CN115073430A

CN115073430A - Crystal form of Belumosudil mesylate and preparation method and application thereof

Info

Publication number: CN115073430A
Application number: CN202210191092.0A
Authority: CN
Inventors: 陈敏华; 施文睿; 张婧
Original assignee: Crystal Pharmaceutical Suzhou Co Ltd
Current assignee: Crystal Pharmaceutical Suzhou Co Ltd
Priority date: 2021-03-10
Filing date: 2022-02-28
Publication date: 2022-09-20

Abstract

The invention relates to a crystal form of Belumosudil (hereinafter referred to as compound I) mesylate, a preparation method thereof, a pharmaceutical composition containing the crystal form, and application of the crystal form in preparing ROCK2 inhibitor drugs and drugs for treating chronic graft-versus-host disease, systemic sclerosis and idiopathic pulmonary fibrosis. Compared with the prior art, the crystal form of the Belumosudil mesylate provided by the inventionThe method has one or more improved properties, and has important value for optimizing and developing the medicine in the future.

Description

Crystal form of Belumosudil mesylate and preparation method and application thereof

Technical Field

The present invention relates to the field of crystal chemistry. In particular to a crystal form of Belumosudil mesylate, a preparation method and application thereof.

Background

Chronic graft versus host disease (cGVHD) is an immune-mediated inflammatory and fibrotic disease. It is a major cause of morbidity, mortality, and impaired quality of life (QOL) following allogeneic hematopoietic cell transplantation (alloHCT). cGVHD affects up to 70% of alloHCT recipients. The incidence of cGVHD is 20% -50% in children that survive more than 100 days after alloHCT. It is the leading cause of non-recurrent death for more than 2 years post-surgery in alloHCT.

REZUROCK ^TM (Belumosudil) is approved in the united states for the treatment of adult and pediatric patients 12 years old and older with cGVHD after failure of at least two previous systemic treatment regimens. Belumosudil is the only approved therapy targeting Rho-associated coiled coil protein kinase 2(ROCK 2). Research into the use of Belumosudil for the treatment of systemic sclerosis is under development and the FDA has granted Belumosudil an orphan drug for the treatment of systemic sclerosis.

The chemical name of Belumosudil is 2- (3- (4- (1H-indazol-5-ylamino) quinazolin-2-yl) phenoxy) -N-isopropylacetamide (hereinafter "Compound I"), the structural formula of which is as follows:

a crystal is a solid in which molecules of a compound are three-dimensionally ordered in a microstructure to form a lattice. Polymorphism refers to the phenomenon of multiple crystalline forms of a compound. The compounds may exist in one or more crystalline forms, but their presence and identity are not specifically contemplated. The raw material medicines with different crystal forms have different physicochemical properties, which may cause different dissolution and absorption of the medicine in vivo, thereby affecting the clinical curative effect of the medicine to a certain extent. In particular, the crystal form of some insoluble oral solid or semisolid preparations is crucial to the product performance. In addition, the physicochemical properties of the crystal form are crucial to the production process. Thus, polymorphism is an important part of drug research and drug quality control.

The prior art does not disclose solid forms of compound I mesylate, only the preparation of compound I and the brown solid of compound I. Among them, WO2006105081a2 discloses a method for preparing compound I: the crude compound I was purified using preparative HPLC to give compound I in turn, without disclosing the form of compound I. CN106916145B discloses a brown solid of compound I. The inventor of the application repeats the preparation methods disclosed in WO2006105081A2 and CN106916145B to obtain the solid of the compound I, which is named as the prior art P1 and the prior art P2 respectively, and researches the solid, and finds that the prior art has the problems of high solvent content, low solubility, high hygroscopicity, poor stability and the like, and is not suitable for medicinal development.

In order to overcome the defects of the prior art, the inventors of the present application have conducted intensive studies on compound I and salts thereof, and have unexpectedly found that the crystal form of compound I mesylate provided by the present invention has advantages in at least one aspect of solubility, hygroscopicity, purification effect, stability, adhesiveness, compressibility, fluidity, in vitro and in vivo dissolution, bioavailability, and the like, and particularly has no solvent residue, high solubility, good stability, and low hygroscopicity, thereby solving the problems of the prior art, and having very important significance in the development of drugs containing compound I.

Disclosure of Invention

The invention mainly aims to provide a crystal form of compound I mesylate, a preparation method thereof and a pharmaceutical composition containing the crystal form.

According to an object of the present invention, the present invention provides a mesylate salt form CSII of compound I (hereinafter referred to as "form CSII").

On one hand, the X-ray powder diffraction pattern of the crystal form CSII has characteristic peaks at diffraction angle 2 theta values of 6.3 degrees +/-0.2 degrees, 12.7 degrees +/-0.2 degrees and 15.9 degrees +/-0.2 degrees by using Cu-Kalpha radiation.

Further, using Cu-K α radiation, the X-ray powder diffraction pattern of crystalline form CSII has characteristic peaks at 1, or 2, or 3 of diffraction angle 2 θ values of 7.9 ° ± 0.2 °, 19.2 ° ± 0.2 °, 19.9 ° ± 0.2 °; preferably, the X-ray powder diffraction pattern of the crystal form CSII has a characteristic peak at 3 points with diffraction angle 2 theta values of 7.9 +/-0.2 degrees, 19.2 +/-0.2 degrees and 19.9 +/-0.2 degrees.

Further, using Cu-K α radiation, the X-ray powder diffraction pattern of crystalline form CSII has a characteristic peak at 1, or 2, of diffraction angle 2 θ values of 14.5 ° ± 0.2 °, 20.4 ° ± 0.2 °; preferably, the X-ray powder diffraction spectrum of the crystal form CSII has a characteristic peak at 2 of diffraction angle 2 theta values of 14.5 degrees +/-0.2 degrees and 20.4 degrees +/-0.2 degrees.

In another aspect, the crystalline form CSII has an X-ray powder diffraction pattern with characteristic peaks at any 1, or 2, or 3, or 4, or 5, or 6, or 7, or 8, or 9, or 10, or 11 of diffraction angle 2 Θ values of 6.3 ° ± 0.2 °, 12.7 ° ± 0.2 °, 15.9 ° ± 0.2 °,7.9 ° ± 0.2 °, 19.2 ° ± 0.2 °, 19.9 ° ± 0.2 °, 14.5 ° ± 0.2 °, 20.4 ° ± 0.2 °,9.9 ° ± 0.2 °, 25.2 ° ± 0.2 °, 26.5 ° ± 0.2 °, using Cu-K α radiation.

Without limitation, the X-ray powder diffraction pattern of crystalline form CSII is substantially as shown in figure 3.

Without limitation, crystalline form CSII has a mass loss of about 5.9% when heated to 100 ℃, with a thermogravimetric analysis profile substantially as shown in figure 4.

Without limitation, the crystalline form CSII has an endothermic peak at about 100 ℃, an exothermic peak at about 206 ℃ and an endothermic peak at about 255 ℃ (about 253 ℃ C.), and a Differential Scanning Calorimetry (DSC) chart is shown in FIG. 5.

Without limitation, crystalline form CSII is a hydrate.

Without limitation, the crystalline form CSII contains no more than 10% by mass of water.

Further, the crystalline form CSII contains not more than 8% by mass of water.

Further, the crystalline form CSII contains not more than 6% by mass of water.

According to an object of the present invention, the present invention also provides a preparation method of the crystalline form CSII, the preparation method comprising:

and mixing the solid of the compound I, a mixed solvent of methanesulfonic acid, alcohols and water or a mixed solvent of ethers and water, and stirring to obtain the crystal form CSII.

Further, the molar ratio of the compound I solid to methanesulfonic acid is preferably 0.9:1 to 1.1:1, more preferably 1: 1; the alcohol solvent is preferably isopropanol; the ether solvent is preferably tetrahydrofuran; the volume ratio of the alcohol to the water or the ether to the water in the mixed solvent is preferably 9: 1; optionally, the resulting CSII is dried at less than 45 ℃.

According to an object of the present invention, the present invention provides a mesylate salt form CSIII of compound I (hereinafter referred to as "form CSIII").

