Crystalline form of maleic acid salt of polysubstituted benzene ring compound, its preparation method and use

Abstract

The present invention discloses a crystalline form of a maleic acid salt of a polysubstituted benzene ring compound, its preparation method and use. The crystalline form of the salt of a polysubstituted benzene ring compound represented by Formula II of the present invention has high physicochemical stability, high solubility, low moisture absorption, high flowability, more options for formulation processes, and is more suitable for industrial production, which is of great value for the optimization and development of pharmaceuticals. JPEG2023545165000031.jpg79107

Description

[Technical Field]

[0001] This application claims priority from Chinese Patent Application No. 2020110887642, filed on October 13, 2020. The entire text of the above Chinese patent application is incorporated herein by reference. The present invention relates to a crystalline form of a maleate salt of a polysubstituted benzene ring compound, a method for preparing the same, and uses thereof. [Background technology]

[0002] A polysubstituted benzene ring compound, the structure of which is shown in Formula I.

[0003] [ka]

[0004] The compound disclosed in Patent WO2020020374A1 is an EZH2 (Enhancer of Zeste Homolog 2) inhibitor and can be used for the prevention or treatment of EZH2-mediated diseases, including brain cancer, thyroid cancer, cardiac sarcoma, lung cancer, oral cancer, gastric cancer and various other cancers.

[0005] The phenomenon that a substance exists in two or more different crystalline structures is called polymorphism. For pharmaceuticals, such polymorphism may affect the absorption and bioavailability of the pharmaceutical, leading to different clinical therapeutic effects and toxic side effects. In view of this, it is of great significance to develop a superior crystalline form of the polysubstituted benzene ring compound salt represented by Formula I with excellent performance.

[0006] During the research and development of pharmaceuticals, both solubility and hygroscopicity are factors that need to be considered, but there is usually a certain correlation between the two indicators, and since active ingredients that are soluble in water themselves can also absorb moisture, substances with high solubility are usually prone to hygroscopicity. Therefore, the development of compounds with high solubility and low hygroscopicity is of great significance to the research and development of pharmaceuticals. Summary of the Invention [Problem to be solved by the invention]

[0007] The technical problem to be solved by the present invention is to provide a crystalline form of a salt of a polysubstituted benzene ring compound represented by Formula II, its preparation method and use thereof, in order to overcome the defects of the prior art that the polysubstituted benzene ring compound represented by Formula I has low solubility and is easily hygroscopic during pharmaceutical preparation. This crystalline form has high physicochemical stability, high solubility, low hygroscopicity, and good flowability, allowing for more formulation process options and being more suitable for industrialized production, and is of great value to the optimization and development of pharmaceuticals. [Means for solving the problem]

[0008] The present invention provides crystalline form A of the poly-substituted benzene ring compound maleate salt represented by formula II, which has an X-ray powder diffraction pattern shown in 2θ angles at diffraction peaks at 7.39±0.2°, 8.78±0.2°, 13.99±0.2°, 15.21±0.2°, 15.73±0.2°, 17.75±0.2°, 18.51±0.2°, and 21.06±0.2°.

[0009] [ka]

[0010] The crystalline form A of the poly-substituted benzene ring compound maleate salt represented by formula II may have an X-ray powder diffraction pattern shown in 2θ angles, which may also have diffraction peaks at one or more 2θ angles of 8.51±0.2° and 16.06±0.2°.

[0011] The crystalline form A of the poly-substituted benzene ring compound maleate salt represented by formula II may have an X-ray powder diffraction pattern shown in 2θ angles, which may also have diffraction peaks at one or more 2θ angles of 18.20±0.2° and 20.68±0.2°.

[0012] The crystalline form A of the poly-substituted benzene ring compound maleate salt represented by formula II may have an X-ray powder diffraction pattern shown in 2θ angles, which may also have diffraction peaks at one or more 2θ angles of 22.81±0.2° and 25.28±0.2°.

[0013] The crystalline form A of the polysubstituted benzene ring compound maleate salt represented by the formula II may have an X-ray powder diffraction pattern shown at 2θ angles, its diffraction peaks and relative intensities shown in Table 1.

[0014] [Table 1-1]

[0015] [Table 1-2]

[0016] The crystalline form A of the polysubstituted benzene ring compound maleate salt represented by the formula II may have an X-ray powder diffraction pattern shown at 2θ angles essentially as shown in FIG.

[0017] In the present invention, the X-ray powder diffraction patterns were all measured using Kα radiation from a Cu target.

[0018] In the present invention, the crystalline form A of the polysubstituted benzene ring compound maleate salt represented by the formula II has major endothermic peaks at 185.3°C±3°C and 204.4°C±3°C in a differential scanning calorimetry (DSC) diagram.

[0019] In the present invention, the DSC of the crystalline form A of the poly-substituted benzene ring compound maleate salt represented by the formula II may be essentially as shown in FIG.

[0020] In the present invention, the thermogravimetric analysis diagram (TGA) of the crystalline form A of the poly-substituted benzene ring compound maleate salt represented by the formula II may be essentially as shown in FIG.

[0021] The present invention further provides a method for preparing crystalline form A of the poly-substituted benzene ring compound maleate salt of formula II, which is Method 1, Method 2, Method 3, Method 4, Method 5 or Method 6. Method 1: Reacting a polysubstituted benzene ring compound of formula I with maleic acid in solvent A; the solvent A is one or more selected from methyl isobutyl ketone, 2-methyltetrahydrofuran, and ethyl acetate; Method 2: Form B of the crystalline form of the maleate salt of the polysubstituted benzene ring compound represented by formula II is suspended in solvent B, followed by stirring. the solvent B is one or more selected from the group consisting of alcohol-based solvents, ester-based solvents, ketone-based solvents, ether-based solvents, benzene-based solvents, and nitrile-based solvents; Method 3: Crystalline Form B of the polysubstituted benzene ring compound maleate salt of Formula II is saturated in a mixed solvent of dimethyl sulfoxide and ethyl acetate, and then crystallized. Method 4: Crystalline Form B of the polysubstituted benzene ring compound maleate salt of Formula II is dissolved in Solvent C, and Solvent D is added to the solution to allow crystallization. the solvent C is one or more selected from methanol, dimethyl sulfoxide, and trichloromethane; when the solvent C is methanol, the solvent D is ethyl acetate; when the solvent C is dimethyl sulfoxide, the solvent D is isopropyl acetate; when the solvent C is trichloromethane, the solvent D is one or more of ethyl acetate, acetonitrile, and butanone; Method 5: A gas-liquid permeation crystallization experiment may be carried out on the crystalline form B of the polysubstituted benzene ring compound maleate salt represented by formula II, wherein the good solvent is methanol, and the anti-solvent is one or more selected from acetone, ethyl acetate, and tetrahydrofuran; Method 6: A gas-solid permeation experiment may be carried out on the crystalline form B of the polysubstituted benzene ring compound maleate salt of formula II in a solvent E environment; The solvent E is one or more of methanol, acetonitrile, acetone, dimethylsulfoxide, isopropanol, and dimethylformamide.

[0022] In the above methods 2 to 6, the crystalline form B of the polysubstituted benzene ring compound maleate salt represented by the formula II has an X-ray powder diffraction pattern shown at 2θ angles basically as shown in FIG.

[0023] In Method 1, the reaction temperature may be any temperature conventional in the art, and is preferably room temperature.

[0024] In Method 1, the reaction time may be a time commonly used in the art, and is preferably 2 to 4 days.

[0025] In Method 1, the molar ratio of the polysubstituted benzene ring compound represented by Formula I to the maleic acid may be a common ratio in the art, and is preferably 1:(1 to 1.2), for example, 1:1.07.

[0026] In Method 1, the volume / mass ratio of the solvent A to the polysubstituted benzene ring compound represented by Formula I may be a conventional ratio in the art, and is preferably 15 to 40 mL / g, for example, 17.6 mL / g.

[0027] In Method 2, the stirring may be conventional in the art, preferably magnetic stirring.

[0028] In Method 2, the stirring temperature may be a temperature commonly used in the art, preferably 0 to 50°C, for example, room temperature, 50°C, or 50°C to 5°C, and temperature cycling is performed under conditions of 0.1°C / min. When the stirring temperature is 50°C, the stirring time is 5 to 8 days, and when the stirring temperature is room temperature, the stirring time is 25 to 35 days. When the stirring temperature is temperature cycling, the temperature cycling conditions are 50°C to 5°C, 0.1°C / min, and 2 to 4 cycles.

[0029] In Method 2, the alcoholic solvent may be any one commonly used in the art, and is preferably ethanol and / or isopropanol.

[0030] In Method 2, the ester solvent may be any common one in the art, preferably ethyl acetate and / or isopropyl acetate.

