WO2022048545A1 - Crystal form of pyridopyrimidine compound - Google Patents

Abstract

Disclosed are a crystal form of a pyridopyrimidine compound and a preparation method therefor, and further comprised is the use of the crystal form in the preparation of a drug for treating cancer.

Description

A crystal form of a pyridopyrimidine compound

This application claims the following priority:

PCT/CN2020/112863, filed on September 1, 2020.

technical field

The invention relates to a crystal form of a pyridopyrimidine compound and a preparation method thereof, and also includes the application of the crystal form in the preparation of a drug for treating cancer.

Background technique

The first RAS oncogene was found in rat sarcoma, hence the name. RAS protein is the product expressed by the RAS gene, which refers to a class of closely related monomeric globulins composed of 189 amino acids with a molecular weight of 21KDa. It can bind to guanine trinucleotide phosphate (GTP) or guanine dinucleotide phosphate (GDP). The active state of RAS protein has effects on cell growth, differentiation, cytoskeleton, protein transport and secretion, and its activity is regulated by binding to GTP or GDP. When the RAS protein binds to GDP, it is in a dormant state, that is, in an "inactive" state; when stimulated by upstream specific cell growth factors, the RAS protein is induced to exchange GDP and bind to GTP, which is called "activation". condition. The RAS protein bound to GTP can activate downstream proteins for signal transmission. The RAS protein itself has weak hydrolysis GTP hydrolysis activity and can hydrolyze GTP to GDP. In this way, the transition from the activated state to the deactivated state can be achieved. In this hydrolysis process, GAP (GTPase activating proteins, GTP hydrolase activating proteins) is also required. It can interact with RAS protein, greatly promoting its ability to hydrolyze GTP to GDP. Mutation of the RAS protein will affect its interaction with GAP, which also affects its ability to hydrolyze GTP to GDP, making it always active. Activated RAS proteins continue to give downstream proteins growth signals, which eventually lead to the continuous growth and differentiation of cells, and ultimately produce tumors. There are many members of the RAS gene family, among which subfamilies closely related to various cancers mainly include Kirsten rat sarcoma virus oncogene homolog (KRAS), Harvey rat sarcoma virus oncogene homolog (HRAS) and neuronal Blastoma rat sarcoma virus oncogene homolog (NRAS). It has been found that about 30% of human tumors carry some mutated RAS gene, among which KRAS mutation is the most significant, accounting for 86% of all RAS mutations. For KRAS mutations, the most common mutations occurred at residues 12 glycine (G12), 13 glycine (G13) and 61 glutamine (Q61), of which G12 mutations accounted for 83%.

The G12C mutation is one of the more common mutations in the KRAS gene, which refers to the mutation of glycine No. 12 to cysteine. KRAS G12C mutation is the most common in lung cancer. According to the data reported in the literature (Nat Rev Drug Discov 2014; 13:828-851), KRAS G12C mutation accounts for about 10% of all lung cancer patients.

As a cutting-edge target, the KRAS G12C mutant protein has not been studied much. Literature (Nature. 2013; 503: 548-551) reported a class of covalently bound inhibitors targeting KRAS G12C mutation, but these compounds have low enzymatic activity and no significant activity at the cellular level. Literature (Science 2016; 351: 604-608, Cancer Discov 2016; 6: 316-29) reported that a class of compounds exhibited μM-level anti-proliferative activity at the cellular level, but its structural and metabolic stability was poor and the activity was also very high. Difficult to further improve. In addition to a few literature reports, Araxes Pharma LLC has applied for several patents for KRAS G12C inhibitors, such as WO2016164675 and WO2016168540, which reported a class of quinazoline derivatives with high enzyme-binding activity and exhibited μM level The cell anti-proliferative activity, its structure is stable, and has a certain selectivity. These compounds all have an acrylamide fragment, which acts as a Michael addition receptor and forms a covalently bound complex with the G12C residue on the KRAS protein. In 2018, LiuYi et al. published in Cell (Matthew R.Janes, Yi Liu et al., Cell, 2018, 172, 578–589.) a covalently bound inhibitor ARS-1620 targeting KRAS G12C mutation, the compound It has good metabolic stability, exhibits nM-level cell anti-proliferation activity at the cellular level, and can effectively inhibit tumor growth in the pancreatic cancer MIA-Paca2 cell subcutaneous xenograft tumor model. In the second half of 2018, Amgen's KRAS G12C inhibitor AMG 510 started Phase I clinical recruitment (NCT03600883), which is the first organic small molecule KRAS G12C inhibitor to enter clinical research. At the 2019 AACR meeting, AMG 510's The structural formula and some preclinical research data were published. At the ASCO meeting in 2019, the early phase I clinical results of AMG 510 were announced. The control rate reaches 90%. In addition, the KRAS G12C inhibitor MRTX849 developed by Mirati Therapeutics also started Phase I clinical recruitment in January 2019 (NCT03785249), and its representative patents are WO2017201161 and WO2019099524.

SUMMARY OF THE INVENTION

The present invention provides a crystal form of compound A of formula (I), the X-ray powder diffraction pattern of which has characteristic diffraction peaks at the following 2θ angles: 9.96±0.20°, 19.83±0.20°, 22.23±0.20°.

In some embodiments of the present invention, the above-mentioned crystal form A, its X-ray powder diffraction pattern has characteristic diffraction peaks at the following 2θ angles: 9.96±0.20°, 18.09±0.20°, 19.83±0.20°, 22.23±0.20°, 22.65 ±0.20°, 24.80±0.20°, 25.78±0.20°, 27.42±0.20°.

In some embodiments of the present invention, the above-mentioned A crystal form, its X-ray powder diffraction pattern has characteristic diffraction peaks at the following 2θ angles: 5.921°, 9.960°, 11.718°, 15.071°, 15.584°, 17.401°, 18.089°, 18.954°, 19.826°, 22.231°, 22.646°, 23.377°, 24.796°, 25.783°, 27.417°, 29.860°, 30.334°, 30.649°, 32.095°, 33.691°, 35.187°, 38.757°, 39.073°.

The present invention provides a crystal form of compound A of formula (I), whose X-ray powder diffraction pattern has characteristic diffraction peaks at the following 2θ angles: 9.96±0.20°, 19.83±0.20°, and also 22.23±0.20°, and/or 5.92± 0.20°, and/or 11.72±0.20°, and/or 15.07±0.20°, and/or 15.58±0.20°, and/or 17.40±0.20°, and/or 18.09±0.20°, and/or 18.95±0.20° , and/or 22.65±0.20°, and/or 23.38±0.20°, and/or 24.80±0.20°, and/or 25.78±0.20°, and/or 27.42±0.20°, and/or 29.86±0.20°, and /or 30.33±0.20°, and/or 30.65±0.20°, and/or 32.10±0.20°, and/or 33.69±0.20°, and/or 35.19±0.20°, and/or 38.76±0.20°, and/or There are characteristic diffraction peaks at 39.07±0.20°.

In some embodiments of the present invention, the XRPD pattern of the above-mentioned crystal form A is shown in FIG. 1 .

In some schemes of the present invention, the XRPD spectrum analysis data of above-mentioned A crystal form is shown in Table 1:

Table 1 XRPD pattern analysis data of compound A crystal form of formula (I)

serial number	2θ angle (°)	Surface spacing (A)	Relative Strength(%)	serial number	2θ angle (°)	Surface spacing (A)	Relative Strength(%)
1	5.921	14.9145	8.3	13	24.796	3.5877	40.9
2	9.960	8.8738	47.1	14	25.783	3.4526	25.8
3	11.718	7.5459	21.2	15	27.417	3.2503	31.8
4	15.071	5.8735	8.2	16	29.860	2.9897	8
5	15.584	5.6816	24.7	17	30.334	2.9441	18.9
6	17.401	5.0921	22.5	18	30.649	2.9146	12.5
7	18.089	4.8998	25.1	19	32.095	2.7865	8.7
8	18.954	4.6783	15.5	20	33.691	2.6581	8.8
9	19.826	4.4744	45.5	twenty one	35.187	2.5484	7
10	22.231	3.9956	100	twenty two	38.757	2.3215	8.6
11	22.646	3.9231	41.3	twenty three	39.073	2.3034	6.7
12	23.377	3.8021	twenty three	/	/	/	/

In some embodiments of the present invention, the above-mentioned crystal form A, the differential scanning calorimetry curve of which has an onset of an endothermic peak at 249.10±5°C.

In some embodiments of the present invention, the above-mentioned crystal form A, the differential scanning calorimetry curve of which has an onset of an endothermic peak at 249.10±3°C.

In some embodiments of the present invention, the DSC spectrum of the above-mentioned crystal form A is shown in FIG. 2 .

In some embodiments of the present invention, the above-mentioned crystal form A has a weight loss of 0.05409% in the thermogravimetric analysis curve at 120.17±3°C.

In some embodiments of the present invention, the above-mentioned crystal form A, its TGA spectrum is shown in FIG. 3 .

The present invention also provides compounds of formula (II)

The present invention also provides compound B crystal form of formula (II), whose X-ray powder diffraction pattern has characteristic diffraction peaks at the following 2θ angles: 10.16±0.20°, 17.75±0.20°, 24.32±0.20°.

In some embodiments of the present invention, the above-mentioned crystal form B, its X-ray powder diffraction pattern has characteristic diffraction peaks at the following 2θ angles: 10.16±0.20°, 17.75±0.20°, 19.25±0.20°, 19.98±0.20°, 22.59 ±0.20°, 23.06±0.20°, 24.32±0.20°, 30.71±0.20°.

