Salt crystal form of aromatic nitrogen-containing compound and preparation method therefor

Abstract

The present disclosure relates to a salt crystal form of an aromatic nitrogen-containing compound and a preparation method therefor. Specifically, the present disclosure relates to a salt crystal form of a compound represented by general formula (I), a preparation method therefor, a pharmaceutical composition containing a therapeutically effective amount of the salt crystal form, and a pharmaceutical use thereof for treating a disease or condition associated with a specific target.

Description

A salt crystal form of an aromatic nitrogen-containing compound and its preparation method Technical Field

[0001] This disclosure pertains to the pharmaceutical field and relates to a salt crystal form of an aromatic nitrogen-containing compound, a method for its preparation, a pharmaceutical composition, and its pharmaceutical uses. Background Technology

[0002] Cells in any organism possess multiple DNA repair mechanisms to maintain genome stability and integrity, which is crucial for survival. Mismatch repair (MMR) is a highly conserved pathway that plays a key role in maintaining genome stability by correcting errors that occur during DNA replication, recombination, and repair. (Pecina-Slaus, N., Kafka, A., Salamon, I. & Bukovac, A. Mismatch Repair Pathway, Genome Stability and Cancer. Front Mol. Biosci. 7, 122 (2020)). Defects in the MMR mechanism can lead to genome hypermutation and instability, manifested as a high frequency of insertions and deletions of short repetitive DNA sequences (microsatellites) throughout the genome; this phenomenon is called microsatellite instability (MSI). (Kim, TM, Laird, PW & Park, PJ. The landscape of microsatellite instability in colorectal and endometrial cancer genomes. Cell 155, 858–868 (2013); Vernole, P. et al. Common fragile sites in colon cancer cell lines: role of mismatch repair, RAD51 and poly(ADP-ribose)polymerase-1. Mutat. Res 712, 40–48 (2011)). Microsatellites are generated by DNA polymerase slippage during replication or mismatch repair and are prone to misplacement and scaffold shift mutations, which can be identified and corrected by MMR processes in normal cells. However, in cancer cells lacking normal mismatch repair mechanisms, errors occurring during replication or repair accumulate, increasing the rate of genomic mutations (Lower, SS, McGurk, MP, Clark, AG, et al. Satellite DNA evolution: old ideas, new approaches. Curr. Opin. Genet. Dev., 2018; 49:70-78).High-frequency microsatellite deletions and insertions can lead to microsatellite instability (MSI), which in turn contributes to 10-30% of ovarian, colon, gastric, and endometrial cancers (Aaltonen, LA et al. Clues to the pathogenesis of familial colorectal cancer, Science 260, 812-816 (1993), Bonneville R et al., Landscape of Microsatellite Instability Across 39 Cancer Types. JCO Precis Oncol. 1: PO. 17. 00073 (2017)).Compared to MSS cancers, patients with high MSI (MSI-H) cancers exhibit better overall prognosis and reduced metastatic potential, higher tumor mutational burden, and greater immunogenicity (Kang, S. et al. The significance of microsatellite instability in colorectal cancer after controlling for clinicopathological factors. Med. (Baltim.) 97, e0019 (2018)). MSI-H tumors often develop resistance to immunotherapy and chemotherapy (Le, D.T. et al. Phase II Open-Label Study of Pembrolizumab in Treatment-Refractory, Microsatellite Instability-High / Mismatch Repair-Deficient Metastatic Colorectal Cancer: KEYNOTE-164. J. Clin. Oncol. 38, 11–19 (2020), Overman, MJ et al. Nivolumab in patients with metastatic DNA mismatch repair-deficient or microsatellite instability-high colorectal cancer (CheckMate 142): an open-label, multicentre, phase 2 study. Lancet Oncol. 18, 1182–1191 (2017), Fuca, G. et al. Ascites and resistance to immune checkpoint inhibition in dMMR / MSI-H metastatic colorectal and gastric cancers. J. Immunother. Cancer 10, 4001 (2022)).

[0003] Recently, multiple independent large-scale functional genomic screenings using more than 300 human cancer cell lines have revealed that Werner syndrome RecQ helicase (WRN) is selectively essential for the survival of MSI-H cells (Behan, FM et al. Prioritization of cancer therapeutic targets using CRISPR-Cas9 screens. Nature 568, 511–516 (2019), McDonald ER et al., Project DRIVE: A Compendium of Cancer Dependencies and Synthetic Lethal Relationships Uncovered by Large-Scale, Deep RNAi Screening. Cell 170(3):577-592 (2017), Chan, EM et al. WRN helicase is a synthetic lethal target in microsatellite unstable cancers. Nature 568, 551–556 (2019)). WRN is one of the five RecQ-like helicases in humans. It is a multifunctional enzyme with helicase and exonuclease activities and plays an important role in multiple pathways.

[0004] DNA repair and maintenance of genome integrity include DNA replication, transcription, DNA repair and telomere maintenance (Bohr, VARising from the RecQ-age: the role of human RecQ helicases in genome maintenance. Trends Biochem Sci. 33, 609–620 (2008); Singh, DK, Ahn, B. & Bohr, VARoles of RECQ helicases in recombination based DNA repair, genomic stability and aging. Biogerontology 10, 235–252 (2009); Rossi, ML, Ghosh, AK & Bohr, VARoles of Werner syndrome protein in protection of genome integrity. DNA Repair (Amst.) 9, 331–344 (2010)).

[0005] The absence of WRN leads to various defects in cellular and genomic structures, including cell cycle arrest, DNA breaks, mitotic defects, chromosome fragmentation, and MSI (but not MSS) cell apoptosis (Behan, FM et al. Prioritization of cancer therapeutic targets using CRISPR-Cas9 screens. Nature 568, 511–516 (2019), McDonald ER et al., Project DRIVE: A Compendium of Cancer Dependencies and Synthetic Lethal Relationships Uncovered by Large-Scale, Deep RNAi Screening. Cell 170(3):577-592 (2017), Chan, EM et al. WRN helicase is a synthetic lethal target in microsatellite unstable cancers. Nature 568, 551–556 (2019)). Furthermore, studies on loss-of-function mutations and deletions of WRN have further clarified that the helicase function of WRN is indispensable for the survival of MSI cells (Lieb, S. et al. Werner syndrome helicase is a selective vulnerability of microsatellite instability-high tumor cells. Elife 8, e43333 (2019)). These results clearly indicate that targeting WRN, especially the helicase domain, may be a promising strategy for treating high MSI cancers.

[0006] Patent PCT / CN2024 / 102582 protects a class of aromatic nitrogen-containing compounds. Given the importance of studying the salt form and crystal form of drugs for clinical research, and in order to improve the solubility and solid stability of the product, reduce storage costs, extend the product cycle, and enhance the bioavailability of the product, this disclosure studies the salt crystal form of the aforementioned compounds. Summary of the Invention

[0007] All contents relating to patent PCT / CN2024 / 102582 are incorporated herein by reference.

[0008] The purpose of this disclosure is to provide an acid salt crystal form or a basic salt crystal form of the compound shown in formula (I), wherein the structure of the compound is as follows:

[0009] in,

[0010] X1 and X2 are independently selected from CH or N;

[0011] R1 is selected from H, halogen, hydroxyl, C1-C6 alkyl, C1-C6 alkoxy, C1-C6 hydroxyalkyl, and C1-C6 haloalkyl;

[0012] R2 is selected from H, halogen, hydroxyl, C1-C6 alkyl, C1-C6 alkoxy, C1-C6 hydroxyalkyl, and C1-C6 haloalkyl;

[0013] R3 is selected from H, halogen, hydroxyl, C1-C6 alkyl, C1-C6 alkoxy, C1-C6 hydroxyalkyl, and C1-C6 haloalkyl;

[0014] R4 is selected from H, halogen, hydroxyl, C1-C6 alkyl, C1-C6 alkoxy, C1-C6 hydroxyalkyl, and C1-C6 haloalkyl;

[0015] R5 is selected from H, halogen, hydroxyl, C1-C6 alkyl, C1-C6 alkoxy, C1-C6 hydroxyalkyl, and C1-C6 haloalkyl;

[0016] R6 is selected from H, halogen, hydroxyl, C1-C6 alkyl, C1-C6 alkoxy, C1-C6 hydroxyalkyl, and C1-C6 haloalkyl;

[0017] R7 is selected from H, halogen, hydroxyl, C1-C6 alkyl, C1-C6 alkoxy, C1-C6 hydroxyalkyl, and C1-C6 haloalkyl;

[0018] R8 is selected from H, halogen, hydroxyl, C1-C6 alkyl, C1-C6 alkoxy, C1-C6 hydroxyalkyl, and C1-C6 haloalkyl;

[0019] R9 is selected from H, halogen, hydroxyl, C1-C6 alkyl, C1-C6 alkoxy, C1-C6 hydroxyalkyl, and C1-C6 haloalkyl;

[0020] R 10 Selected from heterocyclic groups, wherein the heterocyclic group is a 5- or 6-membered fully saturated or partially unsaturated group, comprising a cyclic carbon atom and one or two cyclic heteroatoms selected from N, O and S respectively;

[0021] The acid is an inorganic or organic acid, wherein the inorganic acid is selected from hydrochloric acid, sulfuric acid, nitric acid, hydrobromic acid, hydrofluoric acid, hydroiodic acid, or phosphoric acid; the organic acid is selected from 2,5-dihydroxybenzoic acid, 1-hydroxy-2-naphtholic acid, acetic acid, dichloroacetic acid, trichloroacetic acid, acetoxyxamic acid, adipic acid, benzenesulfonic acid, 4-chlorobenzenesulfonic acid, benzoic acid, 4-acetaminobenzoic acid, 4-aminobenzoic acid, decanoic acid, hexanoic acid, caprylic acid, cinnamic acid, citric acid, cyclohexanesulfonic acid, camphorsulfonic acid, aspartic acid, camphoric acid, gluconic acid, glucuronic acid, glutamic acid, isoascorbic acid, lactic acid, malic acid, mandelic acid, pyroglutamic acid, and alcohol. Sarcoic acid, dodecyl sulfuric acid, dibenzoyl tartaric acid, ethane-1,2-disulfonic acid, ethanesulfonic acid, formic acid, fumaric acid, galactobionic acid, gentian acid, glutaric acid, 2-ketoglutaric acid, glycolic acid, hippuric acid, hydroxyethyl sulfonic acid, lactobionic acid, ascorbic acid, aspartic acid, lauric acid, camphoric acid, maleic acid, malonic acid, methanesulfonic acid, 1,5-naphthalenedisulfonic acid, naphthalene-2-sulfonic acid, nicotinic acid, oleic acid, orotic acid, oxalic acid, palmitic acid, dihydroxynaphthalic acid, propionic acid, salicylic acid, 4-aminosalicylic acid, sebacic acid, stearic acid, succinic acid, thiocyanate, undecanoic acid, trifluoroacetic acid, benzenesulfonic acid, p-methylbenzenesulfonic acid, or L-malic acid;

[0022] The base is an organic or inorganic base. The organic base is selected from sodium methoxide, potassium ethoxide, trimethylamine, diethylamine, triethylamine, triethanolamine, pyridine, piperidine, morpholine, diisopropylethylamine, diisopropylaminolithium, diethylaminolithium, bis(trimethylsilyl)aminolithium, potassium acetate, sodium acetate, isopropylcyclohexylaminolithium or mixtures thereof. The inorganic base is selected from potassium phosphate, potassium phosphate trihydrate, potassium phosphate dihydrate, potassium phosphate monohydrate, sodium bicarbonate, potassium bicarbonate, sodium carbonate, cesium carbonate, potassium hydroxide, sodium hydroxide, potassium hydride, sodium hydride, lithium hydroxide or mixtures thereof.

[0023] In a further preferred embodiment of this disclosure,

[0024] X1 and X2 are N;

[0025] R1 is selected from C1-C3 alkyl groups;

[0026] R2 is selected from F, Cl, Br, and I;

[0027] R3 is selected from C 1-3 Halogenated alkyl groups, preferably CF3;

[0028] R4 is selected from hydrogen;

[0029] R5 is selected from OH;

[0030] R6 is selected from C1-C3 alkyl groups;

[0031] R7 is selected from hydrogen;

[0032] R8 is selected from H or C1-C3 alkyl groups;

[0033] R9 is selected from C1-C3 alkyl groups;

[0034] R10 is selected from

[0035] In a further preferred embodiment of this disclosure, the compound is selected from either compound 1 or compound 2:

[0036] In a further preferred embodiment of this disclosure, the salt crystal form is a solvent-containing or solvent-free crystal form, wherein the solvent is selected from water, methanol, acetone, ethyl acetate, acetonitrile, ethanol, 88% acetone, 2-methyltetrahydrofuran, dichloromethane, 1,4-dioxane, benzene, toluene, isopropanol, n-butanol, isobutanol, N,N-dimethylformamide, N,N-dimethylacetamide, dimethyl sulfoxide, n-propanol, tert-butanol, 2-butanone, 3-pentanone, n-heptane, ethyl formate, isopropyl acetate, cyclohexane, methyl tert-butyl ether, or isopropyl ether; preferably, the number of solvents is 0.2-3, more preferably 0.2, 0.5, 1, 1.5, 2, 2.5, or 3, and most preferably 0.5, 1, 2, or 3; the crystal form is more preferably anhydrous.

[0037] In a further preferred embodiment of this disclosure, the acid salt crystal form is a sulfate crystal form, and the basic salt crystal form is a potassium salt crystal form or a sodium salt crystal form.

[0038] In a further preferred embodiment of this disclosure, the crystal form is sulfate crystal form A of compound 1, wherein:

[0039] The powder X-ray diffraction pattern of sulfate crystal form A of compound 1 has a characteristic peak at 2θ of 13.8 ± 0.2°, or at 2θ of 16.6 ± 0.2°, or at 2θ of 17.7 ± 0.2°, or at 2θ of 15.3 ± 0.2°, or at 2θ of 9.0 ± 0.2°, or at 2θ of 25.1 ± 0.2°, or at 2θ of 22.6 ± 0.2°, or at 2θ of 21. The characteristic peak is present at 0 ± 0.2°, or at 2θ = 20.2 ± 0.2°, or at 2θ = 23.5 ± 0.2°, or at 2θ = 9.9 ± 0.2°, or at 2θ = 11.1 ± 0.2°, or at 2θ = 27.8 ± 0.2°, or at 2θ = 14.6 ± 0.2°; preferably, the characteristic peak is present at any 2, 4, 6, 8, 10, or 12 of these locations.

[0040] In a further preferred embodiment of this disclosure, the powder X-ray diffraction pattern of sulfate crystal form A of compound 1 has one or more characteristic peaks at 13.8±0.2°, 16.6±0.2°, 17.7±0.2°, or 15.3±0.2°; preferably including 2-4 such peaks, more preferably including 3-4 such peaks, and most preferably including 4 such peaks; optionally, further, it may also include one or more characteristic peaks at 2θ of 9.0±0.2°, 25.1±0.2°, 22.6±0.2°, 21.0±0.2°, or 20.2±0.2°, preferably including 2, 3, 4, or 5 such peaks; for example, the X-ray powder diffraction pattern of sulfate crystal form A of compound 1 has diffraction peaks at the following positions at 2θ:

[0041] At 13.8±0.2°, 16.6±0.2° and 17.7±0.2°,

[0042] Alternatively, at 13.8±0.2°, 16.6±0.2°, 17.7±0.2°, and 15.3±0.2°,

[0043] Alternatively, at 13.8±0.2°, 16.6±0.2°, 17.7±0.2°, 15.3±0.2°, and 9.0±0.2°;

[0044] In a further preferred embodiment of this disclosure, the X-ray powder diffraction pattern of sulfate crystal form A of compound 1 has diffraction peaks at one or more of the following 2θ values: 13.8±0.2°, 16.6±0.2°, 17.7±0.2°, 15.3±0.2°, 9.0±0.2°, 25.1±0.2°, 22.6±0.2°, 21.0±0.2°, 20.2±0.2°, 23.5±0.2°, 9.9±0.2°, 11.1±0.2°, 27.8±0.2°, and 14.6±0.2°; preferably, it includes diffraction peaks at any of 4, 6, 8, or 10 of these values; for example, the X-ray powder diffraction pattern of sulfate crystal form A of compound 1 has diffraction peaks at the following 2θ positions:

[0045] At 13.8±0.2°, 16.6±0.2°, 17.7±0.2°, 15.3±0.2°, 9.0±0.2°, 25.1±0.2°, 22.6±0.2°, and 21.0±0.2°,

[0046] Alternatively, at 13.8±0.2°, 16.6±0.2°, 17.7±0.2°, 15.3±0.2°, 9.0±0.2°, 25.1±0.2°, 22.6±0.2°, 21.0±0.2°, 20.2±0.2°, and 23.5±0.2°.

[0047] The characteristic X-ray diffraction peaks of sulfate crystal form A of compound 1 are shown in Table 9.

[0048] In a further preferred embodiment of this disclosure, the X-ray powder diffraction pattern of sulfate crystal form A of compound 1 is shown in Figure 1.

[0049] In a further preferred embodiment of this disclosure, the 2θ error between the positions of the top ten diffraction peaks with the highest relative intensities in the X-ray powder diffraction pattern of the sulfate crystal form A of compound 1 and the corresponding diffraction peaks in Figure 1 is ±0.2° to ±0.5°, preferably ±0.2° to ±0.3°, and more preferably ±0.2°.