[0031] In Method 2, the ketone solvent may be any conventional one in the art, and is preferably one or more of acetone, butanone, and methyl isobutyl ketone.

[0032] In Method 2, the ethereal solvent may be any common one in the art, preferably one or more of anisole, cyclopentyl methyl ether and methyl t-butyl ether, more preferably anisole and / or cyclopentyl methyl ether.

[0033] In Method 2, the benzene-based solvent may be any conventional one in the art, and is preferably toluene.

[0034] In Method 2, the nitrile solvent may be any conventional solvent in the art, and is preferably acetonitrile.

[0035] In Method 2, the volume / mass ratio of the solvent B to the crystalline form B of the poly-substituted benzene ring compound maleate salt represented by Formula II may be a volume / mass ratio common in the art, preferably 20 to 50 mL / g, for example, 25 mL / g.

[0036] In Method 3, the volume ratio of dimethyl sulfoxide to ethyl acetate in the mixed solvent may be a common ratio in the art, and is preferably 1:(2 to 5), for example, 1:3.

[0037] In Method 3, the volume / mass ratio of the mixed solvent to the crystalline form B of the poly-substituted benzene ring compound maleate salt represented by Formula II may be a volume / mass ratio common in the art, preferably 20 to 50 mL / g, for example, 25 mL / g.

[0038] In Method 4, the method of adding the solvent D may be a method commonly used in the art, preferably dropwise.

[0039] In Method 4, the volume ratio of the solvent D to the solvent C may be a common ratio in the art, and is preferably (7-15):1, for example, 9:1.

[0040] In Method 5, the temperature of the gas-liquid permeation experiment may be any temperature conventional in the art, and is preferably room temperature.

[0041] In Method 5, the volume ratio of the antisolvent to the good solvent may be a common ratio in the art, and is preferably (7-15):1, for example, 8:1.

[0042] In Method 6, the temperature of the gas-solid permeation experiment may be any temperature conventional in the art, preferably room temperature.

[0043] In Method 6, the time period for the gas-solid permeation experiment may be a period commonly used in the art, and is preferably 20 to 50 days, for example, 30 days.

[0044] In Method 6, the volume / mass ratio of the solvent E to the crystalline form B of the poly-substituted benzene ring compound maleate salt of Formula II may be a volume / mass ratio common in the art, preferably 100 to 400 mL / g, for example, 200 mL / g.

[0045] In the methods 3 to 5, the solution preparation method may refer to a conventional preparation method in the art, for example, a method for preparing a saturated solution is adopted, and the saturated solution is preferably filtered, preferably filtered through a filtration membrane;

[0046] In the above methods 3 to 5, the crystallization may be a conventional method for such operations in the art.

[0047] For example, in Method 3, the crystallization method is preferably a slow temperature decrease, and the slow temperature decrease method is preferably from 50°C to 5°C, and the slow temperature decrease rate is 0.1 to 0.5°C / min, preferably 0.1°C / min, and if the solution is still clear, it is transferred to room temperature and evaporated.

[0048] For example, in Method 4, preferably, after adding solvent D, a solid precipitates; if no solid precipitates, the temperature is lowered to 5°C; if no solid precipitates again, the mixture is evaporated at room temperature or vacuum dried at 50°C.

[0049] For example, in Method 5, the crystallization method is preferably room temperature crystallization or vacuum drying at 50°C.

[0050] The above methods 1 to 6 may further include post-treatment steps of separation and drying. The separation method may be a method conventional in the art, preferably centrifugation or filtration. The drying method may be a method conventional in the art, preferably vacuum drying.

[0051] Preferably, the method for preparing crystalline form B of the poly-substituted benzene ring compound maleate salt represented by formula II includes the steps of dissolving the poly-substituted benzene ring compound represented by formula I and maleic acid in ethanol, lowering the temperature to cause crystallization, and adding methyl t-butyl ether.

[0052] In the method for preparing crystalline form B of the polysubstituted benzene ring compound maleate salt of formula II, the molar ratio of the polysubstituted benzene ring compound of formula I to the maleic acid may be a molar ratio commonly used in the art, and is preferably 1:(0.95-0.98), for example, 1:0.978.

[0053] In the method for preparing crystalline form B of the polysubstituted benzene ring compound maleate salt represented by formula II, the mass ratio of the ethanol to the polysubstituted benzene ring compound represented by formula I may be a mass ratio common in the art, preferably (7-9):1, for example, 6.9:1.

[0054] In the method for preparing crystalline form B of the polysubstituted benzene ring compound maleate salt represented by formula II, the temperature-lowering crystallization may be performed at a temperature commonly used in the art, preferably 15 to 25°C, for example, 20°C.

[0055] In the method for preparing crystalline form B of the polysubstituted benzene ring compound maleate salt represented by formula II, the mass ratio of the methyl t-butyl ether to the polysubstituted benzene ring compound represented by formula I may be a mass ratio common in the art, preferably (12-15):1, for example, 13:1.

[0056] The method for preparing crystalline form B of the poly-substituted benzene ring compound maleate salt of formula II may further include post-treatment steps of filtration and drying, where the filtration may be carried out under the usual conditions and operations for such operations in the art. Preferably, the filter cake obtained after filtration is further washed with methyl t-butyl ether, and the drying may be carried out under the usual conditions and operations for such operations in the art.

[0057] The present invention further provides a pharmaceutical composition comprising crystalline form A of the poly-substituted benzene ring compound maleate salt represented by formula II and a pharmaceutically acceptable carrier.

[0058] Crystalline Form A of the polysubstituted benzene ring compound maleate salt represented by Formula II according to the present invention or the pharmaceutical composition can be used to prevent or treat EZH2-mediated diseases. Preferably, the EZH2-mediated diseases include cancer, pulmonary arterial hypertension, myelofibrosis, human immunodeficiency virus (HIV) disease, graft-versus-host disease (GVHD), Weaver syndrome, psoriasis, and liver fibrosis. More preferably, the EZH2-mediated disease is cancer.

[0059] Preferably, the cancer comprises a metastatic or malignant tumor.

[0060] Preferably, the cancer includes brain cancer, thyroid cancer, cardiac sarcoma, lung cancer, oral cancer, gastric cancer, liver cancer, kidney cancer, pancreatic cancer, esophageal cancer, nasopharyngeal cancer, laryngeal cancer, colorectal cancer, breast cancer, prostate cancer, bladder cancer, ovarian cancer, uterine cancer, bone cancer, melanoma, glioblastoma, lymphoma, blood cancer, adrenal neuroblastoma, skin cancer, astrocytoma, etc.

[0061] In the present invention, crystalline form A of the poly-substituted benzene ring compound maleate salt represented by formula II can also be used in combination with one or more other active ingredients. When used in combination, the active ingredients may be in separate compositions, administered simultaneously or at different times by the same or different administration routes during treatment, or they may be administered together in the same pharmaceutical composition.

[0062] In the present invention, there is no particular limitation on the method of administration of the pharmaceutical composition, and various dosage forms, such as tablets, pills, solutions, suspensions, emulsions, granules, or capsules, can be selected and administered orally depending on the age, sex, other conditions, and symptoms of the patient; injections can be administered alone or mixed with an injection transport solution (e.g., glucose solution or amino acid solution) and then injected intravenously; and suppositories can be administered rectally.

[0063] In some instances, the crystalline form A of the polysubstituted benzene ring compound maleate salt represented by formula II is not converted when preparing a formulation with one or more pharmaceutically acceptable carriers and / or excipients and / or diluents.

[0064] In the present invention, room temperature is 10 to 35°C.

[0065] In the present invention, the water activity of the solvent used is a w <0.2.

[0066] In the present invention, "prophylactic treatment" refers to "prevention." "Prevention" refers to reducing the risk of suffering from or developing a disease or disorder (i.e., preventing at least one clinical symptom of the disease from occurring in a subject who may have been exposed to a disease-inducing agent or who is susceptible to the disease prior to the onset of the disease).

[0067] In the present invention, "treatment" refers to ameliorating a disease or disorder (i.e., preventing or eliminating the disease, reducing the extent or severity of its clinical symptoms), or improving at least one physical parameter, which may not be perceptible to the subject, or slowing the progression of the disease.

