Crystalline form of KRAS G12D inhibitor and preparation method thereof

Abstract

The invention relates to a crystal form of a KRAS G12D inhibitor and a preparation method of the crystal form. Specifically, the invention provides A, B, C, D, E, F, G, H, I, J, K, L, M, N and O crystal forms of a compound as shown in a formula (I) and a preparation method thereof. # imgabs0 #

Description

A crystalline form of a KRAS G12D inhibitor and a preparation method thereof

[0001] This application claims the benefit of Chinese Patent Application No. 2022116420036, filed December 20, 2022. This application incorporates the entirety of the aforementioned Chinese Patent Application. Technical Field

[0002] The present disclosure belongs to the field of medical technology and relates to a crystalline form of a KRAS G12D inhibitor and a preparation method thereof. Background Art

[0003] RAS is one of the most frequently mutated oncogenes in tumors, with approximately 30% of human malignancies linked to RAS gene mutations. The RAS family includes KRAS, NRAS, and HRAS, with KRAS mutations being the most common, accounting for approximately 85%. KRAS mutations are common in solid tumors, with high frequencies found in the three most lethal human cancers: lung cancer (17%), colorectal cancer (33%), and pancreatic cancer (61%). Of the KRAS gene mutations, 97% involve mutations at amino acid residues 12 or 13, with G12D being a key mutation. Analysis of data from European and American populations shows that G12D mutations occur in 36%, 12%, and 4% of patients with pancreatic cancer, colorectal cancer, and non-small cell lung cancer, respectively.

[0004] Once activated, KRAS regulates multiple aspects of cell proliferation, survival, migration, and metabolism through numerous downstream signaling pathways, including RAF-MEK-ERK, PI3K-AKT-mTOR, and TIAM1-RAc. KRAS mutations cause the protein to remain in an activated state, leading to persistent activation of downstream signaling pathways and promoting tumorigenesis.

[0005] Because the KRAS protein lacks traditional small molecule binding sites on its surface and has an extremely high affinity for guanylate, making it extremely difficult to inhibit, it has long been considered an undruggable drug target. However, given the importance and prevalence of abnormal KRAS activation in cancer progression, KRAS has been and remains a target of great interest in drug development. Currently, with the exception of KRAS G12C inhibitors, there is still a lack of KRAS inhibitors that are effective against other mutations, leaving most patients with KRAS mutations without treatment. G12D, as a mutant that is widely and highly expressed in various tumors, has important clinical significance for the development of inhibitors against it.

[0006] Patent application PCT / CN2022 / 100016 provides a new KRAS G12D inhibitor (Formula I) with good pharmaceutical activity. To meet the needs of drug development, it is necessary to study its crystal form. The crystal form prepared in the present disclosure has good physicochemical stability, meeting the needs of drug development and storage.

[0007] Summary of the Invention

[0008] The present disclosure provides a crystalline form of a compound represented by formula (I) and a preparation method thereof.

[0009] The present disclosure provides a crystalline form A of the compound represented by formula (I), whose X-ray powder diffraction pattern has characteristic peaks at 2θ angles of 9.2, 13.1, 14.5, 16.6, 17.7, and 21.5.

[0010] In some embodiments, the crystal form A of the compound represented by formula (I) has an X-ray powder diffraction pattern with characteristic peaks at 2θ angles of 6.6, 9.2, 10.1, 10.7, 13.1, 14.5, 15.1, 16.6, 17.7, 19.6, 20.9, 21.5, 22.2, 24.0, 24.5, 24.9, 25.9, 27.2, 28.6, and 30.2.

[0011] In some embodiments, the X-ray powder diffraction pattern of Form A of the compound represented by formula (I) is shown in FIG1 .

[0012] In some embodiments, the error range of the 2θ angle of the crystal form A of the compound represented by formula (I) is ±0.2.

[0013] The present disclosure further provides a method for preparing a crystalline form of compound A represented by formula (I), comprising: mixing the compound represented by formula (I) with an appropriate amount of solvent, cooling and crystallizing; the solvent is selected from one or more of methanol and water / methanol.

[0014] The present disclosure also provides a crystal form B of the compound represented by formula (I), whose X-ray powder diffraction pattern has characteristic peaks at 2θ angles of 8.8, 12.7, 14.6, 14.9, 16.2, 18.0, and 21.4.

[0015] In some embodiments, the B form of the compound represented by formula (I) has an X-ray powder diffraction pattern with characteristic peaks at 2θ angles of 7.3, 8.8, 10.1, 10.6, 12.7, 14.6, 14.9, 16.2, 16.6, 17.3, 18.0, 19.2, 20.0, 21.4, 22.2, 24.4, 25.0, 25.7, 28.7, and 29.4.

[0016] In some embodiments, the X-ray powder diffraction pattern of Form B of the compound represented by formula (I) is shown in FIG2 .

[0017] In some embodiments, the error range of the 2θ angle of the Form B of the compound represented by formula (I) is ±0.2.

[0018] The present disclosure further provides a method for preparing a crystalline form of compound B represented by formula (I), comprising: mixing the compound represented by formula (I) with an appropriate amount of solvent, cooling and crystallizing; the solvent is selected from one or more of ethanol, isopropanol, n-propanol, ethyl acetate / ethanol, and ethyl acetate / n-heptane.

[0019] The present disclosure also provides Form C of the compound represented by formula (I), whose X-ray powder diffraction pattern has characteristic peaks at 2θ angles of 5.7, 7.6, 11.1, 11.4, 17.9, and 19.3.

[0020] In some embodiments, the X-ray powder diffraction pattern of Form C of the compound represented by Formula (I) has characteristic peaks at 2θ angles of 5.7, 6.4, 6.9, 7.6, 11.1, 11.4, 12.7, 13.9, 15.9, 17.2, 17.9, 19.3, 23.9, 24.4, 25.7, and 26.4.

[0021] In some embodiments, the X-ray powder diffraction pattern of Form C of the compound represented by formula (I) is shown in FIG3 .

[0022] In some embodiments, the error range of the 2θ angle of the Form C of the compound represented by formula (I) is ±0.2.

[0023] The present disclosure further provides a method for preparing a crystalline form of compound C represented by formula (I), comprising: mixing the compound represented by formula (I) with an appropriate amount of solvent, and crystallizing; the solvent is acetonitrile.

[0024] The present disclosure also provides a crystal form D of the compound represented by formula (I), whose X-ray powder diffraction pattern has characteristic peaks at 2θ angles of 5.8, 6.7, and 8.2.

[0025] In some embodiments, the X-ray powder diffraction pattern of the D crystal form of the compound represented by formula (I) is shown in FIG4 .

[0026] In some embodiments, the error range of the 2θ angle of the D crystal form of the compound represented by formula (I) is ±0.2.

[0027] The present disclosure further provides a method for preparing a crystalline form of compound D represented by formula (I), comprising: mixing the compound represented by formula (I) with an appropriate amount of solvent I, adding solvent II, and stirring to crystallize; the solvent I is selected from one or more of ethanol and acetonitrile / methanol, and the solvent II is selected from one or more of isopropyl acetate and methyl tert-butyl ether.

[0028] The present disclosure also provides a crystalline form E of the compound represented by formula (I), which has an X-ray powder diffraction pattern with characteristic peaks at 2θ angles of 6.7, 7.9, 9.4, 11.8, 12.3, 13.5, 14.0, 16.2, 17.5, 18.1, 20.3, 21.5, 23.6, and 24.9.

[0029] In some embodiments, the X-ray powder diffraction pattern of the E crystal form of the compound represented by formula (I) is shown in Figure 5.

[0030] In some embodiments, the error range of the 2θ angle of the E crystal form of the compound represented by formula (I) is ±0.2.

[0031] The present disclosure further provides a method for preparing a crystalline form of compound E represented by formula (I), comprising: mixing the compound represented by formula (I) with an appropriate amount of solvent, and volatilizing and crystallizing; the solvent is ethyl acetate.

[0032] The present disclosure also provides Form F of the compound represented by formula (I), which has an X-ray powder diffraction pattern with characteristic peaks at 2θ angles of 5.7, 7.8, 10.4, 11.3, 12.4, 13.7, 15.6, 16.0, 17.1, 18.0, 19.0, 19.7, 23.3, 24.2, 25.0, 25.8, and 26.9.

[0033] In some embodiments, the X-ray powder diffraction pattern of Form F of the compound represented by formula (I) is shown in FIG6 .

[0034] In some embodiments, the error range of the 2θ angle of the F crystal form of the compound represented by formula (I) is ±0.2.

[0035] The present disclosure further provides a method for preparing the crystalline form F of the compound represented by formula (I), comprising: mixing the compound represented by formula (I) with an appropriate amount of solvent and crystallizing; the solvent is methyl tert-butyl ether.

[0036] The present disclosure also provides a G crystal form of the compound represented by formula (I), whose X-ray powder diffraction pattern has characteristic peaks at 2θ angles of 7.7, 10.9, 12.8, 15.4, 20.5, and 22.0.

[0037] In some embodiments, the X-ray powder diffraction pattern of Form G of the compound represented by formula (I) is shown in FIG7 .

[0038] In some embodiments, the error range of the 2θ angle of the G crystal form of the compound represented by formula (I) is ±0.2.

[0039] The present disclosure further provides a method for preparing the crystalline form G of compound represented by formula (I), comprising: mixing the compound represented by formula (I) with an appropriate amount of solvent, stirring at room temperature for two days, and crystallizing; the solvent is dichloromethane.

