CN111499632A

CN111499632A - Crystal form I containing alkynyl compound, preparation method and application thereof

Info

Publication number: CN111499632A
Application number: CN202010623632.9A
Authority: CN
Inventors: 温剑锋; 林艳琼; 冯建鹏; 吴天助; 邵振中; 李卫东
Original assignee: Guangzhou Shunjian Biomedicine Technology Co ltd; Suzhou Yasheng Pharmaceutical Co ltd; Yasheng Pharmaceutical Group Hong Kong Co ltd; SHANGHAI YASHENG MEDICAL TECHNOLOGY CO LTD
Current assignee: Guangzhou Shunjian Biomedicine Technology Co ltd; Suzhou Yasheng Pharmaceutical Co ltd; Yasheng Pharmaceutical Group Hong Kong Co ltd; SHANGHAI YASHENG MEDICAL TECHNOLOGY CO LTD; Shanghai Ascentage Pharmaceutical Co Ltd
Priority date: 2020-07-02
Filing date: 2020-07-02
Publication date: 2020-08-07
Anticipated expiration: 2040-07-02
Also published as: CN111499632B

Abstract

The invention discloses a crystal form I of an alkynyl-containing compound, and a preparation method and application thereof. The invention specifically discloses a crystal form I of 3- ((1H-pyrazolo [3,4-b ] pyridine-5-substituted) ethynyl) -4-methyl-N- (4- ((4-methylpiperazine-1-substituted) methyl) -3- (trifluoromethyl) phenyl) benzamide. The crystal form of the invention has good stability and has important value for the optimization and development of medicaments.

Description

Crystal form I containing alkynyl compound, preparation method and application thereof

Technical Field

The invention belongs to the field of chemical medicine, and particularly relates to a crystal form I of an alkynyl-containing compound, and a preparation method and application thereof.

Background

The application relates to an alkynyl-containing compound with a chemical name of 3- ((1H-pyrazolo [3,4-b ] pyridine-5-substituted) ethynyl) -4-methyl-N- (4- ((4-methylpiperazine-1-substituted) methyl) -3- (trifluoromethyl) phenyl) benzamide, and the patent has limited reports on the compound and does not relate to the crystal form of the compound.

Given the common polymorphism of compounds, a general drug may have two or more different crystal states. The existence form and the quantity of the polymorphic compound are unpredictable, and different crystal forms of the same medicament have obvious difference in the aspects of solubility, melting point, density, stability and the like, so that the temperature form, the uniformity, the bioavailability, the curative effect and the safety of the medicament are influenced to different degrees. Therefore, comprehensive polymorphic form screening of the compound is required in the process of developing a new drug, and the selection of a crystal form suitable for developing a pharmaceutical preparation has important clinical significance.

Disclosure of Invention

The invention provides a crystal form I of an alkynyl-containing compound, and a preparation method and application thereof. The crystal form of the invention has good stability and has important value for the optimization and development of medicaments.

The invention provides a crystal form I of 3- ((1H-pyrazolo [3,4-b ] pyridine-5-substituted) ethynyl) -4-methyl-N- (4- ((4-methylpiperazine-1-substituted) methyl) -3- (trifluoromethyl) phenyl) benzamide, which has characteristic peaks at 9.498 +/-0.2 degrees, 12.293 +/-0.2 degrees, 13.045 +/-0.2 degrees, 15.899 +/-0.2 degrees, 16.199 +/-0.2 degrees, 18.183 +/-0.2 degrees, 18.327 +/-0.2 degrees, 21.755 +/-0.2 degrees, 22.362 +/-0.2 degrees and 25.69 +/-0.2 degrees in an X-ray powder diffraction pattern represented by 2 theta angles.

In certain preferred embodiments of the present invention, the crystalline form I, which has an X-ray powder diffraction pattern expressed in terms of 2 Θ angle, has characteristic peaks at 8.968 ± 0.2 °, 9.498 ± 0.2 °, 12.293 ± 0.2 °, 13.045 ± 0.2 °, 15.899 ± 0.2 °, 16.199 ± 0.2 °, 16.533 ± 0.2 °, 16.908 ± 0.2 °, 18.183 ± 0.2 °, 18.327 ± 0.2 °, 20.042 ± 0.2 °, 20.271 ± 0.2 °, 21.755 ± 0.2 °, 22.362 ± 0.2 °, 25.69 ± 0.2 °.

In certain preferred embodiments of the present invention, the crystalline form I, having an X-ray powder diffraction pattern expressed in terms of 2 Θ angles, has 2 Θ values as shown in table 1;

TABLE 1

。

In certain preferred embodiments of the present invention, the form I has an X-ray powder diffraction pattern expressed in terms of 2 Θ angles substantially as shown in figure 1. The thermogravimetric analysis (TGA) of form I has a weight loss gradient of 0.15% at 200 ℃, the "%" is a weight percentage, and the TGA profile is preferably substantially as shown in figure 2. In the differential scanning thermal spectrum (DSC) of the crystal form I, a thermal absorption peak exists at 235 ℃, and the DSC spectrum is preferably basically as shown in figure 3. The dynamic moisture sorption pattern (DVS) of form I is substantially as shown in figure 4. In the polarization microscope picture of the crystal form I, the crystal form is in a flaky crystal, and the polarization microscope picture is preferably basically as shown in FIG. 5.

