CN111732586A

CN111732586A - Crystal form of alkynyl-containing compound salt, preparation method and application

Info

Publication number: CN111732586A
Application number: CN202010634264.8A
Authority: CN
Inventors: 温剑锋; 林艳琼; 冯建鹏; 吴天助; 邵振中; 李卫东
Original assignee: Suzhou Yasheng Pharmaceutical Co ltd; Yasheng Pharmaceutical Group Hong Kong Co ltd; Guangzhou Shunjian Biomedicine Technology Co ltd
Current assignee: Healthquest Pharma Inc; Suzhou Yasheng Pharmaceutical Co ltd; Yasheng Pharmaceutical Group Hong Kong Co ltd
Priority date: 2020-07-02
Filing date: 2020-07-02
Publication date: 2020-10-02
Anticipated expiration: 2040-07-02
Also published as: CN111732586B

Abstract

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

Description

Crystal form of alkynyl-containing compound salt, preparation method and application

Technical Field

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

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 of alkynyl-containing compound salt, a preparation method and application. 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 II of 3- ((1H-pyrazolo [3,4-b ] pyridine-5-substituted) ethynyl) -4-methyl-N- (4- ((4-methylpiperazine-1-substituted) methyl) -3- (trifluoromethyl) phenyl) benzamide fumarate, which has characteristic peaks at 11.817 +/-0.2 degrees, 13.984 +/-0.2 degrees, 15.434 +/-0.2 degrees, 17.853 +/-0.2 degrees, 18.89 +/-0.2 degrees, 19.825 +/-0.2 degrees and 21.718 +/-0.2 degrees in an X-ray powder diffraction pattern represented by a 2 theta angle;

or characteristic peaks are found at 4.461 + -0.2 °, 11.817 + -0.2 °, 13.251 + -0.2 °, 13.984 + -0.2 °, 15.434 + -0.2 °, 16.78 + -0.2 °, 17.853 + -0.2 °, 18.89 + -0.2 °, 19.825 + -0.2 °, 21.718 + -0.2 °, 22.056 + -0.2 °, 24.652 + -0.2 °, 25.198 + -0.2 ° and 26.762 + -0.2 °;

or characteristic peaks are found at 4.461 + -0.2 °, 8.904 + -0.2 °, 11.817 + -0.2 °, 12.244 + -0.2 °, 13.251 + -0.2 °, 13.6 + -0.2 °, 13.984 + -0.2 °, 15.434 + -0.2 °, 15.9 + -0.2 °, 16.78 + -0.2 °, 17.154 + -0.2 °, 17.853 + -0.2 °, 18.89 + -0.2 °, 19.825 + -0.2 °, 20.926 + -0.2 °, 21.718 + -0.2 °, 22.056 + -0.2 °, 22.656 + -0.2 °, 24.35 + -0.2 °, 24.652 + -0.2 °, 25.198 + -0.2 °, 25.88 + -0.2 °, 26.301 + -0.2 °, 26.762 + -0.2 °, 27.836 + -0.2 ° and 28.179 + -0.2 °.

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

TABLE 1

Angle of diffraction	Relative strength	d value	Angle of diffraction	Relative strength	d value
						(2θ°)	(％)	(Angstrom)	(2θ°)	(％)	(Angstrom)
4.461	41.4	19.79333	24.35	27.1	3.6525
						8.904	20	9.924	24.652	48.1	3.60846
11.817	80.6	7.48317	25.198	48.2	3.5315
						12.244	24.3	7.22324	25.88	25.7	3.43995
13.251	45.8	6.67645	26.301	26.8	3.38583
						13.6	34.1	6.50553	26.762	45.4	3.32852
13.984	50.6	6.32792	27.836	23	3.20251
						15.434	71.2	5.73649	28.179	22.6	3.16424
15.9	19.9	5.56949	28.65	11.2	3.11328
						16.78	44.1	5.27934	29.855	12.1	2.99033
17.154	36.3	5.16509	30.494	14.1	2.92915
						17.853	100	4.96424	30.607	15.3	2.91859
18.89	50.3	4.69408	31.677	13.5	2.82236
						19.825	50.5	4.47468	32.656	11.7	2.73999
20.926	28.2	4.24173	36.767	10.8	2.44247
						21.718	54.2	4.08883	37.328	10.1	2.40703
22.056	42.5	4.02688	37.984	8.8	2.367
						22.656	23.5	3.92152	38.408	10.1	2.34181
23.282	14.7	3.81762	39.149	10.1	2.29918
						23.732	17.5	3.74614

