HK1260782B - Crystalline form of gnrh receptor antagonist and preparation method therefor - Google Patents

Description

A crystalline form of a GnRH receptor antagonist and its preparation method

Technical Field

The present invention relates to a crystal form I of 1-(4-(7-(2,6-difluorobenzyl)-3-((dimethylamino)methyl)-5-(6-methoxypyridazin-3-yl)-4,6-dicarbonyl-4,5,6,7-tetrahydro-2H-pyrazolo[3,4-d]pyrimidin-2-yl)phenyl)-3-methoxyurea and a preparation method thereof, use thereof in a pharmaceutical composition, and use of the crystal form I and the composition in preparing a pharmaceutical composition for treating and/or preventing diseases associated with GnRH receptor antagonists.

Background Art

Endometriosis is a common estrogen-dependent gynecological disease that often occurs in women during their reproductive years. Its mechanism of action is still unclear. Currently, endometriosis is mainly diagnosed through laparoscopic surgery and treated surgically, or managed by taking birth control pills, GnRH receptor agonists, or progestins to reduce estrogen levels in the body.

Gonadotropin-releasing hormone (GnRH), also known as luteinizing hormone-releasing hormone (LHRH), is a central regulator of the endocrine reproductive system. The secretion and release of gonadotropins, such as luteinizing hormone (LH) and follicle-stimulating hormone (FSH), regulate the normal development of the ovaries and corpus luteum, playing a crucial role in the hypothalamic-pituitary-gonadal axis. GnRH receptors exert their regulatory effects by coupling with G proteins that activate the phosphatidylinositol calcium second messenger system. LH regulates the production of sex steroids, while FSH regulates spermatogenesis in men and follicle development in women.

LH and FSH are released into the circulation and bind to receptors on specific cells in the ovaries or testes, stimulating steroid production. In the presence of sex steroids, conditions such as endometriosis, uterine fibroids, and prostate cancer can worsen and require treatment with long-acting peptide GnRH receptor agonists and antagonists.

Peptide compounds present many challenges, including oral absorption, dosage form, dosage volume, drug stability, sustained action, and metabolic stability. The main reason small-molecule GnRH receptor antagonists are superior to existing peptide-based therapies is that they can be administered orally, making them convenient and quick.

The indirect tumor inhibition mechanism mediated by GnRH receptor agonists is through long-term action on the hypothalamus-pituitary-gonadal axis, leading to a decrease in pituitary gonadotropins (FSH, LH), thereby reducing the secretion of sex hormones and indirectly inhibiting the growth of tumor cells. GnRH receptor antagonists, on the other hand, directly inhibit the release of pituitary gonadotropins, thereby inhibiting the growth of tumor cells.

Currently, a series of patents for small molecule GnRH receptor antagonists have been disclosed, including WO2006096785, WO2010026993, WO2011076687, WO2012175514, etc. Small molecule GnRH receptor antagonists have good application prospects in the pharmaceutical industry as drugs. The applicant has provided a novel, highly effective, and low-toxic GnRH receptor antagonist in patent application WO2015062391A1 (published on May 7, 2015). The GnRH receptor antagonist has excellent effects and functions and can effectively treat endocrine and reproductive system diseases. Its chemical name is 1-(4-(7-(2,6-difluorobenzyl)-3-((dimethylamino)methyl)-5-(6-methoxypyridazin-3-yl)-4,6-dicarbonyl-4,5,6,7-tetrahydro-2H-pyrazolo[3,4-d]pyrimidin-2-yl)phenyl)-3-methoxyurea, and its structure is shown below.

The crystal structure of a pharmaceutically active ingredient often affects the chemical stability of the drug. Different crystal forms, preparation methods, and storage conditions may lead to changes in the crystal structure of the compound, sometimes accompanied by the production of other morphologies. Generally speaking, amorphous drug products do not have a regular crystal structure and often have other defects, such as poor product stability, difficulty in filtration, easy agglomeration, poor fluidity, etc. These differences often lead to difficulties in production scale-up. The stability of existing crystal forms needs to be improved. Therefore, it is necessary to improve the properties of the compound in all aspects. We need to conduct in-depth research to find new crystal forms with high crystal purity and good chemical stability.