[0040] The present disclosure also provides a crystalline form H of the compound represented by formula (I), which has an X-ray powder diffraction pattern with characteristic peaks at 2θ angles of 7.5, 8.6, 9.4, 12.3, 14.0, 15.9, 17.4, 18.4, 19.4, 20.5, 21.6, 22.9, 24.8, 26.6, and 27.2.

[0041] In some embodiments, the H crystal form of the compound represented by formula (I) has an X-ray powder diffraction pattern as shown in FIG8 .

[0042] In some embodiments, the error range of the 2θ angle of the H crystal form of the compound represented by formula (I) is ±0.2.

[0043] The present disclosure further provides a method for preparing the crystal form H of the compound represented by formula (I), comprising: heating the crystal form E of the compound represented by formula (I).

[0044] The present disclosure also provides Form I of the compound represented by formula (I), whose X-ray powder diffraction pattern has characteristic peaks at 2θ angles of 6.9, 15.3, 16.6, and 20.1.

[0045] In some embodiments, the Form I of the compound represented by Formula (I) has an X-ray powder diffraction pattern with characteristic peaks at 2θ angles of 5.6, 6.9, 8.3, 9.6, 11.4, 13.9, 15.3, 16.6, 18.4, 20.1, 20.6, 21.1, 22.1, 23.1, 24.5, 26.0, and 33.0.

[0046] In some embodiments, the X-ray powder diffraction pattern of Form I of the compound represented by formula (I) is shown in FIG9 .

[0047] In some embodiments, the error range of the 2θ angle of the Form I of the compound represented by formula (I) is ±0.2.

[0048] The present disclosure further provides a method for preparing a crystalline form of compound I represented by formula (I), comprising: mixing the compound represented by formula (I) with an appropriate amount of solvent, and stirring at room temperature for crystallization; the solvent is selected from one or more of methanol, ethanol, n-propanol, water / methanol, water / ethanol, water / isopropanol, acetonitrile / methanol, acetone, water / acetone, 2-butanone, acetonitrile, ethyl acetate, isopropyl acetate, n-heptane, tetrahydrofuran / ethanol, ethyl acetate / ethanol, ethyl acetate / n-heptane, isopropyl ether, and methyl tert-butyl ether.

[0049] The present disclosure also provides Form J of the compound represented by formula (I), whose X-ray powder diffraction pattern has characteristic peaks at 2θ angles of 7.2, 7.7, 8.5, 9.8, 10.6, and 14.9.

[0050] In some embodiments, the J form of the compound represented by formula (I) has an X-ray powder diffraction pattern with characteristic peaks at 2θ angles of 7.2, 7.7, 8.5, 9.8, 10.6, 12.2, 13.5, 14.9, 15.7, 17.1, 17.9, 19.1, 20.0, 20.5, 21.6, 23.3, 24.3, 25.1, 25.7, and 28.3.

[0051] In some embodiments, the X-ray powder diffraction pattern of Form J of the compound represented by formula (I) is shown in FIG10 .

[0052] In some embodiments, the error range of the 2θ angle of the J crystal form of the compound represented by formula (I) is ±0.2.

[0053] The present disclosure further provides a method for preparing the crystalline form J of the compound represented by formula (I), comprising: mixing the compound represented by formula (I) with an appropriate amount of solvent, and stirring at room temperature overnight for crystallization; the solvent is dichloromethane.

[0054] The present disclosure also provides a K crystal form of the compound represented by formula (I), whose X-ray powder diffraction pattern has characteristic peaks at 2θ angles of 5.3, 6.7, 10.7, 12.3, 13.5, 14.8, 18.1, 20.6, 21.3, and 27.3.

[0055] In some embodiments, the K crystal form of the compound represented by formula (I) has an X-ray powder diffraction pattern as shown in FIG11 .

[0056] In some embodiments, the error range of the 2θ angle of the K crystal form of the compound represented by formula (I) is ±0.2.

[0057] The present disclosure further provides a method for preparing the crystal form K of the compound represented by formula (I), comprising: mixing the compound represented by formula (I) with an appropriate amount of solvent, and crystallizing; the solvent is tetrahydrofuran.

[0058] The present disclosure also provides an L-crystalline form of the compound represented by formula (I), whose X-ray powder diffraction pattern has characteristic peaks at 2θ angles of 6.8, 8.4, 9.5, 12.3, 14.0, 16.8, 18.3, 20.3, 21.7, 22.7, and 24.8.

[0059] In some embodiments, the L-type crystal of the compound represented by formula (I) has an X-ray powder diffraction pattern as shown in FIG12 .

[0060] In some embodiments, the error range of the 2θ angle of the L crystal form of the compound represented by formula (I) is ±0.2.

[0061] The present disclosure further provides a method for preparing the L crystal form of the compound represented by formula (I), comprising: mixing the compound represented by formula (I) with an appropriate amount of solvent and crystallizing; the solvent is ethyl acetate.

[0062] The present disclosure also provides a crystalline form M of the compound represented by formula (I), which has an X-ray powder diffraction pattern having characteristic peaks at 2θ angles of 6.7, 7.8, 9.5, 11.9, 12.3, 13.0, 13.6, 14.3, 14.9, 15.8, 16.8, 17.7, 18.3, 19.1, 20.4, 21.7, 22.4, 23.9, 24.9, 26.7, 27.4, and 27.9.

[0063] In some embodiments, the X-ray powder diffraction pattern of the M crystal form of the compound represented by formula (I) is shown in Figure 13.

[0064] In some embodiments, the error range of the 2θ angle of the M crystal form of the compound represented by formula (I) is ±0.2.

[0065] The present disclosure further provides a method for preparing the crystalline form M of the compound represented by formula (I), comprising: mixing the compound represented by formula (I) with an appropriate amount of solvent and crystallizing; the solvent is isopropyl acetate.

[0066] The present disclosure also provides a crystalline form N of the compound represented by formula (I), which has an X-ray powder diffraction pattern with characteristic peaks at 2θ angles of 6.6, 7.7, 8.2, 9.1, 10.1, 11.3, 11.8, 12.6, 14.0, 15.4, 17.8, 19.0, 19.9, 21.7, 23.1, 24.0, and 25.6.

[0067] In some embodiments, the X-ray powder diffraction pattern of the N crystal form of the compound represented by formula (I) is shown in Figure 14.

[0068] In some embodiments, the error range of the 2θ angle of the N crystal form of the compound represented by formula (I) is ±0.2.

[0069] The present disclosure further provides a method for preparing crystal form N of the compound represented by formula (I), comprising: mixing the compound represented by formula (I) with an appropriate amount of solvent, and crystallizing; the solvent is acetone.

[0070] The present disclosure also provides a crystalline form O of the compound represented by formula (I), which has an X-ray powder diffraction pattern having characteristic peaks at 2θ angles of 7.0, 7.5, 8.9, 9.7, 10.5, 12.0, 12.8, 13.2, 14.0, 14.4, 15.1, 17.5, 18.1, 18.8, 19.6, 20.6, 21.2, 22.3, 24.2, 25.0, 25.9, 27.7, and 28.3.

[0071] In some embodiments, the O crystal form of the compound represented by formula (I) has an X-ray powder diffraction pattern as shown in FIG12 .

[0072] In some embodiments, the error range of the 2θ angle of the O crystal form of the compound represented by formula (I) is ±0.2.

[0073] The present disclosure further provides a method for preparing the crystal form O of the compound represented by formula (I), comprising: mixing the compound represented by formula (I) with an appropriate amount of solvent, and stirring at room temperature for crystallization; the solvent is isopropanol.

[0074] The structure and crystal form of the crystal obtained in the present invention were determined by X-ray powder diffraction (XRPD) and differential scanning calorimetry (DSC).

[0075] The crystallization method of the crystal form disclosed herein is conventional, such as evaporation crystallization, cooling crystallization or crystallization at room temperature.

[0076] The starting material used in the method for preparing the salt or crystal form disclosed herein can be any form of the compound represented by formula (I), including but not limited to: amorphous form, any crystal form, hydrate, solvate, etc.

[0077] The present disclosure further provides a pharmaceutical composition comprising the following components: (a) a crystalline form of the compound represented by formula (I); and (b) a pharmaceutically acceptable carrier, diluent, or excipient.

[0078] The present disclosure also provides a method for preparing a pharmaceutical composition, comprising the steps of mixing: (a) a crystalline form of a compound represented by formula (I); and (b) a pharmaceutically acceptable carrier, diluent, or excipient.

[0079] The present disclosure further provides a crystal form of the compound represented by the aforementioned formula (I), or use of the aforementioned composition in the preparation of a medicament for inhibiting KRAS G12D.

[0080] The present disclosure further provides the use of the crystalline form of the compound represented by the aforementioned formula (I), or the aforementioned composition in the preparation of a medicament for treating and / or preventing a disease or condition, wherein the disease or condition is cancer.

[0081] In some embodiments, the disease or condition is selected from brain cancer, thyroid cancer, head and neck cancer, nasopharyngeal cancer, pharyngeal cancer, oral cancer, salivary gland cancer, esophageal cancer, stomach cancer, lung cancer, liver cancer, kidney cancer, pancreatic cancer, gallbladder cancer, bile duct cancer, colorectal cancer, small intestine cancer, gastrointestinal stromal tumor, urothelial cancer, urethral cancer, bladder cancer, breast cancer, vaginal cancer, ovarian cancer, endometrial cancer, cervical cancer, fallopian tube cancer, testicular cancer, prostate cancer, hemangioma, leukemia, lymphoma, myeloma, skin cancer, lipoma, bone cancer, soft tissue sarcoma, neurofibroma, glioma, neuroblastoma, and glioblastoma.

