Crystalline forms of quinoline amine compounds and their preparation methods

Abstract

The present disclosure relates to crystalline forms of quinolinamine compounds and preparation methods thereof. Specifically, the present disclosure provides a crystalline form of 8-chloro-N-(2,2-difluorobenzo[d][1,3]dioxolan-5-yl)quinolin-2-amine and preparation methods thereof, and the corresponding crystalline form has good stability and can be used in clinical treatment.

Description

[Technical Field]

[0001] This application claims priority from Chinese Patent Application No. 2022114917970, filed on November 25, 2022. The entire text of the above Chinese patent application is incorporated herein by reference.

[0002] The present disclosure belongs to the pharmaceutical field and relates to crystalline forms of quinolinamine compounds and methods for preparing same. [Background technology]

[0003] miR-124 is widely expressed in tissues throughout the body, with particularly high expression in brain tissue. Research has shown that overexpression of miR-124 promotes the transition of activated macrophages and microglia to a quiescent state, thereby inhibiting the autoimmune disease encephalomyelitis. Furthermore, miR-124 promotes the conversion of macrophages to the M2 type, thereby exerting anti-inflammatory effects. miR-124 also affects T cell differentiation, and miR-124-treated T cells exhibit reduced levels of both IFN-γ and TNFα. Overexpression of miR-124 exerts anti-inflammatory effects by downregulating STAT3 protein, reducing the expression of the pro-inflammatory cytokine IL-17, and inhibiting the differentiation of Th17 cells. These studies suggest that upregulating miR-124 could lead to the development of novel small molecule drugs that can be used to effectively treat related inflammatory diseases.

[0004] Related published patent applications include WO2010143169A2, WO2015001518A1, WO2016009065A2, WO2017158201A1, and WO2020127843A1, etc.

[0005] WO2022247920 discloses quinolinamine compounds that can upregulate miR-124, the structures of which are as follows: [ka]

[0006] The crystalline structure of a pharmaceutical active ingredient often affects the chemical and physical stability of the drug, and depending on the crystallization conditions and storage conditions, the crystalline structure of the compound may change, and may also result in the formation of other crystalline forms.Generally, amorphous drug products do not have a regular crystalline structure, and often have other defects such as relatively poor product stability, relatively difficult filtration, prone to solidification, poor flowability, etc.Therefore, studying its crystalline form is of great significance for the development of drugs suitable for industrial production and with good biological activity. [Prior art documents] [Patent documents]

[0007] [Patent Document 1] WO2010143169A2 [Patent Document 2] WO2015001518A1 [Patent Document 3] WO2016009065A2 [Patent Document 4] WO2017158201A1 [Patent Document 5] WO2020127843A1 [Patent Document 6] WO2022247920 Summary of the Invention

[0008] In one embodiment of the present disclosure, there is provided a crystalline form A of the compound 8-chloro-N-(2,2-difluorobenzo[d][1,3]dioxolan-5-yl)quinolin-2-amine, which has a powder X-ray diffraction pattern expressed in 2θ angles having characteristic peaks at 7.237, 9.232, 13.702, 14.459, and 18.917.

[0009] In some embodiments, the above type A crystal has characteristic peaks at 7.237, 9.232, 13.702, 14.459, 18.917, 24.428 and 29.321 in the powder X-ray diffraction pattern represented by the diffraction angle 2θ.

[0010] In some embodiments, the above type A crystal has characteristic peaks at 7.237, 9.232, 13.702, 14.459, 18.033, 18.917, 24.428, 25.521 and 29.321 in the powder X-ray diffraction pattern represented by the diffraction angle 2θ.

[0011] In some other embodiments, the above type A crystal has a powder X-ray diffraction pattern represented by the diffraction angle 2θ as shown in FIG. 1.

[0012] The present disclosure further provides a method for preparing the type A crystal of the above compound, and the above method is selected from any one of the following methods: Method 1: (a) Mix the compound 8-chloro-N-(2,2-difluorobenzo[d][1,3]dioxolan-5-yl)quinolin-2-amine with solvent (1), and stir to dissolve it or heat to dissolve it. (b) Add solvent (2) and perform crystallization. Among them, 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. Or, Method 2: (a) Mix the compound 8-chloro-N-(2,2-difluorobenzo[d][1,3]dioxolan-5-yl)quinolin-2-amine with solvent (3), and stir to dissolve it or heat to dissolve it. (b) Perform crystallization. Among them, solvent (3) is selected from tetrahydrofuran, ethyl acetate or dichloromethane. Or, Method 3: (a) mixing the compound 8-chloro-N-(2,2-difluorobenzo[d][1,3]dioxolan-5-yl)quinolin-2-amine with a solvent (4); (b) stirring to form a slurry, wherein 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;

[0013] In one embodiment of the present disclosure, there is provided a B-form crystal of the compound 8-chloro-N-(2,2-difluorobenzo[d][1,3]dioxolan-5-yl)quinolin-2-amine, which has a powder X-ray diffraction pattern expressed in 2θ angles having characteristic peaks at 8.205, 9.781, 12.87, 15.907, and 19.796.

[0014] In some embodiments, the B-type crystal has a powder X-ray diffraction pattern expressed in terms of diffraction angles 2θ, which has characteristic peaks at 8.205, 9.781, 12.870, 15.907, 19.448, 19.796, 20.264, and 23.185.

[0015] In some embodiments, the type B crystal has a powder X-ray diffraction pattern expressed in terms of diffraction angles 2θ, which has 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.

[0016] In some other embodiments, the powder X-ray diffraction pattern of the B-type crystals, expressed in terms of diffraction angle 2θ angles, is as shown in FIG.

[0017] The present disclosure further provides a method for preparing crystalline Form B of the compound, the method comprising: (a) mixing the compound 8-chloro-N-(2,2-difluorobenzo[d][1,3]dioxolan-5-yl)quinolin-2-amine with a solvent (5) and dissolving the compound by stirring or heating; (b) adding solvent (6) and crystallizing; Among them, solvent (5) is selected from dimethyl sulfoxide, and solvent (6) is selected from water.

