0.06%. Under extreme conditions (i.e., 90% RH), the moisture gain was approximately 0.15%. Furthermore, subsequent DVS testing revealed no change in the crystal form.

[0211] Table 11

[0212] Example 17: Preparation of Hydrobromide Form I

[0213] About 10 mg of compound 1 was weighed and dissolved in 0.15 mL of acetonitrile. Hydrobromic acid (2 mol / L, 16.4 μL) was added and stirred for crystallization. After centrifugation, the solid was vacuum dried to obtain the title product.

[0214] Example 18: Preparation of Hydrobromide Crystal Form I

[0215] About 10 mg of compound 1 was weighed and dissolved in 0.15 mL of acetone. Hydrobromic acid (2 mol / L, 16.4 μL) was added and stirred for crystallization. After centrifugation, the solid was vacuum dried to obtain the title product.

[0216] Example 19: Preparation of Hydrochloride Form I

[0217] About 10 mg of compound 1 was weighed and dissolved in 0.15 mL of ethanol. Hydrochloric acid (2 mol / L, 16.4 μL) was added and stirred for crystallization. After centrifugation, the solid was vacuum dried to obtain the product.

[0218] The product was defined as hydrochloride form I by X-ray powder diffraction analysis. The XRPD spectrum is shown in FIG9 , and the positions of the characteristic peaks are shown in Table 12.

[0219] The DSC spectrum showed that the endothermic peak was 173.40℃.

[0220] The TGA spectrum showed that the weight loss was 4.16% at 30℃-125℃.

[0221] DVS testing showed that under normal storage conditions (i.e., 25°C, 60% RH), the sample experienced a moisture gain of approximately 0.33%. Under accelerated storage conditions (i.e., 70% RH), the moisture gain was approximately 0.37%. Under extreme conditions (i.e., 90% RH), the moisture gain was approximately 0.53%. Furthermore, subsequent DVS testing revealed no change in the crystal form.

[0222] Table 12

[0223] Example 20: Preparation of Hydrochloride Form I

[0224] About 200 mg of compound 1 was weighed and dissolved in 0.8 mL of acetone. Concentrated hydrochloric acid (12 mol / L, 53 μL) was added and stirred for crystallization. After centrifugation, the solid was vacuum dried to obtain the title product.

[0225] Example 21: Preparation of Hydrochloride Form I

[0226] About 40 mg of compound 1 was weighed and dissolved in 0.2 mL of acetone. A hydrochloric acid acetone solution (2 mol / L, 62.8 μL) was added, and the mixture was stirred for crystallization. After centrifugation, the solid was vacuum dried to obtain the title product.

[0227] Example 22: Preparation of Hydrochloride Form I

[0228] About 10 mg of compound 1 was weighed and dissolved in 0.15 mL of acetonitrile. Hydrochloric acid (2 mol / L, 16.4 μL) was added and stirred for crystallization. After centrifugation, the solid was vacuum dried to obtain the product.

[0229] Example 23: Preparation of Hydrochloride II Crystalline Form

[0230] About 10 mg of compound 1 was weighed, added to 0.15 mL of 10% water / methanol, and hydrochloric acid (2 mol / L, 16.4 μL) was added, stirred for crystallization, centrifuged, and the solid was vacuum dried to obtain the product.

[0231] The product was defined as hydrochloride II crystal form by X-ray powder diffraction analysis. The XRPD spectrum is shown in FIG10 , and the positions of its characteristic peaks are shown in Table 13.

[0232] The DSC spectrum showed that the endothermic peak was 172.83℃.

[0233] The TGA spectrum showed that the weight loss was 0.40% at 30℃-120℃.

[0234] DVS testing showed that under normal storage conditions (i.e., 25°C, 60% RH), the sample experienced a moisture gain of approximately 0.08%. Under accelerated storage conditions (i.e., 70% RH), the moisture gain was approximately 0.12%. Under extreme conditions (i.e., 90% RH), the moisture gain was approximately 0.23%. Furthermore, subsequent DVS testing revealed no change in the crystal form.

