A selective inhibitor for brca mutations and uses thereof

Abstract

The application discloses a selective inhibitor for BRCA mutation and application thereof. Specifically, the application relates to a quinazolinone compound shown in a general formula (I) or a pharmaceutically acceptable salt or a solvate thereof, wherein the compound has better selectivity for BRCA mutation, has a good inhibition rate on the proliferation of BRCA mutant cells at a low concentration, and is superior to existing olaparib and AZD5305 and the like. Through a nude mouse tumorigenesis experiment, it is proved that the drug obviously shows an inhibitory effect on tumor growth at a low drug concentration, and has no obvious toxic side effects.

Description

Technical Field

[0001] This invention belongs to the field of pharmaceutical technology, specifically relating to a selective inhibitor of BRCA mutations and its uses. Background Technology

[0002] Poly(ADP-ribose) polymerases (PARPs) are ribozymes found in eukaryotic cells, primarily involved in DNA repair and maintaining genome stability. Among the 18 members of the PARP family, PARP-1 has the highest intracellular content and has been the most extensively studied, mainly through its NAD+ expression. + PARP-1 acts as a substrate catalyzer for the transfer of ADP-ribose units to nuclear receptor proteins, forming poly-ADP-ribose polymers that guide repair enzymes to repair DNA gaps, playing a crucial role in the base excision repair pathway. Given the current scarcity of high-quality drug targets, PARP-1 provides one of the few clinically validated anticancer targets. PARP-1 inhibitors exert their targeting effect by cleaving bases in homologous recombination repair-deficient tumor cells, thus excising the repair pathway. Therefore, they can be used alone in such tumors, such as BRCA1 / 2-deficient breast and ovarian cancer.

[0003] BRCA1 and BRCA2 genes are important tumor suppressor genes in the human body, responsible for repairing DNA damage that occurs in cells. BRCA mutations refer to mutations or variations in breast cancer-related genes (BRCA1 and BRCA2 genes). BRCA mutations may impair DNA repair mechanisms, thereby increasing the risk of cells suffering further DNA damage. This DNA damage accumulates gradually in cells, potentially leading to abnormal cell growth and tumor formation.

[0004] In December 2014, Olaparib was first approved for the treatment of platinum-sensitive recurrent ovarian cancer with BRCA1 / 2 deficiency. Clinical studies are also underway for its use alone or in combination with other indications with homologous recombination repair deficiency, such as breast cancer, pancreatic cancer, and prostate cancer.

[0005] To date, following the FDA approval of AstraZeneca's first PARP inhibitor, olaparib, five more PARP inhibitors have been approved: rucaparib (Clovis Oncology, 2016), niraparib (Tesaro / Zai Lab, 2017 / 2019), tapolazoparib (Pfizer, 2018), fluzoparib (Hengrui Medicine / Hausen, 2020), and pamiparib (BeiGene, 2021). These first-generation PARP inhibitors primarily utilize high-throughput fragment screening or crystal structure-based methods for structural design, focusing on enhancing the inhibitory activity of compounds against PARP1 / 2 enzymes, improving physicochemical properties, and expanding their combination with other chemotherapy drugs and methods to better treat breast and ovarian cancers with BRCA1 / 2 deficiencies.

[0006] However, while these drugs have shown some therapeutic effect, their use in treating BRCA-mutated diseases presents the challenge of requiring high concentrations to achieve optimal results. Higher doses, however, undoubtedly increase the risk of side effects and treatment costs, thus limiting their widespread application.

[0007] Therefore, there is a need to provide an improved technical solution that addresses the shortcomings of the existing technology. Summary of the Invention

[0008] The purpose of this invention is to provide a selective inhibitor against BRCA mutations, which has better selectivity for diseases caused by BRCA mutations, in order to solve the problems of high drug concentration and high risk of side effects of existing PARP inhibitors.

[0009] To achieve the above objectives, the present invention provides the following technical solution:

[0010] A selective inhibitor against BRCA mutations, comprising a compound of formula (I) or a pharmaceutically acceptable salt thereof or a solvation thereof.

[0011]

[0012] Wherein, R1 is selected from

[0013]

[0014]

[0015] R2 is selected from phenyl, pyridyl, indolyl, naphthyl, phenyl with one or more substituents, pyridyl with one or more substituents, indolyl with one or more substituents, or naphthyl with one or more substituents;

[0016] R3 is selected from H, alkyl, halogen, cyano, hydroxyl, alkoxy, carboxyl, amide, carboxylic acid ester, amino, nitro, trifluoromethyl, trifluoromethoxy, haloalkyl, hydroxyalkyl, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, hydroxyalkyl ether, aryl with one or more substituents, or heteroaryl with one or more substituents.

