Sulfate crystal form of an amide compound, pharmaceutical composition and use thereof

Abstract

The present invention belongs to the field of pharmaceutical chemistry, and relates to different sulfate crystal forms of an amide compound, namely, 1-(3-cyano-5-methylthiophene-2-yl)-N-(6-methoxy-1H-benzo[d]imidazol-2-yl)-2,5-dimethyl-1H-pyrrole-3-carboxamide, a pharmaceutical composition containing the crystal form, and uses thereof. The chemical stability and crystal stability of the sulfate crystal form are both better than those of the free base crystal form.

Description

Technical Field

[0001] The present invention belongs to the field of pharmaceutical chemistry and relates to different sulfate salt forms of an amide compound, namely, 1-(3-cyano-5-methylthiophene-2-yl)-N-(6-methoxy-1H-benzo[d]imidazol-2-yl)-2,5-dimethyl-1H-pyrrole-3-carboxamide (hereinafter referred to as "the compound of formula (1)"), and also relates to a pharmaceutical composition containing the crystal form and its medical use. Background Art

[0002] DHX33 belongs to the RNA helicase protein family containing the DEAD / H box. DEAD / H stands for the amino acid abbreviation Asp-Glu-Ala-Asp / His. This sequence, together with several other conserved amino acid sequences, appears in the protein sequences of RNA helicase family members and is highly involved in nucleic acid substrate binding and ATP hydrolysis. Although these family members share these same sequences, each RNA helicase has its own specific specificity and unique biological function. The molecular weight of human DHX33 protein is 72kDa. It has the function of unwinding nucleic acids. It uses the bioenergy released by ATP hydrolysis to drive the change of the conformation of RNA and protein complexes, and then participates in a variety of RNA metabolic activities, specifically, a series of biological processes from RNA transcription, shearing, editing, translation to degradation. The function of DHX33 is not limited to the modification of RNA molecules. Studies have shown that in addition to unwinding RNA double strands, DHX33 protein is also involved in DNA metabolism. Specifically, DHX33 protein can unwind the double-stranded structure of DNA and play an important role in gene expression.

[0003] Studies have shown that DHX33 affects the methylation state of DNA by binding to the promoters of genes related to various cancers, thereby regulating the expression of various cancer genes and signal pathways related to tumor development at the genomic level, and plays a vital role in various cell activities such as cell growth, proliferation, migration, apoptosis, and metabolism. In addition, it was found that DHX33 can sense the invasion of foreign double-stranded RNA molecules and play an important role in the innate immunity of cells. As a very important cell growth regulatory gene, DHX33 is highly expressed in many cancers, such as lung cancer, lymphoma, glioblastoma, breast cancer, colon cancer, liver cancer, etc. The occurrence and development of many cancers depend on the high expression of DHX33 protein. Genetic knockout of DHX33 can significantly inhibit the occurrence and development of lung cancer driven by RAS oncogene; in vivo and in vitro experiments have confirmed that after inhibiting DHX33 protein, the occurrence and development of various cancers such as breast cancer, colon cancer, brain glioma, lymphoma, etc. are significantly inhibited. Since the protein function of DHX33 depends on its helicase activity, and the helicase activity-deficient mutant of DHX33 does not have the function of DHX33 protein, it cannot replace the function of the protein encoded by the wild-type DHX33 gene.

[0004] The applicant has discovered a variety of compounds that can inhibit the RNA helicase activity of DHX33 (for example, 1-(3-cyano-5-methylthiophene-2-yl)-N-(6-methoxy-1H-benzo[d]imidazol-2-yl)-2,5-dimethyl-1H-pyrrole-3-carboxamide), and verified that these compounds can significantly inhibit the growth and proliferation of cancer cells both in vitro and in vivo. Summary of the invention

[0005] The object of the present invention is to provide different sulfate salt forms of an amide compound, namely 1-(3-cyano-5-methylthiophene-2-yl)-N-(6-methoxy-1H-benzo[d]imidazol-2-yl)-2,5-dimethyl-1H-pyrrole-3-carboxamide (hereinafter referred to as "the compound of formula (1)"), a pharmaceutical composition containing the same and uses thereof.

