Fused pyridinone salt form, crystalline form and use thereof

Abstract

Provided are a fused pyridinone compound salt type, a crystal form, and applications thereof, and specifically, provided are a salt type of a compound shown in formula (I), a crystal form, and applications thereof.

Description

[0001] The present application claims priority to:

[0002] CN202110321727.X, filing date: 2021 / 03 / 25;

[0003] CN202210273262.X, filing date: 2022 / 03 / 18. TECHNICAL FIELD

[0004] The present application relates to the salt form, crystal form, pharmaceutical composition of the compound represented by formula (I), and the application thereof as KRAS inhibitor. BACKGROUND

[0005] In order to enrich the company's research and development pipeline, focus on unmet medical needs, and innovate drug research and development, it is necessary for the long-term development of the company, and it has important economic and social significance.

[0006] About 3 / 5 of cancer patients have RAS gene mutations. In the study of cancer genes, scientists have found that RAS genes are key genes for cancers including lung cancer, colorectal cancer and pancreatic cancer, etc. more than 20 years ago.

[0007] KRAS mutations account for the absolute majority in pancreatic cancer, colorectal cancer and lung cancer, while NRAS mutations are more common in melanoma and acute myeloid leukemia, and HRAS mutations are more common in bladder cancer and head and neck cancer.

[0008] KRAS gene mutation rate in Asian population is 10-15%, KRAS is one of the main cancer genes that can be mutated in many cancers. KRAS mutant tumors are the most potential targetable molecular subtype of non-small cell lung cancer (NSCLC), with a mutation rate of about 15-25% in non-small cell lung cancer (NSCLC). In cases of NSCLC, KRAS mutations mainly occur at codons 12 and 13. The most common codon variation accounts for about 39% of KRAS mutant NSCLCs, which is KRAS-G12C mutation.

[0009] In lung adenocarcinoma, the positive probability of KRAS gene accounts for 1 / 5-1 / 4, second only to the positive mutation probability of EGFR. The lack of targeted inhibitors makes it very difficult for KRAS-positive non-small cell lung cancer patients in both treatment and prognosis. SUMMARY

[0010] In one aspect of the present application, the present application provides a sulfate salt of the compound represented by formula (I)

[0011] In some embodiments of the present application, the sulfate salt contains 0.5 to 3.0 molar equivalents of sulfuric acid relative to Compound (I).

[0012] In some embodiments of the present application, the sulfate salt contains 0.5 to 2.5 molar equivalents of sulfuric acid relative to Compound (I).

[0013] In some embodiments of the present application, the sulfate salt contains 0.5, 0.6, 0.7, 0.8, 0.9, 1.0, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2.0, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.7, 2.9 or 3.0 molar equivalents of sulfuric acid relative to Compound (I).

[0014] In another aspect of the present application, the present application provides a crystal form A of a sulfate salt of Compound (I). The crystal form A has an X-ray powder diffraction pattern with characteristic peaks at the following 2θ angles: 7.17±0.2°, 11.93±0.2°, 12.34±0.2°, 12.99±0.2°, 16.80±0.2°, 17.75±0.2°.

[0015] In some embodiments of the present application, the crystal form A has an X-ray powder diffraction pattern with characteristic peaks at the following 2θ angles: 7.17±0.2°, 11.93±0.2°, 12.34±0.2°, 12.99±0.2°, 13.54±0.2°, 16.80±0.2°, 17.75±0.2°, 19.09±0.2°, 20.91±0.2°, 21.71±0.2°.

[0016] In some embodiments of the present application, the crystal form A has an X-ray powder diffraction pattern substantially as shown in the X-ray powder diffraction pattern shown in Figure 1

[0017] In some embodiments of the present application, the crystal form A has an X-ray powder diffraction pattern substantially as shown in the X-ray powder diffraction pattern shown in

[0018] Table 1

[0019]

[0020] In some embodiments of the present application, the crystal form A contains 0.5 to 1.5 molar equivalents of sulfuric acid relative to Compound (I).

[0021] In some embodiments of the present application, the crystal form A contains 1.0 to 1.5 molar equivalents of sulfuric acid relative to Compound (I).

[0022] ​In some embodiments of the present application, the above-mentioned crystal form A contains 1.0 to 1.1 molar equivalents of sulfuric acid with respect to compound (I).

[0023] In some embodiments of the present application, the above-mentioned crystal form A contains 0.9, 1.0, 1.1 or 1.2 molar equivalents of sulfuric acid with respect to compound (I). These crystal forms A having different molar equivalents have the same characteristic peaks, for example Figures 13-16 as shown.

[0024] In some embodiments of the present application, the above-mentioned crystal form A contains 1.024, 1.063, 1.075, 1.082, 1.087 or 1.108 molar equivalents of sulfuric acid with respect to compound (I).

[0025] In some embodiments of the present application, the above-mentioned crystal form A is a hydrate, and the water content of the hydrate is 5% to 15%. In some embodiments of the present application, the above-mentioned crystal form A is a hydrate, and the water content of the hydrate is 5.87% to 13.23%.

[0026] In some embodiments of the present application, the above-mentioned crystal form A is a hydrate, and the water content of the hydrate is 6.6% to 10.4%.

[0027] In some embodiments of the present application, the above-mentioned crystal form A is a hydrate, and the water content of the hydrate is 6.6%, 7.0%, 7.5%, 8.0%, 8.5%, 9.0%, 9.5%, 9.6%, 9.7%, 9.8%, 9.9%, 10.0%, 10.1%, 10.2%, 10.3%, 10.4% or 10.5%.

[0028] In some embodiments of the present application, the above-mentioned crystal form A is a hydrate, and the water content of the hydrate is 6.6%, 7.0%, 7.5%, 8.0%, 8.5%, 9.0%, 9.5%, 9.6%, 9.7%, 9.8%, 9.9%, 10.0%, 10.1%, 10.2%, 10.3%, 10.4% or 10.5%, these crystal forms A having different water contents have the same characteristic peaks, for example Figure 1 as shown. 12

[0029] In some embodiments of the present application, the above-mentioned common characteristic peaks are at least 3 peaks selected from the diffraction angle (2θ): 7.17±0.2°, 11.93±0.2°, 12.34±0.2°, 12.99±0.2°, 16.80±0.2°, 17.75±0.2°.

[0030] ​In some embodiments of the present application, the common characteristic peaks are at least 3 peaks selected from the diffraction angles (2θ): 7.17±0.2°, 11.93±0.2°, 12.34±0.2°, 12.99±0.2°, 13.54±0.2°, 16.80±0.2°, 17.75±0.2°, 19.09±0.2°, 20.91±0.2°, 21.71±0.2°.

[0031] In another aspect of the present application, the present application provides a crystal form B of the sulfate salt of the compound represented by formula (I). The crystal form B has characteristic diffraction peaks at the following 2θ angles in the X-ray powder diffraction pattern: 5.67±0.2°, 8.78±0.2°, 10.28±0.2°, 11.02±0.2°, 12.19±0.2°, 15.51±0.2°.

[0032] In some embodiments of the present application, the crystal form B has characteristic diffraction peaks at the following 2θ angles in the X-ray powder diffraction pattern: 5.67±0.2°, 8.78±0.2°, 10.28±0.2°, 11.02±0.2°, 12.19±0.2°, 12.60±0.2°, 13.14±0.2°, 15.51±0.2°, 15.99±0.2°, 19.56±0.2°.

[0033] In some embodiments of the present application, the crystal form B has an X-ray powder diffraction pattern substantially as shown in the X-ray powder diffraction pattern shown in Figure 2 .

[0034] In some embodiments of the present application, the crystal form B has an X-ray powder diffraction pattern substantially as shown in the X-ray powder diffraction pattern shown in

[0035] Table 2

[0036]

[0037] In some embodiments of the present application, the crystal form B contains 0.8-1.5 molar equivalents of sulfuric acid relative to the compound (I).

[0038] In some embodiments of the present application, the crystal form B contains 0.9, 1.0, 1.1 or 1.2 molar equivalents of sulfuric acid relative to the compound (I).

[0039] In another aspect of the present application, the present application provides a crystal form C of the sulfate salt of the compound represented by formula (I). The crystal form C has characteristic diffraction peaks at the following 2θ angles in the X-ray powder diffraction pattern: 5.51±0.2°, 7.71±0.2°, 11.90±0.2°, 13.67±0.2°, 15.69±0.2°, 20.15±0.2°.

[0040] In some embodiments of the present application, the X-ray powder diffraction pattern of the crystalline form C described above has substantially the X-ray powder diffraction pattern as shown in Figure 2. Figure 3

[0041] In some embodiments of the present application, the X-ray powder diffraction pattern of the crystalline form C described above has substantially the X-ray powder diffraction pattern as shown in Figure 2.

[0042] Table 3

[0043]

[0044] In some embodiments of the present application, the crystalline form C described above contains 2.0-3.0 molar equivalents of sulfuric acid relative to compound (I).

[0045] In some embodiments of the present application, the crystalline form C described above contains 2.3, 2.4, 2.5, 2.6 or 2.7 molar equivalents of sulfuric acid relative to compound (I).