In some embodiments of the present invention, the above-mentioned crystal form B, its X-ray powder diffraction pattern has characteristic diffraction peaks at the following 2θ angles: 4.105°, 8.800°, 10.164°, 11.633°, 13.060°, 14.973°, 15.324°, 16.274°, 16.613°, 17.102°, 17.752°, 18.107°, 19.034°, 19.254°, 19.983°, 20.912°, 22.586°, 23.060°, 24.322°, 24.698°, 25.681°, 26.157°, 27.216° , 28.262°, 28.627°, 29.353°, 30.277°, 30.711°, 32.386°, 33.335°, 34.139°, 35.010°, 35.448°, 35.859°, 36.608°, 37.976°.

The present invention provides compound B crystal form of formula (I), the X-ray powder diffraction pattern of which has characteristic diffraction peaks at the following 2θ angles: 10.16±0.20°, 17.75±0.20°, and also 24.32±0.20°, and/or 4.11± 0.20°, and/or 8.80±0.20°, and/or 11.63±0.20°, and/or 13.06±0.20°, and/or 14.97±0.20°, and/or 15.32±0.20°, and/or 16.27±0.20° , and/or 16.61±0.20°, and/or 17.10±0.20°, and/or 18.11±0.20°, and/or 19.03±0.20°, and/or 19.25±0.20°, and/or 19.98±0.20°, and /or 20.91±0.20°, and/or 22.59±0.20°, and/or 23.06±0.20°, and/or 24.70±0.20°, and/or 25.68±0.20°, and/or 26.16±0.20°, and/or 27.22±0.20°, and/or 27.67±0.20°, and/or 28.26±0.20°, and/or 28.63±0.20°, and/or 29.35±0.20°, and/or 30.28±0.20°, and/or 30.71± 0.20°, and/or 32.39±0.20°, and/or 33.34±0.20°, and/or 34.14±0.20°, and/or 35.01±0.20°, and/or 35.45±0.20°, and/or 35.86±0.20° , and/or 36.61±0.20°, and/or 37.98±0.20° have characteristic diffraction peaks.

In some embodiments of the present invention, the XRPD pattern of the above-mentioned crystal form B is shown in FIG. 4 .

In some schemes of the present invention, the XRPD spectrum analysis data of above-mentioned B crystal form is shown in Table 2:

Table 2 XRPD pattern analysis data of compound B crystal form of formula (II)

serial number	2θ angle (°)	Surface spacing (A)	Relative Strength(%)	serial number	2θ angle (°)	Surface spacing (A)	Relative Strength(%)
1	4.105	21.5094	1.2	20	24.698	3.6017	53.9
2	8.800	10.0397	13.4	twenty one	25.681	3.4660	7.1
3	10.164	8.6960	75	twenty two	26.157	3.4040	14.8
4	11.633	7.6006	5.7	twenty three	27.216	3.2739	2.2

5	13.060	6.7730	1.8	twenty four	27.674	3.2208	4.6
6	14.973	5.9119	11.2	25	28.262	3.1550	3.6
7	15.324	5.7774	16.2	26	28.627	3.1157	9
8	16.274	5.4420	12.6	27	29.353	3.0402	1.9
9	16.613	5.3317	11.4	28	30.277	2.9496	7.5
10	17.102	5.1803	11.1	29	30.711	2.9088	26.2
11	17.752	4.9922	61.1	30	32.386	2.7621	12.8
12	18.107	4.8950	17.2	31	33.335	2.6856	13.8
13	19.034	4.6588	3.1	32	34.139	2.6242	1.3
14	19.254	4.6061	29.6	33	35.010	2.5609	7.9
15	19.983	4.4396	25.9	34	35.448	2.5302	16.8
16	20.912	4.2445	3.9	35	35.859	2.5021	10.2
17	22.586	3.9335	17.2	36	36.608	2.4527	3.7
18	23.060	3.8537	56.4	37	37.976	2.3674	7
19	24.322	3.6565	100	/	/	/	/

In some embodiments of the present invention, the above-mentioned crystal form B, its differential scanning calorimetry curve has an onset of an endothermic peak at 253.00±5°C.

In some embodiments of the present invention, the above-mentioned crystal form B, its differential scanning calorimetry curve has an onset of an endothermic peak at 253.00±3°C.

In some embodiments of the present invention, the DSC spectrum of the above-mentioned crystal form B is shown in FIG. 5 .

In some embodiments of the present invention, the above-mentioned crystal form B, its thermogravimetric analysis curve has a weight loss of 0.4116% at 150.14±3°C.

In some embodiments of the present invention, the above-mentioned crystal form B, its TGA spectrum is shown in FIG. 6 .

The present invention provides the compound of formula (I) p-toluenesulfonate salt form C, and its X-ray powder diffraction pattern has characteristic diffraction peaks at the following 2θ angles: 19.57±0.20°, 21.56±0.20°, 24.01±0.20°.

In some embodiments of the present invention, the above crystal form C, its X-ray powder diffraction pattern has characteristic diffraction peaks at the following 2θ angles: 12.17±0.20°, 15.78±0.20°, 16.69±0.20°, 19.57±0.20°, 21.56 ±0.20°, 24.01±0.20°, 25.02±0.20°, 34.80±0.20°.

In some embodiments of the present invention, the above-mentioned crystal form C, its X-ray powder diffraction pattern has characteristic diffraction peaks at the following 2θ angles: 8.030°, 12.169°, 13.194°, 14.399°, 15.332°, 15.779°, 16.055°, 16.496°, 16.687°, 18.996°, 19.289°, 19.567°, 20.556°, 20.987°, 21.562°, 21.897°, 22.864°, 24.006°, 24.281°, 25.018°, 26.433°, 28.397°, 28.82° , 31.066°, 31.773°, 31.935°, 34.795°.

The present invention provides compound C of formula (I), the X-ray powder diffraction pattern of which has characteristic diffraction peaks at the following 2θ angles: 19.57±0.20°, 21.56±0.20°, and also 24.01±0.20°, and/or 8.03±0.20° 0.20°, and/or 12.17±0.20°, and/or 13.19±0.20°, and/or 14.40±0.20°, and/or 15.33±0.20°, and/or 15.78±0.20°, and/or 16.06±0.20° , and/or 16.50±0.20°, and/or 16.69±0.20°, and/or 19.00±0.20°, and/or 19.29±0.20°, and/or 20.56±0.20°, and/or 20.99±0.20°, and /or 21.90±0.20°, and/or 22.86±0.20°, and/or 24.28±0.20°, and/or 25.02±0.20°, and/or 26.43±0.20°, and/or 28.40±0.20°, and/or 28.80±0.20°, and/or 30.43±0.20°, and/or 31.07±0.20°, and/or 31.77±0.20°, and/or 31.94±0.20°, and/or 34.80±0.20°, and/or 38.48± There is a characteristic diffraction peak at 0.20°.

In some embodiments of the present invention, the X-ray powder diffraction pattern of the above crystal form C is shown in FIG. 7 .

In some schemes of the present invention, the XRPD spectrum analysis data of above-mentioned C crystal form is shown in Table 3:

Table 3 XRPD spectrum analysis data of compound p-toluenesulfonate salt form C of formula (I)

serial number	2θ angle (°)	Surface spacing (A)	Relative Strength(%)	serial number	2θ angle (°)	Surface spacing (A)	Relative Strength(%)
1	8.030	11.001	12.7	16	21.897	4.0557	12.8
2	12.169	7.2671	17.9	17	22.864	3.8863	7.7
3	13.194	6.7049	9.1	18	24.006	3.7040	100
4	14.399	6.1462	13.3	19	24.281	3.6627	87
5	15.332	5.7744	10.6	20	25.018	3.5564	23.1
6	15.779	5.6118	14.8	twenty one	26.433	3.3691	7.4
7	16.055	5.5158	14.9	twenty two	28.397	3.1404	11
8	16.496	5.3694	7.5	twenty three	28.802	3.0972	9.6
9	16.687	5.3084	13.9	twenty four	30.430	2.9351	7.5
10	18.996	4.6680	6	25	31.066	2.8764	13.8
11	19.289	4.5976	17.7	26	31.773	2.8140	8.3
12	19.567	4.5330	33.8	27	31.935	2.8001	8.3
13	20.556	4.3170	12.9	28	34.795	2.5762	17.5
14	20.987	4.2294	7.5	29	38.484	2.3373	5.4
15	21.562	4.1180	48.1	/	/	/	/

In some embodiments of the present invention, the above-mentioned crystal form C, the differential scanning calorimetry curve of which has an onset of an endothermic peak at 262.38±5°C.

In some embodiments of the present invention, the above-mentioned crystal form C, the differential scanning calorimetry curve of which has an onset of an endothermic peak at 262.38±3°C.

In some embodiments of the present invention, the DSC spectrum of the above-mentioned crystal form C is shown in FIG. 8 .

In some embodiments of the present invention, the above-mentioned crystal form C, its thermogravimetric analysis curve has a weight loss of 1.542% at 146.27±3°C.

In some embodiments of the present invention, the above-mentioned crystal form C, its TGA spectrum is shown in FIG. 9 .

The present invention provides the compound of formula (I) hydrochloride salt form D, and its X-ray powder diffraction pattern has characteristic diffraction peaks at the following 2θ angles: 17.50±0.20°, 24.50±0.20°, 25.21±0.20°.

In some embodiments of the present invention, the above-mentioned crystal form D, its X-ray powder diffraction pattern has characteristic diffraction peaks at the following 2θ angles: 16.39±0.20°, 17.50±0.20°, 19.49±0.20°, 19.96±0.20°, 22.09 ±0.20°, 24.50±0.20°, 25.21±0.20°, 27.85±0.20°.

In some embodiments of the present invention, the above-mentioned crystal form D, its X-ray powder diffraction pattern has characteristic diffraction peaks at the following 2θ angles: 6.742°, 10.435°, 11.853°, 16.392°, 16.927°, 17.499°, 18.248°, 19.487°, 19.958°, 20.317°, 22.091°, 23.558°, 24.498°, 25.207°, 26.215°, 27.851°, 28.506°, 29.667°, 34.121°, 37.360°.