[0018] In one embodiment of the present disclosure, there is provided a C-form crystal of the compound 8-chloro-N-(2,2-difluorobenzo[d][1,3]dioxolan-5-yl)quinolin-2-amine, which has a powder X-ray diffraction pattern expressed in 2θ angles having characteristic peaks at 9.296, 15.522, 18.784, 23.216, and 25.889.

[0019] In some embodiments, the C-type crystal has a powder X-ray diffraction pattern expressed in terms of diffraction angle 2θ angles, which has characteristic peaks at 7.753, 9.296, 15.522, 18.784, 21.398, 23.216, and 25.889.

[0020] In some embodiments, the C-type crystal has a powder X-ray diffraction pattern expressed in terms of diffraction angles 2θ, which 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.

[0021] In some other embodiments, the powder X-ray diffraction pattern of the C-type crystals, expressed in terms of diffraction angle 2θ angles, is as shown in FIG.

[0022] The present disclosure further provides a method for preparing crystalline Form C of the compound, the method comprising: (a) mixing the compound 8-chloro-N-(2,2-difluorobenzo[d][1,3]dioxolan-5-yl)quinolin-2-amine with a solvent (7) and dissolving it by stirring or heating; (b) adding solvent (8) and crystallizing; Among them, solvent (7) is selected from 1,4-dioxane, and solvent (8) is selected from n-heptane.

[0023] In one embodiment of the present disclosure, there is provided a D-form crystal of the compound 8-chloro-N-(2,2-difluorobenzo[d][1,3]dioxolan-5-yl)quinolin-2-amine, which has a powder X-ray diffraction pattern expressed in 2θ angles having characteristic peaks at 7.350, 12.084, 15.384, 18.643, and 29.312.

[0024] In some embodiments, the D-type crystal has a powder X-ray diffraction pattern expressed in terms of diffraction angles 2θ, which has characteristic peaks at 7.350, 12.084, 15.384, 16.260, 17.855, 18.643, 21.610, and 29.312.

[0025] In some embodiments, the D-type crystal has a powder X-ray diffraction pattern expressed in terms of diffraction angles 2θ, which 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.

[0026] In some other embodiments, the D-type crystal has a powder X-ray diffraction pattern, expressed in terms of diffraction angle 2θ, as shown in FIG.

[0027] The present disclosure further provides a method for preparing crystalline Form D of the compound, the method comprising: (a) mixing the compound 8-chloro-N-(2,2-difluorobenzo[d][1,3]dioxolan-5-yl)quinolin-2-amine with a solvent (9) and dissolving the compound by stirring or heating; (b) adding solvent (10) and crystallizing; Among them, solvent (9) is selected from tetrahydrofuran, and solvent (10) is selected from water.

[0028] In one embodiment of the present disclosure, there is provided a crystalline form E of the compound 8-chloro-N-(2,2-difluorobenzo[d][1,3]dioxolan-5-yl)quinolin-2-amine, which has a powder X-ray diffraction pattern expressed in 2θ angles having characteristic peaks at 13.489, 18.000, 23.559, 24.276, and 26.328.

[0029] In some embodiments, the E-form crystal has a powder X-ray diffraction pattern expressed in terms of diffraction angles 2θ, which has characteristic peaks at 13.489, 16.863, 18.000, 23.559, 24.276, 26.108, 26.328, and 27.094.

[0030] In some embodiments, the E-form crystal has a powder X-ray diffraction pattern expressed in terms of diffraction angles 2θ, which has 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.

[0031] In some other embodiments, the powder X-ray diffraction pattern of the E-form crystals, expressed in terms of diffraction angle 2θ angles, is as shown in FIG.

[0032] In one embodiment of the present disclosure, there is provided a crystalline form F of the compound 8-chloro-N-(2,2-difluorobenzo[d][1,3]dioxolan-5-yl)quinolin-2-amine, which has a powder X-ray diffraction pattern expressed in 2θ angles having characteristic peaks at 9.299, 14.439, 15.621, 16.200, and 17.314.

[0033] In some embodiments, the F-type crystal has a powder X-ray diffraction pattern expressed in terms of diffraction angles 2θ, which has characteristic peaks at 7.734, 9.299, 14.439, 15.621, 16.200, 17.314, 21.395, and 25.814.

[0034] In some embodiments, the F-type crystal has a powder X-ray diffraction pattern expressed in terms of diffraction angles 2θ, which 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.

[0035] In some other embodiments, the F-type crystal has a powder X-ray diffraction pattern, expressed in terms of diffraction angle 2θ, as shown in FIG.

[0036] Furthermore, the compound A-type crystals, B-type crystals, C-type crystals, D-type crystals, E-type crystals, or F-type crystals described in the present disclosure have a powder X-ray diffraction pattern expressed in terms of diffraction angle 2θ angles, and the error range of the 2θ angles is ±0.2.

[0037] 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.

[0038] In some embodiments, the crystallization includes, but is not limited to, agitation crystallization (elution crystallization, slurry crystallization) and volatilization crystallization.

[0039] In some embodiments, the drying method includes, but is not limited to, air drying and vacuum drying. The drying temperature is generally 25°C to 100°C, preferably 30°C to 70°C, for example, 40°C, 50°C, or 60°C.

[0040] In another aspect, the present disclosure further provides a pharmaceutical composition comprising the crystalline form and a pharmaceutically acceptable excipient.

[0041] The present disclosure further provides a pharmaceutical composition prepared with the above crystalline form and a pharmaceutically acceptable excipient.

[0042] The present disclosure further provides a method for preparing a pharmaceutical composition comprising mixing the crystalline form with a pharmaceutically acceptable excipient.

[0043] The present disclosure further provides a use of the crystalline form or pharmaceutical composition in the preparation of a medicament for modulating miRNA levels, preferably wherein the miRNA is miR-124.

[0044] The present disclosure further provides the use of said crystalline form or pharmaceutical composition in a medicament for treating and / or preventing a disease or condition, wherein said disease or condition is selected from inflammation and cancer.

[0045] In some embodiments, the inflammation is inflammatory bowel disease, hi some embodiments, the cancer is melanoma or breast cancer.