[0017] X is selected from O or S;

[0018] A, B, C, and D are each independently selected from H or N.

[0019] Preferably, R2 is selected from phenyl with one or more substituents, pyridyl with one or more substituents, indole with one or more substituents, and naphthyl with one or more substituents, wherein the substituents on the phenyl, pyridyl, indole, and naphthyl are each independently selected from alkyl, halogen, cyano, hydroxy, alkoxy, carboxyl, amide, carboxylate, amino, nitro, trifluoromethyl, oxotrifluoromethyl, haloalkyl, hydroxyalkyl, cycloalkyl, heterocycloalkyl, aryl, and heteroaryl.

[0020] Preferably, R3 is selected from alkyl groups, halogens, and methoxy groups.

[0021] Preferably, the compound represented by formula (I) or its pharmaceutically acceptable salt or solvate is one of the following compounds:

[0022]

[0023]

[0024] Preferably, a compound of formula (I) or a pharmaceutically acceptable salt thereof or a solvate thereof,

[0025]

[0026] Wherein, R2 is selected from

[0027]

[0028] Preferably, the compound is N-(5-chloro-2-methylphenyl)-3-((4-oxo-3,4-dihydroquinazolin-2-yl)methyl)-3,6-diazabicyclo[3.1.1]heptane-6-carboxamide.

[0029] Preferably, the compound is N-(5-chloro-2-methylphenyl)-4-((4-oxo-3,4-dihydroquinazolin-2-yl)methyl)-1,4-diazacycloheptane-1-carboxamide.

[0030] The present invention also proposes a pharmaceutical composition comprising any of the above-described compounds or their pharmaceutically acceptable salts or solvates, and at least one pharmaceutically acceptable excipient.

[0031] The present invention also proposes the use of any of the above-mentioned compounds or their pharmaceutically acceptable salts or solvates in the preparation of medicaments for treating cancer.

[0032] Preferably, the cancer is selected from breast cancer, ovarian cancer, lung cancer, pancreatic cancer, gastric cancer, breast cancer, brain tumor, melanoma, renal cell carcinoma, colorectal cancer, head and neck cancer, bladder cancer, prostate cancer, hepatocellular carcinoma, or bile duct cancer.

[0033] Beneficial effects:

[0034] (1) The selective inhibitor provided by the present invention has good selectivity for BRCA mutant and non-mutated cells, especially for BRCA mutant cells, with a good inhibition rate at low concentrations, which is superior to olaparib and AZD5305, and helps to reduce drug concentration and the incidence of side effects.

[0035] (2) The selective inhibitors provided by the present invention can inhibit the cell proliferation of HCC1937BRCA1(- / -) cell line by more than 66%, inhibit the cell proliferation of HCT-15BRCA2(- / -) cell line by more than 70%, and inhibit the cell proliferation of Capan-1BRCA2(- / -) cell line by more than 50%, which are significantly better than the cell proliferation inhibition effects of existing compounds. Attached Figure Description

[0036] The accompanying drawings, which form part of this application, are used to provide a further understanding of the invention. The illustrative embodiments of the invention and their descriptions are used to explain the invention and do not constitute an undue limitation of the invention. Wherein:

[0037] Figure 1 The change in body weight of mice after the start of drug administration in a mouse tumorigenesis experiment.

[0038] Figure 2 The relative volume change of tumors after drug administration was initiated in a mouse tumorigenesis experiment.

[0039] Figure 3 The changes in tumor weight in mice in the treatment group and the control group.

[0040] Figure 4 Photos of tumors in mice in the treatment and control groups. Detailed Implementation

[0041] The technical solutions in the embodiments of the present invention will be clearly and completely described below. Obviously, the described embodiments are only a part of the embodiments of the present invention, and not all of them. All other embodiments obtained by those skilled in the art based on the embodiments of the present invention are within the scope of protection of the present invention.

[0042] This invention addresses the problems of high drug concentrations and significant side effects associated with existing PARP inhibitors by providing a selective inhibitor against BRCA mutations, which is a compound having the following formula (Ⅰ) or a pharmaceutically acceptable salt or solvate thereof.

[0043]

[0044] Wherein, R1 is selected from

[0045]

[0046]

[0047] R2 is selected from phenyl, pyridyl, indolyl, naphthyl, phenyl with one or more substituents, pyridyl with one or more substituents, indolyl with one or more substituents, or naphthyl with one or more substituents;

[0048] R3 is selected from H, alkyl, halogen, cyano, hydroxyl, alkoxy, carboxyl, amide, carboxylic acid ester, amino, nitro, trifluoromethyl, trifluoromethoxy, haloalkyl, hydroxyalkyl, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, hydroxyalkyl ether, aryl with one or more substituents, or heteroaryl with one or more substituents.