[0046] In some embodiments of the present application, the crystalline form C described above is a tetrahydrofuran and ethyl acetate solvate, and the content of tetrahydrofuran in the tetrahydrofuran and ethyl acetate solvate is 0-10 wt%, and the content of ethyl acetate is 0-12 wt%. It should be noted that the crystalline form C can be an ethyl acetate single solvate, i.e. the content of tetrahydrofuran is 0 wt%; or the crystalline form C can be a tetrahydrofuran single solvate, i.e. the content of ethyl acetate is 0 wt%; or the crystalline form C is a mixed tetrahydrofuran and ethyl acetate solvate, i.e. the content of tetrahydrofuran and ethyl acetate is not 0 wt%. In some embodiments of the present application, the crystalline form C described above is a tetrahydrofuran and ethyl acetate solvate, and the content of tetrahydrofuran in the tetrahydrofuran and ethyl acetate solvate is 4.1 wt%, 4.2 wt%, 4.3 wt%, 4.4 wt%, 4.5 wt%, 4.6 wt%, 4.7 wt%, 4.8 wt%, 4.9 wt%, 5.0 wt%, 5.1 wt%, 5.2 wt%, 5.3 wt%, 5.4 wt%, 5.5 wt%, 5.6 wt%, 5.7 wt%, 5.8 wt%, 5.9 wt%, 6.0 wt%, and the content of ethyl acetate is 0.1 wt%.

[0047] In another aspect of the present application, the present application provides a crystalline form D of a sulfate salt of a compound represented by formula (I). The crystalline form D has characteristic diffraction peaks at the following 2θ angles in its X-ray powder diffraction pattern: 6.85±0.2°, 10.29±0.2°, 12.78±0.2°, 16.36±0.2°, 19.84±0.2°, 20.66±0.2°.

[0048] In some embodiments of the present application, the X-ray powder diffraction pattern of the crystalline form D described above has substantially the X-ray powder diffraction pattern as shown in Figure 3. Figure 4 ​X-ray powder diffraction pattern.

[0049] In some embodiments of the present application, the X-ray powder diffraction pattern of the crystalline form D described above has the peak analysis data shown in Table 4 below.

[0050] Table 4

[0051]

[0052] In some embodiments of the present application, the crystalline form D described above contains 0.2 to 0.8 molar equivalents of sulfuric acid with respect to compound (I).

[0053] In some embodiments of the present application, the crystalline form D described above contains 0.3, 0.4, 0.5, or 0.6 molar equivalents of sulfuric acid with respect to compound (I).

[0054] In another aspect of the present application, the present application provides a crystalline form E of a sulfate salt of a compound represented by formula (I). The crystalline form E has an X-ray powder diffraction pattern having characteristic diffraction peaks at the following 2θ angles: 5.52±0.2°, 8.93±0.2°, 11.04±0.2°, 12.16±0.2°, 12.82±0.2°, 22.67±0.2°.

[0055] In some embodiments of the present application, the X-ray powder diffraction pattern of the crystalline form E described above has the peak analysis data shown in Table 5 below.

[0056] In some embodiments of the present application, the X-ray powder diffraction pattern of the crystalline form E described above has substantially the X-ray powder diffraction pattern shown below. Figure 5 X-ray powder diffraction pattern.

[0057] In some embodiments of the present application, the X-ray powder diffraction pattern of the crystalline form E described above has the peak analysis data shown in Table 5 below.

[0058] Table 5

[0059]

[0060] In some embodiments of the present application, the crystalline form E described above contains 0.8 to 1.5 molar equivalents of sulfuric acid with respect to compound (I).

[0061] In some embodiments of the present application, the crystalline form E described above contains 0.9, 1.0, 1.1, or 1.2 molar equivalents of sulfuric acid with respect to compound (I).

[0062] In some embodiments of the present application, the crystalline form E is an ethanol solvate, and the ethanol solvate contains ethanol in an amount of 5% to 22%.

[0063] In some embodiments of the present application, the crystalline form E is an ethanol solvate, and the ethanol solvate contains ethanol in an amount of 17%, 18%, 19%, 20%, 21% or 22%.

[0064] In another aspect of the present application, the present application provides a crystalline form F of a sulfate salt of the compound represented by formula (I). The crystalline form F has an X-ray powder diffraction pattern with characteristic peaks at the following 2θ angles: 5.40±0.2°, 13.01±0.2°, 18.09±0.2°, 21.65±0.2°.

[0065] In some embodiments of the present application, the crystalline form F has an X-ray powder diffraction pattern substantially as shown in the X-ray powder diffraction pattern shown in Figure 6

[0066] In some embodiments of the present application, the crystalline form F has an X-ray powder diffraction pattern substantially as shown in the X-ray powder diffraction pattern shown in

[0067] Table 6

[0068]

[0069] In some embodiments of the present application, the crystalline form F contains sulfate in an amount of 1.0 to 1.5 molar equivalents relative to the compound (I).

[0070] In some embodiments of the present application, the crystalline form F contains sulfate in an amount of 1.1, 1.2, 1.3 or 1.4 molar equivalents relative to the compound (I).

[0071] In some embodiments of the present application, the crystalline form F is a methyl isobutyl ketone and dimethyl sulfoxide solvate, and the methyl isobutyl ketone solvate contains methyl isobutyl ketone in an amount of 0 to 12 wt%, and the dimethyl sulfoxide solvate contains dimethyl sulfoxide in an amount of 0 to 17 wt%. It should be noted that the crystalline form F can be a methyl isobutyl ketone single solvate, i.e. the dimethyl sulfoxide content is 0 wt%, or the crystalline form F can be a dimethyl sulfoxide single solvate, i.e. the methyl isobutyl ketone content is 0 wt%, or the crystalline form F is a mixed methyl isobutyl ketone and dimethyl sulfoxide solvate, i.e. the content of methyl isobutyl ketone and dimethyl sulfoxide is not 0 wt%.

[0072] ​In some embodiments of the application, the crystalline form F described above is a methyl isobutyl ketone and dimethyl sulfoxide solvate, wherein the content of methyl isobutyl ketone in the methyl isobutyl ketone and dimethyl sulfoxide solvate is 0.5 wt%, 0.6 wt%, 0.7 wt%, 0.8 wt%, 0.9 wt%, 1.0 wt%, 1.1 wt%, 1.2 wt%, 1.3 wt%, 1.4 wt% or 1.5 wt%, and the content of dimethyl sulfoxide is 14.0 wt%, 14.1 wt%, 14.2 wt%, 14.3 wt%, 14.4 wt%, 14.5 wt%, 14.6 wt%, 14.7 wt%, 14.8 wt%, 14.9 wt% or 15.0 wt%.

[0073] In another aspect of the application, the application provides a crystalline form G of the sulfate salt of the compound of formula (I). The X-ray powder diffraction pattern of the crystalline form G has characteristic diffraction peaks at the following 2θ angles: 5.88±0.2°, 7.05±0.2°, 9.39±0.2°, 10.87±0.2°, 12.00±0.2°, 16.15±0.2°.

[0074] In some embodiments of the application, the X-ray powder diffraction pattern of the crystalline form G described above has an X-ray powder diffraction pattern substantially as shown in Figure 7. Figure 7

[0075] In some embodiments of the application, the X-ray powder diffraction pattern of the crystalline form G described above has an X-ray powder diffraction pattern substantially as shown in Figure 7.

[0076] Table 7

[0077]

[0078] In another aspect of the application, the application provides a crystalline form H of the sulfate salt of the compound of formula (I). The X-ray powder diffraction pattern of the crystalline form H has characteristic diffraction peaks at the following 2θ angles: 6.11±0.2°, 11.59±0.2°, 12.19±0.2°, 15.66±0.2°, 18.34±0.2°, 13.82±0.2°.

[0079] In some embodiments of the application, the X-ray powder diffraction pattern of the crystalline form H described above has an X-ray powder diffraction pattern substantially as shown in Figure 8. Figure 8

[0080] In some embodiments of the application, the X-ray powder diffraction pattern of the crystalline form H described above has an X-ray powder diffraction pattern substantially as shown in Figure 8.

[0081] Table 8

[0082]

[0083] ​​In some embodiments of the present application, the above-mentioned crystalline form H contains 0.5 to 1.5 molar equivalents of sulfuric acid with respect to compound (I).

[0084] In some embodiments of the present application, the above-mentioned crystalline form H contains 0.9, 1.0, 1.1 or 1.2 molar equivalents of sulfuric acid with respect to compound (I).

[0085] In some embodiments of the present application, the above-mentioned crystalline form H is an anhydrous crystalline form.

[0086] In one aspect of the present application, the present application provides a benzenesulfonic acid salt of a compound represented by formula (I).

[0087] In some embodiments of the present application, the above-mentioned benzenesulfonic acid salt contains 0.5 to 3.0 molar equivalents of benzenesulfonic acid with respect to compound (I).

[0088] In some embodiments of the present application, the above-mentioned benzenesulfonic acid salt contains 1.0 to 2.5 molar equivalents of benzenesulfonic acid with respect to compound (I).

[0089] In some embodiments of the present application, the above-mentioned benzenesulfonic acid salt contains 0.5, 0.6, 0.7, 0.8, 0.9, 1.0, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2.0, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.7, 2.9 or 3.0 molar equivalents of benzenesulfonic acid with respect to compound (I).

[0090] In another aspect of the present application, the present application provides a crystalline form I of a benzenesulfonic acid salt of a compound represented by formula (I). The crystalline form I has an X-ray powder diffraction pattern having characteristic peaks at the following 2θ angles: 7.63±0.2°, 12.05±0.2°, 13.78±0.2°, 15.09±0.2°, 16.04±0.2°, 18.35±0.2°.