[0046] The term "2θ or 2θ angle" used in the present disclosure refers to the diffraction angle, where θ is the Bragg angle and is expressed in ° or degrees. The error range of 2θ for each characteristic peak is ±0.20 (including rounding to the nearest decimal place), and specifically includes -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.

[0047] "Differential scanning calorimetry or DSC" as used herein refers to the measurement of temperature and heat flow differences between a sample and a reference during a heating or isothermal process of the sample to characterize all physical and chemical changes associated with thermal effects and obtain information on the phase transitions of the sample.

[0048] The drying temperature described in the present disclosure is generally 25°C to 100°C, preferably 30°C to 70°C, and drying may be performed under normal pressure or reduced pressure.

[0049] "Pharmaceutically acceptable excipients" as referred to in this disclosure include, but are not limited to, any auxiliary agent, carrier, glidant, sweetener, diluent, preservative, dye / colorant, flavoring agent, surfactant, wetting agent, dispersing agent, suspending agent, stabilizer, isotonic agent, or emulsifier that has already been approved by the U.S. Food and Drug Administration and is acceptable for use in humans or domestic animals.

[0050] The term "slurrying" used in this disclosure refers to a purification method that utilizes the properties of a substance having poor solubility in a solvent but good solubility of impurities in the solvent, and purification by slurrying can decolorize, change the crystal form, or remove small amounts of impurities.

[0051] The crystalline forms described in this disclosure include, but are not limited to, solvates of compounds of Formula (I), including, but not limited to, solvates of compounds of Formula (I), including, but not limited to, water. [Brief explanation of the drawings]

[0052] [Figure 1] 1 is an XRPD spectrum of the A-type crystal of Compound 1. [Figure 2] 1 is an XRPD spectrum of type B crystal of Compound 1. [Figure 3] 1 is an XRPD spectrum of the C-type crystal of Compound 1. [Figure 4] 1 is an XRPD spectrum of the D-type crystal of Compound 1. [Figure 5] 1 is an XRPD spectrum of E-form crystals of Compound 1. [Figure 6] 1 is an XRPD spectrum of the F-type crystal of Compound 1. DETAILED DESCRIPTION OF THE INVENTION

[0053] The present disclosure is further illustrated by the following examples and experimental examples, which are for illustrative purposes only and are not intended to limit the scope of the disclosure.

[0054] Test conditions for the equipment used in the experiment: The structure of the compound is determined by nuclear magnetic resonance (NMR) and / or mass spectrometry (MS). The NMR shifts (δ) are 10 -6 The NMR data are shown in ppm. A Bruker AVANCE-400 nuclear magnetic resonance spectrometer was used for the NMR measurements. The solvents used were deuterated dimethyl sulfoxide (DMSO-d6), deuterated chloroform (CDCl3), and deuterated methanol (CD3OD). The internal standard was tetramethylsilane (TMS).

[0055] For MS measurements, a liquid chromatograph mass spectrometer, Agilent 1200 / 1290 DAD-6110 / 6120 Quadrupole MS (manufacturer: Agilent, MS model number: 6110 / 6120 Quadrupole MS), a waters ACQuity UPLC-QD / SQD (manufacturer: waters, MS model number: waters ACQuity Qda Detector / waters SQ Detector), and a THERMO Ultimate 3000-Q Exactive (manufacturer: THERMO, MS model number: THERMO Q 15 Exactive) were used.

[0056] For the HPLC measurements, a high performance liquid chromatograph Agilent 1260DAD (Sunfire C18 150 x 4.6 mm column) and a high performance liquid chromatograph Thermo U3000 (Gimini C18 150 x 4.6 mm column) were used.

[0057] XRPD is a method of detecting powder X-ray diffraction. A BRUKER D8 X-ray diffractometer is used for the measurement. The specific collected information is a Cu anode (40 kV, 40 mA), radiation: monochromatic Cu-Ka radiation (l = 1.5418 Å), scanning method: θ / 2θ, scanning range: 3 to 48 o is.

[0058] DSC stands for differential scanning calorimetry. A METTLER TOLEDO DSC 3+ differential scanning calorimeter was used for the measurement, with a heating rate of 10°C / min, 25 to 300°C, or 25 to 350°C, and a nitrogen gas purge rate of 50 mL / min.

[0059] TGA is thermogravimetric analysis. A METTLER TOLEDO TGA 2 type thermogravimetric analyzer is used for detection. The heating rate is 10°C / min. The specific temperature range is based on the corresponding pattern. The nitrogen gas purge rate is 50mL / min.

[0060] DVS stands for dynamic moisture sorption. Instru- mental Surface Measurement Systems were used. The humidity range was considered from 0% to 95% in 10% increments, starting from 50%. The criterion was a mass change per gradient dM / dT ≤ 0.002%, with a TMAX of 360 min, and two cycles were repeated.

[0061] Known starting materials of the present disclosure may be synthesized by or according to methods known in the art, or may be purchased from companies such as ABCR GmbH & Co. KG, Acros Organics, Aldrich Chemical Company, Accela ChemBio Inc., and Darui Chemicals. In the examples, thin layer chromatography (TLC) was used to monitor the reaction process. The developing solvents used in the reactions, the column chromatographic eluent system used to purify the compounds, and the developing solvent system for thin layer chromatography included A: dichloromethane / methanol system and B: n-hexane / ethyl acetate system. The volume ratio of the solvents may be adjusted according to the polarity of the compounds, and may be adjusted by adding small amounts of basic or acidic reagents such as triethylamine and acetic acid.

[0062] Example 1. Synthesis of 8-chloro-N-(2,2-difluorobenzo[d][1,3]dioxolan-5-yl)quino-2-amine (see preparation method of Example 1 in application number WO2022247920) [ka]

[0063] 2,8-Dichloroquinoline 1a (100 mg, 0.51 mmol, BiDe Pharmaceutical) 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), heated to 90 °C, and reacted for 12 hours. The reaction mixture was filtered and then purified by high-performance liquid preparative separation (Waters 2767-SQ Detecor2, elution system: 0.1% aqueous formic acid and acetonitrile, acetonitrile gradient: 65% to 85%, flow rate: 30 mL / min) to obtain the title compound 1 (150 mg, 89.0% yield). MS m / z (ESI): 335.0 [M+1]. 1 H NMR (500 MHz, DMSO-d6) δ 9.99 (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).