[0050] In a further preferred embodiment of this disclosure, the crystal form is potassium salt crystal form A of compound 1, wherein:

[0051] The powder X-ray diffraction pattern of potassium salt crystal form A of compound 1 has a characteristic peak at 2θ of 5.7 ± 0.2°, or at 2θ of 16.0 ± 0.2°, or at 2θ of 21.3 ± 0.2°, or at 2θ of 17.0 ± 0.2°, or at 2θ of 13.9 ± 0.2°, or at 2θ of 25.5 ± 0.2°, or at 2θ of 29.7 ± 0.2°, or at 2θ of 28.6 ± 0.2°. The peak has a characteristic peak at 2θ = 34.0 ± 0.2°, or at 2θ = 20.5 ± 0.2°, or at 2θ = 29.2 ± 0.2°, or at 2θ = 37.3 ± 0.2°, or at 2θ = 26.2 ± 0.2°, or at 2θ = 30.9 ± 0.2°, or at 2θ = 11.3 ± 0.2°; preferably, it includes any 2, 4, 6, 8, 10 or 12 of these peaks.

[0052] In a further preferred embodiment of this disclosure, the powder X-ray diffraction pattern of potassium salt crystal form A of compound 1 has one or more characteristic peaks at 5.7±0.2°, 16.0±0.2°, 21.3±0.2°, or 17.0±0.2°; preferably including 2-4 such peaks, more preferably including 3-4 such peaks, and most preferably including 4 such peaks; optionally, further, it may also include one or more characteristic peaks at 2θ of 13.9±0.2°, 25.5±0.2°, 29.7±0.2°, 28.6±0.2°, or 34.0±0.2°, preferably including 2, 3, 4, or 5 such peaks; for example, the X-ray powder diffraction pattern of potassium salt crystal form A of compound 1 has diffraction peaks at the following positions at 2θ:

[0053] At 5.7±0.2°, 16.0±0.2°, and 21.3±0.2°,

[0054] Alternatively, at 5.7±0.2°, 16.0±0.2°, 21.3±0.2°, and 17.0±0.2°,

[0055] Alternatively, at 5.7±0.2°, 16.0±0.2°, 21.3±0.2°, 17.0±0.2°, 13.9±0.2°, and 25.5±0.2°.

[0056] In a further preferred embodiment of this disclosure, the powder X-ray diffraction pattern of potassium salt crystal form A of compound 1 has characteristic peaks at one or more of the following 2θ values: 5.7±0.2°, 16.0±0.2°, 21.3±0.2°, 17.0±0.2°, 13.9±0.2°, 25.5±0.2°, 29.7±0.2°, 28.6±0.2°, 34.0±0.2°, 20.5±0.2°, 29.2±0.2°, 37.3±0.2°, 26.2±0.2°, 30.9±0.2°, and 11.3±0.2°. Preferably, the diffraction peaks include any 4, 6, 8, or 10 of these values. For example, the X-ray powder diffraction pattern of potassium salt crystal form A has diffraction peaks at the following 2θ positions:

[0057] At 5.7±0.2°, 16.0±0.2°, 21.3±0.2°, 17.0±0.2°, 13.9±0.2°, 25.5±0.2°, 29.7±0.2°, and 28.6±0.2°,

[0058] Alternatively, at 5.7±0.2°, 16.0±0.2°, 21.3±0.2°, 17.0±0.2°, 13.9±0.2°, 25.5±0.2°, 29.7±0.2°, 28.6±0.2°, 34.0±0.2°, and 20.5±0.2°.

[0059] The characteristic X-ray diffraction peaks of potassium salt crystal form A of compound 1 are shown in Table 10.

[0060] In a further preferred embodiment of this disclosure, the X-ray powder diffraction pattern of potassium salt crystal form A of compound 1 is shown in Figure 2.

[0061] In a further preferred embodiment of this disclosure, the 2θ error between the positions of the top ten diffraction peaks with the highest relative intensities in the X-ray powder diffraction pattern of potassium salt crystal form A of compound 1 and the corresponding diffraction peaks in Figure 2 is ±0.2° to ±0.5°, preferably ±0.2° to ±0.3°, and more preferably ±0.2°.

[0062] In a further preferred embodiment of this disclosure, the crystal form is potassium salt crystal form B of compound 1, wherein:

[0063] The powder X-ray diffraction pattern of potassium salt crystal form B of compound 1 has a characteristic peak at 2θ of 5.2 ± 0.2°, or at 2θ of 7.4 ± 0.2°, or at 2θ of 14.5 ± 0.2°, or at 2θ of 16.1 ± 0.2°, or at 2θ of 18.7 ± 0.2°, or at 2θ of 21.8 ± 0.2°, or at 2θ of 11.7 ± 0.2°, or at 2θ of 10.0 ± 0.2°, or at 2θ of 26.0 ± 0.2°, or at 2θ of 20.4 ± 0.2°; preferably, it includes any 2, 4, 6, 8, or 10 characteristic peaks.

[0064] In a further preferred embodiment of this disclosure, the powder X-ray diffraction pattern of potassium salt form B of compound 1 has one or more characteristic peaks at 5.2±0.2°, 7.4±0.2°, 14.5±0.2°, or 16.1±0.2°; preferably including 2-4 such peaks, more preferably including 3-4 such peaks, and most preferably including 4 such peaks; optionally, further, it may also include one or more characteristic peaks at 2θ of 18.7±0.2°, 21.8±0.2°, 11.7±0.2°, 10.0±0.2°, or 26.0±0.2°, preferably including 2, 3, 4, or 5 such peaks; for example, the X-ray powder diffraction pattern of potassium salt form B of compound 1 has diffraction peaks at the following positions at 2θ:

[0065] At 5.2±0.2°, 7.4±0.2°, and 14.5±0.2°,

[0066] Alternatively, at 5.2±0.2°, 7.4±0.2°, 14.5±0.2°, and 16.1±0.2°,

[0067] Alternatively, at 5.2±0.2°, 7.4±0.2°, 14.5±0.2°, 16.1±0.2°, 18.7±0.2°, and 21.8±0.2°.

[0068] In a further preferred embodiment of this disclosure, the powder X-ray diffraction pattern of potassium salt crystal form B of compound 1 has characteristic peaks at one or more of the following 2θ values: 5.2±0.2°, 7.4±0.2°, 14.5±0.2°, 16.1±0.2°, 18.7±0.2°, 21.8±0.2°, 11.7±0.2°, 10.0±0.2°, 26.0±0.2°, and 20.4±0.2°. Preferably, the diffraction peaks include any 4, 6, 8, or 10 of these values. For example, the X-ray powder diffraction pattern of potassium salt crystal form B of compound 1 has diffraction peaks at the following 2θ positions:

[0069] At angles of 5.2±0.2°, 7.4±0.2°, 14.5±0.2°, 16.1±0.2°, 18.7±0.2°, 21.8±0.2°, 11.7±0.2°, and 10.0±0.2°,

[0070] Alternatively, at 5.2±0.2°, 7.4±0.2°, 14.5±0.2°, 16.1±0.2°, 18.7±0.2°, 21.8±0.2°, 11.7±0.2°, 10.0±0.2°, 26.0±0.2°, and 20.4±0.2°.

[0071] The characteristic X-ray diffraction peaks of potassium salt crystal form B of compound 1 are shown in Table 11.

[0072] In a further preferred embodiment of this disclosure, the X-ray powder diffraction pattern of potassium salt crystal form B of compound 1 is shown in Figure 3.

[0073] In a further preferred embodiment of this disclosure, the 2θ error between the positions of the top ten diffraction peaks with the highest relative intensities in the X-ray powder diffraction pattern of potassium salt crystal form B of compound 1 and the corresponding diffraction peaks in Figure 3 is ±0.2° to ±0.5°, preferably ±0.2° to ±0.3°, and more preferably ±0.2°.

[0074] In a further preferred embodiment of this disclosure, the crystal form is sodium salt crystal form 1 of compound 1, wherein:

[0075] The powder X-ray diffraction pattern of sodium salt crystal form 1 of compound 1 has a characteristic peak at 2θ = 8.9 ± 0.2°, or at 2θ = 10.0 ± 0.2°, or at 2θ = 20.1 ± 0.2°, or at 2θ = 9.6 ± 0.2°, or at 2θ = 16.8 ± 0.2°, or at 2θ = 15.3 ± 0.2°, or at 2θ = 31.0 ± 0.2°, or at 2θ = 26.7 ± 0.2°, or at 2θ = 11.4 ± 0.2°, or at 2θ = 24.2 ± 0.2°; preferably, it includes any 2, 4, 6, 8 or 10 characteristic peaks.

[0076] In a further preferred embodiment of this disclosure, the powder X-ray diffraction pattern of sodium salt crystal form 1 of compound 1 has one or more characteristic peaks at 8.9±0.2°, 10.0±0.2°, 20.1±0.2°, and 9.6±0.2°; preferably including 2-4 such peaks, more preferably including 3-4 such peaks, and most preferably including 4 such peaks; optionally, further, it may also include one or more characteristic peaks at 2θ of 16.8±0.2°, 15.3±0.2°, 31.0±0.2°, 26.7±0.2°, or 11.4±0.2°, preferably including 2, 3, 4, or 5 such peaks; for example, the X-ray powder diffraction pattern of sodium salt crystal form 1 of compound 1 has diffraction peaks at the following positions at 2θ:

[0077] At 8.9±0.2°, 10.0±0.2°, and 20.1±0.2°,

[0078] Alternatively, at 8.9±0.2°, 10.0±0.2°, 20.1±0.2°, and 9.6±0.2°,

[0079] Alternatively, at 8.9±0.2°, 10.0±0.2°, 20.1±0.2°, 9.6±0.2°, 16.8±0.2°, and 15.3±0.2°.

[0080] In a further preferred embodiment of this disclosure,

[0081] The powder X-ray diffraction pattern of sodium salt crystal form 1 of compound 1 has characteristic peaks at one or more of the following positions at 2θ: 8.9±0.2°, 10.0±0.2°, 20.1±0.2°, 9.6±0.2°, 16.8±0.2°, 15.3±0.2°, 31.0±0.2°, 26.7±0.2°, 11.4±0.2°, and 24.2±0.2°; preferably, diffraction peaks are found at any of 4, 6, 8, or 10 of these positions. For example, the X-ray powder diffraction pattern of sodium salt crystal form 1 of compound 1 has diffraction peaks at the following positions at 2θ:

[0082] At angles of 8.9±0.2°, 10.0±0.2°, 20.1±0.2°, 9.6±0.2°, 16.8±0.2°, 15.3±0.2°, 31.0±0.2°, and 26.7±0.2°,

[0083] Alternatively, at 8.9±0.2°, 10.0±0.2°, 20.1±0.2°, 9.6±0.2°, 16.8±0.2°, 15.3±0.2°, 31.0±0.2°, 26.7±0.2°, 11.4±0.2°, and 24.2±0.2°.

[0084] The characteristic X-ray diffraction peaks of sodium salt crystal form 1 of compound 1 are shown in Table 12.

[0085] In a further preferred embodiment of this disclosure, the X-ray powder diffraction pattern of sodium salt crystal form 1 of compound 1 is shown in Figure 4.

[0086] In a further preferred embodiment of this disclosure, the 2θ error between the positions of the top ten diffraction peaks with the highest relative peak intensities in the X-ray powder diffraction pattern of the sodium salt crystal form 1 of compound 1 and the corresponding diffraction peaks in Figure 4 is ±0.2° to ±0.5°, preferably ±0.2° to ±0.3°, and more preferably ±0.2°.

[0087] In a further preferred embodiment of this disclosure, the crystal form is sodium salt crystal form 2 of compound 1, wherein:

[0088] The powder X-ray diffraction pattern of sodium salt crystal form 2 of compound 1 has a characteristic peak at 2θ of 10.3 ± 0.2°, or at 2θ of 20.5 ± 0.2°, or at 2θ of 8.9 ± 0.2°, or at 2θ of 7.8 ± 0.2°, or at 2θ of 17.1 ± 0.2°, or at 2θ of 15.3 ± 0.2°, or at 2θ of 26.6 ± 0.2°, or at 2θ of 24.2 ± 0.2°, or at 2θ of 12.9 ± 0.2°, or at 2θ of 11.9 ± 0.2°; preferably, it includes any 2, 4, 6, 8, or 10 characteristic peaks.

[0089] In a further preferred embodiment of this disclosure, the powder X-ray diffraction pattern of sodium salt crystal form 2 of compound 1 has one or more characteristic peaks at 10.3±0.2°, 20.5±0.2°, 8.9±0.2°, and 7.8±0.2°; preferably, it includes 2-4 such peaks, more preferably 3-4 such peaks, and most preferably 4 such peaks; optionally, it may further include one or more characteristic peaks at 2θ of 17.1±0.2°, 15.3±0.2°, 26.6±0.2°, 24.2±0.2°, or 11.9±0.2°, preferably 2, 3, 4, or 5 such peaks; for example, the X-ray powder diffraction pattern of sodium salt crystal form 2 of compound 1 has diffraction peaks at the following positions with 2θ:

[0090] At 10.3±0.2°, 20.5±0.2° and 8.9±0.2°,

[0091] Alternatively, at 10.3±0.2°, 20.5±0.2°, 8.9±0.2°, and 7.8±0.2°,

[0092] Alternatively, at 10.3±0.2°, 20.5±0.2°, 8.9±0.2°, 7.8±0.2°, 17.1±0.2°, and 15.3±0.2°.

[0093] In a further preferred embodiment of this disclosure, the X-ray powder diffraction pattern of the sodium salt crystal form 2 of compound 1 has diffraction peaks at one or more of the following 2θ values: 10.3±0.2°, 20.5±0.2°, 8.9±0.2°, 7.8±0.2°, 17.1±0.2°, 15.3±0.2°, 26.6±0.2°, 24.2±0.2°, 12.9±0.2°, and 11.9±0.2°. Preferably, it includes diffraction peaks at any of the following selected locations: 4, 6, 8, or 10 locations. For example, the X-ray powder diffraction pattern of the sodium salt crystal form 2 of compound 1 has diffraction peaks at the following 2θ positions:

[0094] At angles of 10.3±0.2°, 20.5±0.2°, 8.9±0.2°, 7.8±0.2°, 17.1±0.2°, 15.3±0.2°, 26.6±0.2°, and 24.2±0.2°,

[0095] Alternatively, at 10.3±0.2°, 20.5±0.2°, 8.9±0.2°, 7.8±0.2°, 17.1±0.2°, 15.3±0.2°, 26.6±0.2°, 24.2±0.2°, 12.9±0.2°, and 11.9±0.2°.

[0096] The characteristic X-ray diffraction peaks of sodium salt crystal form 2 of compound 1 are shown in Table 13.

[0097] In a further preferred embodiment of this disclosure, the X-ray powder diffraction pattern of sodium salt crystal form 2 of compound 1 is shown in Figure 5.

[0098] In a further preferred embodiment of this disclosure, the 2θ error between the positions of the top ten diffraction peaks with the highest relative peak intensities in the X-ray powder diffraction pattern of the sodium salt crystal form 2 of compound 1 and the corresponding diffraction peaks in Figure 5 is ±0.2° to ±0.5°, preferably ±0.2° to ±0.3°, and more preferably ±0.2°.

[0099] In a further preferred embodiment of this disclosure, the crystal form is sodium salt crystal form 1 of compound 2, wherein:

[0100] The powder X-ray diffraction pattern of sodium salt crystal form 1 of compound 2 has a characteristic peak at 2θ = 10.4 ± 0.2°, or at 2θ = 5.3 ± 0.2°, or at 2θ = 9.0 ± 0.2°, or at 2θ = 7.7 ± 0.2°, or at 2θ = 20.5 ± 0.2°, or at 2θ = 17.4 ± 0.2°, or at 2θ = 24.9 ± 0.2°, or at 2θ = 15.0 ± 0.2°. The characteristic peak may be present at 2θ = 11.9 ± 0.2°, or at 2θ = 21.2 ± 0.2°, or at 2θ = 19.9 ± 0.2°, or at 2θ = 16.5 ± 0.2°, or at 2θ = 18.0 ± 0.2°, or at 2θ = 15.9 ± 0.2°, or at 2θ = 26.8 ± 0.2°; preferably, the characteristic peak may be present at any of 2, 4, 6, 8, 10, or 12 of these locations.

[0101] In a further preferred embodiment of this disclosure, the powder X-ray diffraction pattern of sodium salt crystal form 1 of compound 2 has one or more characteristic peaks at 10.4±0.2°, 5.3±0.2°, 9.0±0.2°, or 7.7±0.2°; preferably including 2-4 such peaks, more preferably including 3-4 such peaks, and most preferably including 4 such peaks; optionally, further, it may also include one or more characteristic peaks at 2θ of 20.5±0.2°, 17.4±0.2°, 24.9±0.2°, 15.0±0.2°, or 11.9±0.2°, preferably including 2, 3, 4, or 5 such peaks; for example, the X-ray powder diffraction pattern of sodium salt crystal form 1 of compound 2 has diffraction peaks at the following positions at 2θ:

[0102] At 10.4±0.2°, 5.3±0.2° and 9.0±0.2°,

[0103] Alternatively, at 10.4±0.2°, 5.3±0.2°, 9.0±0.2°, and 7.7±0.2°,

[0104] Alternatively, at 10.4±0.2°, 5.3±0.2°, 9.0±0.2°, 7.7±0.2°, and 20.5±0.2°.