[0091] In some embodiments of the present application, the above-mentioned crystalline form I has an X-ray powder diffraction pattern having characteristic peaks at the following 2θ angles: 7.63±0.2°, 11.35±0.2°, 12.05±0.2°, 13.78±0.2°, 15.09±0.2°, 16.04±0.2°, 16.65±0.2°, 18.35±0.2°, 18.87±0.2°, 23.53±0.2°.

[0092] In some embodiments of the present application, the above-mentioned crystalline form I has an X-ray powder diffraction pattern substantially as shown in the X-ray powder diffraction pattern shown in Figure 9

[0093] ​In some embodiments of the present application, the X-ray powder diffraction pattern of the crystalline form I described above is as shown in Table 9.

[0094] Table 9

[0095]

[0096] In some embodiments of the present application, the crystalline form I described above contains 1.5-2.5 molar equivalents of benzene sulfonic acid relative to compound (I).

[0097] In some embodiments of the present application, the crystalline form I described above contains 1.6, 1.7, 1.8, 1.9, 2.0 or 2.1 molar equivalents of sulfuric acid relative to compound (I).

[0098] In another aspect of the present application, the present application also provides a pharmaceutical composition containing the sulfate salt of the compound of formula (I), the benzene sulfonic acid salt of the compound of formula (I) or the crystalline forms A-I of the compound of formula (I) as described above.

[0099] In another aspect of the present application, the present application also provides the use of the sulfate salt of the compound of formula (I), the benzene sulfonic acid salt of the compound of formula (I) or the crystalline forms A-I of the compound of formula (I) as described above or the pharmaceutical composition as described above in the preparation of a medicament for the prevention and / or treatment of KRAS-G12C related diseases.

[0100] In some embodiments of the present application, the KRAS-G12C related diseases described above are selected from non-small cell lung cancer, colon cancer and pancreatic cancer.

[0101] Definitions and explanations

[0102] Unless otherwise defined, all technical and scientific terms used in the present application have the same meanings as commonly understood by one of ordinary skill in the art to which this application belongs. All patents and publications referred to in this application are incorporated herein by reference in their entirety. Although any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present application, the preferred methods, devices, and materials are described.

[0103] "crystalline form" or "crystalline forms" means a solid having a highly regular chemical structure, including, but not limited to, single component or multiple component crystals, and / or polymorphs, solvates, hydrates, clathrates, co-crystals, salts, solvates of salts, hydrates of salts of a compound. Crystalline forms of a substance can be obtained by a number of methods known in the art. Such methods include, but are not limited to, melt crystallization, melt cooling, solvent crystallization, crystallization in confined spaces, e.g., in nanopores or capillaries, crystallization on surfaces or templates, e.g., on polymers, crystallization in the presence of additives such as anti-solvent counter-molecules, desolvation, dehydration, rapid evaporation, rapid cooling, slow cooling, vapor diffusion, sublimation, reactive crystallization, anti-solvent addition, milling, and solvent-drop grinding, and the like.

[0104] "amorphous" or "amorphous form" means a substance in which the particles (molecules, atoms, ions) are arranged in three-dimensional space without periodicity, characterized by a diffuse, peakless X-ray powder diffraction pattern. Amorphous is a special physical form of a solid substance, which has a locally ordered structure characteristic that suggests a close relationship to crystalline substances. Amorphous forms of a substance can be obtained by a number of methods known in the art. Such methods include, but are not limited to, quenching, anti-solvent flocculation, ball milling, spray drying, freeze-drying, wet granulation, and solid dispersion techniques, and the like.

[0105] "solvent" means a substance (typically a liquid) that is capable of completely or partially dissolving another substance (typically a solid). Solvents useful in the practice of the present application include, but are not limited to, water, acetic acid, acetone, acetonitrile, benzene, chloroform, carbon tetrachloride, dichloromethane, dimethyl sulfoxide, 1,4-dioxane, ethanol, ethyl acetate, butanol, t-butanol, N,N-dimethylacetamide, N,N-dimethylformamide, formamide, formic acid, heptane, hexane, isopropanol, methanol, methyl ethyl ketone, l-methyl-2-pyrrolidinone, mesitylene, nitromethane, polyethylene glycol, propanol, 2-propanone, pyridine, tetrahydrofuran, toluene, xylene, mixtures thereof, and the like.

[0106] "anti-solvent" means a fluid that promotes precipitation of a product (or a precursor to a product) from a solvent. An anti-solvent can include a cold gas, or a fluid that promotes precipitation through a chemical reaction, or a fluid that decreases the solubility of a product in a solvent; it can be the same liquid as the solvent but at a different temperature, or it can be a different liquid from the solvent.

[0107] A "solvate" refers to a crystal having a solvent on the surface, or in the lattice, or both, wherein the solvent can be water, acetic acid, acetone, acetonitrile, benzene, chloroform, carbon tetrachloride, dichloromethane, dimethyl sulfoxide, 1,4-dioxane, ethanol, ethyl acetate, butanol, t-butanol, N,N-dimethylacetamide, N,N-dimethylformamide, formamide, formic acid, heptane, hexane, isopropanol, methanol, methyl ethyl ketone, methylpyrrolidinone, mesitylene, nitromethane, polyethylene glycol, propanol, 2-propanone, pyridine, tetrahydrofuran, toluene, xylene, and mixtures thereof, and the like. A specific example of a solvate is a hydrate, wherein the solvent on the surface, or in the lattice, or both, is water. A hydrate can or can not have other solvents in addition to water on the surface, or in the lattice, or both.

[0108] Crystalline or amorphous forms can be identified by a variety of techniques, such as X-ray powder diffraction (XRPD), infrared absorption spectroscopy (IR), melting point method, differential scanning calorimetry (DSC), thermogravimetric analysis (TGA), nuclear magnetic resonance, Raman spectroscopy, X-ray single crystal diffraction, solubility calorimetry, scanning electron microscopy (SEM), quantitative analysis, solubility and dissolution rate, and the like.

[0109] X-ray powder diffraction (XRPD) can detect information of change of crystalline form, crystallinity, crystal state, and the like, and is a common means for identifying crystalline forms. The peak position of an XRPD pattern is mainly dependent on the structure of the crystalline form, and is relatively insensitive to experimental details, while the relative peak height depends on many factors related to sample preparation and instrument geometry. Thus, in some embodiments, the crystalline forms of the present application are characterized by XRPD patterns having certain peak positions, substantially as shown in the XRPD patterns provided in the drawings of the present application. Meanwhile, the measurement of 2Θ of an XRPD pattern can have experimental errors, and the measurement of 2Θ of an XRPD pattern can be slightly different between different instruments and different samples, and thus the values of 2Θ should not be considered as absolute. According to the condition of the instrument used in the experiments of the present application, there is an error tolerance of ±0.2° for the diffraction peak. o

[0110] ​Differential scanning calorimetry (DSC) is a technique that measures the energy difference between a sample and an inert reference (commonly a-Al2O3) as a function of temperature under programmed heating or cooling. The height of the melting peak in a DSC curve depends on many factors related to sample preparation and instrument geometry, while the position of the peak is relatively insensitive to experimental details. Thus, in some embodiments, the crystalline forms described herein are characterized by DSC patterns having characteristic peak positions, as substantially shown in the DSC patterns provided in the figures herein. At the same time, DSC patterns can vary experimentally, and peak positions and peak values can vary slightly between different instruments and different samples, and thus the values of the peak positions or peak values of the DSC endothermic peaks should not be considered absolute. Depending on the instrument conditions used in the experiments described herein, there is a tolerance of ± 3 °C for the melting peak.

[0111] The glass transition refers to the transition between the high-elastic state and the glassy state of an amorphous substance, and is an inherent property of the substance; the transition temperature corresponding to the glass transition is the glass transition temperature (Tg), which is an important physical property of the amorphous substance. The glass transition is a phenomenon related to molecular motion, and thus the glass transition temperature (Tg) mainly depends on the structure of the substance and is relatively insensitive to experimental details. In some embodiments, the amorphous form described herein has a glass transition temperature (Tg) of 107.44 °C, as determined by differential scanning calorimetry (DSC). Depending on the instrument conditions used in the experiments described herein, there is a tolerance of ± 3 °C for the glass transition temperature.

[0112] Differential scanning calorimetry (DSC) can also be used to detect whether the crystalline form has a crystal transformation or a mixed crystal phenomenon.

[0113] Solids with the same chemical composition often form different isomorphs, or variants, with different crystal structures under different thermodynamic conditions, a phenomenon known as polymorphism or homomorphism. When the temperature and pressure conditions change, the variants will transform into each other, a phenomenon known as crystal transformation. Due to crystal transformation, the mechanical, electrical, magnetic, and other properties of the crystal will change greatly. When the temperature of the crystal transformation is within the measurable range, the transformation process can be observed on a differential scanning calorimetry (DSC) graph, which is characterized by an exothermic peak reflecting the transformation process, and at the same time having two or more endothermic peaks, which are the characteristic endothermic peaks of the different crystal forms before and after the transformation. The crystalline form or amorphous form of the compound described herein can undergo crystal transformation under appropriate conditions

[0114] Thermogravimetric analysis (TGA) is a technique that measures the mass of a substance as a function of temperature under programmed control and is suitable for examining the loss of solvent from a crystal or the process of sublimation, decomposition of a sample, and can be used to infer the presence of crystalline water or crystalline solvent in a crystal. The mass change shown in a TGA curve depends on many factors, such as sample preparation and the instrument; there are slight differences in the mass change detected by TGA between different instruments and different samples. In some embodiments, the calcium salt Form A described herein loses about 5.1% of its mass at about 150 °C. Depending on the instrument condition used in the experiment according to the present application, there is an error tolerance of ± 0.3% in the mass change.