[0082] In some embodiments, the disease or condition is selected from pancreatic cancer, colorectal cancer, and non-small cell lung cancer.

[0083] In the specification and claims of the application, unless otherwise indicated, the scientific and technical terms used herein have the meaning commonly understood by those skilled in the art. However, in order to better understand the disclosure, the definition and explanation of some related terms are provided below. In addition, when the definition and explanation of the term provided in the application are inconsistent with the meaning commonly understood by those skilled in the art, the definition and explanation of the term provided in the application shall prevail.

[0084] The "X-ray powder diffraction pattern or XRPD" described in the present disclosure refers to the Bragg formula 2d s(I)nθ=nλ (wherein λ is the wavelength of the X-ray, the diffraction order n is any positive integer, generally the first-order diffraction peak is taken, n=1), when the X-ray is incident on an atomic plane with a lattice plane spacing d of a crystal or a partial crystal sample at a grazing angle θ (the complementary angle of the incident angle, also known as the Bragg angle), the Bragg equation is satisfied, thereby measuring this set of X-ray powder diffraction patterns.

[0085] The "X-ray powder diffraction pattern or XRPD" referred to in the present disclosure is a pattern obtained by using Cu-Kα radiation in an X-ray powder diffractometer.

[0086] The "differential scanning calorimetry or DSC" mentioned in the present disclosure refers to measuring the temperature difference and heat flow difference between a sample and a reference object during the process of heating or maintaining a constant temperature of the sample to characterize all physical and chemical changes related to thermal effects and obtain phase change information of the sample.

[0087] The "thermogravimetric analysis or TGA" described in the present disclosure refers to the continuous measurement of the change in the mass of a sample as a function of temperature or time under programmed temperature.

[0088] The “2θ or 2θ angle” mentioned in the present disclosure refers to the diffraction angle, θ is the Bragg angle, the unit is ° or degree, and the error range of 2θ is ±0.3 or ±0.2 or ±0.1.

[0089] The "interplanar spacing or interplanar spacing (d value)" mentioned in the present disclosure refers to the selection of three non-parallel unit vectors a, b, and c connecting two adjacent lattice points in the space lattice. They divide the lattice into juxtaposed parallelepiped units, which are called interplanar spacing. The space lattice is divided according to the determined parallelepiped unit connection lines to obtain a set of straight line grids, which are called space grids or lattices. The lattice and lattice respectively use geometric points and lines to reflect the periodicity of the crystal structure. Different crystal planes have different interplanar spacings (i.e., the distance between two adjacent parallel crystal planes); the unit is Or angstrom. BRIEF DESCRIPTION OF THE DRAWINGS

[0090] FIG1 is an XRPD pattern of the crystalline form of compound A represented by formula (I);

[0091] FIG2 is an XRPD pattern of the crystalline form of compound B represented by formula (I);

[0092] FIG3 is an XRPD pattern of the crystalline form of compound C represented by formula (I);

[0093] FIG4 is an XRPD pattern of the crystalline form of compound D represented by formula (I);

[0094] FIG5 is an XRPD pattern of the crystalline form of compound E represented by formula (I);

[0095] FIG6 is an XRPD pattern of the crystalline form F of compound represented by formula (I);

[0096] FIG7 is an XRPD pattern of the crystalline form G of compound represented by formula (I);

[0097] FIG8 is an XRPD pattern of Form H of the compound represented by formula (I);

[0098] FIG9 is an XRPD pattern of the crystalline form of compound I represented by formula (I);

[0099] FIG10 is an XRPD pattern of the crystalline form J of the compound represented by formula (I);

[0100] FIG11 is an XRPD pattern of the crystal form K of compound represented by formula (I);

[0101] FIG12 is an XRPD pattern of the crystalline form L of the compound represented by formula (I);

[0102] FIG13 is an XRPD pattern of the crystalline form M of the compound represented by formula (I);

[0103] FIG14 is an XRPD pattern of Form N of the compound represented by formula (I);

[0104] FIG15 is an XRPD pattern of Form O of the compound represented by formula (I);

[0105] FIG16 is an XRPD pattern of the amorphous form of the compound represented by formula (I). DETAILED DESCRIPTION

[0106] The present disclosure will be explained in more detail below with reference to embodiments. The embodiments of the present disclosure are only used to illustrate the technical solutions of the present disclosure and are not intended to limit the essence and scope of the present disclosure.

[0107] The structures of the compounds were determined by nuclear magnetic resonance (NMR) and / or mass spectrometry (MS). -6 The unit of ppm is given. NMR measurements were performed using a Bruker AVANCE-400 NMR spectrometer or a Bruker AVANCE NEO 500M NMR spectrometer. The solvents used for the measurements were deuterated dimethyl sulfoxide (DMSO-d6), deuterated chloroform (CDCl3), or deuterated methanol (CD3OD), and tetramethylsilane (TMS) was used as the internal standard.

[0108] MS was measured using an Agilent 1200 / 1290 DAD-6110 / 6120 Quadrupole MS liquid spectrometer-mass spectrometer (manufacturer: Agilent, MS model: 6110 / 6120 Quadrupole MS).

[0109] waters ACQuity UPLC-QD / SQD (Manufacturer: waters, MS model: waters ACQuity Qda Detector / waters SQ Detector)

[0110] THERMO Ultimate 3000-Q Exactive (Manufacturer: THERMO, MS model: THERMO Q Exactive)

[0111] High performance liquid chromatography (HPLC) analysis was performed using Agilent HPLC 1200DAD, Agilent HPLC 1200VWD and Waters HPLC e2695-2489 high pressure liquid chromatographs.

[0112] Chiral HPLC analysis was performed using an Agilent 1260 DAD high performance liquid chromatograph.

[0113] High performance liquid chromatography (HPLC) was performed using Waters 2545-2767, Waters 2767-SQ Detecor2, Shimadzu LC-20AP, and Gilson GX-281 preparative chromatographs.

[0114] Chiral preparations were performed using a Shimadzu LC-20AP preparative chromatograph.

[0115] The CombiFlash rapid preparation instrument used was Combiflash Rf200 (TELEDYNE ISCO).

[0116] The thin layer chromatography silica gel plate uses Yantai Huanghai HSGF254 or Qingdao GF254 silica gel plate. The specification of the silica gel plate used in thin layer chromatography (TLC) is 0.15mm~0.2mm, and the specification used for thin layer chromatography separation and purification products is 0.4mm~0.5mm.

[0117] Silica gel column chromatography generally uses Yantai Huanghai 200-300 mesh silica gel as the carrier.

[0118] Average kinase inhibition rate and IC 50 The values ​​were determined using a NovoStar microplate reader (BMG, Germany).

[0119] The known starting materials disclosed herein can be synthesized by methods known in the art, or can be purchased from ABCR GmbH & Co.KG, Acros Organics, Aldrich Chemical Company, Accela ChemBio Inc, Darui Chemicals, and other companies.

[0120] Unless otherwise specified in the examples, all reactions can be carried out under an argon atmosphere or a nitrogen atmosphere.

[0121] Argon atmosphere or nitrogen atmosphere means that the reaction bottle is connected to an argon or nitrogen balloon with a capacity of about 1 L.

[0122] Hydrogen atmosphere means that the reaction bottle is connected to a hydrogen balloon with a capacity of about 1L.

[0123] The pressurized hydrogenation reaction uses a Parr 3916EKX hydrogenator and a Qinglan QL-500 hydrogen generator or an HC2-SS hydrogenator.

[0124] The hydrogenation reaction is usually carried out by evacuating the chamber and filling it with hydrogen, and the operation is repeated three times.

[0125] A CEM Discover-S 908860 microwave reactor was used for the microwave reaction.

[0126] Unless otherwise specified in the examples, the solution refers to an aqueous solution.

[0127] Unless otherwise specified in the examples, the reaction temperature is room temperature, 20°C to 30°C.

[0128] The reaction progress in the examples was monitored by thin layer chromatography (TLC). The developing solvent used in the reaction, the eluent system for column chromatography used to purify the compound, and the developing solvent system for thin layer chromatography included: A: dichloromethane / methanol system, B: n-hexane / ethyl acetate. The volume ratio of the solvents was adjusted according to the polarity of the compound, and a small amount of alkaline or acidic reagents such as triethylamine and acetic acid could also be added for adjustment.

[0129] Where the compounds in the examples contain two or more chiral centres, the relative stereochemistry of these compounds was determined by NMR studies and / or X-ray diffraction. In these cases, the prefix "rel" followed by the R / S nomenclature is used to identify these compounds, where the R / S only provides relative stereochemical information and does not indicate absolute stereochemistry. For example, express A 1:1 mixture of 2,3-dimethoxy-2-nitropropene is a racemate.

[0130] XRPD is X-ray powder diffraction detection: the measurement was carried out using a BRUKER D8 X-ray diffractometer, and the specific collection information was: Cu anode (40kV, 40mA), Cu-Kα1 ray Kα2 rays Kβ rays Scanning mode: θ / 2θ, scanning range (2θ range): 3 to 45°.

[0131] DSC is differential scanning calorimetry: the measurement was performed using a METTLER TOLEDO DSC 3+ differential scanning calorimeter with a heating rate of 10°C / min. The specific temperature range was referred to the corresponding spectrum (mostly 25-300 or 25-350°C), and a nitrogen purge rate of 50 mL / min.

[0132] TGA is thermogravimetric analysis: the test was performed using a METTLER TOLEDO TGA 2 thermogravimetric analyzer with a heating rate of 10°C / min. The specific temperature range was referred to the corresponding spectrum (mostly 25-400°C), and a nitrogen purge rate of 50 mL / min.