[0068] The crystalline forms of the present invention can be identified by one or more solid-state analytical methods, such as X-ray powder diffraction, single-crystal X-ray diffraction, infrared absorption spectroscopy, differential scanning calorimetry, and thermogravimetry. Those skilled in the art are aware that peak intensities and / or peak states in X-ray powder diffraction may vary depending on experimental conditions. Furthermore, due to differences in instrument precision, measured 2θ values ​​have an error of approximately ±0.2°. Because the relative peak intensity values ​​depend more on the characteristics of the measured sample, such as crystal size and purity, than on the peak position, measured peak intensities may vary by approximately ±20%. Despite experimental error, instrument error, and orientation preference, those skilled in the art can obtain sufficient information from the X-ray powder diffraction data of the patent to identify individual crystalline forms. In infrared spectroscopy, differences in the performance of various instrument models, as well as differences in the degree of polishing and water absorption during sample preparation, can have some effect on the spectral shape and absorption peak position. In DSC measurements, the initial temperature of the endothermic peak, the maximum temperature, and the heat of fusion data actually obtained all have a certain degree of variability depending on the heating rate, crystal shape and purity, and other measurement parameters.

[0069] Each of the above preferred conditions can be combined arbitrarily without departing from the common knowledge in the art to obtain each preferred embodiment of the present invention.

[0070] All of the reagents and raw materials used in the present invention are commercially available. [Effects of the Invention]

[0071] The positive advances of the present invention include a simple preparation method for crystalline Form A of the polysubstituted benzene ring compound maleate salt of Formula II according to the present invention, high physicochemical stability (no transformation of crystalline form or loss of chemical purity occurs even when left uncovered at 60°C for one day, or when left uncovered at 25°C / 60%RH or 40°C / 75%RH for one week), high solubility (solubility reaches 6 mg / mL in water and SGF, and 4 mg / mL in FaSSIF and FeSSIF), low hygroscopicity (water adsorption is 0.4% at 25°C / 80%RH, almost no hygroscopicity), and high fluidity (when D90 is controlled at 200 μm, the angle of repose of a single API salt is around 40°), which is more advantageous for the development of formulation processes for herbal medicines and has great value. [Brief explanation of the drawings]

[0072] [Figure 1] 1 is an X-ray powder diffraction pattern of crystalline form A of the polysubstituted benzene ring compound maleate salt represented by formula II. [Figure 2] 1 is a differential scanning calorimetry (DSC) diagram of crystalline form A of the maleate salt of a polysubstituted benzene ring compound represented by formula II. [Figure 3] 1 is a thermogravimetric analysis diagram (TGA) of crystalline form A of the maleate salt of a polysubstituted benzene ring compound represented by formula II. [Figure 4] 1 is an X-ray powder diffraction pattern of crystalline form B of the polysubstituted benzene ring compound maleate salt represented by formula II. [Figure 5] 1 is an X-ray powder diffraction pattern of the free crystalline form A of the polysubstituted benzene ring compound of formula I. [Figure 6] 1 is an X-ray powder diffraction pattern of the free crystalline form B of the polysubstituted benzene ring compound of formula I. [Figure 7] 1 is an X-ray powder diffraction pattern of crystalline form C of the polysubstituted benzene ring compound maleate salt represented by formula II. [Figure 8] 1 is an X-ray powder diffraction pattern of crystalline form D of the maleate salt of a polysubstituted benzene ring compound represented by formula II. [Figure 9] 1 is an X-ray powder diffraction pattern of crystalline form E of the maleate salt of a polysubstituted benzene ring compound represented by formula II. [Figure 10] 1 is an X-ray powder diffraction pattern of crystalline form A of the polysubstituted benzene ring compound succinate salt represented by formula III. [Figure 11] 1 is an X-ray powder diffraction pattern of crystalline form A of the hydrobromide salt of a polysubstituted benzene ring compound represented by formula IV. [Figure 12] 1 is an X-ray powder diffraction pattern of the free crystalline form D of the polysubstituted benzene ring compound of formula I. [Figure 13] 1 is an X-ray powder diffraction pattern of the free crystalline form C of the polysubstituted benzene ring compound of formula I. [Figure 14] 1 is an X-ray powder diffraction pattern of the free crystalline form E of the polysubstituted benzene ring compound of formula I. [Figure 15] 1 is an X-ray powder diffraction pattern of the free crystalline form F of the polysubstituted benzene ring compound of formula I. [Figure 16] 1 is an X-ray powder diffraction pattern of the free crystalline form G of the polysubstituted benzene ring compound of formula I. [Figure 17] 1 is an X-ray powder diffraction pattern of the free crystalline form H of the polysubstituted benzene ring compound of formula I. [Figure 18] 1 is an X-ray powder diffraction pattern of the free crystalline form I of the polysubstituted benzene ring compound of formula I. DETAILED DESCRIPTION OF THE INVENTION

[0073] The present invention will be further explained by the following examples, but the present invention is not limited to the scope of the described examples. In the following examples, experimental methods without specific conditions are selected according to conventional methods and conditions or product instructions. [Example]

[0074] The equipment used in the following examples is shown in Table 2.

[0075] [Table 2]

[0076] X-ray powder diffraction analysis (XRPD): XRPD patterns were collected on Empyrean and PANalytical X'Pert 3-ray powder diffraction analyzers, and the scanning parameters are listed in Table 3.

[0077] [Table 3]

[0078] Thermogravimetric analysis (TGA) and differential scanning calorimetry (DSC): TGA and DSC patterns were collected on a Discovery 5500 thermogravimetric analyzer and a Discovery 2500 differential scanning calorimeter, respectively, and the parameters are listed in Table 4.

[0079] [Table 4]

[0080] Dynamic Water Sorption (DVS) Dynamic moisture sorption (DVS) curves were collected on a DVS Intrinsic from Surface Measurement Systems (SMS). Relative humidity at 25 °C was corrected for the deliquescence points of LiCl, Mg(NO3)2, and KCl. DVS test parameters are listed in Table 5.

[0081] [Table 5]

[0082] Hydrogen spectrum liquid state nuclear magnetism ( 1 H Solution NMR Hydrogen Spectra Liquid-state nuclear magnetic spectra were collected on a Bruker 400M nuclear magnetic resonance instrument, with DMSO-d6 as the solvent.

[0083] Ultra-Performance Liquid Chromatography (UPLC) The purity and solubility of the samples under study were tested by Waters Ultra High Performance Liquid Chromatography, and the conditions are shown in Table 6.

[0084] [Table 6]

[0085] Example 1: Preparation of Crystalline Form A of the Polysubstituted Benzene Ring Compound of Formula I (See WO2020 / 020374A1) To a 10 mL DMF solution of 3-aminomethyl-4-methyl-6-methylpyridin-2(1H)-one hydrochloride (1 g, 5.3 mmol, 1.05 eq), EDC·HCl (1.2 g, 6.6 mmol, 1.25 eq), HOBt·HO (0.97 g, 6.6 mmol, 1.25 eq), and 3-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-2-methyl-5-(trans-3(piperidin-1-yl)cyclobutoxy)benzoic acid (2.1 g, 5.0 mmol, 1 eq) was added DIPEA (1.9 g, 15.1 mmol, 3 eq) at room temperature. After the reaction was completed, the mixture was stirred for 80 minutes. Add 1 mL of water, filter, and bake the filter cake to obtain 1.7 g of N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl-3-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-2-methyl-5-(trans-3(piperidin-1-yl)cyclobutoxy)benzamide. m / z (ES+), [M+H] + = 551.40; HPLC tR = 6.901 min. 1H NMR (400 MHz, DMSO-d6):δ11.45 (s, 1H),8.02 (t, J=4.8Hz, 1H), 6.59 (d, J=1.9Hz, 1H), 6.38 (d, J=2.0Hz, 1H), 5.85 (s, 1H), 4.70-4.63 (m, 1H), 4.25 (d, J=4.0Hz, 2H), 3.83 (d, J=12Hz, 2H), 3.24 (d, J=8Hz, 2H), 3.00-2.85 (m, 3H), 2.32-2.23 (m,10H), 2.10 (d, J=8Hz, 8H), 1.6-1.59 (m, 2H), 1.50 (m, 6H), 1.38 (s, 2H), 0.78 (t, J=8.0Hz, 1H).

[0086] Measurement of the X-ray powder diffraction spectrum showed that the obtained sample was the free crystalline form A of the polysubstituted benzene ring compound represented by formula I, and the X-ray powder diffraction pattern was shown in FIG.

[0087] Example 2: Preparation of crystalline form A of the polysubstituted benzene ring compound maleate salt represented by formula II 397.4 mg of crystalline form A of the polysubstituted benzene ring compound represented by formula I obtained in Example 1 was weighed into a 20 mL glass vial, 87.5 mg of maleic acid was added, and 7.0 mL of ethyl acetate was added to form a suspension, which was then magnetically stirred at room temperature for about 3 days. The resulting solid was separated by suction filtration and vacuum dried at room temperature overnight to obtain 443.8 mg of a solid sample.