[0105] In a further preferred embodiment of this disclosure, the X-ray powder diffraction pattern of sodium salt crystal form 1 of compound 2 has diffraction peaks at one or more of the following 2θ values: 10.4±0.2°, 5.3±0.2°, 9.0±0.2°, 7.7±0.2°, 20.5±0.2°, 17.4±0.2°, 24.9±0.2°, 15.0±0.2°, 11.9±0.2°, 21.2±0.2°, 19.9±0.2°, 16.5±0.2°, 18.0±0.2°, 15.9±0.2°, and 26.8±0.2°; preferably, it includes diffraction peaks at any of 4, 6, 8, or 10 of these values; for example, the X-ray powder diffraction pattern of sodium salt crystal form 1 of compound 2 has diffraction peaks at the following 2θ positions:

[0106] At angles of 10.4±0.2°, 5.3±0.2°, 9.0±0.2°, 7.7±0.2°, 20.5±0.2°, 17.4±0.2°, 24.9±0.2°, and 15.0±0.2°,

[0107] Alternatively, at 10.4±0.2°, 5.3±0.2°, 9.0±0.2°, 7.7±0.2°, 20.5±0.2°, 17.4±0.2°, 24.9±0.2°, 15.0±0.2°, and 11.9±0.2°.

[0108] The characteristic X-ray diffraction peaks of sodium salt crystal form 1 of compound 2 are shown in Table 14.

[0109] In a further preferred embodiment of this disclosure, the X-ray powder diffraction pattern of sodium salt crystal form 1 of compound 2 is shown in Figure 6.

[0110] In a further preferred embodiment of this disclosure, the 2θ error between the positions of the top ten diffraction peaks with the highest relative intensities in the X-ray powder diffraction pattern of the sodium salt crystal form 1 of compound 2 and the corresponding diffraction peaks in Figure 6 is ±0.2° to ±0.5°, preferably ±0.2° to ±0.3°, and more preferably ±0.2°.

[0111] In a further preferred embodiment of this disclosure, the crystal form is potassium salt crystal form 1 of compound 2, wherein:

[0112] The powder X-ray diffraction pattern of potassium salt crystal form 1 of compound 2 has a characteristic peak at 2θ = 9.8 ± 0.2°, or at 2θ = 19.5 ± 0.2°, or at 2θ = 26.7 ± 0.2°, or at 2θ = 13.0 ± 0.2°, or at 2θ = 15.6 ± 0.2°, or at 2θ = 21.2 ± 0.2°, or at 2θ = 24.3 ± 0.2°, or at 2θ = 8.6 ± 0.2°. The peak has a characteristic peak at 2θ = 22.7 ± 0.2°, or at 2θ = 20.7 ± 0.2°, or at 2θ = 16.3 ± 0.2°, or at 2θ = 24.6 ± 0.2°, or at 2θ = 29.4 ± 0.2°, or at 2θ = 30.3 ± 0.2°, or at 2θ = 15.0 ± 0.2°; preferably, it includes any 2, 4, 6, 8, 10 or 12 of these peaks.

[0113] In a further preferred embodiment of this disclosure, the powder X-ray diffraction pattern of potassium salt crystal form 1 of compound 2 has one or more characteristic peaks at 9.8±0.2°, 19.5±0.2°, 26.7±0.2°, or 13.0±0.2°; preferably including 2-4 such peaks, more preferably including 3-4 such peaks, and most preferably including 4 such peaks; optionally, further, it may also include one or more characteristic peaks at 2θ of 15.6±0.2°, 21.2±0.2°, 24.3±0.2°, 8.6±0.2°, or 22.7±0.2°, preferably including 2, 3, 4, or 5 such peaks; for example, the X-ray powder diffraction pattern of potassium salt crystal form 1 of compound 2 has diffraction peaks at the following positions at 2θ:

[0114] At 9.8±0.2°, 19.5±0.2° and 26.7±0.2°,

[0115] Alternatively, at 9.8±0.2°, 19.5±0.2°, 26.7±0.2°, and 13.0±0.2°,

[0116] Alternatively, at 9.8±0.2°, 19.5±0.2°, 26.7±0.2°, 13.0±0.2°, 15.6±0.2°, and 21.2±0.2°.

[0117] In a further preferred embodiment of this disclosure, the X-ray powder diffraction pattern of the potassium salt crystal form 1 of compound 2 has diffraction peaks at one or more of the following 2θ values: 9.8±0.2°, 19.5±0.2°, 26.7±0.2°, 13.0±0.2°, 15.6±0.2°, 21.2±0.2°, 24.3±0.2°, 8.6±0.2°, 22.7±0.2°, 20.7±0.2°, 16.3±0.2°, 24.6±0.2°, 29.4±0.2°, 30.3±0.2°, and 15.0±0.2°; preferably, it includes diffraction peaks at any of 4, 6, 8, or 10 of these values; for example, the X-ray powder diffraction pattern of the potassium salt crystal form 1 of compound 2 has diffraction peaks at the following 2θ positions:

[0118] At 9.8±0.2°, 19.5±0.2°, 26.7±0.2°, 13.0±0.2°, 15.6±0.2°, 21.2±0.2°, 24.3±0.2°, and 8.6±0.2°,

[0119] Alternatively, at 9.8±0.2°, 19.5±0.2°, 26.7±0.2°, 13.0±0.2°, 15.6±0.2°, 21.2±0.2°, 24.3±0.2°, 8.6±0.2°, 22.7±0.2°, and 20.7±0.2°.

[0120] The characteristic X-ray diffraction peaks of potassium salt crystal form 1 of compound 2 are shown in Table 15.

[0121] In a further preferred embodiment of this disclosure, the X-ray powder diffraction pattern of potassium salt crystal form 1 of compound 2 is shown in Figure 7.

[0122] In a further preferred embodiment of this disclosure, the 2θ error between the positions of the top ten diffraction peaks with the highest relative intensities in the X-ray powder diffraction pattern of the potassium salt crystal form 1 of compound 2 and the corresponding diffraction peaks in Figure 7 is ±0.2° to ±0.5°, preferably ±0.2° to ±0.3°, and more preferably ±0.2°.

[0123] This disclosure also provides a method for preparing the acid salt or basic salt crystal form of the compound shown in formula (I), specifically comprising the following steps:

[0124] 1) Weigh an appropriate amount of compound (I) and dissolve it in a good solvent;

[0125] 2) Weigh an appropriate amount of the counter-ion base and dissolve it in an organic solvent; the amount of the counter-ion base is preferably 1.2 equivalents;

[0126] 3) Combine the two solutions and stir to precipitate.

[0127] 4) Dry to obtain the target product;

[0128] in:

[0129] The beneficial solvent is selected from methanol, ethyl acetate, acetone, tetrahydrofuran, isopropanol, or 2-butanone; preferably ethyl acetate or tetrahydrofuran.

[0130] The organic solvent is selected from methanol, ethanol, ethyl acetate, dichloromethane, acetone, acetonitrile, tetrahydrofuran, 2-butanone, 3-pentanone, tert-butanol, or N,N-dimethylformamide; ethanol is preferred; the above-mentioned benign solvents and organic solutions must be miscible when used.

[0131] The counterion base is an organic or inorganic base. The organic base is selected from sodium methoxide, potassium ethoxide, trimethylamine, diethylamine, triethylamine, triethanolamine, pyridine, piperidine, morpholine, diisopropylethylamine, diisopropylaminolithium, diethylaminolithium, bis(trimethylsilyl)aminolithium, potassium acetate, sodium acetate, isopropylcyclohexylaminolithium, or mixtures thereof. The inorganic base is selected from potassium phosphate, potassium phosphate trihydrate, potassium phosphate dihydrate, potassium phosphate monohydrate, sodium bicarbonate, potassium bicarbonate, sodium carbonate, cesium carbonate, potassium hydroxide, sodium hydroxide, potassium hydride, sodium hydride, lithium hydroxide, or mixtures thereof. Preferred bases are sodium methoxide or potassium hydroxide.

[0132] This disclosure also provides a method for preparing the sodium salt crystal form of the compound shown in formula (I), specifically comprising the following steps:

[0133] 1) Weigh an appropriate amount of the sodium salt of the compound and suspend it in a poor solvent. The preferred suspension density is 50-200 mg / mL.

[0134] 2) The suspension obtained above is stirred at a certain temperature for a certain time, preferably 0-50℃, and the time is preferably 1-10 days;

[0135] 3) The above suspension was rapidly centrifuged to remove the supernatant. The remaining solid was placed in a vacuum drying oven at 40°C and dried to constant weight to obtain the target product.

[0136] in:

[0137] The undesirable solvent is selected from methyl tert-butyl ether, isopropyl ether, diethyl ether, n-hexane, n-heptane, cyclohexane, n-pentane, water, and a mixture of water and the aforementioned benign solvents; preferably methyl tert-butyl ether, isopropyl ether, and n-heptane.

[0138] This disclosure also provides a method for preparing the acid salt or basic salt crystal form of the compound shown in formula (I), specifically comprising the following steps:

[0139] 1) Weigh an appropriate amount of compound (I) and dissolve it in a good solvent;

[0140] 2) Weigh an appropriate amount of the counterionic acid and dissolve it in an organic solvent;

[0141] 3) Combine the two solutions and stir to precipitate.

[0142] 4) Dry to obtain the target product;

[0143] in:

[0144] The beneficial solvent is selected from methanol, ethyl acetate, acetone, tetrahydrofuran, isopropanol, or 2-butanone; tetrahydrofuran is preferred.

[0145] The organic solvent is selected from methanol, ethanol, ethyl acetate, dichloromethane, acetone, acetonitrile, tetrahydrofuran, 2-butanone, 3-pentanone, tert-butanol, or N,N-dimethylformamide; ethanol is preferred; the above-mentioned benign solvents and organic solutions must be miscible when used.

[0146] The aforementioned counterionic acid is selected from inorganic or organic acids, wherein the inorganic acid is selected from hydrochloric acid, sulfuric acid, nitric acid, hydrobromic acid, hydrofluoric acid, hydroiodic acid, or phosphoric acid; and the organic acid is selected from 2,5-dihydroxybenzoic acid, 1-hydroxy-2-naphtholic acid, acetic acid, dichloroacetic acid, trichloroacetic acid, acetoxyxamic acid, adipic acid, benzenesulfonic acid, 4-chlorobenzenesulfonic acid, benzoic acid, 4-acetaminobenzoic acid, 4-aminobenzoic acid, decanoic acid, hexanoic acid, caprylic acid, cinnamic acid, citric acid, cyclohexanesulfonic acid, camphorsulfonic acid, aspartic acid, camphoric acid, gluconic acid, glucuronic acid, glutamic acid, isoascorbic acid, lactic acid, malic acid, mandelic acid, pyroglutamic acid, tartaric acid, etc. Dodecyl sulfuric acid, dibenzoyl tartaric acid, ethane-1,2-disulfonic acid, ethanesulfonic acid, formic acid, fumaric acid, galactobionic acid, gentian acid, glutaric acid, 2-ketoglutaric acid, glycolic acid, hippuric acid, hydroxyethyl sulfonic acid, lactobionic acid, ascorbic acid, aspartic acid, lauric acid, camphoric acid, maleic acid, malonic acid, methanesulfonic acid, 1,5-naphthalenedisulfonic acid, naphthalene-2-sulfonic acid, nicotinic acid, oleic acid, orotic acid, oxalic acid, palmitic acid, dihydroxynaphthalic acid, propionic acid, salicylic acid, 4-aminosalicylic acid, sebacic acid, stearic acid, succinic acid, thiocyanate, undecanoic acid, trifluoroacetic acid, benzenesulfonic acid, p-methylbenzenesulfonic acid, or L-malic acid; the counterionic acid is preferably sulfuric acid.

[0147] On one hand, this application provides a pharmaceutical composition containing a therapeutically effective amount of the crystal form of the compound described in this application, and one or more pharmaceutically acceptable carriers, diluents, or excipients.

[0148] Another object of this disclosure is to provide a method for preparing a pharmaceutical composition containing the compound described herein or a pharmaceutically acceptable salt thereof, comprising the step of mixing an acidic or basic salt form of the compound described herein with one or more pharmaceutically acceptable carriers, diluents, or excipients.

[0149] On the one hand, this application provides the use of the crystal form of the compound as described in this application or the pharmaceutical composition in the preparation of a medicament for treating cancer.

[0150] On the one hand, this application provides the use of the crystal form of the compound as described in this application or the pharmaceutical composition for the preparation of a medicament for treating a condition or disease that can be treated by WRN inhibition.

[0151] In a further preferred embodiment of this disclosure, the cancer or condition or disease is selected from the group consisting of microsatellite instability-high (MSI-H) or mismatch repair deficiency (dMMR), preferably, microsatellite instability-high (MSI-H) or mismatch repair deficiency (dMMR) selected from colorectal cancer, stomach cancer, prostate cancer, endometrial cancer, adrenocortical cancer, uterine cancer, cervical cancer, esophageal cancer, breast cancer, kidney cancer, and ovarian cancer. Attached Figure Description

[0152] Figure 1 shows the XRPD diagram of sulfate form A of compound 1.

[0153] Figure 2 shows the XRPD diagram of potassium salt A of compound 1.

[0154] Figure 3 shows the XRPD diagram of potassium salt B of compound 1.

[0155] Figure 4 shows the XRPD diagram of sodium salt crystal form 1 of compound 1.

[0156] Figure 5 shows the XRPD diagram of sodium salt crystal form 2 of compound 1.

[0157] Figure 6 shows the XRPD diagram of sodium salt crystal form 1 of compound 2.

[0158] Figure 7 shows the XRPD diagram of potassium salt form 1 of compound 2. Detailed Implementation

[0159] Unless otherwise stated, the terms used in the specification and claims have the following meanings.

[0160] In this disclosure, alkyl refers to a saturated aliphatic hydrocarbon group, which is a straight-chain or branched group containing 1 to 20 carbon atoms, preferably an alkyl group containing 1 to 8 carbon atoms, more preferably an alkyl group containing 1 to 6 carbon atoms, and most preferably an alkyl group containing 1 to 3 carbon atoms. Non-limiting examples include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, 1,1-dimethylpropyl, 1,2-dimethylpropyl, 2,2-dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, 1-ethyl-2-methylpropyl, 1,1,2-trimethylpropyl, 1,1-dimethylbutyl, 1,2-dimethylbutyl, 2,2-dimethylbutyl, 1,3-dimethylbutyl, 2-ethylbutyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, 2,3-dimethylbutyl, n-heptyl, 2-methylhexyl, 3-methylhexyl, 4-methylhexyl, 5-methylhexyl, 2, 3-Dimethylpentyl, 2,4-Dimethylpentyl, 2,2-Dimethylpentyl, 3,3-Dimethylpentyl, 2-Ethylpentyl, 3-Ethylpentyl, n-Octyl, 2,3-Dimethylhexyl, 2,4-Dimethylhexyl, 2,5-Dimethylhexyl, 2,2-Dimethylhexyl, 3,3-Dimethylhexyl, 4,4-Dimethylhexyl, 2-Ethylhexyl, 3-Ethylhexyl, 4-Ethylhexyl, 2-Methyl-2-Ethylpentyl, 2-Methyl-3-Ethylpentyl, n-Nonyl, 2-Methyl-2-Ethylhexyl, 2-Methyl-3-Ethylhexyl, 2,2-Diethylpentyl, n-Decyl, 3,3-Diethylhexyl, 2,2-Diethylhexyl, and their various branched isomers, etc.

[0161] The alkyl group can be substituted or unsubstituted. When substituted, the substituent can be substituted at any usable connection point. The substituent is preferably one or more of the following groups, independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxyl, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio, heterocycloalkylthio, oxo, carboxyl, or carboxylic acid ester group. The present disclosure preferably includes methyl, ethyl, isopropyl, tert-butyl, haloalkyl, deuteralkyl, alkoxy-substituted alkyl, and hydroxy-substituted alkyl. The hydroxy-substituted alkyl group may be 2-hydroxyisopropyl or 1-hydroxyethyl.

[0162] In this disclosure, a heterocyclic group refers to a saturated or partially unsaturated monocyclic or polycyclic heterocyclic group containing 3 to 20 ring atoms, wherein one or more ring atoms are selected from nitrogen, oxygen, or S(O). m(where m is an integer from 0 to 2) heteroatoms, but excluding the ring portions of -OO-, -OS-, or -SS-, with the remaining ring atoms being carbon. Preferably, it contains 3 to 12 ring atoms, of which 1 to 4 are heteroatoms; more preferably, it contains 3 to 10 ring atoms; even more preferably, it contains 3 to 8 ring atoms. Non-limiting examples of monocyclic heterocyclic groups include pyrrolyl, pyrrolidone, piperidin-2-keto, 3,4-dihydropyridine-2(1H)-keto, 4,5-dihydropyridazin-3(2H)-keto, azahexyl, oxacyclobutyl, oxacyclobutyl, oxacyclohexyl, imidazolyl, tetrahydrofuranyl, tetrahydrothiophene, dihydroimidazolyl, dihydrofuranyl, dihydropyrazolyl, dihydropyrrolyl, piperidinyl, piperazinyl, morpholinyl, thiomorpholinyl, homopiperazinyl, pyranyl, etc.; preferably pyrrolidone, piperidin-2-keto, 3,4-dihydropyridine-2(1H)-keto, 4,5-dihydropyridazine-3(2H)-keto, aziridine, oxazine, dihydropyrrolidone, tetrahydrofuranyl, pyrazolyl, morpholinyl, etc. Piperazinyl and pyranyl; more preferably dihydropyrrolyl, pyrrolidone, piperidin-2-keto, 3,4-dihydropyridin-2(1H)-keto, 4,5-dihydropyridazin-3(2H)-keto, aziridine, oxetane, oxetane, morpholinyl, piperidinyl, piperazinyl Pyranyl. Polycyclic heterocyclic groups include spirocyclic, fused-ring, and bridged-ring heterocyclic groups; wherein the spirocyclic, fused-ring, and bridged-ring heterocyclic groups involved are optionally connected to other groups by single bonds, or further cyclically linked to other cycloalkyl, heterocyclic, aryl, and heteroaryl groups by any two or more atoms on the ring.

