CN116003394A - Hydrochloride crystal form of amide compound, pharmaceutical composition and application - Google Patents

Abstract

The invention belongs to the field of medicinal chemistry and relates to an amide compound, namely 1-(3-cyano-5-methylthiophene-2-yl)-N-(6-methoxyl-1H-benzo[d]imidazole- Different hydrochloride crystal forms of 2-yl)-2,5-dimethyl-1H-pyrrole-3-carboxamide, pharmaceutical compositions and uses comprising the crystal forms. The chemical stability and crystal form stability of the hydrochloride crystal form are better than those of the free alkali crystal form.

Description

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

Technical Field

The present invention belongs to the field of pharmaceutical chemistry and relates to different hydrochloride 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

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 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 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.

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.

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

The object of the present invention is to provide different hydrochloride 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.

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

In a first aspect, the present invention provides a hydrochloride salt form A of a compound of formula (1), whose X-ray powder diffraction (XRPD) pattern has characteristic diffraction peaks at the following 2θ angles: 11.6±0.2°, 22.1±0.2° and 26.0±0.2°.

In some embodiments, the X-ray powder diffraction pattern of the hydrochloride form A of the compound of formula (1) also has characteristic diffraction peaks at one or more of the following 2θ angles: 7.60±0.2°, 8.2±0.2° and 13.0±0.2°.

In some embodiments, the X-ray powder diffraction pattern of the hydrochloride form A of the compound of formula (1) also has characteristic diffraction peaks at one or more of the following 2θ angles: 14.1±0.2°, 16.8±0.2° and 20.7±0.2°.

The present invention also provides a method for preparing the hydrochloride 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 hydrochloric acid, stirring, collecting the solid and then drying to obtain the hydrochloride crystal form A. In a specific embodiment, the organic solvent is isopropanol.

In a second aspect, the present invention provides a hydrochloride 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: 14.65±0.2° and 24.75±0.2°.

In some embodiments, the X-ray powder diffraction pattern of the hydrochloride form B of the compound of formula (1) also has characteristic diffraction peaks at one or more of the following 2θ angles: 11.85±0.2°, 25.18±0.2° and 26.0±0.2°.

In some embodiments, the X-ray powder diffraction pattern of the hydrochloride form B of the compound of formula (1) also has characteristic diffraction peaks at one or more of the following 2θ angles: 19.42±0.2°, 19.80±0.2° and 20.91±0.2°.

The present invention also provides a method for preparing hydrochloride 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 hydrochloric acid, stirring, collecting the solid and then drying to obtain the hydrochloride crystal form B. In a specific embodiment of the present invention, the organic solvent is acetone.

In a third aspect, the present invention provides a hydrochloride salt form C of the compound of formula (1), whose X-ray powder diffraction (XRPD) pattern has characteristic diffraction peaks at the following 2θ angles: 10.95±0.2°, 14.96±0.2° and 19.23±0.2 ^° .

In some embodiments, the X-ray powder diffraction pattern of the hydrochloride crystal form C of the compound of formula (1) also has characteristic diffraction peaks at one or more of the following 2θ angles: 9.04±0.2°, 12.33±0.2° and 26.13±0.2°.

In some embodiments, the X-ray powder diffraction pattern of the hydrochloride crystal form C of the compound of formula (1) also has characteristic diffraction peaks at one or more of the following 2θ angles: 24.86±0.2°, 28.10±0.2° and 31.6±0.2°.

The present invention also provides a method for preparing hydrochloride crystal form C 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 hydrochloric acid, stirring, collecting the solid and then drying to obtain the hydrochloride crystal form B. In a specific embodiment of the present invention, the organic solvent is acetonitrile.

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.

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.

In a fourth 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.

In a fifth 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.

In a sixth 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.

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 only and not limiting of particular embodiments.

Definition of terms

Unless otherwise stated, the technical and scientific terms used herein have the same meanings as those generally understood by 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 lower range limit or preferred values, without considering whether the scope is specifically disclosed. Unless otherwise stated, the numerical range listed herein is intended to include the endpoints of the range and all integers and fractions (decimals) within the range.

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.

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 presence of 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".

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 usually characterized by peak positions (abscissa) and/or peak intensities (ordinate).

