Quinoline amine compound crystal form and preparation method thereof

Abstract

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

Description

A quinolineamine compound crystal form and preparation method thereof

[0001] This application claims the benefit of Chinese Patent Application No. 2022114917970, filed on November 25, 2022. This application incorporates the entirety of the aforementioned Chinese Patent Application. Technical Field

[0002] The present invention relates to a quinolineamine compound crystal form and a preparation method thereof. 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] The crystal structure of a pharmaceutically active ingredient often affects the chemical and physical stability of the drug. Differences in crystallization and storage conditions can lead to variations in the compound's crystal structure, sometimes resulting in the formation of alternative crystalline forms. Generally speaking, amorphous pharmaceutical products lack a regular crystal structure and often exhibit other drawbacks, such as poor product stability, difficulty in filtration, susceptibility to agglomeration, and poor flowability. Therefore, studying their crystal forms is crucial for developing drugs suitable for industrial production and possessing excellent biological activity.

[0007] Summary of the Invention

[0008] On the one hand, the present disclosure provides a crystalline form A 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 2θ, with characteristic peaks at 7.237, 9.232, 13.702, 14.459 and 18.917.

[0009] In some embodiments, the Form A has an X-ray powder diffraction pattern expressed as a diffraction angle 2θ, with characteristic peaks at 7.237, 9.232, 13.702, 14.459, 18.917, 24.428 and 29.321.

[0010] In some embodiments, the Form A has an X-ray powder diffraction pattern expressed as a diffraction angle 2θ, with characteristic peaks at 7.237, 9.232, 13.702, 14.459, 18.033, 18.917, 24.428, 25.521 and 29.321.

[0011] In other embodiments, the X-ray powder diffraction pattern of the crystal form A expressed in terms of a diffraction angle 2θ is shown in FIG1 .

[0012] The present disclosure also provides a method for preparing the aforementioned crystalline form of Compound A, which is selected from any one of the following methods:

[0013] Method 1:

[0014] (a) mixing the compound 8-chloro-N-(2,2-difluorobenzo[d][1,3]dioxolan-5-yl)quinolin-2-amine with the solvent (1), stirring and dissolving the mixture or heating the mixture to dissolve the mixture,

[0015] (b) adding solvent (2) and crystallizing,

[0016] wherein solvent (1) is selected from acetonitrile, methanol, ethanol, isopropanol, acetone, ethyl acetate, isopropyl acetate, tetrahydrofuran, methyl isopropyl ketone, dichloromethane, 10% water / methanol, 7% water / ethanol, 10% water / isopropanol or 10% water / acetone, and solvent (2) is selected from water, cyclohexane or n-heptane;

[0017] Or, method 2:

[0018] (a) mixing the compound 8-chloro-N-(2,2-difluorobenzo[d][1,3]dioxolan-5-yl)quinolin-2-amine with the solvent (3), stirring and dissolving the mixture or heating the mixture to dissolve the mixture,

[0019] (b) crystallization,

[0020] wherein the solvent (3) is selected from tetrahydrofuran, ethyl acetate or dichloromethane;

[0021] Or, method three:

[0022] (a) mixing the compound 8-chloro-N-(2,2-difluorobenzo[d][1,3]dioxolan-5-yl)quinolin-2-amine with a solvent (4),

[0023] (b) stirring and slurrying, wherein the solvent (4) is selected from water, cyclohexane, n-heptane, methanol, ethanol, isopropanol, dichloromethane, 1,4-dioxane, 10% water / methanol, 7% water / ethanol or 10% water / isopropanol.

[0024] On the one hand, the present disclosure provides a crystalline form B 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 2θ, with characteristic peaks at 8.205, 9.781, 12.87, 15.907 and 19.796.

[0025] In some embodiments, the B crystalline form has an X-ray powder diffraction pattern expressed as a diffraction angle 2θ, with characteristic peaks at 8.205, 9.781, 12.870, 15.907, 19.448, 19.796, 20.264 and 23.185.

[0026] In some embodiments, the B crystalline form has an X-ray powder diffraction pattern expressed as a diffraction angle 2θ, with characteristic peaks at 8.205, 9.781, 10.672, 12.87, 15.356, 15.907, 16.997, 19.448, 19.796, 20.264 and 23.185.

[0027] In other embodiments, the X-ray powder diffraction pattern of the B crystal form expressed in terms of a diffraction angle 2θ is shown in FIG2 .