The present invention provides a crystalline form of compound D of formula (I), whose X-ray powder diffraction pattern has characteristic diffraction peaks at the following 2θ angles: 17.50±0.20°, 24.50±0.20°, and also 25.21±0.20°, and/or 6.74± 0.20°, and/or 10.44±0.20°, and/or 11.85±0.20°, and/or 16.39±0.20°, and/or 16.93±0.20°, and/or 18.25±0.20°, and/or 19.49±0.20° , and/or 19.96±0.20°, and/or 20.32±0.20°, and/or 22.09±0.20°, and/or 23.56±0.20°, and/or 26.22±0.20°, and/or 27.85±0.20°, and Characteristic diffraction peaks at 28.51±0.20°, and/or 29.67±0.20°, and/or 34.12±0.20°, and/or 37.36±0.20°.

In some embodiments of the present invention, the X-ray powder diffraction pattern of the above-mentioned D crystal form is shown in FIG. 10 .

In some schemes of the present invention, the XRPD spectrum analysis data of above-mentioned D crystal form is shown in Table 4:

Table 4 XRPD pattern analysis data of compound hydrochloride D crystal form of formula (I)

serial number	2θ angle (°)	Surface spacing (A)	Relative Strength(%)	serial number	2θ angle (°)	Surface spacing (A)	Relative Strength(%)
1	6.742	13.0998	5	11	22.091	4.0205	38.9
2	10.435	8.4704	28.2	12	23.558	3.7733	7.2
3	11.853	7.4599	15.5	13	24.498	3.6307	100
4	16.392	5.4031	34.7	14	25.207	3.5301	75.2
5	16.927	5.2335	15.4	15	26.215	3.3966	4.8
6	17.499	5.0639	72.9	16	27.851	3.2007	31.6
7	18.248	4.8575	5.5	17	28.506	3.1286	3.4
8	19.487	4.5514	35.9	18	29.667	3.0088	18.1
9	19.958	4.445	57.9	19	34.121	2.6255	12.1
10	20.317	4.3674	28.6	20	37.360	2.405	13.9

In some embodiments of the present invention, the above-mentioned crystal form D, the differential scanning calorimetry curve of which has an onset of an endothermic peak at 171.38±5°C.

In some embodiments of the present invention, the above-mentioned crystal form D, its differential scanning calorimetry curve has an onset of an endothermic peak at 171.38±3°C.

In some embodiments of the present invention, the DSC spectrum of the above-mentioned D crystal form is shown in FIG. 11 .

In some embodiments of the present invention, the above-mentioned crystal form D, its thermogravimetric analysis curve has a weight loss of 0.7461% at 99.76±3°C, and a weight loss of 5.3141% at 169.75±3°C.

In some embodiments of the present invention, the TGA pattern of the above-mentioned D crystal form is shown in FIG. 12 .

The present invention also provides the above-mentioned formula (I) compound A crystal form, formula (II) compound B crystal form, formula (I) compound p-toluenesulfonate salt crystal form C and formula (I) compound hydrochloride salt crystal form D in preparation Application in the treatment of cancer drugs.

In some embodiments of the present invention, the aforementioned cancers include lung cancer, lymphoma, esophageal cancer, ovarian cancer, pancreatic cancer, rectal cancer, glioma, cervical cancer, urothelial cancer, gastric cancer, endometrial cancer, liver cancer , cholangiocarcinoma, breast cancer, colon cancer, leukemia and melanoma.

technical effect

The compound of the present invention has good PK property and oral absorption rate, its crystal form is relatively stable, its solubility is good, its hygroscopicity is appropriate, and it is less affected by light and heat, and has good druggability.

Definition and Explanation

Unless otherwise specified, the following terms and phrases used herein are intended to have the following meanings. A particular phrase or term should not be considered indeterminate or unclear without a specific definition, but should be understood in its ordinary meaning. When a trade name appears herein, it is intended to refer to its corresponding commercial product or its active ingredient.

The structure of the compound of the present invention can be confirmed by conventional methods well known to those skilled in the art. If the present invention relates to the absolute configuration of the compound, the absolute configuration can be confirmed by conventional technical means in the art.

The intermediate compounds of the present invention can be prepared by a variety of synthetic methods well known to those skilled in the art, including the specific embodiments listed below, the embodiments formed by their combination with other chemical synthesis methods, and those skilled in the art. Well-known equivalents, preferred embodiments include, but are not limited to, the examples of the present invention.

The chemical reactions of specific embodiments of the present invention are carried out in suitable solvents suitable for the chemical changes of the present invention and their required reagents and materials. In order to obtain the compounds of the present invention, it is sometimes necessary for those skilled in the art to modify or select the synthetic steps or reaction schemes on the basis of the existing embodiments.

The present invention will be specifically described below through examples, which do not imply any limitation to the present invention.

All solvents used in the present invention are commercially available and used without further purification.

The following abbreviations are used in the present invention: rt stands for room temperature; THF stands for tetrahydrofuran; CDI stands for carbonyldiimidazole; DCM stands for dichloromethane; DMF stands for N,N-dimethylformamide; MeOH stands for methanol; MsOH stands for methanesulfonic acid ; acetone stands for acetone; NIS stands for N-iodosuccinimide; HPLC stands for high performance liquid chromatography; TLC stands for thin layer chromatography.

The structure of the compound of the present invention can be confirmed by conventional methods well known to those skilled in the art. If the present invention relates to the absolute configuration of the compound, the absolute configuration can be confirmed by conventional technical means in the art. For example, single crystal X-ray diffractometry (SXRD), the cultivated single crystal is collected by Bruker D8venture diffractometer, the light source is CuKα radiation, and the scanning method is as follows:

After scanning and collecting relevant data, the crystal structure was further analyzed by the direct method (Shelxs97), and the absolute configuration could be confirmed.

Compounds are named according to conventional nomenclature in the art or

Software naming, commercially available compounds use supplier catalog names.

1. Instruments and analytical methods

1.1. Powder X-ray diffraction (X-ray powder diffractometer, XRPD)

Instrument model: Bruker D8advance X-ray diffractometer

Test conditions: The detailed XRPD parameters are as follows:

X-ray generator: Cu, kα,

Tube voltage: 40kV, tube current: 40mA.

Scattering slit: 0.60mm

Detector slit: 10.50mm

Anti-scatter slit: 7.10mm

Scanning range: 3-40 degrees

Step: 0.02 degrees

Step size: 0.12 seconds

Sample tray speed: 15rpm

1.2. Differential Thermal Analysis (Differential Scanning Calorimeter, DSC)

Instrument model: TA Q2000 Differential Scanning Calorimeter

Test conditions: Take a sample (0.5-1 mg) and place it in a DSC aluminum pot for testing.

1.3. Thermal Gravimetric Analyzer (TGA)

Instrument Model: TA Q5000IR Thermogravimetric Analyzer

Test conditions: take a sample (2-5 mg) and place it in a TGA platinum pot for testing.

Description of drawings

Fig. 1 is the XRPD spectrum of compound A of formula (I).

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

Figure 3 is a TGA spectrum of the crystal form of compound A of formula (I).

Fig. 4 is the XRPD spectrum of the crystal form of compound B of formula (II).

Figure 5 is the DSC spectrum of the crystalline form of compound B of formula (II).

Figure 6 is a TGA spectrum of the crystalline form of compound B of formula (II).

Figure 7 is the XRPD spectrum of the compound of formula (I) p-toluenesulfonate salt form C.

Figure 8 is the DSC spectrum of the compound of formula (I) p-toluenesulfonate salt form C.

Figure 9 is the TGA spectrum of the compound of formula (I) p-toluenesulfonate salt form C.

Figure 10 is the XRPD spectrum of the compound of formula (I) hydrochloride salt form D.

Fig. 11 is the DSC spectrum of the crystalline form D of the compound of formula (I) hydrochloride.

Figure 12 is the TGA spectrum of the compound of formula (I) hydrochloride salt form D.

Figure 13 is an ellipsoid diagram of the three-dimensional structure of the compound of formula (I).

detailed description

In order to better understand the content of the present invention, further description will be given below in conjunction with specific embodiments, but the specific embodiments do not limit the content of the present invention.

Example 1: Preparation of compounds of formula (I)

first step:

A solution of compound 1 (1 kg, 6.41 mol, 1 equiv) and potassium carbonate (1.77 kg, 12.81 mol, 2 equiv) in acetonitrile (15 L) was cooled to 0°C, then compound 2 (2.62 kg, 19.22 mol, 2.05 liter, 3 equiv.) was added dropwise to the above solution (about 2 hours), and the mixture was heated to 25°C after the dropwise addition and stirred for 16 hours. Saturated aqueous sodium carbonate solution (5 L) and water (5 L) were successively added to the reaction system to quench the reaction, the aqueous phase was extracted with ethyl acetate (5 L) after separation, and the combined organic phases were washed with saturated brine ( 8 L), and the organic phase was concentrated under reduced pressure. The obtained residue was dispersed in a mixed solvent of petroleum ether:ethyl acetate=3:1 (16 L), and stirred at 25°C for 16 hours. The reaction system was filtered, and the filter cake was washed with petroleum ether:ethyl acetate=3:1 (2 liters), and then placed in a vacuum drying oven (50°C, -0.1MPa) for drying for 16 hours to obtain compound 3. ¹ H NMR (400 MHz, CD ₃ OD) δ 7.84 (td, J=8.2, 1.4 Hz, 1H), 7.62-7.56 (m, 1H), 7.55-7.48 (m, 1H), 3.78 (s, 3H) , 3.66-3.47 (m, 2H).

