CN111848677B

CN111848677B - Crystal form of ALK kinase inhibitor compound, preparation method and application

Info

Publication number: CN111848677B
Application number: CN201910351966.2A
Authority: CN
Inventors: 吴豫生; 李钧; 郑茂林; 梁阿朋; 牛成山
Original assignee: Deuracor Therapeutic Inc
Current assignee: Deuracor Therapeutic Inc
Priority date: 2019-04-29
Filing date: 2019-04-29
Publication date: 2023-03-17
Anticipated expiration: 2039-04-29
Also published as: CN111848677A

Abstract

The invention belongs to the technical field of medical compounds, and particularly discloses multiple crystal forms of an ALK kinase inhibitor compound, a preparation method and a pharmaceutical application thereof. By integrating the thermodynamic stability relationship and the solid property evaluation result, the crystal form B provided by the invention is an optimal crystal form, has stable physicochemical properties, and is very suitable for application in the aspects of drug development and the like of next preparations. The crystal forms A, B, C, D, E, F, G, H, I, J, K and L provided by the invention can be used for preparing ALK inhibitor drugs, especially the crystal form B.

Description

Crystal form, preparation method and application of ALK kinase inhibitor compound

Technical Field

The invention relates to a crystal form of an ALK kinase inhibitor compound, a preparation method of the crystal form and application of the crystal form in the aspect of medicines, and belongs to the technical field of medicinal compounds.

Background

Molecular targeted therapy is the most promising therapeutic direction in the current tumor field, and the Epidermal Growth Factor Receptor (EGFR) and Anaplastic Lymphoma Kinase (ALK) are the most effective therapeutic targets for lung cancer. Related studies have shown that the most common genomic ALK abnormality in human tumors is a chromosomal rearrangement that results in the production of a fusion gene. Targeted therapies directed against ALK fusion proteins and constitutive activation units have achieved good efficacy in the treatment of a variety of malignancies. In various malignant tumors such as Anaplastic Large Cell Lymphoma (ALCL), inflammatory Myofibroblastoma (IMTs), neuroblastoma (NB), non-small cell lung cancer (NSCLC), and the like, ALK gene mutation or chromosomal translocation causes abnormal expression of ALK signaling pathway, thereby promoting tumor progression. The mutant and aberrant activity of ALK in various cancers has made it a drug target for the treatment of ALK-positive cancers.

Crizotinib (Crizotinib) developed by bufferi pharmaceutical co ltd, usa is the first ALK inhibitor introduced in the treatment of ALK-dependent NSCLC, is an oral small molecule multi-target tyrosine kinase inhibitor, and acts on ALK, c-MET and ROS1. Clinical verification shows that the malignant tumor size of the patient with the late gene mutant type non-small cell lung cancer (NSCLC) can be effectively reduced. However, crizotinib may exhibit the following side effects: visual disturbance and gastrointestinal side effects, the level of hepatic transaminase is increased in grade 3-4 in 16% of patients, and in addition, acquired drug resistance inevitably occurs in ALK positive patients after the sensitive period of crizotinib treatment in the initial stage, and researches show that the patients generally have drug resistance after 1-2 years of treatment. The treatment of the second generation ALK inhibitors such as LDK378, alectinib and AP26113 found in early researches at present improves the curative effect, but the second generation ALK inhibitors have the problem of great side effect.

Chinese patent application publication No. CN108689994A discloses a series of compounds useful as ALK kinase inhibitors, all of which have very good ALK inhibitory activity, higher than that of crizotinib and brigitinib, etc., which are currently on the market. In this patent application, a compound of formula (I) is disclosed. At present, no patent report about the crystal form of the compound is found. The crystal form of the compound shown in the formula (I) is screened and developed, and a stable and reliable crystal form substance is found to ensure the quality stability of the compound shown in the formula (I) so as to be better used for pharmacy and pharmaceutical preparations and clinical use in the future, so that the stability and controllability of the quality of the medicine, better dissolution and higher bioavailability are achieved, and the safety problems of side effects and the like caused by toxic impurities and the like generated due to the instability of the medicine are avoided, so that the method has important significance for later promotion.

Disclosure of Invention

The invention researches the crystal form of the compound shown in the formula (I), provides several crystal forms of the compound shown in the formula (I), provides a preparation method and also provides application of the compound in the aspect of medicaments.

Wherein, the invention provides a crystal form B of a compound shown as a formula (I),

the X-ray powder diffraction of the crystal form B measured by using Cu-Kalpha rays is as follows at the angle of 2 theta: diffraction peaks are present at 6.07 +/-0.2 degrees, 13.24 +/-0.2 degrees, 21.86 +/-0.2 degrees, 23.93 +/-0.2 degrees and 24.32 +/-0.2 degrees.

Further, form B of the compound of formula (i) has an X-ray powder diffraction at 2 Θ angle measured using Cu-K α radiation: diffraction peaks are also found at 9.38 + -0.2 deg., 17.37 + -0.2 deg., 18.34 + -0.2 deg., 19.64 + -0.2 deg., 20.10 + -0.2 deg., 22.23 + -0.2 deg., 23.39 + -0.2 deg., 26.61 + -0.2 deg., 27.65 + -0.2 deg., 29.82 + -0.2 deg., 30.52 + -0.2 deg., 31.64 + -0.2 deg., 33.65 + -0.2 deg., 35.72 + -0.2 deg., and 36.81 + -0.2 deg..

Further, the X-ray powder diffraction pattern of the crystal form B of the compound shown in the formula (I) measured by using Cu-K alpha rays is basically shown in figure 2.

Further, the differential scanning calorimetry analysis pattern of the crystal form B shows an endothermic peak at 204.7 ℃ (starting temperature).

Further, the form B is an anhydrous form.

The invention also provides a preparation method of the crystal form B of the compound shown in the formula (I), which comprises the following steps:

s1: preparation of Crystal form A

Adding a compound shown in a formula (I) into water, then adding dilute hydrochloric acid, stirring for dissolving, after completely dissolving, dropwise adding 0.2mol/L sodium hydroxide aqueous solution at 60 ℃ until the pH value of a system is 11-12, cooling to room temperature, filtering, and drying to obtain a crystal form A;

s2: preparation of form B

Suspending and stirring the crystal form A in isopropanol at room temperature to obtain a crystal form B;

or the crystal form A is subjected to gas-solid diffusion, gas-liquid diffusion, suspension stirring at 50 ℃, slow volatilization, addition of an anti-solvent, slow cooling or high polymer induction to obtain the crystal form B.

The crystal form B of the compound shown in the formula (I) provided by the invention has the following application.

The invention provides application of a crystal form B of a compound shown as a formula (I) in preparation of a medicine for inhibiting cell proliferation.

The invention provides application of a crystal form B of a compound shown as a formula (I) in preparation of a medicine for treating cancer.

The invention provides application of a crystal form B of a compound shown as a formula (I) in preparation of a medicine for inhibiting anaplastic lymphoma kinase.

The invention provides application of a crystal form B of a compound shown as a formula (I) in preparation of a medicine for treating non-small cell lung cancer.

The invention also provides a pharmaceutical composition containing the crystal form B of the compound shown in the formula (I).

The crystal form is the solid matter state existing in the medicine, the medicine crystal form research is the research on the basic state of the medicine, and the medicine crystal form solid matter more suitable for treating diseases can be possibly searched only if the crystal form state of the chemical medicine is fully and comprehensively known. The crystal form of the medicine can influence the physicochemical property of the medicine and directly influence the basis of the clinical application of the medicine for treating diseases. Different crystal forms of the same drug may have significant differences in appearance, solubility, melting point, dissolution rate, bioavailability and the like, thereby affecting the stability, bioavailability and therapeutic effect of the drug. Therefore, the research on the stable crystal form of the compound has important significance.

