WO2022247772A1

WO2022247772A1 - Crystal forms of oxygen-containing heterocyclic compound, preparation method therefor and application thereof

Info

Publication number: WO2022247772A1
Application number: PCT/CN2022/094390
Authority: WO
Inventors: 许祖盛; 楼杨通; 唐佑海
Original assignee: 上海璎黎药业有限公司
Priority date: 2021-05-24
Filing date: 2022-05-23
Publication date: 2022-12-01
Also published as: CN115385923A

Abstract

Disclosed are crystal forms of an oxygen-containing heterocyclic compound, a preparation method therefor and an application thereof. The crystal forms include a crystal form A of a compound represented by formula (I), a crystal form B of the compound represented by formula (I), and a crystal form C of a solvate of the compound represented by formula (I). The crystal form preparation method is simple, and suitable for industrial production, and the crystal forms do not easily absorb moisture, have good stability, and are beneficial for the production of preparations and for the long-term storage of drugs.

Description

A crystal form of an oxygen-containing heterocyclic compound, its preparation method and application

This application claims the priority of Chinese patent application 2021105675408 with a filing date of May 24, 2021. This application cites the full text of the above-mentioned Chinese patent application.

technical field

The invention belongs to the field of medicine, and in particular relates to a crystal form of an oxygen-containing heterocyclic compound, a preparation method and an application thereof.

Background technique

Ras (Rat sarcoma viral oncogene, mouse sarcoma virus oncogene), was first discovered in rat sarcoma. Mammalian ras gene family has three members, which are H-ras, K-ras, and N-ras, and the fourth exon of K-ras has two variants, A and B. Ras genes are widely found in various eukaryotic organisms such as mammals, fruit flies, fungi, nematodes and yeasts, with varying degrees of expression in different tissues, among which H-Ras is mainly expressed in skin and skeletal muscle, and K-Ras is mainly expressed in skin and skeletal muscle. Expressed in the colon and thymus, N-Ras is highly expressed in the testis. As a molecular switch in the process of cell signal transduction, Ras protein regulates signal transduction by combining with GTP/GDP, and then regulates life processes such as cell proliferation, differentiation, aging and apoptosis.

Ras mutation is closely related to the occurrence and development of tumors. The Ras gene is mutated in more than 30 percent of human tumors and is considered one of the most potent drivers of cancer. Ras proto-oncogene mutations are mainly carried out through point mutations. More than 150 different Ras point mutations have been found, among which the mutations of glycine 12 and 13 and glutamine 61 are the most common.

For decades, people have been working on the development of small molecule inhibitors targeting Ras, but the development of related drugs has been slow. Scientists have always hoped to develop a GTP competitive inhibitor that directly acts on the Ras protein, but because of the strong affinity (pmol/L level) between GTP and Ras, the concentration of GTP in cells is very high (0.5mM), and Reasons such as the lack of pockets in the Ras protein structure that would facilitate small molecule binding were not successful. In recent years, people have made some progress in drug development using the allosteric site of the K-Ras G12C mutant. In 2013, a research team reported the discovery of small molecule inhibitors of K-Ras G12C (Nature, 2013, 503, 548-551). They identified a novel binding pocket located below the II region of the molecular switch from the K-Ras G12C mutant, and these inhibitors bound to this allosteric pocket and formed a covalent association with nearby Cys12 to selectively inhibit K-Ras Activation of G12C. Other researchers reported cell-active KRas inhibitors (Science, 2016, 351, 604-608). Amgen's compound Sotorasib (AMG 510) is connected to the 12th cysteine thiol of the KRAS-G12C mutant through the acrylamide Michael addition receptor structure, and locks the G12C mutant KRas protein in an inactive GDP binding state to specifically permanently and irreversibly inhibits its pro-proliferative activity. Sotorasib was approved by the FDA in May 2021, becoming the world's first targeted therapy for the treatment of patients with locally advanced or metastatic non-small cell lung cancer (NSCLC) carrying the KRas G12C mutation.

Those skilled in the art know that chemical drugs have polymorphism due to their different periodic arrangements of internal particles, and different solid forms of the same drug, including amorphous and crystalline forms, can exhibit very different physical, chemical and Differences in spectroscopic properties such as melting point, hygroscopicity, dissolution rate, solubility, processability, and stability lead to differences in safety and efficacy of drug products. Therefore, conducting an exhaustive study of drug polymorphism and evaluating the physicochemical properties of different solid forms of a drug to select the appropriate solid form for drug development has many practical benefits.

A kind of oxygen-containing heterocyclic compound, its structure is as shown in formula (I)

(hereinafter referred to as the compound represented by formula (I)), which can be used to treat and/or prevent various diseases mediated by Ras. The compound was independently designed and synthesized by the present inventors.

OJ column (20*250mm, 10μm; brand Daicel) was prepared by chiral resolution to obtain a solid form, which was determined to be amorphous by XRPD. Studies have found that when the compound represented by formula (I) exists in the form of an amorphous solid, it has the following defects: high moisture absorption; poor stability, especially under high temperature and light. The purity of the compound decreases significantly.

In view of the fact that the compound shown in formula (I) as found has poor potential to be developed as a drug when it exists in the form of an amorphous solid, the development of new crystal forms of the compound shown in formula (I) with more advantageous properties has very important practical significance.

Contents of the invention

The technical problem to be solved by the present invention is the defect that the solid form of the oxygen-containing heterocyclic compound is single, the hygroscopicity is high, and the stability is poor in the prior art. A crystal form of the oxygen-containing heterocyclic compound and its preparation method are provided. and applications. These crystal forms have simple preparation methods, are suitable for industrial production, are not easy to absorb moisture, have good stability, and are beneficial to the preparation of preparations and the long-term storage of medicines.

The present invention solves the above-mentioned technical problems through the following technical solutions.

One aspect of the present invention provides a crystal form A of the compound represented by formula (I), its X-ray powder diffraction pattern represented by 2θ angle, at 4.8±0.2°, 12.4±0.2°, 15.6±0.2° and 23.3 ± 0.2 ° have characteristic peaks, and the X-ray powder diffraction pattern is measured using the Kα spectral line of the Cu target;

In some embodiments, the crystal form A of the compound represented by formula (I), its X-ray powder diffraction pattern represented by 2θ angle, can be at 4.8±0.2°, 9.1±0.2°, 9.7±0.2 °, 12.4±0.2°, 15.6±0.2°, 19.8±0.2°, 20.4±0.2°, 21.1±0.2°, 23.3±0.2°, 24.7±0.2° and 26.3±0.2° have characteristic peaks.

In some embodiments, the crystal form A of the compound represented by formula (I) has an X-ray powder diffraction pattern represented by 2θ angles at 4.8±0.2°, 9.1±0.2°, 9.7±0.2° , 12.4±0.2°, 14.5±0.2°, 15.6±0.2°, 16.5±0.2°, 17.2±0.2°, 17.5±0.2°, 19.8±0.2°, 20.4±0.2°, 21.1±0.2°, 21.4±0.2° , 22.1±0.2°, 22.4±0.2°, 23.3±0.2°, 24.7±0.2°, 26.3±0.2°, 27.6±0.2° and 27.9±0.2° have characteristic peaks.

In some embodiments, the crystal form A of the compound represented by formula (I), its X-ray powder diffraction pattern represented by 2θ angle, its diffraction peak and peak area percentage can also be as follows:

Numbering	2θ(±0.2°)	Peak Area Percentage (%)
1	4.839	100.0
2	9.141	40.6
3	9.694	21.5
4	12.433	37.6
5	14.451	26.0
6	14.644	8.6
7	15.644	84.3
8	16.545	8.4
9	17.249	41.7
10	17.456	9.9
11	19.801	36.8
12	20.406	24.2
13	21.098	48.2
14	21.399	8.9
15	22.134	14.7
16	22.407	4.2
17	23.302	42.3
18	24.685	54.9
19	26.257	30.2
20	27.552	15.8
twenty one	27.948	17.6
twenty two	28.520	2.3
twenty three	29.742	2.2
twenty four	30.746	5.3

.

In some embodiments, the X-ray powder diffraction pattern represented by the 2θ angle of the crystal form A of the compound represented by formula (I) may also be substantially as shown in FIG. 3 .

In some embodiments, the differential scanning calorimetry (DSC) of the crystal form A of the compound represented by formula (I) may also have an endothermic peak at 167.7-171.0°C.

In some embodiments, the differential scanning calorimetry (DSC) of the crystal form A of the compound represented by formula (I) may also be substantially as shown in FIG. 4 .

In some embodiments, the thermogravimetric analysis (TGA) of the crystal form A of the compound represented by formula (I) may also be substantially as shown in FIG. 5 . In the TGA diagram, the Form A has substantially no weight loss before heating to the decomposition temperature, which shows that the Form A is an anhydrate.

In some embodiments, in the dynamic water sorption diagram (DVS) of the crystalline form A of the compound represented by formula (I), the increased weight of the crystalline form A compared to the initial weight can also be within 0 A weight gain of about 0.30% over the %-95% relative humidity range. It appears that the Form A is substantially non-hygroscopic.

In some embodiments, the dynamic moisture sorption diagram (DVS) of the crystal form A of the compound represented by formula (I) can also be substantially as shown in FIG. 6 .

In some embodiments, Form A of the compound represented by formula (I) is substantially pure. For example, the weight content of the crystal form A is at least 99%, at least 95%, or lower to 90%. Alternatively, the weight content of the crystal form A is at least 80%, or at least 70%, or lower to 60%. Or further, the weight content of the crystal form A reaches at least 50%.

Another aspect of the present invention provides a method for preparing the crystal form A of the compound represented by formula (I), which is method 1, method 2 or method 3:

Method 1: at 40°C to 80°C, cool the hot saturated solution formed by the compound represented by formula (I) and the solvent to room temperature, and crystallize; wherein, the solvent is selected from ethanol, ethyl acetate , one or more of methyl isobutyl ketone, acetonitrile and methyl tert-butyl ether;

Method 2: at 40°C to 80°C, mix the solution formed by the compound represented by formula (I) and solvent A with solvent B, and crystallize; wherein, the solvent A is ethanol, isopropanol, One or more of acetone, ethyl acetate, acetonitrile, tetrahydrofuran, 2-methyltetrahydrofuran and 1,4-dioxane, and the solvent B is selected from methyl tert-butyl ether, n-heptane and water one or more of

Method 3: Place an excess of the compound shown in formula (I) in a solvent to form a suspension. After the suspension is balanced, separate the solid-liquid phase and dry it; wherein, the solvent is selected from ethanol, methyl One or more of isobutyl ketone, methyl tert-butyl ether, acetonitrile, tetrahydrofuran, 2-methyltetrahydrofuran, and 1,4-dioxane.