Step 2:

To a solution of compound 3 (1.42 kg, 5.14 mol, 1 equiv) in acetonitrile (15 L) was added cesium carbonate (1.67 kg, 5.14 mol, 1 equiv) followed by compound 4 (881.08 g, 5.24 mol, 1 hr) 746.68 mL, 1.02 equiv.) was added dropwise to the above mixed solution, and the resulting reaction solution was stirred at 25°C for 2 hours. The reaction mixture was filtered, the filter cake was washed with ethyl acetate (2 L), the filtrate was concentrated under reduced pressure, the obtained residue was diluted with ethyl acetate (2 L), followed by water (8 L) and saturated brine (6 L) ), the organic phase was dried over anhydrous sodium sulfate and filtered, the filter cake was washed with ethyl acetate (2 L), and the filtrate was concentrated under reduced pressure. The resulting crude product was dispersed in methyl tert-butyl ether (10 L) and stirred at 25°C for 16 hours. The reaction mixture was filtered, and the filter cake was dried in a vacuum drying oven (50°C, -0.1 MPa) for 16 hours to obtain compound 5.

third step:

To a solution of compound 5 (2370 g, 6.27 mol, 1 equiv) in trifluoroethanol (12 L) was added triethylamine (1.27 kg, 12.53 mol, 1.74 L, 2 equiv) and the resulting mixture was stirred at 80°C for 16 Hour. HPLC showed that the reaction of the starting material was complete, about 85% of the target product was formed, and about 5% of the intermediate was not completely converted. The above reaction solution was further stirred at 80°C for 8 hours. The reaction solution was concentrated under reduced pressure, the obtained residue was dissolved in ethyl acetate (20 liters) and water (10 liters), the pH of the aqueous phase was adjusted to 2 with 2 mol/liter aqueous hydrochloric acid, and the separated organic phase was used Washed with saturated brine (5L*2), dried over anhydrous sodium sulfate and filtered, the filtrate was concentrated under reduced pressure to obtain a crude product dispersed in a mixed solution of ethanol (2L) and methyl tert-butyl ether (2L) , and stirred at 20°C for 1 hour. The reaction mixture was filtered, the filter cake was washed with ethanol (1 L), the filter cake was dissolved in ethanol (5.5 L), heated to 80°C, then the reaction system was cooled to 15°C and stirred for 14 hours. The reaction mixture was filtered and the filter cake was washed with ethanol (0.5 L) and dried in vacuo to give compound 6. ¹ H NMR (400MHz, DMSO-d ₆ ) δ 13.43 (br s, 1H), 8.31 (d, J=8.0Hz, 2H), 8.14-7.95 (m, 2H), 7.26 (d, J=7.5Hz) , 1H).

the fourth step:

To a solution of compound 6 (1.70 kg, 7.83 mol, 1 equiv) in t-butanol (12.7 L) was added 4A molecular sieves (850 g) and triethylamine (978.40 g, 9.67 mol, 1.35 L, 2 equiv), the above The mixture was stirred at 80°C for 1 hour. DPPA (1.33 kg, 4.83 mol, 1.05 L, 1 equiv) was then added dropwise to the reaction system and stirred at 80°C for 1 hour. The reaction mixture was cooled to 50°C, filtered, and the filter cake was washed with ethyl acetate (1.5 L*3). The filtrate was concentrated under reduced pressure, the obtained residue was dissolved in ethyl acetate (17 L), and the ethyl acetate was washed with water (17 L), aqueous citric acid (10%, 5 L) and saturated brine (7 L) in this order The organic phase was concentrated under reduced pressure to obtain compound 7.

the fifth step:

To a methanol solution (10.5 L) of compound 7 (2100 g, 4.21 mol, 1 equiv) was added a hydrogen chloride/methanol solution (4 mol/L, 10.5 L, 9.97 equiv), and the above mixture was stirred at 40°C for 4 hours . The reaction mixture was cooled to 20°C, filtered, and the filtrate was concentrated under reduced pressure. The obtained residue was dissolved in ethyl acetate (20 L) and water (15 L), and the organic phase after separation was adjusted to pH 7-8 with saturated aqueous sodium bicarbonate solution (7 L). The organic phase was washed with saturated brine (10 L) and concentrated under reduced pressure to obtain Compound 8.

Step 6:

To a solution of compound 8 (1470 g, 4.28 mol, 1 equiv) in dichloromethane (1470 mL) at 0 °C was added NBS (762.15 g, 4.28 mol, 1 equiv) portionwise, the resulting mixture was heated at 0 °C Stir for 1 hour. At 0 °C, saturated aqueous sodium sulfite solution (5 L) was added dropwise to the above reaction mixture, and stirred at 10 °C for 30 minutes, the layers were separated, and the obtained organic phase was sequentially water (10 L) and saturated brine (5 L) ) was washed and concentrated under reduced pressure. The obtained residue was dispersed in a mixed solvent (2.5 times the volume) of methyl tert-butyl ether:n-heptane=1:1.5, and stirred at 15°C for 5 hours. The above mixture was filtered, and the filter cake was washed with a mixed solvent of methyl tert-butyl ether:n-heptane=1:1.5 (0.2 L*2), and then dried under vacuum to obtain Compound 9.

Step 7:

To a mixed solution of compound 9 (330 g, 795.48 mol, 1 equiv) in ethanol (3000 mL) and water (1500 mL) was added ammonium chloride (212.76 g, 3.98 mol, 5 equiv), and the resulting reaction mixture was heated To 80°C, then iron powder (133.27 g, 2.39 mol, 3 equiv.) was added portionwise, and after the addition was complete, the reaction mixture was stirred at 80°C for 2 hours. The reaction mixture was cooled to 30°C and filtered, and the filtrate was concentrated under reduced pressure. The obtained residue was extracted with ethyl acetate (1.5 L*2), the organic phases were combined, washed with water (1 L) and saturated brine (1 L) successively, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure, Compound 10 was obtained.

Step 8:

To a solution of compound 10 (1140 g, 3.11 mol, 1 equiv) in N,N'-dimethylformamide (11.4 L) was added DPPF (34.52 g, 62.28 mmol, 0.02 equiv), zinc bromide (42.07 g) , 186.83 mmol, 9.35 mL, 0.06 equiv), zinc powder (20.36 g, 311.38 mmol, 0.1 equiv), zinc cyanide (219.38 g, 1.87 mol, 0.6 equiv) and Pd ₂ (dba) ₃ (28.51 g, 31.14 mmol, 0.01 equiv), the above mixture was heated to 100 °C under nitrogen and stirred for 16 h. Zinc powder (20.36 g, 311.38 mmol), DPPF (69.04 g, 124.56 mmol), Pd ₂ (dba) ₃ (57.02 g, 62.28 mmol) were added to the reaction system, and the above mixture was heated to 110 °C with stirring 15 hours. To the reaction system was added zinc powder (6 g, 91.76 mmol), DPPF (10 g, 19.84 mmol), zinc cyanide (50 g, 425.80 mmol), Pd ₂ (dba) ₃ (10 g, 10.92 mmol), the above mixture was further stirred at 110°C for 4 hours. The reaction mixture was cooled to 40°C and filtered. The filtrate was poured into a stirred aqueous sodium chloride solution (10%, 50 L). The mixture was filtered. The filter cake was dissolved in ethyl acetate (20 L) and filtered. The filtrate was Saturated brine (5 L) was used for washing, and the liquid was separated. To the resulting organic phase was added thiourea resin (750 g) and stirred at 50°C for 11 hours. The above mixture was filtered, and the filtrate was concentrated under reduced pressure to obtain compound 11. LCMS (ESI) m/z: 313.1 (M+1).

Step 9:

A solution of compound 11 (970 g, 2.48 mol, 1 equiv) in formic acid (4900 mL) was heated to 100°C, then concentrated sulfuric acid (607.75 g, 6.20 mol, 330.30 mL, 2.5 equiv) was added, and the resulting mixture was heated at 100°C Stir for 1 hour. The reaction solution was cooled to 20°C, poured into water (30 L), and the filter cake was collected after filtration to obtain a mixture of compound 12 and compound 13. Compound 12: LCMS (ESI) m/z: 369.1 (M+1); Compound 13: LCMS (ESI) m/z: 341.1 (M+1).

Step 10:

The above mixture of compound 12 and compound 13 (1080 g, 2.93 mol, 1 equiv.) was dissolved in a mixed solvent of ethanol (10 L) and water (5 L), and concentrated sulfuric acid (612.72 g, 6.25 mol, 333 mL) was added dropwise. , 2.13 equiv.), the above mixture was heated to 100 °C and stirred for 1 hour. The reaction mixture was concentrated under reduced pressure to remove ethanol, then water (10 L) was added to the residue, the pH of the solution was adjusted to 7 with aqueous sodium hydroxide solution (10%), the mixture was filtered, and the filter cake was washed with water (3 L) Vacuum drying gave crude product. The crude product was dispersed in a mixed solvent (4.5 L) of methanol/ethyl acetate=2/3, and the mixture was stirred at 20°C for 16 hours. After filtration, the filter cake was vacuum dried to obtain compound 13.

Step 11:

To a solution of compound 13 (490 g, 1.40 mol, 1 equiv) in N,N'-dimethylacetamide (1000 L) was added compound 14 (520.11 g, 2.79 mol, 2 equiv), N,N'-bis Isopropylethylamine (541.37 g, 4.19 mol, 729.61 mL, 3 equiv) and PyBrOP (846.19 g, 1.82 mol, 1.3 equiv), the resulting mixture was stirred at 25°C for 16 hours. With stirring, the reaction mixture was poured into water (20 L), stirred at 15°C for 1 hour, filtered, and the filter cake was washed with water (5 L*2) and dried in vacuo. The obtained crude product was dispersed in a mixed solvent (6 liters) of methyl tert-butyl ether/petroleum ether=1/1, and the mixture was stirred at 15°C for 4 hours. After filtration, the filter cake was dried under vacuum to obtain compound 15. ¹ H NMR (400MHz, DMSO-d ₆ ) δ 8.85-8.69 (m, 1H), 7.22-7.03 (m, 2H), 6.57 (d, J=8.4Hz, 1H), 6.47-6.32 (m, 1H) ), 5.70(s, 2H), 3.77(br t, J=5.0Hz, 4H), 3.55(br s, 4H), 1.49-1.41(m, 9H).