[0088] Measurement of the X-ray powder diffraction spectrum revealed that the obtained sample was crystalline form A of the polysubstituted benzene ring compound maleate salt represented by formula II. The X-ray powder diffraction pattern shown in 2θ angles, its diffraction peaks, relative intensities, crystal plane spacing, peak heights and full width at half maximum (FWHM Left) may be shown in Table 7, and the X-ray powder diffraction pattern is shown in Figure 1.

[0089] [Table 7-1]

[0090] [Table 7-2]

[0091] In FIG. 2, the DSC spectrum showed that the crystalline form A of the polysubstituted benzene ring compound maleate salt represented by formula II exhibited endothermic signals at 185.3° C. and 204.4° C.

[0092] In FIG. 3, the TGA spectrum showed that the crystalline form A of the maleate salt of the polysubstituted benzene ring compound represented by formula II lost 1.58% weight at 160°C.

[0093] Crystalline form A of the polysubstituted benzene ring compound maleate salt represented by formula II 1 H NMR data are 1H NMR (400 MHz, DMSO-d6): δ 11.45 (s, 1H), 9.22 (brs, 1H), 8.03 (t, J = 4.8 Hz, 1H), 6.62 (d, J = 2.4Hz, 1H), 6.42 (d, J = 2.4Hz, 1H), 6.02 (s, 2H), 5.86 (s, 1H), 4.80 (t, J = 6.4Hz, 1H), 4.25 (d, J = 4.8Hz, 2H), 3.83 (d, J = 11.2Hz, 3H), 3.32-3.20 (m, 4H), 2.99 (dd, J = 6.8, 14.0Hz, 2H), 2.97-2.92 (m, 1H), 2.70-2.67 (m, 4H), 2.40-2.38 (m, 2H), 2.20 (s, 3H), 2.11(s, 3H), 2.10 (s, 1H), 1.81-1.46(m, 10H), 0.79 (t, J = 6.8Hz, 3H) ppm.

[0094] As a result, in crystalline form A of the maleate salt of the poly-substituted benzene ring compound represented by formula II, the molar ratio of the poly-substituted benzene ring compound represented by formula I to maleic acid is 1:1, the molar ratio of the free state of the poly-substituted benzene ring compound represented by formula I to ethyl acetate is 1:0.08, and the residual mass fraction of ethyl acetate is about 1.0%.

[0095] Example 3: Preparation of crystalline form A of the polysubstituted benzene ring compound maleate salt represented by formula II Compared with Example 2, only the type of solvent was different, and ethyl acetate in Example 2 was replaced with methyl isobutyl ketone and 2-methyltetrahydrofuran, respectively.

[0096] The X-ray powder diffraction spectrum of the sample obtained by this method was compared with the spectrum of the sample in Example 2, and both were identified as crystalline form A of the polysubstituted benzene ring compound maleate salt represented by formula II.

[0097] Example 4: Preparation of Crystalline Form B of the Maleate Salt of the Polysubstituted Benzene Ring Compound Represented by Formula II 19.89 g of crystalline form A of the polysubstituted benzene ring compound represented by Formula I was added to the reactor, followed by 137.4 g of anhydrous ethanol. The mixture was stirred and heated to 75°C. The internal temperature was maintained at 75°C. 4.0 g of maleic acid was added to the reactor via a feed hopper and stirred until a clear solution was obtained. The internal temperature of the reactor was lowered to 20°C, and the mixture was stirred until a large amount of solid precipitated. The internal temperature was maintained at 20°C, and 257.8 g of methyl t-butyl ether was added. After the dropwise addition was completed, the mixture was stirred for 0.5 to 3 hours. The raw material solution was placed in a press filter, and the mother liquor was filtered. The filter cake was washed with 60.1 g of methyl t-butyl ether, filtered, and dried to obtain 21.16 g of a solid sample.

[0098] Measurement of the X-ray powder diffraction spectrum revealed that the obtained sample was crystalline form B of the polysubstituted benzene ring compound maleate salt represented by formula II. The X-ray powder diffraction pattern shown at 2θ angles is shown in Figure 4.

[0099] Example 5: Preparation of crystalline form A of the maleate salt of the polysubstituted benzene ring compound represented by formula II Approximately 20 mg of crystalline form B of the maleate salt of the polysubstituted benzene ring compound represented by formula II obtained in Example 4 was weighed into an HPLC vial, 0.5 mL of isopropanol was added, and the resulting suspension was magnetically stirred (-1000 rpm) at 50°C for 6 days, and then centrifuged to obtain a sample.

[0100] The X-ray powder diffraction spectrum of the sample obtained by this method was compared with the spectrum of the sample in Example 2, and the sample was identified as crystalline form A of the polysubstituted benzene ring compound maleate salt represented by formula II.

[0101] Example 6: Preparation of crystalline form A of the polysubstituted benzene ring compound maleate salt represented by formula II Compared to Example 5, only the type of solvent was different. The isopropanol in Example 5 was replaced with isopropanol / water (a w =0.2), butanone, isopropyl acetate, anisole, acetonitrile, dichloromethane / cyclopentyl methyl ether (v:v=1:4), toluene, methyl isobutyl ketone, tetrahydrofuran, 1,4-dioxane, acetone, ethanol, methyl isobutyl ketone / isopropyl acetate (v:v=1:1), isopropanol / methyl t-butyl ether (v:v=1:1), or acetonitrile / toluene (v:v=1:1), respectively.

[0102] The X-ray powder diffraction spectrum of the sample obtained by this method was compared with the spectrum of the sample in Example 2, and both were identified as crystalline form A of the polysubstituted benzene ring compound maleate salt represented by formula II.

[0103] Example 7: Preparation of crystalline form A of the polysubstituted benzene ring compound maleate salt represented by formula II Compared to Example 5, only the temperature was different, and the 50°C in Example 5 was replaced with a temperature cycle, that is, 50°C to 5°C, 0.1°C / min, 3 cycles.

[0104] The X-ray powder diffraction spectrum of the sample obtained by this method was compared with the spectrum of the sample in Example 2, and both were identified as crystalline form A of the polysubstituted benzene ring compound maleate salt represented by formula II.

[0105] Example 8: Preparation of crystalline form A of the polysubstituted benzene ring compound maleate salt represented by formula II Compared with Example 5, only the temperature and reaction time were different, 50°C in Example 5 was replaced by room temperature, and the reaction time of 6 days was replaced by 30 days.

[0106] The X-ray powder diffraction spectrum of the sample obtained by this method was compared with the spectrum of the sample in Example 2, and both were identified as crystalline form A of the polysubstituted benzene ring compound maleate salt represented by formula II.

[0107] Example 9: Preparation of crystalline form A of the maleate salt of the polysubstituted benzene ring compound represented by formula II 20 mg of crystalline form B of the polysubstituted benzene ring compound maleate salt represented by Formula II obtained in Example 4 was weighed into a 3 mL vial, and 0.5 mL of a mixed solution of dimethyl sulfoxide and ethyl acetate (v:v = 1:3) was added. The mixture was stirred at 50°C for approximately 2 hours, and then filtered (using a PTFE filter membrane with a pore size of 0.45 μm) to obtain the filtrate. The obtained filtrate was placed in a biochemical incubator and cooled from 50°C to 5°C at a rate of 0.1°C / min. The sample was then transferred to room temperature and volatilized. A solid precipitated and was filtered to obtain the sample.

[0108] The X-ray powder diffraction spectrum of the sample obtained by this method was compared with the spectrum of the sample in Example 2, and the sample was identified as crystalline form A of the polysubstituted benzene ring compound maleate salt represented by formula II.

[0109] Example 10: Preparation of crystalline form A of the maleate salt of the polysubstituted benzene ring compound represented by formula II Crystalline Form B of the maleate salt of a polysubstituted benzene ring compound represented by Formula II obtained in Example 4 was weighed and mixed with a stock solution near saturation with a good solvent listed in Table 8. 1 mL of the stock solution was added to a 20 mL vial, and the corresponding antisolvent listed in Table 8 was added under magnetic stirring. The mixture was stirred dropwise until a solid precipitated. An "*" in Table 8 indicates that no solid was obtained even after approximately 9 mL of antisolvent was added, and the mixture was then transferred to 5°C to obtain a solid. Specific good and antisolvents are listed in Table 8.

[0110] [Table 8]

[0111] The X-ray powder diffraction spectrum of the sample obtained by this method was compared with the spectrum of the sample in Example 2, and both were identified as crystalline form A of the polysubstituted benzene ring compound maleate salt represented by formula II.

[0112] Example 11: Preparation of crystalline form A of the maleate salt of the polysubstituted benzene ring compound represented by formula II Crystalline Form B of the maleate salt of the polysubstituted benzene ring compound represented by Formula II obtained in Example 4 was weighed and mixed with a stock solution nearly saturated with a good solvent, methanol. 0.5 mL of the stock solution was added to a 3 mL vial, and approximately 4 mL of the antisolvent, acetone, was added to a 20 mL vial. The 3 mL vial was left open and placed in the 20 mL vial, which was then sealed and allowed to stand at room temperature. When solid precipitation was observed, the solid was collected and subjected to XRPD testing.