[0006] The structural formula of the compound of formula (1) is as follows:

[0007]

[0008] In a first aspect, the present invention provides a sulfate crystalline form A of the compound of formula (1), whose X-ray powder diffraction (XRPD) pattern has characteristic diffraction peaks at the following 2θ angles: 5.5±0.2° and 11.0±0.2°.

[0009] In some embodiments, the X-ray powder diffraction pattern of the sulfate crystalline form A of the compound of formula (1) further has characteristic diffraction peaks at one or more of the following 2θ angles: 21.5±0.2°, 22.0±0.2° and 23.6±0.2°.

[0010] In some embodiments, the X-ray powder diffraction pattern of the sulfate crystalline form A of the compound of formula (1) further has characteristic diffraction peaks at one or more of the following 2θ angles: 10.2±0.2°, 13.0±0.2° and 18.8±0.2°.

[0011] The present invention also provides a method for preparing the sulfate crystal form A of the compound of formula (1), comprising: adding an organic solvent to the free crystal form A of the compound of formula (1), then adding sulfuric acid, stirring at room temperature for about 2-5 days, collecting the solid and then drying to obtain the sulfate crystal form A. In a specific embodiment of the present invention, the organic solvent is selected from one or both of acetone and isopropanol.

[0012] In a second aspect, the present invention provides a sulfate salt form B of the compound of formula (1), whose X-ray powder diffraction (XRPD) pattern has characteristic diffraction peaks at the following 2θ angles: 5.5±0.2°, 16.1±0.2° and 21.0±0.2°.

[0013] In some embodiments, the X-ray powder diffraction pattern of the sulfate crystalline form B of the compound of formula (1) further has characteristic diffraction peaks at one or more of the following 2θ angles: 11.8±0.2°, 25.2±0.2° and 26.8±0.2°.

[0014] In some embodiments, the X-ray powder diffraction pattern of the mesylate salt form B of the compound of formula (1) also has characteristic diffraction peaks at one or more of the following 2θ angles: 15.0±0.2°, 19.2±0.2° and 23.8±0.2°.

[0015] The present invention also provides a method for preparing the sulfate crystal form B of the compound of formula (1), comprising: adding an organic solvent to the free crystal form A of the compound of formula (1), then adding sulfuric acid, stirring at room temperature for about 2-5 days, collecting the solid and then drying to obtain the sulfate crystal form B. In a specific embodiment of the present invention, the organic solvent is acetonitrile.

[0016] In an embodiment of the present invention, in the above preparation method, stirring may be performed at room temperature for about 1-4 days, for example, about 1, 1.5, 2, 2.5, 3, 3.5, 4 days.

[0017] In an embodiment of the present invention, in the above preparation method, drying can be carried out under vacuum at 40-95° C. for about 8-16 hours. In a specific embodiment, drying can be carried out under vacuum at 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95° C. for about 8, 8.5, 9, 9.5, 10, 10.5, 11, 11.5, 12, 12.5, 13, 13.5, 14, 14.5, 15, 15.5, 16 hours.

[0018] In a third aspect, the present invention provides a pharmaceutical composition comprising at least one of the above-mentioned crystal forms and one or more pharmaceutically acceptable carriers. The composition can be used to treat and / or prevent diseases or conditions at least partially mediated by DHX33, such as cancer, viral infection and inflammation, such as breast cancer, colon cancer, brain glioma, lymphoma, etc.

[0019] In a fourth aspect, the present invention provides use of the above-mentioned crystal form or the above-mentioned pharmaceutical composition in the preparation of a medicament for preventing and / or treating a disease or condition at least partially mediated by DHX33.

[0020] In a fifth aspect, the present invention provides a method for preventing and / or treating a disease or condition at least partially mediated by DHX33, comprising the following steps: administering a preventive and / or therapeutically effective amount of the above-mentioned crystalline form or the above-mentioned pharmaceutical composition to an individual in need thereof; preferably, the disease is selected from cancer, viral infection and inflammation mediated by DHX33.

[0021] The present invention is not to be limited to the particular embodiments described herein; it is also to be understood that the terminology used herein is for the purpose of describing and not limiting the particular embodiments.