[0163] The heterocyclic group can be optionally substituted or unsubstituted. When substituted, the substituent is preferably one or more of the following groups, independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxyl, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio, heterocycloalkylthio, oxo, carboxyl, or carboxylic acid ester group.

[0164] In this disclosure, aryl refers to a 6- to 14-membered all-carbon monocyclic or fused polycyclic (i.e., a ring sharing adjacent carbon atom pairs) group having a conjugated π-electron system, preferably 6- to 10-membered, more preferably 6- to 8-membered, such as phenyl and naphthyl, preferably phenyl. The aryl ring may be fused to a heteroaryl, heterocyclic, or cycloalkyl ring, wherein the ring connected to the parent structure is an aryl ring, and non-limiting examples include:

[0165] In this disclosure, alkoxy groups refer to -O- (alkyl) and -O- (unsubstituted cycloalkyl), wherein alkyl is defined as described above, preferably alkyl containing 1 to 8 carbon atoms, more preferably alkyl containing 1 to 6 carbon atoms, and most preferably alkyl containing 1 to 3 carbon atoms. Non-limiting examples of alkoxy groups include: methoxy, ethoxy, propoxy, butoxy, cyclopropoxy, cyclobutoxy, cyclopentoxy, and cyclohexoxy. Alkoxy groups may be optionally substituted or unsubstituted, and when substituted, the substituent is preferably one or more of the following groups independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxyl, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio, heterocycloalkylthio, carboxyl, or carboxylic acid ester group;

[0166] Non-limiting examples of alkoxy groups include propion-2-oxy, etc.

[0167] In this disclosure, alkyl halogroup refers to an alkyl group substituted with one or more halogens, wherein the alkyl group is as defined above. Non-limiting examples of alkyl halogroups include: trifluoromethyl, trifluoroethyl;

[0168] Non-limiting examples of haloalkyl groups also include: difluoromethyl, 1,1,2,2-tetrafluoroethyl, perfluoroethyl, etc.

[0169] In this disclosure, a haloalkoxy group refers to an alkoxy group substituted with one or more halogens, wherein the alkoxy group is as defined above;

[0170] The halogenated alkoxy group can be fully halogenated or partially halogenated, and the number of halogens can be 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, etc.; the halogen is preferably F, Cl, Br, I; for example, it can be trifluoromethoxy, difluoromethoxy, 1,1,2,2-tetrafluoroethoxy, perfluoroethoxy, etc.

[0171] In this disclosure, hydroxyalkyl means an alkyl group substituted with a hydroxyl group, wherein the alkyl group is as defined above.

[0172] In this disclosure, a haloalkyl means an alkyl group substituted with one or more halogens, wherein the alkyl group is as defined above.

[0173] In this disclosure, a haloalkoxy group refers to an alkoxy group substituted with one or more halogens, wherein the alkoxy group is as defined above.

[0174] "Hydroxy" refers to the -OH group.

[0175] "Halogen" refers to fluorine, chlorine, bromine, or iodine.

[0176] "Amino" refers to -NH2.

[0177] “Cyano” refers to -CN.

[0178] "Nitro" refers to -NO2.

[0179] "THF" refers to tetrahydrofuran.

[0180] “EtOAc” refers to ethyl acetate.

[0181] "DMSO" refers to dimethyl sulfoxide.

[0182] "LDA" refers to lithium diisopropylamine.

[0183] "DMAP" refers to 4-dimethylaminopyridine.

[0184] “EtMgBr” refers to ethyl magnesium bromide.

[0185] “HOSu” refers to N-hydroxysuccinimide.

[0186] “EDCl” refers to 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride.

[0187] "IPA" refers to isopropyl alcohol.

[0188] “MeOH” refers to methanol.

[0189] “EtOH” refers to ethanol.

[0190] "DMF" refers to N,N-dimethylformamide.

[0191] "DIPEA" refers to N,N-diisopropylethylamine.

[0192] “HEPES” refers to 4-hydroxyethylpiperazine ethanesulfonic acid.

[0193] The different terms such as "X is selected from A, B, or C", "X is selected from A, B, and C", "X is A, B, or C", and "X is A, B, and C" all express the same meaning, that is, X can be any one or more of A, B, and C.

[0194] "Optional" or "optionally" means that the event or situation described below may, but does not have to, occur, and the description includes the circumstances under which the event or situation may or may not occur.

[0195] "Substituted" refers to one or more hydrogen atoms in a group, preferably up to five, more preferably one to three hydrogen atoms, which are independently substituted by the corresponding number of substituents. It goes without saying that the substituents are only in their possible chemical positions, and those skilled in the art can determine (by experiment or theory) possible or impossible substitutions without much effort. For example, an amino or hydroxyl group with free hydrogen atoms may be unstable when bonded to a carbon atom with an unsaturated bond (such as an alkene).

[0196] "Stereoheterogeneity" includes three categories: geometric heterogeneity (cis-trans heterogeneity), optical heterogeneity, and conformational heterogeneity.

[0197] As used herein, the name of a compound is intended to cover all possible isomer forms, including stereoisomers of the compound (e.g., enantiomers, diastereomers, racemic mixtures or mixtures thereof and any mixture thereof).

[0198] All hydrogen atoms described in this disclosure can be replaced by their isotope deuterium, and any hydrogen atom in the compounds of the embodiments involved in this disclosure can also be replaced by a deuterium atom.

[0199] "Pharmaceutical composition" means a mixture containing one or more of the compounds described herein or their physiologically / pharmacologically acceptable salts or prodrugs, along with other chemical components, such as physiologically / pharmacologically acceptable carriers and excipients. The purpose of a pharmaceutical composition is to facilitate administration to a living organism, thereby promoting the absorption of the active ingredient and the exertion of its biological activity.

[0200] Those skilled in the art will understand that the composition comprises at least one pharmaceutically acceptable excipient selected from fillers, disintegrants, lubricants, humectants, binders, preservatives, sweeteners, flavoring agents, emulsifiers, suspending agents, flow aids or thickeners, pharmaceutical solvents, stabilizers, or antioxidants. This means that the composition may contain one or more functional ingredients, such as fillers and disintegrants, or it may refer to one or more substances containing a single functional ingredient, such as two different types of fillers.

[0201] X-ray powder diffraction (XRPD) refers to the experimentally observed diffraction pattern or parameters derived from it, characterized by peak positions (x-axis) and peak intensities (y-axis). Those skilled in the art will understand that experimental errors depend on instrument conditions, sample preparation, and sample purity. In particular, it is known to those skilled in the art that X-ray diffraction patterns typically change with instrument conditions, and appropriate error tolerances for XRPD are: 2θ ± 0.5°; 2θ ± 0.4°; 2θ ± 0.3°; 2θ ± 0.2°. It is particularly important to note that the relative intensities of X-ray diffraction patterns can also vary with experimental conditions, so the order of peak intensities cannot be considered the sole or decisive factor. Furthermore, the influence of experimental factors such as sample height can cause an overall shift in peak angles, which is generally permissible. Therefore, those skilled in the art will understand that any crystal form with characteristic peaks identical or similar to those of the patterns disclosed herein falls within the scope of this disclosure.

[0202] "TGA" refers to thermogravimetric analysis (TGA) experiments.

[0203] "DSC" refers to the Differential Scanning Calorimetry (DSC) experiment.

[0204] "HPLC" refers to High Performance Liquid Chromatography (HPLC) experiments.

[0205] "PK" refers to pharmacokinetic (PK) experiments.

[0206] "KF" refers to the Karl Fischer moisture determination (KF) experiment.

[0207] The present disclosure is further described below with reference to embodiments, but these embodiments are not intended to limit the scope of the present disclosure.

[0208] Preparation of compounds

[0209] The following examples are used to explain this disclosure, but should not be considered as limiting the scope of this disclosure. If specific conditions for the experimental methods are not specifically described in the examples of this disclosure, they are generally based on the conventional or recommended conditions of the raw material and product manufacturers. Reagents not specifically named are commercially available conventional reagents.

[0210] Preparation Example 1

[0211] (R,E)-N-[2-chloro-4-(trifluoromethyl)phenyl]-2-[2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-[[1-(5-hydroxy-6-methylpyrimidin-4-carbonyl)-5-methylpyrrolidine-3-ylidene]methyl]-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl]acetamide, compound 1

[0212] Step 1: (R)-2-methyl-4-((4,4,5,5-tetramethyl-1,3,2-dioxaborane-2-yl)methylene)pyrrolidine-1-carboxylic acid tert-butyl ester, 1-2

[0213] n-BuLi (2.5 M, 19.4 mL) was added to a solution of 2,2,6,6-tetramethylpiperidine (6.85 g, 48.5 mmol) in tetrahydrofuran (THF, 100 mL), and stirred at -30 °C for 0.5 h. After cooling to -78 °C, a solution of 4,4,5,5-tetramethyl-2-[(4,4,5,5-tetramethyl-1,3,2-dioxaborane-2-yl)methyl]-1,3,2-dioxaborane (10 g, 37.3 mmol) in THF (5 mL) was added and stirred for 0.5 h. Then, a solution of 1-1 (8.3 g, 44.8 mmol) in THF (50 mL) was added dropwise to the mixture, and the mixture was gradually heated to room temperature and stirred for 3 h. The mixture was then added to an aqueous solution of NH4Cl and stirred for 1 h. The mixture was filtered and concentrated under reduced pressure to remove THF. The residue was diluted with ethyl acetate (EA) and water, extracted once with EA, the organic layer was washed with brine, dried over Na₂SO₄ and concentrated under reduced pressure. The crude product was purified by column chromatography to give 1–2 (8.6 g, yield 74.53%, Z / E mixture, Z / E ratio approximately 75 / 25). Mass calc C 16 H 28 BNO4 for 309.21 found:310.26(M+H) + ESI.

[0214] Step 2: (R,E)-2-methyl-4-((4,4,5,5-tetramethyl-1,3,2-dioxaborane-2-yl)methylene)pyrrolidine-1-carboxylic acid tert-butyl ester, 1-2A

[0215] 1-2A was obtained by supercritical fluid chromatography (SFC) with a yield of 2 grams and a yield of 23.26%.

[0216] Preparative separation method: Instrument: WATERS150 preparative SFC (SFC-26); Column: ChiralPak AD, 250×30mm inner diameter, 10 μm; Mobile phase: A is carbon dioxide, B is ethanol, gradient: B 8%; Flow rate: 120 mL / min; Back pressure: 100 bar; Column temperature: 38℃; Wavelength: 220 nm.

[0217] Step 3: (R,E)-4-((4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)methylene)-2-methylpyrrolidine-1-carboxylic acid tert-butyl ester, 1-3

[0218] Int-2 (500 mg, 0.89 mmol) was suspended in 1,4-dioxane (15 mL), and H2O (1 mL), K3PO4 (567.81 mg, 2.67 mmol), and 1-2A (288.22 mg, 0.89 mmol) were added. The mixture was then degassed under argon for 10 min, followed by the addition of Pd(dppf)Cl2·DCM (36.14 mg, 44.58 μmol), and stirred overnight at 85 °C. The mixture was diluted with ethyl acetate (EA) and water, the organic layer was washed with brine, dried over Na2SO4, and concentrated under reduced pressure. The residue was purified by column chromatography to give 1-3 (260 mg, 43.06% yield).

[0219] Mass calc C 32 H 36 ClF3N6O5 for 676.24,679.24 found:577.29,579.3(M-Boc+H) + ESI.

[0220] Step 4: (R,E)-N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-((5-methylpyrrolidine-3-ylidene)methyl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide, 1-4

[0221] 1-3 (80 mg, 0.05 mmol) was suspended in dichloromethane (DCM, 2 mL), and trifluoroacetic acid (TFA, 0.5 mL) was added. The mixture was stirred overnight at room temperature. The mixture was washed with an aqueous sodium bicarbonate solution, dried over Na₂SO₄, and concentrated under reduced pressure. The crude product (RM) was concentrated under reduced pressure and used without purification.

[0222] Mass calc C 27 H 28 ClF3N6O3 for 576.19,578.18 found:577.3,579.4(M+H) + ESI.

[0223] Step 5: (R,E)-N-[2-chloro-4-(trifluoromethyl)phenyl]-2-[2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-[[1-(5-hydroxy-6-methylpyrimidin-4-carbonyl)-5-methylpyrrolidine-3-ylidene]methyl]-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl]acetamide, 1

[0224] 5-Hydroxy-6-methylpyrimidine-4-carboxylic acid (9.35 mg, 0.06 mmol), HOBT (16.39 mg, 0.12 mmol), and EDCI (23.26 mg, 0.12 mmol) were added to 1 mL of dichloromethane (DCM), followed by the addition of pyridine (14.39 mg, 0.18 mmol) at 0 °C. After stirring at 0 °C for 0.5 hours, 1-4 (35 mg, 0.06 mmol) was added and the mixture was stirred at room temperature for 2 hours. The crude product (RM) was purified by reversed-phase column chromatography (C18 column) to give product 1 (4.88 mg, yield 11.28%).

[0225] Mass calc C 33 H 32 ClF3N8O5 for 712.21,714.21found:711.32,713.30(MH) - ESI.

[0226] 1 H NMR(400MHz,DMSO-d6)δ:11.60(br,1H),10.40(br,1H),8.64-8.61(m,1H),8.10-8 .06(m,1H),7.96(s,1H),7.74-7.72(m,1H),6.87-6.84(m,1H),6.31-6.25(m,1H), 5.40-5.34(m,2H),4.80-4.59(m,2H),4.28-4.27(m,3H),3.84-3.79(m,2H),3.08- 2.99(m,1H),2.87-2.73(m,2H),2.55-2.54(m,3H),2.40(s,3H),1.23-1.08(m,6H).

[0227] Preparation Example 2

[0228] (E)-N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-((1-(5-hydroxy-6-methylpyrimidin-4-carbonyl)-5,5-dimethylpyrrolidine-3-ylidene)methyl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide, compound 2

[0229] Step 1: 2,2-Dimethyl-4-oxo-pyrrolidine-1-carboxylic acid tert-butyl ester, 2-2

[0230] 2-1 (5 g, 23.22 mmol) and DMP (14.78 g, 34.84 mmol) were mixed in dichloromethane (DCM, 10 mL) and stirred overnight at room temperature. The mixture was quenched with water, dried over Na2SO4, filtered, and concentrated under reduced pressure to obtain a residue. The residue was purified by column chromatography to give 2-2 (4.1 g, 19.22 mmol, yield 82.78%).

[0231] Detection was performed by thin-layer chromatography (TLC) 2-2.

[0232] Step 2: 2-(3-(allyloxy)but-1-en-2-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborane, 2-3

[0233] 2,2,6,6-Tetramethylpiperidine (3.18 g, 22.51 mmol) was dissolved in tetrahydrofuran (THF, 15 mL). After cooling to -30 °C, n-butyllithium (n-BuLi, 1.1 mL) was added and stirred for 30 min. After cooling to -78 °C, a solution of bis(4,4,5,5-tetramethyl-1,3,2-dioxaboron-2-yl)methane (6.03 g, 22.51 mmol) in tetrahydrofuran (THF, 15 mL) was added and stirred for 30 min. Then, a solution of 2-3 (4 g, 18.76 mmol) in tetrahydrofuran (THF, 15 mL) was added dropwise to the mixture and stirred overnight at room temperature. The mixture was quenched to 0 °C with aqueous NH4Cl solution and stirred for 30 min. After the reaction, the mixture was filtered and concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography to give 2-3 (5.5 g, yield 86.95%, Z / E mixture, Z / E ratio approximately 65 / 35). 2-3 was detected by thin-layer chromatography (TLC).

[0234] Step 3: (E)-2,2-dimethyl-4-((4,4,5,5-tetramethyl-1,3,2-dioxaborane-2-yl)methylene)pyrrolidine-1-carboxylic acid tert-butyl ester, 2-3A

[0235] 2-3A was obtained by supercritical fluid chromatography (SFC) with a yield of 1.5 g and a yield of 32.73%.

[0236] Preparative separation method: Instrument: MGⅡ preparative SFC (SFC-14); Column: ChiralPak IC, 250×30 mm inner diameter, 5 μm; Mobile phase: A is carbon dioxide, B is ethanol; Gradient: B 10%; Flow rate: 120 mL / min; Back pressure: 100 bar; Column temperature: 38℃; Wavelength: 220 nm; Cycle time: approximately 4 min.

[0237] Step 4: (E)-4-((4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)ylide)-2,2-dimethylpyrrolidine-1-carboxylic acid tert-butyl ester, 2-4

[0238] 2-[6-bromo-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4-yl]-N-[2-chloro-4-(trifluoromethyl)phenyl]acetamide (1 g, 1.78 mmol), (4E)-2,2-dimethyl-4-[(4,4,5,5-tetramethyl-1,3,2-dioxaborane-2-yl) [Methyl]pyrrolidine-1-carboxylic acid tert-butyl ester (721.73 mg, 2.14 mmol), K3PO4 (1.14 g, 5.35 mmol), and Pd(dppf)Cl2.DCM (72.28 mg, 89.17 μmol) were dissolved in 1,4-dioxane (20 mL) and H2O (2 mL), degassed three times with N2 gas, and then reacted under N2 atmosphere at 85 °C with stirring. The mixture was concentrated under reduced pressure to obtain a residue. The residue was purified by reversed-phase column chromatography (C18 column) to give products 2-4.

[0239] Mass calc C 33 H 38 O5N6ClF3 for 690.25,found:591.32(M-Boc+H) + ESI.