[0113] The X-ray powder diffraction spectrum of the sample obtained by this method was compared with the spectrum of the sample in Example 2, and the sample was identified as crystalline form A of the polysubstituted benzene ring compound maleate salt represented by formula II.

[0114] Example 12: Preparation of crystalline form A of the maleate salt of the polysubstituted benzene ring compound represented by formula II Compared with Example 11, only the type of anti-solvent was different, and the anti-solvent acetone in Example 11 was replaced with ethyl acetate or tetrahydrofuran, respectively.

[0115] The X-ray powder diffraction spectrum of the sample obtained by this method was compared with the spectrum of the sample in Example 2, and the sample was identified as crystalline form A of the polysubstituted benzene ring compound maleate salt represented by formula II.

[0116] Comparative Example 1: Compared with Example 5, only the type of solvent was different. The isopropanol in Example 5 was replaced with the solvents shown in Table 9, where "*" indicates that the mixture became clear after stirring and was evaporated to a solid when cooled to room temperature.

[0117] [Table 9]

[0118] Measurement of the X-ray powder diffraction spectrum revealed that the samples obtained with numbers 1 to 4 were all crystalline form C of the polysubstituted benzene ring compound maleate salt represented by formula II. The X-ray powder diffraction patterns were shown in FIG.

[0119] Comparative Example 2: Compared with Example 7, only the type of solvent was different, and the solvent in Example 7 was replaced with water.

[0120] The X-ray powder diffraction spectrum of the sample obtained by this method was compared with the spectrum of the sample in Comparative Example 1, and both were determined to be crystalline form C of the polysubstituted benzene ring compound maleate salt represented by formula II.

[0121] Comparative Example 3: Compared with Example 8, only the type of solvent was different. The solvent in Example 8 was replaced with the solvent shown in the table below, where "*" indicates that the solvent evaporated at room temperature to give a solid. The results are shown in Table 10.

[0122] [Table 10]

[0123] Measurement of the X-ray powder diffraction spectrum revealed that the sample obtained in No. 5 is crystalline form B of the poly-substituted benzene ring compound maleate represented by formula II. The X-ray powder diffraction spectra of the samples obtained in Nos. 1 to 4 were compared with the spectrum of the sample in Comparative Example 1, and all were determined to be crystalline form C of the poly-substituted benzene ring compound maleate represented by formula II.

[0124] Comparative Example 4: Compared with Example 9, only the type of solvent was different. The mixed solution of dimethyl sulfoxide and ethyl acetate (v:v=1:3) used as the solvent in Example 9 was replaced with the solvents shown in Table 11, where "*" indicates that a solid was directly precipitated after cooling to 5°C, and the results are shown in Table 11.

[0125] [Table 11]

[0126] The X-ray powder diffraction spectra of the samples obtained in numbers 1 to 4 were compared with the spectrum of the sample in Comparative Example 1, and all were identified as crystalline form C of the polysubstituted benzene ring compound maleate salt represented by formula II.

[0127] Comparative Example 5: Compared with Example 10, only the type of solvent was different. The good solvent and anti-solvent in Example 10 were replaced with the solvents shown in Table 12, respectively. Here, ** indicates that the mixture became clear when transferred to 5°C, and then transferred to room temperature for evaporation to obtain a solid. # indicates that the mixture became clear at 5°C, and due to the high boiling point of the solvent, was transferred to 50°C and vacuum dried to obtain a solid. The specific results are shown in Table 12.

[0128] [Table 12]

[0129] The X-ray powder diffraction spectra of the samples obtained in Nos. 1 to 11 were compared with the spectrum of the sample in Comparative Example 1, and the samples were determined to be crystalline form C of the poly-substituted benzene ring compound maleate represented by formula II. The X-ray powder diffraction spectrum of the sample obtained in No. 12 was compared with the spectrum of the sample in Comparative Example 3, and the sample was determined to be crystalline form B of the poly-substituted benzene ring compound maleate represented by formula II.

[0130] Comparative Example 6: The only difference from Example 11 was the type of solvent. The good solvent and anti-solvent in Example 11 were replaced with the solvents shown in Table 13, respectively. Here, * indicates that due to the high boiling point of the solvent, the mixture was transferred to 50°C and vacuum dried to obtain a solid. The results are shown in Table 13.

[0131] [Table 13]

[0132] The X-ray powder diffraction spectra of the samples obtained in numbers 1 to 6 were compared with the spectrum of the sample in Comparative Example 1, and all were identified as crystalline form C of the polysubstituted benzene ring compound maleate salt represented by formula II.

[0133] Comparative Example 7: Crystalline Form B of the maleate salt of the polysubstituted benzene ring compound represented by Formula II obtained in Example 4 was weighed into a 3 mL vial. Approximately 4 mL of the solvent shown in the table below was added to a 20 mL vial. The 3 mL vial was left open and placed in the 20 mL vial, and the 20 mL vial was then sealed. After standing at room temperature for 30 days, an XRPD test was performed, and the results are shown in Table 14.

[0134] [Table 14]

[0135] The X-ray powder diffraction spectrum of sample No. 1 was compared with the spectrum of sample No. 5 in Comparative Example 1, and all were determined to be crystalline form C of the poly-substituted benzene ring compound maleate represented by formula II. Measurement of the X-ray powder diffraction spectrum showed that sample No. 2 was crystalline form D of the poly-substituted benzene ring compound maleate represented by formula II. The X-ray powder diffraction patterns are shown in Figure 8. The X-ray powder diffraction spectra of samples Nos. 3 to 6 were compared with the spectrum of sample No. 5 in Comparative Example 3, and all were determined to be crystalline form B of the poly-substituted benzene ring compound maleate represented by formula II.

[0136] Comparative Example 8: Approximately 20 mg of crystalline form B of the maleate salt of the polysubstituted benzene ring compound represented by Formula II obtained in Example 4 was weighed into a 3 mL vial. Approximately 4 mL of saturated salt solution (40-90% RH) was added to a 20 mL vial. The 3 mL vial was left open and placed in the 20 mL vial, which was then sealed. After standing at room temperature for 30 days, an XRPD test was performed. The results are shown in Table 15.

[0137] [Table 15]

[0138] The X-ray powder diffraction spectra of the samples obtained in Nos. 1 and 2 were compared with the spectrum of sample No. 5 in Comparative Example 3, and all of them were determined to be crystalline form B of the poly-substituted benzene ring compound maleate represented by formula II. The X-ray powder diffraction spectra of the samples obtained in Nos. 3 and 4 were compared with the spectrum of sample No. 1 in Comparative Example 1, and all of them were determined to be crystalline form C of the poly-substituted benzene ring compound maleate represented by formula II.

[0139] Comparative Example 9: 20 mg of crystalline form B of the maleate salt of the polysubstituted benzene ring compound represented by Formula II obtained in Example 3 was weighed into a 3 mL glass vial, dissolved in 1.0 to 2.0 mL of each of the solvents listed in the table below, and filtered (through a PTFE membrane with a pore size of 0.45 μm). The filtrate was collected. The vial containing the clear solution was sealed with a sealing membrane, several small holes were drilled in the sealing membrane, and the solution was allowed to slowly evaporate at room temperature. When a solid precipitated, the resulting solid was collected and subjected to XRPD testing. The results are shown in Table 16.

[0140] [Table 16]

[0141] The X-ray powder diffraction spectrum of sample No. 1 obtained was compared with the spectrum of sample No. 5 in Comparative Example 3, and all of them were determined to be crystalline form B of the poly-substituted benzene ring compound maleate represented by formula II. The X-ray powder diffraction spectra of samples Nos. 2 to 5 obtained were compared with the spectrum of sample No. 1 in Comparative Example 1, and all of them were determined to be crystalline form C of the poly-substituted benzene ring compound maleate represented by formula II. The X-ray powder diffraction spectrum of samples Nos. 6 to 8 obtained were compared with the spectrum of sample No. 2 in Comparative Example 7, and all of them were determined to be crystalline form D of the poly-substituted benzene ring compound maleate represented by formula II.

[0142] Comparative Example 10: Crystalline form A of the polysubstituted benzene ring compound maleate salt represented by formula II obtained in Example 2 was heated to 187°C under N protection, and the sample obtained by X-ray powder diffraction spectrum was found to be crystalline form E of the polysubstituted benzene ring compound maleate salt represented by formula II. The X-ray powder diffraction pattern is shown in Figure 9.