[0133] DVS stands for dynamic moisture sorption: the test uses SMS DVS Advantage. At 25°C, the humidity changes from 50% to 95% to 0% to 95% to 50%, with a step of 10% (the last step is 5%) (the specific humidity range is based on the corresponding spectrum, and the method listed here is mostly used). The judgment standard is dm / dt no more than 0.002%.

[0134] Example 1

[0135] (±)-rel-(1R,2R,5S)-tert-butyl 2-(((tert-butyldimethylsilyl)oxy)methyl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate 1h

[0136] first step

[0137] (±)-5-methoxy-3,4-dihydro-2H-pyrrole-2-carboxylic acid methyl ester 1b

[0138] Methyl (±)-2-pyrrolidone-5-carboxylate 1a (100 g, 698.61 mmol, Shanghai Bidex) and dimethyl sulfate (110 g, 872.10 mmol) were mixed and reacted at 60°C for 16 hours. The reaction solution was cooled to room temperature and poured into a solution of triethylamine (100 g) and methyl tert-butyl ether (150 mL) under an ice bath. After extraction with methyl tert-butyl ether (300 mL × 6), the mixture was concentrated under reduced pressure to give the crude title compound 1b (90 g, yield: 81.9%), which was used directly in the next step without purification.

[0139] MS m / z (ESI): 158.1 [M+1].

[0140] Step 2

[0141] (±)-5-(2-methoxy-1-nitro-2-oxoethylidene)pyrrolidine-2-carboxylic acid methyl ester 1c

[0142] The crude compound 1b (90 g, 572.64 mmol) and methyl nitroacetate (68.18 g, 572.63 mmol) were mixed, heated to 60°C and stirred for 30 hours. After the reaction solution was cooled to room temperature, ethyl acetate (300 mL) was added, stirred for 0.5 hours, and filtered. The filter cake was dried to obtain the title compound 1c (70 g, yield: 50%), which was used directly in the next step without purification.

[0143] MS m / z (ESI): 245.1 [M+1].

[0144] Step 3

[0145] 4-Oxo-3,8-diazabicyclo[3.2.1]octane-2-carboxylic acid methyl ester (diastereomeric mixture) 1d

[0146] Crude compound 1c (14 g, 57.3 mmol) was dissolved in 600 mL of methanol, and 10% palladium-on-carbon catalyst (wet) (14 g) was added. The atmosphere was replaced with hydrogen three times, and the reaction was stirred for 48 hours. The reaction solution was filtered through celite, and the filtrate was concentrated to obtain the crude title compound 1d (10 g, 94.6% yield), which was used directly in the next step without purification.

[0147] MS m / z (ESI): 185.2 [M+1].

[0148] Step 4

[0149] 8-(tert-Butyl)-2-methyl(±)-rel-(1R,2R,5S)-4-oxo-3,8-diazabicyclo[3.2.1]octane-2,8-dicarboxylate 1e: The crude compound 1d (10 g, 54.2 mmol) was dissolved in 300 mL of dichloromethane. Triethylamine (16 g, 158.12 mmol) and di-tert-butyl dicarbonate (11 g, 50.4 mmol, Shanghai Shaoyuan) were added under ice-cooling. The reaction was stirred for 14 hours. The reaction solution was concentrated under reduced pressure and the residue was purified by silica gel column chromatography with eluent System B to give the title compound 1e (3.3 g, yield: 21.3%).

[0150] MS m / z (ESI): 285.2 [M+1].

[0151] HPLC analysis: retention time 1.02 minutes, purity: 98.5% (chromatographic column: ACQUITY C18, 1.7 μm, 2.1*50 mm; mobile phase: water (10 mM ammonium bicarbonate), acetonitrile, gradient ratio: acetonitrile 10%-95%).

[0152] Step 5

[0153] 8-(tert-Butyl)2-methyl(±)-rel-(1R,2R,5S)-3,8-diazabicyclo[3.2.1]octane-2,8-dicarboxylate 1f

[0154] Compound 1e (400 mg, 1.4 mmol) was dissolved in 2 mL of tetrahydrofuran, and 3.5 mL of a 2 M solution of borane dimethyl sulfide complex in tetrahydrofuran was added. The mixture was stirred and reacted for 14 hours. Methanol was added to the reaction solution to quench the reaction, and the reaction was continued at 50°C for 14 hours. The reaction solution was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography with eluent System A to obtain the title compound 1f (176 mg, yield: 46.2%).

[0155] MS m / z (ESI): 271.2 [M+1].

[0156] Step 6

[0157] (±)-rel-(1R, 2R, 5S)-2-(hydroxymethyl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylic acid tert-butyl ester 1g

[0158] Compound 1f (1 g, 3.69 μmol) was dissolved in 15 mL of tetrahydrofuran, and 4.4 mL of 1 M lithium aluminum hydride solution in tetrahydrofuran was added. The mixture was stirred at 0°C for 1 hour. 0.2 mL of water, 0.2 mL of 15% aqueous sodium hydroxide solution, and 0.4 mL of water were added to the reaction solution in sequence. Anhydrous sodium sulfate was then added and stirred for 10 minutes. The mixture was filtered and the filtrate was concentrated to give 1 g of the title compound (430 mg, yield: 47.9%). The product was used directly in the next step without purification.

[0159] MS m / z (ESI): 243.1 [M+1].

[0160] Step 7

[0161] (±)-rel-(1R,2R,5S)-tert-butyl 2-(((tert-butyldimethylsilyl)oxy)methyl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate 1h

[0162] 1 g (8.8 g, 36.3 mmol), tert-butyldimethylsilyl chloride (16 g, 106.1558 mmol), and 4-dimethylaminopyridine (4 g, 32.4739 mmol) were dissolved in 200 mL of dichloromethane, and triethylamine (15 g, 148.23 mmol, 21.4286 mL) was added. The reaction mixture was stirred for 16 hours, and the reaction solution was concentrated under reduced pressure. The residue was purified by silica gel column chromatography with eluent System B to give the title compound 1h (8 g, yield: 61.7%).

[0163] MS m / z (ESI): 357.1 [M+1].

[0164] Example 2

[0165] 5-Ethyl-6-fluoro-4-((5aS,6S,9R)-1-fluoro-12-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-5a,6,7,8,9,10-hexahydro-5H-4-oxa-3,10a,11,13,14-pentaaza-6,9-methano[1,8-ab]heptylcyclo[2-yl]naphthol-2-ol 1-p1

[0166] Step 8

[0167] 2,6-Dichloro-3-fluoropyridin-4-amine 1j

[0168] Compound 4-amino-2,6-dichloropyridine 1i (5 g, 30.6 mmol, Shanghai Bidex) was dissolved in 20 mL of N,N-dimethylformamide and 20 mL of acetonitrile, and 1-chloromethyl-4-fluoro-1,4-diazabicyclo[2.2.2]octane di(tetrafluoroborate) salt (13 g, 36.8 mmol) was added. The mixture was reacted at 80°C for 0.5 hour. The reaction solution was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography with eluent System B to give the title compound 1j (2.2 g, yield: 39.6%).

[0169] Step 9

[0170] tert-Butyl 4-((tert-Butoxycarbonyl)amino)-2,6-dichloro-5-fluoronicotinate 1k

[0171] Compound 1j (1.8 g, 9.94 mmol) was dissolved in tetrahydrofuran (50 mL). 20 mL of a 2 M solution of sodium bistrimethylsilylamide in tetrahydrofuran was added under ice-cooling. The mixture was stirred for 0.5 hour, and then di-tert-butyl dicarbonate (6.5 g, 29.7 mmol) was added. The mixture was stirred for 14 hours. The reaction mixture was quenched by the addition of saturated aqueous ammonium chloride and extracted with ethyl acetate (50 mL x 3). The organic phases were combined and dried over anhydrous sodium sulfate. The desiccant was removed by filtration, and the filtrate was concentrated under reduced pressure. The residue was purified with eluent System B to give the title compound 1k (1 g, yield: 26.3%), which was used in the next step without further purification.

[0172] MS m / z (ESI): 381.1 [M+1].

[0173] Step 10

[0174] tert-Butyl 4-amino-2,6-dichloro-5-fluoronicotinate

[0175] Compound 1k (1 g, 2.62 mmol) was dissolved in ethyl acetate (8 mL), and 3 mL of 4 M hydrochloric acid in dioxane was added. The reaction was stirred for 2 hours. The pH was adjusted to neutral with 4 M aqueous sodium hydroxide solution under ice bath, and extracted with ethyl acetate (10 mL x 3). The organic phases were combined and dried over anhydrous sodium sulfate. The desiccant was removed by filtration, and the filtrate was concentrated under reduced pressure. The residue was purified with eluent system B to give the crude title compound 1l (500 mg, yield: 67.8%).

[0176] MS m / z (ESI): 281.1 [M+1].

[0177] Step 11

[0178] 2,6-dichloro-5-fluoro-4-(3-(2,2,2-trichloroacetyl)ureido)nicotinate tert-butyl ester 1m

[0179] The crude compound 1l (500 mg, 1.77 mmol) was dissolved in tetrahydrofuran (10 mL), and trichloroacetyl isocyanate (670 mg, 3.55 mmol) was added. The mixture was stirred for 30 minutes, and the reaction solution was concentrated under reduced pressure to obtain the crude title compound 1m (835 mg, yield: 99.7%). The product was used directly in the next step without purification.

[0180] MS m / z (ESI): 467.9 [M+1].

