A crystalline form of a mesylate salt of a polycyclic compound, pharmaceutical compositions, and uses thereof

Abstract

The application belongs to the field of pharmaceutical chemistry, and relates to different methanesulfonic acid salt crystal forms of a polycyclic compound (E)-2-(3-(2-cyano-2-(6-methoxy-3H-imidazo[4,5-c]pyridin-2-yl)vinyl)-2,5-dimethyl-1H-pyrrol-1-yl)-5-methylthiophene-3-carbonitrile, pharmaceutical compositions containing the crystal forms and purposes. The purity, water solubility, chemical stability and crystal form stability of the methanesulfonic acid salt crystal form are all better than those of other crystal forms.

Description

TECHNICAL FIELD

[0001] The present application belongs to the field of pharmaceutical chemistry, and relates to different mesylate salt crystal forms of a polycyclic compound, namely (E)-2-(3-(2-cyano-2-(6-methoxy-3H-imidazo[4,5-c]pyridin-2-yl)vinyl)-2,5-dimethyl-1H-pyrrol-1-yl)-5-methylthiophene-3-carbonitrile (hereinafter referred to as "compound of formula (I)"), as well as to pharmaceutical compositions comprising the crystal forms and medical uses. BACKGROUND

[0002] DHX33 belongs to the DEAD / H box-containing RNA helicase protein family. Among them, DEAD / H represents the abbreviation of amino acids Asp-Glu-Ala-Asp / His, which appears in the protein sequence of the members of the RNA helicase family, 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 72 kDa, which has the function of unwinding nucleic acids, and uses the biological energy released by ATP hydrolysis to drive the conformational change of RNA and protein complexes, and is involved in various RNA metabolic activities, specifically from RNA transcription, splicing, editing, translation to degradation, etc. 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 plays an important role in gene expression.

[0003] Research has shown that DHX33 binds to the promoters of various cancer-related genes, affecting DNA methylation. This in turn regulates the expression of numerous cancer genes and signaling pathways associated with tumor development at the genomic level, playing a crucial role in numerous cellular activities, including cell growth, proliferation, migration, apoptosis, and metabolism. Furthermore, DHX33 has been found to sense the intrusion of foreign double-stranded RNA molecules and play a crucial role in cellular innate immunity. As a crucial cell growth regulator, DHX33 is highly expressed in a variety of cancers, including lung cancer, lymphoma, glioblastoma, breast cancer, colon cancer, and liver cancer. The development and progression of many cancers depend on elevated DHX33 protein expression. Genetic knockout of DHX33 significantly inhibits the development and progression of RAS oncogene-driven lung cancer. In vitro and in vivo experiments have confirmed that inhibition of DHX33 significantly suppresses the development and progression of various cancers, including breast cancer, colon cancer, glioma, and lymphoma. 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, (E)-2-(3-(2-cyano-2-(6-methoxy-3H-imidazo[4,5-c]pyridin-2-yl)vinyl)-2,5-dimethyl-1H-pyrrol-1-yl)-5-methylthiophene-3-carbonitrile), 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 methanesulfonate crystalline forms of a polycyclic compound, namely (E)-2-(3-(2-cyano-2-(6-methoxy-3H-imidazo[4,5-c]pyridin-2-yl)vinyl)-2,5-dimethyl-1H-pyrrol-1-yl)-5-methylthiophene-3-carbonitrile (hereinafter referred to as "compound of formula (I)"), a pharmaceutical composition containing the crystalline form, and uses.

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

[0007]

[0008] In a first aspect, the present invention provides a mesylate salt crystalline form 1 of the compound of formula (I), whose X-ray powder diffraction (XRPD) pattern has characteristic diffraction peaks at the following 2θ angles: 4.6±0.2° and 18.3±0.2°.

[0009] In some embodiments, the X-ray powder diffraction pattern of the methanesulfonic acid salt Form 1 of the compound of Formula (I) further comprises one or more characteristic diffraction peaks at two-theta angles of 21.2 ± 0.2°, 25.9 ± 0.2°, and 28.0 ± 0.2°.