Summary of the Invention

The technical problem to be solved by the present invention is to provide a crystalline form I of 1-(4-(7-(2,6-difluorobenzyl)-3-((dimethylamino)methyl)-5-(6-methoxypyridazin-3-yl)-4,6-dicarbonyl-4,5,6,7-tetrahydro-2H-pyrazolo[3,4-d]pyrimidin-2-yl)phenyl)-3-methoxyurea (as shown in formula (I)). The crystalline form has good stability, and the crystallization solvent used is low in toxicity and residue, and can be better applied in clinical practice.

The technical solutions of the present invention are as follows:

The present invention provides a crystal form I of a compound represented by formula (I), characterized in that: using Cu-Kα radiation, an X-ray powder diffraction pattern expressed in diffraction angle 2θ is obtained, and the crystal form I has characteristic peaks at 5.56, 9.15, 9.79, 11.08, 19.59, 20.25 and 22.16, wherein the error range of 2θ for each characteristic peak is ±0.2,

In a preferred embodiment of the present invention, the present invention provides a crystal form I of a compound represented by formula (I), characterized in that: using Cu-Kα radiation, an X-ray powder diffraction pattern expressed in diffraction angle 2θ is obtained, and the crystal form I has characteristic peaks at 5.56, 9.15, 9.79, 10.29, 11.08, 14.21, 16.61, 19.59, 20.25, 22.16 and 25.69, wherein the error range of each characteristic peak 2θ is ±0.2.

In a preferred embodiment of the present invention, the present invention provides a crystal form I of a compound represented by formula (I), characterized in that: using Cu-Kα radiation, an X-ray powder diffraction pattern expressed in diffraction angle 2θ is obtained, and the crystal form I has characteristic peaks at 5.22, 5.56, 9.15, 9.79, 10.29, 11.08, 13.38, 13.81, 14.21, 14.89, 16.61, 17.19, 18.47, 19.59, 20.25, 22.16, 23.32, 24.67, 25.69, 26.72, 28.73, 29.38, 31.78, 34.02 and 36.95, wherein the error range of each characteristic peak 2θ is ±0.2.

In a preferred embodiment of the present invention, the present invention provides a crystal form I of the compound represented by formula (I), characterized in that: the DSC melting endothermic peak is 160°C to 175°C, preferably 165°C to 170°C, and more preferably 168.17°C.

In a preferred embodiment of the present invention, the present invention further provides a method for preparing Form I of the compound represented by formula (I), the method comprising:

(1) Method 1, dissolving the compound represented by formula (I) in an organic solvent, crystallizing, filtering the crystals, washing, and drying to obtain the target crystal form I. The organic solvent is selected from alcohols, ketones, esters, ethers, a mixed solvent of ethers and alcohols, or a mixed solvent of ketones and water. The alcohol solvent is selected from methanol, ethanol, or isopropanol. The ketone solvent is selected from acetone. The ester solvent is selected from ethyl acetate. The ether solvent is selected from tetrahydrofuran. The mixed solvent of ethers and alcohols is selected from tetrahydrofuran/ethanol or tetrahydrofuran/isopropanol. The mixed solvent of ketones and water is selected from acetone/water. The ratio of the mixed solvent of the alcohol solvent to the ether solvent is 0.1:1 to 1:0.1, preferably tetrahydrofuran/ethanol = 1:1, tetrahydrofuran/isopropanol = 1:1. The ratio of the mixed solvent of the ketone to water is 0.1:1 to 1:0.1, preferably acetone/water = 5:1.

(2) Method 2: placing the compound represented by formula (I) in a solvent, beating the mixture, filtering the crystals, washing the crystals, and drying the mixture to obtain the target crystalline form I. The organic solvent is selected from alcohols, ketones, esters, ethers, a mixed solvent of ethers and alcohols, or a mixed solvent of ketones and water. The alcohol solvent is selected from methanol, ethanol, or isopropanol. The ketone solvent is selected from acetone. The ester solvent is selected from ethyl acetate. The ether solvent is selected from tetrahydrofuran. The mixed solvent of ethers and alcohols is selected from tetrahydrofuran/ethanol or tetrahydrofuran/ethanol. The mixed solvent of tetrahydrofuran/isopropanol, ketones and water is selected from acetone/water, the ratio of the mixed solvent of the alcohol solvent to the ether solvent is 0.1:1 to 1:0.1, preferably tetrahydrofuran/ethanol = 1:1, tetrahydrofuran/isopropanol = 1:1, the ratio of the mixed solvent of ketones and water is 0.1:1 to 1:0.1, preferably acetone/water = 1:1, the beating temperature is selected from room temperature to the boiling point of the solvent, the room temperature is preferably 15 to 30°C, more preferably 25°C.