[0181] Step 12

[0182] 5,7-Dichloro-8-fluoro-pyrido[4,3-d]pyrimidine-2,4-diol 1n

[0183] The crude compound 1m (835 mg, 1.77 mmol) was dissolved in 7 M ammonia methanol solution (10 mL) and stirred for 1 hour. The reaction solution was concentrated under reduced pressure. Methyl tert-butyl ether (10 mL) was added to the residue, stirred for 0.5 hour, and then filtered. The filter cake was dried to give the crude title compound 1n (400 mg, yield: 89.9%), which was used directly in the next step without purification.

[0184] MS m / z (ESI): 249.9 [M+1].

[0185] Step 13

[0186] 2,4,5,7-Tetrachloro-8-fluoro-pyrido[4,3-d]pyrimidine 1o

[0187] The crude compound 1n (300 mg, 1.19 mmol) was dissolved in phosphorus oxychloride (6 mL), and N,N-diisopropylethylamine (800 mg, 6.19 mmol) was added. The reaction was stirred at 110°C for 3 hours. The reaction solution was cooled to room temperature and concentrated under reduced pressure to obtain the crude title compound 1o (344 mg, yield: 97.7%). The product was used directly in the next step without purification.

[0188] MS m / z (ESI): 285.8 [M+1].

[0189] Step 14

[0190] (±)-rel-(1R,2R,5S)-2-(((tert-butyldimethylsilyl)oxy)methyl)-3-(2,5,7-trichloro-8-fluoropyrido[4,3-d]pyrimidin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylic acid tert-butyl ester 1p

[0191] Compound 1o (1.0 g, 3.48 mmol) and N,N-diisopropylethylamine (0.9 g, 6.9 mmol) were dissolved in 15 mL of dichloromethane, and 1h (1.25 g, 3.5 mmol) was added at -78°C. The temperature was maintained with stirring for 1 hour, and then the reaction was returned to room temperature for 16 hours. The reaction solution was concentrated under reduced pressure, and the residue was purified with eluent system B to obtain the crude title compound 1p (1.56 g, yield: 73.7%).

[0192] MS m / z (ESI): 606.2 [M+1].

[0193] Step 15

[0194] tert-Butyl (1S,2S,5R)-2-(((tert-butyldimethylsilyl)oxy)methyl)-3-(5,7-dichloro-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)pyrido[4,3-d]pyrimidin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate 1q-1 and

[0195] Diastereomeric mixture of tert-butyl (1R,2R,5S)-2-(((tert-butyldimethylsilyl)oxy)methyl)-3-(5,7-dichloro-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)pyrido[4,3-d]pyrimidin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate 1q-2

[0196] Compound 1p (1.4 g, 2.3 mmol) was dissolved in 1,4-dioxane (20 mL), and ((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methanol (650 mg, 4.08 mmol, Yao Ming), N,N-diisopropylethylamine (1.5 g, 11.6 mmol), and 4A molecular sieves (1.4 g) were added. The mixture was stirred at 105° C. for 6 hours. The reaction solution was cooled to room temperature, filtered, and concentrated under reduced pressure to give a crude diastereomeric mixture of the title compounds 1q-1 and 1q-2 (1.68 g, yield: 99.8%). The product was used directly in the next step without purification.

[0197] MS m / z (ESI): 729.2 [M+1].

[0198] Step 16

[0199] (5aS,6S,9R)-2-chloro-1-fluoro-12-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-5a,6,7,8,9,10-hexahydro-5H-4-oxa-3,10a,11,13,14-pentaaza-6,9-methano[1,8-ab]heptyl-14-carboxylate tert-butyl ester 1r-1

[0200] and a diastereomeric mixture of (5aR,6R,9S)-2-chloro-1-fluoro-12-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-5a,6,7,8,9,10-hexahydro-5H-4-oxa-3,10a,11,13,14-pentaaza-6,9-methano[1,8-ab]heptyl-14-carboxylic acid tert-butyl ester 1r-2

[0201] Tetrabutylammonium fluoride (2.59 g, 11.51 mmol) was added to the diastereomeric mixture of the crude products 1q-1 and 1q-2 (1.68 g, 2.3 mmol) and stirred at room temperature for 16 hours; the reaction solution was concentrated under reduced pressure, and the residue was purified with eluent System B to give the diastereomeric mixture of the title compounds 1r-1 and 1r-2 (1.0 g, yield: 75.0%).

[0202] MS m / z (ESI): 579.2 [M+1].

[0203] Step 17

[0204] (5aS,6S,9R)-2-(8-ethyl-7-fluoro-3-(methoxymethoxy)naphthalen-1-yl)-1-fluoro-12-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5a,6,7,8,9,10-hexahydro-5H-4-oxa-3,10a,11,13,14-pentaaza-6,9-methano[1,8-ab]heptyl-14-carboxylic acid tert-butyl ester 1s-1 and

[0205] A 1:1 mixture of diastereomers of (5aR,6R,9S)-2-(8-ethyl-7-fluoro-3-(methoxymethoxy)naphthalen-1-yl)-1-fluoro-12-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-5a,6,7,8,9,10-hexahydro-5H-4-oxa-3,10a,11,13,14-pentaaza-6,9-methano[1,8-ab]heptane-14-carboxylic acid tert-butyl ester 1s-2

[0206] A diastereomeric mixture of compounds 1r-1 and 1r-2 (300 mg, 518.1 μmol), 2-(8-ethyl-7-fluoro-3-(methoxymethoxy)naphthalen-1-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (280 mg, 777.2 μmol, prepared by the method disclosed in Intermediate 18 on page 104 of the specification of patent application "WO2021 / 041671"), tetrakis(triphenylphosphine)palladium (120 mg, 103.8 μmol), and cesium carbonate (506 mg, 1.55 mmol) were dissolved in 6 mL of a mixed solution of 1,4-dioxane and water (V:V=5:1). Under nitrogen atmosphere, the reaction was carried out at 100°C for 6 hours. The reaction solution was concentrated under reduced pressure to give a crude product (400 mg) of a 1:1 mixture of diastereomers of the title compound 1s-1 and 1s-2. The product was used directly in the next reaction without purification.

[0207] MS m / z (ESI): 777.2 [M+1].

[0208] Step 18

[0209] 5-ethyl-6-fluoro-4-((5aS,6S,9R)-1-fluoro-12-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-5a,6,7,8,9,10-hexahydro-5H-4-oxa-3,10a,11,13,14-pentaaza-6,9-methano[1,8-ab]heptacyclo-2-yl)naphthalen-2-ol 1-p1 and

[0210] 5-Ethyl-6-fluoro-4-((5aR,6R,9S)-1-fluoro-12-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-5a,6,7,8,9,10-hexahydro-5H-4-oxa-3,10a,11,13,14-pentaaza-6,9-methano[1,8-ab]heptacyclo[2-yl]naphtho ...1,8-ab]heptacyclo[1,8-ab]heptacyclo[1,8-ab]heptacyclo[1,8-ab]heptacyclo[1,8-ab]heptacyclo[1,8-ab]heptacyclo[1,8-ab]heptacyclo[1,8-ab]heptacyclo[1,8-ab]heptacyclo[1,8-ab]heptacyclo[1,8-ab]heptacyclo[1,8-ab]he

[0211] The crude diastereomeric mixture of compounds 1s-1 and 1s-2 (160 mg, 205.9 μmol) was dissolved in ethyl acetate (5 mL), and 1 mL of 4 M hydrochloric acid and dioxane solution was added. The reaction was carried out at 0°C for 2 hours. The reaction solution was concentrated under reduced pressure, and the residue was purified by high performance liquid chromatography (Waters-2545, column: SharpSil-T C18, 30*150 mm, 5 μm; mobile phase: aqueous phase (10 mmol / L ammonium bicarbonate) and acetonitrile, gradient ratio: acetonitrile 38%-45%, flow rate: 30 mL / min) to give a 1:1 mixture of diastereomers of the title compounds 1-p1 and 1-p2 (10 mg, yield: 7.2%).

[0212] MS m / z (ESI): 633.2 [M+1].

[0213] 1 H NMR (500MHz, CD3OD): δ7.67 (ddd, 1H), 7.32-7.21 (m, 2H), 7.11-7.01 (m, 1H), 5 .38-5.35(m, 2H), 5.11-5.03(m, 1H), 4.64-4.59(m, 1H), 4.52-4.46(m, 1H), 4. 34-4.29(m, 1H), 4.25(dd, 1H), 4.18-4.12(m, 1H), 3.74(br, 1H), 3.65(br, 1H) , 3.26-3.23(m, 3H), 3.05-3.01(m, 1H), 2.61-1.81(m, 12H), 0.94-0.82(m, 3H).

[0214] The diastereomeric mixture of compounds 1-p1 and 1-p2 was separated by chiral column (Shimadzu LC-20AP, chromatographic column: DAICEL 25*250mm, 10μm; mobile phase A: n-hexane, mobile phase B: ethanol (0.1% 7M NH3 in MeOH) and, gradient ratio: A:B:40:60, flow rate: 30mL / min) to give the title compounds 1-p1 (26mg, yield: 43.3%) and 1-p2 (26mg, yield: 43.3%).

[0215] Single configuration compound (shorter retention time) 1-p2: (26 mg, yield: 43.3%).

[0216] MS m / z (ESI): 633.2 [M+1].

[0217] Chiral HPLC analysis: retention time 7.89 minutes, purity: 99% (chromatographic column: DAICEL 250*4.6mm, 5μm; mobile phase: n-hexane and ethanol (containing 0.2% diethylamine), flow rate: 1.0mL / min).