。

In certain preferred embodiments of the present invention, the crystalline form II has an X-ray powder diffraction pattern expressed in terms of 2-theta angles substantially as shown in figure 1. The TGA profile of form II has a weight loss gradient of 0.31% at 200 ℃ and the "%" is weight percent, and the TGA profile is substantially as shown in figure 2. The DSC pattern of the crystal form II has a heat absorption peak at 251 ℃, and the DSC pattern is preferably basically as shown in figure 3. This peak should be the melting peak of the sample and decomposition immediately follows melting. The DVS pattern of form II, which is substantially as shown in figure 4, preferably has a moisture uptake of 1.66% at 90% Relative Humidity (RH) and no change in the XRPD pattern of the sample. It is known that fumarate has better solid form and properties. In the polarization microscope picture of the crystal form II, the crystal form is granular crystals, and the polarization microscope picture is preferably basically as shown in FIG. 5.

In the present invention, the ray used for the X-ray powder diffraction is a Ka ray.

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

The invention also provides a method for preparing the crystalline form II of 3- ((1H-pyrazolo [3,4-b ] pyridine-5-substituted) ethynyl) -4-methyl-N- (4- ((4-methylpiperazine-1-substituted) methyl) -3- (trifluoromethyl) phenyl) benzamide fumarate, comprising the steps of: in an organic solvent, 3- ((1H-pyrazolo [3,4-b ] pyridine-5-substituted) ethynyl) -4-methyl-N- (4- ((4-methylpiperazine-1-substituted) methyl) -3- (trifluoromethyl) phenyl) benzamide reacts with fumaric acid, and crystallization is performed to obtain the crystal form II 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 fumarate.

In the preparation method of the crystal form II, the crystallization method may 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 II, 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 II, the organic solvent can be a conventional organic solvent in the field, and can also be one or more (such as C) of alkane solvents, alcohol solvents, ketone solvents, ester solvents, aromatic hydrocarbon solvents, halogenated hydrocarbon solvents, nitrile solvents, ether solvents, aliphatic hydrocarbon solvents, DMF (dimethyl formamide) and DMSO (dimethyl sulfoxide)₁-C₁₀An alkane solvent of (C)₁-C₄One or more of acetone, 2-butanone, ethyl acetate, isopropyl acetate, toluene, dichloromethane, dichloroethane, chloroform, acetonitrile, diethyl ether, methyl tert-butyl ether, tetrahydrofuran, cyclohexane, DMF, and DMSO), preferably one or more of an alcoholic solvent, a halogenated hydrocarbon solvent, and an ether solvent, more preferably one or more of methanol, ethanol, isopropanol, dichloromethane, and tetrahydrofuran.

In the preparation method of the crystalline form II, 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 to 1:25g/mL, more preferably 1:5 to 1:25g/mL, such as 1:5g/mL, 1:10g/mL, 1:12.5g/mL or 1:25 g/mL.

In the preparation method of the crystalline form II, the molar ratio of the fumaric acid to the 3- ((1H-pyrazolo [3,4-b ] pyridine-5-substituted) ethynyl) -4-methyl-N- (4- ((4-methylpiperazin-1-substituted) methyl) -3- (trifluoromethyl) phenyl) benzamide may be a molar ratio conventional in the art, preferably 1:0.9 to 1:3, such as 1:0.9, 1:1, 1:1.05 or 1: 2.1.

In the preparation method of the crystal form II, the temperature of crystallization can be the conventional temperature in the field, such as 20-60 ℃.

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

In the preparation method of the crystal form II, when the crystallization adopts an anti-solvent addition method, the anti-solvent is preferably an ester solvent, and more preferably ethyl acetate. The mass-to-volume ratio of the compound to the antisolvent is not particularly limited, and crystal precipitation is not affected.

The preparation method of the crystal form II preferably comprises the following steps: 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 an organic solvent, adding fumaric acid, and filtering to obtain the target crystal form. The organic solvent is preferably one or more of an alcohol solvent, a halogenated hydrocarbon solvent and an ether solvent, and more preferably one or more of methanol, ethanol, isopropanol, dichloromethane and tetrahydrofuran. The mixing and adding of the fumaric acid are preferably carried out under stirring. After the filtration is finished, drying is preferably included, the drying is preferably vacuum drying, and the drying temperature is preferably 40-60 ℃, such as 50 ℃.