The present invention further relates to a pharmaceutical composition of the crystalline form I of the compound represented by formula (I), characterized in that it contains one or more pharmaceutically acceptable carriers, diluents or excipients.

The present invention further relates to the use of the crystal form I of the compound represented by formula (I) or a pharmaceutical composition of the crystal form I in the preparation of a drug for treating and/or preventing diseases associated with GnRH receptor antagonists, wherein the diseases are selected from endocrine reproductive system diseases.

The structure and crystal form of the obtained crystal form of the compound represented by formula (I) were determined by X-ray powder diffraction (XRPD) and differential scanning calorimetry (DSC), and the residual solvent in the obtained crystals was detected.

The crystalline form of compound I represented by formula (I) prepared according to the method of the present invention contains no or only a low content of residual solvents, which meets the limit requirements for residual solvents in pharmaceutical products specified in the National Pharmacopoeia. Therefore, the crystals of the present invention can be preferably used as pharmaceutical active ingredients.

The recrystallization method is not particularly limited and can be carried out using conventional recrystallization methods. For example, the compound represented by formula (I) can be dissolved in an organic solvent by heating and then slowly cooled to crystallize. After crystallization is complete, the desired crystals can be obtained by filtration and drying.

The crystallization methods of the present invention include room temperature crystallization, cooling crystallization, etc.

The starting material used in the method for preparing the crystal form of the present invention can be any form of the compound represented by formula (I), including but not limited to: amorphous form, any crystal form, etc.

Detailed Description of the Invention

In the specification and claims of the application, unless otherwise indicated, the scientific and technological terms used herein have the implication commonly understood by those skilled in the art. However, in order to better understand the present invention, the definition and explanation of some related terms are provided below. In addition, when the definition and explanation of the term provided in the application are inconsistent with the implication commonly understood by those skilled in the art, the definition and explanation of the term provided in the application shall be as the criterion.

The "beating" mentioned in the present invention refers to a purification method that utilizes the property that a substance has poor solubility in a solvent but impurities have good solubility in a solvent. Beating purification can remove color, change the crystal form or remove a small amount of impurities.

The "C _1-6 alkyl" mentioned in the present invention refers to a straight-chain or branched alkyl group containing 1 to 6 carbon atoms. Specific examples include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, isopentyl, 2-methylbutyl, neopentyl, 1-ethylpropyl, n-hexyl, isohexyl, 3-methylpentyl, 2-methylpentyl, 1-methylpentyl, 3,3-dimethylbutyl, 2,2-dimethylbutyl, 1,1-dimethylbutyl, 1,2-dimethylbutyl, 1,3-dimethylbutyl, 2,3-dimethylbutyl, 2-ethylbutyl, 1,2-dimethylpropyl, and the like.

The "hydroxyl group" mentioned in the present invention refers to groups such as -OH.

The "ketone solvent" described in the present invention refers to a compound in which a carbonyl group (-C(O)-) is connected to two hydrocarbon groups. Depending on the different hydrocarbon groups in the molecule, ketones can be divided into aliphatic ketones, alicyclic ketones, aromatic ketones, saturated ketones and unsaturated ketones. Specific examples include but are not limited to: acetone, methyl butyl ketone or methyl isobutyl ketone.

The "ester solvent" mentioned in the present invention refers to a combination of a lower organic acid containing 1 to 4 carbon atoms and a lower alcohol containing 1 to 6 carbon atoms. Specific examples include but are not limited to ethyl acetate, isopropyl acetate or butyl acetate.

The "ether solvent" described in the present invention refers to a chain compound or a cyclic compound containing an ether bond -O- and having 1 to 10 carbon atoms. Specific examples include but are not limited to propylene glycol methyl ether, tetrahydrofuran or 1,4-dioxane.

The "alcohol solvent" described in the present invention refers to a group derived from one or more "hydroxyl groups" replacing one or more hydrogen atoms on a "C _1-6 alkyl group". The "hydroxyl group" and "C _1-6 alkyl group" are as defined above. Specific examples include but are not limited to: methanol, ethanol, propanol or 2-propanol.