[0218] 1H NMR (500MHz, CD3OD): δ7.67 (ddd, 1H), 7.32-7.21 (m, 2H), 7.11-7.01 (m, 1H), 5.38-5.27(m, 2H), 5.11-5.03(m, 1H), 4.64-4.59(m, 1H), 4.52-4.44(m, 1H), 4.33(d, 1H), 4.24(dd, 1H), 4.18-4.12(m, 1H), 3.75(br, 1H), 3.66(br, 1H), 3 .27-3.18(m, 3H), 3.05-3.03(m, 1H), 2.60-1.81(m, 12H), 0.93-0.82(m, 3H).

[0219] Single configuration compound (longer retention time) 1-p1: (26 mg, yield: 43.3%).

[0220] MS m / z (ESI): 633.2 [M+1].

[0221] Chiral HPLC analysis: retention time 13.8 minutes, purity: 99% (chromatographic column: DAICEL 250*4.6mm, 5μm; mobile phase: n-hexane and ethanol (containing 0.2% diethylamine), flow rate: 1.0mL / min).

[0222] 1 H NMR (500MHz, CD3OD): δ7.67 (ddd, 1H), 7.32-7.21 (m, 2H), 7.11-7.01 (m, 1H), 5 .38-5.35(m, 2H), 5.11-5.03(m, 1H), 4.64-4.59(m, 1H), 4.52-4.46(m, 1H), 4. 34-4.29(m, 1H), 4.25(dd, 1H), 4.18-4.12(m, 1H), 3.74(br, 1H), 3.65(br, 1H) , 3.26-3.23(m, 3H), 3.05-3.01(m, 1H), 2.61-1.81(m, 12H), 0.94-0.82m, 3H).

[0223] Compound 1-p1 is the compound represented by formula (I) in the present disclosure. X-ray powder diffraction analysis shows that the compound is amorphous. The X-ray powder diffraction spectrum is shown in FIG16 .

[0224] Biological evaluation

[0225] Test Example 1: Biological Evaluation of ERK Phosphorylation Inhibition Experiment in AGS Cells (HTRF Method)

[0226] 1. Test Purpose

[0227] This study examined the inhibitory effect of compound 1-p1 on ERK phosphorylation in cells. 50 The size of the compound was evaluated to determine its inhibitory effect on the KRAS target.

[0228] 2. Experimental Methods

[0229] AGS cells (Nanjing Kebai, CBP60476) were cultured in RPMI1640 (Hyclone, SH30809.01) complete medium supplemented with 10% fetal bovine serum. On the first day of the experiment, AGS cells were seeded at a density of 40,000 cells / well in a 96-well plate using complete medium, with 190 μL of cell suspension per well. The cells were incubated overnight at 37°C in a 5% CO2 incubator.

[0230] The next day, 10 μL of a serial dilution of the test compound prepared in complete medium was added to each well. The final concentration of the compound was a five-fold serial dilution starting from 10 μM, with nine concentration points. A blank control containing 0.5% DMSO was set up. The plate was incubated in a cell culture incubator at 37°C and 5% CO2 for 1 hour. After incubation, the 96-well cell culture plate was removed, the medium was aspirated, and 200 μL of PBS (Shanghai Yuanpei Biotechnology Co., Ltd., B320) was added to each well for washing. The PBS was aspirated, and 50 μL of lysis buffer (Cisbio, 64KL1FDF) containing blocking reagent (Cisbio, 64KB1AAC) was added to each well. The plate was shaken on a shaker at room temperature for 40 minutes. After lysis, pipette to mix thoroughly. Transfer 16 μL of lysate per well to two HTRF 96-well assay plates (Cisbio, 66PL96100). Then, add 4 μL of premixed phosphorylated ERK1 / 2 antibody solution (Cisbio, 64AERPEG) or 4 μL of premixed total ERK1 / 2 antibody solution (Cisbio, 64NRKPEG) to each plate. Seal the plates with sealing film, centrifuge in a microplate centrifuge for 1 minute, and incubate overnight at room temperature in the dark.

[0231] On the third day, the fluorescence values ​​were read using an ENVISION multifunctional microplate reader (PerkinElmer, ENVISION) with excitation at 337 nm and emission at 665 nm and 620 nm.

[0232] 3. Data Analysis and Results

[0233] Graphpad Prism software was used to calculate the IC of the inhibitory activity of the compound based on the compound concentration and the ratio of phosphorylated ERK / total ERK. 50The results showed that the IC value of compound 1-p1 50 The concentration of 1% β-catenin was 0.4 nM, which had a good inhibitory effect on ERK phosphorylation in AGS cells.

[0234] Test Example 2: Biological Evaluation of GP2d and AGS Cell 3D Proliferation Inhibition Experiment

[0235] 1. Test Purpose

[0236] The inhibitory effect of compound 1-p1 on the KRAS target was evaluated by testing the 3D proliferation inhibitory effect of compound 1-p1 on GP2d and AGS cells.

[0237] 2. Experimental Methods

[0238] GP2d cells (Nanjing Kebai, CBP60010) were cultured in complete medium (DMEM / high glucose medium (Hyclone, SH30243.01) supplemented with 10% fetal bovine serum (Corning, 35-076-CV). On the first day of the experiment, GP2d cells were seeded at a density of 1000 cells / well in a 96-well low attachment plate (Corning, CLS7007-24EA) using complete medium. 90 μL of cell suspension was added to each well, centrifuged at 2000 rpm for 5 minutes at room temperature, and then incubated overnight in a 37°C, 5% CO2 cell incubator.

[0239] AGS cells (Nanjing Kebai, CBP60476) were cultured in complete medium (RPMI1640 medium (Hyclone, SH30809.01) supplemented with 10% fetal bovine serum (Corning, 35-076-CV). On the first day of the experiment, AGS cells were seeded at a density of 1000 cells / well in a 96-well low attachment plate (Corning, CLS7007-24EA) using complete medium. 90 μL of cell suspension was added to each well, centrifuged at 2000 rpm for 5 minutes at room temperature, and then incubated overnight at 37°C in a 5% CO2 incubator.

[0240] On the second day, 10 μL of a serial dilution of the test compound prepared in complete culture medium was added to each well. The final concentration of the compound for GP2d cells was 9 concentration points of a 3-fold serial dilution starting from 1 μM, and the final concentration of the compound for AGS cells was 9 concentration points of a 3-fold serial dilution starting from 10 μM. A blank control containing 0.5% DMSO was set up for both. The well plate was placed in a cell culture incubator at 37°C and 5% CO2 for 120 hours. On the seventh day, the 96-well cell culture plate was removed and 50 μL of the test compound was added to each well. 3D reagent (Promega, G9682) was shaken at room temperature for 25 minutes, then pipetted to mix thoroughly and 50 μL was transferred to a white opaque 96-well plate (PE, 6005290). The luminescent signal was read using a multi-function microplate reader (PerkinElmer, ENVISION).

[0241] 3. Data Analysis and Results

[0242] The IC of the inhibitory activity of the compounds was calculated using Graphpad Prism software 50 The results showed that the IC 50 The IC for GP2d cells is 5.8 nM. 50 The concentration of the compound 1-p1 was 0.9 nM, and the compound 1-p1 had a good inhibitory effect on the 3D proliferation of AGS and GP2d cells.

[0243] Test Example 3: SPR method to detect the affinity of compounds to KRAS protein isoforms G12D or WT

[0244] Biotinylated Avi-KRAS-WT or Avi-KRAS-G12D was diluted to 20 μg / mL in 1× HBS-P+ (Cat.#BR1006-71) buffer containing 100 mM MgCl₂ and then passed through channel 2 of an SA (Cat.#BR1005-31) biosensor chip for 420 s, achieving a binding level of approximately 5000-7000 RU. Compound samples were then injected for 120 s, followed by a 720 s dissociation period. The assay was performed in single-cycle kinetic mode. Reaction signals were monitored in real time on a Biacore 8K instrument to generate binding and dissociation curves. Data were analyzed using Biacore 8K evaluation software, with a 1:1 model used for data fitting and affinity data. The results showed that the KD value of compound 1-p1 for KRAS G12D was 0.03E-09 mol / L, and the KD value for WT was 1.54E-09 mol / L. Compound 1-p1 has good affinity with KRAS protein isoform G12D or WT.

[0245] Example 3 Preparation of the Crystalline Form of Compound A Represented by Formula (I)

[0246] 8 mg of the compound represented by formula (I) was weighed and dissolved in 0.04 mL of methanol. The mixture was cooled to 5°C, stirred to crystallize, centrifuged, and the solid dried in vacuo to obtain the product. X-ray powder diffraction analysis revealed an XRPD pattern as shown in Figure 1 and characteristic peak positions as shown in Table 1. This product was defined as Form A of the compound represented by formula (I). The DSC spectrum showed endothermic peaks at 74.81°C, 140.13°C, and 196.78°C.

[0247] Table 1

[0248] Example 4 Preparation of the Crystalline Form of Compound A Represented by Formula (I)

[0249] 8 mg of the compound of formula (I) was weighed and dissolved in 0.04 mL of 10% water / methanol (v / v). The mixture was cooled to 5°C, stirred to crystallize, centrifuged, and the solid dried under vacuum to obtain the product. X-ray powder diffraction analysis confirmed that the product was Form A of the compound of formula (I).

[0250] Example 5 Preparation of the Crystalline Form of Compound B Represented by Formula (I)

[0251] 8 mg of the compound represented by formula (I) was added to 0.04 mL of ethanol, stirred and dissolved at room temperature, cooled to 5°C, stirred to precipitate, centrifuged, and the solid dried in vacuo to obtain the product. X-ray powder diffraction analysis revealed an XRPD spectrum as shown in Figure 2 and characteristic peak positions as shown in Table 2, defining the product as Form B of the compound represented by formula (I). The DSC spectrum showed an endothermic peak at 226.81°C.