The active ingredients of the same drug generally exist in two or more crystal forms, referred to as drug polymorphs. Different crystal forms have different respective solubilities and dissolution rates, and the clinical treatment effect of the medicine is influenced by causing the change of in vivo bioavailability. Different crystal forms of the drug may affect the dissolution and absorption of the drug in vivo, thereby affecting the bioavailability, clinical efficacy and safety of the drug. Meanwhile, the stability of the drug crystal form is also very important. In order to improve the bioavailability of the drug, reduce toxicity and improve the therapeutic effect, more attention is paid to the stability of the drug crystal form. The crystal form with good stability can ensure the physical and chemical stability of the medicament form in the preparation and storage processes, maintain the good solubility and bioavailability of the medicament form and ensure the equivalence among each batch of medicaments. The same drug often has a plurality of crystal forms, and the crystal form which has better treatment effect and is most suitable for clinic is called as the dominant drug crystal form at present.

The invention screens the crystal forms of the compound shown in the formula (I) and finds out different crystal forms of the raw material medicines as much as possible. The screened crystal form is identified by means of powder X-ray diffraction analysis (XRPD), differential Scanning Calorimetry (DSC), thermogravimetric analysis (TGA), hydrogen spectrum liquid nuclear magnetism (1H NMR), high Performance Liquid Chromatography (HPLC) and the like, and further through research and investigation of thermodynamic stability and hygroscopicity, the crystal form B of the dominant drug is determined, so that a reference basis is provided for crystal form selection of subsequent pharmacokinetic experiments and zoology experiments.

Through further pharmacokinetic research experiments, the compound crystal form B shown in the formula (I) provided by the invention is easy to absorb in a living body and has good stability, good blood concentration distribution in the living body and high bioavailability. Meanwhile, the physical stability of the compound preparation is good, so that the compound preparation is very beneficial to the quality control of the medicine in the later preparation development and has longer quality guarantee period, thereby leading the medicine to achieve better curative effect.

Drawings

FIG. 1 is an X-ray powder diffraction pattern of the crystal form A provided by the invention measured by Cu-Ka rays;

FIG. 2 is an X-ray powder diffraction pattern of the crystal form B provided by the invention measured by Cu-Ka ray;

FIG. 3 is an X-ray powder diffraction pattern of form C provided by the present invention as measured using Cu-Ka radiation;

FIG. 4 is an X-ray powder diffraction pattern of form D provided by the present invention measured using Cu-Ka radiation;

FIG. 5 is an X-ray powder diffraction pattern of form E provided by the present invention as measured using Cu-Ka radiation;

FIG. 6 is an X-ray powder diffraction pattern of the crystal form F provided by the invention measured by Cu-Ka ray;

FIG. 7 is an X-ray powder diffraction pattern of form G provided by the present invention as measured using Cu-Ka radiation;

FIG. 8 is an X-ray powder diffraction pattern of form H provided by the present invention as measured using Cu-Ka radiation;

FIG. 9 is an X-ray powder diffraction pattern of the crystal form I provided by the invention measured by Cu-Ka ray;

FIG. 10 is an X-ray powder diffraction pattern of form J provided by the present invention as measured using Cu-Ka radiation;

FIG. 11 is an X-ray powder diffraction pattern of form K provided by the present invention as measured using Cu-Ka radiation;

FIG. 12 is an X-ray powder diffraction pattern of form L provided by the present invention as measured using Cu-Ka radiation;

FIG. 13 is a graph of the transformation relationship between various crystalline forms provided by the present invention;

figure 14 is a TGA/DSC profile of form a provided by the present invention;

figure 15 is a TGA/DSC profile of form B provided by the present invention;

figure 16 is a TGA/DSC profile of form C provided by the present invention;

FIG. 17 is a drawing of form C provided by the present invention ¹ An H-NMR spectrum;

figure 18 is an overlay of X-ray powder diffraction patterns provided by the present invention for form C, before and after heating;

FIG. 19 is an overlay of X-ray powder diffraction patterns of crystalline form C provided by the present invention which is prepared repeatedly by slow evaporation in different solvent systems;

figure 20 is a TGA/DSC profile of form C provided by the present invention for slow evaporation in EtOH solvent;

FIG. 21 is a schematic representation of a preferred embodiment of the present inventionSlowly evaporating repeatedly prepared crystalline form C in EtOH solvent ¹ An H-NMR spectrum;

figure 22 is an X-ray powder diffraction overlay pattern before and after heating of crystalline form C provided in accordance with the present invention, which is slowly volatilized from EtOH solvent and repeatedly prepared;

figure 23 is a TGA/DSC profile of form D provided by the present invention;

FIG. 24 is an X-ray powder diffraction stacking pattern of crystalline form D provided by the present invention before and after heating to 195 ℃;

figure 25 is a DVS profile of form D provided by the present invention;

figure 26 is an X-ray powder diffraction overlay pattern before and after DVS for form D provided by the present invention;

figure 27 is a TGA/DSC profile of form E provided by the present invention;

FIG. 28 is an X-ray powder diffraction stacking pattern of form E provided herein before and after heating to 160 ℃;

FIG. 29 shows a crystal form E provided by the present invention ¹ An H-NMR spectrum;

figure 30 is a TGA/DSC profile of form F provided by the present invention;

FIG. 31 is an X-ray powder diffraction stacking pattern of form F provided by the present invention before and after heating to 120 ℃;

FIG. 32 shows a crystalline form F provided by the present invention ¹ An H-NMR spectrum;

FIG. 33 is a superimposed X-ray powder diffraction pattern of a selected sample of form G (reference in the figure) and a repeatedly prepared sample of form G after drying;

figure 34 is a TGA/DSC profile of form G provided by the present invention;

FIG. 35 is an X-ray powder diffraction overlay pattern of form H provided herein before and after drying;

figure 36 is a TGA/DSC profile of form I provided by the present invention;

figure 37 is a TGA/DSC profile of form J provided herein;

figure 38 is a TGA/DSC profile of form K provided by the present invention;

FIG. 39 is an X-ray powder diffraction overlay spectrum of form K provided by the present invention after heating to 140 ℃ and 180 ℃;

figure 40 is a TGA profile of crystalline form K provided by the present invention after heating to 140 ℃;

FIG. 41 shows a crystal form K provided by the present invention ¹ An H-NMR spectrum;

fig. 42 is an X-ray powder diffraction overlay of form L provided herein and repeatedly prepared form L;

figure 43 is a TGA/DSC profile of form L provided by the present invention;

figure 44 is an X-ray powder diffraction contrast plot of an anhydrous crystalline form suspension competition sample provided by the present invention;

figure 45 is a first graph of X-ray powder diffraction contrast of a competing suspension of forms B and F provided by the present invention to give a solid;

FIG. 46 is a second graph of X-ray powder diffraction contrast of a solid competing suspension of forms B and F provided by the present invention;

FIG. 47 is an X-ray powder diffraction overlay plot of stability experiment for form B provided herein;

figure 48 is a DVS profile of form B provided by the present invention;

figure 49 is an X-ray powder diffraction overlay pattern of form B provided by the present invention before and after DVS testing;

FIG. 50 is a PLM spectrum of form B provided herein;

figure 51 is a pharmacokinetic study profile of form a and form B provided herein;

FIG. 52 is an X-ray powder diffraction pattern of a compound represented by formula (I) synthesized according to the production method disclosed in the present invention with reference to publication No. CN108689994A, measured by Cu-Ka radiation.

Detailed Description

The technical solution of the present invention will be explained in detail below.

Referring first to the preparation method disclosed in patent application publication No. CN108689994A, the compound represented by formula (I) was synthesized as a pale yellow solid, and its X-ray powder diffraction pattern measured using Cu-K.alpha.radiation is shown in FIG. 52.