The "mixed solvent" described in the present invention refers to a solvent formed by mixing one or more different types of organic solvents in a certain proportion, or a solvent formed by mixing an organic solvent and water in a certain proportion, wherein the proportion is a volume ratio, and the volume ratio is selected from 0.1:1 to 1:0.1, preferably 1:1 or 5:1; the mixed solvent is preferably a mixed solvent of alcohols and ethers, a mixed solvent of alcohol solvents and water, or a mixed solvent of ketone solvents and water.

The "X-ray powder diffraction pattern or XRPD" mentioned in the present invention refers to the Bragg equation 2d sin θ = nλ (wherein λ is the wavelength of the X-rays, and the diffraction order n is any positive integer, generally the first-order diffraction peak is taken, n = 1). When X-rays are incident at a grazing angle θ (the complementary angle of the incident angle, also known as the Bragg angle) on an atomic plane with a lattice plane spacing d in a crystal or a portion of a crystalline sample, the Bragg equation is satisfied, thereby obtaining this set of X-ray powder diffraction patterns.

The "differential scanning calorimetry or DSC" mentioned in the present invention refers to measuring the temperature difference and heat flow difference between a sample and a reference object during the process of heating or maintaining the sample at a constant temperature, so as to characterize all physical and chemical changes related to thermal effects and obtain phase change information of the sample.

The "2θ or 2θ angle" mentioned in the present invention refers to the diffraction angle, θ is the Bragg angle, the unit is ° or degree, and the error range of 2θ is ±0.1 to ±0.5, preferably ±0.1 to ±0.3, and more preferably ±0.2.

The term "interplanar spacing" (d-value) used in this invention refers to the three non-parallel unit vectors a, b, and c connecting two adjacent lattice points in a spatial lattice. These vectors divide the lattice into juxtaposed parallelepiped units, known as the interplanar spacing. The spatial lattice is divided along the lines connecting the parallelepiped units to form a set of rectilinear grids, known as spatial lattices or crystal lattices. The lattice and crystal lattice, respectively, use geometric points and lines to reflect the periodicity of the crystal structure. Different crystal planes have different interplanar spacings (i.e., the distance between two adjacent parallel crystal planes); the units are Å or Å.

The present invention also relates to a pharmaceutical composition comprising Form I of the compound represented by Formula (I), and optionally one or more pharmaceutical carriers and/or diluents. The pharmaceutical composition can be prepared in any pharmaceutically acceptable dosage form. For example, Form I or the pharmaceutical preparation of the present invention can be formulated as a tablet, capsule, pill, granule, solution, suspension, syrup, injection (including injection, sterile powder for injection and concentrated solution for injection), suppository, inhalant or spray.

Furthermore, the pharmaceutical compositions of the present invention can be administered to patients or subjects in need of such treatment by any suitable route of administration, such as oral, parenteral, rectal, pulmonary, or topical administration. For oral administration, the pharmaceutical compositions can be formulated into oral preparations, such as solid oral preparations such as tablets, capsules, pills, granules, or oral liquid preparations such as oral solutions, oral suspensions, and syrups. When formulated into oral preparations, the pharmaceutical preparations can also contain suitable fillers, binders, disintegrants, lubricants, and the like. For parenteral administration, the pharmaceutical preparations can be formulated into injectable preparations, including injectable solutions, sterile powders for injection, and concentrated solutions for injection. When formulated into injectable preparations, the pharmaceutical compositions can be produced using conventional methods in the pharmaceutical industry. When formulated into injectable preparations, additives may be omitted or added as appropriate, depending on the properties of the drug. For rectal administration, the pharmaceutical preparations can be formulated into suppositories, etc. For pulmonary administration, the pharmaceutical preparations can be formulated into inhalers or sprays, etc. In certain preferred embodiments, the crystalline form I of the present invention is present in a pharmaceutical composition or drug in a therapeutically and/or prophylactically effective amount. In certain preferred embodiments, the crystalline form I of the present invention is present in a pharmaceutical composition or drug in the form of a unit dose.