[0010] In some embodiments, the X-ray powder diffraction pattern of the methanesulfonic acid salt Form 1 of the compound of Formula (I) further comprises one or more characteristic diffraction peaks at two-theta angles of 4.1 ± 0.2°, 19.8 ± 0.2°, 24.1 ± 0.2°, and 26.3 ± 0.2°.

[0011] The present application also provides a method for preparing the methanesulfonic acid salt Form 1 of the compound of Formula (I), comprising: adding an organic solvent to the free form of the compound of Formula (I), then adding methanesulfonic acid, stirring at room temperature for about 2-5 days, collecting the solid and drying to obtain the methanesulfonic acid salt Form 1. In a specific embodiment of the present application, the organic solvent is isopropanol.

[0012] In an embodiment of the present application, in the above preparation method, the stirring can be carried out at room temperature for about 1-4 days, for example, about 1, 1.5, 2, 2.5, 3, 3.5, 4 days.

[0013] In an embodiment of the present application, in the above preparation method, the drying can be carried out at 40-95°C under vacuum for about 8-16 hours. In a specific embodiment, the drying can be carried out at 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95°C under vacuum 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.

[0014] In a third aspect, the present application 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 for treating and / or preventing a disease or disorder mediated at least in part by DHX33, such as cancer, viral infection and inflammation, and the cancer can be, for example, breast cancer, colon cancer, glioma, lymphoma, etc.

[0015] In a fourth aspect, the present application 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 disorder mediated at least in part by DHX33.

[0016] In a fifth aspect, the present application provides a method for preventing and / or treating a disease or disorder mediated at least in part by DHX33, comprising the step of: administering a prophylactically and / or therapeutically effective amount of the above-mentioned crystal form or the above-mentioned pharmaceutical composition to an individual in need thereof; preferably, the disease is selected from the group consisting of cancer, viral infection and inflammation mediated by DHX33.

[0017] The application is not limited to the particular embodiments described herein; it should also be understood that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of particular embodiments.

[0018] Definitions of terms

[0019] Unless defined otherwise, technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application belongs. If there is a conflict between the definitions provided in this application and those provided in the art to which this application pertains, the definitions provided in this application control. When expressing a quantity, concentration, or other value or parameter of interest in the form of a range, a preferred range, or a preferred upper limit and a preferred lower limit, it is to be understood that every range of values, preferred or otherwise, between the stated lower value and the stated upper value, is intended to be specifically recited. Unless otherwise stated, numerical ranges expressed in the form "from x to y" are understood to include x and y. Numerical ranges expressed in the form "x or more" or "y or less" are understood to include x and y, respectively.

[0020] The term "about" when used in connection with a numerical value, generally refers to the numerical value and all numerical values within experimental error (e.g., within 95% confidence interval of the mean) or within ±10% of the indicated numerical value, or within a broader range.

[0021] The recitation of "comprising" or analogous terms such as "comprises", "comprised of", "including", "including", "containing", "containing", "having" and "having" are open-ended, and do not exclude additional, unrecited elements, steps, or ingredients. The recitation "consisting of excludes any element, step, or ingredient not specified. The recitation "consisting essentially of limits the scope of the specified elements, steps, or ingredients, plus optional elements, steps, or ingredients that do not materially affect the basic and novel characteristics of the claimed subject matter. It is to be understood that the recitation "comprising" encompasses the recitations "consisting essentially of" and "consisting of".

[0022] The term "X-ray powder diffraction pattern (XRPD pattern)" refers to an experimentally observed diffraction pattern or parameters, data, or values derived therefrom. An XRPD pattern is typically characterized by peak positions (abscissa) and / or peak intensities (ordinate).

[0023] The term "diffraction angle" or "2Θ" refers to the peak position in degrees (°) based on the experimental setup in an X-ray diffraction experiment, and is typically the unit of abscissa in a diffraction pattern. The experimental setup requires recording the reflected beam at a 2Θ angle if the reflection is diffracted when the incident beam forms a Θ angle with a certain lattice plane. It is to be understood that a particular 2Θ value mentioned herein for a particular crystalline form is intended to mean the 2Θ value (in degrees) measured using the X-ray diffraction experimental conditions described herein.