[0235] Table 13

[0236] Test Example 2: Influencing Factor Experiment

[0237] The hydrobromide salt form I, p-toluenesulfonate salt form I, hydrochloride salt form I, hydrochloride salt form II, maleate salt form II, phosphate salt form I, and sulfate salt form II were laid out in the open air, and the stability of the samples was investigated under light (4500 Lux), high temperature (40°C, 60°C), and high humidity (RH75%, RH92.5%) conditions, respectively. The sampling period was 30 days.

[0238] Table 14: Factors affecting stability of hydrobromide salt I crystal form

[0239] Table 15: Factors affecting stability of p-toluenesulfonate I crystalline form

[0240] Table 16: Factors affecting stability of hydrochloride salt I crystal form

[0241] Table 17: Factors affecting stability of hydrochloride II crystal form

[0242] Table 18: Factors affecting the stability of maleate II crystal form

[0243] Table 19: Factors affecting the stability of phosphate I crystal form

[0244] Table 20: Factors affecting the stability of sulfate II crystal form

[0245] Conclusion: The influencing factor experiment showed that the phosphate I crystal form, sulfate II crystal form, maleate II crystal form, hydrochloride II crystal form, p-toluenesulfonate I crystal form and hydrobromide I crystal form had good physical and chemical stability.

[0246] Test Example 3: Long-term accelerated stability investigation

[0247] The hydrobromide salt form I, p-toluenesulfonate salt form I, hydrochloride salt form I, hydrochloride salt form II, maleate salt form II, phosphate salt form I and sulfate salt form II were placed under 25°C / 60% RH and 40°C / 75% RH conditions, respectively, to investigate their stability.

[0248] Table 21: Long-term accelerated stability of hydrobromide salt form I

[0249] Table 22: Long-term accelerated stability of p-toluenesulfonate Form I

[0250] Table 23: Long-term accelerated stability of hydrochloride salt form I

[0251] Table 24: Long-term accelerated stability of hydrochloride salt II crystal form

[0252] Table 25: Long-term accelerated stability of maleate II crystal form

[0253] Table 26: Long-term accelerated stability of phosphate form I

[0254] Table 27: Long-term accelerated stability of sulfate II crystal form

Claims

1. A pharmaceutically acceptable salt of the compound 8-chloro-N-(2,2-difluorobenzo[d][1,3]dioxolan-5-yl)quinolin-2-amine, characterized in that: The pharmaceutically acceptable salt is selected from the group consisting of p-toluenesulfonate, phosphate, sulfate, maleate, hydrobromide and hydrochloride.

2. The pharmaceutically acceptable salt according to claim 1, characterized in that The chemical ratio of the compound 8-chloro-N-(2,2-difluorobenzo[d][1,3]dioxolan-5-yl)quinolin-2-amine to the acid molecule is 1:0.5 to 1:3, preferably 1:0.5, 1:1, 1:2 or 1:3, and most preferably 1:0.5 or 1:

1.

3. A method for preparing the pharmaceutically acceptable salt according to claim 1 or 2, characterized in that: The method comprises the steps of forming a salt of a compound 8-chloro-N-(2,2-difluorobenzo[d][1,3]dioxolan-5-yl)quinolin-2-amine with an acid.

4. The method according to claim 3, characterized in that The solvent used in the salt-forming reaction is selected from at least one of ethanol, acetonitrile, acetone, methanol and water.

5. The p-toluenesulfonate salt of compound 8-chloro-N-(2,2-difluorobenzo[d][1,3]dioxolan-5-yl)quinolin-2-amine in crystalline form I, characterized in that: The X-ray powder diffraction pattern, expressed as a diffraction angle of 2θ, has characteristic peaks at 6.616, 8.996, 14.120, 19.138 and 22.592, preferably has characteristic peaks at 6.616, 8.996, 14.120, 17.120, 19.138, 20.615, 22.592 and 25.033, more preferably has characteristic peaks at 6.616, 8.996, 13.304, 14.120, 17.120, 19.138, 20.615, 21.289, 22.592, 25.033, 26.101 and 26.503, and most preferably has an X-ray powder diffraction pattern as shown in Figure 1, expressed as a diffraction angle of 2θ.