The crystal form I of the compound of formula (I) of the present invention can be used to prepare a method for treating and/or preventing diseases associated with GnRH receptor antagonists. Therefore, the present application also relates to the use of the crystal form I of the compound of formula (I) of the present invention for preparing a medicament, which is used to treat and/or prevent diseases associated with GnRH receptor antagonists in a subject. In addition, the present application also relates to a method for inhibiting a disease associated with a GnRH receptor antagonist, which comprises administering a therapeutically and/or prophylactically effective amount of the crystal form I of the compound of formula (I) of the present invention, or a pharmaceutical composition of the present invention, to a subject in need thereof.

In certain preferred embodiments, the disease is a disease associated with a GnRH receptor antagonist selected from the group consisting of endocrine reproductive system diseases.

Advantageous Effects of the Invention

Compared with the prior art, the technical solution of the present invention has the following advantages:

(1) The crystal form I of the compound represented by formula (I) of the present invention contains no or only a low content of residual solvents, which meets the limit requirements for residual solvents in pharmaceutical products stipulated in the National Pharmacopoeia. Therefore, the crystal of the present invention can be preferably used as a pharmaceutical active ingredient.

(2) Studies have shown that the crystal form I of the compound represented by formula (I) prepared by the present invention has a high purity. The crystal form does not change under conditions of light, high temperature, and high humidity by XRPD detection, and the crystal form has good stability; the HPLC purity changes little and the chemical stability is high; the crystal form I of the compound represented by formula (I) obtained by the technical solution of the present invention can meet the pharmaceutical requirements of production, transportation, and storage, and the production process is stable, repeatable, and controllable, and can be adapted to industrial production.

BRIEF DESCRIPTION OF THE DRAWINGS

Figure 1 is the XRPD pattern of the crystalline form of compound I represented by formula (I).

Figure 2 is the DSC spectrum of the crystal form of compound I represented by formula (I).

Figure 3 is the XRPD pattern of the crystalline form of compound A represented by formula (I).

FIG4 is a DSC spectrum of the crystalline form of compound A represented by formula (I).

Figure 5 is the XRPD pattern of the crystalline form of compound B represented by formula (I).

FIG6 is a DSC spectrum of the crystalline form of compound B represented by formula (I).

Figure 7 is the XRPD pattern of the crystalline form of compound C represented by formula (I).

FIG8 is a DSC spectrum of the crystalline form of compound C represented by formula (I).

FIG9 is an XRPD pattern of the crystalline form of compound D represented by formula (I).

FIG10 is a DSC spectrum of the crystalline form of compound D represented by formula (I).

DETAILED DESCRIPTION

The present invention will be explained in more detail below with reference to the embodiments. The embodiments of the present invention are only used to illustrate the technical solutions of the present invention and are not intended to limit the essence and scope of the present invention.

Test conditions of the instruments used in the experiment:

1. Differential Scanning Calorimeter (DSC)

Instrument model: Mettler Toledo DSC 1 STAR ^e System

Purge gas: nitrogen

Heating rate: 10.0℃/min

Temperature range: 40-300℃

2. X-ray Powder Diffraction (XRPD)

Instrument model: Bruker D8Focus X-ray powder diffractometer

Radiation: Monochromatic Cu-Kα radiation (λ=1.5406)

Scanning mode: θ/2θ, scanning range: 2-40°

Voltage: 40kV, Current: 40mA

Example 1

The crude product of 1-(4-(7-(2,6-difluorobenzyl)-3-((dimethylamino)methyl)-5-(6-methoxypyridazin-3-yl)-4,6-dicarbonyl-4,5,6,7-tetrahydro-2H-pyrazolo[3,4-d]pyrimidin-2-yl)phenyl)-3-methoxyurea (300 mg, 0.49 mmol) (prepared according to the method of Example 11 of WO2015062391A1) was added to the reaction flask, and a mixed solvent of acetone/water (5 mL, V:V=5:1) was added. The mixture was heated to reflux until all the solids dissolved. The heating was stopped, the mixture was cooled to crystallize, filtered, and dried in vacuo to obtain 212 mg of a solid. The diffraction angle 2θ of the crystalline sample detected by XRPD was 5.19 (17.02), 5.48 (16.10), 9.08 (9.73), 9.73 (9.08), 10.24 (8.63), 11.01 (8.03), 13.80 (6.41), 14.13 (6.26), 14.82 (5.97), 15.35 (5.77), 16.56 (5.35), 18.31 (4.84), There are characteristic peaks at 18.65 (4.75), 19.50 (4.55), 20.18 (4.40), 22.07 (4.03), 23.26 (3.82), 24.59 (3.62), 25.61 (3.48), 26.66 (3.34), 28.69 (3.11), 29.30 (3.05), 33.96 (2.64) and 36.91 (2.43), and this crystal form is defined as crystal form I.