[0240] Step 5: (E)-N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-6-((5,5-dimethylpyrrolidone-3-ylidene)methyl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide, 2-5

[0241] Trifluoroacetic acid (TFA, 28.95 mg, 253.93 μmol, 0.5 mL) was added to a solution of 2-4 (390 mg, 253.93 μmol) in dichloromethane (DCM, 2 mL), and the mixture was stirred at room temperature for 3 hours. The mixture was extracted twice with an aqueous sodium bicarbonate solution, washed with brine, dried over Na₂SO₄, and concentrated under reduced pressure to obtain the residue. The crude product was used directly in the next reaction without purification.

[0242] Mass calc C 28 H 30O3N6ClF3 for 590.20,found:591.31(M+H) + ESI.

[0243] Step 6: (E)-N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-((1-(5-hydroxy-6-methylpyrimidin-4-carbonyl)-5,5-dimethylpyrrolidine-3-ylidene)methyl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide, compound 2

[0244] Pyridine (Py, 40.15 mg, 507.59 μmol, 40.89 μL) was added to a solution of 5-hydroxy-6-methylpyrimidine-4-carboxylic acid (39.12 mg, 253.80 μmol), HOBT (45.72 mg, 338.40 μmol), and EDCI (64.87 mg, 338.40 μmol) in dichloromethane (DCM, 10.06 mL) at 0 °C. The resulting solution was stirred at 0 °C for 15 min. Then, 2-5 (100.00 mg, 169.20 μmol) was added, and the resulting solution was stirred at room temperature for 18 h. The mixture was concentrated under reduced pressure and purified by reversed-phase column chromatography (C18 column) to give product compound 2 (50 mg, yield 40.64%).

[0245] Mass calc C 34 H 34 O5N8ClF3 for 726.23,found:725.20(MH) - ESI.

[0246] 1 H NMR(400MHz,DMSO-d6)δ:10.901(br,1H),10.390(br,1H),8.614(s,1H),8. 079-8.058(m,1H),7.979-7.976(m,1H),7.742-7.716(m,1H),6.851(s,1H) ,6.179(s,1H),5.350(s,2H),4.414-4.266(m,4H),3.831-3.804(m,2H),2. 776-2.673(m,4H),2.464-2.389(m,5H),1.512(s,6H),1.145-1.127(m,3H).

[0247] Biological Experiment

[0248] WRN helicase assay

[0249] To evaluate the inhibitory properties of the disclosed compounds on WRN helicase activity, a fluorescence assay using branched DNA was established. In the helicase assay, internally produced His-WRN was used. 527-1072 Fluorescent forked DNA prepared by annealing with OLIGOA-BHQ2 (TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTCGTACCCGATGTGTTCGTTC-BHQ2) and OLIGOA-TAMRA (TAMRA-GAACGAACACATCGGGTACGTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTT was used as substrate. The test compounds were prepared into 2.5-fold serial dilutions at 7 gradient points using DMSO, and then 1.5 μl of each concentration was added to 48.5 μl of assay buffer (25 mM Tris-HCl (pH 8.0), 50 mM NaCl, 2 mM MgCl2, 1 mM DTT, 0.05% Tween-20, and 2.5 μg / ml BSA) for further dilution. 5 μl of 45 nM WRN protein and 5 μl of each diluted compound solution were transferred to a 384-well assay plate and pre-incubated at room temperature for 30 min. Triples of each compound concentration were added to the plate. Controls were included in the assay to validate the assay. Instead of the compound solutions, assay buffer containing 3% DMSO was added to the positive control (no inhibition), and no protein was added to the negative control (maximum inhibition). The forked DNA and ATP were diluted to 300 nM and 6 mM, respectively, with assay buffer. To initiate the reaction, 5 μl of dsDNA and ATP solution were added to the assay plate. After incubation at room temperature for 60 minutes, the plate was transferred to a Victor Nivo multimode plate reader (Perkin Elmer, Waltham, MA) and fluorescence output was monitored. Dose-response curves were plotted using a GraphPad Prism 9 (GraphPad Software, San Diego, CA) to determine the IC50s.

[0250] WRN ATPase assay

[0251] To evaluate the inhibitory properties of the disclosed compounds against DNA-dependent WRN ATPase activity, an ATPase assay was established using the ADP-Glo ​​assay kit (Promega, Madison, WI). In the ATPase assay, the same WRN protein and DNA substrate as described in the helicase assay were used. Serial dilutions of the compounds in DMSO were the same as in the helicase assay. In further dilutions, 2 μl of each concentration was transferred to 48 μl of assay buffer. 5 μl of 100 nM WRN protein and 2.5 μl of each diluted compound solution were pre-incubated in a 384-well plate at room temperature for 30 min. All test wells were set in triplicate. Then, 2.5 μl of assay buffer containing 400 nM DNA substrate and 4 mM ATP was added to each well and incubated for 60 min. The assay included a protein-free control (maximum inhibition) and a compound-free control (no inhibition). To stop the reaction and consume excess ATP, 10 μl of reagent from the assay kit was added. TM The sample was incubated for 40 minutes. Then, 20 μl of ATP assay reagent was added. After incubation for 30 minutes, luminescence was measured using a Victor Nivo multimode plate reader (Perkin Elmer, Waltham, MA). The dose-response curve, spanning IC50s, was generated by a GraphPad Prism 9 (GraphPad Software, San Diego, CA).

[0252] Cell viability assay

[0253] The high microsatellite instability (MSI) cell line SW48 and the microsatellite stable MSS cell line SW480 were purchased from ATCC and cultured in DMEM GlutaMax medium (Gibco, 10564-011) consisting of penicillin-streptomycin (Gibco-15140122) and 10% fetal bovine serum (Gibco-10082-147) at 37°C in a humidified 5% CO2 incubator.

[0254] Cells were seeded at 3000 cells / well in white, clear-bottomed 384-well plates (PerkinElmer). The next day, twelve serially diluted (3-fold) aliquots of 10 mM DMSO stock solution were added to each replicate well. Blank wells contained cell-free culture medium, and DMSO control wells contained only cells and DMSO. After 72 hours of treatment, [the cells were then used...] Cell viability was analyzed using a 2.0 assay (Promega, G9243), and luminescence signals were detected on a VICTOR Nivo™ multimodal plate reader (PerkinElmer) after incubation at room temperature for 10 minutes. For data analysis, all data points were normalized by subtracting blank values, and the ratio of DMSO control values ​​was calculated. IC50 values ​​were calculated using a GraphPad Prism 9. The IC50 values ​​are shown in the table below.

[0255] Table 1

[0256] In vivo pharmacodynamic studies and pharmacokinetic / pharmacodynamic (PKPD) analysis

[0257] method

[0258] In vivo efficacy studies were conducted using 8- to 10-week-old female BALB / c nude mice (Shanghai Bikai Keyi Biotechnology Co., Ltd.). SW48 human colorectal cancer cells (CBP60018, COBIOER) were cultured in DMEM medium (11995-065, Gibco) supplemented with 10% FBS (A5669701, Gibco) at 37°C in a 5% CO2 incubator. A mixture of 3 million SW48 cells / Matrigel (356234, Corning) was subcutaneously injected into the right upper quadrant of the mice. Following inoculation, mice carrying tumors were monitored periodically, and when tumors reached an appropriate size, mice were randomly assigned to treatment groups (n = 6 or 7 per group).

[0259] The test compound was dissolved in a 10% 2-hydroxypropyl-β-cyclodextrin (HPBCD) solution. Mice were administered the test compound via oral gavage (PO) daily (QD) at a dose of 10 mpk, 20 mpk, or 40 mpk.

[0260] Mice were weighed twice weekly and tumor volume was measured. Tumor volume (mm) 3 According to the formula 0.5*D*d 2 Calculate, where D is the tumor length and d is the tumor width.

[0261] Tumor growth inhibition rate (TGI%) was calculated using the formula [1 - (△tumor volume in the treatment group / △tumor volume in the control group)] * 100; tumor regression rate was calculated using the formula - (△tumor volume in the treatment group / tumor volume in the treatment group on day 0) * 100, where △tumor volume represents the tumor volume on the measurement day minus the tumor volume on day 0. Drug efficacy was calculated using the formula (△tumor volume in the treatment group / △tumor volume in the control group) * 100. GraphPad Prism 10 was used to analyze tumor growth and weight change data.

[0262] Blood samples were collected in 1.5 ml EDTA-K2 tubes at various sampling time points following the last dose of the drug. Sampling time points were 0.25, 0.5, 1, 2, 4, 6, 8, and 24 hours. Blood samples were centrifuged at 8000 rpm for 5 minutes at room temperature. Plasma samples were transferred to pre-labeled tubes and stored at -80°C. For blood pharmacokinetic analysis, 5 μl of plasma sample was taken and 100 μl of internal standard (IS, glipizide, 50 ng / ml) in methanol was added. The mixture was vortexed for 1 minute and then centrifuged at 5228 g for 10 minutes. The supernatant was analyzed by LC-MS / MS-24 (Triple Quad 6500). Pharmacokinetic data were analyzed using a GraphPad Prism 10.

[0263] Tumors were collected, isolated, and processed using a dry ice rapid freezing method 4 hours or 24 hours after the last dose of the drug.

[0264] For tumor PK analysis, tumor samples were homogenized with 3 volumes (v / w) of PBS. 20 μl of sample was taken and 400 μl of internal standard (IS, glipizide, 50 ng / ml) in methanol was added. The mixture was vortexed for 1 min and then centrifuged at 5228 g for 10 min. The supernatant was analyzed by LC-MS / MS-24 (Triple Quad 6500). PK data were analyzed using a GraphPad Prism 10.

[0265] For tumor PD analysis, tumor tissue was prepared in 3-volume RIPA buffer containing phosphatase / protease inhibitors and homogenized on ice. The homogenized samples were repeatedly mixed and incubated on ice for 30 minutes. Centrifugation at 14,000g for 20 minutes at 4°C was performed to precipitate cell debris. The supernatant was transferred to fresh tubes for protein quantification using the Pierce BCA Protein Quantification Kit (#23225) and followed by Western blot analysis.

[0266] 30 μg of tumor protein lysis buffer was loaded onto a pre-prepared gel (4-12% Bis-Tris Midi Gels, Thermofisher, WG1403BOX), transferred to a polyvinylidene fluoride (PVDF) membrane or a pre-wetted nitrocellulose (NC) membrane, blocked with 5% skim milk diluted in Tris-buffered saline / Tween (TBST) for 1 hour, and then incubated overnight at 4°C with diluted primary antibody. Antibodies used for detecting protein targets, including WRN antibody (8H3, #4665), phosphorylated histone H2A.X Ser139 antibody (20E3#9718), phosphorylated Chk2 (Thr68) antibody (C13C1#2197), and p21 Waf1 / Cip1 antibody (12D1#2947), were all purchased from Cell Signaling Technology. After washing in 0.1% TBST for 5 minutes, repeated three times, the membrane was incubated with secondary antibody diluted in TBST containing 5% skim milk powder in a spinner at room temperature for 1 hour. Following membrane washing, target proteins were detected using a Tanon 5200 chemiluminescence imaging system and the ECL method. Loading controls were scanned using an Odyssey infrared imager. Data on target protein expression levels in tumors were analyzed using a GraphPad Prism 10.

[0267] result

[0268] The compounds disclosed herein exhibit good tumor-suppressive activity, safety, and PK properties.

[0269] Compound 2, administered orally daily at 10 mg / kg, showed a significant tumor regression rate of approximately 47% on day 28. Under the experimental conditions described above, mice treated with a 10 mpk dose showed a body weight change on day 28 within ±15%, more preferably within ±10%. The AUClast (hr*ng / ml) of the disclosed compounds in plasma or tumor (10 mpk, PO, last dose in the efficacy study) ranged from 1000 to 40000. Tables 3-5 show detailed results for representative compounds. No significant weight loss was observed in any of the treatment groups.

[0270] Table 2: Tumor Growth Inhibition Notes: i. * indicates tumor regression rate, meaning the tumor volume is less than the volume on day 0. ii. The formula for calculating the tumor growth inhibition rate (TGI%) is: [1 - (△tumor volume in the treatment group / △tumor volume in the control group)] * 100; iii. The formula for calculating the tumor regression rate is: -(△tumor volume in the treatment group / tumor volume in the treatment group on day 0) * 100, where △tumor volume represents the tumor volume on the measurement day minus the tumor volume on day 0.

[0271] Table 3: Weight Changes

[0272] 2. As shown in Table 4, the compounds disclosed herein have high exposure levels in plasma.

[0273] Table 4: PK Analysis of Efficacy Studies

[0274] Single-dose pharmacokinetics / pharmacodynamics (PKPD) of the SW48 xenograft model

[0275] Research Methods

[0276] The PKPD of the selected compounds was studied using the SW48 xenograft model.

[0277] SW48 human colorectal cancer cells (CBP60018, COBIOER) were cultured in DMEM medium (11995-065, Gibco) supplemented with 10% FBS (A5669701, Gibco) at 37°C in a 5% CO2 incubator.

[0278] BALB / c nude mice (female, 8-10 weeks old, Shanghai Bikai Keyi Biotechnology Co., Ltd.) were injected with a mixture of 3 million SW48 cells / Matrigel (356234, Corning). After inoculation, mice carrying tumors were monitored regularly, and when the tumors reached an appropriate size, the mice were randomly assigned to the treatment group (n=3 per group).

[0279] The test compound was dissolved in a 10% 2-hydroxypropyl-β-cyclodextrin (HPBCD) solution. Mice were administered the test compound via oral gavage (PO) daily (QD) at doses of 5 mpk, 10 mpk, 20 mpk, or 40 mpk.

[0280] Plasma PK sampling points were 0.25, 0.5, 1, 2, 4, 6, 8, 24, 48 hours and 72 hours after drug administration. Tumor PK / PD sampling points were 0.5, 2, 4, 8, 24, 48 hours and 72 hours after drug administration.

[0281] At the PK sampling site, blood samples were collected into 1.5 ml EDTA-K2 tubes. Blood samples were centrifuged at 8000 rpm for 5 minutes at room temperature. Plasma samples were transferred to pre-labeled tubes and stored at -80°C. Tumor samples were collected, separated, and rapidly frozen with dry ice. For blood PK analysis, 5 μl of plasma sample was taken and 100 μl of internal standard (IS, glipizide, 50 ng / ml) in methanol was added. The mixture was vortexed for 1 minute and then centrifuged at 5228 g for 10 minutes. The supernatant was analyzed by LC-MS / MS-24 (Triple Quad 6500). PK data were analyzed using a GraphPad Prism 10.

[0282] At the tumor sampling point, the tumor was collected, separated, and processed using a dry ice rapid freezing method.

[0283] For tumor pharmacokinetic (PK) analysis, tumor samples were homogenized with 3 volumes (v / w) of PBS. 20 μl of sample was taken and 400 μl of internal standard (IS, glipizide, 50 ng / ml) in methanol was added. The mixture was vortexed for 1 min and then centrifuged at 5228 g for 10 min. The supernatant was analyzed by LC-MS / MS-24 (Triple Quad 6500). When the plasma concentration was below the limit of detection (1 ng / ml), a PK plot was constructed using half the limit of detection. Data were analyzed using a GraphPad Prism 10.

[0284] For tumor PD analysis, tumor tissue was prepared in 3-volume RIPA buffer containing phosphatase / protease inhibitors and homogenized on ice. The homogenized samples were repeatedly mixed and incubated on ice for 30 minutes. Centrifugation at 14,000g for 20 minutes at 4°C was performed to precipitate cell debris. The supernatant was transferred to fresh tubes for protein quantification using the Pierce BCA Protein Quantification Kit (#23225) and followed by Western blot analysis.

[0285] 30 μg of tumor protein lysis buffer was loaded onto a pre-prepared gel (4-12% Bis-Tris Midi Gels, Thermofisher, WG1403BOX), transferred to a polyvinylidene fluoride (PVDF) membrane or a pre-wetted nitrocellulose (NC) membrane, blocked with 5% skim milk diluted in Tris-buffered saline / Tween (TBST) for 1 hour, and then incubated overnight at 4°C with diluted primary antibody. Antibodies used for detecting protein targets, including WRN antibody (8H3, #4665), phosphorylated histone H2A.X Ser139 antibody (20E3#9718), phosphorylated Chk2 (Thr68) antibody (C13C1#2197), and p21 Waf1 / Cip1 antibody (12D1#2947), were all purchased from Cell Signaling Technology. After washing in 0.1% TBST for 5 minutes, repeated three times, the membrane was incubated with secondary antibody diluted in TBST containing 5% skim milk powder in a spinner at room temperature for 1 hour. Following membrane washing, target proteins were detected using a Tanon 5200 chemiluminescence imaging system and the ECL method. Loading controls were scanned using an Odyssey infrared imager. Data on target protein expression levels in tumors were analyzed using a GraphPad Prism 10.

[0286] result

[0287] The compounds disclosed herein exhibit superior PK properties.

[0288] The AUClast (hr*ng / ml) (10mpk, PO, last dose in a pharmacodynamic study) of the disclosed compounds in plasma or tumor ranges from 5,000 to 60,000, more preferably from 10,000 to 50,000.

[0289] In vivo pharmacodynamic studies and pharmacokinetic / pharmacodynamic (PKPD) analysis

[0290] method

[0291] HCT-116 human colorectal cancer cells (JNO-H0125, Guangzhou Genio Biotechnology Co., Ltd.) were cultured in McCoy's 5A medium (12230-031, Gibco) supplemented with 10% FBS (A5669701, Gibco) at 37°C in a 5% CO2 incubator.