[0028] The present disclosure also provides a method for preparing the aforementioned crystalline form of Compound B, the method comprising:

[0029] (a) mixing the compound 8-chloro-N-(2,2-difluorobenzo[d][1,3]dioxolan-5-yl)quinolin-2-amine with a solvent (5), stirring and dissolving the mixture or heating the mixture to dissolve the mixture;

[0030] (b) adding solvent (6) and crystallizing,

[0031] The solvent (5) is selected from dimethyl sulfoxide, and the solvent (6) is selected from water.

[0032] On the one hand, the present disclosure provides a crystalline form C 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 2θ, with characteristic peaks at 9.296, 15.522, 18.784, 23.216 and 25.889.

[0033] In some embodiments, the Form C has an X-ray powder diffraction pattern expressed as a diffraction angle 2θ, with characteristic peaks at 7.753, 9.296, 15.522, 18.784, 21.398, 23.216 and 25.889.

[0034] In some embodiments, the Form C has an X-ray powder diffraction pattern expressed as a diffraction angle 2θ, with characteristic peaks at 7.353, 7.753, 9.296, 14.475, 15.522, 16.248, 17.320, 18.784, 21.398, 23.216 and 25.889.

[0035] In other embodiments, the X-ray powder diffraction pattern of the Form C expressed in terms of a diffraction angle of 2θ is shown in FIG3 .

[0036] The present disclosure also provides a method for preparing the aforementioned crystalline form of Compound C, the method comprising:

[0037] (a) mixing the compound 8-chloro-N-(2,2-difluorobenzo[d][1,3]dioxolan-5-yl)quinolin-2-amine with a solvent (7), stirring and dissolving the mixture or heating the mixture to dissolve the mixture;

[0038] (b) adding solvent (8) and crystallizing,

[0039] The solvent (7) is selected from 1,4-dioxane, and the solvent (8) is selected from n-heptane.

[0040] On the one hand, the present disclosure provides a D-type 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 7.350, 12.084, 15.384, 18.643, and 29.312.

[0041] In some embodiments, the D crystalline form has an X-ray powder diffraction pattern expressed as a diffraction angle 2θ, with characteristic peaks at 7.350, 12.084, 15.384, 16.260, 17.855, 18.643, 21.610 and 29.312.

[0042] In some embodiments, the D crystalline form has an X-ray powder diffraction pattern expressed as a diffraction angle 2θ, with characteristic peaks at 7.350, 12.084, 15.384, 16.260, 17.855, 18.643, 21.610, 22.768, 24.347, 25.201, 26.038 and 29.312.

[0043] In other embodiments, the X-ray powder diffraction pattern of the D crystal form expressed in terms of a diffraction angle of 2θ is shown in FIG4 .

[0044] The present disclosure also provides a method for preparing the aforementioned crystalline form of compound D, the method comprising:

[0045] (a) mixing the compound 8-chloro-N-(2,2-difluorobenzo[d][1,3]dioxolan-5-yl)quinolin-2-amine with a solvent (9), stirring and dissolving the mixture or heating the mixture to dissolve the mixture;

[0046] (b) adding solvent (10) and crystallizing,

[0047] The solvent (9) is selected from tetrahydrofuran, and the solvent (10) is selected from water.

[0048] On the one hand, the present disclosure provides the E 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 13.489, 18.000, 23.559, 24.276 and 26.328.

[0049] In some embodiments, the E crystalline form has an X-ray powder diffraction pattern expressed as a diffraction angle 2θ, with characteristic peaks at 13.489, 16.863, 18.000, 23.559, 24.276, 26.108, 26.328 and 27.094.

[0050] In some embodiments, the E crystalline form has an X-ray powder diffraction pattern expressed as a diffraction angle 2θ, with characteristic peaks at 13.489, 14.587, 16.863, 18.000, 18.943, 23.279, 23.559, 24.276, 25.768, 26.108, 26.328 and 27.094.

[0051] In other embodiments, the X-ray powder diffraction pattern of the E crystal form expressed in terms of a diffraction angle of 2θ is shown in FIG5 .

[0052] On the one hand, the present disclosure provides a crystalline form F 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 2θ, with characteristic peaks at 9.299, 14.439, 15.621, 16.200 and 17.314.

[0053] In some embodiments, the F crystalline form has an X-ray powder diffraction pattern expressed as a diffraction angle 2θ, with characteristic peaks at 7.734, 9.299, 14.439, 15.621, 16.200, 17.314, 21.395 and 25.814.