Example 2

The crude compound of formula (I) (300 mg, 0.49 mmol) (prepared according to the method of Example 11 of WO2015062391A1) was added to a reaction flask, acetone (6 mL) was added, and the mixture was stirred at room temperature overnight. The mixture was filtered and dried under vacuum to obtain 221 mg of a solid. The XRPD pattern of the crystalline sample is shown in Figure 1, and its DSC spectrum is shown in Figure 2. The DSC melting endotherm peak is around 168.17°C, the onset melting temperature is 154.23°C, and the characteristic peak positions are shown in the following table:

Table 1. Characteristic peaks of Form I

Example 3

The crude compound of formula (I) (500 mg, 0.82 mmol) (prepared according to Example 11 of WO2015062391A1) was added to a reaction flask, and methanol (50 mL) was added. The mixture was heated to reflux. After the solid dissolved, heating was stopped, and the mixture was stirred for crystallization. The mixture was filtered and vacuum dried to obtain 350 mg of a solid. The XRPD and DSC spectra of the crystalline sample were compared and confirmed to be Form I.

Example 4

The crude compound of formula (I) (500 mg, 0.82 mmol) (prepared according to Example 11 of WO2015062391A1) was added to a reaction flask, and ethanol (125 mL) was added. The mixture was heated to reflux. After the solid dissolved, heating was stopped, and the mixture was stirred for crystallization. The mixture was filtered and dried under vacuum to obtain 406 mg of a solid. The XRPD and DSC spectra of the crystalline sample were compared and confirmed to be Form I.

Example 5

The crude compound of formula (I) (500 mg, 0.82 mmol) (prepared according to Example 11 of WO2015062391A1) was added to a reaction flask, and isopropanol (10 mL) was added. The mixture was stirred at room temperature overnight, filtered, and dried under vacuum to obtain 445 mg of a solid. The XRPD and DSC spectra of the crystalline sample were compared and confirmed to be Form I.

Example 6

The crude compound of formula (I) (500 mg, 0.82 mmol) (prepared according to Example 11 of WO2015062391A1) was added to a reaction flask, and acetone (25 mL) was added. The mixture was heated to reflux. After the solid dissolved, heating was stopped, and the mixture was stirred for crystallization. The mixture was filtered and vacuum dried to obtain 251 mg of a solid. The XRPD and DSC spectra of the crystalline sample were compared and confirmed to be Form I.

Example 7

The crude compound of formula (I) (300 mg, 0.49 mmol) (prepared according to Example 11 of WO2015062391A1) was added to a reaction flask, and ethyl acetate (9 mL) was added. The mixture was stirred at room temperature overnight, filtered, and dried under vacuum to obtain 224 mg of a solid. The XRPD and DSC spectra of the crystalline sample were compared and confirmed to be Form I.

Example 8

The crude compound of formula (I) (300 mg, 0.49 mmol) (prepared according to Example 11 of WO2015062391A1) was added to a reaction flask. Tetrahydrofuran/ethanol (8 mL, V:V = 1:1) was added and heated to reflux. After the solid dissolved, heating was stopped. The mixture was stirred to crystallize, filtered, and vacuum dried to obtain 197 mg of a solid. Comparison of the XRPD and DSC spectra of the crystalline sample confirmed that the product was Form I.

Example 9

The crude compound of formula (I) (300 mg, 0.49 mmol) (prepared according to Example 11 of WO2015062391A1) was added to a reaction flask. Tetrahydrofuran/isopropanol (12 mL, V:V = 1:1) was added and heated to reflux. After the solid dissolved, heating was stopped. The mixture was stirred to crystallize, filtered, and vacuum dried to obtain 182 mg of a solid. Comparison of the XRPD and DSC spectra of the crystalline sample confirmed that the product was Form I.