The invention also provides a pharmaceutical composition, which comprises the crystal form II and pharmaceutically acceptable auxiliary materials. The crystalline form II 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 II 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 II 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 II, 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 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 II is prepared for the first time and is used as a candidate object.

2. The crystal form II prepared by the invention has good stability and convenient storage, 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 strong economic value.

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

Drawings

Figure 1 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 fumarate crystalline form II.

Figure 2 is a TGA 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 fumarate crystalline form II.

Figure 3 is a DSC 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 fumarate crystalline form II.

Figure 4 is a DVS 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 fumarate crystalline form II.

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

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.

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

PLM-polarizing 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: 0.3sec/step (D8), 0.2sec/step (D2).

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

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

Rotating: and opening.

Sample pan: zero background sample pan.

TGA

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

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

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

Temperature: at 25 ℃.

Airflow: 250 mL/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.

PLM

The samples were observed using a Nikon Eclipse LV100N POL type polarization microscope.

Example 1: preparation of form II

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 0.5mL of methanol and dichloromethane (the volume ratio of methanol to dichloromethane was 1:1), stirred at room temperature, 1 equivalent of fumaric acid was added, solid immediately appeared, stirring was continued overnight, filtered, and dried under vacuum at 50 ℃ overnight to give crystalline form II. The XPRD pattern of form II 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 II, wherein the weight loss gradient at 200 ℃ is 0.31%, and the "%" is weight percent, is shown in figure 2. The DSC pattern of the crystal form II has a heat absorption peak at 251 ℃, and the DSC pattern is shown in figure 3. This peak should be the melting peak of the sample and decomposition immediately follows melting. The DVS pattern for form II, which had a moisture uptake of 1.66% at 90% Relative Humidity (RH), was unchanged by the XRPD pattern of the sample, and is shown in figure 4. It is known that fumarate has better solid form and properties. In the polarization microscope picture of the crystal form II, the crystal form is granular crystal, and the polarization microscope picture is shown in figure 5.

Example 2: 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 0.2mL of methanol and dichloromethane (the volume ratio of the methanol to the dichloromethane is 1:1), mixing the mixture into a system, stirring the mixture 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 the mixture, and performing vacuum drying at 50 ℃ for 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 2;

TABLE 2 values of 2 theta for form III

Angle of diffraction	Relative strength	d value	Angle of diffraction	Relative strength	d value
						(2θ°)	(％)	(Angstrom)	(2θ°)	(％)	(Angstrom)
5.397	100	16.36285	23.615	41.7	3.76449
						8.27	20.6	10.68294	23.828	40.3	3.73127
10.703	29.8	8.25938	24.861	12.5	3.57858
						11.213	10.5	7.88469	25.31	17.8	3.51603
13.561	20.1	6.5241	26.44	72.6	3.36833
						13.861	11.4	6.38369	26.871	16.8	3.31527
15.27	10.1	5.79766	27.924	17.6	3.19254
						15.635	19.6	5.66331	28.767	12	3.10089
16.097	22.9	5.50165	29.837	6.7	2.99214
						16.374	21.4	5.40916	30.56	10	2.92293
16.938	17.3	5.23026	31.016	12.7	2.88104
						18.411	12.5	4.81501	32.066	6.9	2.78902
18.86	11.8	4.70146	33.556	12.6	2.66846
						19.46	20.7	4.55792	34.28	7	2.61376
20.501	21.7	4.32873	36.069	5.1	2.48816
						21.041	61.3	4.21873	37.291	6.9	2.40937
21.514	26.8	4.12717	38.063	6.8	2.36224
						22.298	28.4	3.98365	38.726	6	2.32331
22.601	21.6	3.93096

。

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

Example 3: 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 0.4mL of 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 a 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 3;

TABLE 3 2 theta values of form IV

Angle of diffraction	Relative strength	d value	Angle of diffraction	Relative strength	d value
						(2θ°)	(％)	(Angstrom)	(2θ°)	(％)	(Angstrom)
4.293	100	20.56553	16.629	56.3	5.3268
						5.147	79.2	17.15621	16.797	58.5	5.27394
6.242	87.4	14.14906	17.224	79.7	5.14416
						9.021	41.5	9.79522	18.501	48.8	4.79187
10.709	47.3	8.25469	19.024	49.3	4.66139
						11.898	62.1	7.43193	21.692	61	4.09373
12.896	83.3	6.85926	25.508	79.4	3.48917
						14.333	65.5	6.17468	28.342	45.4	3.14644
14.65	61.5	6.04155	29.629	36.6	3.01267
						15.648	47.5	5.65838