[0252] Table 2

[0253] Example 6 Preparation of the Crystalline Form of Compound B Represented by Formula (I)

[0254] 8 mg of the compound represented by formula (I) was added to 0.8 mL of ethyl acetate / n-heptane (v / v = 1:1), stirred at room temperature for 2 days, centrifuged, and the solid dried under vacuum to obtain the product. X-ray powder diffraction analysis showed that the product was Form B of the compound represented by formula (I).

[0255] Example 7 Preparation of Crystalline Form C of Compound Represented by Formula (I)

[0256] 8 mg of the compound represented by formula (I) was dissolved in 0.8 mL of acetonitrile at 50°C, cooled to 5°C, stirred to crystallize, centrifuged, and the solid dried in vacuo to obtain the product. X-ray powder diffraction analysis revealed an XRPD pattern as shown in Figure 3 and characteristic peak positions as shown in Table 3. This product was defined as Form C of the compound represented by formula (I). The DSC spectrum showed an endothermic peak at 198.44°C.

[0257] Table 3

[0258] Example 9 Preparation of Crystalline Form D of Compound Represented by Formula (I)

[0259] 8 mg of the compound represented by formula (I) was added to 0.05 mL of ethanol and dissolved by stirring at room temperature. 0.3 mL of isopropyl acetate was added, stirred to separate crystals, centrifuged, and the solid dried under vacuum to obtain the product. X-ray powder diffraction analysis revealed an XRPD pattern as shown in Figure 4 and characteristic peak positions as shown in Table 4. This product was defined as Form D of the compound represented by formula (I). The DSC spectrum showed an endothermic peak at 178.74°C.

[0260] Table 4

[0261] Example 10 Preparation of the Crystalline Form of Compound E Represented by Formula (I)

[0262] 8 mg of the compound represented by formula (I) was dissolved in 0.6 mL of ethyl acetate and evaporated to obtain the product. X-ray powder diffraction analysis revealed an XRPD pattern as shown in Figure 5 and characteristic peak positions as shown in Table 5. This product was defined as Form E of the compound represented by formula (I). The DSC spectrum showed endothermic peaks at 44.15°C, 167.45°C, and 189.43°C.

[0263] Table 5

[0264] Example 11 Preparation of the Crystalline Form F of Compound Represented by Formula (I)

[0265] 8 mg of the compound represented by formula (I) was added to 0.8 mL of methyl tert-butyl ether, stirred at room temperature for 2 days, centrifuged, and the solid dried in vacuo to obtain the product. X-ray powder diffraction analysis revealed an XRPD spectrum as shown in Figure 6 and characteristic peak positions as shown in Table 6. This product was defined as Form F of the compound represented by formula (I). The DSC spectrum showed endothermic peaks at 198.57°C and 208.23°C.

[0266] Table 6

[0267] Example 12 Preparation of the Crystalline Form of Compound G Represented by Formula (I)

[0268] 8 mg of the compound represented by formula (I) was added to 0.8 mL of dichloromethane, stirred at room temperature for 2 days, centrifuged, and the solid dried in vacuo to obtain the product. X-ray powder diffraction analysis revealed an XRPD spectrum as shown in Figure 7 and characteristic peak positions as shown in Table 7. This product was defined as Form G of the compound represented by formula (I). The DSC spectrum showed an endothermic peak at 177.46°C.

[0269] Table 7

[0270] Example 13 Preparation of Crystalline Form H of Compound Represented by Formula (I)

[0271] Crystal Form E of the compound represented by Formula (I) (Example 10) was heated to 180°C to obtain a product. X-ray powder diffraction analysis revealed an XRPD pattern as shown in Figure 8 , with characteristic peak positions as shown in Table 8. This product was defined as Crystal Form H of the compound represented by Formula (I). The DSC spectrum showed an endothermic peak at 193.39°C.

[0272] Table 8

[0273] Example 13 Preparation of the Crystalline Form of Compound I Represented by Formula (I)

[0274] 30 mg of the compound represented by formula (I) was dissolved in 0.3 mL of ethanol, stirred at room temperature to precipitate, centrifuged, and the solid dried in vacuo to obtain the product. X-ray powder diffraction analysis revealed an XRPD spectrum as shown in Figure 9 and characteristic peak positions as shown in Table 9. This product was defined as Form I of the compound represented by formula (I). The DSC spectrum showed an endothermic peak at 251.58°C.

[0275] Table 9

[0276] Example 14 Preparation of the Crystalline Form of Compound I Represented by Formula (I)

[0277] 8 mg of the compound represented by formula (I) was added to 0.8 mL of acetonitrile, stirred at room temperature for 2 days, filtered, and the solid was dried under vacuum to obtain a product. X-ray powder diffraction analysis showed that the product was Form I of the compound represented by formula (I).

[0278] Example 15 Preparation of the Crystalline Form of Compound I Represented by Formula (I)

[0279] 8 mg of the compound represented by formula (I) was dissolved in 0.04 mL of ethyl acetate, stirred at room temperature overnight, filtered, and the solid was dried under vacuum to obtain the product. X-ray powder diffraction analysis showed that the product was Form I of the compound represented by formula (I).

[0280] Example 16 Preparation of Crystal Form J of Compound Represented by Formula (I)

[0281] 60 mg of the compound represented by formula (I) was added to 0.5 mL of dichloromethane, stirred at room temperature overnight, filtered, and the solid dried under vacuum to obtain the product. X-ray powder diffraction analysis revealed an XRPD spectrum as shown in Figure 10 and characteristic peak positions as shown in Table 10. This product was defined as Form J of the compound represented by formula (I). The DSC spectrum showed an endothermic peak at 208.11°C.

[0282] Table 10

[0283] Example 17 Preparation of Crystalline Form K of Compound Represented by Formula (I)

[0284] 8 mg of the compound represented by formula (I) was dissolved in 0.04 mL of tetrahydrofuran, stirred at room temperature to precipitate, centrifuged, and the solid dried in vacuo to obtain the product. X-ray powder diffraction analysis revealed an XRPD pattern as shown in Figure 11, with the positions of its characteristic peaks shown in Table 11. This product was defined as Form K of the compound represented by formula (I). The DSC spectrum showed an endothermic peak at 208.48°C.

[0285] Table 11

[0286] Example 18 Preparation of Crystalline Form L of Compound Represented by Formula (I)

[0287] 60 mg of the compound represented by formula (I) was dissolved in 1 mL of ethyl acetate, stirred at room temperature to precipitate, filtered, and the solid dried in vacuo to obtain the product. X-ray powder diffraction analysis revealed an XRPD spectrum as shown in Figure 12, and the positions of its characteristic peaks are shown in Table 12. This product was defined as Form L of the compound represented by formula (I). The DSC spectrum showed endothermic peaks at 57.12°C, 137.09°C, and 193.02°C.

[0288] Table 12

[0289] Example 19 Preparation of Crystalline Form M of Compound Represented by Formula (I)

[0290] 60 mg of the compound represented by formula (I) was dispersed in 5 mL of isopropyl acetate, stirred at room temperature for 5 days, filtered, and the solid dried under vacuum to obtain the product. X-ray powder diffraction analysis revealed an XRPD spectrum as shown in Figure 13 and characteristic peak positions as shown in Table 13, defining the product as Form M of the compound represented by formula (I). The DSC spectrum showed endothermic peaks at 45.80°C, 130.75°C, and 193.53°C.

[0291] Table 13

[0292] Example 20 Preparation of Crystal Form N of Compound Represented by Formula (I)

[0293] 60 mg of the compound represented by formula (I) was dissolved in 1 mL of acetone, stirred at room temperature to precipitate, filtered, and the solid dried in vacuo to obtain the product. X-ray powder diffraction analysis revealed an XRPD spectrum as shown in Figure 14 and characteristic peak positions as shown in Table 14, defining the product as Form N of the compound represented by formula (I). The DSC spectrum showed endothermic peaks at 76.42°C and 143.82°C.

[0294] Table 14

[0295] Example 21 Preparation of Crystal Form O of Compound Represented by Formula (I)

[0296] 8 mg of the compound represented by formula (I) was dissolved in 0.12 mL of isopropanol, stirred at room temperature to precipitate, centrifuged, and the solid dried in vacuo to obtain the product. X-ray powder diffraction analysis revealed an XRPD spectrum as shown in Figure 15 , with the positions of its characteristic peaks shown in Table 15 . This product was defined as the O-form of the compound represented by formula (I). The DSC spectrum showed endothermic peaks at 120.14°C and 177.32°C.

[0297] Table 15

[0298] Example 22 Crystal Stability Study

[0299] The A, B, C, I, J, and K crystal forms of the compound represented by formula (I) were exposed and laid flat, and the stability of the samples was investigated under light (4500 Lux), high temperature (40°C, 60°C), and high humidity (RH 75%, RH 92.5%) conditions. The sampling period was 30 days. The results are shown in Table 16.

[0300] Table 16

[0301] Conclusion: The influencing factor experiment shows that the physical and chemical properties of A, B, C, I, J, and K crystal forms are good when placed under high temperature and high humidity conditions for 30 days.

[0302] Example 23 Long-term / accelerated stability

[0303] The stability of the A, B, C, I, J, and K crystal forms of the compound represented by formula (I) were investigated at 25°C / 60% RH and 40°C / 75% RH, respectively. The results are shown in Table 17.