In some embodiments, in the method 1, method 2 or method 3, the compound represented by formula (I) can be a compound represented by formula (I) in any solid form, for example, an amorphous Morphology A compound represented by formula (I) in the form of a solid.

in,

In method 1,

The amount of the solvent used can be the conventional amount used in this type of operation in the field, as long as a saturated solution is formed. In some embodiments, the volume-to-mass ratio of the solvent to the compound shown in formula (I) is 10.0 ~30.0 mL/g, eg, 16.7 mL/g.

The crystallization can be carried out by cooling at a rate of 5-30° C./hour.

In some embodiments, in the method 1, before the crystallization, the hot saturated solution is filtered. The filtration treatment method can be a conventional filtration method for such operations in the art, preferably thermal filtration.

The operation of solid-liquid separation and drying may also be included after the crystallization.

The solid-liquid separation can be filtering.

The drying may be drying under reduced pressure, for example, drying under reduced pressure at 40°C.

In method 2,

The volume-to-mass ratio of the solvent A to the compound represented by formula (I) is 8.0-11.0 mL/g, for example, 9.5 mL/g.

Said 40°C-80°C may be 70-75°C or 45-50°C.

The volume ratio of solvent A to solvent B is (0.2-5):1, for example, (0.38-1.9):1.

The timing of the mixing is not particularly limited, the solution formed by the compound shown in formula (I) and solvent A can be mixed with solvent B, or the solution formed by the compound shown in formula (I) and solvent A can be mixed Cool until a solid precipitates out and then mix with solvent B.

The mixing method can be dropwise, for example, add solvent B dropwise to the solution formed by the compound represented by formula (I) and solvent A.

In some embodiments, in the method 2, preferably, when solvent B is n-heptane, solvent A is ethanol and/or ethyl acetate; when solvent B is methyl tert-butyl ether, solvent A is selected from One or more of ethanol, isopropanol and 2-methyltetrahydrofuran.

In some embodiments, in the method 3, the solvent is selected from ethanol and/or methyl isobutyl ketone, for example, methyl isobutyl ketone.

In method 3, the amount of the solvent does not need to be limited, it can be conventional in the art, as long as it forms a suspension with the compound shown in formula (I), for example, when the solvent is methyl isobutyl When a base ketone is used, its volume-to-mass ratio to the compound shown in formula (I) is 0.2 g/mL.

The suspension equilibrium temperature can be conventional, as long as it is not higher than the boiling point of the solvent system, it can be 30°C to 60°C, for example, 50°C.

In some embodiments, in the method 3, the suspension equilibration time may depend on the scale of the reaction, generally within 2 hours to 14 days; for example, in a laboratory test, at room temperature, 2- It can be completed in about 24 hours; after amplification, the reaction time is extended, such as 24 hours to 14 days, preferably 7 days to 14 days, such as 7 days or 14 days.

One aspect of the present invention provides a crystal form B of the compound represented by formula (I), its X-ray powder diffraction pattern represented by 2θ angle, at 5.0±0.2°, 9.8±0.2°, 14.7±0.2° , 19.6±0.2°, 24.6±0.2° and 31.0±0.2° have characteristic peaks, and the X-ray powder diffraction pattern is measured using the Kα line of the Cu target;

In some embodiments, the crystal form B of the compound represented by formula (I), its X-ray powder diffraction pattern represented by 2θ angle, its diffraction peak and peak area percentage can also be as follows:

Numbering	2θ(±0.2°)	Peak Area Percentage (%)
1	4.958	100.0
2	9.838	19.8
3	14.718	1.1
4	19.631	4.6
5	24.582	2.9
6	31.047	1.1

.

In some embodiments, the X-ray powder diffraction pattern represented by the 2θ angle of the crystal form B of the compound represented by formula (I) may also be substantially as shown in FIG. 7 .

In some embodiments, the differential scanning calorimetry (DSC) of the crystal form B of the compound represented by formula (I) may also have an endothermic peak at 132.0-142.5°C.

In some embodiments, the differential scanning calorimetry (DSC) of the crystal form B of the compound represented by formula (I) may also be substantially as shown in FIG. 8 .

In some embodiments, the thermogravimetric analysis (TGA) of the crystal form B of the compound represented by formula (I) may also be substantially as shown in FIG. 9 . In the TGA diagram, the Form B has substantially no weight loss before heating to the decomposition temperature, which shows that the Form B is an anhydrate.

In some embodiments, in the dynamic water sorption diagram (DVS) of the crystalline form B of the compound represented by formula (I), the weight of the crystalline form B increased compared to the initial weight can also be within 0 A weight gain of about 0.95% over the %-95% relative humidity range. The Form B was shown to be substantially non-hygroscopic.

In some embodiments, the dynamic moisture sorption diagram (DVS) of the crystalline form A of the compound represented by formula (I) may also be substantially as shown in FIG. 10 .

In some embodiments, Form B of the compound represented by formula (I) is substantially pure. For example, the weight content of the crystal form B is at least 99%, at least 95%, or lower to 90%. Alternatively, the weight content of the crystal form B is at least 80%, or at least 70%, or lower to 60%. Or further, the weight content of the crystal form B reaches at least 50%.

Another aspect of the present invention provides a method for preparing crystal form B of the compound shown in formula (I), which is method A or method B:

Method A: at 50-70°C, form a hot saturated solution of the compound represented by formula (I) with methanol, and evaporate the solvent at room temperature for crystallization;

Method B: The compound represented by formula (I) is formed into a suspension with a solvent, and after the suspension is balanced, the solid-liquid phase is separated and dried; the solvent is methanol or "a mixture of methanol and water".

In some embodiments, in the method A or method B, the compound represented by formula (I) can be a compound represented by formula (I) in any solid form, for example, in the form of an amorphous solid The compound shown in formula (I).

In some embodiments, in the method A, before the crystallization of the volatile solvent, the hot saturated solution is filtered. The filtration treatment method can be a conventional filtration method for such operations in the art, preferably thermal filtration.

In some embodiments, in the method A, drying may also be included after the crystallization. Described drying is drying under reduced pressure.

In some embodiments, in the method B, when the solvent is a mixture of methanol and water, the volume ratio of methanol and water may be (5-95):5, such as 5:5 or 95:5.

The suspension equilibrium temperature can be conventional, as long as it is not higher than the boiling point of the solvent system, it can be 10°C to 40°C, for example, room temperature.

In some embodiments, in the method B, the suspension equilibration time depends on the scale of the reaction, generally within 2 days to 14 days; for example, in a laboratory test, at room temperature, 2 days It can be completed in about 10 minutes; after amplification, the reaction time is extended, such as 2 days to 14 days, preferably 7 days to 14 days, such as 7 days or 14 days.

In some embodiments, in the method B, the suspension can be heated while the heating temperature should not be higher than the boiling point of the solvent system, such as about 40°C, about 50°C. The heating can promote the transformation of the solid in the suspension into the crystal form B of the compound represented by formula (I).

Another aspect of the present invention also provides a crystal form C of the compound solvate shown in formula (I), the structure of the crystal form C is as follows:

It belongs to the triclinic crystal system, the space group is P1, and the unit cell parameters are:

α=95.12(3)°, β=93.82(3)°, γ=90.43(3)°; unit cell volume

The number of asymmetric units in the unit cell is Z=1.

In one embodiment, the crystal structure data of the crystal form C collected by X-ray single crystal diffraction are shown in Table 4 below:

Table 4

Another aspect of the present invention also provides a pharmaceutical composition, which comprises the above-mentioned crystal form A of the compound shown in formula (I), the above-mentioned crystal form B of the compound shown in formula (I) and the above-mentioned formula ( I) one or more of the crystal forms C of the solvates of the compounds shown, and pharmaceutically acceptable auxiliary materials.

In the present invention, the crystal form A of the compound shown in formula (I), the crystal form B of the compound shown in formula (I) and the crystal form C of the solvate of the compound shown in formula (I) One or more of them may also be used in combination with one or more other active ingredients; when used in combination, the active ingredients may be separate compositions for simultaneous administration in therapy by the same or different routes of administration or They are administered separately at different times, or they can also be administered together in the same pharmaceutical composition.

In the present invention, the administration method of the pharmaceutical composition is not particularly limited, and preparations in various dosage forms can be selected for administration according to the patient's age, gender, and other conditions and symptoms; for example, tablets, pills, solutions, suspensions, Emulsions, granules or capsules are administered orally; injections can be administered alone, or mixed with delivery fluids for injection (such as glucose solution and amino acid solution) for intravenous injection; suppositories are administered to the rectum.

Another aspect of the present invention also provides the use of the compound represented by formula (I) or the above pharmaceutical composition in the preparation of medicines for treating and/or preventing diseases mediated by Ras.

In some embodiments, the compound represented by formula (I) is crystal form A of the compound represented by formula (I), crystal form B of the compound represented by formula (I) above, or the compound represented by formula (I) above. Form C of the solvate of the compound represented by formula (I).

In some embodiments, the Ras is, for example, a G12C mutation of one or more of K-Ras, H-Ras and N-Ras, and another example is a G12C mutation of K-Ras.

In some embodiments, the Ras-mediated disease is, for example, cancer. Such cancers as colon cancer, appendix cancer, pancreatic cancer, MYH-associated polyposis, blood cancer, breast cancer, endometrial cancer, gallbladder cancer, bile duct cancer, prostate cancer, lung cancer, brain cancer, ovarian cancer, cervical cancer Cancer, testicular cancer, kidney cancer, head or neck cancer, bone cancer, skin cancer, rectal cancer, liver cancer, esophagus cancer, stomach cancer, thyroid cancer, bladder cancer, lymphoma, leukemia and melanoma.

Another aspect of the present invention also provides the application of the above-mentioned compound represented by formula (I) or the above-mentioned pharmaceutical composition in the preparation of Ras inhibitors.

In some embodiments, the Ras inhibitor can be used in mammalian organisms; it can also be used in vitro, mainly for experimental purposes, for example: as a standard sample or control sample to provide comparison, or prepared according to conventional methods in the art A kit that provides rapid detection of Ras inhibitory effects.