In the present invention, the X-ray powder diffraction is performed by using K α ray.

In the present invention, the target type used in the X-ray powder diffraction is a Cu target.

The invention also provides 3- ((1H-pyrazole [3, 4-b)]A process for preparing crystalline form I of pyridine-5-substituted) ethynyl) -4-methyl-N- (4- ((4-methylpiperazin-1-substituted) methyl) -3- (trifluoromethyl) phenyl) benzamide comprising the steps of: in an organic solvent, adding 3- ((1H-pyrazolo [3, 4-b)]Pyridine-5-substituted) ethynyl) -4-methyl-N- (4- ((4-methylpiperazine-1-substituted) methyl) -3- (trifluoromethyl) phenyl) benzamide is crystallized to obtain 3- ((1H-pyrazolo [3, 4-b)]The crystal form I of pyridine-5-substituted) ethynyl) -4-methyl-N- (4- ((4-methylpiperazine-1-substituted) methyl) -3- (trifluoromethyl) phenyl) benzamide is shown, and the organic solvent is C₁-C₁₀An alkane solvent of (C)₁-C₄One or more of alcohol solvents, ether solvents, nitrile solvents, ketone solvents, ester solvents and DMSO.

In the preparation method of the crystal form I, the crystallization method can be a method well known in the art, such as suspension stirring, stirring at normal temperature, crystallization by heating and cooling, solvent volatilization method or anti-solvent addition method.

In the preparation method of the crystal form I, the 3- ((1H-pyrazolo [3,4-b ] pyridine-5-substituted) ethynyl) -4-methyl-N- (4- ((4-methylpiperazine-1-substituted) methyl) -3- (trifluoromethyl) phenyl) benzamide is prepared by a patent reference method.

In the preparation method of the crystal form I, the organic solvent is preferably one or more of heptane, methanol, ethanol, isopropanol, methyl tert-butyl ether, acetonitrile, acetone, 2-butanone, ethyl acetate, isopropyl acetate and DMSO.

In the preparation method of the crystalline form I, the mass-to-volume ratio of the 3- ((1H-pyrazolo [3,4-b ] pyridine-5-substituted) ethynyl) -4-methyl-N- (4- ((4-methylpiperazin-1-substituted) methyl) -3- (trifluoromethyl) phenyl) benzamide to the organic solvent may be a mass-to-volume ratio conventional in the art, preferably 1:1-1:5g/m L, more preferably 1:1-1:3 g/m L, such as 1:2.5 g/m L.

In the preparation method of the crystal form I, the temperature of crystallization can be the conventional temperature in the field, such as 20-50 ℃.

In the preparation method of the crystal form I, the crystallization time is not particularly limited, and the crystal I can be precipitated, for example, for 1 to 36 hours, for example, for 1 to 5 hours, and for example, for 1 to 3 hours.

In the preparation method of the crystal form I, when the crystal adopts a normal-temperature stirring method, the organic solvent is preferably C₁-C₁₀An alkane solvent of (C)₁-C₄And one or more of an alcohol solvent, a nitrile solvent, a ketone solvent, an ether solvent and an ester solvent, more preferably one or more of heptane, methanol, ethanol, isopropanol, acetonitrile, acetone, 2-butanone, methyl tert-butyl ether, isopropyl acetate and ethyl acetate.

In the preparation method of the crystal form I, when the crystal adopts an anti-solvent addition method, the organic solvent is preferably DMSO.

In the preparation method of the crystal form I, when the crystallization adopts an anti-solvent addition method, the anti-solvent is preferably one or more of water, an alcohol solvent and a nitrile solvent, the water can be one or more of distilled water, deionized water, purified water, tap water and mineral water, the alcohol solvent is preferably isopropanol, the nitrile solvent is preferably acetonitrile, the mass volume ratio of the 3- ((1H-pyrazolo [3,4-b ] pyridine-5-substituted) ethynyl) -4-methyl-N- (4- ((4-methylpiperazine-1-substituted) methyl) -3- (trifluoromethyl) phenyl) benzamide to the anti-solvent can be the conventional mass volume ratio in the field, preferably 1:2-1:25g/m L, more preferably 1:2-1:20g/m L, such as 1:2.5 g/m L, 1:7.5 g/m L or 1:20g/m L.

The preparation method of the crystal form I preferably comprises the following steps: 3- ((1H-pyrazole [3, 4-b)]Mixing pyridine-5-substituted) ethynyl) -4-methyl-N- (4- ((4-methylpiperazine-1-substituted) methyl) -3- (trifluoromethyl) phenyl) benzamide with an organic solvent, stirring, and filtering to obtain a target crystal form; the organic solvent is C₁-C₁₀An alkane solvent of (C)₁-C₄Preferably one or more of alcohols, nitriles, ketones, ethers and esters, preferably heptane, methanol, ethanol, isopropyl alcoholOne or more of alcohol, acetonitrile, acetone, 2-butanone, methyl tert-butyl ether, isopropyl acetate and ethyl acetate. The stirring is preferably carried out at 20 to 50 ℃. The stirring is preferably carried out for 1 to 5 hours. After the filtration is finished, drying is preferably included. The drying is preferably vacuum drying. The 3- ((1H-pyrazole [3, 4-b)]The mass-to-volume ratio of pyridine-5-substituted) ethynyl) -4-methyl-N- (4- ((4-methylpiperazine-1-substituted) methyl) -3- (trifluoromethyl) phenyl) benzamide to the organic solvent is preferably 1:1 to 1:5g/m L.