[0064] Test Example 1: Upregulation effect on miR-124 1. Experimental materials and equipment 1. Human T cell activator CD3 / CD28 magnetic beads (Dynabead Human T-Activator CD3 / CD28 for T Cell Expansion and Activation) (Gibco, 11131D) 2. Human Total T Cell Isolation Kit (Pan T Cell Isolation Kit, human) (Miltenyi, 130-096-535) 3. Human Interleukin 2 (IL-2) (Peprotech, 200-02-100) 4. MicroRNA Extraction Reagent Kit (Qiagen, 217004) 5. Mini RNA Reverse Transcription Kit (miScript II RT Kit) (Qiagen, 218161) 6. Small RNA SYBR Green PCR Reagent Kit (miScript SYBR Green PCR Kit) (Qiagen, 218073) 7. Phosphate buffer PBS, pH 7.4 (Shanghai Yuanpei Biotechnology Co., Ltd., B320) 8. Bovine serum albumin, BSA (Hekidonten, ST023) 9.EDTA (0.5M), pH 8.0 (Invitrogen, AM9260G) 10. LS Columns (Miltenyi, 130-042-401) 11. 24-well cell culture plate (Corning, 3524) 12. 96-well plate (Corning, 3788) 13. Cell incubator (Thermo, Steri cycle i160) 14. Real-time fluorescent quantitative PCR instrument (Applied Biosystems, QuantStudio6 Flex) 15. PCR device (Applied biosystem, ProFlex) 16. 96-well clear PCR plate, 0.2 mL (Applied biosystems, N8010560) 17. RPMI 1640 medium (Gibco, 11875119) 18. Fetal bovine serum, FBS (Gibco, 10099-141) 19. Magnetic rack (Invitrogen, DynaMag TM -2) 20. 6-well cell culture plate (Thermo, 150239) 21.Spectrophotometer (IMPLEN, NP80) 22. Magnetic Bead Separator Rack (QuadroMACS Separator) (Miltenyi, 130-090-976) 23. miR124-3P-F primer (customized by GENEWIZ) 24. hsa-U6 detection primer (TIANGEN, CD201-0145)

[0065] 2. Experimental Procedure The effects of compounds on miR-124 expression levels were detected in T cells after CD3 / CD28 antibody activation. After treatment of activated T cells with compounds, total cellular RNA was extracted, and the resulting cDNA was reverse-transcribed using specific miR-124 primers and quantified by SYBR green fluorescent quantitative PCR.

[0066] Isolation of T cells: Human peripheral blood mononuclear cells (PBMCs) were purchased, counted, filtered, washed once with isolation buffer (PBS pH 7.4, containing 0.5% BSA and 2 mM EDTA), the supernatant was discarded, and 1 × 10 cells were collected. 7 Each component was added to the cells in an amount of 40 μL of buffer and 10 μL of pan T cell biotin-antibody cocktail per cell, and the precipitate was resuspended and mixed uniformly. The mixture was then incubated in a refrigerator at 4°C for 5 minutes. After incubation, 1 × 10 cells were collected. 7 Each component was added at a ratio of 30 μL of buffer solution and 20 μL of T cell separation magnetic beads (Pan T Cell MicroBeads Cocktail) per cell, mixed uniformly, and then incubated in a refrigerator at 4°C for 10 minutes. The LS column was pre-rinsed with 3 mL of cell separation buffer, and the cell suspension was applied to the column. After applying the cell suspension to the column, the column was washed three times with 1 mL of cell separation buffer. The effluent cell solution was collected in a 15 mL centrifuge tube, i.e., enriched T cells. The cells were counted and counted at 1 x 10 6 The cells were cultured at a density of 10 cells / mL in RPMI1640 medium (complete medium) containing 10% FBS and 40 U / mL IL-2, and stored on ice until ready for use.

[0067] T cell activation: 1×10 6Add 25 μL of activated magnetic beads per cell. Remove the corresponding T cell-activating CD3 / CD28 magnetic beads and place them in a 1.5 mL centrifuge tube. Shake on a shaker for approximately 30 seconds before aspirating. Wash the activated magnetic beads three times with medium in a volume ratio of greater than 1:1 in the centrifuge tube. Finally, remove all wash solution and resuspend the activated magnetic beads in an equal volume of complete medium. Add the washed activated magnetic beads to the cell resuspension and mix thoroughly. Remove the 6-well plate, add 3 mL of cells per well, and culture in a cell incubator at 37°C and 5% CO2 for two days.

[0068] Compound treatment: 20 mM compound stock solution was diluted with DMSO to 200 μM, and then further diluted 4-fold with complete medium to 50 μM (50x), mixed uniformly, and prepared for use. A 4-fold dilution with DMSO (25% DMSO) was used as a negative control. T cells were activated for 2 days, and then the cells were pipetted uniformly. A 1.5 mL centrifuge tube was attached using a magnetic rack, the activated magnetic beads were removed, and the cell suspension was collected. After counting the cells, they were centrifuged at 300 × g for 10 minutes, the supernatant was discarded, and 1.02 × 10 cells were collected. 6 980 μL of cell suspension and 20 μL of 50× compound were added to each 24-well plate to give a final compound concentration of 1 μM. Cells were cultured in a 37°C, 5% CO2 cell incubator for 3 days.