6. A crystalline form I of the phosphate salt of the compound 8-chloro-N-(2,2-difluorobenzo[d][1,3]dioxolan-5-yl)quinolin-2-amine, characterized in that: The X-ray powder diffraction pattern, expressed as a diffraction angle of 2θ, has characteristic peaks at 6.511, 9.685, 12.660, 14.012 and 15.704, preferably has characteristic peaks at 6.511, 9.685, 12.660, 14.012, 15.704, 16.517, 21.487 and 22.209, more preferably has characteristic peaks at 6.511, 9.685, 12.660, 14.012, 15.704, 16.517, 18.507, 21.487, 22.209, 24.493 and 25.596, and most preferably the X-ray powder diffraction pattern, expressed as a diffraction angle of 2θ, is shown in Figure 2.

7. The sulfate salt of compound 8-chloro-N-(2,2-difluorobenzo[d][1,3]dioxolan-5-yl)quinolin-2-amine, crystalline form I, characterized in that: The X-ray powder diffraction pattern, expressed as a diffraction angle of 2θ, has characteristic peaks at 5.687, 11.448, 14.670, 17.315, 23.323 and 24.907, preferably has characteristic peaks at 5.687, 9.540, 11.448, 14.670, 17.315, 18.906, 23.323 and 24.907, more preferably has characteristic peaks at 5.687, 9.540, 11.448, 12.137, 14.670, 17.315, 17.465, 18.906, 21.682, 23.323 and 24.907, and most preferably has an X-ray powder diffraction pattern as shown in Figure 3, expressed as a diffraction angle of 2θ.

8. A crystalline form II of the sulfate salt of the compound 8-chloro-N-(2,2-difluorobenzo[d][1,3]dioxolan-5-yl)quinolin-2-amine, characterized in that: The X-ray powder diffraction pattern expressed as a diffraction angle 2θ has characteristic peaks at 11.895, 13.102, 15.607, 16.658 and 17.967, preferably has characteristic peaks at 6.518, 11.895, 13.102, 15.607, 16.181, 16.658, 17.967 and 24.329, more preferably has characteristic peaks at 6.518, 10.808, 11.291, 11.895, 13.102, 15.196, 15.607, 16.181, 16.658, 17.967 and 24.329, and most preferably the X-ray powder diffraction pattern expressed as a diffraction angle 2θ is shown in Figure 4.

9. A sulfate salt of compound 8-chloro-N-(2,2-difluorobenzo[d][1,3]dioxolan-5-yl)quinolin-2-amine in form III, characterized in that: The X-ray powder diffraction pattern expressed in terms of a diffraction angle of 2θ has characteristic peaks at 13.502, 16.610, 17.858, 22.247 and 26.671, preferably has characteristic peaks at 8.675, 13.502, 16.610, 17.858, 22.247, 25.379, 26.671 and 27.029, more preferably has characteristic peaks at 8.675, 13.502, 16.610, 17.449, 17.858, 22.247, 24.292, 25.379, 26.671 and 27.029, and most preferably has an X-ray powder diffraction pattern expressed in terms of a diffraction angle of 2θ as shown in Figure 5.

10. A maleate salt of compound 8-chloro-N-(2,2-difluorobenzo[d][1,3]dioxolan-5-yl)quinolin-2-amine in form I, characterized in that: The X-ray powder diffraction pattern expressed in terms of a diffraction angle of 2θ has characteristic peaks at 8.101, 12.810, 16.435, 26.186 and 27.616, preferably at 8.101, 12.810, 16.435, 20.667, 25.309, 26.186 and 27.616, more preferably at 8.101, 12.810, 14.598, 16.435, 20.667, 21.474, 22.460, 25.309, 26.186 and 27.616, and most preferably the X-ray powder diffraction pattern expressed in terms of a diffraction angle of 2θ is shown in FIG6 .