Step 12:

To a solution of compound 15 (720 g, 1.36 mol, 1 equiv) in acetonitrile (7.2 L) was added NCS (398.97 g, 2.99 mol, 2.2 equiv) and the mixture was stirred at 75°C for 2 hours. The above mixture was stirred at 80°C for 2 hours. Additional NCS (90.70 g, 679.26 mmol, 0.5 equiv) was added to the above reaction solution and stirred at 80°C for 1 hour. The reaction mixture was cooled to 40°C, water (29 L) was added to the reaction system, the resulting mixture was filtered, and the filter cake was washed with water (5 L) and dried in vacuo to give compound 16. ¹ H NMR (400MHz, CD ₃ OD) δ 8.78(s, 1H), 7.56(d, J=7.5Hz, 1H), 7.26(s, 1H), 4.03-3.86(m, 4H), 3.66(br s, 4H), 1.52 (s, 9H).

Step Thirteen:

To a solution of compound 16 (760 g, 1.18 mol, 1 equiv) in ethyl acetate (3.8 L) was added hydrogen chloride/ethyl acetate solution (4 mol/L, 3.8 L, 12.85 equiv), and the above reaction solution was heated at 15°C under stirring for 2 hours. The above reaction mixture was filtered, and the filter cake was washed successively with ethyl acetate (2 L*2) and petroleum ether (2 L) and then dried under vacuum to obtain the hydrochloride salt of compound 17. ¹ H NMR (400MHz, DMSO-d ₆ ) δ 9.91 (br s, 2H), 9.04-8.84 (m, 1H), 7.74 (d, J=7.5Hz, 1H), 7.29 (s, 1H), 4.23 -4.11 (m, 2H), 4.06-4.01 (m, 2H), 3.29 (br d, J=3.7Hz, 4H).

Step 14:

To a solution of the hydrochloride salt of compound 17 (690 g, 1.24 mol, 1 equiv) in tetrahydrofuran (4850 mL) and water (2150 mL) was added N,N'-diisopropylethylamine (479.49 g, 3.71 mol, 646.22 mL, 3 equiv), the resulting mixture was cooled to 0 °C, then compound 18 (111.93 g, 1.24 mol, 100.84 mL, 1 equiv) was added dropwise to the reaction system over about 30 minutes, and the above mixture was cooled at 0 °C Stir for 30 minutes. To the above reaction mixture was added water (20 L) with stirring, filtered and the filter cake was dried in vacuo. The obtained residue was purified by silica gel column chromatography (dichloromethane/methanol=30/1 to 10/1, volume ratio) to obtain a racemate of the target product. The racemate was purified by SFC (column: DAICEL CHIRALCEL OD (250mm*30mm, 10μm); mobile phase A: carbon dioxide; mobile phase B: methanol with 0.1% ammonia; 50%-50%, 2.55min; 1255min) , the obtained crude product (retention time = 1.88 min) was dispersed in methyl tert-butyl ether (2 L) and stirred at 15°C for 16 hours. The mixture was filtered, the filter cake was washed with methyl tert-butyl ether (100 mL*2), and the compound of formula (I) was obtained after vacuum drying. ¹ H NMR (400MHz, DMSO-d ₆ ) δ 8.81 (s, 1H), 7.72 (d, J=7.5Hz, 1H), 7.18 (s, 1H), 6.83 (dd, J=10.4, 16.7Hz, 1H), 6.33-6.10 (m, 3H), 5.83-5.69 (m, 1H), 3.99-3.65 (m, 8H).

Example 2: Preparation of the crystal form of compound A of formula (I)

A solution of the compound of formula (I) (2.01 g, 3.59 mmol, 1 equiv) in ethyl acetate (10 mL) was stirred at 60°C for 16 hours. The reaction mixture was filtered, and the filter cake was washed with ethyl acetate (5 mL), and dried to obtain a crystal form of compound A of formula (I).

The XRPD spectrum of the crystal form of compound A of formula (I) is shown in FIG. 1 , the DSC spectrum is shown in FIG. 2 , and the TGA spectrum is shown in FIG. 3 .

Example 3: Preparation of compound B crystal form of formula (II)

A solution of the compound of formula (I) (1 g, 1.83 mmol, 1 equiv) and methanesulfonic acid (175.55 mg, 1.83 mmol, 130.04 μl, 1 equiv) in acetonitrile (10 mL) was stirred at 20°C for 16 hours , with solid precipitation. Filtration to obtain compound B crystal form of formula (II). ¹ H NMR (400MHz, CD ₃ OD) δ 8.86 (s, 1H), 7.62 (d, J=7.46 Hz, 1H), 7.48 (s, 1H), 6.82 (dd, J=16.75, 10.64 Hz, 1H ), 6.33(dd, J=16.81, 1.90Hz, 1H), 5.89-5.80(m, 1H), 4.46-4.29(m, 4H), 4.07-3.91(m, 4H), 2.66(s, 3H); LCMS (ESI) m/z: 531.1 (M+1).

The XRPD spectrum of the crystal form of compound B of formula (II) is shown in Fig. 4 , the DSC spectrum is shown in Fig. 5 , and the TGA spectrum is shown in Fig. 6 .

Example 4: Preparation of formula (I) compound p-toluenesulfonate salt form C

A solution of the compound of formula (I) (1 g, 1.83 mmol, 1 equiv) and p-toluenesulfonic acid (314.56 mg, 1.83 mmol, 1 equiv) in acetonitrile (10 mL) was stirred at 20°C for 16 hours, a solid Precipitate. Filtration to obtain the crystal form C of the compound of formula (I) p-toluenesulfonate. ¹ H NMR (400MHz, CD ₃ OD) δ 8.93-8.75 (m, 1H), 7.63 (dd, J=14.31, 7.82Hz, 3H), 7.46 (s, 1H), 7.23 (d, J=7.95Hz) ,2H),6.80(dd,J＝16.75,10.64Hz,1H),6.32(dd,J＝16.81,1.90Hz,1H),5.79-5.95(m,1H),4.35(br d,J＝5.01Hz , 4H), 3.97 (br d, J=11.98Hz, 4H), 2.44-2.32 (m, 3H).

The XRPD spectrum of the compound of formula (I) p-toluenesulfonate salt form C is shown in Figure 7, the DSC spectrum is shown in Figure 8, and the TGA spectrum is shown in Figure 9.

Example 5: Preparation of Compound D of Formula (I) Hydrochloride Crystal Form

To a solution of the compound of formula (I) (1 g, 1.83 mmol, 1 equiv) in acetonitrile (10 mL) was added hydrochloric acid (10 mol/L, 200.94 μL, 35% pure, 1.1 equiv) and the resulting mixture was placed in After stirring at 15°C for 16 hours, a solid was precipitated. Filtration to obtain the hydrochloride salt form D of the compound of formula (I). ¹ H NMR (400MHz, DMSO-d ₆ )δ8.80-9.10(m,1H),7.77(d,J=7.50Hz,1H),7.34(s,1H),6.81-6.95(m,1H), 6.33-6.73 (m, 2H), 6.15-6.28 (m, 1H), 5.66-5.87 (m, 1H), 3.61-4.22 (m, 8H).

The XRPD spectrum of the crystalline form D of the compound of formula (I) hydrochloride is shown in Figure 10, the DSC spectrum is shown in Figure 11, and the TGA spectrum is shown in Figure 12.

Example 6: Study on the hygroscopicity of the crystal form

Experimental conditions:

Instrument model: SMS DVS Advantage dynamic vapor adsorption instrument

Test conditions: Take a sample (10-15 mg) and place it in the DVS sample tray for testing.

The detailed DVS parameters are as follows:

Temperature: 25℃

Balance: dm/dt=0.01%/min (minimum: 10min, longest: 180min)

Drying: 120min at 0%RH

RH (%) test rung: 10%

RH (%) test step range: 0%-90%-0%

The hygroscopicity evaluation classification is shown in Table 5.

table 5

Hygroscopic classification	Moisture gain weight*
deliquescence	Absorbs enough water to form a liquid
Very hygroscopic	ΔW%≥15%
hygroscopic	15%>ΔW%≥2%
slightly hygroscopic	2%>ΔW%≥0.2%
No or almost no hygroscopicity	ΔW%<0.2%

*Wetting weight gain at 25±1°C and 80±2% RH.

Experimental results:

Under the conditions of 25°C and 80% RH, the hygroscopic weight gain of compound B of formula (II) is 1.53%, and the hygroscopic weight gain of compound of formula (I) hydrochloride crystalline form D is 3.54%; the compound of formula (I) The hygroscopic weight gain of p-toluenesulfonate salt form C was 2.48%.

Experimental results:

The crystal form of compound B of formula (II) is slightly hygroscopic.

Example 7: Stability test of crystal form

Experimental operation:

According to the influencing factors and accelerated test conditions, accurately weigh 5 mg of compound B crystal form of formula (II), place in duplicate at the bottom of a 40 mL glass sample bottle, spread it out into a thin layer, and place it under appropriate conditions. In the open condition, make some small holes in the aluminum foil to ensure that the sample can be fully contacted with the ambient air. In addition, a small amount of samples were taken and placed in 40mL glass sample bottles, and the crystal state was to be determined under the same conditions. At the inspection time point, the corresponding test samples were taken out, covered with bottle caps, and the 0-day-old samples were taken out of the refrigerator, and analyzed after the samples returned to room temperature. About 10 mg of the test article is used for XRPD detection. The compounds to be investigated were placed under the following conditions and samples were taken at different time points to detect the content and related substances. The research conditions and test items are shown in Table 6.