[0292] In vivo efficacy studies of HCT-116 were conducted using 8-10 week old female BALB / c nude mice (Shanghai Bikai Keyi Biotechnology Co., Ltd.). 4 million human colorectal cancer HCT-116 cells were subcutaneously injected into the right upper quadrant of the mice. After inoculation, mice carrying tumors were monitored periodically, and when the tumors reached an appropriate size, the mice were randomly assigned to treatment groups (n=7 per group).

[0293] The test compound was dissolved in a 10% 2-hydroxypropyl-β-cyclodextrin (HPBCD) solution. Mice were administered the test compound via oral gavage (PO) daily (QD) at a dose of 10 mpk, 20 mpk, or 40 mpk.

[0294] Mice were weighed twice weekly and tumor volume was measured. Tumor volume (mm) 3 According to the formula 0.5*D*d 2 Calculate, where D is the tumor length and d is the tumor width.

[0295] Tumor growth inhibition rate (TGI%) was calculated using the formula [1 - (△tumor volume in the treatment group / △tumor volume in the control group)] * 100; tumor regression rate was calculated using the formula - (△tumor volume in the treatment group / tumor volume in the treatment group on day 0) * 100, where △tumor volume represents the tumor volume on the measurement day minus the tumor volume on day 0. Drug efficacy was calculated using the formula (△tumor volume in the treatment group / △tumor volume in the control group) * 100. GraphPad Prism 10 was used to analyze tumor growth and weight change data.

[0296] Blood samples were collected in 1.5 ml EDTA-K2 tubes at various sampling time points following the last dose of the drug. Sampling time points were 0.25, 0.5, 1, 2, 4, 6, 8, and 24 hours. Blood samples were centrifuged at 8000 rpm for 5 minutes at room temperature. Plasma samples were transferred to pre-labeled tubes and stored at -80°C. For blood pharmacokinetic analysis, 5 μl of plasma sample was taken and 100 μl of internal standard (IS, glipizide, 50 ng / ml) in methanol was added. The mixture was vortexed for 1 minute and then centrifuged at 5228 g for 10 minutes. The supernatant was analyzed by LC-MS / MS-24 (Triple Quad 6500). Pharmacokinetic data were analyzed using a GraphPad Prism 10.

[0297] Tumors were collected, isolated, and processed using a dry ice rapid freezing method 4 hours or 24 hours after the last dose of the drug.

[0298] For tumor PK analysis, tumor samples were homogenized with 3 volumes (v / w) of PBS. 20 μl of sample was taken and 400 μl of internal standard (IS, glipizide, 50 ng / ml) in methanol was added. The mixture was vortexed for 1 min and then centrifuged at 5228 g for 10 min. The supernatant was analyzed by LC-MS / MS-24 (Triple Quad 6500). PK data were analyzed using a GraphPad Prism 10.

[0299] For tumor PD analysis, tumor tissue was prepared in 3-volume RIPA buffer containing phosphatase / protease inhibitors and homogenized on ice. The homogenized samples were repeatedly mixed and incubated on ice for 30 minutes. Centrifugation at 14,000g for 20 minutes at 4°C was performed to precipitate cell debris. The supernatant was transferred to fresh tubes for protein quantification using the Pierce BCA Protein Quantification Kit (#23225) and followed by Western blot analysis.

[0300] 30 μg of tumor protein lysis buffer was loaded onto a pre-prepared gel (4-12% Bis-Tris Midi Gels, Thermofisher, WG1403BOX), transferred to a polyvinylidene fluoride (PVDF) membrane or a pre-wetted nitrocellulose (NC) membrane, blocked with 5% skim milk diluted in Tris-buffered saline / Tween (TBST) for 1 hour, and then incubated overnight at 4°C with diluted primary antibody. Antibodies used for detecting protein targets, including WRN antibody (8H3, #4665), phosphorylated histone H2A.X Ser139 antibody (20E3#9718), phosphorylated Chk2 (Thr68) antibody (C13C1#2197), and p21 Waf1 / Cip1 antibody (12D1#2947), were all purchased from Cell Signaling Technology. After washing in 0.1% TBST for 5 minutes, repeated three times, the membrane was incubated with secondary antibody diluted in TBST containing 5% skim milk powder in a spinner at room temperature for 1 hour. Following membrane washing, target proteins were detected using a Tanon 5200 chemiluminescence imaging system and the ECL method. Loading controls were scanned using an Odyssey infrared imager. Data on target protein expression levels in tumors were analyzed using a GraphPad Prism 10.

[0301] result

[0302] The compounds disclosed herein possess good tumor-suppressive activity, safety profile, and pharmacokinetic properties.

[0303] Regarding tumor suppression, under the experimental conditions described above, the tumor growth inhibition rate (TGI) of the 10 mpk dose group exceeded 50% on day 28. More preferably, the TGI exceeded 80% on day 28. Even more preferably, the tumor regression rate reached 10-80% on day 28.

[0304] Under the above experimental conditions, the weight change of mice treated with a 10 mpk dose on day 28 was within ±15%, more preferably within ±10%.

[0305] The AUClast (hr*ng / ml) of the disclosed compounds in plasma or tumor (10mpk, orally, last dose in a pharmacodynamic study) ranges from 1,000 to 50,000, more preferably from 5,000 to 40,000.

[0306] Table 5-7 shows detailed results for representative compounds.

[0307] Compound 1 and Compound 2 were administered orally at doses of 3, 10, and 20 mpk daily. On day 28, the 20 mpk dose of Compound 1 showed a 50.36% reduction, while the 3, 10, and 20 mpk doses of Compound 2 showed 44.56%, 41.32%, and 59.75% reductions, respectively. No significant weight loss was observed in any of the treatment groups.

[0308] Table 5: Tumor Growth Inhibition Notes: i. * indicates tumor regression rate, meaning the tumor volume is less than the volume on day 0. ii. The formula for calculating the tumor growth inhibition rate (TGI%) is: [1 - (△tumor volume in the treatment group / △tumor volume in the control group)] * 100; iii. The formula for calculating the tumor regression rate is: -(△tumor volume in the treatment group / tumor volume in the treatment group on day 0) * 100, where △tumor volume represents the tumor volume on the measurement day minus the tumor volume on day 0.

[0309] Table 6: Weight Changes

[0310] Table 7: PK Analysis of Efficacy Studies

[0311] Crystal form study

[0312] 1. Experimental apparatus

[0313] 1.1 Some parameters of physicochemical testing instruments

[0314] 1.1.1 Physicochemical detection instrument parameters for the crystal forms of potassium salt and sulfate of compound 1

[0315] XRPD (X-ray Powder Diffraction) was used for analysis: measurements were performed using a BRUKER D8 X-ray diffractometer. Specific data collected included: Cu anode (40 kV, 40 mA), Cu-Kα1 rays. Kα2 rays Kβ rays Scanning range (2θ range): 3~40°, scanning step size 0.02, slit width (collimator) 1.0mm.

[0316] DSC stands for Differential Scanning Calorimetry: Measurements were performed using a METTLER TOLEDO DSC 3+ differential scanning calorimeter with a heating rate of 10℃ / min and a specific temperature range of 25-300℃. Nitrogen purging rate was 50mL / min.

[0317] TGA is thermogravimetric analysis: the test was performed using a METTLER TOLEDO TGA02 thermogravimetric analyzer, with a heating rate of 10℃ / min, a specific temperature range of 30-350℃, and a nitrogen purging rate of 50mL / min.

[0318] DVS stands for Dynamic Moisture Adsorption: The detection was performed using SMSDVS Advantage at 25℃, with humidity changes of 50%-95%-0%-95%-50% in 10% increments (5% in the final step) (see the graph for specific humidity ranges). The judgment criteria were Tmax 360 min and dm / dt not exceeding 0.002%.

[0319] 1.1.2 Physicochemical detection instrument parameters for sodium salt crystal form of compound 1, sodium salt crystal form 1 of compound 2, and potassium salt crystal form 1 of compound 2

[0320] The physicochemical detection instrument parameters for the sodium salt crystal form of compound 1, the sodium salt crystal form 1 of compound 2, and the potassium salt crystal form 1 of compound 2 are shown in Table 8.

[0321] Table 8

[0322] 2. Preparation of crystal form

[0323] 2.1 Preparation of sulfate crystal form A of compound 1

[0324] 8 mg of compound 1 was dissolved in 0.15 mL of tetrahydrofuran, and 6.17 μL of 2M sulfuric acid ethanol solution was added. The mixture was stirred at 5 °C to induce crystallization, filtered, and dried under vacuum to obtain a solid product. X-ray powder diffraction analysis showed that the product was sulfate crystal form A.

[0325] The XRPD spectrum is shown in Figure 1, and the positions of its characteristic peaks are shown in Table 9. The DSC spectrum shows endothermic peaks at 115.52℃, 228.96℃, and 234.28℃. The TGA spectrum shows a weight loss of 4.1% between 30℃ and 150℃. Ion detection results show a sulfate ion content of 12.31%.

[0326] Table 9

[0327] 2.2 Preparation of potassium salt crystal form A of compound 1

[0328] 200 mg of compound 1 was dissolved in 2.77 mL of tetrahydrofuran at room temperature, and 160 μL of 2M potassium hydroxide ethanol solution was added. The mixture was stirred at room temperature to induce crystallization, centrifuged, and dried under vacuum to obtain a solid product. X-ray powder diffraction analysis identified this product as potassium salt crystal form A. The XRPD spectrum is shown in Figure 2, and the characteristic peak positions are shown in Table 10. The DSC spectrum shows an endothermic peak at 148.21 °C. The TGA spectrum shows a weight loss of approximately 3.7% from 32 °C to 161 °C. Ion detection results showed a potassium ion content of 5.1%.

[0329] Table 10

[0330] 2.3 Preparation of potassium salt crystal form B of compound 1

[0331] 200 mg of compound 1 was dissolved in 3 mL of tetrahydrofuran, and 155 μL of 2 M potassium hydroxide ethanol solution was added. The mixture was stirred at 45 °C for 3.5 h, stirred at room temperature to induce crystallization, centrifuged, and dried under vacuum to obtain the solid product.

[0332] X-ray powder diffraction analysis identified the product as potassium salt crystal form B. The XRPD spectrum is shown in Figure 3, and the positions of its characteristic peaks are listed in Table 11. The DSC spectrum shows endothermic peaks at 66.81℃ and 129.50℃. The TGA spectrum shows a weight loss of 9.6% between 31℃ and 188℃.

[0333] Table 11

[0334] 2.4 Preparation of sodium salt crystal form 1 of compound 1

[0335] Weigh 200.34 mg of amorphous compound 1 into a 20 mL glass bottle, add 6 mL of ethyl acetate, and slowly add 118 μL of 15% sodium methoxide ethanol solution while stirring at 50 °C. Stir overnight at 50 °C to precipitate a white solid, filter, and vacuum dry at 40 °C for 8–10 hours. XRPD analysis showed that it was sodium salt crystal form 1.

[0336] Weigh 5g of amorphous compound 1 sample into a reaction vessel, add 150mL of ethyl acetate, and slowly add 600μL of 15% sodium methoxide ethanol solution while stirring at 50℃. Stir overnight at 50℃ to precipitate a white solid, filter, and dry under vacuum at 40℃ for 8-10 hours. XRPD analysis showed that it was sodium salt crystal form 1. The XRPD spectrum of sodium salt crystal form 1 of compound 1 is shown in Figure 4, and the positions of its characteristic peaks are shown in Table 12.

[0337] Table 12

[0338] 2.5 Preparation of sodium salt crystal form 2 of compound 1

[0339] Weigh 50 mg of sodium salt crystal form 1 sample and place it in a liquid chromatography vial. Add 0.5 mL of MTBE, suspend and stir at room temperature overnight, centrifuge, and then vacuum dry at 40 °C for 8–10 hours to obtain sodium salt crystal form 2.

[0340] Weigh 50 mg of sodium salt crystal form 1 sample and place it in a liquid chromatography vial. Add 0.5 mL of isopropyl ether, suspend and stir at room temperature overnight, centrifuge, and then vacuum dry at 40 °C for 8–10 hours to obtain sodium salt crystal form 2.

[0341] Weigh 50 mg of sodium salt crystal form 1 sample and place it in a liquid chromatography vial. Add 0.5 mL of n-heptane, suspend and stir at 50 °C overnight, centrifuge, and then vacuum dry at 40 °C for 8–10 hours to obtain sodium salt crystal form 2.

[0342] The XRPD spectrum of sodium salt form 2 of compound 1 is shown in Figure 5, and the positions of its characteristic peaks are shown in Table 13.

[0343] Table 13

[0344] 2.6 Preparation of sodium salt crystal form 1 of compound 2

[0345] Weigh 19.36 mg of amorphous compound 2 into a liquid chromatography vial, add 0.4 mL of ethyl acetate, stir at room temperature, slowly add 6.08 μL of 30% sodium methoxide ethanol solution, stir overnight at room temperature to precipitate a white solid, filter, and vacuum dry at 40 °C for 8–10 hours. XRPD detection showed that it was sodium salt crystal form 1 of compound 2.

[0346] Weigh 5g of amorphous compound 2 sample into a reaction vessel, add 100mL of ethyl acetate, and slowly add 200μL of 30% sodium methoxide ethanol solution while stirring at room temperature. Stir overnight at 50℃ to precipitate a white solid, filter, and vacuum dry at 40℃ for 8-10 hours. XRPD detection showed that it was sodium salt crystal form 1 of compound 2.

[0347] The XRPD spectrum of sodium salt of compound 2 (crystal form 1) is shown in Figure 6, and the positions of its characteristic peaks are shown in Table 14. The DSC spectrum shows endothermic peaks at 74.3℃ and 265.5℃. The TGA spectrum shows a weight loss of 11.70% between 35℃ and 150℃.

[0348] Table 14

[0349] 2.7 Preparation of potassium salt crystal form 1 of compound 2

[0350] Weigh 49.18 mg of amorphous compound 2 into a liquid chromatography vial, add 0.5 mL of tetrahydrofuran, stir at room temperature, slowly add 84.4 μL of 1 M potassium hydroxide ethanol solution, stir overnight at room temperature to precipitate a white solid, filter, vacuum dry at 40 °C for 8–10 hours, XRPD detection showed that it was sodium salt crystal form 1 of compound 2.

[0351] The XRPD spectrum of potassium salt form 1 of compound 2 is shown in Figure 7, and the positions of its characteristic peaks are shown in Table 15. The DSC spectrum shows endothermic peaks at 64.6℃ and 251.4℃. The TGA spectrum shows a weight loss of 6.01% from 35℃ to 150℃.

[0352] Table 15

[0353] Hygroscopicity of the 3 crystal forms

[0354] 3.2 Hygroscopicity of Compound 1 Sulfate Crystal Form A

[0355] DVS testing showed that under normal storage conditions (i.e., 25°C, 60% RH), the sample gained approximately 1.9% more weight due to moisture absorption; under accelerated testing conditions (i.e., 70% RH), the weight gain was approximately 2.4%; and under extreme conditions (90% RH), the weight gain was approximately 6.2%. Retesting of the crystal form after DVS testing showed no change in crystal form.

[0356] 3.1 Hygroscopicity of potassium salt form A of compound 1

[0357] DVS testing showed that under normal storage conditions (i.e., 25°C, 60% RH), the sample gained approximately 2.0% more weight due to moisture absorption; under accelerated testing conditions (i.e., 70% RH), the weight gain was approximately 2.5%; and under extreme conditions (90% RH), the weight gain was approximately 7.9%. Retesting of the crystal form after DVS testing showed no change in crystal form.