Another aspect of the present invention also provides the application of the above-mentioned compound represented by formula (I) or the above-mentioned pharmaceutical composition in the preparation of medicaments for treating and/or preventing cancer.

In some embodiments, the cancer is, for example, colon cancer, appendix cancer, pancreatic cancer, MYH-associated polyposis, blood cancer, breast cancer, endometrial cancer, gallbladder cancer, bile duct cancer, prostate cancer, lung cancer, brain cancer , ovarian cancer, cervical cancer, testicular cancer, kidney cancer, head or neck cancer, bone cancer, skin cancer, rectal cancer, liver cancer, esophagus cancer, stomach cancer, thyroid cancer, bladder cancer, lymphoma, leukemia and melanoma one or more.

Unless otherwise specified, terms used in the present invention have the following meanings:

The "compound shown in formula (I)" used in the present invention refers to the compound with the chemical structure shown in the following formula (I):

"Amorphous" or "amorphous form" as used in the present invention means that the solid form of the compound represented by formula (I) is a non-crystalline form.

"Crystal form", "crystal form" and "polymorph" used in the present invention can be used interchangeably in the present invention, specifically referring to the crystal form A of the compound shown in formula (I), the above-mentioned Crystal form B of the compound represented by formula (I) or crystal form C of the solvate of the compound represented by formula (I).

The crystal form of the present invention can be identified by one or several solid-state analysis methods. Such as X-ray powder diffraction, single crystal X-ray diffraction, infrared absorption spectrum, differential scanning calorimetry, thermogravimetric curve, etc. Those skilled in the art know that the peak intensity and/or peak situation of X-ray powder diffraction may vary due to different experimental conditions. At the same time, due to the different accuracy of the instrument, the measured 2θ value will have an error of about ±0.2°. The relative intensity value of the peak is more dependent on certain properties of the measured sample than the position of the peak, such as the size of the crystal and the degree of purity, so the measured peak intensity may have a deviation of about ± 20%. Although there are experimental errors, instrumental errors, orientation priorities, etc., those skilled in the art can still obtain sufficient information to identify each crystal form from the X-ray powder diffraction data provided by this patent. In the measurement of infrared spectroscopy, the shape of the spectrum and the position of the absorption peak will be affected to a certain extent due to the different performance of various types of instruments, the difference in the degree of grinding or the degree of water absorption of the test product during preparation, etc. However, in DSC measurement, the initial temperature, maximum temperature and heat of fusion data of the endothermic peak obtained by actual measurement have a certain degree of variability according to the heating rate, crystal shape and purity and other measurement parameters.

"Anhydrous" used in the present invention means that the sample contains no more than 1.0% (weight percent) or no more than 0.5% (weight percent) of water as determined by TGA.

"About" used in the present invention is used for a certain parameter, such as quantity, angle, temperature, time, etc., and means to deviate from the specified value by at most ±10%, preferably within ±5%, and most preferably within ±2%; however, when measuring the onset temperature and peak temperature of a thermal event in a differential scanning calorimetry (DSC) diagram, the term "about" means The onset or peak temperature can typically be within ±3°C. As will be understood by those skilled in the art, the use of a number for a non-critical parameter is for illustrative purposes only, not limitation.

As used herein, "substantially pure" when used to describe a polymorphic form of a compound as represented by formula (I), means that the solid form of the compound contains this polymorphic form and substantially does not contain Other polymorphs of said compounds. Typical substantially pure polymorphs contain less than 50%, preferably less than 40%, preferably less than 30%, preferably less than 20%, preferably less than 10%, preferably less than 5% by weight of other polymorphs. %, preferably less than 1%.

When the "substantially non-hygroscopic" used in the present invention is used to describe the polymorphic form of the compound shown in formula (I), it means that when using the dynamic moisture adsorption (DVS) technique to determine the polymorphism shown in formula (I), The polymorphic form of the compound has an increased mass of less than 2%, preferably less than 1%, compared to the initial mass in the range of 0-95% relative humidity.

In the present invention, the "heated saturated liquid" refers to heating a solute in one or several solvents to form a supersaturated solution, wherein the solute is in excess. The heating temperature is usually about 5-30°C below the boiling point of the solvent, for example, the heating temperature can be 40°C, 50°C, 60°C, 70°C, 75°C, 80°C.

In the present invention, the "hot filtration" used refers to filtering the hot saturated liquid through a pinhole filter, wherein the filter membrane material is preferably polyvinylidene fluoride (PVDF) or nylon, and the filter membrane pore size is preferably is 0.45 microns.

In the present invention, the "hot clarified solution" used refers to the complete dissolution of the solute in one or several solvents by heating, wherein the solvent is in excess. The heating temperature is usually about 5-30°C below the boiling point of the solvent, for example, the heating temperature can be 40°C, 50°C, 60°C, 70°C, 75°C, 80°C.

In the present invention, the "suspension" used refers to forming a supersaturated solution of a solute in one or several solvents, wherein the solute is in significant excess, such as the suspension prepared in a suspension equilibrium experiment.

In the present invention, the "suspension equilibrium" used can be completed by conventional methods in the art, for example, by rotating the prepared suspension vial 360°, or by stirring the prepared suspension way. The temperature of said suspension equilibrium usually refers to room temperature unless otherwise specified.

In the present invention, the "room temperature" refers to 10-30°C.

In the present invention, the operation of "solid-liquid phase separation" can be accomplished by conventional methods in the art, such as filtration and centrifugation. When the "solid-liquid phase separation" is completed by a filtration operation, the filtration, unless otherwise specified, refers to suction filtration under reduced pressure. The specific operation of the centrifugation is: place the pre-separated sample in a centrifuge, the centrifugation speed is usually 3000-15000 rpm, preferably 6000-12000 rpm. The solid obtained through the "separation" can be further washed, and the solvent used for washing is preferably the same as that used in the crystal preparation method, and the amount of the washing solvent is generally 0.1-1 times the volume of the solvent used in the crystal preparation method.

In the present invention, the "drying" can be accomplished by conventional methods in the art, such as drying under normal pressure, drying under reduced pressure, preferably drying under reduced pressure. Without limitation, the vacuum degree of the vacuum drying may be -0.09MPa; the temperature of the vacuum drying may be 30-80°C, preferably 40-70°C, such as 40°C, 50°C. The drying time is generally 1 hour to overnight, that is, 1-24 hours, such as 2 hours, 16 hours.

In the present invention, the "stirring" can be accomplished by conventional methods in the field, such as magnetic stirring, mechanical stirring, and the stirring speed is 50-1200 rpm, preferably 200-500 rpm.

In the present invention, the used "volatile" refers to covering the mouth of the vial filled with the solution filtered by the filter membrane with aluminum foil and piercing a small hole, and then slowly volatilizing the solvent in a laboratory environment, for example, the bottle containing The mouth of the vial with the solution filtered by the filter membrane is covered with aluminum foil and a small hole is pierced, and the solvent is slowly evaporated in the fume hood.

In the present invention, "treating" refers to ameliorating a disease or disorder (i.e. arresting the disease or reducing the manifestations, extent or severity of its clinical symptoms); alternatively, improving at least one physical parameter, which may not be perceived by the subject; or slowing down Disease progression.

In the present invention, "prevention" means a reduction in the risk of acquiring or developing a disease or disorder (i.e. resulting in the absence of at least one of the clinical symptoms of the disease in subjects who may have been exposed to an agent causing the disease or susceptible to the disease before onset of the disease) .

The term "pharmaceutically acceptable excipients" refers to the excipients and additives used in the production of medicines and formulation of prescriptions, and refers to all substances contained in pharmaceutical preparations except active ingredients. See Pharmacopoeia of the People's Republic of China (2020 edition) four, or, Handbook of Pharmaceutical Excipients (Raymond C Rowe, 2009 Sixth Edition).

On the basis of not violating common knowledge in the field, the above-mentioned preferred conditions can be combined arbitrarily to obtain preferred examples of the present invention.

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

The positive progress effect of the present invention is:

The crystal form of the compound represented by formula (I) provided by the present invention has one or more improved properties, especially in the properties of good purity, improved stability to high temperature and light, and improved high humidity conditions Hygroscopicity, not easy to crystallization, can better resist the problems of the purity of active ingredients and the growth of impurities caused by environmental temperature, humidity, light and other factors in the process of drug production, storage, transportation, etc., and reduce the resulting The risk of curative effect decline and safety risk; and the ability to better resist the risk of changes in the bioavailability of the drug due to the transformation of the solid form of the active ingredient during long-term storage of the drug preparation, and is suitable as an active ingredient for drug formulation development. The preparation method provided by the invention is simple and easy to operate, and is suitable for industrial production.

Description of drawings

Fig. 1 is an X-ray powder diffraction (XRPD) pattern of an amorphous form of the compound represented by formula (I).

Fig. 2 is the unimolecular three-dimensional structure ellipsoid diagram of the crystal form C of the compound solvate shown in formula (I).

Fig. 3 is the XRPD spectrum of the crystal form A of the compound represented by formula (I).

Fig. 4 is a differential scanning calorimetry (DSC) diagram of the crystal form A of the compound represented by formula (I).

Fig. 5 is a thermogravimetric analysis (TGA) diagram of the crystal form A of the compound represented by formula (I).

Fig. 6 is a dynamic moisture adsorption (DVS) diagram of the crystal form A of the compound represented by formula (I); wherein 1 is the hygroscopic curve, and 2 is the dehydration curve.

Fig. 7 is the XRPD spectrum of the crystal form B of the compound represented by formula (I).

Fig. 8 is a DSC chart of the crystal form B of the compound represented by formula (I).

Fig. 9 is a TGA diagram of the crystal form B of the compound represented by formula (I).

Figure 10 is the DVS diagram of the crystal form B of the compound represented by formula (I); wherein 1 is the hygroscopic curve, and 2 is the dehydration curve.

Figure 11 is a DVS diagram of the amorphous form of the compound represented by formula (I); wherein 1 is the hygroscopic curve, and 2 is the dehydration curve.

Fig. 12 is a graph showing the body weight changes of mice in the subcutaneous xenograft tumor model of lung cancer cell NCI-H358.

Fig. 13 is a diagram showing the change of tumor volume in mice in the subcutaneous xenograft tumor model of lung cancer cell NCI-H358.