Table 6

Note: Test item X includes: content and related substances; X* refers to the test items of the 0-day sample (including content and related substances), and X* will be used as a reference for sampling and testing at different time points in the later period; N/A means no sampling.

Experimental results: The experimental results are shown in Table 7.

Table 7

Note: N/A means not detected.

Experimental conclusion: The crystal form of compound B of formula (II) has good stability.

Example 8: Solubility experiment of crystalline form simulating gastrointestinal fluid

Experimental operation:

Weigh 2-3 mg of compound B crystal form of formula (II) and compound A crystal form of formula (I) into a 1.5 mL liquid phase vial, add 1 mL of medium, and place at 37° C. and stir on a 700 rpm magnetic stirrer. Samples were taken after 2 hours, pH was determined, and concentration was determined using HPLC.

Experimental results: The experimental results are shown in Table 8.

Table 8

Solubility (mg/mL)_2 hours	FaSSIF	FeSSIF	SGF	water
Formula (II) compound B crystal form	0.041	0.409	0.366	0.073
Crystal form of compound A of formula (I)	0.029	0.181	0.030	0.008

Note: FaSSIF: simulates the intestinal fluid in the small intestine in the state of human pre-prandial starvation;

FeSSIF: simulates the intestinal fluid in the small intestine of humans in a state of satiety after a meal;

SGF: Simulates the gastric juice of empty stomach in human starvation state;

Experimental results:

The crystal form of compound B of formula (II) has better solubility, which is about 9 times higher than that of crystal form of compound A of formula (I) in water.

Example 9: Solubility test of compound B crystal form of formula (II) in different pH media

Experimental operation:

About 2 mg of compound B crystal form of formula (II) was added to a 2 ml glass bottle, 9 portions were weighed, and then 1 ml of different media (0.1mol/L HCl, 0.01mol/L HCl, water, pH3. 8. pH4.5, pH5.5, pH6.0, pH6.8, pH7.4 buffer solution), add the magnetron to the above suspension, place it on a magnetic stirrer (the temperature is 37°C, avoid light ). After stirring for 24 hours, the samples were sampled and centrifuged, the upper layer sample was filtered through a filter membrane, and the filtrate was assayed for its concentration and pH value by HPLC.

Experimental results: The experimental results are shown in Table 9.

Table 9

solvent	pH (24 hours)	Status (24 hours)	Solubility (mg/mL)_24 hours
0.1mol/L HCl	0.98	Suspended	0.98
0.01mol/L HCl	1.79	Suspended	0.11
pH3.8	3.40	Suspended	0.016
pH4.5	4.31	Suspended	0.014
pH5.5	5.26	Suspended	0.014

pH6.0	5.47	Suspended	0.013
pH6.8	6.55	Suspended	0.013
pH7.4	7.15	Suspended	0.012
water	2.34	Suspended	0.038

Experimental results:

The stronger the acidity, the higher the solubility of the compound B crystal form of formula (II).

Example 10: Confirmation of the stereo configuration of the compound of formula (I)

Preparation of crystals of the compound of formula (I): The crystals of the compound of formula (I) are obtained by culturing at room temperature for 10 days under the condition of ethanol using a solvent evaporation method.

Experimental results: the ellipsoid diagram of the three-dimensional structure of the compound of formula (I) is shown in Figure 13, which is the S configuration. The crystal structure data and parameters of the compound of formula (I) are shown in Tables 10-1, 10-2, 10-3 and 10-4.

Table 10-1 Crystal structure data and determination parameters of the compound of formula (I)

Table 10-2 Atomic coordinates (×10 ⁴ ) and equivalent isotropic shift parameters of compound crystals of formula (I)

/	x	y	z	U(eq)
Cl(1)	4701(1)	-2402(2)	2651(3)	66(1)

Cl(2)	4826(1)	3346(2)	2644(2)	58(1)
F(1)	4216(1)	1535(4)	8150(5)	45(1)
F(2)	4158(1)	-739(4)	7878(4)	45(1)
F(3)	3845(1)	321(5)	9849(3)	48(1)
F(4)	4006(2)	3249(5)	4384(6)	68(1)
O(1)	3238(1)	914(4)	2351(4)	32(1)
O(2)	1607(2)	-1111(4)	12986(6)	48(1)
N(1)	2026(1)	2154(5)	5751(5)	29(1)
N(2)	2467(1)	1878(4)	3421(5)	28(1)
N(3)	3562(1)	742(4)	5165(4)	24(1)
N(4)	2220(1)	1289(4)	8539(5)	24(1)
N(5)	1573(1)	-39(4)	10314(6)	35(1)
N(6)	3921(1)	-1815(4)	4361(5)	21(1)
C(1)	2325(1)	1487(5)	6890(5)	22(1)
C(2)	2117(2)	2303(6)	4113(6)	32(1)
C(3)	2786(1)	1313(5)	4599(5)	23(1)
C(4)	3199(1)	971(5)	3904(5)	24(1)
C(5)	3521(1)	631(5)	6955(5)	23(1)
C(6)	3131(1)	760(4)	7558(5)	22(1)
C(7)	2745(1)	1097(4)	6374(5)	21(1)
C(8)	1854(1)	2108(5)	9111(6)	26(1)
C(9)	1454(2)	1172(5)	9171(7)	33(1)
C(10)	1928(2)	-905(5)	9751(7)	33(1)
C(11)	2335(1)	-2(5)	9584(5)	25(1)
C(12)	1441(2)	-237(5)	11915(8)	40(1)
C(13)	1061(2)	635(7)	12349(10)	62(1)
C(14)	681(3)	565(11)	11662(15)	87(1)
C(15)	3976(1)	674(5)	4465(5)	26(1)
C(16)	4138(2)	-630(6)	4023(6)	32(1)
C(17)	4514(2)	-742(7)	3212(7)	38(1)
C(18)	4726(2)	494(7)	2807(6)	38(1)
C(19)	4561(2)	1813(6)	3216(6)	35(1)
C(20)	4177(2)	1957(5)	4051(6)	30(1)
C(21)	3935(1)	441(6)	8208(5)	30(1)
H(6A)	3679	-1755	4858	25
H(6B)	4021	-2658	4086	25
H(2B)	1900	2783	3334	38
H(6C)	3110	629	8771	26
H(8A)	1779	2913	8291	31
H(8B)	1947	2513	10291	31
H(9A)	1217	1730	9620	40
H(9B)	1344	831	7976	40
H(10A)	1826	-1353	8605	40
H(10B)	2006	-1680	10614	40
H(11A)	2475	277	10767	30
H(11B)	2550	-575	9018	30
H(13A)	1122	1320	13261	75
H(14A)	601	-101	10743	104
H(14B)	465	1176	12047	104
H(18A)	4985	440	2249	46

Table 10-3 Bond Lengths of Compound Crystals of Formula (I)

and bond angle [deg]

Table 10-4 Torsion angle [deg] of compound crystals of formula (I)

Experimental Example 1: Cell Experiment

Purpose:

The purpose of this experiment is to verify the proliferation inhibitory effect of the compounds of the present invention on KRAS G12C-mutated NCI-H358 human non-small cell lung cancer cells, KRAS G12C-mutated MIA PaCa2 human pancreatic cancer cells and wild-type A375 human malignant melanoma cells. Main reagents:

Cell line NCI-H358, cell line A375, cell line MIA Paca2, Cell Titer-Glo detection kit, RPMI1640 medium, DMEM cell medium, fetal bovine serum, 0.25% trypsin-EDTA digestion solution, DPBS, cell culture grade DMSO, penicillin

Main instruments:

Multi-labeled microplate detector Envision, cell culture flask, 384 cell culture microplate, Vi-cell XR cell viability analyzer, CO2 incubator, 300μL 12-channel electric pipette, Echo ultrasonic nano-upgrade liquid workstation

experimental method:

Add 40 μL of phosphate buffer to the peripheral wells of three 384-well plates, respectively, and add 40 μL of the cell suspension to be tested to the other wells of each plate (plate 1: NCI-H358 cell suspension, which contains 500 cells) NCI-H358 cells; Plate 2: MIA PaCa2 cell suspension containing 300 MIA PaCa2 cells; Plate 3: A375 cell suspension containing 300 A375 cells). The three cell plates were then placed in a carbon dioxide incubator overnight. The compounds to be tested were diluted 3-fold with Echo, and the compounds were diluted in 10 concentration gradients (diluted from 50 μM to 0.003 μM) and added 100 nl to the corresponding wells of the cell plate respectively. After drug addition, rows A, P, 1, 24 Add 40 μL of phosphate buffer to each well of the column, then place the cell plate back into the carbon dioxide incubator for 5 days. Add 20 μl of Promega CellTiter-Glo reagent per well to the cell plate, shake at room temperature in the dark for 10 minutes to stabilize the luminescence signal. Readings were performed on a PerkinElmer Envision multi-label analyzer.

Data analysis: _IC50 results were analyzed by GraphPad Prism 5.0 software from IDBS Corporation.

Experimental results:

The data of the antiproliferative activity _IC50 of the compounds of the present invention on NCI-H358 (G12C mutant) cells, A375 (wild type) cells and MIA PaCa2 (G12C mutant) cells are shown in Table 11-1.

Table 11-1

test compound	NCI-H358 IC 50 (μM)	A375 IC 50 (μM)	MIA PaCa2 IC 50 (μM)
Compounds of formula (I)	0.05	7.32	0.07

Experimental results:

The compound of the present invention shows high cell anti-proliferation activity on KRAS G12C mutant cells NCI-H358 and MIA PaCa2, and at the same time has weak anti-proliferative activity on wild-type A375 cells, showing high selectivity.

Experimental Example 2: Pharmacokinetic Evaluation Experiment in Rats

Purpose:

Male SD rats were used as the test animals, and LC/MS/MS method was used to determine the drug concentrations in the plasma of rats at different times after intravenous and intragastric administration of the test compounds. To study the pharmacokinetic behavior of test compounds in rats, and to evaluate their pharmacokinetic characteristics.