11. A maleate II crystalline form of the compound 8-chloro-N-(2,2-difluorobenzo[d][1,3]dioxolan-5-yl)quinolin-2-amine, characterized in that: The X-ray powder diffraction pattern expressed as a diffraction angle 2θ has characteristic peaks at 7.817, 8.777, 11.663, 16.222 and 26.051, preferably at 7.817, 8.777, 11.663, 12.691, 16.222, 21.791 and 26.051, more preferably at 7.817, 8.451, 8.777, 11.663, 12.691, 15.028, 16.222, 17.399, 21.791 and 26.051, and most preferably the X-ray powder diffraction pattern expressed as a diffraction angle 2θ is shown in Figure 7.

12. A crystalline form I of the hydrobromide salt of the compound 8-chloro-N-(2,2-difluorobenzo[d][1,3]dioxolan-5-yl)quinolin-2-amine, characterized in that: The X-ray powder diffraction pattern expressed in terms of a diffraction angle of 2θ has characteristic peaks at 6.077, 12.258, 22.769, 24.662 and 26.579, preferably has characteristic peaks at 6.077, 12.258, 22.769, 24.662, 25.696, 26.579 and 34.105, more preferably has characteristic peaks at 6.077, 12.258, 16.589, 20.396, 22.769, 24.662, 25.218, 25.696, 26.579 and 34.105, and most preferably the X-ray powder diffraction pattern expressed in terms of a diffraction angle of 2θ is shown in FIG8 .

13. The hydrochloride salt of compound 8-chloro-N-(2,2-difluorobenzo[d][1,3]dioxolan-5-yl)quinolin-2-amine, in form I, characterized in that: The X-ray powder diffraction pattern expressed as a diffraction angle 2θ has characteristic peaks at 13.229, 14.520, 18.561, 19.925 and 23.830, preferably at 13.229, 14.520, 18.561, 19.925, 23.830, 24.603 and 25.325, more preferably at 13.229, 14.520, 18.561, 19.925, 21.085, 22.588, 23.830, 24.603, 25.325 and 26.708, and most preferably the X-ray powder diffraction pattern expressed as a diffraction angle 2θ is shown in Figure 9.

14. The hydrochloride salt of compound 8-chloro-N-(2,2-difluorobenzo[d][1,3]dioxolan-5-yl)quinolin-2-amine, in form II, characterized in that: The X-ray powder diffraction pattern expressed in terms of a diffraction angle of 2θ has characteristic peaks at 11.888, 13.593, 17.851, 23.977 and 26.309, preferably at 11.888, 13.593, 17.851, 19.114, 23.977, 25.878 and 26.309, more preferably at 11.888, 13.593, 14.492, 17.851, 19.114, 20.367, 23.977, 25.878, 26.309 and 29.779, and most preferably the X-ray powder diffraction pattern expressed in terms of a diffraction angle of 2θ is shown in Figure 10.

15. The crystal form according to any one of claims 5 to 14, characterized in that The error range of the 2θ value is ±0.

2.

16. A pharmaceutical composition comprising the pharmaceutically acceptable salt according to claim 1 or 2, or the crystal form according to any one of claims 5 to 15, and a pharmaceutically acceptable excipient.

17. A pharmaceutical composition prepared from the pharmaceutically acceptable salt according to claim 1 or 2, or the crystal form according to any one of claims 5 to 15, and a pharmaceutically acceptable excipient.

18. Use of the pharmaceutically acceptable salt of claim 1 or 2, or the crystalline form of any one of claims 5 to 15, or the pharmaceutical composition of claim 16 or 17 in the preparation of a medicament for treating and / or preventing a disease or condition, wherein the disease or condition is selected from inflammation and cancer, the inflammation is preferably inflammatory bowel disease, and the cancer is preferably melanoma or breast cancer.

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