[0022] Definition of terms

[0023] Unless otherwise stated, the technical and scientific terms used herein have the same meaning as those skilled in the art to which the present invention belongs. If there is a contradiction, the definition provided by the application shall prevail. When a certain amount, concentration or other value or parameter is expressed in the form of a range, a preferred range or a preferred upper numerical limit and a preferred lower numerical limit, it should be understood that it is equivalent to specifically disclose any scope by combining any pair of upper range limits or preferred values ​​with any range lower limit or preferred values, without considering whether the scope is specifically disclosed. Unless otherwise stated, the numerical range listed herein is intended to include all integers and fractions (decimals) within the endpoints and the range of the range.

[0024] The term "about" when used with a numerical variable generally refers to the value of that variable and all values ​​of that variable are within experimental error (eg, within a 95% confidence interval for the mean) or within ±10% of the specified value, or wider.

[0025] The expression "comprising" or its synonymous similar expressions "including", "containing" and "having" etc. are open-ended and do not exclude additional unrecited elements, steps or ingredients. The expression "consisting of excludes any element, step or ingredient not specified. The expression "consisting essentially of means that the scope is limited to the specified elements, steps or ingredients, plus the optional elements, steps or ingredients that do not materially affect the basic and novel characteristics of the claimed subject matter. It should be understood that the expression "comprising" encompasses the expressions "consisting essentially of" and "consisting of".

[0026] The term "X-ray powder diffraction pattern (XRPD pattern)" refers to an experimentally observed diffraction pattern or a parameter, data or value derived therefrom. An XRPD pattern is typically characterized by peak positions (abscissa) and / or peak intensities (ordinate).

[0027] The term "diffraction angle" or "2θ" refers to the peak position expressed in degrees (°) based on the setup in the X-ray diffraction experiment, and is usually the unit of the abscissa in the diffraction pattern. If the reflection is diffracted when the incident beam forms an angle θ with a certain lattice plane, the experimental setup requires that the reflected beam be recorded at an angle of 2θ. It should be understood that the specific 2θ value mentioned in this article for a specific crystal form is intended to represent the 2θ value (expressed in degrees) measured using the X-ray diffraction experimental conditions described herein.

[0028] It should be noted that in the powder X-ray diffraction spectrum, the position of the peak or the relative intensity of the peak may vary due to factors such as the measuring instrument, measuring method / conditions, etc. For any specific crystal form, there may be an error in the position of the peak, and the measurement error of the 2θ value may be ±0.2°. Therefore, when determining each crystal form, this error should be taken into account, and the error is also within the scope of this application.

[0029] For the same crystal form, the position of the endothermic peak of DSC may be different due to factors such as the measuring instrument, measuring method / conditions, etc. For any specific crystal form, the position of the endothermic peak may have an error, which may be ±5°C or ±3°C. Therefore, when determining each crystal form, this error should be taken into account, and the error is also within the scope of this application.

[0030] For the same crystal form, the position of the weight loss temperature of TGA may be different due to factors such as the measuring instrument, measuring method / conditions, etc. For any specific crystal form, there may be an error in the position of the weight loss temperature, which may be ±5°C or ±3°C. Therefore, when determining each crystal form, this error should be taken into account, and the error is also within the scope of this application.

[0031] It should be understood that different types of equipment or using different test conditions may give slightly different XRPD patterns and characteristic peaks or different DSC patterns and characteristic peaks. The specific values ​​provided are not to be taken as absolute values.

[0032] The term "room temperature" refers to 20°C ± 5°C.

[0033] The term "prevention" refers to prophylactic administration to reduce the likelihood of or delay the onset of a disease or symptom.

[0034] The term "treatment" aims to alleviate or eliminate the disease state or condition being targeted. It should also be understood that the treatment of the disease state or condition includes not only complete treatment, but also less than complete treatment, but achieving some biological or medically relevant results.

[0035] The term "pharmaceutically acceptable" refers to those compounds, materials, compositions and / or crystal forms that are suitable for use in contact with human and animal tissues without excessive toxicity, irritation, allergic reaction or other problems or complications, and are commensurate with a reasonable benefit / risk ratio, within the scope of sound medical judgment. "Pharmaceutically acceptable carrier" refers to an inert substance that is administered together with the active ingredient and is conducive to the administration of the active ingredient, including but not limited to any glidant, sweetener, diluent, preservative, dye / colorant, flavor enhancer, surfactant, wetting agent, dispersant, disintegrant, suspending agent, stabilizer, isotonic agent, solvent or emulsifier that is acceptable for human or animal (e.g., livestock) approved by the State Food and Drug Administration.