The preparation method of the crystal form I preferably comprises the following steps of mixing 3- ((1H-pyrazolo [3,4-b ] pyridine-5-substituted) ethynyl) -4-methyl-N- (4- ((4-methylpiperazine-1-substituted) methyl) -3- (trifluoromethyl) phenyl) benzamide with DMSO, adding an antisolvent, and filtering to obtain the target crystal form, wherein the antisolvent is one or more of water, an alcohol solvent and a nitrile solvent, preferably one or more of water, isopropanol and acetonitrile, the addition amount of the antisolvent is preferably determined by precipitation of a large amount of solid, the addition manner of the antisolvent is preferably slow, the mass-volume ratio of the 3- ((1H-pyrazolo [3,4-b ] pyridine-5-substituted) ethynyl) -4-methyl-N- (4- ((4-methylpiperazine-1-substituted) methyl) -3- (trifluoromethyl) phenyl) benzamide to the DMSO is preferably 1:1-1:5g/m L, and the mass-volume ratio of the 3- ((1H-pyrazolo [3, 4-substituted) methyl) -3- (trifluoromethyl) phenyl) benzamide to the DMSO is preferably 1: 5-substituted methyl-ethynyl-3- (1: 25: 84-trifluoromethyl) benzamide.

The invention also provides a pharmaceutical composition, which comprises the crystal form I and pharmaceutically acceptable auxiliary materials. The crystalline form I can be in a therapeutically effective amount. The pharmaceutically acceptable excipients may be those well known in the art, and in the case of solid formulations, include, but are not limited to: diluents, binders, disintegrants, lubricants, glidants, release rate controlling agents, plasticizers, preservatives, antioxidants and the like.

The pharmaceutical composition can be selected from dosage forms suitable for human administration, such as: tablet, capsule, granule, powder, pill, etc., preferably tablet, capsule, granule, disintegrating tablet, sustained release tablet or controlled release tablet.

The pharmaceutical composition of the present invention can be prepared by methods well known in the art, and can be prepared into dosage forms suitable for human administration by mixing a therapeutically effective amount of the crystalline form I with various pharmaceutical excipients, such as: the tablet, capsule and granule can be prepared by mixing, granulating, tabletting or encapsulating.

The invention also provides an application of the crystal form I or the pharmaceutical composition in preparing medicines. The medicament is preferably a medicament for preventing and/or treating cancer. The cancer includes but is not limited to one or more of gastrointestinal stromal tumor, histiocytic lymphoma, non-small cell lung cancer, pancreatic cancer, breast cancer, prostate cancer, liver cancer, skin cancer, epithelial cell cancer, nasopharyngeal carcinoma and leukemia. The medicament preferably comprises a therapeutically effective amount of the above-mentioned crystalline form I, or the above-mentioned pharmaceutical composition.

The above preferred conditions can be arbitrarily combined to obtain preferred embodiments of the present invention without departing from the common general knowledge in the art.

The reagents and starting materials used in the present invention are commercially available.

The positive progress effects of the invention are as follows:

1. the crystal form I of 3- ((1H-pyrazolo [3,4-b ] pyridine-5-substituted) ethynyl) -4-methyl-N- (4- ((4-methylpiperazine-1-substituted) methyl) -3- (trifluoromethyl) phenyl) benzamide is not reported in the prior art, and the application discovers multiple novel crystal forms of the compound for the first time. Through a large number of experiments and screening, the crystal form I is prepared for the first time and is used as a candidate object.

2. The crystal form I prepared by the invention has good stability, is convenient to store, can avoid the risk of crystal transformation in the process of drug development or production, avoids the change of bioavailability and drug effect, can be developed into a dosage form suitable for clinical use, and has very high economic value.

3. The invention also provides a preparation method of the crystal form I, which is simple and convenient to operate, high in reproducibility, not easy to leave solvent, environment-friendly and suitable for different large-scale production.

Drawings

Figure 1 is an XPRD pattern of crystalline form I of 3- ((1H-pyrazolo [3,4-b ] pyridine-5-substituted) ethynyl) -4-methyl-N- (4- ((4-methylpiperazin-1-substituted) methyl) -3- (trifluoromethyl) phenyl) benzamide.

Figure 2 is a TGA diagram of crystalline form I of 3- ((1H-pyrazolo [3,4-b ] pyridine-5-substituted) ethynyl) -4-methyl-N- (4- ((4-methylpiperazin-1-substituted) methyl) -3- (trifluoromethyl) phenyl) benzamide.

Figure 3 is a DSC diagram of crystalline form I of 3- ((1H-pyrazolo [3,4-b ] pyridine-5-substituted) ethynyl) -4-methyl-N- (4- ((4-methylpiperazin-1-substituted) methyl) -3- (trifluoromethyl) phenyl) benzamide.

Figure 4 is a DVS diagram of crystalline form I of 3- ((1H-pyrazolo [3,4-b ] pyridine-5-substituted) ethynyl) -4-methyl-N- (4- ((4-methylpiperazin-1-substituted) methyl) -3- (trifluoromethyl) phenyl) benzamide.