In subsequent studies, it was found that the compound prepared by the preparation method disclosed in patent application publication No. CN108689994A is liable to absorb moisture and deteriorate, and the compound is poor in stability and liable to generate new impurities. Therefore, there is a need to develop stable crystalline forms of the compound for preclinical and clinical studies.

Subsequently, we carried out a number of experiments for the development of pharmaceutically stable crystalline forms of the compound of formula (i).

First, we have tested the solubility of the compound of formula (i) in different solvents (see table 1 for details), and we have found that the solubility of this compound in various solvents is very different, and that the solubility in the same type of solvent is also very different. We have also found that the solubility of the compounds of formula (I) in non-alcoholic solvents is very close to that in alcoholic solvents.

Then, screening the polymorphic crystal of the compound shown in the structural formula (I) by adopting a method comprising gas-solid diffusion, gas-liquid diffusion, suspension stirring at room temperature, suspension stirring at 50 ℃, slow volatilization, anti-solvent addition, slow temperature reduction, high polymer induction, grinding, suspension crystallization, volatilization crystallization and temperature reduction crystallization, wherein 100 polymorphic crystal screening tests are totally set, and specific test methods and results are summarized in a table 2. After a number of failures and then a summary of experience from the failures, we have developed several polymorphic forms of the compound of formula (i). In the next study, form B was found to be very stable by analysis such as TGA and DSC, as well as stability experiments. Finally, the crystal form substance is determined to be used as a stable crystal form for continuously promoting the research of the medicine, and is used for subsequent evaluation and clinical development.

The specific crystal form study process is described in the following section.

Firstly, taking a prepared compound shown in a formula (I) as a raw material, adding the compound shown in the formula (I) into water, then adding dilute hydrochloric acid, stirring and dissolving, then dropwise adding 0.2mol/L sodium hydroxide aqueous solution at 60 ℃, separating out solids, preserving heat, filtering, and drying to obtain a crystal form substance, namely a crystal form A.

Thereafter, a solubility test was carried out to test the crude solubility of form a of the compound of structural formula (i) in 20 common solvents at room temperature. In the test, 2 mg of solid sample is weighed into a 3 ml vial, and the corresponding solvent is added gradually (50/50/200/700/1000 μ l in sequence) and shaken until the solid is clear. If the sample is not clear after the solvent is added to 2 ml, no further solvent is added. The calculated coarse solubility range, based on the solid sample mass, the volume of solvent added and the observed dissolution phenomenon, is shown to guide the design of the screening assay. The crude solubility at room temperature of form a of the compound of formula (i) is shown in table 1.

Table 1 table of rough solubility of form a at room temperature

1. Polymorphism screening test

The screening method for the polymorphic crystal of the compound shown in the structural formula (I) comprises the steps of gas-solid diffusion, gas-liquid diffusion, suspension stirring at room temperature, suspension stirring at 50 ℃, slow volatilization, anti-solvent addition, slow cooling, high polymer induction, grinding, suspension crystallization, volatilization crystallization and cooling crystallization, 100 polymorphic crystal screening tests are set in total, and specific test methods and results are summarized in a table 2.

TABLE 2 summary of polymorphic form screening assays

Method	Number of experiments	Results
			Gas-solid diffusion	13	Forms A, B, E, F and H
Gas-liquid diffusion	10	Crystal forms A, B and G
			Suspending and stirring at room temperature	20	Forms A, B, C and J
Suspension stirring at 50 DEG C	14	Crystal forms A and B
			Slowly volatilize	12	Forms B, C, E, F and J
Addition of anti-solvent	12	Crystal forms A, B, C and D
			Slowly cool down	10	Crystal forms A, B and I
High Polymer Induction	6	Crystal forms A, B and C
			Grinding	3	Amorphous form
In total	100	Crystal forms A to J and amorphous forms

1.1 gas-solid diffusion

A total of 13 gas-solid diffusion tests were set up with different solvents. About 20 mg of compound form a of formula (i) was weighed into a 3 ml vial, about 4 ml of solvent was added to a 20 ml vial, and after the 3 ml vial was opened into the 20 ml vial, the 20 ml vial was sealed. After standing at room temperature for 10 days, the solid was collected and tested for XRPD. The test results are shown in table 3, and crystal forms a, B, E, F and H were obtained in the gas-solid permeation test.

TABLE 3 summary of gas-solid diffusion test

* : after 10 days, the sample is in a solution state and is evaporated and dried at room temperature to obtain the sample.

1.2 gas-liquid diffusion

In a 3 ml vial, about 20 mg per part of the compound form a represented by the structural formula (i) was weighed and dissolved in 0.1 to 0.5 ml of a good solvent (see table 4), about 3 ml of an anti-solvent was added to another 20 ml vial, the 3 ml vial containing the clear solution was placed in the 20 ml vial with the opening, and then the 20 ml vial was sealed and left to stand at room temperature. When solid evolution was observed, the solid was collected and tested for XRPD. The test results are shown in table 4, and the gas-liquid diffusion test yielded crystal forms a, B, and G.

TABLE 4 summary of gas-liquid diffusion test

1.3 Room temperature suspension stirring

About 15 mg per part of the compound form a of the formula (i) was weighed into an HPLC vial, 0.3 to 0.4 ml of the solvents listed in table 5 were added, respectively, and after the resulting suspension was magnetically stirred (about 800 rpm) at room temperature for about 4days, the solid was centrifuged and subjected to XRPD test. The test results are shown in table 5, and crystal forms a, B, C and J were obtained by room temperature suspension stirring test.

TABLE 5 summary of room temperature suspension stirring test

Test number	Solvent (V/V)	As a result, the
			819805-06-A1	Acetic acid isopropyl ester	Crystal form B
819805-06-A2	Isopropanol (I-propanol)	Crystal form B
			819805-06-A3	Methyl isobutyl ketone	Crystal form B
819805-06-A4	Ethyl acetate	Crystal form B
			819805-06-A5	Methyl tert-butyl ether	Crystal form B
819805-06-A6	Acetonitrile	Crystal form B
			819805-06-A7	N-heptane	Crystal form B
819805-06-A8	Toluene	Crystal form B
			819805-06-A9	Water (W)	Crystal form A
819805-06-A10	Dichloromethane/n-heptane (1	Crystal form B
			819805-06-A11	Methanol/methyl tert-butyl ether (1	Crystal form C
819805-06-A12	Tetrahydrofuran/isopropyl acetate (1	Crystal form B
			819805-06-A13	Ethanol/water (1	Crystal form A
819805-06-A14	Chloroform/isopropyl acetate (1	Crystal form B
			819805-06-A15	Tetrahydrofuran/water (1	Crystal form A
819805-06-A16	Methylene chloride/methyl tert-butyl ether (1	Crystal form B
			819805-06-A17	Isopropyl alcohol/water aw 0.2	Crystal form B
819805-06-A18	Isopropyl alcohol/water aw 0.4	Crystal form B
			819805-06-A19	isopropanol/Water aw 0.6	Crystal form B
819805-06-A20	isopropanol/Water aw 0.8	Form J

* : suspending and stirring for 3 days at room temperature, dissolving the sample, transferring to 5 ℃, continuously stirring, still dissolving the sample, and performing vacuum drying to obtain a solid sample.

1.4 Suspending and stirring at 50 DEG C

About 15 mg of each of the compound form a of the formula (i) was weighed into an HPLC vial, 0.3 ml of each of the solvents listed in table 6 was added, and after the resulting suspension was placed under magnetic stirring (about 800 rpm) at 50 ℃ for about 4days, the solid was separated by centrifugation and subjected to XRPD test. The test results are shown in table 6, and the crystal forms a and B are obtained by suspension stirring test at 50 ℃.