On the one hand, the X-ray powder diffraction pattern of the crystal form CSIII has characteristic peaks at diffraction angle 2 theta values of 7.3 degrees +/-0.2 degrees, 16.6 degrees +/-0.2 degrees and 19.0 degrees +/-0.2 degrees by using Cu-Kalpha radiation.

Further, using Cu-K α radiation, the X-ray powder diffraction pattern of crystalline form CSIII has characteristic peaks at 1, or 2, or 3 of diffraction angle 2 θ values of 6.9 ° ± 0.2 °, 17.4 ° ± 0.2 °, 19.6 ° ± 0.2 °; preferably, the X-ray powder diffraction pattern of the crystal form CSIII has a characteristic peak at 3 points with diffraction angle 2 theta values of 6.9 +/-0.2 degrees, 17.4 +/-0.2 degrees and 19.6 +/-0.2 degrees.

Further, using Cu-K α radiation, the X-ray powder diffraction pattern of crystalline form CSIII has characteristic peaks at 1, or 2, or 3 of diffraction angle 2 θ values of 12.8 ° ± 0.2 °, 14.6 ° ± 0.2 °, 25.9 ± 0.2 °; preferably, the X-ray powder diffraction spectrum of the crystal form CSIII has a characteristic peak at 3 points with diffraction angle 2 theta values of 12.8 degrees +/-0.2 degrees, 14.6 degrees +/-0.2 degrees and 25.9 degrees +/-0.2 degrees.

In another aspect, the crystalline form CSIII has an X-ray powder diffraction pattern with characteristic peaks at diffraction angle 2 θ values of 7.3 ° ± 0.2 °, 16.6 ° ± 0.2 °, 19.0 ° ± 0.2 °, 6.9 ° ± 0.2 °, 17.4 ° ± 0.2 °, 19.6 ° ± 0.2 °, 12.8 ° ± 0.2 °, 14.6 ° ± 0.2 °, 25.9 ± 0.2 °, 9.4 ± 0.2 °, 13.7 ± 0.2 °, and any 1, or 2, or 3, or 4, or 5, or 6, or 7, or 8, or 9, or 10, or 11, or 12 using Cu-ka radiation.

Without limitation, the X-ray powder diffraction pattern of crystalline form CSIII is substantially as shown in figure 12.

Without limitation, crystalline form CSIII has a mass loss of about 0.1% when heated to 200 ℃, with a thermogravimetric analysis profile substantially as shown in figure 13.

Without limitation, crystalline form CSIII has an endothermic peak at about 253 ℃ (onset temperature is about 252 ℃), and a differential scanning calorimetry thermogram is shown in FIG. 14.

Without limitation, crystalline form CSIII is an anhydrate.

According to an object of the present invention, the present invention also provides a preparation method of the crystalline form CSIII, the preparation method comprising:

mixing the solid of the compound I, methanesulfonic acid and an ether/water mixed solvent, stirring, separating to obtain a solid, and heating the solid to more than 190 ℃ to obtain the crystal form CSIII.

Further, the molar charge ratio of the compound I solid to methanesulfonic acid is preferably 0.9:1 to 1.1:1, more preferably 1: 1; the ether solvent is preferably tetrahydrofuran; the volume ratio of the ether solvent to water is preferably 9: 1; the heating temperature is preferably 190 ℃ to 235 ℃.

According to an object of the present invention, the present invention provides a crystalline form CSIV of the mesylate salt of compound I (hereinafter referred to as "crystalline form CSIV").

On the one hand, the X-ray powder diffraction pattern of the crystal form CSIV has characteristic peaks at diffraction angle 2 theta values of 6.5 degrees +/-0.2 degrees, 16.3 degrees +/-0.2 degrees and 19.8 degrees +/-0.2 degrees by using Cu-Kalpha radiation.

Further, using Cu-K α radiation, the X-ray powder diffraction pattern of crystalline form CSIV has characteristic peaks at 1, or 2, or 3 of diffraction angle 2 θ values of 8.1 ° ± 0.2 °, 12.9 ° ± 0.2 °, 20.5 ° ± 0.2 °; preferably, the X-ray powder diffraction pattern of the crystal form CSIV has a characteristic peak at 3 points with diffraction angles 2 theta of 8.1 +/-0.2 degrees, 12.9 +/-0.2 degrees and 20.5 +/-0.2 degrees.

Further, using Cu-Ka radiation, the X-ray powder diffraction pattern of the crystal form CSIV has characteristic peaks at 1 or 2 of diffraction angle 2 theta values of 10.2 degrees +/-0.2 degrees and 19.5 degrees +/-0.2 degrees; preferably, the X-ray powder diffraction pattern of the crystal form CSIV has characteristic peaks at 2 positions with diffraction angles 2 theta of 10.2 degrees +/-0.2 degrees and 19.5 degrees +/-0.2 degrees.

In another aspect, the crystalline form CSIV has an X-ray powder diffraction pattern with characteristic peaks at diffraction angle 2 θ values of 6.5 ° ± 0.2 °, 16.3 ° ± 0.2 °, 19.8 ° ± 0.2 °,8.1 ° ± 0.2 °, 12.9 ° ± 0.2 °, 20.5 ° ± 0.2 °, 10.2 ° ± 0.2 °, 19.5 ° ± 0.2 ° at any of 1, or 2, or 3, or 4, or 5, or 6, or 7, or 8, using Cu-ka radiation.

Without limitation, the X-ray powder diffraction pattern of crystalline form CSIV is substantially as shown in figure 22.

Without limitation, crystalline form CSIV has a mass loss of about 2.5% when heated to 100 ℃, and the thermogravimetric analysis profile is substantially as shown in figure 24.

Without limitation, crystalline form CSIV has an exothermic peak at around 200 ℃ and an endothermic peak (onset temperature about 250 ℃) at around 253 ℃, which is a melting endothermic peak, as shown in fig. 25 by differential scanning calorimetry.

Without limitation, crystalline form CSIV is a hydrate.

Without limitation, crystalline form CSIV contains no more than 7% by mass of water.

Further, the crystal form CSIV contains not more than 4% by mass of water.

Further, the crystal form CSIV contains not more than 3% by mass of water.

According to an object of the present invention, the present invention also provides a preparation method of the crystalline form CSIV, the preparation method comprising:

mixing the solid of the compound I, methanesulfonic acid and an ether/water mixed solvent or an alcohol/water mixed solvent, stirring, separating the solid, and drying the solid at the temperature of more than 45 ℃.

Further, the molar charge ratio of the compound I solid to methanesulfonic acid is preferably 0.9:1 to 1.1:1, more preferably 1: 1; the ether solvent is preferably tetrahydrofuran; the volume ratio of the ether solvent to water is preferably 9: 1; the alcohol solvent is preferably isopropanol; the volume ratio of the alcohol solvent to the water is preferably 9: 1; the drying temperature is preferably 45 ℃ to 100 ℃, more preferably 50 ℃.

According to an object of the present invention, the present invention provides a crystalline form CSVI of the mesylate salt of compound I (hereinafter referred to as "crystalline form CSVI"). The X-ray powder diffraction pattern of crystalline form CSVI is substantially as shown in figure 28.

Without limitation, crystalline form CSVI is a 1, 4-dioxane solvate.

According to an object of the present invention, the present invention provides crystalline form K4 (hereinafter referred to as "crystalline form K4") of compound I mesylate. The X-ray powder diffraction pattern of form K4 is substantially as shown in figure 29.

On the one hand, the X-ray powder diffraction pattern of the crystal form K4 has characteristic peaks at diffraction angle 2 theta values of 7.5 degrees +/-0.2 degrees, 15.2 degrees +/-0.2 degrees and 17.5 degrees +/-0.2 degrees by using Cu-Kalpha radiation.

Further, using Cu-Ka radiation, the X-ray powder diffraction pattern of the crystal form K4 has characteristic peaks at 1, 2 or 3 of diffraction angle 2 theta values of 21.6 degrees +/-0.2 degrees, 22.8 degrees +/-0.2 degrees, 19.3 degrees +/-0.2 degrees; preferably, the X-ray powder diffraction pattern of the crystal form K4 has a characteristic peak at 3 points in diffraction angles 2 theta of 21.6 +/-0.2 degrees, 22.8 +/-0.2 degrees and 19.3 +/-0.2 degrees.