。

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

Example 4: preparation of form VI

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 to 0.4mL of methanol and dichloromethane (1: 1), stirring at room temperature, adding 1 equivalent of sulfuric acid, dissolving the solid, continuing to stir, adding ethyl acetate (2mL) to generate a solid precipitate, slowly cooling to room temperature, stirring overnight, filtering, and vacuum-drying at 50 ℃ overnight to obtain the crystal form VI.

In an X-ray powder diffraction pattern of the crystal form VI expressed by 2 theta angles, the 2 theta value is shown in a table 4;

TABLE 4 2 theta values for form VI

Angle of diffraction	Relative strength	d value	Angle of diffraction	Relative strength	d value
						(2θ°)	(％)	(Angstrom)	(2θ°)	(％)	(Angstrom)
4.365	91.9	20.22792	18.872	25.8	4.69845
						8.031	25.8	10.99964	19.114	30.7	4.6396
8.309	38.8	10.63289	19.621	26.3	4.52083
						10.357	100	8.53397	19.97	33.9	4.44257
12.253	18.7	7.21787	20.762	25.9	4.27493
						13.036	39.2	6.78585	21.253	47.7	4.17725
13.548	28.2	6.53064	22.358	29.4	3.97315
						13.899	17.5	6.36631	23.251	27.9	3.82251
14.566	40.3	6.07637	24.272	32.1	3.66405
						15.323	22.8	5.77783	24.646	37.3	3.60933
15.518	27.2	5.70574	25.05	29.7	3.55196
						16.055	29.7	5.51602	25.474	26.8	3.49378
16.7	29.4	5.30436	26.292	24.9	3.3869
						17.463	49.6	5.07421	26.727	24.2	3.33284
18.292	36.8	4.84621	29.136	14.8	3.06248

。

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

Example 5: 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 1mL of 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 5;

TABLE 52 theta values of form VII

Angle of diffraction	Relative strength	d value	Angle of diffraction	Relative strength	d value
						(2θ°)	(％)	(Angstrom)	(2θ°)	(％)	(Angstrom)
3.178	39.6	27.78238	20.982	11	4.23055
						7.206	18.6	12.25751	21.307	25.1	4.16667
8.51	32.1	10.38204	21.984	22.8	4.03998
						9.56	9.1	9.24387	22.657	12.1	3.92143
10.637	49.8	8.31024	23.148	12	3.83928
						12.741	40.3	6.94248	23.974	94.4	3.70886
13.197	12.5	6.70324	24.848	14	3.58037
						13.606	26.8	6.50306	25.784	21.5	3.45248
14.097	29.9	6.27738	26.785	43	3.32569
						14.471	43.6	6.11586	27.756	13.8	3.21154
15.144	55.3	5.84562	28.211	10.3	3.16072
						15.939	7.9	5.55582	28.593	9.8	3.11936
16.52	19.7	5.36179	29.726	11.6	3.00301
						17.056	46.5	5.19455	30.498	10.7	2.92878
17.543	17.2	5.05133	32.317	7.1	2.76793
						17.904	10.7	4.95039	33.603	6.8	2.66486
19.043	100	4.65668	36.459	5.4	2.46243
						20.245	14.9	4.38286	37.825	5.8	2.37656

。

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

Example 6: preparation of form VIII

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 to 0.5mL of THF, heating to 60 ℃ and stirring, adding 1 equivalent of citric acid, continuing stirring, immediately appearing a solid, filtering, and vacuum drying at 50 ℃ overnight to obtain the crystalline form VIII.