[0054] In some embodiments, the F crystalline form has an X-ray powder diffraction pattern expressed as a diffraction angle 2θ, with characteristic peaks at 7.734, 9.299, 14.439, 15.621, 16.200, 17.314, 21.055, 21.395, 23.194, 25.814, 28.906, 34.485 and 43.544.

[0055] In other embodiments, the X-ray powder diffraction pattern of the F crystal form expressed in terms of a diffraction angle 2θ is shown in FIG6 .

[0056] Furthermore, the X-ray powder diffraction pattern of the compound in the present disclosure, crystalline form A, crystalline form B, crystalline form C, crystalline form D, crystalline form E or crystalline form F, is expressed in terms of a diffraction angle 2θ, wherein the error range of the 2θ angle is ±0.2.

[0057] In certain embodiments, the method for preparing the crystalline form described in the present disclosure further comprises any one of the steps of filtering, washing or drying.

[0058] In some embodiments, the crystallization includes but is not limited to stirring crystallization (dissolution crystallization, slurry crystallization) and volatile crystallization.

[0059] 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.

[0060] On the other hand, the present disclosure also provides a pharmaceutical composition comprising the aforementioned crystal form and a pharmaceutically acceptable excipient.

[0061] The present disclosure also provides a pharmaceutical composition prepared from the aforementioned crystal form and a pharmaceutically acceptable excipient.

[0062] 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.

[0063] 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.

[0064] 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.

[0065] In some embodiments, the inflammation is inflammatory bowel disease. In some embodiments, the cancer is melanoma or breast cancer.

[0066] 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.

[0067] 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.

[0068] 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.

[0069] 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.

[0070] 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.

[0071] The crystalline forms disclosed herein include, but are not limited to, solvates of the compound represented by formula (I), wherein the solvents include, but are not limited to, water. BRIEF DESCRIPTION OF THE DRAWINGS

[0072] Figure 1 is the XRPD spectrum of Form A of Compound 1.

[0073] Figure 2 is the XRPD spectrum of Form B of Compound 1.

[0074] Figure 3 is the XRPD spectrum of Form C of Compound 1.

[0075] Figure 4 is the XRPD spectrum of Form D of Compound 1.

[0076] Figure 5 is the XRPD spectrum of Form E of Compound 1.

[0077] Figure 6 is the XRPD spectrum of Form F of Compound 1. DETAILED DESCRIPTION

[0078] 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.

[0079] Test conditions of the instruments used in the experiment:

[0080] 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.

[0081] 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).

[0082] 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).

[0083] 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°.

[0084] 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 or from 25 to 350°C, and a nitrogen purge rate of 50 mL / min.

[0085] 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.

[0086] 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.

[0087] 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.

[0088] 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.

[0089] 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)

[0090] 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) 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).

[0091] MS m / z(ESI):335.0[M+1].

[0092] 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).

[0093] Test Example 1. Upregulation of miR-124

[0094] 1. Experimental Materials and Instruments

[0095] 1. Dynabead Human T-Activator CD3 / CD28 for T Cell Expansion and Activation (Gibco, 11131D)

[0096] 2. Pan T Cell Isolation Kit, human (Miltenyi, 130-096-535)

[0097] 3. Human interleukin-2 (Human IL-2) (Peprotech, 200-02-100)

[0098] 4. Small RNA extraction kit (microRNA extraction kit) (Qiagen, 217004)

[0099] 5. miScript II RT Kit (Qiagen, 218161)

[0100] 6. miScript SYBR Green PCR Kit (Qiagen, 218073)

[0101] 7. Phosphate buffer PBS, pH 7.4 (Shanghai Yuanpei Biotechnology Co., Ltd., B320)

[0102] 8. Bovine serum albumin, BSA (Biyuntian, ST023)

[0103] 9.EDTA (0.5M), pH 8.0 (Invitrogen, AM9260G)

[0104] 10. LS Columns (Miltenyi, 130-042-401)

[0105] 11. 24-well cell culture plate (Corning, 3524)

[0106] 12. 96-well plate (Corning, 3788)

[0107] 13. Cell culture incubator (Thermo, Stericycle i160)

[0108] 14. Real-time fluorescence quantitative PCR instrument (Applied biosystem, QuantStudio6Flex)

[0109] 15.PCR instrument (Applied biosystem, ProFlex)

[0110] 16. 96-well clear PCR plate, 0.2 mL (Applied biosystems, N8010560)

[0111] 17. RPMI1640 culture medium (Gibco, 11875119)

[0112] 18. Fetal bovine serum, FBS (Gibco, 10099-141)

[0113] 19. Magnetic rack (Invitrogen, DynaMag TM -2)

[0114] 20. Six-well cell culture plate (Thermo, 150239)

[0115] 21. Spectrophotometer (IMPLEN, NP80)

[0116] 22. Magnetic bead separation rack (QuadroMACS Separator) (Miltenyi Biotec, 130-090-976)

[0117] 23.miR124-3P-F primer (customized by Genewise)

[0118] 24.hsa-U6 detection primer (Tian Gen, CD201-0145)

[0119] 2. Experimental steps

[0120] 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.