[0024] It should be noted that the position of the peaks or the relative intensity of the peaks in the powder X-ray diffraction spectrum can vary due to the measuring instrument, measuring method / conditions, and the like. For any particular crystal form, the position of the peaks can have an error, and the error in the measurement of the 2Θ value can be ±0.2°. Therefore, in determining each crystal form, this error should be taken into account, and within the error, it also falls within the scope of the present application.

[0025] For the same crystal form, the position of the endothermic peak in the DSC can vary due to the measuring instrument, measuring method / conditions, and the like. For any particular crystal form, the position of the endothermic peak can have an error, and the error can be ±5°C, and can be ±3°C. Therefore, in determining each crystal form, this error should be taken into account, and within the error, it also falls within the scope of the present application.

[0026] For the same crystal form, the position of the weight loss temperature in the TGA can vary due to the measuring instrument, measuring method / conditions, and the like. For any particular crystal form, the position of the weight loss temperature can have an error, and the error can be ±5°C, and can be ±3°C. Therefore, in determining each crystal form, this error should be taken into account, and within the error, it also falls within the scope of the present application.

[0027] It should be understood that different types of equipment or with different test conditions can give slightly different XRPD patterns and characteristic peaks or different DSC patterns and characteristic peaks. The specific values provided cannot be taken as absolute values.

[0028] The term "room temperature" refers to 20°C ± 5°C.

[0029] The term "prevention" refers to prophylactic administration to reduce the likelihood of the onset of a disease or symptoms, or to delay the onset of the disease or symptoms.

[0030] The term "treatment" is intended to refer to the alleviation or eradication of the disease state or condition being addressed. It is also to be appreciated that the treatment of a disease state or condition includes a partial but significant therapeutic effect.

[0031] The term "pharmaceutically acceptable" refers to those compounds, materials, compositions and / or crystalline forms that are suitable for use in contact with human and animal tissues, without excessive toxicity, irritation, allergic reactions 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 facilitates 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 use in humans or animals (e.g., livestock) as approved by the State Food and Drug Administration.

[0032] Above-mentioned pharmaceutical composition can act systemically and / or act topically.For this purpose, they can be administered by suitable approach, for example by parenteral, local, intravenous, oral, subcutaneous, intraarterial, intradermal, percutaneous, rectal, intracranial, intraperitoneal, intranasal, intramuscular approach or as inhalant administration.

[0033] The above-mentioned route of administration can be achieved through suitable dosage forms. 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.

[0034] 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.

[0035] 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.

[0036] 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.

[0037] The therapeutically effective amount of the crystalline form described herein is from about 0.0001-20 mg / Kg body weight / day, for example, from 0.001-10 mg / Kg body weight / day.

[0038] The dosage frequency of the crystalline forms described herein can be determined by a physician based on the needs of the individual patient, for example, once or twice a day, or more times a day. Administration can be intermittent, for example, wherein the patient receives a daily dose of the crystalline form over a period of several days, followed by a period of several or more days in which the patient does not receive a daily dose of the crystalline form. BRIEF DESCRIPTION OF THE DRAWINGS

[0039] Figure 1 is an X-ray powder diffraction (XRPD) pattern of Form 1 sulfate salt of the compound of Formula (I).

[0040] Figure 2 is an X-ray powder diffraction (XRPD) pattern of Form 1 methanesulfonate salt of the compound of Formula (I). 1 HNMR pattern.

[0041] Figure 3 is an X-ray powder diffraction (XRPD) pattern of Form 1 sulfate salt of the compound of Formula (I).

[0042] Figure 4 is an X-ray powder diffraction (XRPD) pattern of Form 2 sulfate salt of the compound of Formula (I).

[0043] Figure 5 is an X-ray powder diffraction (XRPD) pattern of Form 4 sulfate salt of the compound of Formula (I).

[0044] Figure 6 is a thermogravimetric analysis (TGA) pattern of Form 1 methanesulfonate salt of the compound of Formula (I). 1 HNMR pattern.

[0045] Figure 7 is an X-ray powder diffraction (XRPD) pattern of Form 1 methanesulfonate salt of the compound of Formula (I).