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

TABLE 62 theta values for form VIII

Angle of diffraction	Relative strength	d value	Angle of diffraction	Relative strength	d value
						(2θ°)	(％)	(Angstrom)	(2θ°)	(％)	(Angstrom)
4.455	100	19.82056	15.481	10.2	5.71922
						6.694	8.9	13.19358	16.296	11.8	5.43481
8.297	27.1	10.64757	16.588	11.5	5.3398
						8.6	16.7	10.27356	17.172	18.7	5.15969
8.883	15.5	9.94707	17.892	17.4	4.95363
						10.036	8.6	8.80676	19.163	13	4.6279
10.886	13.7	8.12114	20.008	16.6	4.43414
						11.743	13.5	7.52999	21.651	13.5	4.10127
12.138	16	7.28565	22.517	9.4	3.9455
						12.856	33.8	6.88041	25.9	8.3	3.43736
13.307	54	6.64819	26.436	10.5	3.36882
						14.614	18.7	6.05634	27.76	7.8	3.21102
15.126	12.3	5.85277	29.377	5	3.0379

。

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

Example 7: 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 1mL of 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, the 2 theta value is shown in a table 7;

TABLE 7 2 theta values for form X

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

Effect example 1: stability testing of fumarate salt form II (different temperature, humidity)

Stability studies were performed on form II prepared in example 1, with less than 0.05% single impurities and less than 0.05% total impurities, before testing.

Placing the crystal form II at 60 ℃ under high humidity condition, sampling at 0 day/5 day/10 days, and inspecting the content and related substances thereof under the illumination condition that the total illumination is more than or equal to 1.2 × 10⁶Lux hr, near ultraviolet energy not less than 200w hr/m². The results are shown in Table 8.

TABLE 8

Crystal form II	Content (wt.)	Simple impurity	Total miscellaneous	XPRD
					Day
0	98.70％	＜0.05％	＜0.05％	Crystal form II
					5 days-high humidity	99.40％	＜0.05％	＜0.05％	Crystal form II
5 days-60 deg.C	99.50％	＜0.05％	＜0.05％	Crystal form II
					10 days-high humidity	99.00％	＜0.05％	＜0.05％	Crystal form II
10 days to 60 DEG C	99.20％	＜0.05％	＜0.05％	Crystal form II

。

The result shows that the content and the purity of the crystal form II are almost unchanged, the content can reach 98.5 percent or even more than 90 percent, the single impurity content is less than 0.05 percent, the total impurity content is less than 0.05 percent, and the crystal form II shows better stability when being sampled and measured for 5 days and 10 days under the high-humidity condition at 60 ℃.

Effect example 2: stability study of Crystal form II (different solvents)

Weighing a sample crystal form II in a sample bottle, adding a solvent to prepare a suspension, stirring the obtained suspension for 3-4 days at room temperature and 50 ℃, filtering and collecting a solid, and performing vacuum drying at room temperature to characterize the solid. The results are shown in Table 9.

Table 9 crystal form II suspension stirring test

Serial number	Solvent(s)	Suspension at room temperature	Suspension at 50 deg.C
				1	Methanol	Crystal form II	Crystal form II
2	Ethanol	Crystal form II	Crystal form II
				3	Isopropanol (I-propanol)	Crystal form II	Crystal form II
4	Acetonitrile	Crystal form II	Crystal form II
				5	Ethyl acetate	Crystal form II	Crystal form II
6	Acetic acid isopropyl ester	Crystal form II	N/A
				7	Tetrahydrofuran (THF)	Crystal form II	Crystal form II
8	N-heptane	Crystal form II	N/A
				9	1, 4-dioxane	Crystal form II	N/A
10	Methanol to water 3: 1 (v: v)	Crystal form II	N/A

。

Note: the above N/A indicates no measurement.

As can be seen from the above table, form II has good stability at both room temperature and 50 ℃.

Effect example 3: study on hygroscopicity of Crystal form II

The hygroscopicity study was performed using crystalline form II prepared in example 1, and about 10mg of crystalline form II was subjected to a dynamic moisture sorption (DVS) test. The conclusion are set forth in table 10 below:

watch 10

Free base/salt	DVS(90％RH)	XRPD before and after DVS
			Crystal form II	1.66％	Is not changed

。

The results show that the crystal form II is not easy to absorb moisture in the storage process, is easy to store and can prolong the shelf life.

Effect example 4: comparative study of form II with other forms

The obtained spectrum of the crystal form was analyzed by comparison, and the analysis results are shown in table 11 below.