[0115] In the context of the present application, the 2-theta values in the X-ray powder diffractograms are given in degrees (°).

[0116] It is noted that the content unit "wt%" and the content unit "%" are used interchangeably and refer to the mass ratio (g / g). For example, in a hydrate, the water content of Form A is 10.02%, which means that the mass ratio (g / g) of water in Form A to the mass of Form A is 10.02. For example, in a solvate, the content of tetrahydrofuran in Form C is 5.1 wt%, which means that the mass ratio (g / g) of tetrahydrofuran in Form C to the mass of Form C is 5.1.

[0117] It is noted that "R" refers to the axial chiral enantiomer.

[0118] When referring to a spectrum or / and data appearing in a figure, "peak" refers to a feature recognizable by a person skilled in the art that cannot be attributed to background noise.

[0119] "Substantially pure" means that a crystal form is substantially free of one or more other crystal forms, i.e., the purity of the crystal form is at least 80%, or at least 85%, or at least 90%, or at least 93%, or at least 95%, or at least 98%, or at least 99%, or at least 99.5%, or at least 99.6%, or at least 99.7%, or at least 99.8%, or at least 99.9%, or the crystal form contains other crystal forms in a percentage of less than 20%, or less than 10%, or less than 5%, or less than 3%, or less than 1%, or less than 0.5%, or less than 0.1%, or less than 0.01% of the total volume or total weight of the crystal form.

[0120] "Substantially free of" means that one or more other crystal forms are in a percentage of less than 20%, or less than 10%, or less than 5%, or less than 4%, or less than 3%, or less than 2%, or less than 1%, or less than 0.5%, or less than 0.1%, or less than 0.01% of the total volume or total weight of the crystal form.

[0121] ​"Relative intensity" means the ratio of the intensity of a peak to the intensity of the first most intense peak in the X-ray powder diffraction pattern (XRPD) when the intensity of the first most intense peak is 100%.

[0122] In the context of the present application, when using or whether using the word "about" or "approximately" or the like, it is meant to indicate that the value or range of values is within 10% of the stated value or range, suitably within 5%, and particularly within 1%. Alternatively, for those skilled in the art, the term "about" or "approximately" means within an acceptable standard deviation of the mean. Whenever a number is disclosed having a plus or minus value, any number within + / - 1%, + / - 2%, + / - 3%, + / - 5%, + / - 7%, + / - 8%, or + / - 10% of the value is also expressly disclosed, where "+" or "-" means plus or minus.

[0123] The term "comprising" is an open term meaning including, but not limited to, whatever follows the term. BRIEF DESCRIPTION OF DRAWINGS

[0124] Figure 1 X-ray powder diffraction (XRPD) pattern of Form A;

[0125] Figure 2 X-ray powder diffraction (XRPD) pattern of Form B;

[0126] Figure 3 X-ray powder diffraction (XRPD) pattern of Form C;

[0127] Figure 4 X-ray powder diffraction (XRPD) pattern of Form D;

[0128] Figure 5 X-ray powder diffraction (XRPD) pattern of Form E;

[0129] Figure 6 X-ray powder diffraction (XRPD) pattern of Form F;

[0130] Figure 7 X-ray powder diffraction (XRPD) pattern of Form G;

[0131] Figure 8 X-ray powder diffraction (XRPD) pattern of Form H;

[0132] Figure 9 X-ray powder diffraction (XRPD) pattern of Form I;

[0133] Figure 10 Graph of NCI-H358 cell seeding day versus body weight change following administration of the compound of formula (I);

[0134] Figure 11 Figure 6 is a plot of NCI-H358 cell inoculation day versus tumor volume for dosing of Compound (I);

[0135] Figure 12 Figure 7 is an X-ray powder diffraction (XRPD) pattern of Form A with different water content;

[0136] Figure 13 Figure 8 is an X-ray powder diffraction (XRPD) pattern of Sample X-1;

[0137] Figure 14 Figure 9 is an X-ray powder diffraction (XRPD) pattern of Sample X-2;

[0138] Figure 15 Figure 10 is an X-ray powder diffraction (XRPD) pattern of Sample X-3;

[0139] Figure 16 Figure 11 is an X-ray powder diffraction (XRPD) pattern of Sample X-4. DETAILED DESCRIPTION

[0140] The present application is described in detail below by way of Examples, but it is not meant to present any adverse limitations on the present application. The present application has been described in detail and specific embodiments thereof have been disclosed with particularity, but it is to be understood that various changes and modifications can be suggested to one skilled in the art, and it is intended that the present application encompass such changes and modifications as fall within the spirit and scope of the application.

[0141] XRPD patterns were collected on a PANalytical X'Pert Pro X-ray powder diffractometer with the following scan parameters as shown in Table 10.

[0142] Table 10

[0143]

[0144] TGA and DSC patterns were collected on a TA Q5000 / Discovery 5500 thermogravimetric analyzer and a TA Q2000 / Discovery 2500 differential scanning calorimeter, respectively. The test parameters are listed in Table 11.

[0145] Table 11

[0146] Parameters TGA DSC mDSC Method Linear Ramp Linear Ramp Modulated Ramp Sample Pan Aluminum pan, open Aluminum pan, crimped / uncrimped Aluminum pan, crimped Temperature Range Room temperature - set endpoint temperature 25 °C - set endpoint temperature 25 °C - set endpoint temperature Scan Rate (°C / min) 10 10 3 Protective Gas Nitrogen Nitrogen Nitrogen

[0147] Dynamic vapor sorption (DVS) curves were collected on a SMS (Surface Measurement Systems) DVS Intrinsic. The relative humidity at 25 °C was corrected using the deliquescence points of LiCl, Mg(N03)2, and KCl. The DVS test parameters are listed in Table 12.

[0148] Table 12

[0149]

[0150] The liquid nuclear magnetic spectrum of the present application was collected on a Bruker 400M nuclear magnetic resonance instrument, with DMSO-d6 as the solvent.

[0151] The PLM image of the present application was taken by an Olympus SZX7 stereomicroscope.

[0152] The dissociation constant of the compound of the present application was tested by Sirius pKa log P / D tester (model: T3Dt).

[0153] In the test of the present application, the purity test, dynamic solubility and stability test were tested by Agilent 1260 high performance liquid chromatograph, and the ion salt-forming molar ratio test was tested by ion chromatography, and the analysis conditions are shown in Table 13 and Table 14.

[0154] Table 13

[0155]

[0156]

[0157] Table 14

[0158] Ion Chromatograph ThermoFisher ICS-1100 Chromatographic Column IonPac AS18 Analytical Column, 250*4mm Mobile Phase 25 mM NaOH Injection Volume 25 μL Flow Rate 1.0 mL / min Temperature 35℃ Column Temperature 35℃ Current 80 mA Run Time SO4 2- : 13.0 min

[0159] The moisture test method (KF method): the present application adopts volumetric method to determine the moisture in the sample, and the parameters are shown in Table 15 below. The moisture determination based on the Karl Fischer method may produce an error within ±0.3%. Therefore, the value of the moisture content should be understood as also including values within the range of ±0.3%.

[0160] Table 15

[0161] Instrument Karl Fischer Water Titrator, Model 852 (Mettler Toledo) Titrant Volumetric Titration Reagent (Honeywell-Fluka) Solvent Anhydrous Methanol (Scharlau)

[0162] The solvent abbreviations or Chinese meanings of English used in the present application are shown in Table 16 below:

[0163] Table 16

[0164] English Chinese English Chinese MeOH Methanol 1,4-Dioxane 1,4-Dioxane EtOH Ethanol ACN Acetonitrile IPA Isopropyl Alcohol DCM Methylene Chloride Acetone Acetone CHCl3 Trichloromethane MIBK Methyl Isobutyl Ketone Toluene Toluene EtOAc Ethyl Acetate n-Heptane n-Heptane IPAc Isopropyl Acetate DMSO Dimethyl Sulfoxide MTBE Methyl Tert-Butyl Ether Anisole Anisole THF Tetrahydrofuran CPME Cyclopentyl Methyl Ether 2-MeTHF 2-Methyl Tetrahydrofuran [H2O] Water

[0165] The present application discloses the salt type, crystal form of the compound of formula (I) and the preparation method thereof. The skilled in the art can refer to the content of the present application, and appropriately improve the process parameters to realize. It is particularly pointed out that all similar substitutions and changes are obvious to the skilled in the art, and they are considered to be included in the present application. The method of the present application has been described by the preferred embodiment, and the relevant personnel can obviously modify or appropriately change and combine the method described herein without departing from the content, spirit and scope of the present application, to realize and apply the present application technology.

[0166] In order to further understand the present application, the present application is described in detail below in combination with examples.