[0069] RNA extraction: T cells were collected by centrifugation, filtered at 1500 rpm for 3 minutes, washed once with PBS, and centrifuged. The supernatant was discarded. Total RNA was extracted from the cells using a small RNA extraction reagent kit according to the manufacturer's instructions. 700 μL of Trizol cell lysis solution was added to the cell pellet, pipetted evenly with a pipette head, and allowed to stand at room temperature for 5 minutes. 140 μL of chloroform was added, mixed evenly by shaking, and allowed to stand at room temperature for 3 minutes. The chloroform-cell lysis solution mixture was centrifuged at 12,000 × g for 15 minutes at 4°C. The upper layer was transferred to a new RNase-free centrifuge tube, and 1.5 volumes of absolute ethanol were added. The mixture was pipetted several times with a pipette head. The solution was transferred to an RNA adsorption column and centrifuged at 8,000 × g for 15 seconds. The centrifugal column was washed once with 700 μL of RWT solution and centrifuged at 8,000 × g for 15 s. Then, 500 μL of RPE solution was added and washed twice, centrifuged at 8,000 × g for 2 min. The adsorption column was placed in a new 2 mL centrifuge tube and centrifuged at 12,000 × g for 1 min to remove residual washing solution. The adsorption column was placed in a new 1.5 mL centrifuge tube, 30–50 μL of RNase-free water was added, and centrifuged at 12,000 × g for 2 min. The collected solution was used as the RNA solution, and the RNA concentration was measured using a spectrophotometer. The RNA solution was stored in a -80°C refrigerator.

[0070] Reverse transcription: The extracted RNA template was placed on ice, the mini RNA reverse transcription kit was removed, and some of the components (including 5x miScript HiSpec Buffer, 10x miScript nucleic acid mix, and RNA enzyme-free water) were thawed at room temperature. The miScript Reverse Transcriptase mix components were thawed on ice. Each reaction (10 μL) consisted of 5x miScript HiSpec Buffer (2 μL), 10x miScript nucleic acid mix (1 μL), miScript Reverse Transcriptase mix (1 μL), RNA enzyme-free water (2 μL), and RNA template (4 μL). The reaction was prepared on ice. The sample was placed in a PCR machine and the program was set as follows: 37°C for 60 minutes, 95°C for 5 minutes, and stored at 4°C. The completed reaction sample became the cDNA sample.

[0071] Fluorescent quantitative PCR: The transcription level of miR-124 was detected using SYBR green staining, and the transcription level of the housekeeping gene U6 was detected as an internal standard. All reagents required for the small RNA SYBR green PCR reagent kit were thawed to room temperature, and each cDNA sample template was diluted 10-fold with RNA enzyme-free water, followed by a further 5-fold dilution. The reaction mixture was prepared according to Table 1 below, placed in a 96-well PCR plate, blocked with blocking film, and centrifuged. PCR reactions were performed using a fluorescent quantitative PCR instrument according to the steps in Table 2.

[0072] [Table 1]

[0073] [Table 2]

[0074] [Table 3]

[0075] Data analysis: The C values ​​calculated by the software were used to calculate the ratio of the expression level of miR-124 in each sample to the internal standard U6, i.e., ΔC (test compound) = C miRNA-124 (Test Compound)-CT U6 (test compound) was calculated. The relative expression level was calculated as follows: relative expression level (test compound) = 2 (-[ΔCT(試験化合物)-ΔCT(DMSO)]) It was calculated by the formula:

[0076] The miR-124 upregulation (fold) of Compound 1 was 3.9-fold, and it has good activity in promoting the upregulation of miR124.

[0077] Example 2: Preparation of type A crystals 250 mg of compound 1 was taken and added to 2.5 mL of acetonitrile with stirring 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 dried under vacuum to obtain a solid.

[0078] When detected by powder X-ray diffraction, the product was defined as type A crystal, and the powder X-ray diffraction data was shown in Table 4, and the powder X-ray diffraction spectrum was shown in Figure 1.

[0079] The DSC spectrum showed an endothermic peak at 177.14°C.

[0080] The TGA spectrum showed a weight loss of 0.22% between 30°C and 100°C.

[0081] DVS detection showed that the moisture weight gain of the sample was about 0.02% under normal storage conditions (i.e., room temperature, 60% RH), about 0.02% under accelerated storage conditions (i.e., 70% RH), and about 0.18% under extreme storage conditions (i.e., 90% RH). Furthermore, re-measurement of the crystal form after DVS detection showed no change in the crystal form.

[0082] [Table 4]

[0083] Example 3: Preparation of type A crystals Compound 1 (6 mg, 17.63 μmol) was dissolved in 0.3 mL of tetrahydrofuran, clarified, and then slowly evaporated to give a solid, which was dried under vacuum at 45° C. for 3 hours to give the product.

[0084] When detected by powder X-ray diffraction, the product was found to be type A crystal.

[0085] Example 4: Preparation of type A crystals Compound 1 (11 mg, 32.87 μmol) was dissolved in 0.4 mL of ethyl acetate, clarified, and then slowly evaporated to give a solid, which was dried under vacuum at 45° C. for 3 hours to give the product.

[0086] When detected by powder X-ray diffraction, the product was found to be type A crystal.

[0087] Example 5: Preparation of type A crystals Compound 1 (14 mg, 41.83 μmol) was dissolved in 0.5 mL of dichloromethane, clarified, and then slowly evaporated to give a solid, which was dried under vacuum at 45° C. for 3 hours to give the product.

[0088] When detected by powder X-ray diffraction, the product was found to be type A crystal.

[0089] Example 6: Preparation of type A crystals Compound 1 (1.70 g, 5.08 mmol) was dispersed in 35 mL of cyclohexane, slurried with stirring for 48 hours, filtered to collect the filter cake, and dried under vacuum at 45° C. for 16 hours to obtain the product.

[0090] When detected by powder X-ray diffraction, the product was found to be type A crystal.

[0091] Example 7: Preparation of type A crystals 10 mg of Compound 1 was weighed out, and 1 mL of purified water was added thereto, followed by slurrying at room temperature for 3 days. After filtration, the solid was dried under vacuum to obtain the product.

[0092] When detected by powder X-ray diffraction, the product was found to be type A crystal.

[0093] Example 8: Preparation of type A crystals Referring to the method of Example 7, 10 mg of Compound 1 was weighed out, added to the solvent, slurried, filtered, and the solid was dried under vacuum to obtain the product.

[0094] The product was found to be a type A crystal by powder X-ray diffraction, and the data are shown in Table 5.

[0095] [Table 5]

[0096] Example 9: Preparation of type A crystals 10 mg of compound 1 was weighed, 0.2 mL of methanol was added, and the mixture was stirred at room temperature to clarify the mixture, 0.2 mL of purified water was added, and the mixture was stirred to crystallize the mixture, and the mixture was slurried at room temperature for 1 day. After filtration, the solid was dried under vacuum to obtain the product.