[0121] 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.

[0122] 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.

[0123] 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.

[0124] 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.

[0125] 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.

[0126] 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.

[0127] Table 1 Fluorescence quantitative PCR reaction components

[0128] Table 2 Fluorescence quantitative PCR steps

[0129] Table 3 Fluorescence quantitative PCR detection primer list

[0130] 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)]).

[0131] Compound 1 upregulated miR-124 by 3.9 times (fold), and had good activity in promoting miR124 upregulation.

[0132] Example 2: Preparation of Crystal Form A

[0133] 250 mg of compound 1 was added to 2.5 mL of acetonitrile and stirred to dissolve. 17.5 mL of water was added and stirred to crystallize. The mixture was slurried at room temperature for 4 days, filtered, and vacuum dried to obtain a solid.

[0134] The product was defined as Form A by X-ray powder diffraction analysis. The X-ray powder diffraction data are shown in Table 4, and the X-ray powder diffraction spectrum is shown in FIG1.

[0135] The DSC spectrum showed an endothermic peak at 177.14°C.

[0136] The TGA spectrum showed a weight loss of 0.22% from 30°C to 100°C.

[0137] DVS testing showed that under normal storage conditions (i.e., room temperature, 60% RH), the sample experienced a moisture absorption weight gain of approximately 0.02%. Under accelerated storage conditions (i.e., 70% RH), the moisture absorption weight gain was approximately 0.02%. Under extreme conditions (i.e., 90% RH), the moisture absorption weight gain was approximately 0.18%. Furthermore, subsequent DVS testing revealed no change in the crystal form.

[0138] Table 4

[0139] Example 3: Preparation of Crystal Form A

[0140] Compound 1 (6 mg, 17.63 μmol) was dissolved in 0.3 mL of tetrahydrofuran, and after the solution became clear, the solid was slowly evaporated to obtain a solid, which was then dried in vacuo at 45° C. for 3 hours to obtain the product.

[0141] X-ray powder diffraction analysis showed that the product was crystal form A.

[0142] Example 4: Preparation of Crystal Form A

[0143] Compound 1 (11 mg, 32.87 μmol) was dissolved in 0.4 mL of ethyl acetate, and the solid was evaporated slowly after dissolving clearly. The solid was dried in vacuo at 45° C. for 3 hours to obtain the product.

[0144] X-ray powder diffraction analysis showed that the product was crystal form A.

[0145] Example 5: Preparation of Crystal Form A

[0146] Compound 1 (14 mg, 41.83 μmol) was dissolved in 0.5 mL of dichloromethane, and the solid was evaporated slowly after dissolving. The solid was dried in vacuo at 45° C. for 3 hours to obtain the product.

[0147] X-ray powder diffraction analysis showed that the product was crystal form A.

[0148] Example 6: Preparation of Crystal Form A

[0149] Compound 1 (1.70 g, 5.08 mmol) was dispersed in 35 mL of cyclohexane, stirred and slurried for 48 hours, filtered and the filter cake was collected, and vacuum dried at 45° C. for 16 hours to obtain the product.

[0150] X-ray powder diffraction analysis showed that the product was crystal form A.

[0151] Example 7: Preparation of Crystal Form A

[0152] 10 mg of compound 1 was weighed, 1 ml of purified water was added and slurried at room temperature for 3 days, and the solid was filtered and vacuum dried to obtain the product.

[0153] X-ray powder diffraction analysis showed that the product was crystal form A.

[0154] Example 8: Preparation of Crystal Form A

[0155] According to the method of Example 7, 10 mg of compound 1 was weighed, added to a solvent, slurried, filtered, and the solid was vacuum dried to obtain the product.

[0156] X-ray powder diffraction analysis showed that the product was crystal form A. The data are shown in Table 5.

[0157] Table 5

[0158] Example 9: Preparation of Crystal Form A

[0159] 10 mg of compound 1 was weighed, 0.2 ml of methanol was added and stirred at room temperature to dissolve, 0.2 ml of purified water was added and stirred to crystallize, and the mixture was slurried at room temperature for 1 day. After filtering, the solid was vacuum dried to obtain the product.