TABLE 6 summary of 50 ℃ suspension stirring test

1.5 slow volatilization

A total of 12 slow evaporation tests were set up using different solvent systems. Respectively weighing 15 mg of the compound crystal form A shown in the structural formula (I) into 3 ml small bottles, respectively adding 0.2-2.0 ml of the solvent shown in the table 7, if the solvent is not clarified after 2.0 ml of the solvent is added, filtering the solution (a PTFE filter membrane with the pore diameter of 0.45 micrometer) to obtain a clarified solution, sealing the small bottles filled with the clarified solution by using a sealing membrane, pricking 5 small holes on the bottle, and standing at room temperature for slow volatilization. When the solvent was completely evaporated, the resulting solid was collected and tested for XRPD. The results are shown in table 7, and form B, C, E, F and J were obtained in the slow volatilization test.

TABLE 7 summary of slow volatilization tests

1.6 anti-solvent addition

Weighing about 15 mg of each part of the compound crystal form A shown in the structural formula (I) and adding the compound crystal form A into a 20 ml small bottle, dissolving the compound crystal form A with 0.2-2.0 ml of a good solvent (shown in table 8), adding the anti-solvent shown in table 8 into the clear solution, stirring while dripping until solid is separated out, and stopping when no solid is separated out after adding about 10ml of the anti-solvent. Solids were separated by centrifugation and XRPD tested, with the results shown in table 8, and forms a, B, C and D were obtained in the anti-solvent addition test.

TABLE 8 antisolvent addition test summary

^* :10 ml of solvent is added dropwise, then clarification is carried out, and solid is precipitated after the stirring time is prolonged.

^# : and (3) adding 10ml of an anti-solvent dropwise, clarifying, transferring to 5 ℃, stirring, still not precipitating a solid, and transferring to room temperature to volatilize to obtain the solid.

A: the samples were layered and the lower layer was evaporated at room temperature.

1.7 Slow Cooling

Weighing about 15 mg of compound crystal form A shown in the structural formula (I) in each part, adding 1.2-2.0 ml of solvent shown in table 9 into a 3 ml small bottle, stirring at 50 ℃ for about 2 hours, filtering (a PTFE filter membrane with the pore diameter of 0.45 micron) to obtain a filtrate, placing the obtained filtrate into a biochemical incubator, cooling from 50 ℃ to 5 ℃ at the cooling speed of 0.1 ℃/minute, collecting precipitated solid, and carrying out XRPD test. The test results are shown in Table 9. And performing slow cooling test to obtain crystal forms A, B and I.

TABLE 9 summary of slow cooling test

^* : no solid is separated out after cooling, no solid is separated out after transferring to-20 ℃, and the solid is obtained after vacuum drying.

1.8 high Polymer Induction

Weighing about 15 mg of each part of the compound crystal form A shown in the structural formula (I) and dissolving the compound crystal form A in 0.2-2.0 ml of the solvent in the table 10, if the solvent is added to exceed 2.0 ml, filtering (a PTFE filter membrane with the pore diameter of 0.45 micron), taking the filtrate, transferring the filtrate into a 3 ml small bottle filled with about 2 mg of mixed high polymer, sealing the small bottle filled with the filtrate by using a sealing membrane, pricking 3-5 small holes on the small bottle, standing at room temperature for slow volatilization, collecting the obtained solid, and carrying out XRPD test. The test results are shown in table 10, and crystal forms a, B and C were obtained in the polymer induced crystallization test.

TABLE 10 summary of Polymer Induction test

Wherein, the mixed high polymer A: polyvinylpyrrolidone, polyvinyl alcohol, polyvinyl chloride, polyvinyl acetate, hydroxypropyl methylcellulose and methylcellulose (mixed by equal mass);

mixing a high polymer B: polycaprolactone, polyethylene glycol, polymethyl methacrylate, sodium alginate and hydroxyethyl cellulose (mixed by equal mass).

2. Analysis of crystal form

The resulting solid sample was analyzed using various detection analysis methods, such as powder X-ray diffraction (XRPD), differential Scanning Calorimetry (DSC), and thermogravimetric analysis (TGA), dynamic moisture sorption (DVS).

2.1 powder X-ray diffraction (XRPD)

XRPD patterns were collected on a PANALYtic Empyrean and X' Pert3 ray powder diffraction Analyzer with the scanning parameters shown in Table 11.

TABLE 11XRPD test parameters

2.2 thermogravimetric analysis (TGA) and Differential Scanning Calorimetry (DSC)

TGA and DSC plots were taken on a TA Q500/Q5000/Discovery 5500 thermogravimetric analyzer and a TA Q200/Q2000/Discovery 2500 differential scanning calorimeter, respectively, with the test parameters listed in Table 12.

TABLE 12TGA and DSC test parameters

2.3 dynamic moisture sorption (DVS)

Dynamic water sorption (DVS) curves were collected on DVS Intrinsic of SMS (Surface Measurement Systems). At a relative humidity of 25 deg.C using LiCl, mg (NO) ₃ ) ₂ And deliquescence point correction of KCl. The DVS test parameters are listed in table 13.

TABLE 13DVS test parameters

2.4 liquid Nuclear magnetic resonance of Hydrogen Spectroscopy: ( ¹ H Solution NMR)

Collecting hydrogen spectrum liquid-state nuclear magnetic spectrum on Bruker 400M nuclear magnetic resonance instrument, DMSO-d ₆ As a solvent.

2.5 polarizing microscope (PLM)

The polarization microscopy data were collected by an Axio scope.a1 microscope from Carl Zeiss German.

2.6 High Performance Liquid Chromatography (HPLC)

The purity of the samples in the test was measured by Agilent 1260 high performance liquid chromatography with the analytical conditions as given in Table 14.

TABLE 14 HPLC TEST CONDITIONS FOR PURITY TESTS

3. Polymorph characterization and identification

3.1 form A

The preparation method of the crystal form A comprises the following specific steps: adding a compound shown in a formula (I) into water, adding 1M/HCl, stirring for dissolving, placing the mixture at 60 ℃ for stirring, dropwise adding 0.2M sodium hydroxide aqueous solution at the temperature, dropping the solution at a speed of about 1 drop/10 seconds until the pH value is = 11-12, cooling to room temperature, filtering, and drying to obtain the crystal form A, wherein the concentration is 33.3 mg/mL.

XRPD results for form a are shown in figure 1. TGA and DSC results are shown in figure 14, with a 1.4% weight loss for the form a sample heated to 150 ℃, and an endothermic melting peak in DSC only at 198.6 ℃ (onset temperature). And (4) integrating the TGA data and the DSC data, and judging the crystal form A to be an anhydrous crystal form. HPLC results showed the purity of form a sample to be 98.9% (area normalization).

As can be seen from fig. 1, X-ray powder diffraction peak data of form a is shown in table 15.

Watch 15

3.2 form B

The crystal form B sample is obtained by suspending and stirring the crystal form A sample in IPA solvent at room temperature, and XRPD is shown in figure 2. For example, the preparation method of the crystal form B can be as follows: and suspending and stirring the crystal form A sample in isopropanol at room temperature. In addition, the crystal form B can be obtained under the conditions of gas-solid diffusion, gas-liquid diffusion, suspension stirring at 50 ℃, slow volatilization, addition of an anti-solvent, slow temperature reduction and high polymer induction. TGA and DSC results are shown in figure 15, which indicate that the form B sample (819805-14-B) heated to 150 ℃ had a 1.6% weight loss; DSC results showed that the sample had an endothermic peak only at 204.7 ℃ (onset temperature), presumably due to melting of form B. HPLC results indicated that the purity of the form B sample was 99.7% (area normalization). The results suggest that form B is an anhydrous form.

As can be seen from fig. 2, X-ray powder diffraction peak data of form B is shown in table 16.