Fig. 14 is a graph showing the body weight changes of mice in the pancreatic cancer cell MIA PaCa-2 subcutaneous xenograft tumor model.

Fig. 15 is a graph showing the changes in tumor volume in mice in the pancreatic cancer cell MIA PaCa-2 subcutaneous xenograft tumor model.

Fig. 16 is a graph showing changes in body weight of mice in the subcutaneous xenograft tumor model of human-derived colon cancer cell SW837.

Fig. 17 is a diagram showing the change of mouse tumor volume in the subcutaneous xenograft tumor model of human colon cancer cell SW837.

Detailed ways

The present invention is further illustrated below by means of examples, but the present invention is not limited to the scope of the examples. For the experimental methods that do not specify specific conditions in the following examples, select according to conventional methods and conditions, or according to the product instructions.

The following examples employ the compound represented by formula (I) in the form of an amorphous solid as a starting material.

The solvents involved in the following examples are analytically pure or chromatographically pure, and when the solvents are mixed solvents, unless otherwise specified, all are volume ratios.

Used test instrument and test condition of experiment among the present invention:

(1) X-ray powder diffraction (X-Ray Powder Diffraction, XRPD)

Bruker's D8Advance X-ray powder diffractometer is used for detection, and the Kα line of the Cu target is used

The voltage is 40 kV, the current is 40 mA, the divergence slit is 1.0mm, the solar slit is 0.4°, the scanning mode is continuous scanning, the scanning angle range is 3°-45°, the step size is 0.02°, and the scanning speed is 8 °/min, detector: LynxEye.

(2) Differential Scanning Calorimeter (DSC)

The DSC25 differential scanning calorimeter of TA Instruments was used for detection, the atmosphere was nitrogen, the heating rate was 10°C/min, and the temperature rising range was 25-300°C.

(3) Thermogravimetric Analysis (Thermo Gravimetric Analysis, TGA)

The Q500 thermogravimetric analyzer of TA Instruments was used for detection, the atmosphere was nitrogen, and the temperature was heated to 350° C. at a heating rate of 10° C./min.

(4) Dynamic Vapor Sorption (DVS)

The Advantage 1.0 dynamic moisture adsorption instrument of SMS company is used for detection, the temperature is 25°C, the relative humidity range is 0%-95%, and the humidity change step is 5% relative humidity. When the value of the mass change rate dm/dt is less than 0.002% When the balance is considered to be balanced, when the mass change rate is less than 0.01%/min within 5 minutes, it is the balance standard in the detection process, and the longest balance time is 2 hours.

Preparation Example 1 Synthesis of the compound shown in formula (I)

(1) Synthesis of Compound 1

The synthetic route of compound 1:

Synthesis of compound 1-j

The compound 1-bromo-8-chloronaphthalene (500mg, 2.07mmol) was dissolved in THF (20mL), cooled to -78°C, and n-BuLi (2.5M, 1.66mL, 4.14mmol) was added dropwise under nitrogen protection . After the dropwise addition was completed, the mixture was stirred at -78°C for 10 minutes, and then DMF (800 μL, 10.35 mmol) was added dropwise at -78°C. After the addition was complete, the reaction mixture was stirred at -78°C for 30 minutes, then raised to room temperature and stirred for 2 hours, quenched with 50 mL of saturated ammonium chloride solution, and extracted with ethyl acetate (50 mL*2). The organic phase was washed with saturated brine (50 mL*2), treated with anhydrous sodium sulfate, filtered, and concentrated to obtain a crude product. The crude product was separated and purified by flash separation column (EA/PE=1/10) to obtain compound 1-j (330 mg, 84% yield) as a white solid. LC-MS (ESI): m/z = 191.0[M+H] ⁺ ; ¹ H NMR (400MHz, CDCL ₃ ): δ11.31 (s, 1H), 8.03 (dd, 1H, J ₁ = 1.2Hz, J ₂ =8.4Hz), 7.92(dd,1H, J ₁ =1.2Hz, J ₂ =7.2Hz), 7.86(1H,J=8.4Hz),7.70(dd,1H,J ₁ =1.2Hz,J ₂ =7.6Hz),7.59(t,1H,J=7.6Hz),7.47(t,1H,J=8Hz).

Synthesis of compound 1-i

NaH (60%, 242 mg, 6.05 mmol) was added to 6 mL of THF at room temperature. Then methyl acetoacetate (543μL, 5.04mmol) was added under nitrogen at room temperature, and the mixture was stirred at room temperature under nitrogen for 30 minutes, and n-BuLi (2.5M, 2.4mL , 6.05mmol). After the addition was complete, the mixture was kept at this temperature for 30 minutes, and then a solution of compound 1-j (320 mg, 1.68 mmol) in THF (10 mL) was added dropwise. After the addition was complete, the mixture was stirred at low temperature (-10°C-0°C) for 2 hours, quenched with saturated ammonium chloride solution (50 mL), and then extracted with ethyl acetate (50 mL*2). The organic phase was washed with saturated brine (50mL*2), treated with anhydrous sodium sulfate, filtered, and concentrated to obtain a crude product, which was separated and purified by a flash separation column (EA/DCM=1/10) to obtain compound 1-i (510 mg, 99% yield) as a white solid. LC-MS (ESI): m/z=329.1[M+Na] ⁺ ; ¹ HNMR (400MHz, CDCl ₃ ): δ8.06(d, 1H, J=6.4Hz), 7.79(d, 2H, J= 8Hz), 7.58(dd, 1H, J ₁ = 7.6Hz, J ₂ = 1.6Hz), 7.53(t, 1H, J = 7.6Hz), 7.34(t, 1H, J = 7.6Hz), 6.91(dd, 1H, _J1 = 9.2Hz, _J2 = 2.4Hz), 3.74(s, 3H), 3.54(s, 2H), 3.36(dd, 1H, _J1 = 18Hz, _J2 = 1.6Hz), 3.24(d ,1H,J＝3.6Hz),2.85-2.75(m,1H).

Synthesis of compound 1-h

Compound 1-i (510 mg, 1.66 mmol) was dissolved in DCM (18 mL) at room temperature, and then DMF-DMA (245 μL, 1.83 mmol) was added at room temperature under nitrogen. After the reaction solution was stirred for 45 minutes at room temperature, BF ₃ ·Et ₂ O (232 μL, 1.83 mmol) was added. After the addition was complete, the mixture was stirred at room temperature for 1 hour and then diluted with 100 mL of ethyl acetate. The organic phase was washed successively with saturated NaHCO ₃ solution (100 mL) and saturated brine (100 mL*2), treated with anhydrous sodium sulfate, filtered and concentrated to obtain crude compound 1-h (520 mg). The crude product was directly used in the next reaction without further purification. LC-MS(ESI): m/z=317.1[M+1] ⁺ .

Synthesis of compound 1-g

Compound 1-h (520mg, 1.64mmol) was dissolved in THF (20mL) at room temperature, and tri-sec-butyllithium borohydride (1M, 1.64mL, 1.64mmol) was added dropwise at -78°C under nitrogen ). After the addition was complete, the mixture was stirred at -78°C for 1 hour, quenched with saturated ammonium chloride solution (50 mL), extracted with ethyl acetate (50 mL*2), and washed with saturated brine (50 mL*2). Treatment with anhydrous sodium sulfate, filtration, and concentration gave the crude product, which was separated and purified by a flash separation column (PE/EA=4/1) to obtain compound 1-g (338 mg, 65% yield) as a yellow oil. LC-MS(ESI): m/z=319.0[M+1] ⁺ .

Synthesis of compound 1-f

At room temperature, compound 1-g (338mg, 1.06mmol) was dissolved in methanol (20mL), and then sodium methoxide (286mg, 5.3mmol) and compound 2-methyl-2 were added successively under nitrogen at 0°C. - Urea mercaptosulfate (265 mg, 0.954 mmol). After the addition was complete, the mixture was warmed to room temperature and stirred for 20 hours. The pH of the reaction solution was adjusted to 5 with 1N dilute hydrochloric acid, the solid precipitated out, filtered, the filter cake was washed with water (5mL*2), the solid was collected, and dried in vacuo to obtain the crude product 1-f (313mg) as a white solid. LC-MS(ESI): m/z=359.1[M+1] ⁺ .

Synthesis of compound 1-e

Compound 1-f (313 mg, 0.87 mmol) was dissolved in DCM (10 mL) at room temperature, and then DIPEA (431 μL, 2.61 mmol), trifluoromethanesulfonic anhydride (219 μL, 1.31 mmol). After the addition was complete, the reaction mixture was stirred for 2 hours in an ice-water bath, quenched with saturated sodium bicarbonate solution (50 mL), extracted with DCM (50 mL*2), and the organic phase was treated with anhydrous sodium sulfate, filtered, and concentrated to obtain The crude product was separated and purified by flash separation column (EA/PE=1/10) to obtain compound 1-e (83 mg, 16% yield in 2 steps) as a white solid. LC-MS(ESI): m/z=491.0[M+1] ⁺ .

Synthesis of compound 1-d

Dissolve compound 1-e (83mg, 0.169mmol) in DMF (10mL) at room temperature, then add DIPEA (84μL, 0.507mmol), (S)-2-cyanomethylpiperazine-1-carboxylic acid Benzyl ester hydrochloride (59.9 mg, 0.203 mmol). After the addition was complete, the mixture was stirred at 100 °C under nitrogen protection for 1 hour, then cooled to room temperature, quenched with saturated brine (50 mL), and extracted with ethyl acetate (50 mL*2). The organic phase was washed with saturated brine (50mL*3), then treated with anhydrous sodium sulfate, filtered, and concentrated to obtain a crude product, which was separated and purified by a flash separation column (EA/PE=1/1) to obtain compound 1-d (101 mg, 99% yield) as a white solid. LC-MS(ESI): m/z=600.2[M+1] ⁺ .

Synthesis of compound 1-c

Compound 1-d (101 mg, 0.168 mmol) was dissolved in ethyl acetate (10 mL) at room temperature, and then MCPBA (85%, 88.4 mg, 0.437 mmol) was added at room temperature. After the addition was completed, the mixture was stirred at room temperature for 2 hours, quenched with saturated sodium bicarbonate solution (20 mL), extracted with ethyl acetate (25 mL*2), and the organic phase was treated with anhydrous sodium sulfate, filtered, and concentrated to obtain a crude product. The crude product was separated and purified by flash separation column (EA/PE=1/4) to obtain compound 1-c (88 mg, 82% yield) as a white solid. LC-MS(ESI): m/z=632.1[M+1] ⁺ .