[0160] X-ray powder diffraction analysis showed that the product was crystal form A.

[0161] Example 10: Preparation of Crystal Form A

[0162] 10 mg of compound 1 was weighed, 0.2 ml of ethanol was added and stirred at room temperature to dissolve, 0.4 ml of n-heptane was added and stirred for 1 day, and the product was evaporated and crystallized to obtain the product.

[0163] X-ray powder diffraction analysis showed that the product was crystal form A.

[0164] Example 11: Preparation of Crystal Form A

[0165] 10 mg of compound 1 was weighed, 0.6 ml of isopropanol was added and stirred at room temperature to dissolve, 0.4 ml of n-heptane was added and stirred for 1 day, and the product was evaporated and crystallized to obtain the product.

[0166] X-ray powder diffraction analysis showed that the product was crystal form A.

[0167] Example 12: Preparation of Crystal Form A

[0168] 10 mg of compound 1 was weighed, 0.1 ml of acetone was added and stirred at room temperature to dissolve, 0.4 ml of n-heptane was added and stirred for 1 day, and the product was evaporated and crystallized to obtain the product.

[0169] X-ray powder diffraction analysis showed that the product was crystal form A.

[0170] Example 13: Preparation of Crystal Form A

[0171] 10 mg of compound 1 was weighed, 0.1 ml of ethyl acetate was added and stirred at room temperature to dissolve, 0.4 ml of n-heptane was added and stirred for 1 day, and the product was evaporated and crystallized to obtain the product.

[0172] X-ray powder diffraction analysis showed that the product was crystal form A.

[0173] Example 14: Preparation of Crystal Form A

[0174] Weigh 10 mg of compound 1, add 0.1 ml of isopropyl acetate and stir at room temperature to dissolve, add 0.4 ml of n-heptane and stir to crystallize, beat at room temperature for 1 day, add 0.2 ml of n-heptane and continue beating for 3 hours, filter and vacuum dry the solid to obtain the product.

[0175] X-ray powder diffraction analysis showed that the product was crystal form A.

[0176] Example 15: Preparation of Crystal Form A

[0177] 10 mg of compound 1 was weighed, 0.1 ml of tetrahydrofuran was added and stirred at room temperature to dissolve, 0.4 ml of n-heptane was added and stirred for 1 day, and the product was evaporated and crystallized to obtain the product.

[0178] X-ray powder diffraction analysis showed that the product was crystal form A.

[0179] Example 16: Preparation of Crystal Form A

[0180] 10 mg of compound 1 was weighed, 0.1 ml of methyl isopropyl ketone was added, and the mixture was stirred at room temperature to dissolve, 0.4 ml of n-heptane was added, and the mixture was stirred for 1 day. The product was evaporated and crystallized to obtain the product.

[0181] X-ray powder diffraction analysis showed that the product was crystal form A.

[0182] Example 17: Preparation of Crystal Form A

[0183] 10 mg of compound 1 was weighed, 0.4 ml of dichloromethane was added and stirred at room temperature to dissolve, 0.4 ml of n-heptane was added for crystallization, and the mixture was slurried at room temperature for 1 day. 0.2 ml of n-heptane was added and the mixture was slurried for 3 hours. After filtration, the solid was vacuum dried to obtain the product.

[0184] X-ray powder diffraction analysis showed that the product was crystal form A.

[0185] Example 18: Preparation of Crystal Form A

[0186] 10 mg of compound 1 was weighed, 1 ml of 10% water / methanol was added and stirred at room temperature to dissolve, 0.1 ml of water was added and stirred to crystallize, 0.1 ml of water was added and slurrying was continued for 1 day, and the solid was filtered and vacuum dried to obtain the product.

[0187] X-ray powder diffraction analysis showed that the product was crystal form A.

[0188] Example 19: Preparation of Crystal Form A

[0189] 10 mg of compound 1 was weighed, added to 0.6 ml of 7% water / ethanol, stirred at room temperature to dissolve, and 0.4 ml of n-heptane was added, evaporated and crystallized to obtain the product.

[0190] X-ray powder diffraction analysis showed that the product was crystal form A.

[0191] Example 20: Preparation of Crystal Form A

[0192] 10 mg of compound 1 was weighed, 0.6 ml of 10% water / isopropanol was added, and the mixture was stirred at room temperature to dissolve, 0.4 ml of n-heptane was added, and the product was evaporated and crystallized to obtain the product.