TABLE 16

3.3 form C

Form C samples (819805-14-C1) were obtained by slow evaporation in MeOH solvent. Form C can also be obtained by stirring in MeOH/MTBE (1, 4,v/v) solvent at room temperature, slow evaporation in EtOH solvent, addition of EtOH/n-heptane system anti-solvent, and the like.

Slow evaporation in MeOH gave three batches of form C samples (819805-09-A8, 819805-14-C1, and 819805-19-C-dry4 days) with XRPD results as shown in FIG. 3. A sample of form C (819805-19-C-dry 4 days) was subjected to TGA, DSC and ¹ the results of H-NMR are shown in FIGS. 16 and 17. The TGA results show that a sample of form C (819805-19-C-dry 4 days) had a 3.1% weight loss when heated to 100 ℃ and a 1.8% weight loss when heated further to 140 ℃. The DSC result showed that the sample had endothermic peaks at 99.8 ℃, 120.6 ℃, 188.5 ℃ and 202.9 ℃ (peak temperature) and exothermic peaks at 190.8 ℃ (peak temperature); the nuclear magnetic results show that a sample of form C (819805-19-C-dry 4 days) has a MeOH solvent peak, the MeOH/API molar ratio is 0.37, and the corresponding weight loss is 1.9%, which is substantially consistent with the second weight loss in TGA. XRPD (X-ray diffraction pattern) is tested after the crystal form C sample (819805-14-C1) is heated to 140 ℃ and 195 ℃ respectively, the result is shown in figure 18, crystal transformation occurs after the crystal form C sample (819805-14-C1) is heated to 140 ℃, and a new crystal formIs named as crystal form L; heating to 195 deg.C and crystallizing to form crystal form B. The form C sample (819805-19-C-dry 4 days) was presumed to be MeOH solvate.

The preparation of the form C sample (819805-14-C2) was repeated with slow evaporation in EtOH and the XRPD pattern is shown in fig. 19. A sample of form C (819805-14-C2) was subjected to TGA, DSC and ¹ the results of H-NMR are shown in FIGS. 20 and 21. The TGA results showed that there was a 9.7% weight loss of the form C sample (819805-14-C2) when heated to 130 ℃; the DSC result showed that the sample had endothermic peaks at 95.1 ℃, 116.0 ℃, 181.2 ℃ and 204.9 ℃ (peak temperature) and exothermic peaks at 188.9 ℃ (peak temperature); the nuclear magnetic results show that the crystal form C sample (819805-14-C2) has an EtOH solvent peak, the molar ratio of EtOH to API is 0.8, and the corresponding weight loss is 5.6%. After the crystal form C sample (819805-14-C2) is heated to 130 ℃ and 195 ℃ respectively, XRPD is tested, the result is shown in figure 22, crystal transformation occurs after the crystal form C sample (819805-14-C2) is heated to 130 ℃, and the new crystal form is a crystal form L; heating to 195 deg.C and crystallizing to form crystal form B. The form C sample (819805-14-C2) was presumed to be an EtOH solvate.

And (3) integrating the characterization results of the two types of crystal form C samples, and judging that the crystal form C is a solvate, wherein the solvent can be MeOH or EtOH.

As can be seen from fig. 3, X-ray powder diffraction peak data of form C is shown in table 17.

TABLE 17

2θ(°)	Relative Strength (%)
		6.71	100.00
8.90	22.04
		9.68	1.66
10.97	5.05
		13.42	61.65
14.05	6.17
		15.09	11.44
17.48	5.54
		17.84	8.03
18.73	54.66
		20.18	55.51
21.84	13.47
		22.16	43.81
23.83	2.47
		25.10	9.41
28.38	2.34
		28.80	5.01
33.94	2.08

3.4 form D

Three batches of form D samples (819805-09-A5, 819805-14-D and 819805-18-D) were obtained by DCM/MTBE system anti-solvent addition with XRPD results as shown in figure 4. TGA and DSC results for the form D sample (819805-09-A5) are shown in fig. 23, which indicate a 2.9% weight loss of the form D sample (819805-09-A5) when heated to 200 ℃; the DSC results showed that the sample had an endothermic peak at 189.3 ℃ (peak temperature) and an exothermic peak at 190.5 ℃ (peak temperature), presumably due to sample transcrystallization; the presence of an endothermic peak at 203.4 deg.C (starting temperature) is presumed to be caused by melting of the recrystallized sample.

After heating the form D sample (819805-14-D) to 195 ℃, the sample transforms to form B, the XRPD overlay is shown in fig. 24. The DVS profile of the sample of form D (819805-18-D) is shown in FIG. 25, the DVS results indicate a weight gain of about 0.5% of form D from 0% RH-80% RH, indicating that form D is slightly hygroscopic; the crystal form of form D before and after DVS was unchanged, and the XRPD overlay before and after DVS is shown in fig. 26. The result is combined to infer that the crystal form D is an anhydrous crystal form.

As can be seen from fig. 4, the X-ray powder diffraction peak data of form D is shown in table 18.

Watch 18

2θ(°)	Relative Strength (%)
		6.64	100.00
8.75	15.79
		9.02	11.67
10.60	16.47
		11.36	5.92
13.28	78.47
		14.62	4.76
15.38	7.30
		16.16	8.07
16.93	6.05
		17.51	7.84
17.97	3.08
		19.17	3.53
19.97	45.69
		20.42	31.63
21.12	17.94
		22.39	3.49
22.82	4.70
		23.32	5.19
24.14	6.12
		26.73	6.76
28.62	2.05

3.5 form E

Passing of the sample of form E (819805-14-E) through CHCl ₃ The solution was slowly evaporated and the XRPD results are shown in figure 5. TGA and DSC results are shown in FIG. 27, which indicate that form E (819805-14-E) has a weight loss of 12.1% when heated to 150 ℃; the DSC results showed that the sample had an endothermic peak at 110.6 ℃, 120.2 ℃ (peak temperature), and the presence of an endothermic peak at 203.3 ℃ (onset temperature) is presumed to be caused by melting of the sample. XRPD measurement of form E (819805-14-E) after heating to 160 ℃ revealed that form E crystallized to form B, with the XRPD overlay before and after heating shown in figure 28. Crystal form E (81)9805-14-E) ¹ The H-NMR is shown in FIG. 29, and the nuclear magnetic results show that CHCl is present in the sample of the crystal form E (819805-14-E) ₃ Wherein the molar ratio of chloroform to API was 0.85, corresponding to a weight loss of 14.0%. By combining the TGA/DSC and nuclear magnetic results, the crystal form E can be judged to be CHCl ₃ A solvate of (1).

As can be seen from FIG. 5, the X-ray powder diffraction peak data of form E is shown in Table 19.

Watch 19

2θ(°)	Relative Strength (%)
		12.48	20.50
15.77	26.66
		17.57	13.89
18.14	33.93
		18.59	100.00
19.62	25.92
		19.91	37.54
20.57	24.84
		20.91	67.18
21.57	23.40
		22.16	31.10
23.29	50.09
		25.28	16.41
25.99	15.92
		27.25	10.80
29.09	22.33
		30.01	12.32
30.82	7.49
		32.40	49.07
35.89	3.11

3.6 form F

Form F samples (819805-14-F) were obtained by slow evaporation of DCM solution and XRPD results are shown in fig. 6. TGA and DSC results are shown in figure 30, which indicate that there is a 5.5% weight loss of the crystalline form F sample (819805-14-F) when heated to 120 ℃; DSC results showed that the sample had an endothermic peak at 92.9 ℃ (peak temperature) presumably due to dehydration/solvent; the presence of an endothermic peak at 201.8 ℃ (onset temperature) is presumed to be caused by melting of the sample; XRPD was measured after heating the form F sample (819805-14-F) to 120 ℃ and the result was transformed to form B, which is shown in figure 31 for the XRPD overlay before and after heating. Of form F (819805-14-F) ¹ The H-NMR is shown in figure 32, and the nuclear magnetic results show that no obvious DCM solvent peak is observed in the crystal form F (819805-14-F). And (4) combining the characterization results, and judging that the crystal form F (819805-14-F) is a hydrate (theoretical weight loss of a dihydrate is 5.5%).