Synthesis of compound 1-b

At room temperature, compound 1-c (88mg, 0.139mmol) was dissolved in toluene (10mL), then the reaction solution was cooled to 0°C, N-methylprolinol (29μL, 0.243mmol) was added successively, t- BuONa (27 mg, 0.278 mmol). After the addition was complete, the reaction mixture was stirred in an ice-water bath under nitrogen for 0.5 hours, quenched with water (20 mL), and extracted with ethyl acetate (30 mL*2). The organic phase was treated with anhydrous sodium sulfate, filtered, and concentrated to obtain a crude product, which was separated and purified by a flash column (MeOH/DCM=1/10) to obtain compound 1-b (78 mg, 84% yield) as a white solid. LC-MS(ESI): m/z=667.3[M+1] ⁺ .

Synthesis of compound 1-a

Dissolve compound 1-b (72mg, 0.108mmol) in methanol (50mL) at room temperature, then cool the reaction solution to -78°C, replace with nitrogen twice, then add Pd/C (150mg), ZnBr ₂ (24.3mg, 0.108mmol), replaced with hydrogen 3 times, the reaction solution was raised to room temperature, and stirred for 5 hours under hydrogen. The reaction solution was filtered and concentrated to obtain a crude product, which was separated and purified by a flash column (MeOH/DCM=1:4) to obtain compound 1-a (20 mg, 35% yield) as a white solid. LC-MS(ESI): m/z=533.0[M+1] ⁺ .

Synthesis of compound 1

Dissolve the compound 2-fluoroacrylic acid (5.1 mg, 0.0563 mmol) in DMF (2 mL) at room temperature, then add HATU (25.6 mg, 0.0675 mmol), DIPEA (18.6 μL, 0.113 mmol), after the addition was complete, the reaction mixture was stirred at 0°C under nitrogen for 20 minutes, and a DMF (3 mL) solution of compound 1-a (20 mg, 0.0375 mmol) was added to the above reaction solution, warmed to room temperature, and continued to stir for 5 hours. Quenched with saturated brine (20mL), extracted with ethyl acetate (25mL*2), washed the organic phase with saturated brine (50mL*3), treated with anhydrous sodium sulfate, filtered and concentrated to obtain the crude product, which was passed through PREP- TLC separation and purification (MeOH/DCM=1/10) gave compound 1 (6 mg, 26% yield) as a white solid. LC-MS(ESI):m/z=605.2[M+1] ⁺ ; ¹ H NMR(400MHz, CDCl ₃ ):δ7.99-7.93(m,1H),7.83(t,2H,J=8.8Hz ),7.62-7.49(m,2H),7.36(t,1H,J=7.6Hz),6.55-6.44(m,1H),5.51-5.31(m,1H),5.25(d,1H,J=16.8 Hz), 5.02-4.93(m,1H), 4.82(dd,1H, J ₁ =2.4Hz, J ₂ =13.6Hz), 4.48-4.38(m,1H), 4.32-4.19(m,1H), 4.17 -4.04(m,1H),4.00(d,1H,J＝14Hz),3.87-3.70(m,1H),3.66-3.36(m,2H),3.31-3.16(m,2H),3.14-2.98( m,1H),2.96-2.69(m,4H),2.59(d,3H,J＝18Hz),2.52-2.34(m,1H),2.15-2.06(m,1H),1.87-1.74(m,2H ),0.93-0.76(m,2H).

(2) Resolution of Compound 1

Synthesis of compounds 1-1 and 1-2

The chiral resolution of compound 1 to obtain the compound shown in formula (I) is the difficulty. Despite trying a variety of conditions, the two isomers of compound 1 were inseparable on the TLC plate and could not be separated by TLC means; even in HPLC, the two isomers of compound 1 were separated Degree is also very poor, can't realize separation by preparative HPLC; Have to resort to chiral resolution at last, tried a plurality of conditions (as following table 1), found chiral resolution condition 9 at last, realized pair as formula (I ) Separation of the compound shown in ) and its diastereomers.

Table 1

The newly prepared compound 1 (260mg, 0.43mmol) was subjected to chiral resolution under the conditions shown in Table 2 below. Compound 1-1 (76 mg, 29% yield) was obtained as a white solid; Compound 1-2 (67 mg, 26% yield) was obtained as a white solid. Among them, according to the single crystal structure analysis of the compound represented by the formula (I) as shown below, it can be determined that the compound 1-2 is the compound represented by the formula (I).

Table 2

1-1: LC-MS (ESI): m/z=605.3[M+1] ⁺ ; ¹ H NMR (400MHz, CDCl ₃ ) δ7.96(d, 1H, J=7.2Hz), 7.83(t, 2H, J = 8.4Hz), 7.65-7.50 (m, 2H), 7.36 (t, 1H, J = 8.0Hz), 6.47 (dd, 1H, J ₁ = 10.8Hz, J ₂ = 3.2Hz), 5.42 ( d,1H,J=49.2Hz),5.26(dd,1H,J ₁ =3.6Hz,J ₂ =16.8Hz),5.05-4.76(m,1H),4.97(d,1H,J=13.6Hz), 4.84 (d, 1H, J = 13.6Hz), 4.36 (dd, 1H, J ₁ = 4.8Hz, J ₂ = 10.4Hz), 4.17 (dd, 1H, J ₁ = 6.8Hz, J ₂ = 10.8Hz), 4.06-3.87 (m, 1H), 3.77 (d, 1H, J = 10Hz), 3.59 (dd, 1H, J ₁ = 2.4Hz, J ₂ = 17.6Hz), 3.50-3.15 (m, 3H), 3.14- 2.99(m,2H),2.96-2.82(m,2H),2.72-2.59(m,1H),2.47(s,3H),2.32-2.21(m,1H),2.10-1.98(m,1H), 1.89-1.67(m,4H).

1-2: LC-MS (ESI): m/z=605.2[M+1] ⁺ ; ¹ H NMR (400MHz, CDCl ₃ ) δ7.97(d, 1H, J=7.2Hz), 7.83(t, 2H, J = 9.2Hz), 7.63-7.51 (m, 2H), 7.36 (t, 1H, J = 7.6Hz), 6.52 (dd, 1H, J ₁ = 3.2Hz, J ₂ = 10.8Hz), 5.42 ( d,1H,J=47.2Hz),5.25(dd,1H, _J1 =3.6Hz, _J2 =16.4Hz),4.99(d,1H,J=14.0Hz),4.82(d,1H,J=13.6 Hz), 5.05-4.72(m,1H), 4.38(dd,1H, J ₁ =4.8Hz, J ₂ =10.4Hz), 4.15(dd,1H, J ₁ =6.8Hz, J ₂ =10.8Hz), 3.98(d,1H,J=14Hz),3.87-3.73(m,1H),3.60(dd,1H, _J1 =2.4Hz, _J2 =18.4Hz),3.66-3.54(m,1H),3.54- 3.41(m,1H),3.16-2.98(m,2H),2.95-2.71(m,3H),2.71-2.61(m,1H),2.46(s,3H),2.33-2.19(m,1H), 2.10-1.98(m,1H),1.90-1.66(m,4H).

It was confirmed by XRPD that the solid form of compound 1-2 (compound represented by formula (I)) prepared by the above method was amorphous, and its XRPD spectrum was shown in FIG. 1 .

Example 1 Preparation of Crystal Form C of Compound Solvate Shown in Formula (I)

Another difficulty in this work is the identification of the structure of the compound shown in formula (I). The compounds represented by formula (I) and their diastereoisomers have obvious differences in in vitro cell activity, and it is difficult to confirm the chiral structure by chemical, general spectroscopy and other methods. Finally, X-ray diffraction technology was turned to for help. For this reason, several conditions were tried to cultivate single crystals, but all failed. Finally, fortunately, through the solvent evaporation method, under the solvent methanol-water (20:1) condition (the condition corresponding to No. 7 in Table 3 below), a colorless and transparent crystal that meets the requirements of the single crystal X-ray diffraction experiment was obtained. Flake crystals. Through analysis, it is determined that compound 1-2 is the compound shown in formula (I), and compound 1-1 is its diastereoisomer.

Single crystal cultivation process: the single crystal cultivation experiment was carried out at room temperature using the methods and solvent conditions shown in Table 3 below.

table 3

Single crystal test conditions: Measured according to the first method of "Chinese Pharmacopoeia" 2020 Edition Four General Rules 0451, using Bruker's D8 Venture single crystal X-ray diffractometer, MoKα radiation, ω scan. The total number of diffraction points collected in the single crystal diffraction experiment is 18322, the number of independent diffraction points is 10763, and the number of observable points (|F| ² ≥ 2σ|F| ² ) is 7231.

Analysis of single crystal structure: using direct method (Shelxs97) to analyze the crystal structure, the crystal structure data of the crystal form C of the compound solvate shown in formula (I) is as shown in Table 4, and the crystal structure data confirms the described Crystal form C is a solid form of a solvate of the compound represented by formula (I), and the compound represented by formula (I) in the solvate is combined with 1 molecule of methanol and 1 molecule of water, The specific structure is as follows:

The unimolecular three-dimensional structure ellipsoid diagram of the crystal form C is shown in FIG. 2 .

Table 4

Embodiment 2 Preparation of crystal form A of the compound shown in formula (I)

Dissolve 1.2g of the compound represented by formula (I) in 20mL of ethanol under heating at about 60°C to obtain a hot saturated liquid, filter it hot, cool the filtrate to room temperature at a rate of 5°C/hour, and stir overnight; the precipitated solid It was filtered off and dried under reduced pressure at 40° C. for 16 hours to obtain 0.89 g of a white powder with a purity of 99.6% by HPLC and a yield of 74.2%.

Gained powder sample is the crystalline form A of the compound shown in formula (I), and its X-ray powder diffraction pattern is as shown in Figure 3, and it is expressed in X with 2θ angle, d spacing, peak height percentage and peak area percentage. -In the ray powder diffraction pattern, the 2θ angle, d spacing, peak height percentage and peak area percentage of the main diffraction peaks are shown in Table 5 below, where the characteristic peaks (2θ±0.2°) are 4.839°, 9.141°, 9.694°, 12.433 °, 14.451°, 15.644°, 16.545°, 17.249°, 17.456°, 19.801°, 20.406°, 21.098°, 21.399°, 22.134°, 22.407°, 23.302°, 24.685°, 26.257°, 27.554°, and 27.9

table 5

Its differential scanning calorimetry diagram is shown in Figure 4. DSC test shows that Form A has an endothermic peak at 167.73-170.96°C.