[0036] The above-mentioned pharmaceutical compositions can act systemically and / or locally. For this purpose, they can be administered by suitable routes, for example, by parenteral, topical, intravenous, oral, subcutaneous, intraarterial, intradermal, percutaneous, rectal, intracranial, intraperitoneal, intranasal, intramuscular routes or as inhalant administration.

[0037] The above-mentioned administration route can be achieved through a suitable dosage form. The dosage forms that can be used in the present invention include, but are not limited to, tablets, capsules, lozenges, hard candies, powders, sprays, creams, ointments, suppositories, gels, pastes, lotions, ointments, aqueous suspensions, injectable solutions, elixirs, syrups, etc.

[0038] When administered orally, the pharmaceutical composition can be prepared into any orally acceptable preparation form, including but not limited to tablets, capsules, aqueous solutions, aqueous suspensions, and the like.

[0039] The term "disease or condition mediated at least in part by DHX33" refers to a disease whose pathogenesis at least partially involves factors related to DHX33, such as cancer, viral infection, and inflammation.

[0040] The term "effective amount" refers to a dose that can induce a biological or medical response in cells, tissues, organs or organisms (eg, individuals) and is sufficient to achieve the desired preventive and / or therapeutic effect.

[0041] The therapeutically effective amount of the crystalline form described in the present application is from about 0.0001-20 mg / Kg body weight / day, for example, from 0.001-10 mg / Kg body weight / day.

[0042] The dosage frequency of the crystal formation described in the present application can be determined by a physician according to the needs of individual patients, for example, once or twice a day, or more times a day. Administration can be intermittent, for example, wherein during a period of several days, the patient receives a daily dose of the crystal formation, and then during a period of several days or more days, the patient does not receive a daily dose of the crystal formation. BRIEF DESCRIPTION OF THE DRAWINGS

[0043] Figure 1 It is the X-ray powder diffraction (XRPD) pattern of the free crystalline form A of the compound of formula (1).

[0044] Figure 2 This is a thermogravimetric analysis (TGA) diagram of the free crystalline form A of the compound of formula (1).

[0045] Figure 3 It is a differential scanning calorimetry (DSC) diagram of the free crystalline form A of the compound of formula (1).

[0046] Figure 4 It is the free form A of the compound of formula (1) 1 HNMR spectrum.

[0047] Figure 5 It is the X-ray powder diffraction (XRPD) pattern of the sulfate crystal form A of the compound of formula (1).

[0048] Figure 6 This is a thermogravimetric analysis (TGA) diagram of the sulfate crystal form A of the compound of formula (1).

[0049] Figure 7 It is a differential scanning calorimetry (DSC) diagram of sulfate crystal form A of the compound of formula (1).

[0050] Figure 8 It is the sulfate crystal form A of the compound of formula (1) 1 HNMR spectrum.

[0051] Fig. 9 It is the X-ray powder diffraction (XRPD) pattern of the sulfate crystal form B of the compound of formula (1).

[0052] Fig.10This is a thermogravimetric analysis (TGA) diagram of the sulfate crystal form B of the compound of formula (1).

[0053] Fig.11 It is a differential scanning calorimetry (DSC) diagram of the sulfate crystal form B of the compound of formula (1).

[0054] Fig.12 It is the sulfate crystal form B of the compound of formula (1) 1 HNMR spectrum.

[0055] Fig.13 It is a comparison diagram of XRPD of the solid before and after the equilibrium solubility test of the sulfate crystal form A of the compound of formula (1).

[0056] Fig.14 This is a XRPD comparison chart of the stability of the sulfate crystal form A of the compound of formula (1) before and after storage.

[0057] Fig.15 This is the DVS test diagram of the sulfate crystal form A of the compound of formula (1).

[0058] Fig.16 It is a comparison diagram of XRPD of sulfate crystal form A of the compound of formula (1) before and after DVS test.

[0059] Fig.17 This is a comparison of the XRPD diagrams of the stability of the free base crystal form A of the compound of formula (1) before and after storage. DETAILED DESCRIPTION

[0060] The intermediate compounds of the present application can be prepared by a variety of synthesis methods well known to those skilled in the art, including the specific embodiments listed below, embodiments formed by combining them with other chemical synthesis methods, and equivalent substitution methods well known to those skilled in the art. Preferred embodiments include but are not limited to the examples of the present application.