[0167] Example 1: Preparation of the compound of formula (I)

[0168] Step 1: Preparation of compound 8-2

[0169]

[0170] The raw material 8-1 (10 g, 52.351 mmol) was dissolved in thionyl chloride (30 mL), and the system was heated to 85°C for 16h. The system was concentrated, and the residue was dissolved in 1,4 dioxane (30 mL), and the solution was slowly added to the stirred methanol at 0°C, and the system was heated to 70°C for 2h. The system was concentrated to give compound 8-2.

[0171] Step 2: Preparation of compound 8-3

[0172]

[0173] Compound 8-2 (4 g, 19.4 mmol) was dissolved in methanol (50 mL), and sodium methoxide in methanol (4 mL, 21.3 mmol) was added dropwise, and the system was reacted at room temperature (20°C) for 3h. The system was concentrated, poured into water (50 mL), extracted with ethyl acetate (50 mL x 3), and the combined organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to give crude 8-3.

[0174] MS (ESI) m / z (M+H) + = 202.0.

[0175] Step 3: Preparation of compound 8-4

[0176]

[0177] Compound 8-3 (1.48 g, 7.36 mmol), compound 3-9 (1.11 g, 7.36 mmol), palladium acetate (165 mg, 0.736 mmol), 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene (425 mg, 0.735 mmol), cesium carbonate (4.8 g, 14.73 mmol) were dissolved in dioxane (15 mL) under nitrogen atmosphere. The system was heated to 110 °C and stirred for 4 h. The system was cooled to room temperature and concentrated to give a crude product. The crude product was purified by column chromatography (ethyl acetate / petroleum ether (v / v) = 0-20%) to give compound 8-4.

[0178] MS (ESI) m / z (M+H) + = 316.0.

[0179] Step 4: Preparation of compound 8-5

[0180]

[0181] Compound 8-4 (1.58 g, 4.80 mmol) was dissolved in N,N-dimethylformamide (15 mL), and N-chlorosuccinimide (0.706 g, 5.28 mmol) was added thereto. The system was heated to 80 °C and reacted for 5 h. The system was cooled to room temperature and poured into water (50 mL), extracted with ethyl acetate (50 mL x 3), and the combined organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to give a crude product. The crude product was purified by column chromatography (ethyl acetate / petroleum ether (v / v) = 0-5%) to give compound 8-5.

[0182] MS (ESI) m / z (M+H) + = 350.0

[0183] Step 5: Preparation of compound 8-6

[0184]

[0185] Compound 8-5 (6.3 g, 7.82 mmol) was dissolved in N,N-dimethylformamide (30 mL), and sodium hydride (2.17 g, 54.15 mmol) was added portionwise at 0 °C. After the addition was completed, the system was reacted at 0 °C for 30 min, and acetyl chloride (3.85 mL, 54.15 mmol) was added dropwise. Water (30 mL) and a saturated aqueous potassium carbonate solution were sequentially added to the system, which was reacted at room temperature (20 °C) for 3 h. After extraction with EA (100 mL x 2), the aqueous phase was adjusted to pH 4-5 with hydrochloric acid (4 N) and extracted with ethyl acetate (100 mL x 4). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to give a crude product, which was purified by medium pressure column chromatography (methanol / dichloromethane (v / v) = 0-5%) to give compound 8-6.

[0186] MS (ESI) m / z (M+H) + = 360.0.

[0187] Step 6: Preparation of compound 8-7

[0188]

[0189] Compound 8-6 (1.86 g, 5.18 mmol) was dissolved in glacial acetic acid (30 mL), and nitric acid (15 mL) was added dropwise to the system at room temperature (20 °C). After the addition was completed, the system was stirred at room temperature (20 °C) for 2 h. The system was concentrated to remove most of the glacial acetic acid, and the residue was poured into ice water (25 mL), adjusted to pH 5-6, filtered, and the filter cake was dried after water washing to give compound 8-7.

[0190] MS (ESI) m / z (M+H) + = 405.0.

[0191] Step 7: Preparation of compound 8-8

[0192]

[0193] Compound 8-7 (1 g, 2.47 mmol) was dissolved in a mixed solution of acetic acid (6 mL) and hydrobromic acid (8 mL). The system was warmed to 100 °C and reacted for 16 h. The system was rotary evaporated to give compound 8-8.

[0194] MS (ESI) m / z (M+H) + = 391.0.

[0195] Step 8: Preparation of compound 8-9

[0196]

[0197] Compound 8-8 (2.0 g, 5.13 mmol) and N,N-diisopropyl ethylamine (5 mL, 30.7 mmol) were dissolved in acetonitrile (6 mL), and to the solution was added phosphorus oxychloride (7 mL, 77 mmol) at room temperature (20 °C). After the addition, the system was warmed to 80 °C and stirred for 2 h. The system was concentrated to give a crude product. The crude product was purified by column chromatography (ethyl acetate / petroleum ether (v / v) = 0~50%) to give compound 8-9.

[0198] MS (ESI) m / z (M+H) + = 427.0.

[0199] Step 9: Preparation of compound 25-1

[0200]

[0201] Compound 8-9 (426 mg, 1.0 mmol), 7-1 (286 mg, 1.1 mmol), N,N-diisopropyl ethylamine (0.2 mL) were dissolved in acetonitrile (10 mL), and the system was warmed to 100 °C and stirred for 4 h. The system was concentrated to give a crude product. The crude product was purified by column chromatography (ethyl acetate / petroleum ether (v / v) = 0~35%) to give compound 25-1.

[0202] MS (ESI) m / z (M+H) + = 649.0.

[0203] Step 10: Preparation of compound 25-2

[0204]

[0205] Compound 25-1 (326 mg, 0.502 mmol), iron powder (200 mg, 3.6 mmol) were dissolved in acetic acid (15 mL), and the system was warmed to 85 °C and stirred for 1 h under nitrogen atmosphere. The system was filtered through diatomite, and the filtrate was concentrated. The residue was dissolved in ethyl acetate and washed with saturated sodium bicarbonate. The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to give compound 25-2. It was used in the next reaction without further purification. MS (ESI) m / z (M+H) + = 587.0.

[0206] Step 11: Preparation of compound 25-3

[0207]

[0208] Compound 25-2 (277 mg, 0.5 mmol), compound 2-3 (282 mg, 1 mmol), tetrakis(triphenylphosphine)palladium (150 mg, 0.125 mmol), potassium carbonate (138 mg, 1 mmol) were dissolved in a mixed solution of dioxane (18 mL) and water (1.8 mL). The system was heated to 100°C under nitrogen atmosphere and stirred for 2 h. The system was concentrated to obtain a crude product, which was purified by medium pressure column chromatography (ethyl acetate / petroleum ether (v / v) = 0-50%) to obtain compound 25-3.

[0209] MS (ESI) m / z (M+H) + = 707.2.

[0210] Step 12: Preparation of compound 29-1

[0211]

[0212] Compound 25-3 (700 mg, 1 mmol) and cesium carbonate (977 mg, 3 mmol) were dissolved in N,N-dimethylformamide (20 mL), and compound 26-1 (432 mg, 3 mmol) was added thereto at room temperature (25°C). After the addition, the system was heated to 120°C under nitrogen atmosphere and stirred for 2 h. The system was filtered, and the filtrate was concentrated to obtain a crude product, which was purified by silica gel column chromatography (dichloromethane / methanol (v / v) = 1 / 10) to obtain compound 29-1.

[0213] MS (ESI) m / z (M+H) + = 778.2.

[0214] Step 13: Preparation of compound 29-2

[0215]

[0216] Compound 29-1 (150 mg, 0.2 mmol), hydrochloric acid (6N, 7 mL) were added to a mixed solution of methanol (0.6 mL) and tetrahydrofuran (6 mL). The system was heated to 55°C and reacted for 10 min. The system was concentrated to obtain a crude product of compound 29-2, which was used in the next reaction without further purification.

[0217] MS (ESI) m / z (M+H) + = 634.2.

[0218] Step 14: Preparation of compound 29

[0219]

[0220] Compound 29-2 (140 mg, 0.2 mmol) was dissolved in dichloromethane (10 mL), the system was cooled to 0 °C, and triethylamine (0.3 mL, 2.1 mmol) and acryloyl chloride (27 mg, 0.3 mmol) were added dropwise. The system was reacted at 0 °C for 0.5 h. After the system was quenched with methanol, the crude product was concentrated. The crude product was dissolved in methanol (5 mL), and potassium carbonate (140 mg) was added. After stirring at room temperature (20 °C) for 30 min, the system was adjusted to pH ~ 6 with hydrochloric acid, and extracted with dichloromethane (20 mL) and water (20 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain the crude product. The crude product was purified by high-performance preparative liquid chromatography (separation conditions: column Welch C1821.2 x 250 mm, 10 μm; column temperature: 25 °C; mobile phase: water (10 mM / L NH4HCO3) - acetonitrile; acetonitrile 40% - 60% 9 min; flow rate 30 mL / min) to obtain compound 29.

[0221] MS (ESI) m / z (M+H) + = 688.2.

[0222] Step 15: Preparation of the compound of formula (I)

[0223]

[0224] The diastereomeric compound 29 was purified by SFC (ChiralPak AD, 250 x 30 mm I.D., 10 μm; mobile phase: [CO2-ethanol (0.1% ammonia water)]; ethanol%: 25%; flow rate: 60 mL / min; column temperature: 38 °C). After concentration, the compound of formula (I) was obtained.