Claims

1. An acid salt or basic salt crystal form of a compound of general formula (I), wherein, The structure of the compound is shown below: in, X1 and X2 are independently selected from CH or N; R1 is selected from H, halogen, hydroxyl, C1-C6 alkyl, C1-C6 alkoxy, C1-C6 hydroxyalkyl, and C1-C6 haloalkyl; R2 is selected from H, halogen, hydroxyl, C1-C6 alkyl, C1-C6 alkoxy, C1-C6 hydroxyalkyl, and C1-C6 haloalkyl; R3 is selected from H, halogen, hydroxyl, C1-C6 alkyl, C1-C6 alkoxy, C1-C6 hydroxyalkyl, and C1-C6 haloalkyl; R4 is selected from H, halogen, hydroxyl, C1-C6 alkyl, C1-C6 alkoxy, C1-C6 hydroxyalkyl, and C1-C6 haloalkyl; R5 is selected from H, halogen, hydroxyl, C1-C6 alkyl, C1-C6 alkoxy, C1-C6 hydroxyalkyl, and C1-C6 haloalkyl; R6 is selected from H, halogen, hydroxyl, C1-C6 alkyl, C1-C6 alkoxy, C1-C6 hydroxyalkyl, and C1-C6 haloalkyl; R7 is selected from H, halogen, hydroxyl, C1-C6 alkyl, C1-C6 alkoxy, C1-C6 hydroxyalkyl, and C1-C6 haloalkyl; R8 is selected from H, halogen, hydroxyl, C1-C6 alkyl, C1-C6 alkoxy, C1-C6 hydroxyalkyl, and C1-C6 haloalkyl; R9 is selected from H, halogen, hydroxyl, C1-C6 alkyl, C1-C6 alkoxy, C1-C6 hydroxyalkyl, and C1-C6 haloalkyl; R 10 Selected from heterocyclic groups, wherein the heterocyclic group is a 5- or 6-membered fully saturated or partially unsaturated group, comprising a cyclic carbon atom and one or two cyclic heteroatoms selected from N, O and S respectively; The acid is an inorganic or organic acid, wherein the inorganic acid is selected from hydrochloric acid, sulfuric acid, nitric acid, hydrobromic acid, hydrofluoric acid, hydroiodic acid, or phosphoric acid; the organic acid is selected from 2,5-dihydroxybenzoic acid, 1-hydroxy-2-naphtholic acid, acetic acid, dichloroacetic acid, trichloroacetic acid, acetoxyxamic acid, adipic acid, benzenesulfonic acid, 4-chlorobenzenesulfonic acid, benzoic acid, 4-acetaminobenzoic acid, 4-aminobenzoic acid, decanoic acid, hexanoic acid, caprylic acid, cinnamic acid, citric acid, cyclohexanesulfonic acid, camphorsulfonic acid, aspartic acid, camphoric acid, gluconic acid, glucuronic acid, glutamic acid, isoascorbic acid, lactic acid, malic acid, mandelic acid, pyroglutamic acid, and alcohol. Sarcoic acid, dodecyl sulfuric acid, dibenzoyl tartaric acid, ethane-1,2-disulfonic acid, ethanesulfonic acid, formic acid, fumaric acid, galactobionic acid, gentian acid, glutaric acid, 2-ketoglutaric acid, glycolic acid, hippuric acid, hydroxyethyl sulfonic acid, lactobionic acid, ascorbic acid, aspartic acid, lauric acid, camphoric acid, maleic acid, malonic acid, methanesulfonic acid, 1,5-naphthalenedisulfonic acid, naphthalene-2-sulfonic acid, nicotinic acid, oleic acid, orotic acid, oxalic acid, palmitic acid, dihydroxynaphthalic acid, propionic acid, salicylic acid, 4-aminosalicylic acid, sebacic acid, stearic acid, succinic acid, thiocyanate, undecanoic acid, trifluoroacetic acid, benzenesulfonic acid, p-methylbenzenesulfonic acid, or L-malic acid; The base is an organic or inorganic base. The organic base is selected from sodium methoxide, potassium ethoxide, trimethylamine, diethylamine, triethylamine, triethanolamine, pyridine, piperidine, morpholine, diisopropylethylamine, diisopropylaminolithium, diethylaminolithium, bis(trimethylsilyl)aminolithium, potassium acetate, sodium acetate, isopropylcyclohexylaminolithium or mixtures thereof. The inorganic base is selected from potassium phosphate, potassium phosphate trihydrate, potassium phosphate dihydrate, potassium phosphate monohydrate, sodium bicarbonate, potassium bicarbonate, sodium carbonate, cesium carbonate, potassium hydroxide, sodium hydroxide, potassium hydride, sodium hydride, lithium hydroxide or mixtures thereof.

2. The acidic salt or basic salt crystal form of the compound according to claim 1, characterized in that, X1 and X2 are N; R1 is selected from C1-C3 alkyl groups; R2 is selected from F, Cl, Br, and I; R3 is selected from C 1-3 Halogenated alkyl groups, preferably CF3; R4 is selected from hydrogen; R5 is selected from OH; R6 is selected from C1-C3 alkyl groups; R7 is selected from hydrogen; R8 is selected from H or C1-C3 alkyl groups; R9 is selected from C1-C3 alkyl groups; R 10 Selected from 3. The acidic salt or basic salt crystal form of the compound according to claim 1, characterized in that, The compound is selected from the following compounds 1 or 2:

4. The acid salt or basic salt crystal form of the compound according to any one of claims 1-3, characterized in that, The salt crystal form can be a solvent-containing or solvent-free crystal form, wherein the solvent is selected from water, methanol, acetone, ethyl acetate, acetonitrile, ethanol, 88% acetone, 2-methyltetrahydrofuran, dichloromethane, 1,4-dioxane, benzene, toluene, isopropanol, n-butanol, isobutanol, N,N-dimethylformamide, N,N-dimethylacetamide, dimethyl sulfoxide, n-propanol, tert-butanol, 2-butanone, 3-pentanone, n-heptane, ethyl formate, isopropyl acetate, cyclohexane, methyl tert-butyl ether, or isopropyl ether; preferably, the number of solvents is 0.2-3, more preferably 0.2, 0.5, 1, 1.5, 2, 2.5, or 3, and most preferably 0.5, 1, 2, or 3; the crystal form is more preferably anhydrous.

5. The acid salt crystal form or basic salt crystal form according to any one of claims 1-3, characterized in that, The acidic salt crystal form is the sulfate crystal form, and the basic salt crystal form is the potassium salt crystal form or the sodium salt crystal form.

6. The acid salt crystalline form or base salt crystalline form of claim 5, characterized by, The compound is compound 1 or compound 2, and the acid salt crystal form or basic salt crystal form is selected from the sulfate crystal form A of compound 1, the potassium salt crystal form A of compound 1, the potassium salt crystal form B of compound 1, the sodium salt crystal form 1 of compound 1, the sodium salt crystal form 2 of compound 1, the sodium salt crystal form 1 of compound 2, and the potassium salt crystal form 1 of compound 2, wherein: The powder X-ray diffraction pattern of sulfate crystal form A of compound 1 has a characteristic peak at 2θ of 13.8 ± 0.2°, or at 2θ of 16.6 ± 0.2°, or at 2θ of 17.7 ± 0.2°, or at 2θ of 15.3 ± 0.2°, or at 2θ of 9.0 ± 0.2°, or at 2θ of 25.1 ± 0.2°, or at 2θ of 22.6 ± 0.2°, or at 2θ of 21. The characteristic peak is present at 0 ± 0.2°, or at 2θ = 20.2 ± 0.2°, or at 2θ = 23.5 ± 0.2°, or at 2θ = 9.9 ± 0.2°, or at 2θ = 11.1 ± 0.2°, or at 2θ = 27.8 ± 0.2°, or at 2θ = 14.6 ± 0.2°; preferably, the characteristic peak is present at any 2, 4, 6, 8, 10, or 12 of these locations. The powder X-ray diffraction pattern of potassium salt crystal form A of compound 1 has a characteristic peak at 2θ of 5.7 ± 0.2°, or at 2θ of 16.0 ± 0.2°, or at 2θ of 21.3 ± 0.2°, or at 2θ of 17.0 ± 0.2°, or at 2θ of 13.9 ± 0.2°, or at 2θ of 25.5 ± 0.2°, or at 2θ of 29.7 ± 0.2°, or at 2θ of 28.6 ± 0.2°. The peak has a characteristic peak at 2θ = 34.0 ± 0.2°, or at 2θ = 20.5 ± 0.2°, or at 2θ = 29.2 ± 0.2°, or at 2θ = 37.3 ± 0.2°, or at 2θ = 26.2 ± 0.2°, or at 2θ = 30.9 ± 0.2°, or at 2θ = 11.3 ± 0.2°; preferably, it includes any 2, 4, 6, 8, 10 or 12 of these peaks. The powder X-ray diffraction pattern of potassium salt crystal form B of compound 1 has a characteristic peak at 2θ of 5.2 ± 0.2°, or at 2θ of 7.4 ± 0.2°, or at 2θ of 14.5 ± 0.2°, or at 2θ of 16.1 ± 0.2°, or at 2θ of 18.7 ± 0.2°, or at 2θ of 21.8 ± 0.2°, or at 2θ of 11.7 ± 0.2°, or at 2θ of 10.0 ± 0.2°, or at 2θ of 26.0 ± 0.2°, or at 2θ of 20.4 ± 0.2°; preferably, it includes any 2, 4, 6, 8, or 10 characteristic peaks. The powder X-ray diffraction pattern of sodium salt crystal form 1 of compound 1 has a characteristic peak at 2θ = 8.9 ± 0.2°, or at 2θ = 10.0 ± 0.2°, or at 2θ = 20.1 ± 0.2°, or at 2θ = 9.6 ± 0.2°, or at 2θ = 16.8 ± 0.2°, or at 2θ = 15.3 ± 0.2°, or at 2θ = 31.0 ± 0.2°, or at 2θ = 26.7 ± 0.2°, or at 2θ = 11.4 ± 0.2°, or at 2θ = 24.2 ± 0.2°; preferably, it includes any 2, 4, 6, 8, or 10 characteristic peaks. The powder X-ray diffraction pattern of sodium salt crystal form 2 of compound 1 has a characteristic peak at 2θ of 10.3 ± 0.2°, or at 2θ of 20.5 ± 0.2°, or at 2θ of 8.9 ± 0.2°, or at 2θ of 7.8 ± 0.2°, or at 2θ of 17.1 ± 0.2°, or at 2θ of 15.3 ± 0.2°, or at 2θ of 26.6 ± 0.2°, or at 2θ of 24.2 ± 0.2°, or at 2θ of 12.9 ± 0.2°, or at 2θ of 11.9 ± 0.2°; preferably, it includes any 2, 4, 6, 8, or 10 characteristic peaks. The powder X-ray diffraction pattern of sodium salt crystal form 1 of compound 2 has a characteristic peak at 2θ = 10.4 ± 0.2°, or at 2θ = 5.3 ± 0.2°, or at 2θ = 9.0 ± 0.2°, or at 2θ = 7.7 ± 0.2°, or at 2θ = 20.5 ± 0.2°, or at 2θ = 17.4 ± 0.2°, or at 2θ = 24.9 ± 0.2°, or at 2θ = 15.0 ± 0.2°. The characteristic peak may be present at 2θ = 11.9 ± 0.2°, 2θ = 21.2 ± 0.2°, 2θ = 19.9 ± 0.2°, 2θ = 16.5 ± 0.2°, 2θ = 18.0 ± 0.2°, 2θ = 15.9 ± 0.2°, or 2θ = 26.8 ± 0.2°; preferably, the characteristic peak may be present at any of 2, 4, 6, 8, 10, or 12 of these locations. The powder X-ray diffraction pattern of potassium salt crystal form 1 of compound 2 has a characteristic peak at 2θ = 9.8 ± 0.2°, or at 2θ = 19.5 ± 0.2°, or at 2θ = 26.7 ± 0.2°, or at 2θ = 13.0 ± 0.2°, or at 2θ = 15.6 ± 0.2°, or at 2θ = 21.2 ± 0.2°, or at 2θ = 24.3 ± 0.2°, or at 2θ = 8.6 ± 0.2°. The peak has a characteristic peak at 2θ = 22.7 ± 0.2°, or at 2θ = 20.7 ± 0.2°, or at 2θ = 16.3 ± 0.2°, or at 2θ = 24.6 ± 0.2°, or at 2θ = 29.4 ± 0.2°, or at 2θ = 30.3 ± 0.2°, or at 2θ = 15.0 ± 0.2°; preferably, it includes any 2, 4, 6, 8, 10 or 12 of these peaks.

7. The acidic salt crystal form or basic salt crystal form according to claim 6, characterized in that, The powder X-ray diffraction pattern of sulfate crystal form A of compound 1 has one or more characteristic peaks at 13.8±0.2°, 16.6±0.2°, 17.7±0.2° or 15.3±0.2°; preferably including 2-4 of these peaks, more preferably including 3-4, and most preferably including 4. Optionally, further, it may also include one or more characteristic peaks at 2θ of 9.0±0.2°, 25.1±0.2°, 22.6±0.2°, 21.0±0.2°, 20.2±0.2°, preferably including 2, 3, 4 or 5 of these peaks; for example, the X-ray powder diffraction pattern of sulfate crystal form A of compound 1 has diffraction peaks at the following positions at 2θ: At 13.8±0.2°, 16.6±0.2° and 17.7±0.2°, Alternatively, at 13.8±0.2°, 16.6±0.2°, 17.7±0.2°, and 15.3±0.2°, Alternatively, at 13.8±0.2°, 16.6±0.2°, 17.7±0.2°, 15.3±0.2°, and 9.0±0.2°; The powder X-ray diffraction pattern of potassium salt crystal form A of compound 1 has one or more characteristic peaks at 5.7±0.2°, 16.0±0.2°, 21.3±0.2° or 17.0±0.2°; preferably including 2-4 of these peaks, more preferably including 3-4, and most preferably including 4. Optionally, it may further include one or more characteristic peaks at 2θ of 13.9±0.2°, 25.5±0.2°, 29.7±0.2°, 28.6±0.2°, or 34.0±0.2°, preferably including 2, 3, 4 or 5 of these peaks; for example, the X-ray powder diffraction pattern of potassium salt crystal form A of compound 1 has diffraction peaks at the following positions at 2θ: At 5.7±0.2°, 16.0±0.2°, and 21.3±0.2°, Alternatively, at 5.7±0.2°, 16.0±0.2°, 21.3±0.2°, and 17.0±0.2°, Alternatively, at 5.7±0.2°, 16.0±0.2°, 21.3±0.2°, 17.0±0.2°, 13.9±0.2°, and 25.5±0.2°; The powder X-ray diffraction pattern of potassium salt form B of compound 1 has one or more characteristic peaks at 5.2±0.2°, 7.4±0.2°, 14.5±0.2°, or 16.1±0.2°; preferably including 2-4 of these peaks, more preferably including 3-4, and most preferably including 4. Optionally, it may further include one or more characteristic peaks at 2θ of 18.7±0.2°, 21.8±0.2°, 11.7±0.2°, 10.0±0.2°, or 26.0±0.2°, preferably including 2, 3, 4, or 5 of these peaks; for example, the X-ray powder diffraction pattern of potassium salt form B of compound 1 has diffraction peaks at the following positions at 2θ: At 5.2±0.2°, 7.4±0.2°, and 14.5±0.2°, Alternatively, at 5.2±0.2°, 7.4±0.2°, 14.5±0.2°, and 16.1±0.2°, Alternatively, at 5.2±0.2°, 7.4±0.2°, 14.5±0.2°, 16.1±0.2°, 18.7±0.2°, and 21.8±0.2°; The powder X-ray diffraction pattern of sodium salt crystal form 1 of compound 1 has one or more characteristic peaks at 8.9±0.2°, 10.0±0.2°, 20.1±0.2°, and 9.6±0.2°; preferably including 2-4 of these peaks, more preferably including 3-4, and most preferably including 4. Optionally, further, it may also include one or more characteristic peaks at 2θ of 16.8±0.2°, 15.3±0.2°, 31.0±0.2°, 26.7±0.2°, or 11.4±0.2°, preferably including 2, 3, 4, or 5 of these peaks; for example, the X-ray powder diffraction pattern of sodium salt crystal form 1 of compound 1 has diffraction peaks at the following positions at 2θ: At 8.9±0.2°, 10.0±0.2°, and 20.1±0.2°, Alternatively, at 8.9±0.2°, 10.0±0.2°, 20.1±0.2°, and 9.6±0.2°, Alternatively, at 8.9±0.2°, 10.0±0.2°, 20.1±0.2°, 9.6±0.2°, 16.8±0.2°, and 15.3±0.2°; The powder X-ray diffraction pattern of sodium salt crystal form 2 of compound 1 has one or more characteristic peaks at 10.3±0.2°, 20.5±0.2°, 8.9±0.2°, and 7.8±0.2°; preferably including 2-4 of these peaks, more preferably including 3-4, and most preferably including 4. Optionally, further, it may also include one or more characteristic peaks at 2θ of 17.1±0.2°, 15.3±0.2°, 26.6±0.2°, 24.2±0.2°, or 11.9±0.2°, preferably including 2, 3, 4, or 5 of these peaks; for example, the X-ray powder diffraction pattern of sodium salt crystal form 2 of compound 1 has diffraction peaks at the following positions at 2θ: At 10.3±0.2°, 20.5±0.2° and 8.9±0.2°, Alternatively, at 10.3±0.2°, 20.5±0.2°, 8.9±0.2°, and 7.8±0.2°, Alternatively, at 10.3±0.2°, 20.5±0.2°, 8.9±0.2°, 7.8±0.2°, 17.1±0.2°, and 15.3±0.2°; The powder X-ray diffraction pattern of sodium salt crystal form 1 of compound 2 has one or more characteristic peaks at 10.4±0.2°, 5.3±0.2°, 9.0±0.2° or 7.7±0.2°; preferably including 2-4 of these peaks, more preferably including 3-4, and most preferably including 4. Optionally, it may further include one or more characteristic peaks at 2θ of 20.5±0.2°, 17.4±0.2°, 24.9±0.2°, 15.0±0.2°, or 11.9±0.2°, preferably including 2, 3, 4 or 5 of these peaks; for example, the X-ray powder diffraction pattern of sodium salt crystal form 1 of compound 2 has diffraction peaks at the following positions at 2θ: At 10.4±0.2°, 5.3±0.2° and 9.0±0.2°, Alternatively, at 10.4±0.2°, 5.3±0.2°, 9.0±0.2°, and 7.7±0.2°, Alternatively, at 10.4±0.2°, 5.3±0.2°, 9.0±0.2°, 7.7±0.2°, and 20.5±0.2°; The powder X-ray diffraction pattern of potassium salt crystal form 1 of compound 2 has one or more characteristic peaks at 9.8±0.2°, 19.5±0.2°, 26.7±0.2° or 13.0±0.2°; preferably including 2-4 of these peaks, more preferably including 3-4, and most preferably including 4. Optionally, further, it may also include one or more characteristic peaks at 2θ of 15.6±0.2°, 21.2±0.2°, 24.3±0.2°, 8.6±0.2°, or 22.7±0.2°, preferably including 2, 3, 4 or 5 of these peaks; for example, the X-ray powder diffraction pattern of potassium salt crystal form 1 of compound 2 has diffraction peaks at the following positions at 2θ: At 9.8±0.2°, 19.5±0.2° and 26.7±0.2°, Alternatively, at 9.8±0.2°, 19.5±0.2°, 26.7±0.2°, and 13.0±0.2°, Alternatively, at 9.8±0.2°, 19.5±0.2°, 26.7±0.2°, 13.0±0.2°, 15.6±0.2°, and 21.2±0.2°.