[0061] The chemical reactions of the specific embodiments of the present application are carried out in a suitable solvent, which must be suitable for the chemical changes of the present application and the reagents and materials required. In order to obtain the compounds of the present application, it is sometimes necessary for those skilled in the art to modify or select the synthesis steps or reaction processes based on the existing embodiments.

[0062] The present application will be described in detail below through examples, which do not imply any limitation to the present application.

[0063] X-ray powder diffraction (XRPD)

[0064] XRPD Testing Parameters

[0065]

[0066]

[0067] Thermogravimetric analysis (TGA) and differential scanning calorimetry (DSC) TGA and DSC test parameters

[0068]

[0069] H NMR spectrum ( 1 HNMR)

[0070] 1 HNMR test parameters

[0071]

[0072] Ion Chromatography (IC)

[0073] IC Test Parameters

[0074]

[0075]

[0076] High Performance Liquid Chromatography (HPLC)

[0077] HPLC test parameters

[0078]

[0079] Dynamic Water Sorption Analysis (DVS)

[0080] DVS Test Parameters

[0081]

[0082] Example 1: Free Form A of the Compound of Formula (I) (1-(3-cyano-5-methylthiophene-2-yl)-N-(6-methoxy-1H-benzo[d]imidazol-2-yl)-2,5-dimethyl-1H-pyrrole-3-carboxamide) and Preparation Method

[0083] (1) Preparation method of compound 3 (ethyl 2-acetyl-4-pentanoate)

[0084]

[0085] Compound 1 (ethyl acetoacetate) (5 g, 38.42 mmol, 1.0 eq) was dissolved in triethylamine (75 mL), and compound 2 (chloroacetone) (3.5 g, 38.42 mmol, 1.0 eq) was added. The reactants were reacted at 110 ° C for 2 hours under nitrogen protection. After concentration, the residue was dissolved in water (100 mL) and then extracted twice with dichloromethane (50 mL each time). The organic layer was washed with brine, dried over anhydrous sodium sulfate, filtered and then concentrated. The residue was purified by flash column chromatography (petroleum ether / ethyl acetate = 100 / 1 to 50 / 1 to 20 / 1) to obtain compound 3 (ethyl 2-acetyl-4-pentanoate) (1.3 g, yield: 18.3%) as a colorless oil. MS (ESI) m / z: 187 [M+H + ]TLC: PE / EA(2 / 1); R f (Compound 1) = 0.6; R f (Compound 3) = 0.4.

[0086] (2) Preparation method of compound 5 (1-(3-cyano-5-methylthiophene-2-yl)-2,5-dimethyl-1H-pyrrole-3-carboxylic acid ethyl ester)

[0087]

[0088] Compound 3 (2-acetyl-4-pentanoic acid ethyl ester) (1 g, 7.23 mmol, 1.0 eq) was dissolved in toluene (20 mL), and compound 4 (2-amino-3-cyano-5-methylthiophene) (1.6 g, 8.68 mmol, 1.2 eq) and p-toluenesulfonic acid (249 mg, 1.45 mmol, 0.2 eq) were added. The reactants were stirred at 110 ° C for 16 hours. The solid was filtered and concentrated. The residue was purified by flash column chromatography (petroleum ether / ethyl acetate = 50 / 1 to 30 / 1) to obtain yellow oily compound 5 (1-(3-cyano-5-methylthiophene-2-yl)-2,5-dimethyl-1H-pyrrole-3-carboxylic acid ethyl ester) (860 mg, yield: 41%). MS (ESI) m / z: 289 [M+H + ]. TLC: petroleum ether / ethyl acetate (10 / 1); R f (Compound 3) = 0.2; R f (Compound 5) = 0.4.