[0097] When detected by powder X-ray diffraction, the product was found to be type A crystal.

[0098] Example 10: Preparation of type A crystals 10 mg of Compound 1 was weighed, 0.2 mL of ethanol was added, and the mixture was stirred at room temperature to clarify the mixture, 0.4 mL of n-heptane was added, and the mixture was stirred for 1 day, followed by evaporation and crystallization to obtain the product.

[0099] When detected by powder X-ray diffraction, the product was found to be type A crystal.

[0100] Example 11: Preparation of type A crystals 10 mg of Compound 1 was weighed, 0.6 mL of isopropanol was added, and the mixture was stirred at room temperature to clarify the mixture, 0.4 mL of n-heptane was added, and the mixture was stirred for 1 day, followed by evaporation and crystallization to obtain the product.

[0101] When detected by powder X-ray diffraction, the product was found to be type A crystal.

[0102] Example 12: Preparation of type A crystals 10 mg of Compound 1 was weighed, 0.1 mL of acetone was added, and the mixture was stirred at room temperature to clarify the mixture, 0.4 mL of n-heptane was added, and the mixture was stirred for 1 day, followed by evaporation and crystallization to obtain the product.

[0103] When detected by powder X-ray diffraction, the product was found to be type A crystal.

[0104] Example 13: Preparation of type A crystals 10 mg of Compound 1 was weighed, 0.1 mL of ethyl acetate was added, and the mixture was stirred at room temperature to clarify the mixture, 0.4 mL of n-heptane was added, and the mixture was stirred for 1 day, followed by evaporation and crystallization to obtain the product.

[0105] When detected by powder X-ray diffraction, the product was found to be type A crystal.

[0106] Example 14: Preparation of type A crystals 10 mg of compound 1 was weighed, 0.1 mL of isopropyl acetate was added, and the mixture was stirred at room temperature to clarify the mixture. 0.4 mL of n-heptane was added, and the mixture was stirred to crystallize the mixture. The mixture was then slurried at room temperature for 1 day, supplemented with 0.2 mL of n-heptane, and the mixture was further slurried for 3 hours. After filtration, the solid was dried under vacuum to obtain the product.

[0107] When detected by powder X-ray diffraction, the product was found to be type A crystal.

[0108] Example 15: Preparation of type A crystals 10 mg of compound 1 was weighed, 0.1 mL of tetrahydrofuran was added, and the mixture was stirred at room temperature to clarify the mixture, 0.4 mL of n-heptane was added, and the mixture was stirred for 1 day, followed by evaporation and crystallization to obtain the product.

[0109] When detected by powder X-ray diffraction, the product was found to be type A crystal.

[0110] Example 16: Preparation of type A crystals 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 clarify the mixture, 0.4 mL of n-heptane was added, and the mixture was stirred for 1 day, followed by evaporation and crystallization to obtain the product.

[0111] When detected by powder X-ray diffraction, the product was found to be type A crystal.

[0112] Example 17: Preparation of type A crystals 10 mg of compound 1 was weighed, 0.4 mL of dichloromethane was added, and the mixture was stirred at room temperature to clarify the crystallization. 0.4 mL of n-heptane was added to the mixture, and the mixture was slurried at room temperature for 1 day. The mixture was supplemented with 0.2 mL of n-heptane and slurried for 3 hours. After filtration, the solid was dried under vacuum to obtain the product.

[0113] When detected by powder X-ray diffraction, the product was found to be type A crystal.

[0114] Example 18: Preparation of type A crystals 10 mg of compound 1 was weighed, 1 mL of 10% water / methanol was added, and the mixture was stirred at room temperature to clarify the crystallization. 0.1 mL of water was added, and the mixture was stirred to crystallize the crystals. 0.1 mL of water was added, and the mixture was subsequently slurried for 1 day. After filtration, the solid was dried under vacuum to obtain the product.

[0115] When detected by powder X-ray diffraction, the product was found to be type A crystal.

[0116] Example 19: Preparation of type A crystals 10 mg of Compound 1 was weighed, and 0.6 mL of 7% water / ethanol was added thereto, followed by stirring at room temperature to clarify the mixture. 0.4 mL of n-heptane was added thereto, and the mixture was evaporated to crystallize the mixture, thereby obtaining the product.

[0117] When detected by powder X-ray diffraction, the product was found to be type A crystal.

[0118] Example 20: Preparation of type A crystals 10 mg of Compound 1 was weighed, and 0.6 mL of 10% water / isopropanol was added thereto, followed by stirring at room temperature to clarify the mixture, and 0.4 mL of n-heptane was added thereto, followed by evaporation to crystallize the mixture, thereby obtaining the product.

[0119] When detected by powder X-ray diffraction, the product was found to be type A crystal.

[0120] Example 21: Preparation of type A crystals 10 mg of Compound 1 was weighed, and 0.1 mL of 10% water / acetone was added thereto, followed by stirring at room temperature to clarify the mixture. 0.4 mL of n-heptane was added thereto, and the mixture was evaporated to crystallize the mixture, thereby obtaining the product.

[0121] When detected by powder X-ray diffraction, the product was found to be type A crystal.

[0122] Example 22: Preparation of type A crystals 10 mg of compound 1 was weighed, 0.2 mL of ethanol was added, and the mixture was stirred at room temperature to dissolve. 0.1 mL of purified water was added, and the mixture was stirred to precipitate. 0.2 mL of purified water was then added, and the mixture was stirred for 1 day. After filtration, the solid was dried under vacuum to obtain the product.

[0123] When detected by powder X-ray diffraction, the product was found to be type A crystal.

[0124] Example 23: Preparation of type A crystals 10 mg of Compound 1 was weighed, 0.2 mL of ethanol was added, and the mixture was stirred at room temperature to dissolve. 0.4 mL of cyclohexane was added, and the mixture was stirred for 1 day, after which the mixture was evaporated and crystallized to obtain the product.

[0125] When detected by powder X-ray diffraction, the product was found to be type A crystal.