Further, using Cu-K α radiation, the X-ray powder diffraction pattern of form K4 has characteristic peaks at 1, or 2, or 3 of diffraction angle 2 θ values of 10.8 ° ± 0.2 °, 18.3 ° ± 0.2 °, 25.5 ° ± 0.2 °; preferably, the X-ray powder diffraction pattern of the crystal form K4 has a characteristic peak at 3 points with diffraction angles 2 theta of 10.8 +/-0.2 degrees, 18.3 +/-0.2 degrees and 25.5 +/-0.2 degrees.

On the other hand, the X-ray powder diffraction pattern of the crystal form K4 has characteristic peaks at any 1, or 2, or 3, or 4, or 5, or 6, or 7, or 8, or 9 of diffraction angle 2 theta values of 7.5 DEG + -0.2 °, 15.2 DEG + -0.2 °, 17.5 DEG + -0.2 °, 21.6 DEG + -0.2 °, 22.8 DEG + -0.2 °, 19.3 DEG + -0.2 °, 10.8 DEG + -0.2 °, 18.3 DEG + -0.2 °, 25.5 DEG + -0.2 ° using Cu-Ka radiation.

According to an object of the present invention, the present invention provides the use of the crystalline form CSII, the crystalline form CSIII, the crystalline form CSIV or any mixture of the above crystalline forms for the preparation of further crystalline forms of compound I methanesulfonic acid.

According to the purpose of the present invention, the present invention provides a pharmaceutical composition comprising an effective therapeutic amount of compound I mesylate in crystal form CSII, CSIII, CSIV or any mixture thereof, and pharmaceutically acceptable excipients.

According to the purpose of the invention, the invention provides application of a crystal form CSII, a crystal form CSIII, a crystal form CSIV or any mixture of the above crystal forms of the compound I mesylate in preparing ROCK2 inhibitor medicines.

According to an object of the present invention, there is provided a use of the crystalline form CSII, the crystalline form CSIII, the crystalline form CSIV or any mixture of the above crystalline forms of the mesylate salt of compound I for the preparation of a medicament for the treatment of chronic graft-versus-host disease, systemic sclerosis and idiopathic pulmonary fibrosis.

Advantageous effects

The crystal form CSII provided by the invention has the following advantages:

(1) compared with the prior art, the crystal form CSII provided by the invention has higher solubility. In FeSSIF, the solubility of crystalline form CSII is more than 2 times that of prior art P1 and 1.3 times that of prior art P2.

Compound I is a poorly water soluble drug and belongs to BCS IV. The crystal form CSII provided by the invention has higher solubility, is beneficial to improving the absorption of the medicine in a human body and improves the bioavailability; in addition, the higher solubility can ensure the curative effect of the medicine and reduce the dosage of the medicine, thereby reducing the side effect of the medicine and improving the safety of the medicine.

(2) Compared with the prior art, the crystal form CSII preparation provided by the invention has better in-vitro dissolution rate. The dissolution of the crystal form CSII preparation and the prior art preparation in ABS with pH 4.5 and PBS medium with pH 6.8 reaches balance within 120 minutes, and the average cumulative dissolution amount of the crystal form CSII preparation is 2 times of that of the prior art preparation.

Dissolution is an important prerequisite for absorption of the drug. Different crystal forms of the medicine can cause different dissolution in vivo, and finally cause difference of clinical efficacy. In vitro dissolution can predict the in vivo performance of a drug according to BCS guidelines. The crystal form CSII medicine provided by the invention has better in-vitro dissolution rate, indicates that the crystal form CSII medicine has higher in-vivo absorption degree and better exposure characteristic, thereby improving the bioavailability and the curative effect of the medicine.

(3) Compared with the prior art, the crystal form CSII provided by the invention has lower hygroscopicity. Test results show that the crystal form of the crystal form CSII of the invention remains unchanged before and after the hygroscopicity test, and the hygroscopicity increase (40-80% RH) is about 1/5 of P1 in the prior art. The hygroscopicity of the crystal form CSII is obviously superior to that of the prior art.

On one hand, the high hygroscopicity easily causes chemical degradation and solid form transformation of the bulk drugs, thereby directly influencing the physicochemical stability of the bulk drugs. In addition, high hygroscopicity can reduce the fluidity of the bulk drug, thereby affecting the processing technology of the bulk drug.

On the other hand, a drug with high hygroscopicity is required to maintain low humidity during production and storage, and thus, higher demands are made on production, which requires high cost. More importantly, the high hygroscopicity easily causes the content change of the effective components in the medicine, and influences the quality of the medicine.

The crystal form CSII provided by the invention has low hygroscopicity, does not have harsh requirements on the environment, reduces the production, storage and quality control costs of materials, and has strong economic value.

(4) The crystal form CSII bulk drug and the preparation provided by the invention have good stability. The crystal form CSII bulk drug is placed at 25 ℃/60% RH, the crystal form is not changed for at least 6 months, the chemical purity is more than 99.5%, and the purity is basically kept unchanged in the storage process. After the crystal form CSII and the auxiliary materials are mixed to prepare the medicinal preparation, the medicinal preparation is placed under the condition of 25 ℃/60 percent RH, the crystal form is not changed for at least one month, the chemical purity is more than 99.4 percent, and the purity is basically kept unchanged in the storage process. The crystal form CSII bulk drug and the preparation have better stability under long-term conditions, and are beneficial to the storage of the drug.

Meanwhile, the crystal form of the CSII bulk drug is not changed after being placed for at least 6 months under the condition of 40 ℃/75% RH, the chemical purity is more than 99.5%, and the purity is basically kept unchanged in the storage process. After the crystal form CSII and the auxiliary materials are mixed to prepare the medicinal preparation, the medicinal preparation is placed under the condition of 40 ℃/75% RH, the crystal form is not changed for at least one month, the chemical purity is more than 99.4%, and the purity is basically kept unchanged in the storage process. The crystal form CSII bulk drug and the preparation have good stability under the acceleration condition.

The good physical and chemical stability of the crystal form of the bulk drug can ensure that the drug does not generate crystal transformation and basically has no impurity generation in the production and storage processes. The crystal form CSII has good physical and chemical stability, ensures the quality of the raw material medicines and the preparation to be consistent and controllable, and reduces the medicine quality change, the bioavailability change and the toxic and side effects caused by crystal form change or impurity generation.

Meanwhile, the crystal form CSII has better grinding stability. In the prior art, the crystal form of the P1 ground bulk drug is basically converted into amorphous, but the crystal form of the CSII bulk drug of the invention remains unchanged after grinding, and the CSII bulk drug has good physical stability. The raw material medicines are usually required to be ground and crushed in the preparation processing process, and the good physical stability can reduce the risks of reduction of the crystallinity and crystal transformation of the raw material medicines in the preparation processing process.

(5) Compared with the prior art, the crystal form CSII provided by the invention has almost no solvent residue, while the prior art P2 contains about 0.34 molar equivalent of ethyl acetate (about 62000 ppm).

According to the ICH guidelines, ethyl acetate belongs to class 3 solvents with an upper permissible limit of 5000 ppm. Therefore, the solvent content of the P2 in the prior art is far beyond the upper limit, and is not suitable for medicinal use.

The crystal form CSIII provided by the invention has the following advantages:

(1) compared with the prior art, the crystal form CSIII provided by the invention has higher solubility. In FeSSIF, the solubility of crystalline form CSIII is more than 2 times that of prior art P1 and 1.7 times that of prior art P2.

Compound I is a poorly water soluble drug and belongs to BCS IV. The crystal form CSIII provided by the invention has higher solubility, is beneficial to improving the absorption of the medicine in a human body, and improves the bioavailability; in addition, the higher solubility can ensure the curative effect of the medicine and reduce the dosage of the medicine, thereby reducing the side effect of the medicine and improving the safety of the medicine.