[0225] The compound of formula (I):

[0226] 1H NMR (400 MHz, DMSO-d6) δ 10.18 (br s, 1H), 8.45 (d, J = 4.9 Hz, 1H), 8.26 (s, 1H), 7.29 - 7.20 (m, 2H), 7.04 (dd, J = 16.8, 10.4 Hz, 0.75H), 6.86 (dd, J = 17.6, 10.4 Hz, 0.25H), 6.72 - 6.60 (m, 2H), 6.14 (d, J = 16.4 Hz, 1H), 5.75 (d, J = 10.7 Hz, 1H), 5.03 (d, J = 13.8 Hz, 0.25H), 4.80 (d, J = 7.8 Hz, 0.75H), 4.61 (d, J = 14.1 Hz, 1H), 4.43 - 4.30 (m, 1H), 4.28 - 4.15 (m, 1H), 4.04 - 3.89 (m, 1H), 3.75 (dd, J = 14.5, 4.4 Hz, 1H), 3.28 - 3.10 (m, 2H), 2.75 - 2.65 (m, 1H), 2.39 - 2.28 (m, 1H), 2.28 - 2.17 (m, 1H), 2.06 - 1.96 (m, 6H), 1.81 (d, J = 9.5 Hz, 3H), 1.53 (d, J = 6.8 Hz, 3H), 1.11 (d, J = 6.9 Hz, 3H), 0.95 (d, J = 6.6 Hz, 3H).

[0227] MS (ESI) m / z (M+H) + = 688.3.

[0228] HPLC retention time 5.269 min.

[0229] Separation conditions: Column: Waters XBridge 4.6*100mm, 3.5um; Column temperature: 40 °C; Mobile phase: water (10 mM ammonium bicarbonate) - acetonitrile; acetonitrile: 5% - 95% 7 min; Flow rate: 1.2 mL / min. SFC 100% ee. Retention time 4.349 min.

[0230] Separation conditions: Column: ChiralPak AD-3, 150 x 4.6 mm I.D., 3 μm; Mobile phase: [CO2 - Ethanol (0.05% DEA)]; Ethanol %: 5% - 40% 5 min, 40% 2.5 min, 5% 2.5 min; Flow rate: 2.5 mL / min; Column temperature: 35 °C.

[0231] Example 2: Preparation and characterization of Form A

[0232] Into a 20-mL glass bottle was placed 500.2 mg of the free starting material (compound of Formula I) and 4 mL of THF / toluene (1 : 1, v / v) was added. Into a 20-mL glass bottle was placed 180 μL of 4 M sulfuric acid and 6 mL of THF / toluene (1 : 1, v / v) was added to dilute. The diluted sulfuric acid solution was added dropwise to the free starting material with magnetic stirring (-750 rpm). After stirring at room temperature for about 1 day, the resulting solid was isolated by suction filtration and dried under vacuum at room temperature for 2 days. The solid was collected.

[0233] The XRPD structure is shown in Figure 1

[0234] The water content of Form A was determined by KF test, and the results are shown in Table 17. The water content of the sample was 10.0% (the theoretical crystalline water content of the pentahydrate is about 10.3%).

[0235] Table 17

[0236]

[0237] Notes: * The average titer used in the titration was 4.7976 mg / mL.

[0238] Two more batches of Form A were prepared in the same manner and were identified as Form A. The water content of the samples was determined and the results are shown in Table 18.

[0239] Table 18

[0240] Sample XPRD Pattern Water content (%) * ]] 1 Figure 12 (sample 1) 10.4％ 2 Figure 12 (sample 2) 6.6％

[0241] Sulfuric acid content determination method:

[0242] Instrumentation:

[0243] Ion chromatograph, electronic balance of 100,000th, ultrasonic instrument.

[0244] Reagents and reference substances:

[0245] Table 19

[0246] Name Grade Water Purified Water Potassium Sulfate Analytical Reagent

[0247] Chromatographic conditions:

[0248] Table 20

[0249] Name Parameters Chromatographic Column Dionex IonPac AS11-HC, 4*250mm, 13 μm Guard Column Dionex IonPac AG11-HC, 4*50mm, 13 μm Eluent 30 mM KOH Flow Rate 1.3 ml / min Run Time 12 min Injection Volume 25 μl Column Temperature 35℃ Suppressor Model AERS_4mm

[0250] Solution preparation:

[0251] Diluent / blank solution: water.​

[0252] Stock solution of reference (0.5 mg / ml): Accurately weigh about 50 mg of potassium sulfate into a 100-ml volumetric flask, dissolve in diluent and dilute to the mark, mix well.

[0253] Solution of reference (0.05 mg / ml): Pipette 1 ml of stock solution of reference into a 10-ml volumetric flask, dilute to the mark with diluent, mix well.

[0254] Control solution: Prepared as solution of reference.

[0255] Solution of test (0.2 mg / ml): Accurately weigh about 20 mg of test sample into a 100-ml volumetric flask, dilute to the mark with diluent, mix well. Prepare in duplicate.

[0256] Injection sequence:

[0257] After system equilibration, inject in the following sequence, inject one injection of reference solution after every 12 injections of test solution or at the end of the sequence.

[0258] Table 21

[0259] Solution Name Injection Needle Number Blank Solution ≥1 Reference Solution 5 Control Solution 1 Test Solution 1 1 Test Solution 2 1 (Back In) Reference Solution 1

[0260] System suitability:

[0261] Blank interference: The blank solution should have no interference (if there is interference, it should not exceed 0.2% of the peak area of the main peak of the first injection of reference solution)

[0262] Repeatability: The reference solution is injected 5 times in succession, the RSD of the peak area of the main peak should be ≤ 5%.

[0263] Recovery: The recovery of the peak area of the main peak in the reference solution and the control solution should be between 95% and 105%. The recovery of the peak area of the main peak in the reference solution injected back should be between 95% and 105%.

[0264]

[0265] A CS : The peak area of the target in the control solution

[0266] C RS : The concentration of the target in the reference solution, mg / ml

[0267] The average of the peak area of the target of 5 injections of the reference solution in succession

[0268] C CS : The concentration of the target in the control solution, mg / ml

[0269]

[0270] A RS : Target peak area in the control solution

[0271] Average of target peak area in the control solution for 5 consecutive injections

[0272] Result calculation

[0273]

[0274] A SPL : Target peak area in the test solution

[0275] C RS : Concentration of target in the control solution, mg / ml

[0276] P: Content of the control, %

[0277] Average of target peak area in the control solution for 5 consecutive injections

[0278] C SPL : Concentration of the test solution, mg / ml

[0279]

[0280] m: Molecular weight of sulfuric acid (98)

[0281] n: Molecular weight of sulfate (96)

[0282] Reporting mode

[0283] Results are reported as averages, with 4 significant figures;

[0284] The relative deviation of two results should not be more than 2.0%.

[0285] The above method was used to identify another four batches of crystal form, and it was found that they were all crystal form A. The sulfuric acid content was determined, and the results are shown in Table 22.

[0286] Table 22

[0287] Sample XPRD Pattern Free Base (Compound of Formula I) Content % Sulfuric Acid Content % Molar Ratio of Sulfuric Acid:Free Base X-1 Figure 13 77.69 11.98 1：1.082 X-2 Figure 14 76.92 11.91 1：1.087 X-3 Figure 15 76.21 12.03 1：1.108 X-4 Figure 16 76.00 11.64 1：1.075

[0288] Note: The molar mass of free base is 688.20 g / moL, and the molar mass of sulfuric acid is 98.07 g / moL.

[0289] Example 3: Solid state stability experiment of crystal form A

[0290] The physical and chemical stability of the sulfate Form A sample and the free sample (Formula I compound) were tested by XRPD and HPLC after the samples were left open at 25°C / 60% RH and 40°C / 75% RH for 1 week and 2 weeks. Meanwhile, the stability of the sulfate Form A sample was further tested by leaving the sample open at 25°C / 60% RH and 40°C / 75% RH for 1 month and closed at 60°C for 24 hours. The results of the stability test are summarized in Table 23. The results show that no change in crystal form was observed for the sulfate Form A sample under the test conditions, and no significant decrease in HPLC purity was observed. The free sample was still amorphous after the stability test, and a decrease in HPLC purity of about 1.5-3.9 area% was observed under the test conditions.

[0291] Table 23

[0292]

[0293] Example 4: Preparation and identification of Form B

[0294] Sulfate Form B was obtained by suspending and stirring sulfate Form A in a MeOH solvent system at room temperature for about 8 days, and then drying the solid sample at room temperature for about 1 day.

[0295] The XRPD structure is shown in Figure 4. Figure 2

[0296] Example 5: Preparation and identification of Form C

[0297] Sulfate Form C was obtained by suspending and stirring a 4M solution of sulfuric acid and the starting free sample (Formula I compound) in a molar ratio of 2.5:1 in THF at room temperature for about 4 days, centrifuging the solid sample, and then drying the sample at room temperature under vacuum for about 8 hours, and then using EtOAc to spin and centrifuge the sample three times.