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 recording the reflected beam 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.

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.

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.

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.

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.

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

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

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.

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 within the scope of sound medical judgment without excessive toxicity, irritation, allergic reaction or other problems or complications, and are commensurate with a reasonable benefit/risk ratio. "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.

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 inhalants.

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.

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.

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.

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.

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

The dosage frequency of the crystal form 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 form, and then during a period of several days or more days, the patient does not receive a daily dose of the crystal form.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG1 is an X-ray powder diffraction (XRPD) pattern of free crystalline form A of the compound of formula (1).

FIG2 is a thermogravimetric analysis (TGA) diagram of the free crystalline form A of the compound of formula (1).

FIG3 is a differential scanning calorimetry (DSC) diagram of the free crystalline form A of the compound of formula (1).

FIG4 is a ¹ H NMR chart of the free crystalline form A of the compound of formula (1).

FIG5 is an X-ray powder diffraction (XRPD) pattern of the hydrochloride crystal form A of the compound of formula (1).

FIG6 is a thermogravimetric analysis (TGA) diagram of the hydrochloride crystal form A of the compound of formula (1).

FIG. 7 is a differential scanning calorimetry (DSC) diagram of the hydrochloride crystal form A of the compound of formula (1).

FIG8 is a ^{1 H} NMR chart of the hydrochloride crystal form A of the compound of formula (1).

FIG9 is an X-ray powder diffraction (XRPD) pattern of the hydrochloride crystal form B of the compound of formula (1).

FIG10 is a thermogravimetric analysis (TGA) diagram of the hydrochloride crystal form B of the compound of formula (1).

FIG11 is a differential scanning calorimetry (DSC) diagram of the hydrochloride crystal form B of the compound of formula (1).

FIG. 12 is a ^{1 H} NMR chart of Form B of the hydrochloride salt of the compound of formula (1).

FIG. 13 is an X-ray powder diffraction (XRPD) pattern of Form C of the hydrochloride salt of the compound of formula (1).

FIG. 14 is a thermogravimetric analysis (TGA) diagram of the hydrochloride crystal form C of the compound of formula (1).

FIG15 is a differential scanning calorimetry (DSC) diagram of the hydrochloride crystal form C of the compound of formula (1).

FIG. 16 is a ^{1 H} NMR chart of Form C of the hydrochloride salt of the compound of formula (1).

DETAILED DESCRIPTION

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.

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.

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

X-ray powder diffraction (XRPD)

XRPD Testing Parameters

Thermogravimetric analysis (TGA) and differential scanning calorimetry (DSC)

TGA and DSC test parameters

Nuclear magnetic spectrum ( ^1H NMR)

¹ H NMR test parameters

Ion Chromatography (IC)

IC Test Parameters

High Performance Liquid Chromatography (HPLC)

HPLC test parameters

Dynamic Water Sorption Analysis (DVS)

DVS test parameters

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

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

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.

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

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.

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

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 chromatography (Prep-HPLC) (acetonitrile/water (containing 0.1% formic acid)) to obtain free crystalline form A of the compound of formula (I) (5 mg, yield: 4%). MS (ESI) m/z: 406 [M+H ⁺ ]. ¹ H NMR (400 MHz, DMSO-d ₆ ): δ 12.04 (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=7.2 Hz, 1H), 3.73 (s, 3H), 2.47 (s, 3H), 2.38 (s, 3H), 2.04 (s, 3H).

The free form Form A has an XRPD pattern substantially as shown in FIG1 .

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

Table 1

As shown in FIG. 2 , 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.

As shown in FIG3 , the differential scanning calorimetry (DSC) curve of the free crystalline form A has an endothermic peak at about 163° C.

The ¹ H NMR spectrum of the free crystalline form A of the compound of formula (1) is shown in FIG4 .

Example 2: Hydrochloride salt of the compound of formula (I) and its preparation method

Preparation method of hydrochloride crystal form A of compound of formula (I)

Weigh about 200 mg of the free state sample (2734020-1) into a 20 mL glass vial; add 10 mL of isopropanol to dissolve the sample (vortex and ultrasound promote sample dissolution); add 60.06 mg of hydrochloric acid (hydrochloric acid mass content: 36% to 38%), add a stirrer, and stir at room temperature on a magnetic stirrer; after stirring at room temperature for about 2 days, filter to obtain a solid, and place it in a vacuum drying oven at 60°C for 12 hours. The hydrochloride crystal form A has an XRPD pattern substantially as shown in Figure 5.