(2) Compared with the prior art, the crystal form CSIII preparation provided by the invention has better in-vitro dissolution rate. The dissolution of the crystal form CSIII preparation and the prior art preparation in an ABS medium with the pH value of 4.5 reaches the balance within 120 minutes, and the average accumulated dissolution amount of the crystal form CSIII preparation is about 5 times of that of the prior art preparation. The dissolution of the crystal form CSIII preparation and the prior art preparation in PBS medium with pH 6.8 reaches the balance within 120 minutes, and the average accumulated dissolution amount of the crystal form CSIII preparation is about 3 times of that of the prior art preparation.

Dissolution is an important prerequisite for absorption of the drug. Different crystal forms of the medicine can cause different dissolution in vivo, and finally cause difference of clinical efficacy. In vitro dissolution may predict the in vivo performance of a drug according to BCS guidelines. The crystal form CSIII preparation provided by the invention has better in-vitro dissolution rate, indicates that the in-vivo absorption degree is higher, and the exposure characteristic is better, so that the bioavailability is improved, and the curative effect of the medicine is improved.

(3) Compared with the prior art, the crystal form CSIII provided by the invention has lower hygroscopicity. Test results show that the crystal form of the crystal form CSIII of the invention remains unchanged before and after the hygroscopicity test, and the hygroscopicity increase (0-80% RH) is about 1/3 of P1 in the prior art. The hygroscopicity of the crystal form CSIII is obviously superior to that of the prior art.

The crystal form CSIII provided by the invention has low hygroscopicity, does not have strict requirements on environment, reduces the production, storage and quality control costs of materials, and has very strong economic value.

(4) The crystal form CSIII bulk drug and the preparation provided by the invention have good stability. The crystal form CSIII bulk drug is placed at 25 ℃/60% RH, the crystal form is not changed for at least 6 months, the chemical purity is more than 99.7%, and the purity is basically kept unchanged in the storage process. After the crystal form CSIII is mixed with auxiliary materials to be prepared into a medicinal preparation, the medicinal preparation is placed under the condition of 25 ℃/60% RH, the crystal form is unchanged for at least one month, the chemical purity is more than 99.5%, and the purity is basically kept unchanged in the storage process. The crystal form CSIII bulk drug and the preparation have better stability under long-term conditions, and are beneficial to the storage of the drug.

Meanwhile, the crystal form of the CSIII bulk drug is not changed after being placed for at least 6 months under the condition of 40 ℃/75% RH, the crystal form is not changed for at least one month under the condition of 60 ℃/75% RH, the chemical purity is more than 99.7%, and the purity is basically kept unchanged in the storage process. After the crystal form CSIII is mixed with auxiliary materials to be prepared into a medicinal preparation, the medicinal preparation is placed under the condition of 40 ℃/75% RH, the crystal form is unchanged for at least one month, the chemical purity is more than 99.5%, and the purity is basically kept unchanged in the storage process. The crystal form CSIII bulk drug and the preparation have good stability under the acceleration condition and the harsher condition.

High temperature and high humidity conditions caused by seasonal differences, climate differences in different regions, environmental factors and the like can influence the storage, transportation and production of the raw material medicines. Therefore, the stability of bulk drugs and formulations under accelerated and more severe conditions is of great importance for drugs. The crystal form CSIII bulk drug and the preparation have better stability under harsh conditions, and are beneficial to avoiding the influence on the quality of the drug caused by crystal transformation or purity reduction in the storage process of the drug.

The good physical and chemical stability of the crystal form of the bulk drug can ensure that the drug is not subjected to crystal transformation and basically no impurities are generated in the production and storage processes. The crystal form CSIII has good physical and chemical stability, ensures the quality of the raw material medicines and the preparation to be consistent and controllable, and reduces the medicine quality change, the bioavailability change and the toxic and side effects caused by crystal form change or impurity generation.

Meanwhile, the crystal form CSIII has better grinding stability. The crystal form of the CSIII crystal form bulk drug of the invention remains unchanged after being ground, and the P1 crystal form bulk drug of the invention has good physical stability. The raw material medicines are usually ground and crushed in the preparation processing process, and the good physical stability can reduce the risks of reduction of the crystallinity and crystal transformation of the raw material medicines in the preparation processing process.

(5) Compared with the prior art, the crystal form CSIII provided by the invention has almost no solvent residue, while the prior art P2 contains about 0.34 molar equivalent of ethyl acetate (about 62000 ppm).

The crystal form CSIV provided by the invention has the following advantages:

(1) compared with the prior art, the crystal form CSIV provided by the invention has higher solubility. In FeSSIF, the solubility of crystalline CSIV is more than 2 times that of prior art P1 and 1.4 times that of prior art P2.

Compound I is a poorly water soluble drug and belongs to BCS IV. The crystal form CSIV provided by the invention has higher solubility, is beneficial to improving the absorption of the medicine in a human body and improves the bioavailability; in addition, the higher solubility can ensure the curative effect of the medicine and reduce the dosage of the medicine, thereby reducing the side effect of the medicine and improving the safety of the medicine.

(2) The crystal form CSIV bulk drug provided by the invention has good stability. The crystal form CSIV bulk drug is placed at 25 ℃/60% RH, the crystal form is not changed for at least 3 months, the chemical purity is more than 99.5%, and the purity is basically kept unchanged in the storage process. The crystal form CSIV bulk drug has better stability under long-term conditions and is beneficial to the storage of the drug.

Meanwhile, the crystal form of the CSIV bulk drug is not changed after being placed for at least 3 months under the condition of 40 ℃/75% RH, the crystal form is not changed for at least one month under the condition of 60 ℃/75% RH, the chemical purity is over 99.5 percent, and the purity is basically kept unchanged in the storage process. The crystal form CSIV bulk drug has good stability under the acceleration condition and the harsher condition.

High temperature and high humidity conditions caused by seasonal differences, climate differences in different regions, environmental factors and the like can influence the storage, transportation and production of the raw material medicines. Therefore, the stability of bulk drugs and formulations under accelerated and more severe conditions is of great importance for drugs. The crystal form CSIV bulk drug and the preparation have better stability under harsh conditions, and are beneficial to avoiding the influence on the quality of the drug caused by crystal transformation or purity reduction in the storage process of the drug.

The good physical and chemical stability of the crystal form of the bulk drug can ensure that the drug does not generate crystal transformation and basically has no impurity generation in the production and storage processes. The crystal form CSIV has good physical and chemical stability, ensures the quality of the raw material medicines and the preparation to be consistent and controllable, and reduces the medicine quality change, the bioavailability change and the toxic and side effects caused by crystal form change or impurity generation.

Meanwhile, the crystal form CSIV has better grinding stability. In the prior art, the crystal form of the P1 ground bulk drug is basically converted into amorphous, but the crystal form of the CSIV bulk drug of the invention remains unchanged after grinding, and the CSIV bulk drug has good physical stability. The raw material medicines are usually ground and crushed in the preparation processing process, and the good physical stability can reduce the risks of reduction of the crystallinity and crystal transformation of the raw material medicines in the preparation processing process.

(3) In contrast to the prior art, the present invention provides crystalline form CSIV with almost no solvent residue, whereas prior art P2 contains about 0.34 molar equivalents of ethyl acetate (about 62000 ppm).