[0046] Figure 8 is an X-ray powder diffraction (XRPD) pattern of Form 2 methanesulfonate salt of the compound of Formula (I).

[0047] Figure 9 is a thermogravimetric analysis (TGA) pattern of Form 1 methanesulfonate salt of the compound of Formula (I).

[0048] Figure 10 is a differential scanning calorimetry (DSC) pattern of Form 1 methanesulfonate salt of the compound of Formula (I).

[0049] Figure 11 is a crystalline form comparison of Form 1 methanesulfonate salt of the compound of Formula (I) in stability analysis. 1 H NMR pattern.

[0050] Figure 12 is a crystalline form comparison of Form 1 methanesulfonate salt of the compound of Formula (I) in stability analysis.

[0051] Figure 13 is a hygroscopic DVS test pattern of Form 1 methanesulfonate salt of the compound of Formula (I).

[0052] Figure 14 is an XRPD comparison of Form 1 methanesulfonate salt of the compound of Formula (I) before and after DVS testing. DETAILED DESCRIPTION

[0053] The intermediate compounds of the present application can be prepared by a variety of synthetic methods well known to those skilled in the art, including the specific embodiments set forth below, embodiments formed by a combination of the specific embodiments with other chemical synthetic methods well known to those skilled in the art, and equivalents thereof as appreciated by those skilled in the art, preferred embodiments including but not limited to the examples of the present application.

[0054] The chemical reactions of the specific embodiments of the present application are performed in solvents appropriate to the reagents and materials employed and suitable for the chemical changes being effected. In the event that solvents are not specifically named, the skilled artisan will be aware of the solvents that are suitable for the reactions being performed and the reagents and materials being used. In order to obtain the compounds of the present application, the synthetic procedures or reaction sequences of the embodiments can need to be modified or selected by those skilled in the art based on the embodiments provided.

[0055] The present application will now be described in detail by way of specific examples which are not meant to be limiting in any way.

[0056] X-ray powder diffraction (XRPD)

[0057] XRPD test parameters

[0058]

[0059] Thermogravimetric analysis (TGA) and differential scanning calorimetry (DSC)

[0060] TGA and DSC test parameters

[0061]

[0062] H NMR spectrum ( 1 HNMR) 1 H NMR test parameters

[0063]

[0064] Ion chromatography (IC)

[0065] IC test parameters

[0066]

[0067] High performance liquid chromatography (HPLC) HPLC test parameters

[0068]

[0069]

[0070] Dynamic vapour sorption analysis (DVS)

[0071] DVS test parameters

[0072]

[0073] Example 1: Synthesis of compound of formula (I) and its identification

[0074] 1.1 The synthesis route is as follows:

[0075]

[0076] wherein R is

[0077] 1.2 Synthesis method:

[0078] 1.2.1 Preparation of compound 2 (6-methoxypyridine-3,4-diamine)

[0079]

[0080] Compound 1 (2-methoxy-5-nitropyridin-4-amine) (3.0 g, 17.74 mmol, 1.0 eq.) was dissolved in methanol (30 mL), and carbon supported palladium catalyst (300 mg, 0.1 wt%) was added. The mixture was stirred at room temperature for 16 h in the presence of hydrogen. After filtration, the solid was concentrated to give compound 2 (6-methoxypyridine-3,4-diamine) (2.6 g, yield: 100%) as a brown solid. MS (ESI) m / z: 140 [M+H]+; TLC: DCM:MeOH (10:1); Rf(compound 1)=0.7; Rf(compound 2)=0.5. f f : compound 3) = 0.5.

[0081] 1.2.2 Preparation of compound 4 (2-(6-methoxy-3H-imidazo[4,5-c]pyridin-2- yl)acetonitrile)

[0082]

[0083] Compound 2 (6-methoxypyridine-3,4-diamine) (1.5 g, 10.2 mmol, 1.0 eq.) and compound 3 (ethyl cyanoacetate) (3.5 g, 30.6 mmol, 3.0 eq.) were dissolved in dimethylformamide (6 mL) and stirred at 180 °C for 5 h. After cooling, the solvent was removed. The residue was purified by flash column chromatography (dimethylformamide:methanol = 200:1 to 50:1) to give compound 4 (2-(6-methoxy-3H-imidazo[4,5-c]pyridin-2-yl)acetonitrile) (500 mg, yield: 26.0%) as a brown solid powder. MS (ESI) m / z: 188 [M+H]+; TLC: petroleum ether / ethyl acetate (1:1); Rf(compound 1)=0.5; Rf(compound 4)=0.5. f : compound 3) = 0.5.​f : (Compound 3) = 0.2.