[0143] Comparative Example 11: Preparation of Crystalline Form A of the Succinate Salt of the Polysubstituted Benzene Ring Compound Represented by Formula III

[0144] [ka]

[0145] 395.8 mg of the crystalline form A of the polysubstituted benzene ring compound represented by formula I obtained in Example 1 was weighed into a 20 mL glass vial, 88.0 mg of succinic acid was added, and 7.0 mL of ethyl acetate was added to form a suspension. The suspension was magnetically stirred at room temperature for about 3 days, and the solid was separated by suction filtration. The solid was dried under vacuum at room temperature overnight to obtain 418.9 mg of solid. X-ray powder diffraction spectrum measurement showed that the obtained sample was crystalline form A of the polysubstituted benzene ring compound succinate salt represented by formula III, and the X-ray powder diffraction pattern is shown in Figure 10. 1 H NMR data is 1 H NMR (400 MHz, DMSO-d6): δ 11.51 (s, 1H), 8.10 (t, J = 5.2Hz, 1H), 6.65 (d, J = 2.4Hz, 1H), 6.44 (d, J =2.4 Hz, 1H), 5.89 (s, 1H), 4.89-4.84 (m, 1H), 4.27 (d, J = 5.2Hz,2H), 3.85 (d, J =10.4Hz,2H), 3.33 (t, J = 10.8Hz, 2H), 3.01 (dd, J = 6.8, 13.6Hz,2H), 2.93-2.89 (m, 1H), 2.85-2.76 (m, 1H),2.42(s, 4H) 2.42-2.35 (m, 4H), 2.22 (s, 3H), 2.13 (d, J = 4.0, 6H), 1.63-1.46 (m, 10H), 0.81 (t, J = 7.2 Hz, 3H) ppm.

[0146] Comparative Example 12: Preparation of Crystalline Form A of the Hydrobromide Salt of the Polysubstituted Benzene Ring Compound Represented by Formula IV

[0147] [ka]

[0148] 400.7 mg of the crystalline form A of the polysubstituted benzene ring compound represented by formula I obtained in Example 1 was weighed into a 20 mL glass vial, 3.5 mL of ethyl acetate was added and mixed, 156.7 mg of hydrobromide was weighed, 3.5 mL of ethyl acetate was added and mixed, and the two portions of ethyl acetate were mixed to form a suspension, which was then magnetically stirred at room temperature for about 3 days and centrifuged to obtain a solid, which was then vacuum dried at room temperature overnight to obtain 318.6 mg of a solid. X-ray powder diffraction spectrum measurement confirmed that the obtained sample was crystalline form A of the polysubstituted benzene ring compound hydrobromide represented by formula IV, and the X-ray powder diffraction pattern is shown in Figure 11. 1 H NMR data is 1 H NMR (400 MHz, DMSO-d6): δ 11.51 (s, 1H), 8.10 (t, J = 5.2Hz, 1H), 6.65 (d, J = 2.4Hz, 1H), 6.44 (d, J =2.4 Hz, 1H), 5.89 (s, 1H), 4.89-4.84 (m, 1H), 4.27 (d, J = 5.2Hz,2H), 3.85 (d, J =10.4Hz,2H), 3.33 (t, J = 10.8Hz, 2H), 3.01 (dd, J = 6.8, 13.6Hz,2H), 2.93-2.89 (m, 1H), 2.85-2.76 (m, 4H), 2.42-2.35 (m, 2H), 2.22 (s, 3H), 2.13 (d, J = 4.0, 6H), 1.87-1.46 (m, 10H), 0.81 (t, J = 7.2Hz, 3H) ppm.

[0149] Comparative Example 13: Preparation of free crystalline form D of the polysubstituted benzene ring compound of formula I 400.7 mg of crystalline form A of the polysubstituted benzene ring compound represented by formula I obtained in Example 1 was weighed out and stirred in 7 mL of isopropyl acetate at room temperature for 5 days, and then separated by suction filtration to obtain a solid. The solid was dried under vacuum at room temperature overnight to obtain 330.0 mg of a solid. Measurement of the X-ray powder diffraction spectrum confirmed that the obtained sample was free crystalline form D represented by formula I. The X-ray powder diffraction pattern is shown in Figure 12. 1 H NMR data is1 H NMR (400 MHz, DMSO-d6): δ 11.48 (s, 1H), 8.05 (t, J = 5.2Hz, 1H), 6.59 (d, J = 2.4Hz, 1H), 6.38 (d, J =2.4 Hz, 1H), 5.68 (s, 1H), 4.68-4.68 (m, 1H), 4.24 (d, J = 5.2Hz,2H), 3.82 (d, J =10.4Hz,2H), 3.23 (t, J = 10.8Hz, 2H), 2.98 (dd, J = 6.8, 13.6Hz,2H), 2.93-2.92 (m, 1H), 2.91-2.90 (m, 1H), 2.29-2.26 (m, 8H), 2.19 (s, 3H), 2.11 (s, 3H), 2.10 (s, 3H), 1.61-1.39 (m, 10H), 0.79 (t, J = 7.2 Hz, 3H) ppm.

[0150] Comparative Example 14: Screening of free-state polycrystalline forms of polysubstituted benzene ring compounds of formula I Compared with Comparative Example 12, only the solvent and temperature were different, and the isopropyl acetate and room temperature in Comparative Example were replaced with the solvent and temperature in Table 17, respectively, and the results obtained are shown in Table 17.

[0151] [Table 17]

[0152] Measurement of X-ray powder diffraction spectra revealed that the obtained samples were free crystalline forms A, B, C, E, F, G, and I of the polysubstituted benzene ring compound represented by formula I, respectively, and the X-ray powder diffraction patterns of the obtained samples are shown in Figures 5 to 6, Figures 13 to 16, and Figure 18, respectively.

[0153] Comparative Example 15: Screening of Free-State Polycrystalline Forms of Polysubstituted Benzene Ring Compounds of Formula I The only difference compared to Comparative Example 13 No. 5 is that free crystalline form G of the poly-substituted benzene ring compound of Formula I obtained in Example 13 No. 5 can be heated to 160°C to obtain free crystalline form H of the poly-substituted benzene ring compound of Formula I. The X-ray powder diffraction pattern of the obtained sample is shown in Figure 17.

[0154] Effect Example 1: Screening of polycrystalline forms of the polysubstituted benzene ring compound maleate salt represented by formula II As in Examples 5 to 12 and Comparative Examples 1 to 10, five crystalline forms were discovered during the screening and crystalline form identification process, and were named crystalline forms A, B, C, D and E of the maleate salt, respectively, where A, B and E are anhydrous crystalline forms, and C and D are hydrate crystalline forms, where crystalline form E of the maleate salt can only be obtained at high temperature and can be reconverted to crystalline form A of the maleate salt upon cooling to room temperature.

[0155] Furthermore, to explore the interconversion relationship between each crystalline form and screen for a more stable crystalline form, suspension competition experiments were conducted on the anhydrous crystalline forms A and B of maleate, and suspension competition experiments were also conducted on the superior anhydrous crystalline form A of maleate and the hydrate crystalline forms C and D of maleate at different water activities.

[0156] Suspension competition experiment between anhydrous crystalline forms A and B of the maleate salt: Four 5 mL vials were taken, and 2 mL each of IPA and IPAc was added. Excess crystalline form B of the maleate salt was added, and the mixture was stirred at room temperature / 50°C for 24 hours, followed by filtration to obtain saturated solutions of the maleate salt in IPA and IPAc. Approximately 10 mg each of crystalline forms A and B of the maleate salt were added to each saturated solution, and the mixture was suspended and stirred at room temperature / 50°C for 3 days before being tested by XRPD. The results are shown in Table 18.

[0157] [Table 18]

[0158] As a result, all reaction systems yielded crystalline form A of the maleate, which was more stable than crystalline form B under room temperature / 50°C conditions.

[0159] Suspension competition experiment between anhydrous maleate crystalline form A and hydrate maleate crystalline forms C and D: Six 5 mL vials were taken and immersed in water at different water activities (a w To each of the saturated solutions, 2 mL of an ethanol solution of each of the maleate salts (0 / 0.2 / 0.4 / 0.6 / 0.8 / 1) was added, followed by excess maleate form B. The mixture was stirred at room temperature for 24 hours and then filtered to obtain saturated solutions. Approximately 10 mg of maleate form A, C, or D was added to each saturated solution, and the mixture was suspended and stirred at room temperature for two days before being tested by XRPD. Two 5 mL vials were each filled with 2 mL of water, followed by excess maleate form B. The mixture was stirred at 50°C and 80°C for 20 hours and 2 hours, respectively, and then filtered to obtain saturated solutions. Approximately 10 mg of maleate form A, C, or D was added to each saturated solution, followed by suspension and stirring at 50°C and 80°C for 1 day and 3.5 hours, respectively, before being tested by XRPD. The results are shown in Table 19.