8. The acidic salt crystal form or the basic salt crystal form according to claim 6, characterized in that, The X-ray powder diffraction pattern of sulfate crystal form A of compound 1 has diffraction peaks at one or more of the following positions at 2θ: 13.8±0.2°, 16.6±0.2°, 17.7±0.2°, 15.3±0.2°, 9.0±0.2°, 25.1±0.2°, 22.6±0.2°, 21.0±0.2°, 20.2±0.2°, 23.5±0.2°, 9.9±0.2°, 11.1±0.2°, 27.8±0.2°, and 14.6±0.2°; preferably, it includes diffraction peaks at any of 4, 6, 8, or 10 of these positions; for example, the X-ray powder diffraction pattern of sulfate crystal form A of compound 1 has diffraction peaks at the following positions at 2θ: At 13.8±0.2°, 16.6±0.2°, 17.7±0.2°, 15.3±0.2°, 9.0±0.2°, 25.1±0.2°, 22.6±0.2°, and 21.0±0.2°, Alternatively, at 13.8±0.2°, 16.6±0.2°, 17.7±0.2°, 15.3±0.2°, 9.0±0.2°, 25.1±0.2°, 22.6±0.2°, 21.0±0.2°, 20.2±0.2°, and 23.5±0.2°, The powder X-ray diffraction pattern of potassium salt crystal form A of compound 1 has characteristic peaks at one or more of the following positions at 2θ: 5.7±0.2°, 16.0±0.2°, 21.3±0.2°, 17.0±0.2°, 13.9±0.2°, 25.5±0.2°, 29.7±0.2°, 28.6±0.2°, 34.0±0.2°, 20.5±0.2°, 29.2±0.2°, 37.3±0.2°, 26.2±0.2°, 30.9±0.2°, and 11.3±0.2°; preferably, diffraction peaks are found at any of 4, 6, 8, or 10 of these positions. For example, the X-ray powder diffraction pattern of potassium salt crystal form A has diffraction peaks at the following positions at 2θ: At 5.7±0.2°, 16.0±0.2°, 21.3±0.2°, 17.0±0.2°, 13.9±0.2°, 25.5±0.2°, 29.7±0.2°, and 28.6±0.2°, Alternatively, at 5.7±0.2°, 16.0±0.2°, 21.3±0.2°, 17.0±0.2°, 13.9±0.2°, 25.5±0.2°, 29.7±0.2°, 28.6±0.2°, 34.0±0.2°, and 20.5±0.2°; The powder X-ray diffraction pattern of potassium salt crystal form B of compound 1 has characteristic peaks at one or more of the following positions at 2θ: 5.2±0.2°, 7.4±0.2°, 14.5±0.2°, 16.1±0.2°, 18.7±0.2°, 21.8±0.2°, 11.7±0.2°, 10.0±0.2°, 26.0±0.2°, and 20.4±0.2°. Preferably, diffraction peaks are found at 4, 6, 8, or 10 of these positions. For example, the X-ray powder diffraction pattern of potassium salt crystal form B of compound 1 has diffraction peaks at the following positions at 2θ: At angles of 5.2±0.2°, 7.4±0.2°, 14.5±0.2°, 16.1±0.2°, 18.7±0.2°, 21.8±0.2°, 11.7±0.2°, and 10.0±0.2°, Alternatively, at 5.2±0.2°, 7.4±0.2°, 14.5±0.2°, 16.1±0.2°, 18.7±0.2°, 21.8±0.2°, 11.7±0.2°, 10.0±0.2°, 26.0±0.2°, and 20.4±0.2°; The powder X-ray diffraction pattern of sodium salt crystal form 1 of compound 1 has characteristic peaks at one or more of the following positions at 2θ: 8.9±0.2°, 10.0±0.2°, 20.1±0.2°, 9.6±0.2°, 16.8±0.2°, 15.3±0.2°, 31.0±0.2°, 26.7±0.2°, 11.4±0.2°, and 24.2±0.2°; preferably, diffraction peaks are found at any of 4, 6, 8, or 10 of these positions. For example, the X-ray powder diffraction pattern of sodium salt crystal form 1 of compound 1 has diffraction peaks at the following positions at 2θ: At angles of 8.9±0.2°, 10.0±0.2°, 20.1±0.2°, 9.6±0.2°, 16.8±0.2°, 15.3±0.2°, 31.0±0.2°, and 26.7±0.2°, Alternatively, at 8.9±0.2°, 10.0±0.2°, 20.1±0.2°, 9.6±0.2°, 16.8±0.2°, 15.3±0.2°, 31.0±0.2°, 26.7±0.2°, 11.4±0.2°, and 24.2±0.2°; The X-ray powder diffraction pattern of sodium salt crystal form 2 of compound 1 has diffraction peaks at one or more of the following positions at 2θ: 10.3±0.2°, 20.5±0.2°, 8.9±0.2°, 7.8±0.2°, 17.1±0.2°, 15.3±0.2°, 26.6±0.2°, 24.2±0.2°, 12.9±0.2°, and 11.9±0.2°; preferably, it includes diffraction peaks at any of 4, 6, 8, or 10 of these positions. For example, the X-ray powder diffraction pattern of sodium salt crystal form 2 of compound 1 has diffraction peaks at the following positions at 2θ: At angles of 10.3±0.2°, 20.5±0.2°, 8.9±0.2°, 7.8±0.2°, 17.1±0.2°, 15.3±0.2°, 26.6±0.2°, and 24.2±0.2°, Alternatively, at 10.3±0.2°, 20.5±0.2°, 8.9±0.2°, 7.8±0.2°, 17.1±0.2°, 15.3±0.2°, 26.6±0.2°, 24.2±0.2°, 12.9±0.2°, and 11.9±0.2°; The X-ray powder diffraction pattern of sodium salt crystal form 1 of compound 2 has diffraction peaks at one or more of the following 2θ values: 10.4±0.2°, 5.3±0.2°, 9.0±0.2°, 7.7±0.2°, 20.5±0.2°, 17.4±0.2°, 24.9±0.2°, 15.0±0.2°, 11.9±0.2°, 21.2±0.2°, 19.9±0.2°, 16.5±0.2°, 18.0±0.2°, 15.9±0.2°, and 26.8±0.2°; preferably, diffraction peaks are found at any of 4, 6, 8, or 10 of these values; for example, the X-ray powder diffraction pattern of sodium salt crystal form 1 of compound 2 has diffraction peaks at the following 2θ positions: At angles of 10.4±0.2°, 5.3±0.2°, 9.0±0.2°, 7.7±0.2°, 20.5±0.2°, 17.4±0.2°, 24.9±0.2°, and 15.0±0.2°, Alternatively, at 10.4±0.2°, 5.3±0.2°, 9.0±0.2°, 7.7±0.2°, 20.5±0.2°, 17.4±0.2°, 24.9±0.2°, 15.0±0.2°, and 11.9±0.2°, The X-ray powder diffraction pattern of potassium salt crystal form 1 of compound 2 has diffraction peaks at one or more of the following positions at 2θ: 9.8±0.2°, 19.5±0.2°, 26.7±0.2°, 13.0±0.2°, 15.6±0.2°, 21.2±0.2°, 24.3±0.2°, 8.6±0.2°, 22.7±0.2°, 20.7±0.2°, 16.3±0.2°, 24.6±0.2°, 29.4±0.2°, 30.3±0.2°, and 15.0±0.2°; preferably, it includes diffraction peaks at any of 4, 6, 8, or 10 positions; for example, the X-ray powder diffraction pattern of potassium salt crystal form 1 of compound 2 has diffraction peaks at the following positions at 2θ: At 9.8±0.2°, 19.5±0.2°, 26.7±0.2°, 13.0±0.2°, 15.6±0.2°, 21.2±0.2°, 24.3±0.2°, and 8.6±0.2°, Alternatively, at 9.8±0.2°, 19.5±0.2°, 26.7±0.2°, 13.0±0.2°, 15.6±0.2°, 21.2±0.2°, 24.3±0.2°, 8.6±0.2°, 22.7±0.2°, and 20.7±0.2°.

9. The acidic salt crystal form or the basic salt crystal form according to claim 6, characterized in that, The X-ray powder diffraction pattern of sulfate crystal form A of compound 1 is shown in Figure 1; The X-ray powder diffraction pattern of potassium salt crystal form A of compound 1 is shown in Figure 2; The X-ray powder diffraction pattern of potassium salt form B of compound 1 is shown in Figure 3; The X-ray powder diffraction pattern of sodium salt crystal form 1 of compound 1 is shown in Figure 4; The X-ray powder diffraction pattern of sodium salt crystal form 2 of compound 1 is shown in Figure 5; The X-ray powder diffraction pattern of sodium salt crystal form 1 of compound 2 is shown in Figure 6; The X-ray powder diffraction pattern of potassium salt crystal form 1 of compound 2 is shown in Figure 7.

10. The acid salt crystal form or basic salt crystal form according to any one of claims 1-9, characterized in that, The 2θ error between the positions of the top ten diffraction peaks with relative intensities in the X-ray powder diffraction pattern of the sulfate crystal form A of compound 1 and the corresponding diffraction peaks in Figure 1 is ±0.2° to ±0.5°, preferably ±0.2° to ±0.3°, and more preferably ±0.2°; the 2θ error between the positions of the top ten diffraction peaks with relative intensities in the X-ray powder diffraction pattern of the potassium salt crystal form A of compound 1 and the corresponding diffraction peaks in Figure 2 is ±0.2° to ±0.5°. Preferably ±0.2° to ±0.3°, more preferably ±0.2°; the 2θ error between the positions of the top ten diffraction peaks with the highest relative intensities in the X-ray powder diffraction pattern of potassium salt form B of compound 1 and the corresponding positions in Figure 3 is ±0.2° to ±0.5°, preferably ±0.2° to ±0.3°, more preferably ±0.2°; the positions of the top ten diffraction peaks with the highest relative intensities in the X-ray powder diffraction pattern of sodium salt form 1 of compound 1 and the corresponding positions in Figure 4 are... The 2θ error of the diffraction peaks is ±0.2° to ±0.5°, preferably ±0.2° to ±0.3°, and more preferably ±0.2°; the 2θ error between the positions of the top ten diffraction peaks with the highest relative intensities in the X-ray powder diffraction pattern of sodium salt crystal form 2 of compound 1 and the corresponding positions in Figure 5 is ±0.2° to ±0.5°, preferably ±0.2° to ±0.3°, and more preferably ±0.2°; the X-ray powder diffraction pattern of sodium salt crystal form 1 of compound 2. The 2θ error between the positions of the top ten diffraction peaks with relative peak intensities in the X-ray powder diffraction pattern of the potassium salt crystal form 1 of compound 2 and the corresponding diffraction peaks in Figure 6 is ±0.2° to ±0.5°, preferably ±0.2° to ±0.3°, and more preferably ±0.2°.

11. A method for preparing the acid salt or basic salt crystal form of the compound according to any one of claims 1-10, specifically comprising the following steps: 1) Weigh an appropriate amount of compound (I) and dissolve it in a good solvent; 2) Weigh an appropriate amount of the counter-ion base and dissolve it in an organic solvent; the amount of the counter-ion base is preferably 1.2 equivalents; 3) Combine the two solutions and stir to precipitate. 4) Dry to obtain the target product; in: The beneficial solvent is selected from methanol, ethyl acetate, acetone, tetrahydrofuran, isopropanol, or 2-butanone; preferably ethyl acetate or tetrahydrofuran. The organic solvent is selected from methanol, ethanol, ethyl acetate, dichloromethane, acetone, acetonitrile, tetrahydrofuran, 2-butanone, 3-pentanone, tert-butanol, or N,N-dimethylformamide; ethanol is preferred; the above-mentioned benign solvents and organic solutions must be miscible when used. The counterion base is an organic or inorganic base. The organic base is selected from sodium methoxide, potassium ethoxide, trimethylamine, diethylamine, triethylamine, triethanolamine, pyridine, piperidine, morpholine, diisopropylethylamine, diisopropylaminolithium, diethylaminolithium, bis(trimethylsilyl)aminolithium, potassium acetate, sodium acetate, isopropylcyclohexylaminolithium, or mixtures thereof. The inorganic base is selected from potassium phosphate, potassium phosphate trihydrate, potassium phosphate dihydrate, potassium phosphate monohydrate, sodium bicarbonate, potassium bicarbonate, sodium carbonate, cesium carbonate, potassium hydroxide, sodium hydroxide, potassium hydride, sodium hydride, lithium hydroxide, or mixtures thereof. Preferred bases are sodium methoxide or potassium hydroxide.

12. A method for preparing the crystal form of the compound according to any one of claims 1-10, wherein the crystal form is a sodium salt crystal form, specifically comprising the following steps: 1) Weigh an appropriate amount of the sodium salt of the compound and suspend it in a poor solvent. The preferred suspension density is 50-200 mg / mL. 2) The suspension obtained above is stirred at a certain temperature for a certain time, preferably 0-50℃, and the time is preferably 1-10 days; 3) The above suspension was rapidly centrifuged to remove the supernatant. The remaining solid was placed in a vacuum drying oven at 40°C and dried to constant weight to obtain the target product. in: The undesirable solvent is selected from methyl tert-butyl ether, isopropyl ether, diethyl ether, n-hexane, n-heptane, cyclohexane, n-pentane, water, and a mixture of water and the aforementioned benign solvents; preferably methyl tert-butyl ether, isopropyl ether, and n-heptane.

13. A method for preparing the acid salt or basic salt crystal form of the compound according to any one of claims 1-10, specifically comprising the following steps: 1) Weigh an appropriate amount of compound (I) and dissolve it in a good solvent; 2) Weigh an appropriate amount of the counterionic acid and dissolve it in an organic solvent; 3) Combine the two solutions and stir to precipitate. 4) Dry to obtain the target product; in: The beneficial solvent is selected from methanol, ethyl acetate, acetone, tetrahydrofuran, isopropanol, or 2-butanone; tetrahydrofuran is preferred. The organic solvent is selected from methanol, ethanol, ethyl acetate, dichloromethane, acetone, acetonitrile, tetrahydrofuran, 2-butanone, 3-pentanone, tert-butanol, or N,N-dimethylformamide; ethanol is preferred; the above-mentioned benign solvents and organic solutions must be miscible when used. The aforementioned counterionic acid is selected from inorganic or organic acids, wherein the inorganic acid is selected from hydrochloric acid, sulfuric acid, nitric acid, hydrobromic acid, hydrofluoric acid, hydroiodic acid, or phosphoric acid; and the organic acid is selected from 2,5-dihydroxybenzoic acid, 1-hydroxy-2-naphtholic acid, acetic acid, dichloroacetic acid, trichloroacetic acid, acetoxyxamic acid, adipic acid, benzenesulfonic acid, 4-chlorobenzenesulfonic acid, benzoic acid, 4-acetaminobenzoic acid, 4-aminobenzoic acid, decanoic acid, hexanoic acid, caprylic acid, cinnamic acid, citric acid, cyclohexanesulfonic acid, camphorsulfonic acid, aspartic acid, camphoric acid, gluconic acid, glucuronic acid, glutamic acid, isoascorbic acid, lactic acid, malic acid, mandelic acid, pyroglutamic acid, tartaric acid, etc. Dodecyl sulfuric acid, dibenzoyl tartaric acid, ethane-1,2-disulfonic acid, ethanesulfonic acid, formic acid, fumaric acid, galactobionic acid, gentian acid, glutaric acid, 2-ketoglutaric acid, glycolic acid, hippuric acid, hydroxyethyl sulfonic acid, lactobionic acid, ascorbic acid, aspartic acid, lauric acid, camphoric acid, maleic acid, malonic acid, methanesulfonic acid, 1,5-naphthalenedisulfonic acid, naphthalene-2-sulfonic acid, nicotinic acid, oleic acid, orotic acid, oxalic acid, palmitic acid, dihydroxynaphthalic acid, propionic acid, salicylic acid, 4-aminosalicylic acid, sebacic acid, stearic acid, succinic acid, thiocyanate, undecanoic acid, trifluoroacetic acid, benzenesulfonic acid, p-methylbenzenesulfonic acid, or L-malic acid; the counterionic acid is preferably sulfuric acid.

14. A pharmaceutical composition comprising a therapeutically effective amount of the acidic or basic salt form of the compound of any one of claims 1-10, and one or more pharmaceutically acceptable carriers, diluents, or excipients.

15. A method for preparing a pharmaceutical composition comprising the compound of claim 1 or a pharmaceutically acceptable salt thereof, comprising the step of mixing an acidic or basic salt form of the compound of any one of claims 1-10 with one or more pharmaceutically acceptable carriers, diluents or excipients.

16. Use of the acid salt or basic salt crystal form of the compound of any one of claims 1-10, or the pharmaceutical composition of claim 14, in the preparation of a medicament for treating cancer.

17. Use of the acidic or basic salt form of the compound of any one of claims 1-10, or the pharmaceutical composition of claim 14, in the preparation of a medicament for treating a condition or disease that can be treated by WRN inhibition.

18. The use according to claim 16 or 17, wherein the cancer or the condition or disease is selected from the group consisting of microsatellite instability-high (MSI-H) or mismatch repair deficiency (dMMR), preferably, microsatellite instability-high (MSI-H) or mismatch repair deficiency (dMMR) selected from colorectal cancer, gastric cancer, prostate cancer, endometrial cancer, adrenocortical cancer, uterine cancer, cervical cancer, esophageal cancer, breast cancer, kidney cancer, and ovarian cancer.

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