[0049] X is selected from O or S;

[0050] A, B, C, and D are each independently selected from H or N.

[0051] Specifically, the present invention relates to the following compounds or pharmaceutically acceptable salts or solvates thereof:

[0052] N-(2-methyl-5-chlorophenyl)-4-((4-oxo-3,4-dihydroquinazolin-2-yl)methyl)piperazine-1-carboxamide;

[0053] N-(2,3-difluorophenyl)-4-((4-oxo-3,4-dihydroquinazolin-2-yl)methyl)piperazine-1-carboxamide;

[0054] N-(3,4-dimethylphenyl)-4-((4-oxo-3,4-dihydroquinazolin-2-yl)methyl)piperazine-1-carboxamide;

[0055] N-(3,5-dimethylphenyl)-4-((4-oxo-3,4-dihydroquinazolin-2-yl)methyl)piperazine-1-carboxamide;

[0056] N-(2-methyl-5-chlorophenyl)-3-((4-oxo-3,4-dihydroquinazolin-2-yl)methyl)-3,6-diazabicyclo[3.1.1]heptane-6-carboxamide;

[0057] N-(5-chloro-2-methylphenyl)-2-methyl-4-((4-oxo-3,4-dihydroquinazolin-2-yl)methyl)piperazine-1-carboxamide;

[0058] (1S,4S)-N-(5-chloro-2-methylphenyl)-5-((4-oxo-3,4-dihydroquinazolin-2-yl)methyl)-2,5-diazabicyclo[2.2.1]heptane-2-carboxamide;

[0059] N-(5-chloro-2-methylphenyl)-4-((4-oxo-3,4-dihydroquinazolin-2-yl)methyl)-1,4-diazacycloheptane-1-carboxamide;

[0060] N-(5-chloro-2-methylphenyl)-4-((4-oxo-3,4-dihydroquinazolin-2-yl)methyl)piperazine-1-thioamide;

[0061] N-(5-chloro-2-methylphenyl)-3-((7-fluoro-4-oxo-3,4-dihydroquinazolin-2-yl)methyl)-3,6-diazabicyclo[3.1.1]heptane-6-carboxamide;

[0062] N-(5-chloro-2-methylphenyl)-3-(((4-oxo-3,4-dihydroquinazolin-2-yl)methyl)amino)azacyclobut-1-carboxamide;

[0063] (1R,5S)-N-(5-chloro-2-methylphenyl)-8-((4-oxo-3,4-dihydroquinazolin-2-yl)methyl)-3,8-diazabicyclo[3.2.1]octane-3-carboxamide;

[0064] N-(5-chloro-2-methylphenyl)-3-((4-oxo-3,4-dihydroquinazolin-2-yl)methyl)-3,8-diazabicyclo[3.2.1]octane-8-carboxamide;

[0065] N-(5-chloro-2-methylphenyl)-3-((4-oxo-3,4-dihydroquinazolin-2-yl)methyl)tetrahydropyrimidine-1(2H)-formamide;

[0066] N-(3-chloro-4-fluorophenyl)-4-((4-oxo-3,4-dihydroquinazolin-2-yl)methyl)piperazine-1-carboxamide;

[0067] N-(2,4-difluorophenyl)-4-((4-oxo-3,4-dihydroquinazolin-2-yl)methyl)piperazine-1-carboxamide;

[0068] N-(5-fluoro-2-methylphenyl)-4-((4-oxo-3,4-dihydroquinazolin-2-yl)methyl)piperazine-1-carboxamide;

[0069] N-(2,5-difluorophenyl)-4-((4-oxo-3,4-dihydroquinazolin-2-yl)methyl)piperazine-1-carboxamide;

[0070] N-(3-fluoro-2-methylphenyl)-4-((4-oxo-3,4-dihydroquinazolin-2-yl)methyl)piperazine-1-carboxamide;

[0071] N-(3,5-difluorophenyl)-4-((4-oxo-3,4-dihydroquinazolin-2-yl)methyl)piperazine-1-carboxamide;

[0072] N-(2-fluoro-5-methylphenyl)-4-((4-oxo-3,4-dihydroquinazolin-2-yl)methyl)piperazine-1-carboxamide;

[0073] N-(4-fluoro-2-methylphenyl)-4-((4-oxo-3,4-dihydroquinazolin-2-yl)methyl)piperazine-1-carboxamide;