Figure 5 is a micrograph of crystalline form I of 3- ((1H-pyrazolo [3,4-b ] pyridine-5-substituted) ethynyl) -4-methyl-N- (4- ((4-methylpiperazin-1-substituted) methyl) -3- (trifluoromethyl) phenyl) benzamide.

Figure 6 is an XPRD pattern of 3- ((1H-pyrazolo [3,4-b ] pyridine-5-substituted) ethynyl) -4-methyl-N- (4- ((4-methylpiperazin-1-substituted) methyl) -3- (trifluoromethyl) phenyl) benzamide monohydrochloride form III.

Figure 7 is an XPRD pattern of crystalline form IV of 3- ((1H-pyrazolo [3,4-b ] pyridine-5-substituted) ethynyl) -4-methyl-N- (4- ((4-methylpiperazin-1-substituted) methyl) -3- (trifluoromethyl) phenyl) benzamide dihydrochloride.

Figure 8 is an XPRD diagram of 3- ((1H-pyrazolo [3,4-b ] pyridine-5-substituted) ethynyl) -4-methyl-N- (4- ((4-methylpiperazin-1-substituted) methyl) -3- (trifluoromethyl) phenyl) benzamide sulfate crystalline form VI.

Figure 9 is an XPRD diagram of 3- ((1H-pyrazolo [3,4-b ] pyridine-5-substituted) ethynyl) -4-methyl-N- (4- ((4-methylpiperazin-1-substituted) methyl) -3- (trifluoromethyl) phenyl) benzamide phosphate crystalline form VII.

Figure 10 is an XPRD pattern of 3- ((1H-pyrazolo [3,4-b ] pyridine-5-substituted) ethynyl) -4-methyl-N- (4- ((4-methylpiperazin-1-substituted) methyl) -3- (trifluoromethyl) phenyl) benzamide citrate crystalline form VIII.

Figure 11 is an XPRD pattern of 3- ((1H-pyrazolo [3,4-b ] pyridine-5-substituted) ethynyl) -4-methyl-N- (4- ((4-methylpiperazin-1-substituted) methyl) -3- (trifluoromethyl) phenyl) benzamide mesylate salt form X.

Figure 12 is an XPRD pattern of crystalline form a of 3- ((1H-pyrazolo [3,4-b ] pyridine-5-substituted) ethynyl) -4-methyl-N- (4- ((4-methylpiperazin-1-substituted) methyl) -3- (trifluoromethyl) phenyl) benzamide.

Detailed Description

The invention is further illustrated by the following examples, which are not intended to limit the scope of the invention. The experimental methods without specifying specific conditions in the following examples were selected according to the conventional methods and conditions, or according to the commercial instructions.

The present invention will be described in further detail with reference to specific examples. It is to be understood that these embodiments are provided to illustrate the basic principles, essential features and advantages of the present invention, and the present invention is not limited by the following embodiments. The implementation conditions used in the examples can be further adjusted according to specific requirements, and the implementation conditions not indicated are generally the conditions in routine experiments.

In the following examples, the experimental procedures are generally carried out according to conventional conditions or according to conventional test conditions, and the compounds may be obtained by organic synthesis or by commercially available methods. The compounds used in the following examples were obtained by a commercially available method with a purity of 99%.

The abbreviations used in the present invention are explained as follows:

XPRD-X ray powder diffraction

TGA-thermogravimetric analysis

DSC-differential scanning calorimetry

DVS-dynamic moisture desorption analysis

P L M-polarization microscope analysis

The test conditions were as follows:

XRPD

the solids were characterized using an X-ray powder diffractometer (bruker D8 advance or D2 Phase).

Scanning angle: 3 ° (2 θ) -40 ° (2 θ).

Step length: 0.02 ° (2 θ).

Scanning speed: 0.3 sec/step (D8), 0.2 sec/step (D2).

Voltage of light pipe: 40KV (D8), 30 KV (D2).

Light pipe current: 40 mA (D8) and 10 mA (D2).

Rotating: and opening.

Sample pan: zero background sample pan.

Thermogravimetric analysis Q500 or Discovery TGA 55 was used for the thermogravimetric analysis of solid samples using TA Instrument. After equilibrating the sample trays, the samples were hung on a wire and lifted into the oven. After stabilization, the samples were heated at a rate of 10 ℃/min to different end point temperatures.

DSC analysis of solid samples was performed using a TA Instrument differential scanning calorimeter Q200 and Discovery DSC 250. The samples were weighed and the values recorded, and then placed in the sample chamber. The samples were heated from 25 ℃ to different end temperatures at a rate of 10 ℃/min.

DVS analysis of the solids was performed using an IGAsorp dynamic water sorption instrument.

Temperature: at 25 ℃.

Airflow: 250 m L/min.

And (3) scanning circulation: 2.

the shortest test time: and (3) 30 min.

The longest test time: and 2 h.

Waiting for balance: 98 percent.

The samples were observed using a Nikon Eclipse L V100N PO L type polarizing microscope.

Example 1: preparation of form I

40mg of 3- ((1H-pyrazolo [3,4-b ] pyridine-5-substituted) ethynyl) -4-methyl-N- (4- ((4-methylpiperazin-1-substituted) methyl) -3- (trifluoromethyl) phenyl) benzamide was added to 0.1m L methanol, stirred at room temperature for 1-3H, filtered and the solid collected to give crystalline form I.