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

Table 2-1

As shown in Figure 6, the thermogravimetric analysis (TGA) curve of hydrochloride crystal form A shows a weight loss of about 0.3390% when the temperature is raised to about 150°C. Hydrochloride crystal form A is an anhydrous substance.

As shown in FIG. 7 , the differential scanning calorimetry (DSC) curve of hydrochloride salt form A has an endothermic peak at about 233.42° C.

The ¹ H-NMR spectrum of hydrochloride salt form A is shown in FIG8 .

Preparation method of hydrochloride crystal form B of compound of formula (I)

Weigh about 200 mg of the free sample (2734020-1) into a 20 mL glass vial; add 5 mL of acetone to dissolve the sample (vortex and ultrasound promote sample dissolution); add 60.06 mg of hydrochloric acid (hydrochloric acid mass content: 36% to 38%), add a stirrer, and suspend and stir at room temperature on a magnetic stirrer; after stirring at room temperature for about 2 days, filter out the solid, and place it in a vacuum drying oven at 60°C to dry for 12 hours.

The hydrochloride salt Form B has an XRPD pattern substantially as shown in FIG. 9 .

The XRPD pattern of hydrochloride crystal form B has characteristic diffraction peaks at the following 2θ angles, as shown in Table 2-2:

Table 2-2

As shown in Figure 10, the thermogravimetric analysis (TGA) curve of hydrochloride crystal form B shows a weight loss of about 4.7791% when the temperature is raised to about 140°C. Hydrochloride crystal form B is a hydrate or anhydrous form but has water adsorbed on the surface.

As shown in FIG. 11 , the differential scanning calorimetry (DSC) curve of hydrochloride salt form B has multiple endothermic peaks at about 31-114° C. and melts at 205.98° C.

In the hydrochloride salt form B, the molar ratio of the compound of formula (I) to hydrochloric acid is about 1:1.

The ¹ H-NMR spectrum of hydrochloride salt form B is shown in FIG12 .

Preparation method of hydrochloride crystal form C of compound of formula (I)

Weigh about 200 mg of the free sample (2734020-1) into a 20 mL glass vial; add 5 mL of acetonitrile to dissolve the sample (vortex and ultrasound promote sample dissolution); add 60.06 mg of hydrochloric acid (hydrochloric acid mass content: 36% to 38%), add a stirrer, and suspend and stir at room temperature on a magnetic stirrer; after stirring at room temperature for about 2 days, filter out the solid, and place it in a vacuum drying oven at 60°C to dry for 12 hours.

Hydrochloride salt Form C has an XRPD pattern substantially as shown in Figure 13.

The XRPD pattern of hydrochloride crystal form C has characteristic diffraction peaks at the following 2θ angles, as shown in Table 2-3:

Table 2-3

As shown in Figure 14, the thermogravimetric analysis (TGA) curve of hydrochloride crystal form C shows a weight loss of about 4.7791% when the temperature is raised to about 140°C. Hydrochloride crystal form C is a hydrate or anhydrous form but has water adsorbed on the surface.

As shown in FIG. 15 , the differential scanning calorimetry (DSC) curve of hydrochloride salt form C has multiple endothermic peaks at about 31-114° C. and melts at 205.98° C.

In the hydrochloride salt form C, the molar ratio of the compound of formula (I) to hydrochloric acid is about 1:1.

The ¹ H-NMR spectrum of hydrochloride salt form C is shown in FIG16 .

Example 3: 24-hour equilibrium solubility experiment in water

The 24-hour equilibrium solubility of free base crystal form A and hydrochloride crystal form A was tested. The specific steps are: weigh 10 mg of free base crystal form A and hydrochloride 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 hydrochloride crystal form A is 1.0 μg/mL.

Table 3

Solid form Equilibrium solubility in water at 37℃ (μg/mL) Free base form A 1.92 Hydrochloride Form A 1.0

Example 4: One-week stability experiment

Free base crystal form A and hydrochloride 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 hydrochloride crystal form A was placed under the two test conditions for one week, HPLC purity and crystal form did not change significantly, and the chemical stability and crystal form stability were both good.