[0084] 1.2.3, Synthesis of (E)-2-(3-(2-cyano-2-(6-methoxy-3H-imidazo[4,5-c]pyridin-2-yl)vinyl)-2,5-dimethyl-1H-pyrrol-1-yl)- methylthiophene-3-carbonitrile (Compound A)

[0085]

[0086] Compound 4 (2-(6-methoxy-3H-imidazo[4,5-c]pyridin-2-yl)acetonitrile) (80 mg, 0.42 mmol, 1.0 eq.) was dissolved in 1 mL of ethanol, Compound 5 (2-(3-formyl-2,5-dimethyl-1H-pyrrol-1-yl)- methylthiophene-3-carbonitrile) (103 mg, 0.42 mmol, 1.0 eq.) and piperidine (36 mg, 0.42 mmol, 1.0 eq.) were added. The mixture was heated to reflux and stirred for 1 h. After the reaction was completed, the reaction was cooled to room temperature and filtered. The solid was collected and dried to give Compound AB24288 ((E)-2-(3-(2-cyano-2-(6-methoxy-3H-imidazo[4,5-c]pyridin-2-yl)vinyl)-2,5-dimethyl-1H-pyrrol-1-yl)- methylthiophene-3-carbonitrile) (95 mg, yield: 13.6%) as a yellow powder.

[0087] 1 H NMR (400 MHz, DMSO-d6) δ 8.53 (s, 1H), 8.16 (s, 1H), 7.31 (s, 1H), 6.96 (s, 1H), 6.89 (s, 1H), 3.89 (s, 3H), 2.51 (s, 3H), 2.30 (s, 3H), 2.08 (s, 3H).

[0088] The free form of the compound of formula (I) is in amorphous state, having an XRPD pattern substantially as shown in Figure 1 .

[0089] The free form of the compound of formula (I) 1 H NMR pattern as shown in Figure 2 .

[0090] In view of the poor aqueous solubility of the free form of the weak base compound of formula (I), in order to improve the aqueous solubility of the compound and facilitate its bioavailability, two strong acids, sulfuric acid and methanesulfonic acid, were used to carry out salt formation reaction, aiming to improve the physicochemical properties of the compound and facilitate the later formulation development.

[0091] Example 2: Sulfate salt crystal form of the compound of formula (I) and its preparation method

[0092] Process for preparing a sulfate salt crystalline form of a compound of formula (I)

[0093] Pilot test: about 20 mg of free form compound of formula (I) and a certain molar amount of counterion (1.1 eq.) were weighed in a 3.0 mL vial, 1.0 mL of solvent (isopropyl alcohol, acetone and ethyl acetate were selected) was added and stirred magnetically at room temperature, after stirring for 2 days, the obtained solid was separated and dried at 50°C under blast, and then XRPD was measured. Three sulfate salt crystal forms 1 / 2 / 3 (weak crystal state) were obtained, among which crystal form 1 was obtained by using isopropyl alcohol as the solvent ( Figure 3 ); crystal form 2 was obtained by using acetone as the solvent ( Figure 4 ); and crystal form 3 was obtained by using ethyl acetate as the solvent, and since the crystallinity was very poor, no XRPD spectrum was provided. From the XRPD spectrum, the purity of the sulfate salt crystal form 1 was poor, so acetone was selected as the solvent for the later scale-up synthesis of the sulfate salt crystal form 2 of the compound of formula (I).

[0094] Scale-up synthesis: 199.92 mg of free form compound of formula (I) was weighed and added into 5 mL of acetone to form a suspension; 10 mL of acetone solution containing 29.0 μL of sulfuric acid (1.1 eq.) was added under stirring at room temperature; after 4 days of stirring under cyclic temperature rising and falling, the system was turbid, centrifuged, and dried at 50°C under blast for 2 hours; the solid was crushed and dried at 50°C under blast overnight. The solid was separated and tested by XRPD, and the crystal form 2 obtained in the pilot test was converted into sulfate salt crystal form 4.