Drawings

FIG. 1 is an XRPD pattern of prior art P1

FIG. 2 is an XRPD pattern of prior art P2

FIG. 3 is an XRPD pattern for crystalline form CSII

FIG. 4 is a TGA profile of crystalline form CSII

FIG. 5 is a DSC of crystalline form CSII

FIG. 6 is a comparison of XRPD patterns before and after grinding for prior art P1 (top: after grinding; bottom: before grinding)

FIG. 7 is a comparison of XRPD before and after milling for crystalline form CSII (top: after milling; bottom: before milling)

FIG. 8 is a comparison of XRPD before and after DVS testing for crystalline form CSII (top: before testing; bottom: after testing)

FIG. 9 is an XRPD comparison graph of the crystal form CSII before and after placement under different conditions (from bottom to top: before placement, 25 ℃/60% RH open package is placed for 6 months, 25 ℃/60% RH sealed package is placed for 6 months, 40 ℃/75% RH open package is placed for 6 months, 40 ℃/75% RH sealed package is placed for 6 months)

FIG. 10 shows XRPD patterns of the crystal form CSII and its preparation (from bottom to top: blank mixed powder, crystal form CSII preparation, crystal form CSII)

FIG. 11 is a comparison XRPD of a crystalline form of CSII formulation before and after placement under different conditions (from bottom to top: before placement, for one month at 25 ℃/60% RH and for one month at 40 ℃/75% RH)

FIG. 12 is an XRPD pattern for crystalline form CSIII

FIG. 13 is a TGA profile of crystalline form CSIII

FIG. 14 is a DSC of crystalline form CSIII

FIG. 15 is a comparison of XRPD before and after milling for crystalline form CSIII (top: after milling; bottom: before milling)

FIG. 16 is a DVS diagram of prior art P1

FIG. 17 is a DVS diagram of crystalline form CSIII

FIG. 18 is a comparison of XRPD before and after DVS testing for crystalline form CSIII (top: before testing; bottom: after testing)

FIG. 19 is an XRPD comparison graph of the crystal form CSIII before and after being placed under different conditions (from bottom to top: before being placed, 25 ℃/60% RH open package is placed for 6 months, 25 ℃/60% RH sealed package is placed for 6 months, 40 ℃/75% RH sealed package is placed for 6 months, 60 ℃/75% RH sealed package is placed for one month)

FIG. 20 is an XRPD pattern for crystalline form CSIII and its formulation (from bottom to top: blank powder, crystalline form CSIII formulation, crystalline form CSIII)

FIG. 21 is a comparison XRPD of a crystalline form of CSIII formulation before and after placement under different conditions (from bottom to top: before placement, for one month at 25 ℃/60% RH and for one month at 40 ℃/75% RH)

FIG. 22 is an XRPD pattern for crystalline form CSIV

FIG. 23 is an XRPD pattern for crystalline form CSIV

FIG. 24 is a TGA profile of crystalline form CSIV

FIG. 25 is a DSC of crystalline form CSIV

FIG. 26 is a comparison of XRPD before and after milling of crystalline form CSIV (top: after milling; bottom: before milling)

FIG. 27 is an XRPD comparison chart of the crystal form CSIV before and after being placed under different conditions (from bottom to top: before being placed, 25 ℃/60% RH sealed package is placed for 3 months, 40 ℃/75% RH sealed package is placed for 3 months, and 60 ℃/75% RH sealed package is placed for one month)

FIG. 28 is an XRPD pattern for crystalline form CSVI

FIG. 29 is an XRPD pattern for form K4

Detailed Description

The invention is illustrated in detail by the following examples describing in detail the methods of making and using the crystalline forms of the invention. It will be apparent to those skilled in the art that many modifications, both to materials and methods, may be practiced without departing from the scope of the invention.

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

XRPD: powder X-ray diffraction

DSC: differential scanning calorimetry

TGA: thermogravimetric analysis

DVS: dynamic moisture adsorption

¹ H NMR: liquid nuclear magnetic hydrogen spectrum

And (4) UPLC: ultra-high performance liquid chromatography

HPLC: high performance liquid chromatography

RH: relative humidity

BCS: biopharmaceutical classification system

ABS: acetate buffer solution

PBS: phosphate buffer

The instrument and method for data acquisition:

the X-ray powder diffractogram described in the examples of the present invention was collected on a Bruker X-ray powder diffractometer. The method parameters of X-ray powder diffraction are as follows:

an X-ray light source: cu, K alpha

1.54060；

1.54439

The K alpha 2/K alpha 1 intensity ratio: 0.50

Thermogravimetric analysis (TGA) profiles described herein were collected on TA Q500. The parameters of the thermogravimetric analysis method of the invention are as follows:

scanning rate: 10 ℃/min

Protective gas: n is a radical of ₂

Differential Scanning Calorimetry (DSC) profile described herein was taken on a TA Q2000 panel. The parameters of the differential scanning calorimetry analysis method are as follows:

scanning rate: 10 ℃/min

Protective gas: n is a radical of ₂

The dynamic moisture sorption (DVS) profile of the present invention was collected on an Intrinsic dynamic moisture sorption instrument manufactured by SMS corporation (Surface Measurement Systems Ltd.). The instrument control software is DVS-Intrasic control software. The method parameters of the dynamic moisture adsorption instrument are as follows:

temperature: 25 deg.C

Carrier gas, flow rate: n is a radical of ₂ 200 ml/min

Relative humidity range: 0% RH-95% RH

Nuclear magnetic resonance hydrogen spectrum data of the present invention ( ¹ H NMR) was taken from Bruker Avance II DMX 400MHz NMR spectrometer. Weighing 1-5mg sample, dissolving with 0.5mL deuterated dimethyl sulfoxide or deuterated methanol to prepare 2-10mg/mL solution.

The solubility test method of the present invention is shown in table 1.

TABLE 1

The method for detecting the substance according to the present invention is shown in Table 2.

TABLE 2

The dissolution rate detection method of the present invention is shown in table 3.

TABLE 3

In the present invention, the "stirring" is performed by a conventional method in the art, such as magnetic stirring or mechanical stirring, and the stirring speed is 50-1800 rpm, wherein the magnetic stirring speed is preferably 300-900 rpm, and the mechanical stirring speed is preferably 100-300 rpm.

The "separation" is accomplished by methods conventional in the art, such as centrifugation or filtration. The operation of "centrifugation" was: and placing the sample to be separated into a centrifuge tube, centrifuging at the speed of 10000 rpm until all solids are settled to the bottom of the centrifuge tube, and separating the solids.

The "drying" is carried out by methods conventional in the art, such as vacuum drying, forced air drying or natural air drying. The drying temperature may be room temperature or higher, preferably room temperature to about 60 ℃, alternatively to 50 ℃, alternatively to 40 ℃. The drying time may be 2-48 hours, or overnight. Drying is carried out in a fume hood, a forced air oven or a vacuum oven.

The 'sealed package' is completed by adopting a conventional method in the field, for example, a sample is placed in a glass small bottle, the mouth of the bottle is tightly screwed by a bottle cap, and finally, the bottle is sealed in an aluminum foil bag; or placing the sample in a glass small bottle, tightly screwing the bottle mouth with a bottle cap, and then sealing the bottle mouth and 1g of silica gel desiccant in an aluminum foil bag; or the sample is placed in a glass small bottle, the mouth of the bottle is covered with a layer of aluminum foil paper, a hole is formed in the aluminum foil paper, and then the aluminum foil paper and 1g of silica gel desiccant are sealed in an aluminum foil bag.

The "open pack" is accomplished by methods conventional in the art, such as placing the sample in a glass vial, capping the vial with a foil and forming an opening in the foil.

The "room temperature" is not a specific temperature value, and means a temperature range of 10 to 30 ℃.

The "characteristic peak" refers to a representative diffraction peak used for screening crystals, and the peak position can usually have an error of ± 0.2 ° when tested using Cu — K α radiation.

In the present invention, "crystal" or "crystal form" can be characterized by X-ray powder diffraction. Those skilled in the art will appreciate that the X-ray powder diffraction pattern will vary depending on the conditions of the instrument, sample preparation and sample purity. The relative intensities of diffraction peaks in an X-ray powder diffraction pattern may also vary with experimental conditions, so that the intensities of diffraction peaks cannot be the only or decisive factor for determining the crystal form. In fact, the relative intensities of the diffraction peaks in the X-ray powder diffraction pattern are related to the preferred orientation of the crystals, and the intensities of the diffraction peaks shown in the present invention are illustrative and not used for absolute comparison. Thus, it will be understood by those skilled in the art that the X-ray powder diffraction patterns of the crystalline forms of the invention do not have to be identical to the X-ray powder diffraction patterns of the examples referred to herein, and any crystalline form having an X-ray powder diffraction pattern identical or similar to the characteristic peaks in these patterns is within the scope of the invention. One skilled in the art can compare the X-ray powder diffraction pattern listed in the present invention with an X-ray powder diffraction pattern of an unknown crystalline form to confirm whether the two sets of patterns reflect the same or different crystalline forms.

In some embodiments, the crystalline form CSII, the crystalline form CSIII, the crystalline form CSIV, the crystalline form CSVI, the crystalline form K4 of the present invention are pure, substantially free of any other crystalline forms in admixture. As used herein, "substantially free" when used in reference to a novel form means that the form contains less than 20% by weight of the other form, particularly less than 10% by weight of the other form, more particularly less than 5% by weight of the other form, and even more particularly less than 1% by weight of the other form.