Experimental scheme: Experimental animals: 10 healthy adult male SD rats were divided into 4 groups according to the principle of similar body weight, with 2 rats in each group of IV group (two groups) and 3 rats in each group of PO group (two groups). Animals were purchased from Beijing Weitong Lihua Laboratory Animal Co., Ltd.

Drug preparation:

Group IV: Weigh an appropriate amount of sample, add appropriate amount of DMSO, PEG400 and water in turn according to the volume ratio of 10:60:30, and then stir and ultrasonicate to reach a clear state of 1.5 mg/mL.

PO group: Weigh an appropriate amount of sample, add an appropriate amount of DMSO, PEG400 and water in turn according to the volume ratio of 10:60:30, and after stirring and ultrasonicating, it reaches a clear state of 1.0 mg/mL.

Dosing:

After an overnight fast, the IV group was administered intravenously with a volume of 2 mL/kg and a dose of 3 mg/kg; the PO group was administered by intragastric administration with a volume of 10 mL/kg and a dose of 10 mg/kg.

Experimental operation:

Male SD rats in the intravenous injection group were given the test compounds, and 200 μL of blood was collected at 0.0833, 0.25, 0.5, 1, 2, 4, 6, 8, and 24 hours, and placed in a commercial pre-filled EDTA-K ₂ in the anticoagulant tube. After the test compound was administered to the gavage group, 200 μL of blood was collected at 0.25, 0.5, 1, 2, 4, 6, 8, and 24 hours, respectively, and placed in a commercial anticoagulation tube pre-filled with EDTA-K ₂ middle. The tubes were centrifuged for 15 minutes to separate plasma and stored at -60°C. Animals were allowed to eat 2 hours after dosing. The content of the test compounds in the plasma of rats after intravenous and intragastric administration was determined by LC/MS/MS. The linear range of the method was 2.00-6000 nM; plasma samples were analyzed after acetonitrile precipitation of proteins.

Experimental results: The experimental results are shown in Table 11-2.

Table 11-2

Note: Cl: clearance; _Vd : volume of distribution; AUC: exposure; T1 _/2 : half-life; _Cmax : maximum compound concentration after oral administration; _Tmax : time to reach _Cmax ; F: bioavailability Spend.

Experimental results:

In the rat pharmacokinetic evaluation experiment, the compound of the present invention showed higher exposure and better oral availability than the reference compound ARS-1620.

Experimental Example 3: In vivo efficacy test (1)

Purpose:

The in vivo efficacy of the test compounds on the human non-small cell lung cancer NCI-H358 subcutaneous xenograft tumor model was evaluated.

Experimental operation:

BALB/c nude mice, female, 6-8 weeks old, weighing 18-21 grams. A total of 100 are required. Provided by Shanghai Lingchang Laboratory Animal Co., Ltd. The NCI-H358 tumor cells were resuspended in PBS, prepared into 0.1 mL (5×10 ⁶ cells) of cell suspension, and subcutaneously inoculated into the right back of each mouse (5×10 ⁶ /mice) to wait for tumor growth . Randomization was started when the average tumor volume reached about 150-200 mm ³ , and the doses were shown in Table 12. Tumor diameters were measured with vernier calipers twice a week. The calculation formula of tumor volume is: V=0.5a×b ² , a and b represent the long and short diameters of the tumor, respectively. The tumor-inhibitory efficacy of the compounds was evaluated by TGI (%). TGI (%), reflecting tumor growth inhibition rate. Calculation of TGI (%): TGI (%)=[(1-(average tumor volume at the end of administration of a certain treatment group - average tumor volume at the beginning of administration of this treatment group)/(average tumor volume at the end of treatment in the solvent control group) - Average tumor volume at the start of treatment in the solvent control group)] × 100%.

Experimental results: The experimental results are shown in Table 12.

Table 12

Experimental results:

The compounds of the present invention show good in vivo efficacy in the human non-small cell lung cancer NCI-H358 subcutaneous xenograft tumor model. Twenty days after the start of administration, the compounds of the present invention had stronger antitumor effects than the reference compound ARS-1620.

Experimental Example 4: In vivo efficacy test (2)

Purpose:

The in vivo efficacy of test compounds was evaluated in a human pancreatic cancer x-MIA-PaCa2 cell subcutaneous xenograft tumor model.

Experimental operation:

NU/NU mice, female, 6-8 weeks old, weighing 17-20 grams. A total of 100 animals are required (30% more animals vaccinated). Provided by Beijing Weitong Lihua Technology Co., Ltd. 0.2mL (10×10 ⁶ cells) x-MIA-PaCa2 cells (plus Matrigel, volume ratio of 1:1) were subcutaneously inoculated into the right back ^of each mouse, and the grouping started when the average tumor volume reached about 150mm Administration, the administration doses are shown in Table 13. Tumor diameters were measured with vernier calipers twice a week. The calculation formula of tumor volume is: V=0.5a×b ² , a and b represent the long and short diameters of the tumor, respectively. The tumor-inhibitory efficacy of the compounds was evaluated by TGI (%). TGI (%), reflecting tumor growth inhibition rate. Calculation of TGI (%): TGI (%)=[(1-(average tumor volume at the end of administration of a certain treatment group - average tumor volume at the beginning of administration of this treatment group)/(average tumor volume at the end of treatment in the solvent control group) - Average tumor volume at the start of treatment in the solvent control group)] × 100%.

Experimental results: The experimental results are shown in Table 13.

Table 13

group	Tumor volume (mm 3 ) (day 14)	TGI(%)
solvent control	1670	--
ARS-1620(50mg/kg)	907	50.36
Compound of formula (I) (50mg/kg)	204	96.77

Experimental results:

The compounds of the present invention show good in vivo efficacy in human pancreatic cancer x-MIA-PaCa2 cell subcutaneous xenograft tumor model. 14 days after the start of administration, the compound of the present invention has stronger antitumor effect than the reference compound ARS-1620.

Experimental Example 5: In vivo efficacy test (3)

Purpose:

In this experiment, the anti-tumor effect of compound B crystal form of formula (II) was evaluated by using the human non-small cell lung cancer NCI-H358 subcutaneous xenograft tumor nude mouse model.

Experimental operation:

BALB/c nude mice, female, 6-8 weeks old, weighing 17-22 grams. A total of 40 mice are required (excluding the remaining mice in the group). Provided by Beijing Weitong Lihua Technology Co., Ltd. Each animal was inoculated with 0.1 mL (5×10 ⁶ cells) of human non-small cell lung cancer NCI-H358 cells in the right back position. When the average tumor volume reached 168 mm ³ , random grouping was adopted, and the administration was started. The dosage is shown in Table 14. shown. Tumor diameters were measured with vernier calipers twice a week. The calculation formula of tumor volume is: V=0.5a×b ² , a and b represent the long and short diameters of the tumor, respectively. The tumor-inhibitory efficacy of the compounds was evaluated by TGI (%). TGI (%), reflecting tumor growth inhibition rate. Calculation of TGI (%): TGI (%)=[(1-(average tumor volume at the end of administration of a certain treatment group - average tumor volume at the beginning of administration of this treatment group)/(average tumor volume at the end of treatment in the solvent control group) - Average tumor volume at the start of treatment in the solvent control group)] × 100%.

Experimental results: The experimental results are shown in Table 14.

Table 14

group	Tumor volume (mm 3 ) a (day 23)	TGI (%) (Day 23)
solvent control	1169±152	--
ARS-1620(20mg/kg)	303±52	85.8
Crystal form of compound B of formula (II) (1.5mg/kg)	566±120	60.2
Formula (II) Compound B Crystal Form (3mg/kg)	332±119	83.6
Crystal form of compound B of formula (II) (20mg/kg)	114±63	105.4

Note: a. Data are expressed as "mean ± standard error";

Experimental results:

The compounds of the present invention can dose-dependently inhibit the growth of subcutaneously transplanted tumors of human lung cancer NCI-H358 cells in tumor-bearing mice, show significant antitumor activity, and are well tolerated by animals. Compared with the control compound ARS-1620 at the same dose, the compound of the present invention has better antitumor activity.

Experimental Example 6: In vivo efficacy test (4)

Purpose:

In this experiment, human lung cancer LU-01-0030 subcutaneous xenograft tumor nude mice model was used to evaluate the antitumor effect of compound B crystal form of formula (II).

Experimental operation:

BALB/c nude mice, female, 6-8 weeks old, weighing 15-21 grams. A total of 40 mice are required (excluding the remaining mice in the group). Provided by Beijing Weitong Lihua Technology Co., Ltd. Each animal was inoculated with LU-01-0030FP4 tumor mass with a volume of about 30 mm ³ on the right back. When the average tumor volume reached 199 mm ³ , random grouping was adopted and the administration was started. The dosage is shown in Table 15. Tumor diameters were measured with vernier calipers twice a week. The calculation formula of tumor volume is: V=0.5a×b ² , a and b represent the long and short diameters of the tumor, respectively. The tumor-inhibitory efficacy of the compounds was evaluated by TGI (%). TGI (%), reflecting tumor growth inhibition rate. Calculation of TGI (%): TGI (%)=[(1-(average tumor volume at the end of administration of a certain treatment group - average tumor volume at the beginning of administration of this treatment group)/(average tumor volume at the end of treatment in the solvent control group) - Average tumor volume at the start of treatment in the solvent control group)] × 100%.

Experimental results: The experimental results are shown in Table 15.