The XPRD pattern of form I is shown in figure 1; in the X-ray powder diffraction pattern expressed by the angle 2 theta, the value 2 theta is shown in Table 1;

TABLE 1

。

The TGA profile of form I is shown in figure 2 with a weight loss gradient of 0.15% at 200 ℃, where "%" is weight percent. The DSC chart of the crystal form I is shown in figure 3, and the crystal form I has a heat absorption peak at 235 ℃; the DVS profile of form I is shown in figure 4; the microscopic image of the crystal form I is shown in FIG. 5, and the crystal form is crystallized in a sheet structure.

Example 2: preparation of form I

40mg of 3- ((1H-pyrazolo [3,4-b ] pyridine-5-substituted) ethynyl) -4-methyl-N- (4- ((4-methylpiperazin-1-substituted) methyl) -3- (trifluoromethyl) phenyl) benzamide was added to 0.1m L ethanol, stirred at room temperature for 1-3H, filtered and the solid collected to give crystalline form I.

Example 3: preparation of form I

40mg of 3- ((1H-pyrazolo [3,4-b ] pyridine-5-substituted) ethynyl) -4-methyl-N- (4- ((4-methylpiperazin-1-substituted) methyl) -3- (trifluoromethyl) phenyl) benzamide was added to 0.1m L isopropanol, stirred at room temperature for 1-3H, filtered and the solid collected to give crystalline form I.

Example 4: preparation of form I

40mg of 3- ((1H-pyrazolo [3,4-b ] pyridine-5-substituted) ethynyl) -4-methyl-N- (4- ((4-methylpiperazin-1-substituted) methyl) -3- (trifluoromethyl) phenyl) benzamide was added to 0.1m L methyl tert-butyl ether, stirred at room temperature for 1-3H, filtered and the solid collected to give crystalline form I.

Example 5: preparation of form I

40mg of 3- ((1H-pyrazolo [3,4-b ] pyridine-5-substituted) ethynyl) -4-methyl-N- (4- ((4-methylpiperazin-1-substituted) methyl) -3- (trifluoromethyl) phenyl) benzamide was added to 0.1m L2-butanone, stirred at room temperature for 1-3H, filtered and the solid collected to give crystalline form I.

Example 6: preparation of form I

40mg of 3- ((1H-pyrazolo [3,4-b ] pyridine-5-substituted) ethynyl) -4-methyl-N- (4- ((4-methylpiperazin-1-substituted) methyl) -3- (trifluoromethyl) phenyl) benzamide was added to 0.1m L acetonitrile, stirred at room temperature for 1-3H, filtered and the solid collected to give form I.

Example 7: preparation of form I

40mg of 3- ((1H-pyrazolo [3,4-b ] pyridine-5-substituted) ethynyl) -4-methyl-N- (4- ((4-methylpiperazin-1-substituted) methyl) -3- (trifluoromethyl) phenyl) benzamide was added to 0.1m L acetone, stirred at room temperature for 1-3H, filtered and the solid collected to give crystalline form I.

Example 8: preparation of form I

40mg of 3- ((1H-pyrazolo [3,4-b ] pyridine-5-substituted) ethynyl) -4-methyl-N- (4- ((4-methylpiperazin-1-substituted) methyl) -3- (trifluoromethyl) phenyl) benzamide was added to 0.1m L isopropyl acetate, stirred at room temperature for 1-3H, filtered and the solid collected to give crystalline form I.

Example 9: preparation of form I

40mg of 3- ((1H-pyrazolo [3,4-b ] pyridine-5-substituted) ethynyl) -4-methyl-N- (4- ((4-methylpiperazin-1-substituted) methyl) -3- (trifluoromethyl) phenyl) benzamide was added to 0.1m L methanol, heated to 50 ℃ and stirred for 1-5H, then cooled to room temperature, filtered and the solid was collected to give form I.

Example 10: preparation of form I

The same reaction conditions as those in example 1 are adopted, the solvent is replaced by heptane or ethyl acetate, and after solid is precipitated, the mixture is stirred for 1 to 36 hours at the temperature of between 20 and 50 ℃ to obtain the crystal form I.

Example 11: preparation of form I

40mg of 3- ((1H-pyrazolo [3,4-b ] pyridine-5-substituted) ethynyl) -4-methyl-N- (4- ((4-methylpiperazin-1-substituted) methyl) -3- (trifluoromethyl) phenyl) benzamide was added to 0.1m L DMSO, stirred at room temperature for 1-3H, slowly added with 0.1m L water, filtered and the solid collected to give form I.

Example 12: preparation of form I

40mg of 3- ((1H-pyrazolo [3,4-b ] pyridine-5-substituted) ethynyl) -4-methyl-N- (4- ((4-methylpiperazine-1-substituted) methyl) -3- (trifluoromethyl) phenyl) benzamide was added to 0.1m L DMSO, stirred at room temperature for 1-3H, slowly added to 0.3m L acetonitrile, filtered and the solid collected to give form I.

Example 13: preparation of form I

40mg of 3- ((1H-pyrazolo [3,4-b ] pyridine-5-substituted) ethynyl) -4-methyl-N- (4- ((4-methylpiperazin-1-substituted) methyl) -3- (trifluoromethyl) phenyl) benzamide was added to 0.1m L DMSO, stirred at room temperature for 1-3H, slowly added 0.8m L isopropanol, filtered and the solid collected to give form I.