[0089] (3) Preparation method of free crystal form A of the compound of formula (I)

[0090]

[0091] Trimethylaluminum (0.15 mL, 0.306 mmol, 1.0 eq, 2 M in toluene) was added to compound 5 (ethyl 1-(3-cyano-5-methylthiophene-2-yl)-2,5-dimethyl-1H-pyrrole-3-carboxylate) (50 mg, 0.306 mmol, 1.0 eq) and compound 7 (5-methoxy-1H-benzimidazol-2-amine) (88 mg, 0.306 mmol, 1.0 eq) dissolved in 1 mL of toluene. The reactants were stirred at 100°C for 16 hours. The mixture was cooled to room temperature, quenched with methanol (10 mL), and then adjusted to pH 3 with 3 M hydrochloric acid. The mixture was diluted with water (30 mL) and then extracted three times with ethyl acetate, 20 ml each time. The organic layer was washed with brine, dried over anhydrous sodium sulfate, filtered and concentrated. The residue was purified by preparative high pressure liquid phase Prep-HPLC (acetonitrile / water (containing 0.1% formic acid)) to obtain free crystalline form A (5 mg, yield: 4%) of the compound of formula (I). MS (ESI) m / z: 406 [M+H + ]. 1 H NMR (400MHz, DMSO-d6): δ12.04(s,1H),11.20(s,1H),7.32(s,1H),7.29(s,1H),6.98(s,1H ), 6.89 (s, 1H), 6.69 (d, J = 7.2Hz, 1H), 3.73 (s, 3H), 2.47 (s, 3H), 2.38 (s, 3H), 2.04 (s, 3H).

[0092] The free crystalline form A has substantially the following Figure 1 XRPD pattern shown.

[0093] The XRPD pattern of free crystalline form A has characteristic diffraction peaks at the following 2θ angles, as shown in Table 1:

[0094] Table 1

[0095]

[0096] like Figure 2 As shown, the thermogravimetric analysis (TGA) curve of the free crystalline form A shows a weight loss of about 1.02% when the temperature is raised to about 200°C.

[0097] like Figure 3 As shown, the differential scanning calorimetry (DSC) curve of the free crystalline form A has an endothermic peak at about 163°C.

[0098] The free form A of the compound of formula (1) 1 HNMR images Figure 4 shown.

[0099] Example 2: Sulfate salt of the compound of formula (I) and its preparation method

[0100] Preparation method of sulfate crystal form A of compound of formula (I)

[0101] Weigh about 20 mg of free crystal form A into a 3 mL glass vial, add 1.2 equivalents (5.78 mg) of sulfuric acid, add 0.5 mL of acetone, ultrasonically promote sample dissolution, stir at room temperature for about 2 days, centrifuge to obtain a solid, and place it at 60 ° C for 12 hours in a vacuum dryer. Separate and test XRPD, which is sulfate crystal form A. This method replaces acetone with isopropanol, and sulfate crystal form A can also be prepared.

[0102] Sulfate Form A has substantially Figure 5 XRPD pattern shown.

[0103] The XRPD pattern of sulfate crystal form A has characteristic diffraction peaks at the following 2θ angles, as shown in Table 2-1:

[0104] Table 2-1

[0105]

[0106] like Figure 6 As shown, the thermogravimetric analysis (TGA) curve of sulfate crystal form A shows a weight loss of about 0.3390% when the temperature is raised to about 150° C. Sulfate crystal form A is an anhydrous substance.

[0107] like Figure 7 As shown, the differential scanning calorimetry (DSC) curve of sulfate salt form A has an endothermic peak at about 233.42°C.

[0108] Sulfate Form A 1 H-NMR spectrum Figure 8 As shown. Figure 8 It can be seen that the structure of this compound is no different from the free base compound.

[0109] Preparation method of sulfate crystal form B of compound of formula (I)

[0110] About 20 mg of free crystal form A was weighed into a 3 mL glass vial, 1.2 equivalents (5.78 mg) of sulfuric acid was added, 0.5 mL of acetonitrile was added, and the sample was dissolved by ultrasound. After stirring at room temperature for about 2 days, the solid was obtained by centrifugation and placed in a vacuum dryer at 60°C for 12 hours. The XRPD test showed that the sulfate crystal form B was obtained.

[0111] Sulfate Form B has substantially the following Fig. 9 The XRPD pattern of sulfate crystal form B has characteristic diffraction peaks at the following 2θ angles, as shown in Table 2-2.

[0112] Table 2-2

[0113]

[0114] like Fig.10 As shown, the thermogravimetric analysis (TGA) curve of sulfate crystal form B shows a weight loss of about 4.7791% when the temperature is raised to about 140° C. Sulfate crystal form B is a hydrate or anhydrous, but water is adsorbed on the surface.