[0074] N-(3,5-dichlorophenyl)-4-((4-oxo-3,4-dihydroquinazolin-2-yl)methyl)piperazine-1-carboxamide;

[0075] N-(2-chloro-6-fluorophenyl)-4-((4-oxo-3,4-dihydroquinazolin-2-yl)methyl)piperazine-1-carboxamide;

[0076] N-(2-chloro-5-(trifluoromethyl)phenyl)-4-((4-oxo-3,4-dihydroquinazolin-2-yl)methyl)piperazine-1-carboxamide;

[0077] N-(2-chloro-5-methylphenyl)-4-((4-oxo-3,4-dihydroquinazolin-2-yl)methyl)piperazine-1-carboxamide;

[0078] N-(3-bromo-2-methylphenyl)-4-((4-oxo-3,4-dihydroquinazolin-2-yl)methyl)piperazine-1-carboxamide;

[0079] N-(2-methoxy-5-(trifluoromethyl)phenyl)-4-((4-oxo-3,4-dihydroquinazolin-2-yl)methyl)piperazine-1-carboxamide;

[0080] N-(2-fluoro-4-(trifluoromethyl)phenyl)-4-((4-oxo-3,4-dihydroquinazolin-2-yl)methyl)piperazine-1-carboxamide;

[0081] N-(3,5-bis(trifluoromethyl)phenyl)-4-((4-oxo-3,4-dihydroquinazolin-2-yl)methyl)piperazine-1-carboxamide;

[0082] N-(2,6-diisopropylphenyl)-4-((4-oxo-3,4-dihydroquinazolin-2-yl)methyl)piperazine-1-carboxamide;

[0083] N-(2,6-dimethylphenyl)-4-((4-oxo-3,4-dihydroquinazolin-2-yl)methyl)piperazine-1-carboxamide;

[0084] N-(2-fluoro-4-methylphenyl)-4-((4-oxo-3,4-dihydroquinazolin-2-yl)methyl)piperazine-1-carboxamide;

[0085] N-(2-fluoro-3-methylphenyl)-4-((4-oxo-3,4-dihydroquinazolin-2-yl)methyl)piperazine-1-carboxamide;

[0086] N-(2-Fluorophenyl)-4-((4-oxo-3,4-dihydroquinazolin-2-yl)methyl)piperazine-1-carboxamide;

[0087] N-(3-Fluorophenyl)-4-((4-oxo-3,4-dihydroquinazolin-2-yl)methyl)piperazine-1-carboxamide;

[0088] N-(4-fluorophenyl)-4-((4-oxo-3,4-dihydroquinazolin-2-yl)methyl)piperazine-1-carboxamide;

[0089] N-(2-chlorophenyl)-4-((4-oxo-3,4-dihydroquinazolin-2-yl)methyl)piperazine-1-carboxamide;

[0090] N-(3-chlorophenyl)-4-((4-oxo-3,4-dihydroquinazolin-2-yl)methyl)piperazine-1-carboxamide;

[0091] N-(4-chlorophenyl)-4-((4-oxo-3,4-dihydroquinazolin-2-yl)methyl)piperazine-1-carboxamide;

[0092] N-(2-bromophenyl)-4-((4-oxo-3,4-dihydroquinazolin-2-yl)methyl)piperazine-1-carboxamide;

[0093] N-(3-bromophenyl)-4-((4-oxo-3,4-dihydroquinazolin-2-yl)methyl)piperazine-1-carboxamide;

[0094] N-(4-bromophenyl)-4-((4-oxo-3,4-dihydroquinazolin-2-yl)methyl)piperazine-1-carboxamide;

[0095] N-(2-aminophenyl)-4-((4-oxo-3,4-dihydroquinazolin-2-yl)methyl)piperazine-1-carboxamide;

[0096] N-(3-aminophenyl)-4-((4-oxo-3,4-dihydroquinazolin-2-yl)methyl)piperazine-1-carboxamide;

[0097] N-(4-aminophenyl)-4-((4-oxo-3,4-dihydroquinazolin-2-yl)methyl)piperazine-1-carboxamide;

[0098] N-(2-nitrophenyl)-4-((4-oxo-3,4-dihydroquinazolin-2-yl)methyl)piperazine-1-carboxamide;

[0099] N-(3-nitrophenyl)-4-((4-oxo-3,4-dihydroquinazolin-2-yl)methyl)piperazine-1-carboxamide;

[0100] N-(4-nitrophenyl)-4-((4-oxo-3,4-dihydroquinazolin-2-yl)methyl)piperazine-1-carboxamide;

[0101] 4-((4-oxo-3,4-dihydroquinazolin-2-yl)methyl)-N-(2-trifluoromethylphenyl)piperazine-1-carboxamide;