The XPRD pattern, TGA pattern, DSC pattern, DVS pattern and microscopic pattern of form I prepared in examples 2-13 above are substantially the same as form I prepared in example 1.

Example 14: stability testing of form I (different temperature, humidity and light)

Stability studies were performed on form I prepared in examples 1-13, and prior to testing, form I had less than 0.05% single impurities and less than 0.05% total impurities.

Placing the crystal form I at 60 ℃, high humidity and illumination conditions, sampling at 0 day/5 day/10 days, and inspecting the content and related substances of the crystal form I, wherein the illumination conditions are that the total illumination is more than or equal to 1.2 × 10⁶L ux hr, near ultraviolet energy not less than 200w hr/m². The results are shown in Table 2.

TABLE 2

The result shows that the content and the purity of the crystal form I are almost unchanged, the content is close to 100%, the single impurity content is less than 0.05%, the total impurity content is less than 0.05%, and the crystal form I shows better stability when the crystal form I is sampled and measured for 5 days and 10 days under the conditions of 60 ℃, high humidity and illumination.

Example 15: stability tests (different solvents) were carried out on the crystalline modification I prepared in examples 1 to 13

Weighing a sample crystal form I, placing the sample crystal form I in a sample bottle, adding a solvent to prepare a suspension, stirring the obtained suspension at room temperature and 50 ℃ for 3 days, filtering, collecting a solid, and characterizing the solid. The results are shown in table 3 below.

Table 3 suspension stirring test of form I

As can be seen from the above table, form I has good stability in a variety of solvents.

Example 16: hygroscopicity test of form I

The hygroscopicity studies were carried out on the crystalline modification I prepared in examples 1 to 13, and 10mg of the crystalline modification I were subjected to a dynamic moisture sorption (DVS) test. The conclusion is as described in table 4 below:

TABLE 4

The above shows that the crystal form I is not easy to absorb moisture in the storage process, is easy to store and can have a longer shelf life.

Example 17: preparation of form A

Preparation of form a: dissolving 3- ((1H-pyrazolo [3,4-b ] pyridine-5-substituted) ethynyl) -4-methyl-N- (4- ((4-methylpiperazine-1-substituted) methyl) -3- (trifluoromethyl) phenyl) benzamide in ethyl acetate, heating to 50 ℃ to enable the mixture to reach a saturated state, filtering, stirring at a low speed at room temperature, separating out a solid in the ethyl acetate, namely filtering and collecting the solid to obtain the crystal form A. In an X-ray powder diffraction pattern of the crystal form A expressed by 2 theta angles, the 2 theta values are shown in a table 5;

TABLE 5 2 theta values for form A

。

The X-ray powder diffraction pattern of the crystal form A expressed by the angle of 2 theta is shown in figure 12.

Example 18: competitive studies were carried out on forms I and A prepared in examples 1-13

1.2mg of the crystal form I and 1.2mg of the crystal form A are mixed in equal amount, then 0.2m L ethanol-water (volume ratio is 1:1) mixed solvent is added, the obtained suspension is stirred for 5 days at room temperature, and XRPD result shows that the crystal form I is stable crystal form.

Example 19: crystalline form III of 3- ((1H-pyrazolo [3,4-b ] pyridine-5-substituted) ethynyl) -4-methyl-N- (4- ((4-methylpiperazine-1-substituted) methyl) -3- (trifluoromethyl) phenyl) benzamide monohydrochloride

Adding 40mg of 3- ((1H-pyrazolo [3,4-b ] pyridine-5-substituted) ethynyl) -4-methyl-N- (4- ((4-methylpiperazine-1-substituted) methyl) -3- (trifluoromethyl) phenyl) benzamide into a system with 0.2m of L methanol and dichloromethane (the volume ratio of the methanol to the dichloromethane is 1:1), stirring at room temperature, adding 1.05 equivalents of hydrochloric acid to completely dissolve the solid, adding ethyl acetate, continuing to stir for 0.5H, separating out the solid, continuing to stir for 4H, filtering, and vacuum-drying at 50 ℃ overnight to obtain the crystal form III.

In an X-ray powder diffraction pattern of the crystal form III expressed by 2 theta angles, the 2 theta value is shown in a table 6;

TABLE 62 theta values for form III

The XPRD pattern of form III is shown in figure 6.

Example 20: preparation of form IV

Adding 40mg of 3- ((1H-pyrazolo [3,4-b ] pyridine-5-substituted) ethynyl) -4-methyl-N- (4- ((4-methylpiperazine-1-substituted) methyl) -3- (trifluoromethyl) phenyl) benzamide into a system containing 0.4m of L methanol and dichloromethane (the volume ratio of the methanol to the dichloromethane is 1:1), stirring at room temperature, adding 2.1 equivalents of hydrochloric acid to precipitate a solid, continuously stirring for 1-2H, filtering, and vacuum-drying at 50 ℃ overnight to obtain the crystal form IV.

In an X-ray powder diffraction pattern of the crystal form IV expressed by 2 theta angles, the 2 theta values are shown in a table 7;

TABLE 7 2 theta values of form IV

The XPRD pattern of form IV is shown in figure 7.