[0115] like Fig.11 As shown, the differential scanning calorimetry (DSC) curve of sulfate salt form B has multiple endothermic peaks at about 31-114°C and melts at 205.98°C.

[0116] The molar ratio of the compound of formula (I) to sulfuric acid in the sulfate salt form B is about 1:1.

[0117] Sulfate Form B 1 H-NMR spectrum Fig.12 From the NMR, the crystal compound has no structural difference from the free base.

[0118] Example 3: 24-hour equilibrium solubility experiment in water

[0119] The 24-hour equilibrium solubility of free base crystal form A and sulfate crystal form A was tested. The specific steps are: weigh 10 mg of free base crystal form A and sulfate crystal form A respectively into 1 mL of water to prepare a suspension, stir magnetically at 37±2°C, sample after 24 hours, centrifuge the sample, and use the obtained solid for XRPD testing. The supernatant is filtered through a 0.22 μm PTFE filter membrane and then tested for solubility. The results are shown in Table 3. The solubility of free base crystal form A is about 1.92 μg / mL, and the solubility of sulfate crystal form A is 0.56 μg / mL.

[0120] Table 3

[0121] Solid form Equilibrium solubility in water at 37℃ (μg / mL) Free base form A 1.92 Sulfate Form A 0.56

[0122] XRPD comparison of the solid before and after the equilibrium solubility test of sulfate crystal form A, such as Fig.13 shown.

[0123] Example 4: One-week stability experiment

[0124] Free base crystal form A and sulfate crystal form A samples were placed under 25°C / 60%RH (long term) and 40°C / 75%RH (accelerated) conditions for one week, and then tested for HPLC purity and crystal form changes. The results are shown in Table 4. After sulfate crystal form A was placed under the two test conditions for one week, there was no significant change in HPLC purity and crystal form. The chemical stability and crystal form stability of sulfate crystal form A were both good, and the chemical stability was better than that of free base. The XRPD comparison of the stability sample before and after placement is shown in Table 4. Fig.14 shown.

[0125] Table 4

[0126]

[0127] Example 5: Hygroscopicity test

[0128] To evaluate the stability risk of samples at 25°C with humidity changes, a DVS test was performed on sulfate crystal form A, and solid samples after the test were collected for XRPD testing. The results are shown in Table 5.

[0129] Under 0-30% RH conditions, the water absorption of sulfate crystal form A is less than 0.5%, and the sample is slightly hygroscopic.

[0130] The sulfate crystal form A absorbs 1.5-2% water under 80-90% RH conditions.

[0131] After DVS testing, the crystal form of the sulfate form A sample did not change significantly (the difference in XRPD was mainly caused by the preferred orientation and crystallinity).

[0132] DVS and XRPD test results are as follows Figure 15-16 The XRPD comparison of the stability of the free base crystal form A of the compound of formula (1) before and after storage is shown in FIG. Fig.17 shown.

[0133] Table 5

[0134]

Claims

1. A sulfate crystal form A of an amide compound, wherein the structural formula of the amide compound is as shown in formula (1): Its X-ray powder diffraction pattern has characteristic diffraction peaks at the following 2θ angles: 5.5±0.2° and 11.0±0.2°, The X-ray powder diffraction pattern also has characteristic diffraction peaks at one or more of the following 2θ angles: 21.5±0.2°, 22.0±0.2° and 23.6±0.2°, and The X-ray powder diffraction pattern also has characteristic diffraction peaks at one or more of the following 2θ angles: 10.2±0.2°, 13.0±0.2° and 18.8±0.2°.

2. The crystalline form A according to claim 1, wherein the sulfate crystalline form A has the XRPD pattern shown in Figure 5.

3. A pharmaceutical composition comprising the crystalline form A according to claim 1 and a pharmaceutically acceptable carrier.

4. Use of the crystalline form A according to claim 1 or the pharmaceutical composition according to claim 3 in the preparation of a medicament for preventing and / or treating lung cancer, lymphoma, glioblastoma, breast cancer, colon cancer, or liver cancer.

5. The method for preparing the sulfate crystal form A according to claim 1, comprising: An organic solvent is added to the free crystalline form A of the compound of formula (1), and then sulfuric acid is added, and the mixture is stirred at room temperature for 2-5 days. The solid is collected and then dried to obtain the sulfate crystalline form A, wherein the organic solvent is selected from acetone or isopropanol.

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