Its thermogravimetric analysis diagram is shown in Figure 5. It can be seen from Figure 5 that the crystal form A loses 0.122% of its weight when heated to 220°C.

Its dynamic moisture adsorption diagram is shown in Fig. 6. The DVS test shows that when the relative humidity changes from 0% to 95%, the mass percentage of the hygroscopic weight gain of the crystal form A is 0.3048%.

Embodiment 3 Preparation of crystal form A of the compound shown in formula (I)

Dissolve 2.0g of the compound represented by formula (I) in 19mL of ethanol under heating at 70-75°C to obtain a hot clear liquid. After stirring for 10 minutes, slowly lower the temperature to 45-55°C. Solids are precipitated. After stirring for 1 hour Add 10 mL of n-heptane dropwise, stir for 1 hour after the dropwise addition, then slowly cool down to room temperature, stir for 2 hours, and filter out the precipitated solid; dry under reduced pressure at 50°C for 16 hours to obtain 1.80 g of white powder, yield 90.0% . The sample obtained by this method is determined to be crystal form A of the compound shown in formula (I) by comparing the X-ray powder diffraction patterns.

Embodiment 4 Preparation of crystal form A of the compound shown in formula (I)

Dissolve 2.0g of the compound represented by formula (I) in 19mL of ethanol under heating at 70-75°C to obtain a hot clear liquid. After stirring for 10 minutes, slowly cool down to 45-50°C. Solids are precipitated. After stirring for 1 hour Add 50 mL of methyl tert-butyl ether dropwise, stir for 1 hour after the dropwise addition, then slowly cool down to room temperature, stir for 2 hours, and filter out the precipitated solid; dry under reduced pressure at 50°C for 16 hours to obtain 1.54 g of white powder, Rate 77.0%. The sample obtained by this method is determined to be crystal form A of the compound shown in formula (I) by comparing the X-ray powder diffraction patterns.

Embodiment 5 Preparation of crystal form A of the compound shown in formula (I)

Disperse 2.0 g of the compound represented by formula (I) in 10 mL of methyl isobutyl ketone, cover and seal it, and then suspend and balance at about 50° C. for 2 days. The solid was filtered off and dried under reduced pressure at 40°C for 16 hours to obtain 1.87 g of white powder, yield: 93.5%. The sample obtained by this method is determined to be crystal form A of the compound shown in formula (I) by comparing the X-ray powder diffraction patterns.

Embodiment 6 Preparation of crystal form B of the compound shown in formula (I)

Dissolve 0.93g of the compound represented by formula (I) in 80mL of methanol under heating at about 60°C to obtain a hot saturated liquid, filter it hot, transfer the filtrate to a 250mL round bottom flask, cover the mouth of the bottle with aluminum foil and make a small hole , slowly evaporate the solvent at room temperature. The solid after the solvent was completely evaporated was dried under reduced pressure at 40° C. for 16 hours to obtain 0.82 g of white powder with a yield of 88.2%.

Gained powder sample is the crystalline form B of the compound shown in formula (I), and its X-ray powder diffraction pattern is as shown in Figure 7, and it is expressed in X with 2θ angle, d distance, peak height percentage and peak area percentage. -In the ray powder diffraction pattern, the 2θ angle, d spacing, peak height percentage and peak area percentage of the main diffraction peaks are shown in Table 6 below, where the characteristic peaks (2θ±0.2°) are 4.958°, 9.838°, 14.718°, 19.631 °, 24.582°, and 31.047°.

Table 6

Its differential scanning calorimetry diagram is shown in Figure 8. DSC test shows that Form B has an endothermic peak at 131.98-142.51°C.

Its thermogravimetric analysis diagram is shown in Figure 9. It can be seen from Figure 9 that the crystal form B loses 0.207% of its weight when heated to 220°C.

Its dynamic moisture adsorption diagram is shown in Fig. 10. The DVS test shows that when the relative humidity changes from 0% to 95%, the mass percentage of the hygroscopic weight gain of the crystal form B is 0.9504%.

Embodiment 7 Preparation of crystal form B of the compound shown in formula (I)

Suspend 0.5 g of the compound represented by the formula (I) in 2 mL of methanol, seal it with a cap, and keep the suspension at room temperature for 2 days. The solid was filtered off and dried under vacuum at 40° C. for 16 hours to obtain 0.37 g of white powder with a yield of 74.0%. The sample obtained by this method is determined to be crystal form B of the compound shown in formula (I) by comparing the X-ray powder diffraction patterns.

Effect Example 1 Stability

1.1 Stability in solution of the crystal form A of the compound shown in formula (I)

Take an appropriate amount of crystal form A samples of the compound represented by formula (I) into a vial, add 1 mL of a single solvent or a mixed solvent into the vial, and ultrasonically disperse to form a suspension. After the suspension was covered and sealed, it was suspended and equilibrated at room temperature for 14 days. Centrifuge, discard the supernatant, and measure the XRPD after drying the solid at 40°C under reduced pressure.

The results showed that in various solvents, the crystal form did not change, and it was still the crystal form A. The single solvent includes ethanol, n-propanol, isopropanol, n-butanol, acetone, 2-butanone, methyl isobutyl ketone, ethyl acetate, isopropyl acetate, isopropyl ether, methyl tert-butyl base ether, acetonitrile, 2-methyltetrahydrofuran, tetrahydrofuran, dichloromethane, toluene, 1,4-dioxane, water, n-heptane, n-hexane, cyclohexane and cyclopentane; the mixed solvent includes The volume ratios of ethanol and water are 15:85, 65:35 and 95:5 respectively, and the volume ratios of isopropanol and water are 35:65, 80:20 and 95:5 respectively.

1.2 The stability of crystal form B of the compound shown in formula (I) in solution

Take an appropriate amount of crystal form B samples of the compound represented by formula (I) into a vial, add 1 mL of a single solvent or a mixed solvent into the vial, and ultrasonically disperse to form a suspension. After the suspension was covered and sealed, it was suspended and equilibrated at room temperature for 14 days. Centrifuge, discard the supernatant, and measure the XRPD after drying the solid at 40°C under reduced pressure.

The results show that in various solvents, the crystal form has not changed, and it is still the crystal form B. The single solvent or mixed solvent includes methanol, methanol and water with a volume ratio of 50:50, 75:25 or 95:5, respectively. , methanol and acetonitrile, methanol and acetone, methanol and ethyl acetate, methanol and isopropyl acetate, methanol and isopropyl ether, methanol and methyl tert-butyl ether, methanol and dichloromethane in a volume ratio of 50:50 .

Effect Example 2 Competition Experiment

Take crystal form A and crystal form B samples of the compound shown in formula (I) of equal weight respectively in the same vial, add 1mL of solvent to the vial, and ultrasonically disperse to form a suspension. The solvent includes ethanol, Acetone, ethyl acetate, and acetonitrile. After the suspension was covered and sealed, it was suspended and equilibrated at room temperature for 7 days. Centrifuge, discard the supernatant, and measure the XRPD after drying the solid at 40°C under reduced pressure.

The results show that the final solid forms are all crystal form A of the compound represented by formula (I), and it can be seen that the crystal form A is a more thermodynamically stable polymorph of the compound represented by formula (I).

Effect Example 3 Stability of crystal form A, crystal form B and amorphous form of the compound represented by formula (I) under high temperature, high humidity and light conditions

Place an appropriate amount of crystal form A or crystal form B of the compound shown in formula (I) or an amorphous test sample in a glass vial, and place it in a glass vial, and heat it under high temperature (60° C.) and high humidity (RH92.5%, 25°C) and light (4500±500Lux, 25°C) and placed open. After 2 weeks, samples were taken and analyzed by X-ray powder diffraction and high performance liquid chromatography (HPLC).

The method of X-ray powder diffraction analysis is the same as described above.

Agilent ZORBAX SB-Phenyl column (3.5μm, 4.6×150mm) was used for reverse-phase HPLC analysis, and the mobile phase components were (A) water containing 0.1% perchloric acid and (B) acetonitrile. The elution gradient was mobile phase B increased from 25% to 95% in 12 minutes, maintained at 95% for 3 minutes, and then equilibrated the system at 25% for 5 minutes. The flow rate was 1.0 mL/min, the injection volume was 2 μL, the column temperature was 25° C., and the detection wavelength was 228 nm. The test article was prepared in methanol to a final concentration of 0.5 mg/mL. Adopt the area normalization method to calculate the purity of total impurity content and the compound shown in formula (I) in need testing sample, the chromatographic peak that peak area is less than 0.05% in need testing product chromatogram is neglected. The inspection results are shown in Table 7-9 below.

Table 7 Influencing factor test results of crystal form A

Table 8 Influencing Factor Test Results of Form B

Table 9 Amorphous Influencing Factors Test Results

The above data in Tables 7-9 show that Form A can increase the purity of the compound represented by formula (I) to about 99.6%, and Form A has a higher active ingredient content than the amorphous form and Form B.

Compared with before the treatment of the influence factor test, the chemical purity and crystal form of the crystal form A and the crystal form B did not change after being placed under high temperature conditions for 2 weeks, and the total impurities did not increase, while the XRPD measurement showed that the The amorphous form has been transformed and its chemical purity has decreased by 2.7%.

Placed under high-humidity conditions for 2 weeks, the crystal forms of Form A, Form B and the amorphous form did not change; the chemical purity of Form A and Form B did not change, and the chemical purity of the amorphous form slightly down.

Placed under light conditions for 2 weeks respectively, the crystal forms of the crystal form A, crystal form B and amorphous form did not change, and the chemical purity of the crystal form A and the crystal form B only decreased by 0.4%-0.5%; The chemical purity of the amorphous form decreased by 3.4%, and the total impurities increased by 3.34%.

It can be seen that, compared with the amorphous solid form, the crystalline form A and crystalline form B of the present invention significantly improve the stability of the compound represented by formula (I) to high temperature and light.