[0304] Table 17

[0305] Conclusion: Long-term accelerated experiments show that Forms A, B, C, I, J, and K have good physical and chemical stability when placed under 25°C / 60RH and 40°C / 75RH conditions for 3 or 6 months.

Claims

1. A crystalline form A of a compound represented by formula (I), having an X-ray powder diffraction spectrum with characteristic peaks at 2θ angles of 9.2, 13.1, 14.5, 16.6, 17.7, and 21.

5.

2. The crystal form A of the compound represented by formula (I) according to claim 1, has an X-ray powder diffraction spectrum with characteristic peaks at 2θ angles of 6.6, 9.2, 10.1, 10.7, 13.1, 14.5, 15.1, 16.6, 17.7, 19.6, 20.9, 21.5, 22.2, 24.0, 24.5, 24.9, 25.9, 27.2, 28.6, and 30.

2.

3. The crystal form A of the compound represented by formula (I) according to claim 1, whose X-ray powder diffraction pattern is shown in Figure 1.

4. A crystal form B of the compound represented by formula (I), whose X-ray powder diffraction spectrum has characteristic peaks at 2θ angles of 8.8, 12.7, 14.6, 14.9, 16.2, 18.0, and 21.

4.

5. The crystalline form B of the compound represented by formula (I) according to claim 4, has an X-ray powder diffraction spectrum with characteristic peaks at 2θ angles of 7.3, 8.8, 10.1, 10.6, 12.7, 14.6, 14.9, 16.2, 16.6, 17.3, 18.0, 19.2, 20.0, 21.4, 22.2, 24.4, 25.0, 25.7, 28.7, and 29.

4.

6. The crystal form B of the compound represented by formula (I) according to claim 4, whose X-ray powder diffraction pattern is shown in Figure 2.

7. A crystal form C of the compound represented by formula (I), whose X-ray powder diffraction spectrum has characteristic peaks at 2θ angles of 5.7, 7.6, 11.1, 11.4, 17.9, and 19.

3.

8. The crystalline form C of the compound represented by formula (I) according to claim 7, has an X-ray powder diffraction spectrum with characteristic peaks at 2θ angles of 5.7, 6.4, 6.9, 7.6, 11.1, 11.4, 12.7, 13.9, 15.9, 17.2, 17.9, 19.3, 23.9, 24.4, 25.7, and 26.

4.

9. The crystal form C of the compound represented by formula (I) according to claim 7, whose X-ray powder diffraction pattern is shown in Figure 3.

10. A crystalline form I of a compound represented by formula (I), having an X-ray powder diffraction spectrum with characteristic peaks at 2θ angles of 6.9, 15.3, 16.6, and 20.

1.

11. The crystalline form I of the compound represented by formula (I) according to claim 10, having an X-ray powder diffraction spectrum with characteristic peaks at 2θ angles of 5.6, 6.9, 8.3, 9.6, 11.4, 13.9, 15.3, 16.6, 18.4, 20.1, 20.6, 21.1, 22.1, 23.1, 24.5, 26.0, and 33.

0.

12. The crystal form I of the compound represented by formula (I) according to claim 10, whose X-ray powder diffraction pattern is shown in Figure 9.

13. A J crystal form of the compound represented by formula (I), having an X-ray powder diffraction spectrum with characteristic peaks at 2θ angles of 7.2, 7.7, 8.5, 9.8, 10.6, and 14.

9.

14. The crystalline form J of the compound represented by formula (I) according to claim 13, having an X-ray powder diffraction spectrum with characteristic peaks at 2θ angles of 7.2, 7.7, 8.5, 9.8, 10.6, 12.2, 13.5, 14.9, 15.7, 17.1, 17.9, 19.1, 20.0, 20.5, 21.6, 23.3, 24.3, 25.1, 25.7, and 28.

3.

15. The J crystal form of the compound represented by formula (I) according to claim 13, whose X-ray powder diffraction pattern is shown in Figure 10.

16. A K-type crystal of the compound represented by formula (I), having an X-ray powder diffraction spectrum with characteristic peaks at 2θ angles of 5.3, 6.7, 10.7, 12.3, 13.5, 14.8, 18.1, 20.6, 21.3, and 27.

3.

17. The K crystal form of the compound represented by formula (I) according to claim 16, whose X-ray powder diffraction pattern is shown in Figure 11.

18. The crystalline form of the compound of formula (I) according to any one of claims 1 to 17, wherein the error range of the 2θ angle is ±0.

2.

19. A method for preparing Form A of the compound of formula (I) according to any one of claims 1 to 3 and 18, comprising: Mixing the compound represented by formula (I) with an appropriate amount of solvent, cooling and crystallizing; The solvent is selected from one or more of methanol and water / methanol.

20. A method for preparing the crystal form B of the compound represented by formula (I) as claimed in any one of claims 4 to 6 and 18, comprising: Mixing the compound represented by formula (I) with an appropriate amount of solvent, cooling and crystallizing; The solvent is selected from one or more of ethanol, isopropanol, n-propanol, ethyl acetate / ethanol, and ethyl acetate / n-heptane.

21. A method for preparing the crystal form C of the compound represented by formula (I) as claimed in any one of claims 7 to 9 and 18, comprising: The compound represented by formula (I) is mixed with an appropriate amount of solvent and crystallized; the solvent is acetonitrile.

22. A method for preparing the crystalline form I of the compound represented by formula (I) as claimed in any one of claims 10 to 12 and 18, comprising: The compound represented by formula (I) is mixed with an appropriate amount of solvent and stirred at room temperature for crystallization; the solvent is selected from one or more of methanol, ethanol, n-propanol, water / methanol, water / ethanol, water / isopropanol, acetonitrile / methanol, acetone, water / acetone, 2-butanone, acetonitrile, ethyl acetate, isopropyl acetate, n-heptane, tetrahydrofuran / ethanol, ethyl acetate / ethanol, ethyl acetate / n-heptane, isopropyl ether, and methyl tert-butyl ether.

23. A method for preparing the J crystal form of the compound represented by formula (I) as claimed in any one of claims 13 to 15 and 18, comprising: Mixing the compound represented by formula (I) with an appropriate amount of solvent and crystallizing; The solvent is dichloromethane.

24. A method for preparing the K crystal form of the compound represented by formula (I) according to any one of claims 16 to 18, comprising: The compound represented by formula (I) is mixed with an appropriate amount of solvent and crystallized; the solvent is tetrahydrofuran.

25. A pharmaceutical composition comprising the following components: (a) Form A of the compound of formula (I) according to any one of claims 1 to 3 and 18, Form B of the compound of formula (I) according to any one of claims 4 to 6 and 18, Form C of the compound of formula (I) according to any one of claims 7 to 9 and 18, Form I of the compound of formula (I) according to any one of claims 10 to 12 and 18, Form J of the compound of formula (I) according to any one of claims 13 to 15 and 18, or Form K of the compound of formula (I) according to any one of claims 16 to 18; and (b) optionally selected from pharmaceutically acceptable carriers, diluents or excipients.

26. A method for preparing a pharmaceutical composition, comprising: (a) Form A of the compound of formula (I) according to any one of claims 1 to 3 and 18, Form B of the compound of formula (I) according to any one of claims 4 to 6 and 18, Form C of the compound of formula (I) according to any one of claims 7 to 9 and 18, Form I of the compound of formula (I) according to any one of claims 10 to 12 and 18, Form J of the compound of formula (I) according to any one of claims 13 to 15 and 18, or Form K of the compound of formula (I) according to any one of claims 16 to 18; and (b) optionally mixing with a pharmaceutically acceptable carrier, diluent or excipient.

27. Use of the crystal form A of the compound represented by formula (I) according to any one of claims 1-3 and 18, the crystal form B of the compound represented by formula (I) according to any one of claims 4-6 and 18, the crystal form C of the compound represented by formula (I) according to any one of claims 7-9 and 18, the crystal form I of the compound represented by formula (I) according to any one of claims 10-12 and 18, the crystal form J of the compound represented by formula (I) according to any one of claims 13-15 and 18, the crystal form K of the compound represented by formula (I) according to any one of claims 16-18, or the composition according to claim 25 in the preparation of a medicament for inhibiting KRAS G12D.

28. Use of Form A of the compound represented by formula (I) according to any one of claims 1-3 and 18, Form B of the compound represented by formula (I) according to any one of claims 4-6 and 18, Form C of the compound represented by formula (I) according to any one of claims 7-9 and 18, Form I of the compound represented by formula (I) according to any one of claims 10-12 and 18, Form J of the compound represented by formula (I) according to any one of claims 13-15 and 18, Form K of the compound represented by formula (I) according to any one of claims 16-18, or the composition according to claim 25 in the preparation of a medicament for treating and / or preventing a disease or condition, wherein the disease or condition is cancer.

29. The method of claim 28, wherein the disease or condition is selected from the group consisting of brain cancer, thyroid cancer, head and neck cancer, nasopharyngeal cancer, pharyngeal cancer, oral cancer, salivary gland cancer, esophageal cancer, gastric cancer, lung cancer, liver cancer, kidney cancer, pancreatic cancer, gallbladder cancer, bile duct cancer, colorectal cancer, small intestine cancer, gastrointestinal stromal tumors, urothelial cancer, urethral cancer, bladder cancer, breast cancer, vaginal cancer, ovarian cancer, endometrial cancer, cervical cancer, fallopian tube cancer, testicular cancer, prostate cancer, hemangioma, leukemia, lymphoma, myeloma, skin cancer, lipoma, bone cancer, soft tissue sarcoma, neurofibroma, glioma, neuroblastoma and glioblastoma; preferably selected from the group consisting of pancreatic cancer, colorectal cancer and non-small cell lung cancer.

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