[0126] Example 24: Preparation of type A crystals 10 mg of compound 1 was weighed, 0.1 mL of acetone was added, and the mixture was stirred at room temperature to dissolve. 0.1 mL of purified water was added, and the mixture was stirred to precipitate. 0.2 mL of purified water was then added, and the mixture was stirred for 1 day. After filtration, the solid was dried under vacuum to obtain the product.

[0127] When detected by powder X-ray diffraction, the product was found to be type A crystal.

[0128] Example 25: Preparation of type A crystals 10 mg of Compound 1 was weighed, 0.1 mL of acetone was added, and the mixture was stirred at room temperature to dissolve. 0.4 mL of cyclohexane was added, and the mixture was stirred for 1 day, after which the mixture was evaporated and crystallized to obtain the product.

[0129] When detected by powder X-ray diffraction, the product was found to be type A crystal.

[0130] Example 26: Preparation of A-type crystals 10 mg of Compound 1 was weighed, 0.1 mL of ethyl acetate was added, and the mixture was stirred at room temperature to dissolve. 0.4 mL of cyclohexane was added, and the mixture was stirred for 1 day, after which evaporation and crystallization were carried out to obtain the product.

[0131] When detected by powder X-ray diffraction, the product was found to be type A crystal.

[0132] Example 27: Preparation of type A crystals 10 mg of Compound 1 was weighed, 0.1 mL of ethyl acetate 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, after which evaporation and crystallization were carried out to obtain the product.

[0133] When detected by powder X-ray diffraction, the product was found to be type A crystal.

[0134] Example 28: Preparation of type A crystals 10 mg of compound 1 was weighed, 0.1 mL of methyl tert-butyl ether was added, and the mixture was stirred at room temperature to dissolve. 0.1 mL of cyclohexane was added to precipitate the compound, and then 0.2 mL of cyclohexane was added. The mixture was stirred for 1 day, filtered, and the solid was dried under vacuum to obtain the product.

[0135] When detected by powder X-ray diffraction, the product was found to be type A crystal.

[0136] Example 29: Preparation of type A crystals 10 mg of Compound 1 was weighed, 0.1 mL of tetrahydrofuran was added, and the mixture was stirred at room temperature to dissolve. 0.4 mL of cyclohexane was added, and the mixture was stirred for 1 day, after which evaporation and crystallization were carried out to obtain the product.

[0137] When detected by powder X-ray diffraction, the product was found to be type A crystal.

[0138] Example 30: Preparation of type A crystals 10 mg of Compound 1 was weighed, 0.1 mL of methyl isobutyl ketone was added, and the mixture was stirred at room temperature to dissolve. 0.4 mL of cyclohexane was added, and the mixture was stirred for 1 day, after which evaporation and crystallization were carried out to obtain the product.

[0139] When detected by powder X-ray diffraction, the product was found to be type A crystal.

[0140] Example 31: Preparation of type A crystals 10 mg of compound 1 was weighed, and 0.1 mL of 10% water / acetone was added. The mixture was stirred at room temperature to dissolve the compound, and 0.1 mL of purified water was added to cause precipitation. 0.2 mL of purified water was then added, and the mixture was stirred for 1 day. After filtration, the solid was dried under vacuum to obtain the product.

[0141] When detected by powder X-ray diffraction, the product was found to be type A crystal.

[0142] Example 32: Preparation of type A crystals 10 mg of Compound 1 was weighed, and 0.1 mL of isopropyl acetate was added thereto. The mixture was stirred at room temperature to dissolve the compound, and 0.4 mL of cyclohexane was added thereto, followed by evaporation and crystallization to obtain a product.

[0143] When detected by powder X-ray diffraction, the product was found to be type A crystal.

[0144] Example 33: Preparation of B-type crystals 10 mg of Compound 1 was weighed out, clarified by adding 0.1 mL of dimethyl sulfoxide, crystallized by adding 0.2 mL of water, slurried for 1 day, filtered, and dried under vacuum to obtain a solid.

[0145] When detected by powder X-ray diffraction, the product was defined as B-type crystals, and its XRPD spectrum is shown in FIG. 2, and its characteristic peak positions are shown in Table 6.

[0146] The DSC spectrum showed endothermic peak values ​​at 81.99 and 173.56°C.

[0147] The TGA spectrum showed a weight loss of 18.35% between 30 and 100 °C.

[0148] [Table 6]

[0149] Example 34: Preparation of C-type crystals 30 mg of Compound 1 was weighed, added to 1.2 mL of 1,4-dioxane, and stirred to dissolve. 1.8 mL of n-heptane was added, and stirred to cause crystallization. The mixture was filtered and then dried under vacuum to obtain a solid.

[0150] When detected by powder X-ray diffraction, the product was defined as C-type crystals, and its XRPD spectrum is shown in FIG. 3, and its characteristic peak positions are shown in Table 7.

[0151] The DSC spectrum showed endothermic peak values ​​at 141.90 and 176.81°C.

[0152] 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.

[0153] [Table 7]

[0154] Example 35: Preparation of D-type crystals 30 mg of compound 1 was weighed, 0.3 mL of tetrahydrofuran was added, and the mixture was stirred at room temperature to dissolve. 1.2 mL of water was added, and the mixture was stirred to crystallize. The mixture was filtered and then dried under vacuum to obtain a solid product.

[0155] When detected by powder X-ray diffraction, the product was defined as D-type crystal, and its XRPD spectrum is shown in Figure 4, and its characteristic peak positions are shown in Table 8.

[0156] The DSC spectrum showed an endothermic peak at 176.85°C.

[0157] The TGA spectrum showed a weight loss of 0.34% between 30°C and 100°C.

[0158] [Table 8]

[0159] Example 36: Preparation of E-type crystals 100 mg of compound 1 was weighed and dispersed in 1 mL of acetonitrile, stirred for 48 hours to form a slurry, filtered, and then the solid was collected and dried under vacuum at 45° C. to obtain a product solid.

[0160] The product was identified as E-type crystals by powder X-ray diffraction. The powder X-ray diffraction data is shown in Table 9, and the powder X-ray diffraction spectrum is shown in Figure 5.

[0161] The DSC spectrum showed endothermic peak values ​​at 88.25°C and 177.28°C.