[0193] X-ray powder diffraction analysis showed that the product was crystal form A.

[0194] Example 21: Preparation of Crystal Form A

[0195] 10 mg of compound 1 was weighed, 0.1 ml of 10% water / acetone was added, and the mixture was stirred at room temperature to dissolve, 0.4 ml of n-heptane was added, and the product was evaporated and crystallized to obtain the product.

[0196] X-ray powder diffraction analysis showed that the product was crystal form A.

[0197] Example 22: Preparation of Crystal Form A

[0198] 10 mg of compound 1 was weighed, 0.2 ml of ethanol was added and stirred at room temperature to dissolve, 0.1 ml of purified water was added and stirred to precipitate, 0.2 ml of purified water was added and stirred for 1 day, and the solid was filtered and vacuum dried to obtain the product.

[0199] X-ray powder diffraction analysis showed that the product was crystal form A.

[0200] Example 23: Preparation of Crystal Form A

[0201] 10 mg of compound 1 was weighed, 0.2 ml of ethanol was added and stirred at room temperature to dissolve, 0.4 ml of cyclohexane was added and stirred for 1 day, and then evaporated and crystallized to obtain the product.

[0202] X-ray powder diffraction analysis showed that the product was crystal form A.

[0203] Example 24: Preparation of Crystal Form A

[0204] 10 mg of compound 1 was weighed, 0.1 ml of acetone was added and stirred at room temperature to dissolve, 0.1 ml of purified water was added and stirred to precipitate, 0.2 ml of purified water was added and stirred for 1 day, and the solid was filtered and vacuum dried to obtain the product.

[0205] X-ray powder diffraction analysis showed that the product was crystal form A.

[0206] Example 25: Preparation of Crystal Form A

[0207] 10 mg of compound 1 was weighed, 0.1 ml of acetone was added and stirred at room temperature to dissolve, 0.4 ml of cyclohexane was added and stirred for 1 day, and then evaporated and crystallized to obtain the product.

[0208] X-ray powder diffraction analysis showed that the product was crystal form A.

[0209] Example 26: Preparation of Crystal Form A

[0210] 10 mg of compound 1 was weighed, 0.1 ml of ethyl acetate was added and stirred at room temperature to dissolve, 0.4 ml of cyclohexane was added and stirred for 1 day, and then evaporated and crystallized to obtain the product.

[0211] X-ray powder diffraction analysis showed that the product was crystal form A.

[0212] Example 27: Preparation of Crystal Form A

[0213] 10 mg of compound 1 was weighed, 0.1 ml of ethyl acetate was added and stirred at room temperature for dissolution, 0.4 ml of n-heptane was added and stirred for 1 day, and then the product was evaporated and crystallized to obtain the product.

[0214] X-ray powder diffraction analysis showed that the product was crystal form A.

[0215] Example 28: Preparation of Crystal Form A

[0216] 10 mg of compound 1 was weighed, 0.1 ml of methyl tert-butyl ether was added and stirred at room temperature to dissolve, 0.1 ml of cyclohexane was added to precipitate, 0.2 ml of cyclohexane was added and stirred for 1 day, and the solid was filtered and vacuum dried to obtain the product.

[0217] X-ray powder diffraction analysis showed that the product was crystal form A.

[0218] Example 29: Preparation of Crystal Form A

[0219] 10 mg of compound 1 was weighed, 0.1 ml of tetrahydrofuran was added and stirred at room temperature to dissolve, 0.4 ml of cyclohexane was added and stirred for 1 day, and then evaporated and crystallized to obtain the product.

[0220] X-ray powder diffraction analysis showed that the product was crystal form A.

[0221] Example 30: Preparation of Crystal Form A

[0222] 10 mg of compound 1 was weighed, 0.1 ml of methyl isobutyl ketone was added and stirred at room temperature for dissolution, 0.4 ml of cyclohexane was added and stirred for 1 day, and then evaporated and crystallized to obtain the product.

[0223] X-ray powder diffraction analysis showed that the product was crystal form A.

[0224] Example 31: Preparation of Crystal Form A

[0225] 10 mg of compound 1 was weighed, 0.1 ml of 10% water / acetone was added and stirred at room temperature to dissolve, 0.1 ml of purified water was added to precipitate, 0.2 ml of purified water was added and stirred for 1 day, and the solid was filtered and vacuum dried to obtain the product.

[0226] X-ray powder diffraction analysis showed that the product was crystal form A.