Example 10

The crude compound of formula (I) (300 mg, 0.49 mmol) (prepared according to Example 11 of WO2015062391A1) was added to a reaction flask, followed by methanol (6 mL). The mixture was stirred overnight at room temperature, filtered, and dried under vacuum to yield 239 mg of a solid. Comparison of the XRPD and DSC spectra of the crystalline sample confirmed that the product was Form I.

Example 11

The crude compound of formula (I) (300 mg, 0.49 mmol) (prepared according to Example 11 of WO2015062391A1) was added to a reaction flask, followed by ethanol (6 mL). The mixture was stirred overnight at room temperature, filtered, and dried under vacuum to yield 231 mg of a solid. Comparison of the XRPD and DSC spectra of the crystalline sample confirmed that the product was Form I.

Example 12

1-(4-(7-(2,6-difluorobenzyl)-3-((dimethylamino)methyl)-5-(6-methoxypyridazin-3-yl)-4,6-dicarbonyl-4,5,6,7-tetrahydro-2H-pyrazolo[3,4-d]pyrimidin-2-yl)phenyl)-3-methoxyurea (500 mg, 0.82 mmol) (prepared according to Example 1) was added to a reaction flask, and purified water (10 mL) was added. The mixture was slurried at room temperature for 5 hours, filtered, and dried to obtain 369 mg of a solid. XRPD and DSC analysis confirmed that the crystalline form was Form A.

The obtained Form A was added to a reaction flask, ethanol (4 mL) was added, stirred at room temperature overnight, filtered, and dried in vacuo to obtain 89 mg of a solid. The XRPD and DSC spectra of the crystalline sample were compared and confirmed to be Form I.

Example 13

1-(4-(7-(2,6-difluorobenzyl)-3-((dimethylamino)methyl)-5-(6-methoxypyridazin-3-yl)-4,6-dicarbonyl-4,5,6,7-tetrahydro-2H-pyrazolo[3,4-d]pyrimidin-2-yl)phenyl)-3-methoxyurea (300 mg, 0.49 mmol) (prepared according to Example 1) was added to a reaction flask, and acetonitrile (9 mL) was added. The mixture was heated to reflux until all the solid dissolved. Heating was stopped, and the mixture was cooled to crystallize. The mixture was filtered and dried to obtain 243 mg of a solid. XRPD and DSC analysis confirmed that the crystalline form was Form B.

The obtained Form B was added to a reaction flask, ethanol (4 mL) was added, stirred at room temperature overnight, filtered, and dried in vacuo to obtain 80 mg of a solid. The XRPD and DSC spectra of the crystalline sample were compared and confirmed to be Form I.

Example 14

1-(4-(7-(2,6-difluorobenzyl)-3-((dimethylamino)methyl)-5-(6-methoxypyridazin-3-yl)-4,6-dicarbonyl-4,5,6,7-tetrahydro-2H-pyrazolo[3,4-d]pyrimidin-2-yl)phenyl)-3-methoxyurea (300 mg, 0.49 mmol) (prepared according to Example 1) was added to a reaction flask, and 1,4-dioxane (15 mL) was added. The mixture was heated to reflux until all the solid dissolved. Heating was stopped, and the mixture was cooled to crystallize. The mixture was filtered and dried to obtain 205 mg of a solid. XRPD and DSC analysis confirmed that the crystalline form was Form C.

The resulting Form C was added to a reaction flask, ethanol (10 mL) was added, and the mixture was stirred at room temperature overnight. The mixture was filtered and dried under vacuum to obtain 78 mg of a solid with a yield of 78.0%. The XRPD and DSC patterns of the crystalline sample were compared to confirm that the product was Form I.

Example 15

1-(4-(7-(2,6-difluorobenzyl)-3-((dimethylamino)methyl)-5-(6-methoxypyridazin-3-yl)-4,6-dicarbonyl-4,5,6,7-tetrahydro-2H-pyrazolo[3,4-d]pyrimidin-2-yl)phenyl)-3-methoxyurea (300 mg, 0.49 mmol) (prepared according to Example 1) was added to a reaction flask, and tetrahydrofuran/water (15 mL, V:V=1:1) was added. The mixture was heated to reflux until all the solid dissolved. Heating was stopped, and the mixture was cooled to crystallize, filtered, and dried to obtain 205 mg of a solid. XRPD and DSC analysis confirmed that the crystalline form was Form D.