[0162] The TGA spectrum showed a weight loss of 1.80% of the compound between 30°C and 115°C, and a weight loss of 5.29% of the compound between 115°C and 230°C.

[0163] [Table 9]

[0164] Example 37: Preparation of F-type crystals 10 mg of compound 1 was weighed out, clarified with 0.4 mL of 1,4-dioxane, added with 0.4 mL of n-heptane, stirred for 1 day to allow crystallization, supplemented with 0.2 mL of n-heptane, slurried for 3 hours, filtered, and then evaporated to obtain the crystallized product solid.

[0165] When detected by powder X-ray diffraction, the product was defined as F-type crystals, and its XRPD spectrum is shown in FIG. 6, and its characteristic peak positions are shown in Table 10.

[0166] [Table 10]

[0167] Test Example 2: Influencing Factors The A-type crystals were placed flat in an open state, and the stability of the samples was examined under conditions of light exposure (4500 Lux), high temperature (40°C, 60°C), and high humidity (RH 75%, RH 92.5%). The sampling period was 30 days.

[0168] [Table 11]

[0169] Conclusion: Experiments on influencing factors showed that the physical and chemical stability of free A-type crystals was good under conditions of light irradiation, high temperatures of 40℃ and 60℃, and high humidity of 75% and 92.5% for 30 days.

[0170] Test example 3: Long-term accelerated test The stability of the A-type crystals was examined under the conditions of 25°C / 60% RH and 40°C / 75% RH.

[0171] [Table 12]

[0172] Conclusion: The long-term accelerated experiment shows that the physical and chemical stability of the A-type crystals is good for 6 months under the conditions of 25°C / 60%RH and 40°C / 75%RH.

Claims

1. A type A crystal of the compound 8-chloro-N-(2,2-difluorobenzo[d][1,3]dioxolan-5-yl)quinolin-2-amine, characterized in that the powder X-ray diffraction pattern expressed in terms of diffraction angle 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 has a powder X-ray diffraction pattern expressed in terms of diffraction angle 2θ as shown in FIG. A type crystal.

2. The B-type crystal of the compound 8-chloro-N-(2,2-difluorobenzo[d][1,3]dioxolan-5-yl)quinolin-2-amine has a powder X-ray diffraction pattern expressed in terms of diffraction angle 2θ angles, which has characteristic peaks at 8.205, 9.781, 12.870, 15.907, 19.448, and 19.796, and preferably has characteristic peaks at 8.205, 9.781, 12.870, 15.907, 19.448, and 19. and more preferably, it 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, it 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 its powder X-ray diffraction pattern, expressed in terms of diffraction angle 2θ angles, is as shown in FIG.

2. B type crystal.

3. The C-type crystal of the compound 8-chloro-N-(2,2-difluorobenzo[d][1,3]dioxolan-5-yl)quinolin-2-amine has characteristic peaks at 9.296, 15.522, 18.784, 23.216 and 25.889 in a powder X-ray diffraction pattern expressed as a diffraction angle 2θ angle, and preferably has characteristic peaks at 7.753, 9.296, 15.522, 18.784, 21.39 and more preferably, it 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 ... a powder X-ray diffraction pattern expressed in terms of diffraction angle 2θ angles as shown in FIG. C type crystal.

4. The D-type crystal of the compound 8-chloro-N-(2,2-difluorobenzo[d][1,3]dioxolan-5-yl)quinolin-2-amine has a powder X-ray diffraction pattern represented by a diffraction angle 2θ angle having characteristic peaks at 7.350, 12.084, 15.384, 18.643 and 29.312, preferably 7.350, 12.084, 15.384, 16.260, 17.855, 18.

643. , 21.610 and 29.312, more preferably 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 characterized in that the powder X-ray diffraction pattern, expressed in terms of diffraction angle 2θ angles, is as shown in FIG. D type crystal.

5. The E-form crystal of the compound 8-chloro-N-(2,2-difluorobenzo[d][1,3]dioxolan-5-yl)quinolin-2-amine has characteristic peaks at 13.489, 18.000, 23.559, 24.276, and 26.328 in a powder X-ray diffraction pattern expressed as a diffraction angle 2θ angle, and preferably has characteristic peaks at 13.489, 16.863, 18.000, 23.559, 24.276, 26.10 and more preferably, it has 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, and most ... a powder X-ray diffraction pattern represented in terms of diffraction angle 2θ angles as shown in FIG. E type crystal.

6. The F-type crystal of the compound 8-chloro-N-(2,2-difluorobenzo[d][1,3]dioxolan-5-yl)quinolin-2-amine has characteristic peaks at 9.299, 14.439, 15.621, 16.200, and 17.314 in a powder X-ray diffraction pattern expressed as a diffraction angle 2θ angle, and preferably has characteristic peaks at 7.734, 9.299, 14.439, 15.621, 16.200, 17.314, 21. and 25.814, more preferably 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 characterized in that the powder X-ray diffraction pattern, expressed in terms of diffraction angle 2θ, is as shown in FIG. F type crystal.

7. The error range of the 2θ value is ±0.

2. The crystalline form according to any one of claims 1 to 6.

8. A method for preparing the crystalline form according to any one of claims 1 to 7, comprising any one of the following methods: Method one: (a) mixing the compound 8-chloro-N-(2,2-difluorobenzo[d][1,3]dioxolan-5-yl)quinolin-2-amine with a solvent and dissolving it by stirring or heating; (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 and dissolving it by stirring or heating; (b) crystallizing; Or, Method 3: (a) mixing the compound 8-chloro-N-(2,2-difluorobenzo[d][1,3]dioxolan-5-yl)quinolin-2-amine with a solvent; (b) stirring to form a slurry; Preparation method.

9. A pharmaceutical composition comprising the crystalline form of any one of claims 1 to 7 and a pharmaceutically acceptable excipient. Pharmaceutical compositions.

10. Prepared by the crystalline form of any one of claims 1 to 7 and a pharmaceutically acceptable excipient. Pharmaceutical compositions.

11. 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 said disease or condition is selected from inflammation and cancer, said inflammation preferably being selected from inflammatory bowel disease and said cancer preferably being selected from melanoma or breast cancer. use.

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