[0227] Example 32: Preparation of Crystal Form A

[0228] 10 mg of compound 1 was weighed, 0.1 ml of isopropyl acetate was added and dissolved by stirring at room temperature, 0.4 ml of cyclohexane was added, and the product was evaporated and crystallized to obtain the product.

[0229] X-ray powder diffraction analysis showed that the product was crystal form A.

[0230] Example 33: Preparation of Crystal Form B

[0231] 10 mg of compound 1 was weighed, 0.1 ml of dimethyl sulfoxide was added to dissolve, 0.2 ml of water was added to crystallize, and beating was continued for 1 day. After filtering, vacuum drying was performed to obtain a solid.

[0232] The product was defined as Form B by X-ray powder diffraction analysis. The XRPD spectrum is shown in FIG2 , and the positions of the characteristic peaks are shown in Table 6.

[0233] The DSC spectrum showed endothermic peaks at 81.99 and 173.56 °C.

[0234] The TGA spectrum showed that the weight loss from 30°C to 100°C was 18.35%.

[0235] Table 6

[0236] Example 34: Preparation of Crystal Form C

[0237] 30 mg of compound 1 was weighed and added to 1.2 ml of 1,4-dioxane, stirred and dissolved, 1.8 ml of n-heptane was added, stirred and crystallized, filtered and vacuum dried to obtain a solid.

[0238] The product was defined as Form C by X-ray powder diffraction analysis. The XRPD spectrum is shown in FIG3 , and the positions of the characteristic peaks are shown in Table 7 .

[0239] The DSC spectrum showed endothermic peaks at 141.90 and 176.81 °C.

[0240] The TGA spectrum showed a weight loss of 1.89% from 30°C to 85°C and a weight loss of 5.94% from 85°C to 160°C.

[0241] Table 7

[0242] Example 35: Preparation of Crystal Form D

[0243] 30 mg of compound 1 was weighed, 0.3 ml of tetrahydrofuran was added and stirred at room temperature to dissolve, 1.2 ml of water was added and stirred to crystallize, the crystals were stirred and filtered, and vacuum dried to obtain a solid product.

[0244] The product was defined as D-type by X-ray powder diffraction analysis. The XRPD spectrum is shown in FIG4 , and the positions of the characteristic peaks are shown in Table 8.

[0245] The DSC spectrum showed an endothermic peak at 176.85°C.

[0246] The TGA spectrum showed a weight loss of 0.34% from 30°C to 100°C.

[0247] Table 8

[0248] Example 36: Preparation of Form E

[0249] 100 mg of compound 1 was weighed and dispersed in 1 mL of acetonitrile, stirred and slurried for 48 hours, filtered, and the solid was collected and dried in vacuo at 45° C. to obtain a solid product.

[0250] The product was defined as Form E by X-ray powder diffraction analysis. The X-ray powder diffraction data are shown in Table 9, and the X-ray powder diffraction spectrum is shown in FIG5 .

[0251] The DSC spectrum shows that the endothermic peaks are 88.25℃ and 177.28℃.

[0252] The TGA spectrum showed that the compound lost 1.80% of its weight from 30°C to 115°C, and lost 5.29% of its weight from 115°C to 230°C.

[0253] Table 9

[0254] Example 37: Preparation of Form F

[0255] 10 mg of compound 1 was weighed, 0.4 ml of 1,4-dioxane was added to dissolve the solution, 0.4 ml of n-heptane was added and stirred for 1 day for crystallization, 0.2 ml of n-heptane was added and stirred for 3 hours, and the supernatant was evaporated and crystallized to obtain a solid product.

[0256] The product was defined as Form F by X-ray powder diffraction analysis. The XRPD spectrum is shown in FIG6 , and the positions of the characteristic peaks are shown in Table 10.

[0257] Table 10

[0258] Test Case 2: Influencing Factors

[0259] The A crystal form was exposed and laid flat, and the stability of the sample was investigated under light (4500 Lux), high temperature (40° C., 60° C.), and high humidity (RH 75%, RH 92.5%) conditions for 30 days.

[0260] Table 11 Factors affecting crystal stability

[0261] Conclusion: The influencing factor experiment shows that the physical and chemical stability of free A crystal form is good under light, high temperature of 40℃ and 60℃, and high humidity of 75% and 92.5% for 30 days.

[0262] Test Example 3: Long-term accelerated test

[0263] The stability of Form A was investigated under conditions of 25°C / 60% RH and 40°C / 75% RH.