The term "about" when used in reference to a measurable quantity, such as mass, time, temperature, etc., means a range that can float around the specified quantity, which range can be 10%, 5%, 1%, 0.5%, or 0.1%.

The following examples were conducted at room temperature unless otherwise specified.

According to the present invention, the compound I and/or a salt thereof as a starting material includes, but is not limited to, a solid form (crystalline or amorphous), an oil form, a liquid form and a solution. Preferably, compound I and/or its salt as starting material is in solid form.

The compounds I used in the examples below can be prepared according to the prior art, for example according to the method described in WO2006105081A 2.

Example 1 prior art repetition and characterization

Process for the preparation of compound I according to WO2006105081A2An XRPD pattern of prior art P1 was obtained, and of prior art P1 is shown in fig. 1. According to the preparation method of compound I described in CN106916145B, prior art P2 was obtained, and the XRPD pattern of prior art P2 is shown in fig. 2. Of the solid ¹ The H NMR data are: ¹ h NMR (400MHz, DMSO-d6) δ 13.09(s,1H),9.96(s,1H),8.59(d, J ═ 8.3Hz,1H),8.31(d, J ═ 1.8Hz,1H),8.15(s,1H),8.09-7.98(m,2H),7.98-7.79(m,4H),7.69-7.56(m,2H),7.41(t, J ═ 7.9Hz,1H),7.08(ddd, J ═ 8.2,2.7,1.0Hz,1H),4.51(s,2H),4.03(m, J ═ 7.1Hz,1.62H),1.99(s,1.02H),1.17(t,1.07H),1.11(d, 6.03 (m, J ═ 7.1Hz, 1H), and ethyl acetate where the nuclear magnetic resonance signals at the ethyl acetate and acetic acid overlap at ppm. The resulting prior art P2 was calculated to contain about 0.34 molar equivalents of ethyl acetate (about 62000 ppm). According to the ICH guidelines, ethyl acetate belongs to class 3 solvents, the allowable upper limit of which is 5000ppm, and the content of ethyl acetate in prior art P2 is much higher than the upper limit of the ICH guidelines, and is not suitable for pharmaceutical use.

Example 2 preparation of crystalline form CSII

298.2mg of a solid of Compound I was weighed in a vial, 6mL of a mixed solvent of tetrahydrofuran/water (9:1, v/v) and 44. mu.L of a methanesulfonic acid liquid were sequentially added, stirred at room temperature for about 2 days, the solid was separated, and the resulting solid was dried under vacuum at 25 ℃ for about 2 hours to obtain a crystalline solid.

Through detection, the obtained crystalline solid is the crystal form CSII, the X-ray powder diffraction pattern is shown in figure 3, and the X-ray powder diffraction data is shown in table 4.

TGA of crystalline form CSII is shown in figure 4 with about 5.9% mass loss when heated to 100 ℃.

DSC shows that an endothermic peak exists at about 100 ℃ as shown in FIG. 5; an exothermic peak exists around 206 ℃; an endothermic peak (onset temperature about 253 ℃) was present near 255 ℃.

Nuclear magnetic data for crystalline form CSII were: ¹ H NMR(400MHz,DMSO-d6)δ13.26(s,1H),11.37(s,1H),8.74(d,J＝8.3Hz,1H),8.20(d,J＝12.8Hz,2H),8.08(dt,J＝15.8,8.2Hz,2H),7.93(d,J＝8.0Hz,1H),7.90–7.80(m,3H),7.80–7.66(m,2H),7.55(t,J＝8.0Hz,1H),7.34–7.18(m,1H),4.54(s,2H),3.95(dq, J ═ 13.6,6.8Hz,1H),2.33(s,3H),1.09(d, J ═ 6.6Hz,6H). According to nuclear magnetic data, the molar ratio of the compound I to the methanesulfonic acid in the crystal form CSII is 1: 1.

TABLE 4

Example 3 preparation of crystalline form CSII

29.5mg of Compound I solid was weighed into a vial, and 0.6mL of a mixed solvent of isopropanol/water (9:1, v/v) and 4.4. mu.L of methanesulfonic acid liquid were sequentially added, stirred at room temperature for about 3 days, and the solid was isolated. Through detection, the obtained solid is the crystal form CSII, and X-ray powder diffraction data of the solid is shown in Table 5.

TABLE 5

Example 4 solubility of crystalline form CSII and Prior Art

About 10mg of each of the crystal form CSII of the present invention and the solid of the prior art was dispersed in 0.8mL of FeSSIF to prepare a suspension, equilibrated at 37 ℃ for 15 minutes, filtered to obtain a clear solution, and the content (μ g/mL) of the mesylate of compound I in the solution was measured by UPLC, with the results shown in Table 6.

TABLE 6

Crystal form	Concentration in FeSSIF (μ g/mL)
		Crystal form CSII	75.07
Prior art P1	35.93
		Prior art P2	57.26

The result shows that compared with the prior art, the crystal form CSII has higher solubility in FeSSIF.

Example 5 grinding stability of crystalline form CSII and Prior Art

The crystalline form CSII and the prior art solid were separately placed in a mortar, hand milled for 5 minutes, and the samples were tested for XRPD before and after milling, with the results shown in table 7. The results show that the crystalline form CSII has better grinding stability compared to the prior art.

TABLE 7

Before grinding	After grinding	XRPD comparison before and after grinding
			Prior art P1	Conversion to substantially amorphous	FIG. 6
Crystal form CSII	The crystal form remains unchanged	FIG. 7

EXAMPLE 6 crystalline form CSII and hygroscopicity of the prior art

The prior art solid and the crystalline form CSII of the present invention were weighed to about 10mg each and tested for hygroscopicity using a dynamic moisture sorption (DVS) instrument. The mass change at each humidity was recorded at the corresponding humidity cycle. The results of the experiment are shown in Table 8.

TABLE 8

Crystal form	Humidity cycling	Mass variation (40-80% RH)
			Prior art P1	0-95-0％RH	1.11％
Crystal form CSII	40-95-0-95％RH	0.23％

Note: the crystalline form CSII was a hydrate and the DVS test started with ambient humidity (40% RH).

Experimental results show that the hygroscopicity of the crystal form CSII is increased by 0.23% under the condition of 40-80% RH, the hygroscopicity of the solid in the prior art is increased by 1.11% under the condition of 40-80% RH, the hygroscopicity of the crystal form CSII is increased by only 1/5 in the prior art, and the hygroscopicity of the crystal form CSII is obviously superior to that of the prior art.

Example 7 humidity stability of crystalline form CSII

Form CSII was tested for XRPD before and after DVS testing and the results are shown in figure 8. The experimental results show that the crystal form of the CSII is kept unchanged after 40-95-0-95% RH cycles. The crystal form CSII has good stability under high-humidity and low-humidity conditions.

EXAMPLE 8 physicochemical stability of crystalline form CSII

Taking a proper amount of the crystal form CSII, respectively placing for a certain time under the conditions of 25 ℃/60% RH and 40 ℃/75% RH, and determining the purity and the crystal form by using UPLC and XRPD. The results are shown in Table 9, and the XRPD pattern is shown in FIG. 9.

TABLE 9

The experimental result shows that the crystal form CSII can be stable for at least 6 months under the conditions of 25 ℃/60% RH and 40 ℃/75% RH, and the crystal form CSII has good stability under long-term and accelerated conditions.

EXAMPLE 9 preparation of Compound I preparation

1. Formulation formula

Watch 10

2. Preparation process

TABLE 11

Example 10 formulation stability of crystalline form CSII

Form CSII was not changed after being prepared into tablets according to example 9. The XRPD pattern of the crystalline form CSII and its formulation is shown in figure 10.

EXAMPLE 11 dissolution of crystalline form CSII formulation and Prior Art formulation

After the crystalline form CSII and prior art P1 were prepared into tablets according to example 9, they were tested for in vitro dissolution. The determination of dissolution is carried out according to 0931 dissolution and release determination methods in the Chinese pharmacopoeia 2020 edition, and the test conditions are shown in Table 12.