[0298] The XRPD structure is shown in Figure 5. The molar ratio of acid:free form in the sample was 2.5, as tested by HPLC / IC. Figure 3

[0299] Example 6: Preparation and identification of Form D

[0300] Sulfate Form D was obtained by suspending and stirring a 4M solution of sulfuric acid and the starting free sample (Formula I compound) in a molar ratio of 0.5:1 in THF at room temperature for about 4 days, centrifuging the solid sample, and then drying the sample at room temperature under vacuum for about 8 hours, and then testing the XRPD, to obtain an amorphous sample. The amorphous sample was then suspended and stirred in ACN:H2O (19:1, v:v) at room temperature for about 2 days to obtain Form D.

[0301] The XRPD structure is shown in Figure 6. The molar ratio of acid:free form in the sample was 0.5, as tested by HPLC / IC. Figure 4 ​​​

[0302] Example 7: Preparation and identification of Form E

[0303] Form E was obtained by slow evaporation of a sample of sulfate Form A in EtOH / DCM (4:1, v / v) system.

[0304] The XRPD structure is shown in Figure 5 .

[0305] Example 8: Preparation and identification of Form F

[0306] Form F was obtained by stirring a sample of sulfate Form A in DMSO:MIBK (1:19, v:v) solvent system at room temperature for about 5 days.

[0307] The XRPD structure is shown in Figure 6 . The sample was tested by HPLC / IC, and the acid:free molar ratio was 1.3.

[0308] Example 9: Preparation and identification of Form G

[0309] Form G sample of sulfate was obtained by antisolvent addition of starting free sample (compound of Formula I) in EtOH / n-Heptane system.

[0310] The XRPD structure is shown in Figure 7 .

[0311] Example 10: Preparation and identification of Form H

[0312] Form H was obtained by dehydration of sulfate Form A at high temperature under nitrogen protection and cooling to 30°C.

[0313] The XRPD structure is shown in Figure 8 .

[0314] Example 11: Preparation and identification of Form I

[0315] Form I was obtained by stirring a sample of free starting sample (compound of Formula I) and benzene sulfonic acid at a molar ratio of 1:2 in EtOAc solvent system at room temperature for about 2 days, centrifuging the solid sample and drying it at room temperature under vacuum for 17 hours, and then stirring it in EtOAc solvent system at room temperature for about 4 days, and then drying the separated solid at room temperature under vacuum for about 20 hours.

[0316] The XRPD structure is shown in Figure 9 .

[0317] Effect example:

[0318] Experimental example 1: Inhibition of RAS-mediated signal transduction

[0319] The ability of the compounds disclosed herein to inhibit RAS-mediated signaling was assessed and demonstrated as follows. Cells NCI-H358 (ATCC Catalog No. CRL-5807) expressing mutant RAS (G12C) were cultured with RPMI medium containing 10% fetal bovine serum, penicillin / streptomycin double antibiotic. Cells were plated at 40,000 cells per well in a 96-well plate (Corning Catalog No. 3699) and allowed to adhere to the plate bottom overnight. Cells were treated with the compounds of the present application or without the compounds of the present application (dimethyl sulfoxide, DMSO) and the final concentration of DMSO was ensured to be 0.5%. After 2 hours of treatment, the medium was removed and 4% paraformaldehyde (Beyotime Catalog No. E672002-0100) was added and allowed to stand for 20 minutes. After cell fixation, PBS was used to wash and pre-cooled methanol was used to incubate for 10 minutes to permeabilize the cell membrane. 1X blocking buffer (Thermo Catalog No. 37520) was added to block the binding of non-specific antibodies for 1 hour.

[0320] Detection of phosphorylated ERK level was performed using enzyme-linked immunosorbent assay (ELISA) method. Phosphorylated ERK antibody (Cell Signal Technology Catalog No. 4370) was diluted at 1:400 with 0.05% Tween 20 containing 1X blocking buffer, added to the 96-well plate and incubated at 4°C overnight. The plate was washed 5 times with PBS containing 0.05% Tween 20. HRP-conjugated secondary antibody (Thermo Catalog No. 31460) was diluted at 1:10,000 with 0.05% Tween 20 containing 1X blocking buffer, added to the 96-well plate and incubated at room temperature for 2 hours. The plate was washed 5 times with PBS containing 0.05% Tween and TMB (Thermo Catalog No. 4816) was added and incubated at room temperature for 15 minutes. The reaction was stopped by adding 1 mol / L H2SO4and OD value was read at 450 nm wavelength using EnVision (PerkinElmer).

[0321] Detection of total cell number per well was performed using Janus green staining method. The 96-well plate after detection of phosphorylated ERK level was washed with PBS until colorless and 0.1% Janus green (Abeam Catalog No. ablll1622) was added and incubated for 10 minutes. After washing with double distilled water, 0.1 mol / L HC1 was added and incubated for 10 minutes with shaking. OD value was read at 595 nm wavelength using EnVision (PerkinElmer).

[0322] The signal of pERK (Thr202 / Tyr204) was normalized using the signal value of Janus green and the percentage of inhibition relative to DMSO reference after drug treatment was calculated. The percentage value was fitted by four-parameter dose response curve and IC50 value was generated. The experimental results are shown in Table 24.

[0323] Table 24

[0324] Compound Number p-ERK IC50 (NCI H358, μM) ARS-1620 0.325 Compound of Formula (I) 0.009

[0325] The compounds of the present application exhibit excellent ability to inhibit RAS-mediated signaling.

[0326] Experimental Example 2: Growth inhibition ability experiment of tumor cell lines expressing KRAS-G12C

[0327] The growth inhibition ability of the compounds of the present application on cells expressing KRAS-G12C was evaluated by measuring the cell viability and calculating the GI50 value.

[0328] The tumor cell line NCI-H358 (ATCC catalog number CRL-5807) expressing KRAS-G12C was cultured using RPMI medium added with 10% fetal bovine serum and penicillin / streptomycin double antibiotic; the tumor cell line MIA PaCa2 (ATCC CRL-1420) expressing KRAS-G12C was cultured using DMEM medium added with 10% fetal bovine serum, 2.5% horse serum and penicillin / streptomycin double antibiotic.

[0329] The cells NCI-H358, MIA-Paca2 were seeded in a black clear-bottom 384-well plate (PerkinElmer catalog number 6007460) at a cell density of 1000, 800 cells respectively, and allowed to adhere overnight (8-12 hours). After the cells adhered, the experimental group was added with the compounds of the present application diluted 5 times the working solution concentration (final concentration containing 0.1% dimethyl sulfoxide, i.e. DMSO); the control group was added with the same diluent as the experimental group (final concentration containing 0.1% DMSO). After 72 hours, the amount of cell proliferation was determined by detecting the ATP content using the Cell Titer Glo reagent (Promega catalog number G7572) according to the instructions. The brief operation steps were as follows: the cell plate was taken out and placed at room temperature for 30 minutes; the same volume of Cell Titer Glo reagent as the culture was added; the culture plate was placed on a shaker to lyse for 2 minutes; the culture plate was placed at room temperature for 10 minutes; the light signal value was read using the enzyme-labeled instrument EnVision (PerkinElmer).

[0330] The data of all experimental groups were calculated for the respective inhibition percentage using the DMSO group, and the inhibition rate generated by 9 compound doses diluted by 1 / 3 times was calculated for the GI50 using the data processing software GraphPad. The experimental results are shown in Table 25.

[0331] Table 25

[0332] Compound Number GI 50 (NCI-H358, μM)]]> GI 50 (MIA-Paca2, μΜ)]]> ARS-1620 0.51 1.21 Compound of Formula (I) 0.007 0.004

[0333] Experimental Example 3 Pharmacokinetic Experiment

[0334] This experimental example was conducted to evaluate the pharmacokinetics of the compound in mice by intravenous injection and oral administration.

[0335] Experimental method and condition: Male ICR mice were administered with a single dose of 1 mg / Kg (intravenous injection, solvent 5% DMSO + 15% Solutol + 80% saline) and 5 mg / Kg (oral administration, solvent 1% Tween80 / 2% HPMC / 97% water) respectively. Blood samples were collected from the orbital vein at 5, 15, 30 min, 1, 2, 4, 6, 8, 24 hr after administration, with each sample of about 0.20 mL, and anticoagulated with sodium heparin. The samples were placed on ice after collection, and centrifuged to separate the plasma within 1 hour. The plasma concentration was detected by liquid chromatography tandem mass spectrometry (LC / MS / MS), and the detected concentration was used to calculate the pharmacokinetic parameters. The results are shown in Tables 26 and 27.

[0336] Table 26 Pharmacokinetics of intravenous administration (1 mg / kg)

[0337] Compound T 1 / 2 (hr) AUC inf (ng*hr / mL)]]> Vz (mL / Kg) Cl (mL / min / kg) AMG 510 0.26 176.19 2159.04 94.59 Compound of Formula (I) 1.43 368.33 5615.42 45.25

[0338] Table 27 Pharmacokinetics of oral administration (5 mg / kg)

[0339] Compound T 1 / 2 (hr) C max (ng / mL) AUC inf (ng*hr / mL)]]> F(％) AMG 510 0.57 177.00 155.14 17.61 Compound of Formula (I) 1.06 108.72 421.20 22.87

[0340] Conclusion: It can be seen that the compound of the present application has good pharmacokinetic absorption in mice, and has pharmacokinetic advantages.