[0095] The sulfate salt crystal form 4 has an XRPD pattern substantially as shown in Figure 5 .

[0096] The XRPD pattern of the sulfate salt crystal form 4 has characteristic diffraction peaks at the following 2θ angles, as shown in Table 1:

[0097] Table 1

[0098]

[0099] The H-NMR spectrum of the sulfate salt crystal form 4 is shown in 1 . It can be seen from Figure 6 that the structure of the compound is not different from that of the free base compound. Figure 6

[0100] The HPLC detection results show that the HPLC purity of the sulfate salt is significantly reduced during the drying process, and the solid state stability is poor. The HPLC purity data are shown in Table 2:

[0101] Table 2

[0102] Solid form HPLC purity Free base (amorphous) 96.20 Sulfate salt crystalline form 4 75.10 ​

[0103] Example 3: Preparation and characterization of the methanesulfonic acid salt Form 1 of the compound of formula (I)

[0104] Pilot test: About 20 mg of the compound of formula (I) in free form and a certain molar amount of counterion (1.1 eq.) were weighed into a 3.0 mL vial, 1.0 mL of solvent (isopropanol, acetone was selected) was added, and magnetic stirring was carried out at room temperature. After stirring for 2 days, the obtained solid was separated and dried at 50°C under a blast of air, and then XRPD was measured. Two methanesulfonic acid salt forms 1 / 2 were obtained, wherein Form 1 was obtained by using isopropanol as the solvent, and the methanesulfonic acid salt Form 1 had an XRPD pattern substantially as shown in Figure 7 . Form 2 was obtained by using acetone as the solvent Figure 8 . From the XRPD pattern, the crystallinity of the methanesulfonic acid salt Form 1 was slightly better, and isopropanol was selected as the solvent for the later scale-up synthesis of the methanesulfonic acid salt Form 1 of the compound of formula (I).

[0105] Scale-up synthesis: 200.15 mg of the compound of formula (I) in free form (20221231) was weighed into 5 mL of acetone to form a suspension; 5 mL of a solution containing 34.4 μL of methanesulfonic acid (1.1 eq.) in acetone was added under stirring at room temperature; after 4 days of stirring under the cycle of temperature rising and falling, the system was turbid, centrifuged, and dried at 50°C under a blast of air for 2 hours; the solid was crushed, and drying under a blast of air at 50°C was continued overnight. (The cycle of temperature rising and falling was as follows: 40°C (120 min) -> 40°C -> 5°C (0.1°C / min) -> 5°C (120 min) -> 40°C (0.1°C / min) -> 40°C (120 min) -> 5°C (0.1°C / min) -> 5°C (120 min), and after 2 cycles, the temperature was raised to 25°C for constant-temperature stirring).

[0106] The methanesulfonic acid salt Form 1 had an XRPD pattern substantially as shown in Figure 7 . The XRPD pattern of this Form 1 had characteristic diffraction peaks at the following 2θ angles, as shown in Table 3.

[0107] Table 3

[0108]

[0109]

[0110] As shown in Figure 9 , the thermogravimetric analysis (TGA) curve of the methanesulfonic acid salt Form 1 had a weight loss of about 1.34% when the temperature was raised to 120°C, and the sample had thermal stability. The methanesulfonic acid salt Form 1 was an anhydrous substance.

[0111] As shown in Figure 10 , the differential scanning calorimetric analysis (DSC) curve of the methanesulfonic acid salt Form 1 had an endothermic peak at 215.58°C.

[0112] The molar ratio of the compound of formula (I) to hydrochloric acid in methanesulfonic acid Form 2 is about 1:1, which is a salt.

[0113] The methanesulfonic acid Form 2 of 1 The H-NMR spectrum is shown in Figure 11 . From the nuclear magnetic point of view, the compound of this crystal form is structurally indistinguishable from the free base. The salt is about 1:1 of base to acid, and there is no obvious organic solvent residue.