Table 15

group	Tumor volume (mm 3 ) a (Day 27)	TGI (%) (Day 27)
solvent control	832±87	--
ARS-1620(200mg/kg)	186±23	102.1
Crystal form of compound B of formula (II) (5mg/kg)	281±28	87.0
Crystal form of compound B of formula (II) (15mg/kg)	142±14	109.2
Formula (II) Compound B Crystal Form (45mg/kg)	79±15	119.0

Note: a. Data are expressed as "mean ± standard error";

Experimental results:

The compounds of the present invention (5, 15, 45 mg/kg, PO, QD×4W) can dose-dependently inhibit the growth of human lung cancer LU-01-0030 xenograft mouse model tumor, show significant anti-tumor activity, and Animals tolerated well.

Experimental Example 7: In vivo efficacy test (5)

Purpose:

To evaluate the antitumor effect of compound B crystal form of formula (II) in human pancreatic cancer MIA PaCa-2 cell line subcutaneous xenograft BALB/c nude mouse animal model.

Experimental operation:

BALB/c nude mice, female, 6-7 weeks old, weighing 15.4-21.9 g. A total of 72 mice (48 plus 24 surplus mice) were required. Provided by Jiangsu Jicui Yaokang Biotechnology Co., Ltd. Each animal was subcutaneously inoculated with 5×10 ⁶ MIA PaCa-2 cells on the right side. When the average tumor volume reached 124 mm ³ , the animals were randomly divided into groups and started to be administered. The doses are shown in Table 16. Tumor diameters were measured with vernier calipers twice a week. The calculation formula of tumor volume is: V=0.5a×b ² , a and b represent the long and short diameters of the tumor, respectively. The tumor-inhibitory efficacy of the compounds was evaluated by TGI (%). TGI (%), reflecting tumor growth inhibition rate. Calculation of TGI (%): TGI (%)=[(1-(average tumor volume at the end of administration of a certain treatment group - average tumor volume at the beginning of administration of this treatment group)/(average tumor volume at the end of treatment in the solvent control group) - Average tumor volume at the start of treatment in the solvent control group)] × 100%.

Experimental results: The experimental results are shown in Table 16.

Table 16

group	Tumor volume (mm 3 ) a (Day 27)	TGI (%) (Day 27)
solvent control	1261.63±132.77	--
ARS-1620(15mg/kg)	654.18±88.64	53
ARS-1620(45mg/kg)	364.84±63.75	79
Crystal form of compound B of formula (II) (5mg/kg)	400.94±63.53	76
Crystal form of compound B of formula (II) (15mg/kg)	70.11±8.38	105
Formula (II) Compound B Crystal Form (45mg/kg)	30.7±2.1	108

Note: a. Data are expressed as "mean ± standard error";

Experimental results:

The compound of the present invention has a significant inhibitory effect on the tumor growth of MIA PaCa-2 human pancreatic cancer tumor model in a dose-dependent manner; and at the same dose, the compound of the present invention can significantly inhibit the growth of MIA PaCa-2 human pancreatic cancer. Better than ARS-1620.

Claims (35)

1. The crystal form of compound A of formula (I), its X-ray powder diffraction pattern has characteristic diffraction peaks at the following 2θ angles: 9.96±0.20°,

   19.83±0.20°, 22.23±0.20°.

   
2. The crystal form A according to claim 1, its X-ray powder diffraction pattern has characteristic diffraction peaks at the following 2θ angles: 9.96±0.20°, 18.09±0.20°, 19.83±0.20°, 22.23±0.20°, 22.65±0.20 °, 24.80±0.20°, 25.78±0.20°, 27.42±0.20°.
3. The crystal form A according to claim 2, its X-ray powder diffraction pattern has characteristic diffraction peaks at the following 2θ angles: 5.921°, 9.960°, 11.718°, 15.071°, 15.584°, 17.401°, 18.089°, 18.954° , 19.826°, 22.231°, 22.646°, 23.377°, 24.796°, 25.783°, 27.417°, 29.860°, 30.334°, 30.649°, 32.095°, 33.691°, 35.187°, 38.757°, 39.073°.
4. The crystal form A according to claim 3, its X-ray powder diffraction pattern is shown in Figure 1.
5. The crystal form A according to any one of claims 1 to 4, wherein the differential scanning calorimetry curve has an onset of an endothermic peak at 249.10±3°C.
6. Form A according to claim 5, its DSC spectrum is shown in Figure 2.
7. The crystal form A according to any one of claims 1 to 4 has a weight loss of 0.05409% in its thermogravimetric analysis curve at 120.17±3°C.
8. Form A according to claim 7, its TGA spectrum is shown in Figure 3.
9. Compounds of formula (II)

   
10. The crystal form of compound B of formula (II) has characteristic diffraction peaks at the following 2θ angles in its X-ray powder diffraction pattern: 10.16±0.20°, 17.75±0.20°, 24.32±0.20°.
11. The crystal form B according to claim 10, its X-ray powder diffraction pattern has characteristic diffraction peaks at the following 2θ angles: 10.16±0.20°, 17.75±0.20°, 19.25±0.20°, 19.98±0.20°, 22.59±0.20 °, 23.06±0.20°, 24.32±0.20°, 30.71±0.20°.
12. The crystal form B according to claim 11, its X-ray powder diffraction pattern has characteristic diffraction peaks at the following 2θ angles: 4.105°, 8.800°, 10.164°, 11.633°, 13.060°, 14.973°, 15.324°, 16.274° , 16.613°, 17.102°, 17.752°, 18.107°, 19.034°, 19.254°, 19.983°, 20.912°, 22.586°, 23.060°, 24.322°, 24.698°, 25.681°, 26.157°, 27.216°, 27.67° °, 28.627°, 29.353°, 30.277°, 30.711°, 32.386°, 33.335°, 34.139°, 35.01°, 35.448°, 35.859°, 36.608°, 37.976°.
13. The crystal form B according to claim 12, its X-ray powder diffraction pattern is shown in FIG. 4 .
14. The crystal form B according to any one of claims 10 to 13, wherein the differential scanning calorimetry curve has an onset of an endothermic peak at 253.00±3°C.
15. The crystal form B according to claim 14, its DSC spectrum is shown in Figure 5.
16. The crystal form B according to any one of claims 10 to 13, whose thermogravimetric analysis curve has a weight loss of 0.4116% at 150.14±3°C.
17. The crystal form B according to claim 16, its TGA spectrum is shown in Figure 6.
18. The compound of formula (I) p-toluenesulfonate salt form C, its X-ray powder diffraction pattern has characteristic diffraction peaks at the following 2θ angles: 19.57±0.20°, 21.56±0.20°, 24.01±0.20°.
19. The crystal form C according to claim 18, its X-ray powder diffraction pattern has characteristic diffraction peaks at the following 2θ angles: 12.17±0.20°, 15.78±0.20°, 16.69±0.20°, 19.57±0.20°, 21.56±0.20 °, 24.01±0.20°, 25.02±0.20°, 34.80±0.20°.
20. The crystal form C according to claim 19, its X-ray powder diffraction pattern has characteristic diffraction peaks at the following 2θ angles: 8.030°, 12.169°, 13.194°, 14.399°, 15.332°, 15.779°, 16.055°, 16.496° , 16.687°, 18.996°, 19.289°, 19.567°, 20.556°, 20.987°, 21.562°, 21.897°, 22.864°, 24.006°, 24.281°, 25.018°, 26.433°, 28.397°, 28.81.6°, 30.430° °, 31.773°, 31.935°, 34.795°.
21. The crystal form C according to claim 20, its X-ray powder diffraction pattern is shown in FIG. 7 .
22. The crystal form C according to any one of claims 18 to 21, wherein the differential scanning calorimetry curve has an onset of an endothermic peak at 262.38±3°C.
23. The crystal form C according to claim 22, its DSC spectrum is shown in Figure 8.
24. The crystal form C according to any one of claims 18 to 21, whose thermogravimetric analysis curve has a weight loss of 1.542% at 146.27±3°C.
25. The crystal form C according to claim 24, its TGA spectrum is shown in Figure 9.
26. The compound of formula (I) hydrochloride salt form D, its X-ray powder diffraction pattern has characteristic diffraction peaks at the following 2θ angles: 17.50±0.20°, 24.50±0.20°, 25.21±0.20°.
27. The crystal form D according to claim 26, its X-ray powder diffraction pattern has characteristic diffraction peaks at the following 2θ angles: 16.39±0.20°, 17.50±0.20°, 19.49±0.20°, 19.96±0.20°, 22.09±0.20 °, 24.50±0.20°, 25.21±0.20°, 27.85±0.20°.
28. The crystal form D according to claim 27, its X-ray powder diffraction pattern has characteristic diffraction peaks at the following 2θ angles: 6.742°, 10.435°, 11.853°, 16.392°, 16.927°, 17.499°, 18.248°, 19.487° , 19.958°, 20.317°, 22.091°, 23.558°, 24.498°, 25.207°, 26.215°, 27.851°, 28.506°, 29.667°, 34.121°, 37.360°.
29. The crystal form D according to claim 28, its X-ray powder diffraction pattern is shown in Figure 10.
30. The crystal form D according to any one of claims 26 to 29, wherein the differential scanning calorimetry curve has an onset of an endothermic peak at 171.38±3°C.
31. Form D according to claim 30, its DSC spectrum is shown in Figure 11.
32. The crystal form D according to any one of claims 26 to 29, whose thermogravimetric analysis curve has a weight loss of 0.7461% at 99.76±3°C, and a weight loss of 5.3141% at 169.75±3°C.
33. The crystal form D according to claim 32, its TGA spectrum is shown in Figure 12.
34. Use of the crystal form according to any one of claims 1 to 33 in the preparation of a drug for treating cancer.
35. The use according to claim 34, wherein the cancer comprises lung cancer, lymphoma, esophageal cancer, ovarian cancer, pancreatic cancer, rectal cancer, brain glioma, cervical cancer, urothelial cancer, gastric cancer, intrauterine cancer Membranous cancer, liver cancer, bile duct cancer, breast cancer, colon cancer, leukemia and melanoma.

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