TABLE 12

The experimental results show that in ABS with pH 4.5 and PBS with pH 6.8, the dissolution reaches the equilibrium within 120 minutes, and the average accumulated dissolution amount of the crystal form CSII preparation is about 2 times of that of the preparation in the prior art. Therefore, the crystal form CSII has obvious in-vivo bioavailability advantages compared with the prior art.

EXAMPLE 12 stability of crystalline form CSII formulation

The crystalline form CSII formulation prepared according to example 9 was placed at 25 ℃/60% RH and 40 ℃/75% RH and tested for crystalline form and purity after 1 month. The test results are shown in fig. 11 and table 13.

Watch 13

The results show that the crystal form CSII preparation is kept unchanged and the purity is basically unchanged after being placed for one month under the conditions of 25 ℃/60% RH and 40 ℃/75% RH. The preparation of the crystal form CSII has good stability under long-term and accelerated conditions.

Example 13 preparation of crystalline form CSIII

298.2mg of a solid of compound I was weighed in a vial, 6mL of a mixed solvent of tetrahydrofuran/water (9:1, v/v) and 44. mu.L of a methanesulfonic acid liquid were sequentially added, stirred at room temperature for about 2 days, the solid was separated, and after drying the solid at 25 ℃ in vacuo for about 2 hours, the solid was heated to 215 ℃ under nitrogen atmosphere and kept at the constant temperature for 1 minute, a crystalline solid was obtained.

Through detection, the obtained crystalline solid is the crystal form CSIII, the X-ray powder diffraction pattern is shown in figure 12, and the X-ray powder diffraction data is shown in table 14.

The TGA of crystalline form CSIII is shown in figure 13 and has a mass loss of about 0.1% when heated to 200 ℃.

DSC shows an endothermic peak (onset temperature of about 252 ℃ C.) at about 253 ℃ C as a melting endothermic peak in FIG. 14.

The nuclear magnetic data for crystalline form CSIII are: ¹ h NMR (400MHz, DMSO) δ 13.25(s,1H),11.31(s,1H),8.73(d, J ═ 8.1Hz,1H),8.20(d, J ═ 10.2Hz,2H), 8.13-8.00 (m,2H),7.93(d, J ═ 8.0Hz,1H),7.85(d, J ═ 9.0Hz,3H),7.75(dt, J ═ 23.2,5.3Hz,2H),7.55(s,1H),7.26(s,1H),4.54(s,2H),3.95(d, J ═ 7.6Hz,1H),2.32(s,3H),1.09(d, J ═ 6.6Hz,6H). According to nuclear magnetic data, the molar ratio of the compound I to methanesulfonic acid in the crystal form CSIII is 1: 1.

TABLE 14

Example 14 solubility of crystalline form CSIII and Prior Art

About 10mg of each of the crystal form CSIII of the present invention and the solid of the prior art was dispersed in 0.8mL of FeSSIF to prepare a suspension, equilibrated at 37 ℃ for 15 minutes, filtered to obtain a clear solution, and the content (μ g/mL) of the compound I mesylate in the solution was tested by UPLC, with the results shown in Table 15.

Watch 15

Crystal form	Concentration in FeSSIF (μ g/mL)
		Crystal form CSIII	99.35
Prior art P1	35.93
		Prior art P2	57.26

The results show that the crystal form CSIII has higher solubility compared with the prior art.

EXAMPLE 15 grinding stability of crystalline form CSIII and Prior Art

The crystalline form CSIII and the prior art solid were separately placed in a mortar, hand milled for 5 minutes, and the samples were tested for XRPD before and after milling, with the results shown in table 16. The results show that the crystalline form CSIII has better milling stability compared to the prior art.

TABLE 16

Before grinding	After grinding	XRPD contrast before and after grinding
			Prior art P1	Conversion to substantially amorphous	FIG. 6
Crystal form CSIII	The crystal form remains unchanged	FIG. 15 shows a schematic view of a

EXAMPLE 16 crystalline form CSIII and hygroscopicity of the prior art

The prior art solid and the crystalline form CSIII of the present invention were weighed to about 10mg each and tested for hygroscopicity using a dynamic moisture sorption (DVS) instrument. The mass change at each humidity was recorded at the corresponding humidity cycle. The results of the experiment are shown in Table 17.

TABLE 17

Crystal form	Humidity cycling	Mass variation (0-80% RH)	DVS diagram
				Prior art P1	0-95-0％RH	1.32％	FIG. 16
Crystal form CSIII	0-95-0％RH	0.40％	FIG. 17

Experimental results show that the hygroscopicity increase of the crystal form CSIII under the condition of 0-80% RH is 0.40%, the hygroscopicity increase of the crystal form P1 under the condition of 0-80% RH is 1.32%, the hygroscopicity increase of the crystal form CSIII is only 1/3 of the hygroscopicity increase of the crystal form in the prior art, and the hygroscopicity of the crystal form CSIII is obviously superior to that of the crystal form in the prior art.

Example 17 moisture stability of crystalline form CSIII

Form CSIII was tested for XRPD before and after DVS testing and the results are shown in figure 18. The experimental results show that the crystal form of the crystal form CSIII remains unchanged after 0-95-0% RH cycles. The crystal form CSIII has good stability under high-humidity and low-humidity conditions.

EXAMPLE 18 physicochemical stability of crystalline form CSIII

Taking a proper amount of the crystal form CSIII prepared by the invention, respectively placing for a period of time under the conditions of 25 ℃/60% RH, 40 ℃/75% RH and 60 ℃/75% RH, and determining the purity and the crystal form by UPLC and XRPD. The results are shown in Table 18, and the XRPD pattern is shown in FIG. 19.

Watch 18

The experimental result shows that the crystal form CSIII can be stable for at least 6 months under the conditions of 25 ℃/60% RH and 40 ℃/75% RH, and the crystal form CSIII can keep good stability under long-term and accelerated conditions. The crystal form CSIII can be stable for at least 1 month under the condition of 60 ℃/75% RH, and the stability of the crystal form CSIII is also good under the severer condition.

EXAMPLE 19 formulation stability of crystalline form CSIII

Form CSIII was prepared as tablets according to example 9 without modification. The XRPD contrast pattern of the crystalline form CSIII and its formulation is shown in figure 20.

EXAMPLE 20 dissolution of crystalline form CSIII formulation and Prior Art formulation

Form CSIII and prior art P1 were each tested for in vitro solubility after being prepared into tablets according to example 9. The determination of dissolution is carried out according to 0931 dissolution and release determination methods in Chinese pharmacopoeia 2020 edition, and the test conditions are shown in Table 12.

The experimental results show that the dissolution reaches the equilibrium within 120 minutes, the average cumulative dissolution amount of the crystal form CSIII preparation in ABS with pH 4.5 is about 5 times that of the prior art preparation, and the average cumulative dissolution amount of the crystal form CSIII preparation in PBS with pH 6.8 is about 3 times that of the prior art preparation. Therefore, the crystal form CSIII has obvious in-vivo bioavailability advantages compared with the prior art.

EXAMPLE 21 stability of crystalline form CSIII formulation

The crystalline CSIII formulation prepared according to example 9 was placed at 25 ℃/60% RH and 40 ℃/75% RH and tested for crystalline form and purity after 1 month. The test results are shown in fig. 21 and table 19.

Watch 19

The results show that the crystal form of the CSIII preparation is kept unchanged and the purity is basically unchanged after the CSIII preparation is placed for one month under the conditions of 25 ℃/60% RH and 40 ℃/75% RH. The preparation of the crystal form CSIII has good stability under long-term and accelerated conditions.

EXAMPLE 22 preparation of crystalline form CSIV

15.2mg of a solid of Compound I was weighed in a vial, 0.3mL of a mixed solvent of tetrahydrofuran/water (9:1, v/v) and 2.2. mu.L of a methanesulfonic acid liquid were added, stirred at room temperature for about 3 days, the solid was separated, and the obtained solid was dried under vacuum at 50 ℃ for 3 hours to obtain a crystalline solid.

Through detection, the obtained crystalline solid is the crystal form CSIV, the X-ray powder diffraction pattern is shown in figure 22, and the X-ray powder diffraction data is shown in table 20.