Effect Example 4 Humidity

Using the same DVS assay method as the crystalline form B of the compound shown in formula (I) in embodiment 2, the crystal form B of the compound shown in formula (I) in embodiment 6, as shown in formula (I) The amorphous sample of the compound shown was subjected to DVS determination, and its dynamic moisture adsorption diagram is shown in FIG. 11 . Table 10 summarizes the DVS hygroscopicity results of crystal form A, crystal form B and amorphous form.

Table 10 Crystal Form A, Crystal Form B and the DVS hygroscopicity results of the amorphous form

It can be seen from Table 10 that when the relative humidity changes from 0% to 95%, the weight percentages of moisture absorption weight gain of the crystal form A, crystal form B and amorphous form are 0.3048%, 0.9504% and 4.376% in sequence, and the amorphous form absorbs moisture The mass percentage of weight gain is respectively 14.4 times and 4.6 times that of the crystal form A and the crystal form B. It can be seen that the crystal form A and the crystal form B of the present invention have lower hygroscopicity under high humidity conditions.

It can be seen that compared with the amorphous solid form, the crystal form A and the crystal form B of the present invention significantly improve the hygroscopic performance of the compound represented by formula (I), which is more favorable for the storage of the drug.

Effect Example 5 The in vitro cell activity of the compound shown in formula (I) and its diastereoisomers

CTG method was used to detect the proliferation inhibition experiments of the compounds on NCI-H358, MIA PaCa-2, A549 and A375 cell lines.

NCI-H358 is human non-small cell lung cancer cells with KRas G12C mutation, MIA PaCa-2 is human pancreatic cancer cells with KRasG12C mutation, A549 is human non-small cell lung cancer cells with KRas G12V mutation, A375 is wild-type malignant melanoma cells. The inhibitory effect of the compound on different mutations was evaluated by detecting the inhibitory activity of the compound on the proliferation of four cell lines.

Add 40 μL of the cell suspension to be tested to the 384-well plate (except the peripheral wells) (plate 1: NCI-H358 cell suspension, plate 2: MIA PaCa-2 cell suspension, plate 3: A549 cell suspension, plate 4 : A375 cell suspension). Place the culture plate in a carbon dioxide incubator overnight. The prepared compound was added to each well (by 3-fold dilution, 10 concentration gradient compounds were obtained). Cell plates were incubated for 120 hours in a carbon dioxide incubator. Add 25 μL of CellTiter Glo reagent to the 384-well plate, shake for 10 minutes in the dark, and incubate for 10 minutes. Put the culture plate into EnVision to read the plate, use XLFit to draw the drug efficacy inhibition rate curve and calculate the IC ₅₀ value.

The in vitro cell activity data of the compound represented by formula (I) and its diastereoisomer (Compound 1-1) are shown in Table 11 below.

Table 11 The compound shown in formula (I) and diastereoisomer are to H358 cell, MIA PaCa-2 cell, A549 cell and the proliferation inhibitory activity of A375 cell

The results show that the growth inhibitory activity of the compound shown in formula (I) on H358 cells is 614 times that of its diastereoisomer (compound 1-1), and the growth inhibitory activity on MIA PaCa-2 cells is 326 times of its diastereoisomer (compound 1-1).

The results show that the inhibitory activity of the compound represented by formula (I) on KRas G12C mutant cells is far superior to that of its diastereoisomer (compound 1-1).

Effect Example 6 Pharmacodynamic studies of compounds represented by formula (I) in lung cancer cell NCI-H358 subcutaneous xenograft tumor model

The experimental index is to investigate whether tumor growth can be inhibited, delayed or cured. Tumor diameters were measured twice a week with vernier calipers.

The formula for calculating the tumor volume is: V=0.5×a×b ² , where a and b represent the long diameter and short diameter of the tumor, respectively.

The antitumor efficacy of the test substance was evaluated by T/C (%). T/C%=T _RTV /C _RTV ×100% (T _RTV : RTV of the treatment group; C _RTV : RTV of the solvent control group).

The calculation formula of relative tumor volume RTV is RTV=V _T /V ₁ . Wherein, V ₁ is the tumor volume measured during cage administration (ie Day 1), and V _T is the tumor volume measured on day T.

The percentage value of T/C (%) reflects the curative effect, according to the guiding principle of T/C% ≤ 40% of the anti-tumor drug of the Drug Evaluation Center, it is considered that the test substance is effective. Tumor volume curative effect is evaluated by TGI _V (%), TGI _V (%)=(1-(TV _T-Dn -TV _T-D1 )/(TV _C-Dn -TV _C-D1 ))×100%, TV _C : tumor volume of solvent control group, TV _T : tumor volume of treatment group.

Experiments used BALB/c nude mice. In the experiment, 0.5% MC aqueous solution was added to the crystal form A sample of the compound represented by formula (I), vortexed to make it fully mixed, and suspensions with concentrations of 2.5, 5.0, and 10.0 mg/mL were obtained, respectively. For the administration of 25mg/kg group, 50mg/kg group and 100mg/kg group. Administration by gavage.

The experiment was divided into solvent control group, such as 25mg/kg group, 50mg/kg group and 100mg/kg group of the compound represented by formula (I), administered orally orally for 21 days, once a day. During the administration period, the mice in each group showed good tolerance without obvious abnormalities. See Figure 12 for the change in body weight of the mice.

Figure 13 shows the change in tumor volume in mice. Statistical analysis is carried out to each treatment group and solvent control group tumor volume, as shown in the compound 50mg/kg group and 100mg/kg group shown in formula (I) from the 5th day of administration to the end of administration, there is a very significant difference .

When the test substance began to be administered on the 22nd day, compared with the solvent control group, as the compound 25mg/kg group shown in formula (I), the T/C value of the 50mg/kg group and the 100mg/kg group was 66.58% (TGI %: 42.39%), 36.10% (TGI%: 81.00%) and 13.36% (TGI%: 109.91%).

Experiments show that the 50mg/kg group and 100mg/kg group of the compound represented by formula (I) have significant tumor inhibitory effect in the lung cancer cell NCI-H358 subcutaneous xenograft tumor model.

Effect Example 7 Pharmacodynamic studies of compounds shown in formula (I) in pancreatic cancer cell MIA PaCa-2 subcutaneous xenograft tumor model

Experiments used BALB/c nude mice. In the experiment, 0.5% MC aqueous solution was added to the crystal form A sample of the compound represented by formula (I), vortexed to make it fully mixed, and suspensions with concentrations of 1.5, 3.0, and 10.0 mg/mL were obtained, respectively. For the administration of 15mg/kg group, 30mg/kg group and 100mg/kg group. Administration by gavage.

The experiment was divided into solvent control group, such as 15mg/kg group, 30mg/kg group and 100mg/kg group of the compound represented by formula (I), administered orally orally for 21 days, once a day. During the administration period, the mice in each group showed good tolerance without obvious abnormalities. Figure 14 shows the change in body weight of the mice.

Figure 15 shows the change in tumor volume in mice. Statistical analysis is carried out to each treatment group and solvent control group tumor volume, as the compound 15mg/kg group shown in formula (I), 30mg/kg group and 100mg/kg group, from the 5th day that administration begins to administration end shows There is a very significant difference.

When the compound started to be administered on the 22nd day, compared with the solvent control group, as the compound 15mg/kg group shown in formula (I), the T/C value of the 30mg/kg group and the 100mg/kg group were respectively 36.91% (TGI%: 75.39%), 13.55% (TGI%: 103.21%) and 0.56% (TGI%: 118.78%). The 15mg/kg group, 30mg/kg group and 100mg/kg group of the compound represented by formula (I) showed good anticancer activity.

Experiments have shown that the 15mg/kg group, 30mg/kg group and 100mg/kg group of the compound represented by formula (I) all showed significant tumor inhibitory effects in the pancreatic cancer cell MIA PaCa-2 subcutaneous xenograft tumor model.

Effect Example 8 Pharmacodynamic study of the compound represented by formula (I) in human colon cancer SW837 subcutaneous xenograft tumor model

Experiments used NOD SCID mice. In the experiment, 0.5% MC aqueous solution was added to the crystal form A sample of the compound represented by formula (I), vortexed to mix well, and a suspension with a concentration of 10.0 mg/mL was obtained. Administration by gavage.

The experiment was divided into a solvent control group, such as the 100 mg/kg group of the compound represented by formula (I), which was administered orally or gavaged for 28 days, once a day. During the administration period, the mice in each group showed good tolerance without obvious abnormalities. Figure 16 shows the change in body weight of the mice.

Figure 17 shows the change in tumor volume in mice. Statistical analysis was carried out on the tumor volume of the treatment group and the solvent control group, as shown in the 100 mg/kg group of the compound represented by formula (I), there was a very significant difference from the 4th day of administration to the end of administration.

On the 28th day when the compound was started to be administered, compared with the solvent control group, the T/C value and TGI% value of the 100 mg/kg compound represented by formula (I) were 6.48% and 106.4% respectively. The 100 mg/kg group of the compound represented by the formula (I) showed good anticancer activity.

Experiments show that the 100 mg/kg group of the compound represented by the formula (I) shows a significant tumor-suppressing effect in the human-derived colon cancer SW837 subcutaneous xenograft tumor model.

It should be understood that the examples described herein are for illustrative purposes only, and the examples will help to further understand the present invention, but are not intended to limit the content of the present invention. Many changes in both materials and methods may be made by those skilled in the art without departing from the scope of the invention, and such changes or modifications are included within the spirit and scope of the application and the purview of the appended claims.

Claims

A crystal form A of the compound shown in formula (I), characterized in that its X-ray powder diffraction pattern represented by 2θ angles is at 4.8±0.2°, 12.4±0.2°, 15.6±0.2° and 23.3° There are characteristic peaks at ±0.2°, and the X-ray powder diffraction pattern is measured using the Kα line of the Cu target;
The crystalline form A of the compound shown in formula (I) as claimed in claim 1, is characterized in that, its X-ray powder diffraction figure represented by 2θ angle, at 4.8 ± 0.2 °, 9.1 ± 0.2 °, 9.7 There are characteristic peaks at ±0.2°, 12.4±0.2°, 15.6±0.2°, 19.8±0.2°, 20.4±0.2°, 21.1±0.2°, 23.3±0.2°, 24.7±0.2° and 26.3±0.2°.
The crystal form A of the compound represented by formula (I) as claimed in claim 1, wherein the crystal form A of the compound represented by formula (I) satisfies one or more of the following conditions kind:

(1) Its X-ray powder diffraction pattern represented by 2θ angle, its X-ray powder diffraction pattern represented by 2θ angle, at 4.8±0.2°, 9.1±0.2°, 9.7±0.2°, 12.4±0.2°, 14.5±0.2°, 15.6±0.2°, 16.5±0.2°, 17.2±0.2°, 17.5±0.2°, 19.8±0.2°, 20.4±0.2°, 21.1±0.2°, 21.4±0.2°, 22.1±0.2°, There are characteristic peaks at 22.4±0.2°, 23.3±0.2°, 24.7±0.2°, 26.3±0.2°, 27.6±0.2° and 27.9±0.2°;

(2) The differential scanning calorimetry (DSC) of the crystal form A of the compound represented by formula (I) also has an endothermic peak at 167.7-171.0°C;

(3) In the dynamic moisture sorption diagram (DVS) of the crystal form A of the compound represented by formula (I), the increased weight of the crystal form A is 0%-95% relative to the initial weight. 0.30% weight gain in the humidity range.