Example 21: preparation of form VI

40mg of 3- ((1H-pyrazolo [3,4-b ] pyridine-5-substituted) ethynyl) -4-methyl-N- (4- ((4-methylpiperazin-1-substituted) methyl) -3- (trifluoromethyl) phenyl) benzamide was added to 0.4m L of methanol and dichloromethane (1: 1), stirred at room temperature, 1 equivalent of sulfuric acid was added, the solid was dissolved, stirring was continued, ethyl acetate (2 m L) was added to produce a solid precipitate, which was slowly cooled to room temperature, stirred overnight, filtered, and vacuum dried overnight at 50 ℃ to obtain crystalline form VI.

In an X-ray powder diffraction pattern of the crystal form VI expressed by 2 theta angles, the 2 theta values are shown in a table 8;

table 8 2 theta values for form VI

The XPRD pattern of form VI is shown in figure 8.

Example 22: preparation of form VII

Adding 40mg of 3- ((1H-pyrazolo [3,4-b ] pyridine-5-substituted) ethynyl) -4-methyl-N- (4- ((4-methylpiperazine-1-substituted) methyl) -3- (trifluoromethyl) phenyl) benzamide into 1m L ethanol, heating to 50 ℃, stirring, adding 1 equivalent of phosphoric acid, dissolving the solid, continuously stirring to generate a solid precipitate, continuously stirring at 50 ℃ for 0.5H, then reducing to room temperature, stirring for 2H, filtering, and vacuum-drying at 50 ℃ overnight to obtain the crystal form VII.

In an X-ray powder diffraction pattern of the crystal form VII expressed by 2 theta angles, the 2 theta values are shown in a table 9;

TABLE 9 2 θ values for form VII

The XPRD pattern of form VII is shown in figure 9.

Example 23: preparation of form VIII

40mg of 3- ((1H-pyrazolo [3,4-b ] pyridine-5-substituted) ethynyl) -4-methyl-N- (4- ((4-methylpiperazine-1-substituted) methyl) -3- (trifluoromethyl) phenyl) benzamide was added to 0.5m L of THF, heated to 60 ℃ and stirred, 1 equivalent of citric acid was added, stirring was continued, a solid immediately appeared, filtered, and vacuum dried overnight at 50 ℃ to give form VIII.

In an X-ray powder diffraction pattern of the crystal form VIII expressed by 2 theta angles, the 2 theta values are shown in a table 10;

TABLE 10 2 θ values for form VIII

The XPRD pattern of form VIII is shown in figure 10.

Example 24: preparation of form X

Adding 40mg of 3- ((1H-pyrazolo [3,4-b ] pyridine-5-substituted) ethynyl) -4-methyl-N- (4- ((4-methylpiperazine-1-substituted) methyl) -3- (trifluoromethyl) phenyl) benzamide into 1m L ethanol, stirring at 50 ℃, adding 1.05 equivalents of methanesulfonic acid, dissolving the solid, slowly cooling to room temperature, stirring for 2H, filtering, and vacuum-drying at 50 ℃ overnight to obtain the crystal form X.

In an X-ray powder diffraction pattern of the crystal form X expressed by a 2 theta angle, 2 theta values are shown in Table 11;

TABLE 11 2 θ values for form X

The XPRD pattern of form X is shown in figure 11.

Example 25: comparative study of other crystalline forms

The obtained spectrum of the crystal form was analyzed by comparison, and the analysis results are shown in table 12 below.

TABLE 12

As can be seen from the above table, form I has good properties in terms of thermal stability, hygroscopicity and crystallinity. Form I has better thermal stability relative to form III and form VI; the crystal form I has more excellent performance in hygroscopicity compared with the crystal form VII and the crystal form X; the crystal form I has no obvious polycrystal and is more excellent compared with the crystal forms III, IV, VII and VIII; form I has better crystallinity relative to form IV and form VIII.

Claims

1. A crystal form I of 3- ((1H-pyrazolo [3,4-b ] pyridine-5-substituted) ethynyl) -4-methyl-N- (4- ((4-methylpiperazine-1-substituted) methyl) -3- (trifluoromethyl) phenyl) benzamide has characteristic peaks at 9.498 +/-0.2 °, 12.293 +/-0.2 °, 13.045 +/-0.2 °, 15.899 +/-0.2 °, 16.199 +/-0.2 °, 18.183 +/-0.2 °, 18.327 +/-0.2 °, 21.755 +/-0.2 °, 22.362 +/-0.2 ° and 25.69 +/-0.2 ° in an X-ray powder diffraction pattern represented by 2 theta angles.

2. A crystalline form I of 3- ((1H-pyrazolo [3,4-b ] pyridine-5-substituted) ethynyl) -4-methyl-N- (4- ((4-methylpiperazin-1-substituted) methyl) -3- (trifluoromethyl) phenyl) benzamide according to claim 1 characterized in that it has a characteristic peak at 8.968 ± 0.2 °, 9.498 ± 0.2 °, 12.293 ± 0.2 °, 13.045 ± 0.2 °, 15.899 ± 0.2 °, 16.199 ± 0.2 °, 16.533 ± 0.2 °, 16.908 ± 0.2 °, 18.183 ± 0.2 °, 18.327 ± 0.2 °, 20.042 ± 0.2 °, 20.271 ± 0.2 °, 21.755 ± 0.2 °, 22.362 ± 0.2 °, 25.69 ± 0.2 ° in its X-ray powder diffraction pattern expressed in terms of 2 Θ angles.