[0114] Example 3: 24-hour equilibrium solubility experiment in water

[0115] The 24-hour equilibrium solubility of the compound of formula (I) in free base form and the methanesulfonic acid salt Form 1 was tested. The specific steps are as follows: 10 mg of sample was weighed into 1 mL of the corresponding solvent to prepare a suspension, and the suspension was stirred magnetically at 37±2℃. After 24 hours, the sample was taken and centrifuged, and the supernatant was filtered and then tested for solubility. The supernatant was filtered through a 0.22 μm PTFE filter membrane and then tested for solubility. The results are shown in Table 4, and the solubility of the compound of formula (I) in free base form in water is about 16.2 μg / mL, and the solubility of the methanesulfonic acid salt Form 1 is 43.07 μg / mL.

[0116] Table 4

[0117] Solid form Equilibrium solubility in water at 37°C (pg / mL) Free base (amorphous) 16.2 Mesylate salt crystalline form 1 43.07

[0118] Example 4: One-week stability experiment

[0119] After the free base and the methanesulfonic acid salt Form 1 and Form 2 samples were placed at 25℃ / 60% RH (long-term) and 40℃ / 75% RH (accelerated) for one week, the HPLC purity and crystal form change Figure 12 ) were tested. The results are shown in Table 5, and the HPLC purity and crystal form of the methanesulfonic acid salt Form 1 did not change significantly after being placed under the two test conditions for one week, indicating that the chemical stability and crystal form stability of the methanesulfonic acid salt Form 1 are both good. The comparison of the spectra is shown in Figure 12 .

[0120] Table 5

[0121]

[0122] Example 5: Hygroscopicity

[0123] The stability risk of the sample with humidity change at 25℃ was evaluated, and the DVS test Figure 13 ) was performed on the methanesulfonic acid salt Form 1 which had better solid state stability, and the solid sample after the test was collected for XRPD test Figure 14 ).

[0124] The results are summarized in Table 6 and Figure 14The methanesulfonate salt Form 1 is slightly hygroscopic. After the test, the crystal form of the sample did not change.

[0125] Table 6 Summary of hygroscopicity evaluation results

[0126]

[0127] *: Hygroscopicity definition in 2020 edition of Chinese Pharmacopoeia: under the condition of 25℃ / 80%RH, the weight gain of the sample is 0.2-2% (less than 2% but not less than 0.2%) for slightly hygroscopic.

Claims

1. A mesylate salt crystalline form of a polycyclic compound, wherein the compound has a structural formula as shown in Formula (I): ###0001### Formula (I) wherein The mesylate salt crystalline form is mesylate salt crystalline form 1, which has an X-ray powder diffraction pattern with characteristic diffraction peaks at 2Θ angles of 4.6±0.2°, 18.3±0.2°, 21.2±0.2°, 25.9±0.2° and 28.0±0.2°.

2. The crystalline form of claim 1, wherein, The X-ray powder diffraction pattern further has one or more characteristic diffraction peaks at 2Θ angles of 4.1±0.2°, 19.8±0.2°, 24.1±0.2° and 26.3±0.2°.

3. The crystalline form of claim 1, wherein, The mesylate salt crystalline form 1 has an XRPD pattern substantially as shown in Figure 5.

4. A pharmaceutical composition comprising the crystalline form of any one of claims 1-3 and a pharmaceutically acceptable carrier.

5. Use of the crystalline form of any one of claims 1-3 or the pharmaceutical composition of claim 4 in the manufacture of a medicament for preventing and / or treating a disease or disorder mediated at least in part by DHX33.

6. Use according to claim 5, wherein, The disease is selected from the group consisting of a cancer, a viral infection and an inflammation mediated by DHX33.

7. A method of preparing the mesylate salt Form 1 of claim 1 comprising: The mesylate salt crystalline form 1 can be obtained by adding isopropanol to the free form of the compound of Formula (I), followed by adding methanesulfonic acid, stirring, collecting the solid and drying. The mesylate salt crystalline form 1 can be obtained by adding isopropanol to the free form of the compound of Formula (I), followed by adding methanesulfonic acid, stirring, collecting the solid and drying.

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