[0264] Table 12 Long-term accelerated stability of crystal form

[0265] Conclusion: Long-term accelerated experiments show that the physical and chemical stability of Form A is good at 25°C / 60% RH and 40°C / 75% RH for 6 months.

Claims

1. Crystalline Form A 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 7.237, 9.232, 13.702, 14.459 and 18.917, preferably has characteristic peaks at 7.237, 9.232, 13.702, 14.459, 18.917, 24.428 and 29.321, more preferably has characteristic peaks at 7.237, 9.232, 13.702, 14.459, 18.033, 18.917, 24.428, 25.521 and 29.321, and most preferably the X-ray powder diffraction pattern expressed in terms of a diffraction angle of 2θ is shown in Figure 1.

2. Crystalline Form B 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 8.205, 9.781, 12.870, 15.907 and 19.796, preferably has characteristic peaks at 8.205, 9.781, 12.870, 15.907, 19.448, 19.796, 20.264 and 23.185, more preferably has characteristic peaks at 8.205, 9.781, 10.672, 12.870, 15.356, 15.907, 16.997, 19.448, 19.796, 20.264 and 23.185, and most preferably has an X-ray powder diffraction pattern as shown in Figure 2.

3. Crystalline Form C 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 9.296, 15.522, 18.784, 23.216 and 25.889, preferably has characteristic peaks at 7.753, 9.296, 15.522, 18.784, 21.398, 23.216 and 25.889, more preferably has characteristic peaks at 7.353, 7.753, 9.296, 14.475, 15.522, 16.248, 17.320, 18.784, 21.398, 23.216 and 25.889, and most preferably has an X-ray powder diffraction pattern as shown in Figure 3.

4. Crystalline Form D 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 7.350, 12.084, 15.384, 18.643 and 29.312, preferably has characteristic peaks at 7.350, 12.084, 15.384, 16.260, 17.855, 18.643, 21.610 and 29.312, more preferably has characteristic peaks at 7.350, 12.084, 15.384, 16.260, 17.855, 18.643, 21.610, 22.768, 24.347, 25.201, 26.038 and 29.312, and most preferably the X-ray powder diffraction pattern expressed as a diffraction angle 2θ is shown in Figure 4.

5. Crystalline Form E 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 13.489, 18.000, 23.559, 24.276 and 26.328, preferably at 13.489, 16.863, 18.000, 23.559, 24.276, 26.108, 26.328 and 27.094, more preferably at 13.489, 14.587, 16.863, 18.000, 18.943, 23.279, 23.559, 24.276, 25.768, 26.108, 26.328 and 27.094, and most preferably the X-ray powder diffraction pattern expressed as a diffraction angle 2θ is shown in Figure 5.

6. Crystalline Form F 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 9.299, 14.439, 15.621, 16.200 and 17.314, preferably has characteristic peaks at 7.734, 9.299, 14.439, 15.621, 16.200, 17.314, 21.395 and 25.814, more preferably has characteristic peaks at 7.734, 9.299, 14.439, 15.621, 16.200, 17.314, 21.055, 21.395, 23.194, 25.814, 28.906, 34.485 and 43.544, and most preferably the X-ray powder diffraction pattern, expressed as a diffraction angle of 2θ, is shown in Figure 6.

7. The crystal form according to any one of claims 1 to 6, characterized in that The error range of the 2θ value is ±0.

2.

8. A method for preparing the crystal form according to any one of claims 1 to 7, selected from the following methods: Method 1: (a) mixing the compound 8-chloro-N-(2,2-difluorobenzo[d][1,3]dioxolan-5-yl)quinolin-2-amine with a solvent, stirring and dissolving the mixture or heating the mixture to dissolve the mixture; (b) adding a second solvent and crystallizing; Or, method 2: (a) mixing the compound 8-chloro-N-(2,2-difluorobenzo[d][1,3]dioxolan-5-yl)quinolin-2-amine with a solvent, stirring and dissolving the mixture or heating the mixture to dissolve the mixture; (b) crystallization; Or, method three: (a) mixing the compound 8-chloro-N-(2,2-difluorobenzo[d][1,3]dioxolan-5-yl)quinolin-2-amine with a solvent, (b) Stirring and beating.

9. A pharmaceutical composition comprising the crystal form according to any one of claims 1 to 7, and a pharmaceutically acceptable excipient.

10. A pharmaceutical composition prepared from the crystal form according to any one of claims 1 to 7 and a pharmaceutically acceptable excipient.

11. Use of the crystalline form according to any one of claims 1 to 7, or the pharmaceutical composition according to claim 9 or 10, 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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