3. Crystalline form I of 3- ((1H-pyrazolo [3,4-b ] pyridine-5-substituted) ethynyl) -4-methyl-N- (4- ((4-methylpiperazin-1-substituted) methyl) -3- (trifluoromethyl) phenyl) benzamide according to claim 2 characterized in that it has the following 2 Θ values in an X-ray powder diffractogram expressed in terms of 2 Θ angles:

；

and/or, in the thermogravimetric analysis pattern of the crystal form I, the weight loss gradient at 200 ℃ is 0.15%;

and/or a heat absorption peak is existed at 235 ℃ in the differential scanning heat spectrum of the crystal form I.

4. The crystalline form I of 3- ((1H-pyrazolo [3,4-b ] pyridine-5-substituted) ethynyl) -4-methyl-N- (4- ((4-methylpiperazin-1-substituted) methyl) -3- (trifluoromethyl) phenyl) benzamide of claim 3 wherein the X-ray powder diffraction pattern of form I expressed in terms of 2 Θ angles is as shown in figure 1;

and/or the thermogravimetric analysis pattern of the crystal form I is shown in figure 2;

and/or, the differential scanning thermal spectrum of form I is shown in figure 3;

and/or the dynamic moisture adsorption profile of form I is shown in figure 4.

5. A preparation method of the crystal form I of 3- ((1H-pyrazolo [3,4-b ] pyridine-5-substituted) ethynyl) -4-methyl-N- (4- ((4-methylpiperazine-1-substituted) methyl) -3- (trifluoromethyl) phenyl) benzamide as claimed in any one of claims 1 to 4, comprising the steps of crystallizing 3- ((1H-pyrazolo [3,4-b ] pyridine-5-substituted) ethynyl) -4-methyl-N- (4- ((4-methylpiperazine-1-substituted) methyl) -3- (trifluoromethyl) phenyl) benzamide in an organic solvent to obtain the crystal form I of 3- ((1H-pyrazolo [3,4-b ] pyridine-5-substituted) ethynyl) -4-methyl-N- (4- ((4-methylpiperazine-1-substituted) methyl) -3- (trifluoromethyl) phenyl) benzamide, wherein the organic solvent is heptane, methanol, ethanol, isopropanol, methyl butanone, acetonitrile, acetone, 2-isopropyl acetate, ethyl acetate, dimethylsulfoxide, dimethyl sulfoxide, or dimethyl sulfoxide, stirring at normal temperature, stirring at which the temperature is lower than the temperature, stirring at which one or more than the volume of the organic solvent is equal to 20H, and adding water to the crystal form I of 3- ((1-methyl-pyrazole [ 3- (4-methyl) benzamide, wherein the crystal form I is obtained by stirring at normal temperature, and the temperature is equal to 25H.

6. The method according to claim 5, wherein the mass-to-volume ratio of the 3- ((1H-pyrazolo [3,4-b ] pyridine-5-substituted) ethynyl) -4-methyl-N- (4- ((4-methylpiperazin-1-substituted) methyl) -3- (trifluoromethyl) phenyl) benzamide to the organic solvent is from 1:1 to 1:3 g/m L;

and/or the crystallization time is 1-5 h;

and/or, when the crystallization adopts an anti-solvent addition method, the anti-solvent is one or more of water, an alcohol solvent and a nitrile solvent, and the water is one or more selected from distilled water, deionized water, purified water, tap water and mineral water;

and/or, when the crystallization adopts an anti-solvent addition method, and the anti-solvent is one or more of water, an alcohol solvent and a nitrile solvent, the alcohol solvent is isopropanol;

and/or, when the crystallization adopts an anti-solvent addition method, and the anti-solvent is one or more of water, an alcohol solvent and a nitrile solvent, the nitrile solvent is acetonitrile;

and/or, when the crystallization adopts an anti-solvent addition method, and the anti-solvent is one or more of water, an alcohol solvent and a nitrile solvent, the mass-to-volume ratio of the 3- ((1H-pyrazolo [3,4-b ] pyridine-5-substituted) ethynyl) -4-methyl-N- (4- ((4-methylpiperazine-1-substituted) methyl) -3- (trifluoromethyl) phenyl) benzamide to the anti-solvent is 1:2-1:25g/m L.

7. The process according to claim 6, wherein the crystallization time is 1 to 3 hours;

and/or, when the crystallization adopts an anti-solvent addition method, and the anti-solvent is one or more of water, an alcohol solvent and a nitrile solvent, the mass-to-volume ratio of the 3- ((1H-pyrazolo [3,4-b ] pyridine-5-substituted) ethynyl) -4-methyl-N- (4- ((4-methylpiperazine-1-substituted) methyl) -3- (trifluoromethyl) phenyl) benzamide to the anti-solvent is 1:2-1:20g/m L.

8. A pharmaceutical composition comprising the crystalline form I according to any one of claims 1 to 4, and a pharmaceutically acceptable excipient.

9. Use of the crystalline form I according to any one of claims 1 to 4, or the pharmaceutical composition according to claim 8, for the preparation of a medicament for the prevention and/or treatment of cancer.

10. The use of claim 9, wherein the cancer is selected from one or more of gastrointestinal stromal tumor, histiocytic lymphoma, non-small cell lung cancer, pancreatic cancer, breast cancer, prostate cancer, liver cancer, skin cancer, epithelial cell cancer, nasopharyngeal cancer and leukemia.