The crystalline form A of the compound shown in formula (I) as claimed in claim 3, is characterized in that, its X-ray powder diffraction pattern represented by 2θ angle, its diffraction peak and peak area percentage are shown in the following table:

Numbering 2θ(±0.2°) Peak Area Percentage (%) 1 4.839 100.0 2 9.141 40.6 3 9.694 21.5 4 12.433 37.6 5 14.451 26.0 6 14.644 8.6 7 15.644 84.3 8 16.545 8.4 9 17.249 41.7 10 17.456 9.9 11 19.801 36.8

12 20.406 24.2 13 21.098 48.2 14 21.399 8.9 15 22.134 14.7 16 22.407 4.2 17 23.302 42.3 18 24.685 54.9 19 26.257 30.2 20 27.552 15.8 twenty one 27.948 17.6 twenty two 28.520 2.3 twenty three 29.742 2.2 twenty four 30.746 5.3

.

The crystal form A of the compound represented by formula (I) as claimed in claim 4, wherein the crystal form A of the compound represented by formula (I) satisfies one or more of the following conditions kind:

(1) The X-ray powder diffraction pattern of the crystal form A of the compound shown in formula (I) represented by 2θ angle is basically as shown in Figure 3;

(2) The differential scanning calorimetry (DSC) of the crystal form A of the compound shown in formula (I) is basically as shown in Figure 4;

(3) The thermogravimetric analysis diagram (TGA) of the crystal form A of the compound shown in formula (I) is basically as shown in Figure 5;

(4) The dynamic moisture sorption diagram (DVS) of the crystal form A of the compound represented by formula (I) can also be basically as shown in FIG. 6 .
The preparation method of the crystal form A of the compound represented by formula (I) according to any one of claims 1-5, characterized in that it is method 1, method 2 or method 3:

Method 1: at 40°C to 80°C, cool the hot saturated solution formed by the compound represented by formula (I) and the solvent to room temperature, and crystallize; wherein, the solvent is selected from ethanol, ethyl acetate , one or more of methyl isobutyl ketone, acetonitrile and methyl tert-butyl ether;

Method 2: at 40°C to 80°C, mix the solution formed by the compound represented by formula (I) and solvent A with solvent B, and crystallize; wherein, the solvent A is ethanol, isopropanol, One or more of acetone, ethyl acetate, acetonitrile, tetrahydrofuran, 2-methyltetrahydrofuran and 1,4-dioxane, and the solvent B is selected from methyl tert-butyl ether, n-heptane and water one or more of

Method 3: Place an excess of the compound shown in formula (I) in a solvent to form a suspension. After the suspension is balanced, separate the solid-liquid phase and dry it; wherein, the solvent is selected from ethanol, methyl One or more of isobutyl ketone, methyl tert-butyl ether, acetonitrile, tetrahydrofuran, 2-methyltetrahydrofuran, and 1,4-dioxane.
The preparation method of the crystal form A of the compound shown in formula (I) as claimed in claim 6, wherein the preparation method of the crystal form A of the compound shown in formula (I) satisfies the following conditions One or more of:

(1) In the method 1, method 2 or method 3, the compound shown in formula (I) is a compound shown in formula (I) in the form of an amorphous solid;

(2) In method 1, the volume-mass ratio of the solvent to the compound represented by formula (I) is 10.0-30.0 mL/g;

(3) In method 1, the crystallization is carried out by cooling at a rate of 5-30° C./hour;

(4) In method 1, before carrying out described crystallization, described hot saturated solution is processed through filtration;

(5) In the method 2, the volume-to-mass ratio of the solvent A to the compound represented by formula (I) is 8.0-11.0 mL/g;

(6) In the method 2, the 40°C to 80°C is 70-75°C or 45-50°C;

(7) In the method 2, the volume ratio of the solvent A to the solvent B is (0.2-5):1;

(8) In the method 2, the timing of the mixing is to mix the solution formed by the compound shown in formula (I) and solvent A with solvent B, or mix the compound shown in formula (I) with The solution formed by solvent A is cooled until solids are precipitated and then mixed with solvent B;

(9) In the method 3, the solvent is selected from ethanol and/or methyl isobutyl ketone, for example, methyl isobutyl ketone;

(10) The temperature of the suspension equilibrium is 30°C to 60°C;

(11) In the method 3, the suspension equilibration time is 2 hours to 14 days.
A crystal form B of the compound shown in formula (I), characterized in that its X-ray powder diffraction pattern represented by 2θ angles is at 5.0±0.2°, 9.8±0.2°, 14.7±0.2°, 19.6 There are characteristic peaks at ±0.2°, 24.6±0.2° and 31.0±0.2°, and the X-ray powder diffraction pattern is measured using the Kα line of the Cu target;
The crystal form B of the compound represented by formula (I) as claimed in claim 8, wherein the crystal form B of the compound represented by formula (I) satisfies one or more of the following conditions kind:

(1) Its X-ray powder diffraction pattern represented by 2θ angle, its diffraction peak and peak area percentage are shown in the following table:

Numbering 2θ(±0.2°) Peak Area Percentage (%) 1 4.958 100.0 2 9.838 19.8 3 14.718 1.1 4 19.631 4.6 5 24.582 2.9 6 31.047 1.1

;

(2) The differential scanning calorimetry (DSC) of the crystal form B of the compound represented by formula (I) has an endothermic peak at 132.0-142.5°C;

(3) In the dynamic moisture sorption diagram (DVS) of the crystal form B of the compound represented by formula (I), the increased weight of the crystal form B is relative to the initial weight in the range of 0%-95%. 0.95% weight gain over the humidity range.
The crystal form B of the compound represented by formula (I) as claimed in claim 9, wherein the crystal form B of the compound represented by formula (I) satisfies one or more of the following conditions kind:

(1) The X-ray powder diffraction pattern of the crystal form B of the compound shown in formula (I) represented by 2θ angle is shown in Figure 7;

(2) The differential scanning calorimetry (DSC) of the crystal form B of the compound shown in formula (I) is shown in Figure 8;

(3) The thermogravimetric analysis (TGA) of the crystal form B of the compound shown in formula (I) is shown in Figure 9;

(4) The dynamic moisture adsorption diagram (DVS) of the crystal form A of the compound represented by formula (I) is shown in FIG. 10 .
The preparation method of the crystal form B of the compound represented by formula (I) according to any one of claims 8-10, characterized in that it is method A or method B:

Method A: at 50-70°C, form a hot saturated solution of the compound represented by formula (I) with methanol, and evaporate the solvent at room temperature for crystallization;

Method B: The compound represented by formula (I) is formed into a suspension with a solvent, and after the suspension is balanced, the solid-liquid phase is separated and dried; the solvent is methanol or "a mixture of methanol and water".
The preparation method of the crystal form B of the compound shown in formula (I) as claimed in claim 11, characterized in that, the preparation method of the crystal form B of the compound shown in formula (I) satisfies the following One or more of the conditions:

(1) In the method A or method B, the compound shown in formula (I) is a compound shown in formula (I) in the form of an amorphous solid;

(2) In the method A, before the crystallization step of the volatile solvent, the hot saturated solution is filtered;

(3) In the method A, a post-processing step is also included after the crystallization: drying;

(4) In the method B, when the solvent is a mixture of methanol and water, the volume ratio of methanol to water is (5-95):5;

(5) In the method B, the suspension equilibration time is 2 days to 14 days.
A crystal form C of a compound solvate shown in formula (I), characterized in that the compound solvate shown in formula (I) is monomethanol of the compound shown in formula (I) monohydrate,

The crystal form C belongs to the triclinic crystal system, the space group is P1, and the unit cell parameters are:

α=95.12(3)°, β=93.82(3)°, γ=90.43(3)°; unit cell volume
The number of asymmetric units in the unit cell is Z=1.
A pharmaceutical composition, characterized in that the pharmaceutical composition comprises the crystal form A of the compound shown in formula (I) according to any one of claims 1-5, any one of claims 8-10 One or more of the crystal form B of the compound shown in formula (I) and the crystal form C of the solvate of the compound shown in formula (I) as described in claim 13, and pharmaceutical acceptable excipients.
Application of the compound as shown in formula (I) or the pharmaceutical composition as claimed in claim 14 in the preparation of medicine or Ras inhibitor, said medicine is used for the treatment and/or prevention of Ras-mediated diseases;
Application of the compound as shown in formula (I) or the pharmaceutical composition as claimed in claim 14 in the preparation of medicaments for treating and/or preventing cancer.
The application according to claim 15 or 16, wherein the application meets one or more of the following conditions:

(1) The compound shown in formula (I) is the crystal form A of the compound shown in formula (I) according to any one of claims 1-5, any one of claims 8-10 The crystal form B of the compound represented by formula (I) or the crystal form C of the solvate of the compound represented by formula (I) according to claim 13;

(2) The Ras is a G12C mutation of one or more of K-Ras, H-Ras and N-Ras, such as a G12C mutation of K-Ras;

(3) the disease mediated by Ras is cancer;

(4) The cancer is colon cancer, appendix cancer, pancreatic cancer, MYH-related polyposis, blood cancer, breast cancer, endometrial cancer, gallbladder cancer, bile duct cancer, prostate cancer, lung cancer, brain cancer, ovarian cancer , cervical cancer, testicular cancer, kidney cancer, head or neck cancer, bone cancer, skin cancer, rectal cancer, liver cancer, esophagus cancer, stomach cancer, thyroid cancer, bladder cancer, lymphoma, leukemia and melanoma or Various.