As can be seen from fig. 6, X-ray powder diffraction peak data of form F is shown in table 20.

Watch 20

2θ(°)	Relative Strength (%)
		6.75	100.00
9.14	30.34
		13.11	28.30
13.47	50.12
		14.63	22.61
18.34	34.31
		19.99	47.73
20.79	29.12
		23.28	59.18
26.12	11.18
		27.99	4.71

3.7 form G

Form G samples (819805-05-A4) were obtained by THF/n-heptane system antisolvent addition with XRPD results as shown in fig. 7. A repeatedly prepared sample of form G (819805-14-G) dried at room temperature to give form K, with the XRPD overlay shown in figure 33. TGA and DSC plots of form G sample (819805-05-A4) are shown in FIG. 34, and the TGA results indicate that the form G sample (819805-05-A4) has 5.1% weight loss when heated to 170 ℃; the DSC results showed exothermic peaks at 124.8 ℃ and 173.3 ℃ (peak temperature) for the sample; the presence of an endothermic peak at 200.3 deg.C (starting temperature) is presumed to be caused by melting of the sample. Since the crystal form G prepared repeatedly is transformed into the crystal form K after being dried at room temperature, further identification cannot be carried out.

As can be seen from fig. 7, X-ray powder diffraction peak data of form G is shown in table 21.

TABLE 21

2θ(°)	Relative Strength (%)
		4.97	17.00
6.27	100.00
		6.63	15.80
8.21	6.97
		9.99	10.64
11.43	18.80
		11.87	14.01
12.44	16.75
		12.97	14.78
13.32	9.42
		14.79	20.13
15.25	6.62
		16.15	12.78
17.57	10.00
		18.53	24.43
19.17	39.95
		19.44	36.00
19.79	27.75
		20.40	20.21
21.07	31.49
		22.06	26.81
24.28	11.50
		24.80	10.66
26.87	6.11
		28.71	3.27

3.8 form H

Form H samples (819805-04-a 10) were obtained by gas-solid diffusion of form a samples in EtOAc system, XRPD results are shown in figure 8. The form H sample was transformed to form B after evaporation at room temperature and the XRPD pattern is shown in figure 35. And repeatedly preparing the crystal form H by gas-solid diffusion of the crystal form A sample in an EtOAc system to obtain the crystal form A. Further characterization was not performed since form H was transcrystallized after drying and the repeated preparation was unsuccessful.

As can be seen from fig. 8, the X-ray powder diffraction peak data of form H is shown in table 22.

TABLE 22

3.9 form I

Form I samples (819805-10-A7) were obtained by slow cooling in a MeOH/MTBE (1,v/v) system followed by vacuum drying, with XRPD results as shown in figure 9. TGA and DSC results are shown in FIG. 36, which shows that there is a 5.7% weight loss of the form I sample (819805-10-A7) when heated to 190 ℃; DSC results showed that the sample had two endothermic peaks at 199.3 ℃ (peak temperature) and 202.7 ℃ (onset temperature). Repeated preparation of form I all yielded form B, the methods and results of which are summarized in table 23. Since the repeated preparation of form I was unsuccessful, no identification was performed.

TABLE 23 summary of repeated preparations of form I

As can be seen from fig. 9, X-ray powder diffraction peak data of form I is shown in table 24.

TABLE 24

2θ(°)	Relative Strength (%)
		5.84	54.87
6.07	13.69
		7.08	4.78
7.66	100.00
		9.52	3.25
11.65	32.07
		14.21	5.01
15.38	5.31
		17.04	11.79
17.60	6.44
		19.04	18.64
19.59	5.19
		20.38	11.41
22.46	24.81
		23.05	27.39
23.38	22.68
		24.35	4.06

3.10 form J

Form J samples (819805-08-A9) were obtained by slow evaporation from 2-MeTHF solutions with XRPD results as shown in FIG. 10. TGA and DSC results are shown in figure 37, which shows that a sample of form J (819805-08-A9) heated to 210 ℃ had a 4.1% weight loss; the DSC results show that the presence of an endothermic peak at 202.5 ℃ (onset temperature) of the sample is presumed to be caused by melting of the sample. Repeated preparations of form J all yielded form B, and the results are summarized in table 25. Since the repeated preparation of form J was unsuccessful, no identification was performed.

TABLE 25 summary of repeat preparations of form J

As can be seen from fig. 10, X-ray powder diffraction peak data of form J is shown in table 26.

Watch 26

2θ(°)	Relative Strength (%)
		13.08	41.74
17.12	47.93
		18.50	52.35
19.32	87.19
		19.81	100.00
21.53	61.63
		23.12	20.51
23.62	31.58
		27.48	52.89

3.11 form K

The crystal form K sample (819805-14-G _ dry) is obtained by drying the crystal form G sample (819805-14-G) at room temperature, and the XRPD result of the crystal form K is shown in figure 11. TGA/DSC results are shown in figure 38, which shows that a sample of form K (819805-14-G _ dry) heated to 180 ℃ had a 4.9% weight loss; the DSC result shows that the sample has an endothermic peak at 65.4 ℃ (peak temperature) which is supposed to remove the water or solvent adsorbed on the surface of the sample, and has an exothermic peak at 160.3 ℃ (peak temperature) which is supposed to be caused by the crystal transformation of the sample; the presence of an endothermic peak at 201.9 ℃ (starting temperature) is presumed to be caused by melting of the sample; the XRPD is measured after the crystal form K sample (819805-14-G _ dry) is heated to 140 ℃ and 180 ℃, the overlay is shown in figure 39, and the result shows that the crystal form K sample (819805-14-G _ dry) is still the crystal form K after being heated to 140 ℃, and is converted into the crystal form B after being heated to 195 ℃; TGA results for the sample of form K (819805-14-G _ dry) after heating to 140 ℃ are shown in fig. 40, showing a 2.4% weight loss upon heating to 180 ℃, significantly less than before heating (4.9%), presumably due to removal of surface moisture after heating; of form K (819805-14-G _ dry) ¹ The H-NMR is shown in figure 41, and the nuclear magnetic results show that no obvious THF solvent peak appears in the sample of the crystal form K (819805-14-G _ dry). From the above results, it can be presumed that form K is an anhydrous form.

As can be seen from fig. 11, X-ray powder diffraction peak data of form K is shown in table 27.

Watch 27

2θ(°)	Relative Strength (%)
		5.00	25.55
6.63	100.00
		8.18	12.52
10.01	9.31
		11.72	14.03
12.45	17.21
		13.28	15.71
15.15	10.38
		16.03	10.54
17.47	6.58
		18.22	18.49
19.29	25.54
		20.38	24.65
22.15	14.98
		22.83	7.05
24.79	5.61

3.12 form L

The form L sample (819805-14-C1 _ after 140 ℃) was obtained after heating the form C sample (819805-14-C1) to 140 ℃, and XRPD results for form L are shown in fig. 12. Repeated preparation of the crystal form L sample (819805-19-C _ AFT 140 ℃) is obtained by heating the crystal form C sample (819805-19-C) to 140 ℃, and the XRPD overlay is shown in figure 42. The TGA and DSC results for the sample of form L (819805-19-C _ AFT 140 ℃) are shown in FIG. 43, the TGA results indicate that there is a 0.3% weight loss of form L (819805-19-C _ AFT 140 ℃) when heated to 150 ℃; the DSC results showed that the sample had an endothermic peak at 189.1 ℃ (peak temperature) and an exothermic peak at 190.9 ℃ (peak temperature), presumably the sample crystallized as form B; the presence of an endothermic peak at 203.2 ℃ (starting temperature) is presumed to be caused by melting of form L. Taken together with the TGA/DSC data, form L is presumed to be the anhydrous form.