TABLE 11

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

Claims

1. Crystalline form II of 3- ((1H-pyrazolo [3,4-b ] pyridine-5-substituted) ethynyl) -4-methyl-N- (4- ((4-methylpiperazin-1-substituted) methyl) -3- (trifluoromethyl) phenyl) benzamide fumarate having characteristic peaks at 11.817 ± 0.2 °, 13.984 ± 0.2 °, 15.434 ± 0.2 °, 17.853 ± 0.2 °, 18.89 ± 0.2 °, 19.825 ± 0.2 ° and 21.718 ± 0.2 ° in its X-ray powder diffraction pattern, expressed in terms of 2 Θ angles;

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

angle of diffraction Relative strength Angle of diffraction Relative strength Angle of diffraction Relative strength (2θ°) (％) (2θ°) (％) (2θ°) (％) 4.461 41.4 19.825 50.5 27.836 23 8.904 20 20.926 28.2 28.179 22.6 11.817 80.6 21.718 54.2 28.65 11.2 12.244 24.3 22.056 42.5 29.855 12.1 13.251 45.8 22.656 23.5 30.494 14.1 13.6 34.1 23.282 14.7 30.607 15.3 13.984 50.6 23.732 17.5 31.677 13.5 15.434 71.2 24.35 27.1 32.656 11.7 15.9 19.9 24.652 48.1 36.767 10.8 16.78 44.1 25.198 48.2 37.328 10.1 17.154 36.3 25.88 25.7 37.984 8.8 17.853 100 26.301 26.8 38.408 10.1 18.89 50.3 26.762 45.4 39.149 10.1

；

And/or in the thermogravimetric analysis pattern of the crystal form II, the weight loss gradient at 200 ℃ is 0.33 percent, and the percent is weight percent;

and/or a heat absorption peak is existed at 246.6 ℃ in the differential scanning heat spectrum of the crystal form II.

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

and/or the thermogravimetric analysis pattern of the crystalline form II is substantially as shown in figure 2;

and/or, the differential scanning calorimetry spectrum of form II is substantially as shown in FIG. 3;

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

4. A process for the preparation of crystalline form II of 3- ((1H-pyrazolo [3,4-b ] pyridine-5-substituted) ethynyl) -4-methyl-N- (4- ((4-methylpiperazin-1-substituted) methyl) -3- (trifluoromethyl) phenyl) benzamide fumarate as claimed in any one of claims 1 to 3 comprising the steps of: in an organic solvent, 3- ((1H-pyrazolo [3,4-b ] pyridine-5-substituted) ethynyl) -4-methyl-N- (4- ((4-methylpiperazine-1-substituted) methyl) -3- (trifluoromethyl) phenyl) benzamide reacts with fumaric acid, and crystallization is performed to obtain the crystal form II 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 fumarate.

5. The preparation method according to claim 4, wherein the crystallization method is suspension stirring, stirring at normal temperature, crystallization by heating and cooling, solvent volatilization method or anti-solvent addition method;

and/or the organic solvent is one or more of alkane solvents, alcohol solvents, ketone solvents, ester solvents, aromatic hydrocarbon solvents, halogenated hydrocarbon solvents, nitrile solvents, ether solvents, aliphatic hydrocarbon solvents, N-dimethylformamide and dimethyl sulfoxide;

and/or 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 1:1 to 1:25 g/mL;

and/or the molar ratio of the fumaric acid to the 3- ((1H-pyrazolo [3,4-b ] pyridine-5-substituted) ethynyl) -4-methyl-N- (4- ((4-methylpiperazin-1-substituted) methyl) -3- (trifluoromethyl) phenyl) benzamide is 1:0.9 to 1: 3;

and/or, the temperature of the crystallization is 20-60 ℃;

and/or the crystallization time is 1-36h, more preferably 1-5 h;

and/or, when the crystallization adopts an anti-solvent addition method, the anti-solvent is an ester solvent.

6. The method according to claim 5, wherein the organic solvent is one or more of an alcohol solvent, a halogenated hydrocarbon solvent and an ether solvent;

and/or 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 1:5 to 1:25 g/mL;

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

and/or, when the crystallization adopts an anti-solvent addition method, the anti-solvent is ethyl acetate.

7. The method according to claim 6, wherein the organic solvent is one or more of methanol, ethanol, isopropanol, dichloromethane, and tetrahydrofuran.

8. A pharmaceutical composition comprising the crystalline form II of any one of claims 1-3, and a pharmaceutically acceptable excipient.

9. Use of the crystalline form II according to any one of claims 1 to 3, or the pharmaceutical composition according to claim 8, for the preparation of a medicament; the medicine is a medicine for preventing and/or treating 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.