Table 4

Example 5: Hygroscopicity test

To evaluate the stability risk of the sample with humidity changes at 25°C, a DVS test was performed on the hydrochloride form C. The results are summarized in Table 5. The sample is slightly hygroscopic.

After the DVS test, the crystal form of the hydrochloride form C samples did not undergo significant changes.

Table 5

Claims (9)

1. A hydrochloride crystalline form of an amide compound, wherein the amide compound has a structural formula shown in formula (1):

2. the crystalline form of claim 1, which is hydrochloride form a, having an X-ray powder diffraction pattern with characteristic diffraction peaks at the following 2Θ angles: 11.6.+ -. 0.2 ^° 22.1±0.2° and 26.0±0.2°;

preferably, its X-ray powder diffraction pattern also has characteristic diffraction peaks at one or more of the following 2θ angles: 7.60±0.2°, 8.2±0.2° and 13.0±0.2°;

preferably, its X-ray powder diffraction pattern also has characteristic diffraction peaks at one or more of the following 2θ angles: 14.1±0.2°, 16.8±0.2° and 20.7±0.2°;

preferably, the hydrochloride salt form a has an XRPD pattern substantially as shown in figure 5.

3. The crystalline form of claim 1, which is hydrochloride form B, having an X-ray powder diffraction pattern with characteristic diffraction peaks at the following 2Θ angles: 14.65±0.2° and 24.75±0.2°;

preferably, its X-ray powder diffraction pattern also has characteristic diffraction peaks at one or more of the following 2θ angles: 11.85±0.2°, 25.18±0.2° and 26.0±0.2°;

preferably, its X-ray powder diffraction pattern also has characteristic diffraction peaks at one or more of the following 2θ angles: 19.42±0.2°, 19.80±0.2° and 20.91±0.2°;

preferably, the hydrochloride salt form B has an XRPD pattern substantially as shown in figure 9.

4. The crystalline form of claim 1, which is hydrochloride form C, having an X-ray powder diffraction pattern with characteristic diffraction peaks at the following 2Θ angles: 10.95±0.2°, 14.96±0.2° and 19.23±0.2°;

preferably, its X-ray powder diffraction pattern also has characteristic diffraction peaks at one or more of the following 2θ angles: 9.04±0.2°, 12.33±0.2° and 26.13±0.2°;

preferably, its X-ray powder diffraction pattern also has characteristic diffraction peaks at one or more of the following 2θ angles: 24.86±0.2°, 28.10±0.2° and 31.6±0.2°;

preferably, the hydrochloride form C has an XRPD pattern substantially as shown in figure 13.

5. A pharmaceutical composition comprising a crystalline form according to any one of claims 1-4 and a pharmaceutically acceptable carrier for use in the prevention and/or treatment of a disease or disorder mediated at least in part by DHX33, preferably the disease is selected from the group consisting of DHX33 mediated cancer, viral infection and inflammation.

6. Use of a crystalline form according to any one of claims 1-4 or a pharmaceutical composition according to claim 5 in the manufacture of a medicament for the prevention and/or treatment of a disease or disorder mediated at least in part by DHX33, preferably selected from the group consisting of DHX33 mediated cancer, viral infection and inflammation.

7. A process for the preparation of form a of the hydrochloride salt of claim 2 comprising: adding an organic solvent into the free crystal form A of the compound of the formula (1), then adding hydrochloric acid, stirring, collecting solid and then drying to obtain the hydrochloride crystal form A, wherein the organic solvent is preferably isopropanol.

8. A process for the preparation of form B of the hydrochloride salt of claim 3 comprising: adding an organic solvent into the free crystal form A of the compound of the formula (1), then adding hydrochloric acid, stirring, collecting the solid, and then drying to obtain the hydrochloride crystal form B, wherein the organic solvent is preferably acetone.

9. A process for the preparation of form C of the hydrochloride salt of claim 4 comprising: adding an organic solvent into the free crystal form A of the compound of the formula (1), then adding hydrochloric acid, stirring, collecting the solid, and then drying to obtain the hydrochloride crystal form C, wherein the organic solvent is acetonitrile.

Images