As can be seen from fig. 12, X-ray powder diffraction peak data of form L is shown in table 28.

Watch 28

In the screening and crystal identificationDuring the determination, 12 crystal forms are discovered in total, including initial crystal form A and crystal forms B, C, D, E, F, G, H, I, J, K and L, and X-ray powder diffraction (XRPD), thermogravimetric analysis (TGA), differential Scanning Calorimetry (DSC), hydrogen spectrum liquid nuclear magnetism(s) (TGA) ¹ H-NMR) and High Performance Liquid Chromatography (HPLC) to characterize a representative sample of the resulting crystalline form. The crystal form identification result shows that the crystal forms A, B, D, K and L are anhydrous crystal forms, the crystal form F is a hydrate, the crystal forms C and E are solvates, the repeated preparation of the crystal forms H, I and J is not successful (the crystal form B is obtained), and the crystal form G is transformed into the crystal form K in a dry state, so the crystal forms G, H, I and J are not identified.

4. Study of transformation relationship between crystal forms

The interconversion relationship studies were performed on each of the obtained forms, and the results are summarized in fig. 13. In order to determine the thermodynamic stability relationship between anhydrous crystal forms, suspension competition tests are carried out on related crystal forms. At room temperature (24 +/-3 ℃) and 50 ℃, the physical mixture of the anhydrous crystal forms A, B, D, K and L is respectively suspended and stirred in ACN and IPA for 2 days and then is converted into the anhydrous crystal form B, and the result shows that the thermodynamics of the crystal form B is more stable than that of the crystal forms A, D, K and L under the condition of room temperature to 50 ℃. Under the condition of room temperature, the physical mixture of the anhydrous crystal form B and the hydrate crystal form F is suspended and stirred for 2 days in different water activity conditions and then is converted into the anhydrous crystal form B, namely the hydrate crystal form F can be converted into the crystal form B under the condition of 0-1 water activity at room temperature. The specific study is as follows.

4.1 Anhydrous Crystal form transition relationship Studies

To investigate the stability relationship between the anhydrous crystalline forms a, B, D, K and L, suspension competition tests in ACN and IPA for each anhydrous crystalline form at room temperature and 50 ℃ were set up and the test results are summarized in table 29.

The method comprises the following specific steps: 1) Preparing a near-saturated solution of a starting sample (819805-01-A) in different solvents at a specified temperature; 2) Respectively adding samples (about 5mg each) of crystal forms A, B, D, K and L with equal mass into 2 ml of near-saturated solution to form suspension; 3) Suspension stirring at room temperature and 50 ℃ for about 2 days (800 rpm); 4) The solid was isolated and tested for XRPD. All experiments were finally transformed to form B anhydrous according to XRPD comparison in fig. 44, indicating that form B is thermodynamically more stable than forms a, D, K and L over the range of room temperature to 50 ℃.

TABLE 29 SUSPENSION COMPLEX TEST SUSPENSION OF ANHYDRATE FORMS

4.2 Anhydrous Crystal form and hydrate transformation relationship study

In order to research the transformation relation between the anhydrous crystal form B and the hydrate crystal form F, the crystal form B and the crystal form F are arranged in IPA/H at room temperature ₂ Suspension competition tests in O system at different water activities and the test results are summarized in Table 30.

The method comprises the following specific steps: 1) Preparing a near-saturated solution of a crystal form B sample (819805-14-B) in different water activity systems at room temperature; 2) Respectively adding equal mass of crystal form B and crystal form F samples (about 5mg each) into 1 ml of near-saturated solution to form suspension; 3) Suspension stirring at room temperature for about 2 days (-800 rpm); 4) The solid was isolated and tested for XRPD. According to the XRPD comparison in fig. 45 and 46, anhydrous form B was obtained under all water activity conditions, indicating that form F converted to form B at different water activities at room temperature.

TABLE 30 summary of test results for suspension competition of anhydrous crystalline forms and hydrates

5. Evaluation of form B

According to the test result of suspension competition, the crystal form B is more thermodynamically stable than the crystal forms A, D, K and L under the condition of room temperature to 50 ℃; hydrate form F is converted to form B at different water activities at room temperature; thus, form B was further evaluated as the dominant form, including solid state stability, hygroscopicity, and morphology.

5.1 solid State stability

To evaluate the solid state stability of form B, appropriate amounts of the samples were weighed and left closed at 80 ℃ for 1 day and open at 25 ℃/60% RH, 40 ℃/75% RH for one week, respectively. Solid samples under different conditions were tested for purity by HPLC to assess chemical stability and for crystal form by XRPD to assess physical stability. The results of the evaluations are summarized in table 31. The physical and chemical properties of form B remained stable under all three test conditions. The XRPD results are shown in FIG. 47.

Table 31 summary of solid state stability evaluation results for form B

* : relative purity = sample purity/initial state purity 100%.

Form B remains physically and chemically stable after 1 week of closed-mouth standing at 80 ℃ and 25 ℃/60% RH, 40 ℃/75% RH open-mouth standing for 1 week.

5.2 hygroscopicity index

The hygroscopicity of form B (819805-14-B) was evaluated by a dynamic moisture sorption test (DVS) at 25 ℃ and the results are shown in FIGS. 48 and 49. The results show that form B increased from 0% rh-80% rh by about 0.8%, indicating that form B is slightly hygroscopic and no transformation of the form occurs after DVS testing.

5.3 morphology

The sample of the crystal form B (819805-14-B) is subjected to a PLM test to evaluate the appearance of the sample, and the result is shown in figure 50, wherein the sample of the crystal form B is irregular particles and is partially agglomerated.

6. Pharmacokinetic studies of form a and form B

6.1 drugs and reagents

The crystal forms used in the research are a compound crystal form A and a compound crystal form B shown in a structural formula (I), and the sodium carboxymethyl cellulose is a medical supply grade.

6.2 test animals

The animals used in this study were 6 SD rats, male, weighing 150-200 g, divided into 2 groups of 3 animals on average.

6.3 preparation of the medicament

And weighing a proper amount of the crystal form A and the crystal form B, and adding sodium carboxymethylcellulose to ensure that the concentration of a test substance is 0.5%. The solid mixture was suspended in water to make a suspension with a final concentration of 10 mg/ml.

6.4 Experimental methods and results

Each suspension group was orally administered to fasted SD rats at a dose of 10ml/kg, and 100. Mu.l blood samples were collected via the tail vein at time intervals of 0.25, 0.5, 1, 2, 4, 6, 8, 24 hours after administration. Blood samples were collected in EDTA or heparin, centrifuged at 5000rpm for 5 minutes at 4 ℃ and plasma was collected.

After purification, the sample was analyzed by high performance liquid chromatography. The chromatographic condition is that 5mM NH4OAc-0.1% formic acid solution-methanol are used as a mobile phase, and the flow rate is 0.40mL/min. The PK profiles for form a and form B are shown in table 32 and figure 51.

Watch 32

It can be concluded that: the crystal form A and the crystal form B are easy to absorb, and have good blood concentration distribution in organisms and high bioavailability.