[0341] Experimental Example 4 Xenotransplantation Experiment

[0342] Nu / Nu Nude female mice (n = 7-10) were housed in groups of five animals per cage and allowed free access to tap water and commercial rodent chow (Harlan Teklad 22 / 5 Rodent Diet-8640). Cell line xenograft experiments were performed to allow NCI-H358 tumors to grow in mice. Once the tumor size reached 300 mm 3Animals were randomized and treated with vehicle control (1% Tween 80 + 1% HPMC) or compounds (doses: 10 mg / kg / day, 30 mg / kg / day, 100 mg / kg / day, p.o., respectively). Tumor volume was calculated using the formula) 0.5 X length X width X width. At the end of the experiment, animals were sacrificed, tumors were collected, weighed, and stored for additional analysis.

[0343] Results of the tumor volume change are shown in Table 2. Figure 10 Results of the tumor volume change are shown in Table 2. Figure 11 Results of the tumor volume change are shown in Table 2.

Claims

1. The sulfate of the compound shown in formula (Ⅰ), The sulfate contains 0.5 to 3.0 molar equivalents of sulfuric acid relative to compound (I).

2. The crystal form A of the sulfate of the compound shown in formula (Ⅰ), characterized in that, The X-ray powder diffraction pattern of crystal form A has characteristic diffraction peaks at the following 2θ angles: 7.17±0.2°, 11.93±0.2°, 12.34±0.2°, 12.99±0.2°, 16.80±0.2°, and 17.75±0.2°. The compound represented by formula (Ⅰ) is shown below:

3. The crystal form A according to claim 2, characterized in that, The X-ray powder diffraction pattern of crystal form A has characteristic diffraction peaks at the following 2θ angles: 7.17±0.2°, 11.93±0.2°, 12.34±0.2°, 12.99±0.2°, 13.54±0.2°, 16.80±0.2°, 17.75±0.2°, 19.09±0.2°, 20.91±0.2°, and 21.71±0.2°.

4. The crystal form A according to claim 3, characterized in that, The X-ray powder diffraction pattern of crystal form A is substantially as shown in Figure 1.

5. Crystal form B of the sulfate of the compound shown in formula (Ⅰ), characterized in that, The X-ray powder diffraction pattern of crystal form B has characteristic diffraction peaks at the following 2θ angles: 5.67±0.2°, 8.78±0.2°, 10.28±0.2°, 11.02±0.2°, 12.19±0.2°, and 15.51±0.2°. The compound represented by formula (Ⅰ) is shown below:

6. The crystal form B according to claim 5, characterized in that, The X-ray powder diffraction pattern of crystal form B has characteristic diffraction peaks at the following 2θ angles: 5.67±0.2°, 8.78±0.2°, 10.28±0.2°, 11.02±0.2°, 12.19±0.2°, 12.60±0.2°, 13.14±0.2°, 15.51±0.2°, 15.99±0.2°, and 19.56±0.2°.

7. The crystal form B according to claim 5, characterized in that, The X-ray powder diffraction pattern of crystal form B is substantially as shown in Figure 2.

8. The crystal form C of the sulfate of the compound shown in formula (I), characterized in that, The X-ray powder diffraction pattern of crystal form C has characteristic diffraction peaks at the following 2θ angles: 5.51±0.2°, 7.71±0.2°, 11.90±0.2°, 13.67±0.2°, 15.69±0.2°, and 20.15±0.2°. The compound represented by formula (Ⅰ) is shown below:

9. The crystal form C according to claim 8, characterized in that, The X-ray powder diffraction pattern of crystal form C is essentially the X-ray powder diffraction pattern shown in Figure 3.

10. The crystal form D of the sulfate of the compound shown in formula (I), characterized in that, The X-ray powder diffraction pattern of the crystal form D has characteristic diffraction peaks at the following 2θ angles: 6.85±0.2°, 10.29±0.2°, 12.78±0.2°, 16.36±0.2°, 19.84±0.2°, and 20.66±0.2°. The compound represented by formula (Ⅰ) is shown below:

11. The crystal form D according to claim 10, characterized in that, The X-ray powder diffraction pattern of the crystal form D has essentially the X-ray powder diffraction pattern shown in Figure 4.

12. The crystal form E of the sulfate of the compound shown in formula (I), characterized in that, The X-ray powder diffraction pattern of crystal form E has characteristic diffraction peaks at the following 2θ angles: 5.52±0.2°, 8.93±0.2°, 11.04±0.2°, 12.16±0.2°, 12.82±0.2°, and 22.67±0.2°. The compound represented by formula (Ⅰ) is shown below:

13. The crystal form E according to claim 12, characterized in that, The X-ray powder diffraction pattern of crystal form E has characteristic diffraction peaks at the following 2θ angles: 5.52±0.2°, 8.52±0.2°, 8.93±0.2°, 11.04±0.2°, 12.16±0.2°, 12.82±0.2°, 14.63±0.2°, 15.34±0.2°, 16.88±0.2°, and 22.67±0.2°.

14. The crystal form E according to claim 12, characterized in that, The X-ray powder diffraction pattern of the crystal form E is essentially the X-ray powder diffraction pattern shown in Figure 5.

15. The crystal form F of the sulfate of the compound shown in formula (I), characterized in that, The X-ray powder diffraction pattern of the crystal form F has characteristic diffraction peaks at the following 2θ angles: 5.40±0.2°, 13.01±0.2°, 18.09±0.2°, and 21.65±0.2°. The compound represented by formula (Ⅰ) is shown below:

16. The crystal form F according to claim 15, characterized in that, The X-ray powder diffraction pattern of the crystal form F is essentially the X-ray powder diffraction pattern shown in Figure 6.

17. The crystal form G of the sulfate of the compound shown in formula (I), characterized in that, The X-ray powder diffraction pattern of the crystal form G has characteristic diffraction peaks at the following 2θ angles: 5.88±0.2°, 7.05±0.2°, 9.39±0.2°, 10.87±0.2°, 12.00±0.2°, and 16.15±0.2°. The compound represented by formula (Ⅰ) is shown below:

18. The crystal form G according to claim 17, characterized in that, The X-ray powder diffraction pattern of the crystal form G is substantially as shown in Figure 7.

19. The sulfate of the compound shown in formula (I) has a crystal form H, characterized in that, The X-ray powder diffraction pattern of the crystal form H has characteristic diffraction peaks at the following 2θ angles: 6.11±0.2°, 11.59±0.2°, 12.19±0.2°, 15.66±0.2°, 18.34±0.2°, and 13.82±0.2°. The compound represented by formula (Ⅰ) is shown below:

20. The crystal form H according to claim 19, characterized in that, The X-ray powder diffraction pattern of the crystal form H has essentially the X-ray powder diffraction pattern shown in Figure 8.

21. Benzenesulfonates of the compounds shown in formula (I), which are shown below: The benzenesulfonate contains benzenesulfonic acid in a molar equivalent of 0.5-3.0 molar equivalents relative to compound (I).

22. The benzenesulfonate of the compound shown in formula (Ⅰ) in crystal form I, characterized in that, The X-ray powder diffraction pattern of crystal form I has characteristic diffraction peaks at the following 2θ angles: 7.63±0.2°, 12.05±0.2°, 13.78±0.2°, 15.09±0.2°, 16.04±0.2°, and 18.35±0.2°. The compound represented by formula (Ⅰ) is shown below:

23. The crystal form I according to claim 22, characterized in that, The X-ray powder diffraction pattern of crystal form I has characteristic diffraction peaks at the following 2θ angles: 7.63±0.2°, 11.35±0.2°, 12.05±0.2°, 13.78±0.2°, 15.09±0.2°, 16.04±0.2°, 16.65±0.2°, 18.35±0.2°, 18.87±0.2°, and 23.53±0.2°.

24. The crystal form I according to claim 23, characterized in that, The X-ray powder diffraction pattern of crystal form I is substantially as shown in Figure 9.

25. A pharmaceutical composition, characterized in that, The pharmaceutical composition contains a sulfate of the compound of formula (I) as claimed in claim 1, or crystal form A as claimed in any one of claims 2-4, or crystal form B as claimed in any one of claims 5-7, or crystal form C as claimed in any one of claims 8-9, or crystal form D as claimed in any one of claims 10-11, or crystal form E as claimed in any one of claims 12-14, or crystal form F as claimed in any one of claims 15-16, or crystal form G as claimed in any one of claims 17-18, or crystal form H as claimed in any one of claims 19-20, or a benzenesulfonate of the compound of formula (I) as claimed in claim 21, or crystal form I as claimed in any one of claims 22-24.

26. Use of the sulfate of the compound of formula (I) as claimed in claim 1, or crystal form A as claimed in any one of claims 2-4, or crystal form B as claimed in any one of claims 5-7, or crystal form C as claimed in any one of claims 8-9, or crystal form D as claimed in any one of claims 10-11, or crystal form E as claimed in any one of claims 12-14, or crystal form F as claimed in any one of claims 15-16, or crystal form G as claimed in any one of claims 17-18, or crystal form H as claimed in claims 19-20, or benzenesulfonate of the compound of formula (I) as claimed in claim 21, or crystal form I as claimed in any one of claims 22-24, or the pharmaceutical composition as claimed in claim 25, in the preparation of a medicament for the prevention and / or treatment of KRAS-G12C-related diseases.

27. The use according to claim 26, characterized in that, The KRAS-G12C-related diseases mentioned are selected from non-small cell lung cancer, colon cancer, and pancreatic cancer.

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