[0160] [Table 19]

[0161] As a result, crystalline form A of the maleate salt was obtained when the water activity was 0.2 or less, and crystalline form C of the maleate salt was obtained when the water activity was 0.4 or more in water at 50°C / 80°C. Both crystalline forms C and D of the maleate salt were converted to crystalline form A when suspended and stirred in an ethanol solution with a water activity of 0.2 or in absolute ethanol at room temperature.

[0162] From the above examples and suspension competition experiments, it was found that among the five crystalline forms of maleate, crystalline form E can only be obtained at high temperatures and reconverts to crystalline form A upon cooling to room temperature; crystalline forms A and B have similar solubility, but crystalline form A has better solid properties, being a granular solid (crystalline form B is a powdery solid), and crystalline form A is more stable than crystalline form B; crystalline form B can be converted to crystalline form A in multiple solvents and under multiple conditions; crystalline forms C and D are easily converted to crystalline form A; and crystalline form C can be converted to crystalline form A after drying the hydrate at 170°C. Therefore, among the five crystalline forms of maleate, crystalline form A of maleate is the most stable.

[0163] Effect Example 2: Physicochemical stability Appropriate amounts of crystalline Form A of the polysubstituted benzene ring compound maleate salt of Formula II were weighed and left open at 60°C for one day, then at 25°C / 60%RH and 40°C / 75%RH for one week. After storage under different conditions, the solid samples were tested for purity by HPLC to evaluate chemical stability, and the crystalline form by XRPD to evaluate physical stability. The evaluation results are summarized in Table 20.

[0164] [Table 20]

[0165] As a result, under the three test conditions, the crystalline form A of the polysubstituted benzene ring compound maleate salt represented by formula II did not undergo any crystalline form transformation or decrease in chemical purity, indicating that it has high physical and chemical stability.

[0166] Effect Example 3: Moisture absorption Through salt form screening experiments, multiple salt forms of the polysubstituted benzene ring compound represented by formula I were obtained. For different salt forms, crystallinity, TGA weight loss, DSC thermal signal, and salt formation molar ratio were comprehensively considered, and crystalline form A of the polysubstituted benzene ring compound maleate represented by formula II, crystalline form A of the polysubstituted benzene ring compound succinate represented by formula III, and crystalline form A of the polysubstituted benzene ring compound hydrobromide represented by formula IV were screened, and the following effect experiments were carried out:

[0167] Approximately 10 mg of each of the crystalline form A of the polysubstituted benzene ring compound maleate salt represented by formula II, the crystalline form A of the polysubstituted benzene ring compound succinate salt represented by formula III, the crystalline form A of the polysubstituted benzene ring compound hydrobromide salt represented by formula IV, and the free crystalline form D of the polysubstituted benzene ring compound represented by formula I was taken and dried at a temperature of 25°C and a humidity of 0%RH. The moisture absorption characteristics of the sample were tested when the humidity was changed from 0%RH to 95%RH, and when the humidity was changed from 0%RH to 95%RH and back to 0%RH. If the mass change rate over time was less than 0.002%, the sample was deemed to be in balance when the humidity change step size was 10%RH and the mass change rate dm / dt was less than 0.002%. If the mass change rate within 10 minutes was less than 0.002% / min, it was deemed to be in balance. The balance criterion for the detection process was a mass change rate less than 0.002% / min. The maximum balance time was 3 hours. The water isothermal adsorption / desorption characteristics under the test conditions were measured, and the samples after DVS test were detected by XRPD. The results are shown in Table 21.

[0168] [Table 21]

[0169] As a result, among the four crystalline forms, crystalline form A of the maleate salt had the lowest hygroscopicity, at 25°C / 80%

[0170] The water adsorption of RH was 0.4%, which means that it has almost no hygroscopicity, and the crystal form did not change after the DVS test.

[0171] Effect Example 4: Dynamic Solubility Four samples, namely, Form A of the polysubstituted benzene ring compound maleate salt represented by Formula II, Form A of the polysubstituted benzene ring compound succinate salt represented by Formula III, Form A of the polysubstituted benzene ring compound hydrobromide salt represented by Formula IV, and Form D of the polysubstituted benzene ring compound free crystals represented by Formula I, were tested for dynamic solubility in three biosolvents (SGF, FaSSIF, and FeSSIF) and water at 37°C. For each test, 45 mg of solid and 4.5 mL of solvent were mixed in a 5 mL centrifuge tube. The centrifuge tube was sealed and fixed on a rotating disk rotating at 40 rpm, and the rotating disk was placed in a 37°C thermostatic bath. Samples were taken at four time points: 1, 2, 4, and 24 hours after equilibration. At each sampling point, the filtrate was separated and tested for concentration and pH, and the crystalline form was tested for the solid.

[0172] As a result, in water, crystalline form A of the succinate salt is clear (>8.44 mg / mL), the solubilities of crystalline form A of the maleate salt and crystalline form A of the hydrobromide salt are similar (6.99 mg / mL and 4.26 mg / mL), and the solubility of free crystalline form D is low (0.016 mg / mL).

[0173] In SGF, all three salt forms were clear (>7.83 mg / mL), with free crystalline form D having a relatively low solubility (6.98 mg / mL).

[0174] In FaSSIF, the succinate and maleate forms A have high solubilities (5.39 mg / mL and 4.15 mg / mL), while the hydrobromide form A and free form D have low solubilities (1.39 mg / mL and 0.42 mg / mL).

[0175] In FeSSIF, Form A of the succinate, Form A of the maleate, and Form D in the free state were clear (>7.35 mg / mL), while Form A of the hydrobromide had a relatively low solubility (5.33 mg / mL).

[0176] To summarize the above results, the solubility of maleate crystalline form A and succinate crystalline form A in the four media was relatively high, reaching 6 mg / mL in water and SGF, which met the dissolution sink condition even when the dosage specification was not more than 600 mg, allowing for complete in vitro release. The solubility reached 4 mg / mL in FaSSIF and FeSSIF. However, a comparison of the overall hygroscopicity revealed that succinate crystalline form A had high hygroscopicity, which was unfavorable for subsequent formulation process development. However, maleate crystalline form A had low hygroscopicity, and when the D90 was controlled at 200 μm, the API had good fluidity and the angle of repose of a single API was around 40°. Therefore, the formulation process for both forms could be carried out using dry blending and direct pressure, making them more suitable for the development of herbal formulation processes.

Claims

1. Crystalline form A of the maleate salt of a polysubstituted benzene ring compound represented by formula II, wherein the X-ray powder diffraction pattern shown in 2θ angles has diffraction peaks at 7.39±0.2°, 8.78±0.2°, 13.99±0.2°, 15.21±0.2°, 15.73±0.2°, 17.75±0.2°, 18.51±0.2°, and 21.06±0.2°. Crystalline form A of a polysubstituted benzene ring compound maleate salt represented by formula II, characterized in that: 【Chemistry 1】

2. an X-ray powder diffraction pattern shown in 2θ angles also having diffraction peaks at one or more 2θ angles of 8.51±0.2° and 16.06±0.2°; and / or the crystalline form A of the polysubstituted benzene ring compound maleate salt represented by formula II has major endothermic peaks at 185.3°C ± 3°C and 204.4°C ± 3°C in a differential scanning calorimetry diagram.

2. The crystalline form A of the polysubstituted benzene ring compound maleate salt represented by formula II according to claim 1.

3. The X-ray powder diffraction pattern shown in 2θ angles also has diffraction peaks at one or more 2θ angles of 18.20±0.2° and 20.68±0.2°.

2. The crystalline form A of the polysubstituted benzene ring compound maleate salt represented by formula II according to claim 1.

4. The X-ray powder diffraction pattern shown in terms of 2θ angles also has diffraction peaks at one or more 2θ angles of 22.81±0.2° and 25.28±0.2°.

2. The crystalline form A of the polysubstituted benzene ring compound maleate salt represented by formula II according to claim 1.