7. Conclusion of crystal form study

Taking the crystal form A of the compound shown in the structural formula (I) as an initial sample, and setting 100 polycrystal screening tests by adopting various methods. In the screening and crystal form identification processes, 12 different crystal forms (A-L) are discovered, including 5 anhydrous crystal forms A, B, D, K and L and a hydrate crystal form F. Suspension competition tests show that the anhydrous crystal form B is more thermodynamically stable than the crystal forms A, D, K and L under the condition of room temperature to 50 ℃; under the condition of room temperature water activity of 0-1, the hydrate crystal form F can be converted into an anhydrous crystal form B. The solid state stability test result shows that the crystal form B has better physical and chemical stability under the test condition. DVS testing indicated that form B was slightly hygroscopic. The PLM result shows that the crystal form B is irregular particles and is partially agglomerated.

And (3) combining the thermodynamic stability relationship and the solid state property evaluation result, recommending the crystal form B as a preferred crystal form for further development.

The crystal form B provided by the invention has stable physicochemical property and is very suitable for application in the aspects of research and development of medicines such as preparations in the future.

The crystal forms A, B, C, D, E, F, G, H, I, J, K and L provided by the invention can be used for preparing ALK inhibitor medicines, especially the crystal form B.

In the invention, the error range of the 2 theta angle value of the X-ray powder diffraction peak is +/-0.2 degrees. It should be understood that the 2 θ values of the X-ray powder diffraction pattern may vary slightly from machine to machine and from sample to sample, and that the numerical ranges may differ by within ± 0.2 units, and therefore the values quoted are not to be interpreted as absolute values.

In the present invention, the solvent name correspondence table is shown in the following table 33.

Watch 33

Claims

1. A crystalline form B of a compound of formula (I),

form B has an X-ray powder diffraction at 2 θ angle measured using Cu-ka radiation of: diffraction peaks are at 6.07 +/-0.2 degrees, 13.24 +/-0.2 degrees, 21.86 +/-0.2 degrees, 23.93 +/-0.2 degrees and 24.32 +/-0.2 degrees.

2. Form B of a compound of formula (i) according to claim 1, characterized in that form B has an X-ray powder diffraction at 2 Θ angles measured using Cu-ka radiation of: diffraction peaks are found at 9.38 +/-0.2 degrees, 17.37 +/-0.2 degrees, 18.34 +/-0.2 degrees, 19.64 +/-0.2 degrees, 20.10 +/-0.2 degrees, 22.23 +/-0.2 degrees, 23.39 +/-0.2 degrees, 26.61 +/-0.2 degrees, 27.65 +/-0.2 degrees, 29.82 +/-0.2 degrees, 30.52 +/-0.2 degrees, 31.64 +/-0.2 degrees, 33.65 +/-0.2 degrees, 35.72 +/-0.2 degrees and 36.81 +/-0.2 degrees.

3. Form B of the compound of formula (i) according to claim 1 or 2, having an X-ray powder diffraction pattern substantially as shown in figure 2, measured using Cu-K α radiation.

4. Crystalline form B of a compound of formula (i) according to claim 3, characterized in that its differential scanning calorimetry spectrum shows an endothermic peak at an onset temperature of 204.7 ℃.

5. Crystalline form B of a compound of formula (i) according to claim 3, characterized in that it is an anhydrous crystalline form.

6. A process for the preparation of form B of the compound of formula (i) as claimed in any one of claims 1 to 5, comprising the steps of:

s1: preparation of Crystal form A

Adding a compound shown in a formula (I) into water, adding dilute hydrochloric acid, stirring for dissolving, after complete dissolution, dropwise adding 0.2mol/L sodium hydroxide aqueous solution at 60 ℃ until the pH value of the system is 11-12, cooling to room temperature, filtering, and drying to obtain a crystal form A;

s2: preparation of form B

or, the crystal form A is subjected to gas-solid diffusion, gas-liquid diffusion, suspension stirring at 50 ℃, slow volatilization, anti-solvent addition, slow temperature reduction or high polymer induction to obtain a crystal form B;

the solvent used for gas-solid diffusion is selected from the following group: methanol, ethanol, acetone, tetrahydrofuran, 1, 4-dioxane and N, N-dimethylformamide;

the good solvent and the anti-solvent used for gas-liquid diffusion are selected from the following group:

good solvent Anti-solvent i Methylene dichloride Acetic acid isopropyl ester ii Ethanol Acetic acid isopropyl ester iii 1, 4-dioxane Acetic acid isopropyl ester iv Acetone (II) N-heptane v N, N-dimethyl acetamide Water (I) vi Methanol Methyl tert-butyl ether vii N-methyl pyrrolidone Methyl tert-butyl ether viii Trichloromethane Methyl tert-butyl ether

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The solvent used for 50 ℃ suspension stirring is selected from the following group: isopropyl acetate, isopropanol, methyl isobutyl ketone, ethyl acetate, methyl tert-butyl ether, acetonitrile, n-heptane, toluene, a solvent in a volume ratio of 1:4, a mixed solution of acetonitrile and water, wherein the volume ratio is 1:9, a mixed solution of methanol and methyl isobutyl ketone, and a solvent prepared from the following components in a volume ratio of 1:9, a mixed solution of acetone and isopropyl acetate, wherein the volume ratio of the acetone to the isopropyl acetate is 1:9, a mixed solution of dimethyl sulfoxide and isopropanol, wherein the volume ratio is 1:9 of a mixture of N, N-dimethylacetamide and methyl tert-butyl ether;

the solvent used for slow volatilization is selected from the group consisting of: acetone, isopropanol, ethyl acetate, 2-butanone, toluene, tetrahydrofuran;

the anti-solvent is added by using a good solvent and an anti-solvent selected from the following group:

good solvent Anti-solvent i Methanol Acetic acid isopropyl ester ii N-methyl pyrrolidone Acetic acid isopropyl esterEsters of salicylic acid iii Trichloromethane Acetic acid isopropyl ester iv 2-methyltetrahydrofuran Methyl tert-butyl ether v Acetone (II) Methyl tert-butyl ether vi Dimethyl sulfoxide N-heptane

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The solvent used for slow cooling is selected from the following group: 2-butanone, methyl isobutyl ketone, ethyl acetate, acetonitrile, toluene, and a solvent with a volume ratio of 1:1, a mixed solution of acetone and isopropyl acetate, wherein the volume ratio is 1:1, a mixed solution of tetrahydrofuran and n-heptane, wherein the volume ratio is 1:1, mixed solution of dimethyl sulfoxide and water;

the solvent and the high polymer used for inducing the high polymer are selected from the following group:

solvent(s) High polymer i Acetone (II) Mixed Polymer A ii Acetic acid ethyl ester Mixed Polymer A iii Methylene dichloride Mixed high polymer B

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The mixed high polymer A is an equal mass mixture of the following substances: polyvinyl pyrrolidone, polyvinyl alcohol, polyvinyl chloride, polyvinyl acetate, hydroxypropyl methylcellulose, and methylcellulose;

the mixed high polymer B is an equal mass mixture of the following substances: polycaprolactone, polyethylene glycol, polymethyl methacrylate, sodium alginate and hydroxyethyl cellulose.

7. Use of form B of a compound of formula (i) as claimed in any one of claims 1 to 5 in the manufacture of a medicament for inhibiting cell proliferation.

8. Use of form B of a compound of formula (i) as claimed in any one of claims 1 to 5 in the manufacture of a medicament for the treatment of cancer;

the cancer is selected from the group consisting of: lung cancer, anaplastic lymphoma, inflammatory myofibroblastoma, neuroblastoma.

9. Use of the compound of formula (i) as claimed in any one of claims 1 to 5 in its crystalline form B in the manufacture of a medicament for the inhibition of anaplastic lymphoma kinase.

10. Use of form B of a compound of formula (i) as claimed in any one of claims 1 to 5 in the manufacture of a medicament for the treatment of non-small cell lung cancer.

11. A pharmaceutical composition comprising a compound of formula (i) in crystalline form B according to any one of claims 1 to 5.