Watch 20

EXAMPLE 23 preparation of crystalline form CSIV

29.5mg of a solid of Compound I was weighed in a vial, 0.6mL of a mixed solvent of isopropanol/water (9:1, v/v) and 4.4. mu.L of methanesulfonic acid liquid was added, stirred at room temperature for about 3 days, the solid was separated, and the obtained solid was dried under vacuum at 50 ℃ for 3 hours to obtain a crystalline solid.

Through detection, the obtained crystalline solid is the crystal form CSIV, and X-ray powder diffraction data of the crystalline solid are shown in figure 23 and table 21.

TGA is shown in figure 24, having about 2.5% mass loss when heated to 100 ℃.

¹ The H NMR data are: ¹ h NMR (400MHz, DMSO-d6) δ 13.26(s,1H),11.40(s,1H),8.74(d, J ═ 8.3Hz,1H), 8.27-8.15 (m,2H),8.08(dt, J ═ 16.5,8.1Hz,2H),7.93(d, J ═ 8.0Hz,1H),7.85(q, J ═ 7.9Hz,3H), 7.80-7.68 (m,2H),7.56(t, J ═ 8.0Hz,1H),7.27(dd, J ═ 8.4,2.5Hz,1H),4.54(s,2H), 4.01-3.89 (m,1H),2.33(s,3H),1.09(d, J ═ 6.6H), 6H), methanesulfonic acid molar ratio of the compound to 1H, CSIV, and 1 molar ratio of the methanesulfonic acid.

TABLE 21

EXAMPLE 24 DSC of form CSIV

A sample of crystalline form CSIV was taken and subjected to DSC measurement, as shown in FIG. 25, which shows an exothermic peak at around 200 ℃ and an endothermic peak at around 253 ℃ (initial temperature of about 250 ℃), which is a melting endothermic peak.

EXAMPLE 25 solubility of crystalline form CSIV and Prior Art

About 10mg of each of the crystal form CSIV of the invention and the solid in the prior art is taken and respectively dispersed in 0.8mL of FeSSIF to prepare suspension, the suspension is balanced for 15 minutes at 37 ℃, then the suspension is filtered to obtain clear solution, and the UPLC is used for testing the content (mu g/mL) of the sample in the solution, and the results are shown in Table 22.

TABLE 22

Crystal form	Concentration in FeSSIF (μ g/mL)
		Crystal form CSIV	81.21
Prior art P1	35.93
		Prior art P2	57.26

The result shows that compared with the prior art, the crystal form CSIV has higher solubility in FeSSIF.

EXAMPLE 26 grinding stability of crystalline form CSIV and Prior Art

The crystalline CSIV prior art solid was separately placed in a mortar and hand milled for 5 minutes and the samples were tested for XRPD before and after milling with the results shown in table 23. The results show that the crystalline form CSIV has better milling stability compared to the prior art.

TABLE 23

Before grinding	After grinding	XRPD comparison before and after grinding
			Prior art P1	Conversion to substantially amorphous	FIG. 6
Crystal form CSIV	The crystal form remains unchanged	FIG. 26

EXAMPLE 27 physicochemical stability of crystalline form CSIV

Taking a proper amount of the crystal form CSIV prepared by the invention, respectively placing for a period of time under the conditions of 25 ℃/60% RH, 40 ℃/75% RH and 60 ℃/75% RH, and determining the purity and the crystal form by UPLC and XRPD. The results are shown in Table 24, and the XRPD contrast is shown in FIG. 27.

TABLE 24

The experimental result shows that the crystal form CSIV can be stable for at least 3 months under the conditions of 25 ℃/60% RH and 40 ℃/75% RH, and the crystal form CSIV can keep good stability under the long-term and accelerated conditions. The crystal form CSIV can be stable for at least 1 month under the condition of 60 ℃/75% RH, and the stability of the crystal form CSIV is very good under the severer condition.

EXAMPLE 28 preparation of crystalline form CSVI

202.2mg of Compound I solid was weighed into a vial, 4mL of 1, 4-dioxane and 30 μ L of methanesulfonic acid liquid were added, stirred at room temperature for about 9 days, the solid was isolated and dried under vacuum at 25 ℃ for 2 hours to give a crystalline solid.

The obtained crystalline solid is detected to be the crystal form CSVI, and the X-ray powder diffraction data of the crystalline solid is shown in figure 28 and table 25.

The nuclear magnetic data are as follows: ¹ H NMR(400MHz,Methanol-d4)δ8.64(d,J＝8.4Hz,1H),8.22–8.10(m,3H),8.09–8.02(m,1H),7.94–7.71(m,5H),7.55(t, J ═ 8.0Hz,1H), 7.39-7.31 (m,1H),4.54(s,2H), 4.15-4.01 (m,1H),3.66(s,6H),2.70(s,3H),1.15(d, J ═ 6.6Hz,6H), with a 1.4-dioxane nmr signal at 3.66 ppm. Form K3 was calculated to contain about 0.8 molar equivalents of 1, 4-dioxane (about 114000 ppm).

TABLE 25

Example 29 preparation of form K4

The solid obtained in example 28 was heated to 90 ℃ under nitrogen and kept at the same temperature for 2 minutes, and cooled to room temperature to obtain a crystalline solid.

XRPD testing of the resulting solid (test environment humidity below 45% RH) indicated that the resulting crystalline solid was crystalline form K4 according to the present invention, with X-ray powder diffraction data as shown in figure 29, table 26.

Watch 26

The crystal form K4 is a crystal form with very poor physical stability, can generate crystal transformation after being placed for only one week under the conditions of 40 ℃/75% RH and 60 ℃/75% RH, and is not suitable for medicinal use.

The above embodiments are merely illustrative of the technical ideas and features of the present invention, and the purpose thereof is to enable those skilled in the art to understand the contents of the present invention and implement the present invention, and not to limit the protection scope of the present invention. All equivalent changes and modifications made according to the spirit of the present invention should be covered within the protection scope of the present invention.

Claims

1. A crystal form of the mesylate of the compound I, which is characterized in that an X-ray powder diffraction pattern has characteristic peaks at 2 theta values of 6.3 degrees +/-0.2 degrees, 12.7 degrees +/-0.2 degrees and 15.9 degrees +/-0.2 degrees by using Cu-Kalpha radiation

2. Crystalline form of compound I mesylate according to claim 1, which is characterized by an X-ray powder diffraction pattern having characteristic peaks at least one of 2 Θ values of 7.9 ° ± 0.2 °, 19.2 ° ± 0.2 °, 19.9 ° ± 0.2 °, using Cu-ka radiation.

3. The crystalline form of compound I mesylate according to claim 2, which has an X-ray powder diffraction pattern with characteristic peaks at least one of 2 Θ values of 14.5 ° ± 0.2 °, 20.4 ° ± 0.2 °, using Cu-ka radiation.

4. The crystalline form of compound I mesylate according to claim 1, which is characterized by an X-ray powder diffraction pattern substantially as shown in figure 3.

5. A process for preparing a crystalline form of compound I mesylate according to claim 1, wherein the process comprises: and mixing the solid of the compound I, a mixed solvent of methanesulfonic acid, alcohol and water or a mixed solvent of ether and water, and stirring to obtain the crystal form of the mesylate of the compound I.

6. The process for preparing a crystalline form of compound I mesylate according to claim 5, wherein the alcoholic solvent is isopropanol; the ether solvent is tetrahydrofuran; the molar ratio of the compound I solid to the methanesulfonic acid is 0.9:1-1.1: 1; the volume ratio of the alcohols to the water or the ethers to the water in the mixed solvent is 9: 1.

7. A pharmaceutical composition comprising a therapeutically effective amount of a crystalline form of compound I mesylate according to claim 1, and a pharmaceutically acceptable excipient.

8. Use of the crystalline form of compound I mesylate salt, as claimed in claim 1, in the preparation of a ROCK2 inhibitor medicament.

9. Use of a crystalline form of compound I mesylate as claimed in claim 1, for the preparation of a medicament for the treatment of chronic graft versus host disease, systemic sclerosis, and idiopathic pulmonary fibrosis.