5. The X-ray powder diffraction patterns shown at 2θ angles are shown in Table 1-1 and Table 1-2 below. The crystalline form A of the polysubstituted benzene ring compound maleate salt represented by formula II according to any one of claims 1 to 4. Table 1-1 Table 1-2

6. A method for preparing crystalline form A of the polysubstituted benzene ring compound maleate salt of formula II according to any one of claims 1 to 5, which is the following method 1, method 2, method 3 or method 4: Method 1: A method of reacting a polysubstituted benzene ring compound represented by formula I with maleic acid in solvent A, 【Chemistry 2】 the method, wherein the solvent A is one selected from methyl isobutyl ketone, 2-methyltetrahydrofuran, and ethyl acetate, and the reaction is carried out at room temperature; Method 2: A method comprising suspending crystalline form B of the maleate salt of the poly-substituted benzene ring compound of formula II in solvent B and stirring the suspension, wherein the solvent B is one selected from isopropanol, isopropanol / water, butanone, isopropyl acetate, anisole, acetonitrile, dichloromethane / cyclopentyl methyl ether, toluene, methyl isobutyl ketone, tetrahydrofuran, 1,4-dioxane, acetone, ethanol, methyl isobutyl ketone / isopropyl acetate, isopropanol / methyl t-butyl ether, and acetonitrile / toluene, wherein the isopropanol / water has an a w of 0.2, the volume ratio of dichloromethane to cyclopentyl methyl ether in the dichloromethane / cyclopentyl methyl ether is 1:4, the volume ratio of methyl isobutyl ketone to isopropyl acetate in the methyl isobutyl ketone / isopropyl acetate is 1:1, the volume ratio of isopropanol to methyl t-butyl ether in the isopropanol / methyl t-butyl ether is 1:1, and the volume ratio of acetonitrile to toluene in the acetonitrile / toluene is 1:1, the method, wherein the stirring is carried out at room temperature, 50°C, or a temperature cycle of 50°C to 5°C at 0.1°C / min, wherein when the stirring temperature is 50°C, the stirring time is 5 to 8 days, and when the stirring temperature is room temperature, the stirring time is 25 to 35 days, and when the stirring is carried out at a temperature cycle, the temperature cycle conditions are 50°C to 5°C, 0.1°C / min, and 2 to 4 cycles; Method 3: A method of crystallizing Form B of the polysubstituted benzene ring compound maleate salt of Formula II by saturating it in a mixed solvent of dimethyl sulfoxide and ethyl acetate, wherein the crystallization comprises slowly lowering the temperature from 50°C to 5°C at a rate of 0.1°C / min, and if the saturated solution is still clear, transferring it to room temperature and volatilizing it; Method 4: A method for dissolving crystalline form B of the poly-substituted benzene ring compound maleate salt of formula II in solvent C, adding solvent D, and crystallizing the resulting mixture, wherein the solvent C is one selected from methanol, dimethyl sulfoxide, and trichloromethane; when the solvent C is methanol, the solvent D is ethyl acetate; when the solvent C is dimethyl sulfoxide, the solvent D is isopropyl acetate; when the solvent C is trichloromethane, the solvent D is one or more of ethyl acetate, acetonitrile, and butanone; After adding solvent D, a solid precipitates, and if no solid precipitates, the temperature is lowered to 5°C, and if no solid precipitates, the mixture is evaporated at room temperature or dried under vacuum at 50°C. The crystalline form B of the polysubstituted benzene ring compound maleate salt represented by formula II is a crystal having the following X-ray powder diffraction pattern: Table 2 。

7. In method 1, the reaction time is 2 to 4 days, and / or in Method 1, the molar ratio of the poly-substituted benzene ring compound represented by Formula I to the maleic acid is 1:(1 to 1.2); and / or in Method 1, the volume / mass ratio of the solvent A to the poly-substituted benzene ring compound represented by Formula I is 15 to 40 mL / g; and / or in method 2, the stirring is magnetic stirring; and / or in Method 2, the volume / mass ratio of the solvent B to the crystalline form B of the poly-substituted benzene ring compound maleate salt represented by formula II is 20 to 50 mL / g; and / or in the method 3, the volume ratio of dimethyl sulfoxide to ethyl acetate in the mixed solvent is 1:(2-5); and / or in Method 3, the volume / mass ratio of the mixed solvent to the crystalline form B of the poly-substituted benzene ring compound maleate salt represented by Formula II is 20 to 50 mL / g; and / or in Method 4, the solvent D is added dropwise; and / or in the method 4, the volume ratio of the solvent D to the solvent C is (7 to 15):1; and / or wherein the methods 1 to 4 include a post-treatment step of separation by centrifugation or filtration and drying by vacuum drying; 7. A method for preparing crystalline form A of the polysubstituted benzene ring compound maleate salt of formula II according to claim 6.

8. In method 1, the molar ratio of the polysubstituted benzene ring compound represented by formula I to the maleic acid is 1:1.07; and / or in Method 1, the volume / mass ratio of the solvent A to the poly-substituted benzene ring compound represented by Formula I is 17.6 mL / g; and / or in Method 2, the volume / mass ratio of the solvent B to the crystalline form B of the poly-substituted benzene ring compound maleate salt represented by formula II is 25 mL / g; and / or in the method 3, the volume ratio of dimethyl sulfoxide to ethyl acetate in the mixed solvent is 1:3; and / or in Method 3, the volume / mass ratio of the mixed solvent to the crystalline form B of the poly-substituted benzene ring compound maleate salt represented by Formula II is 25 mL / g; and / or, in Method 4, the volume ratio of Solvent D to Solvent C is 9:1; 8. A method for preparing crystalline form A of the polysubstituted benzene ring compound maleate salt of formula II according to claim 7.

9. Method 2, 3, or 4 further comprises the steps of dissolving a poly-substituted benzene ring compound represented by formula I and maleic acid in ethanol, lowering the temperature to cause crystallization, and adding methyl t-butyl ether to prepare crystalline form B of the poly-substituted benzene ring compound maleate represented by formula II.

7. A method for preparing crystalline form A of the polysubstituted benzene ring compound maleate salt of formula II according to claim 6.

10. In preparing the crystalline form B of the poly-substituted benzene ring compound maleate salt represented by formula II, the molar ratio of the poly-substituted benzene ring compound represented by formula I to the maleic acid is 1:(0.95-0.98); and / or, in preparing the crystalline form B of the poly-substituted benzene ring compound maleate salt represented by formula II, the mass ratio of the ethanol to the poly-substituted benzene ring compound represented by formula I is (7-9):1; and / or, in the preparation of the crystalline form B of the poly-substituted benzene ring compound maleate salt represented by formula II, the temperature of the temperature-lowering crystallization is 15 to 25°C; and / or, in preparing the crystalline form B of the poly-substituted benzene ring compound maleate salt represented by formula II, the mass ratio of the methyl t-butyl ether to the poly-substituted benzene ring compound represented by formula I is (12-15):1; and / or the preparation of crystalline form B of the poly-substituted benzene ring compound maleate salt represented by formula II includes post-treatment steps of filtration and drying.

10. The method for preparing crystalline form A of the polysubstituted benzene ring compound maleate salt of formula II according to claim 9.

11. In preparing the crystalline form B of the polysubstituted benzene ring compound maleate salt represented by formula II, the molar ratio of the polysubstituted benzene ring compound represented by formula I to maleic acid is 1:0.978; and / or, in preparing the crystalline form B of the poly-substituted benzene ring compound maleate salt represented by formula II, the mass ratio of the ethanol to the poly-substituted benzene ring compound represented by formula I is 6.9:1; and / or, in the preparation of the crystalline form B of the poly-substituted benzene ring compound maleate salt represented by formula II, the temperature of the temperature-lowering crystallization is 20°C; and / or, in preparing the crystalline form B of the poly-substituted benzene ring compound maleate salt represented by formula II, the mass ratio of the methyl t-butyl ether to the poly-substituted benzene ring compound represented by formula I is 13:1; and / or the preparation of crystalline form B of the poly-substituted benzene ring compound maleate salt represented by formula II includes a post-treatment step of further adding methyl t-butyl ether to the filter cake obtained after filtration to wash it.

11. A method for preparing crystalline form A of the polysubstituted benzene ring compound maleate salt of formula II according to claim 10.

12. A pharmaceutical composition comprising crystalline form A of the polysubstituted benzene ring compound maleate salt represented by formula II according to any one of claims 1 to 5 and a pharmaceutically acceptable carrier. A pharmaceutical composition comprising:

13. A pharmaceutical composition comprising crystalline form A of the polysubstituted benzene ring compound maleate salt of formula II according to any one of claims 1 to 5 for preventing or treating an EZH2-mediated disease.

14. The EZH2-mediated disease includes cancer. The pharmaceutical composition of claim 13.

15. The cancers include brain cancer, thyroid cancer, cardiac sarcoma, lung cancer, oral cancer, stomach cancer, liver cancer, kidney cancer, pancreatic cancer, esophageal cancer, nasopharyngeal cancer, laryngeal cancer, colorectal cancer, breast cancer, prostate cancer, bladder cancer, ovarian cancer, uterine cancer, bone cancer, melanoma, glioblastoma, lymphoma, blood cancer, adrenal neuroblastoma, skin cancer, astrocytoma; 15. The pharmaceutical composition of claim 14.

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