The obtained Form D was added to a reaction flask, ethanol (10 mL) was added, stirred at room temperature overnight, filtered, and dried in vacuo to obtain 78 mg of a solid. The XRPD and DSC spectra of the crystalline sample were compared and confirmed to be Form I.

Example 16

The Form I product sample obtained in Example 1 and the Form A, B, C, and D product samples obtained in Examples 12, 13, 14, and 15 were laid out open to examine the stability of the samples under conditions of illumination (4500 Lux), heating (40° C., 60° C.), and high humidity (RH75%, RH90%). The sampling time was 5 days and 10 days, and the purity results of the HPLC test are shown in Table 2.

Test results:

Table 2. Comparison of the stability of the compound of formula (I) of the present invention in crystal form I and crystal forms A, B, C, and D

Test conclusion

The stability test results in Table 2 show that:

Under conditions of light, high humidity, and high temperature exposure, the HPLC purity data of Form I of the compound represented by formula (I) decreased less than that of Forms A, B, C, and D, and XRPD detection showed no change in the form, indicating that the stability of Form I of the present invention is significantly better than that of Forms A, B, C, and D.

Example 17

The crystalline form sample of Compound I represented by Formula (I) obtained by the method of Example 1 was ground, heated and tableted. The crystal stability, XRPD and DSC test results of the sample are shown in Table 3.

Test results:

Table 3. Study on the special stability of the crystal form of compound I represented by formula (I)

Test conclusion:

The stability test data in Table 3 show that the crystal form does not change during the grinding, heating and tableting processes, indicating that the crystal form I of the present invention has high stability.

Claims (8)

1. The I crystal form of the compound shown in formula (I), characterized in that: an X-ray powder diffraction pattern, expressed as a diffraction angle 2θ, is obtained using Cu-Kα radiation, wherein the I crystal form has characteristic peaks at 5.56, 9.15, 9.79, 11.08, 19.59, 20.25, and 22.16, wherein the error range of 2θ for each characteristic peak is ±0.2. 2. The I-type according to claim 1, characterized in that the I-type has characteristic peaks at 5.56, 9.15, 9.79, 10.29, 11.08, 14.21, 16.61, 19.59, 20.25, 22.16 and 25.69, wherein the error range of 2θ for each characteristic peak is ±0.2. 3. The I-type according to claim 2, characterized in that the I-type has characteristic peaks at 5.22, 5.56, 9.15, 9.79, 10.29, 11.08, 13.38, 13.81, 14.21, 14.89, 16.61, 17.19, 18.47, 19.59, 20.25, 22.16, 23.32, 24.67, 25.69, 26.72, 28.73, 29.38, 31.78, 34.02, and 36.95, wherein the error range of 2θ for each characteristic peak is ±0.2. 4. A method for preparing the I crystal form of the compound of formula (I) as described in any one of claims 1-3, characterized in that the method comprises: 1) Method 1: Dissolve the compound shown in formula (I) in an organic solvent, crystallize, filter the crystals and wash, and dry to obtain the target crystal form I. The organic solvent is selected from alcohols, ketones, esters, ethers, mixed solvents of ethers and alcohols or mixed solvents of ketones and water. 2) Method 2: Place the compound shown in formula (I) in a solvent, slurry it, filter and crystallize it, wash it, and dry it to obtain the target crystal form I. The organic solvent is selected from alcohols, ketones, esters, ethers, mixed solvents of ethers and alcohols, or mixed solvents of ketones and water. Furthermore, in Method 1 and Method 2, the alcohol solvent is selected from methanol, ethanol or isopropanol, the ketone solvent is acetone, the ester solvent is ethyl acetate, the ether solvent is tetrahydrofuran, the mixed solvent of ether and alcohol is selected from tetrahydrofuran/ethanol or tetrahydrofuran/isopropanol, and the mixed solvent of ketone and water is acetone/water. 5. A crystal form I as described in any one of claims 1-3, characterized in that: the melting endothermic peak of the DSC is 160℃～175℃. 6. The I-type crystal according to claim 5, characterized in that: the melting endothermic peak of the DSC is 165℃～170℃. 7. The I-type crystal according to claim 6, characterized in that: the melting endothermic peak of the DSC is 168.17℃. 8. A pharmaceutical composition comprising the I crystal form according to any one of claims 1-3, characterized in that it comprises one or more pharmaceutically acceptable carriers.