[0102] 4-((4-oxo-3,4-dihydroquinazolin-2-yl)methyl)-N-(3-trifluoromethylphenyl)piperazine-1-carboxamide;

[0103] 4-((4-oxo-3,4-dihydroquinazolin-2-yl)methyl)-N-(4-trifluoromethylphenyl)piperazine-1-carboxamide;

[0104] N-(2-methoxyphenyl)-4-((4-oxo-3,4-dihydroquinazolin-2-yl)methyl)piperazine-1-carboxamide;

[0105] N-(3-methoxyphenyl)-4-((4-oxo-3,4-dihydroquinazolin-2-yl)methyl)piperazine-1-carboxamide;

[0106] N-(4-methoxyphenyl)-4-((4-oxo-3,4-dihydroquinazolin-2-yl)methyl)piperazine-1-carboxamide;

[0107] N-(3-cyanophenyl)-4-((4-oxo-3,4-dihydroquinazolin-2-yl)methyl)piperazine-1-carboxamide;

[0108] N-(4-cyanophenyl)-4-((4-oxo-3,4-dihydroquinazolin-2-yl)methyl)piperazine-1-carboxamide;

[0109] N-(2-tert-butylphenyl)-4-((4-oxo-3,4-dihydroquinazolin-2-yl)methyl)piperazine-1-carboxamide;

[0110] N-(4-acetylphenyl)-4-((4-oxo-3,4-dihydroquinazolin-2-yl)methyl)piperazine-1-carboxamide;

[0111] N-(4-Ethylphenyl)-4-((4-oxo-3,4-dihydroquinazolin-2-yl)methyl)piperazine-1-carboxamide;

[0112] N-(5-chloro-2-methylphenyl)-4-((4-oxo-3,4-dihydroquinazolin-2-yl)methyl)piperazine-1-carboxamide.

[0113] The following detailed description of a PARP1 selective inhibitor of the present invention, its preparation method, and its uses is illustrated by specific embodiments.

[0114] Example 1

[0115] Preparation of N-(5-chloro-2-methylphenyl)-4-((4-oxo-3,4-dihydroquinazolin-2-yl)methyl)piperazine-1-carboxamide:

[0116]

[0117] Preparation route:

[0118]

[0119] Step 1: Synthesis of 2-(chloromethyl)quinazoline-4-(3H)-quinazolineone:

[0120]

[0121] At room temperature, methyl 2-aminobenzoate (1 eq) and chloroacetonitrile (3 eq) were placed in a reaction tube, followed by the addition of 4N hydrogen chloride-dioxane solution. The mixture was heated to 80 °C and stirred for 24 h. After 24 hours of reaction, the reaction solution was cooled to room temperature, the resulting solid was collected and dissolved in water, neutralized with sodium hydroxide to pH = 7, the solid was collected by filtration, washed with water and dried, and finally 6.05 g of the target product was obtained as a white solid with a yield of 65.3% and a purity of 95.2%. MS m / z (ESI): 195.2 [M+1].

[0122] Step 2: Synthesis of tert-butyl 4-((4-oxo-3,4-dihydroquinazolin-2-yl)methyl)piperazine-1-carboxylic acid:

[0123]

[0124] At room temperature, 2-chloromethyl-4-(3H)-quinazolinone (1 eq), N-Boc-piperazine (2 eq), and potassium carbonate (3 eq) were placed in a reaction tube, followed by the addition of dichloromethane. The mixture was stirred at room temperature for 12 h until the reaction was complete. The reaction was then quenched with water, extracted with ethyl acetate, and the organic phases were combined. The mixture was then washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and column chromatographically purified to give 5.83 g of the target product as a white solid with a yield of 53.5% and a purity of 96.1%. MS m / z (ESI): 345.4 [M+1].

[0125]

[0126] Step 3: Synthesis of 2-(piperazine-1-methyl)-4-(3H)-quinazolinone:

[0127] At room temperature, 4-((4-oxo-3,4-dihydroquinazolin-2-yl)methyl)piperazine-1-carboxylic acid tert-butyl ester (1 eq) was placed in a reaction tube, followed by the addition of dichloromethane and then a 4N hydrogen chloride-dioxane solution. The mixture was stirred at room temperature until the reaction was complete. Sodium hydroxide was added dropwise until the pH reached 8-10. The solid was filtered and dried to obtain 2.3 g of the target product as a white solid with a yield of 63.2% and a purity of 95.7%. MS m / z (ESI): 245.5 [M+1].

[0128] Step 4: Synthesis of N-(5-chloro-2-methylphenyl)-4-((4-oxo-3,4-dihydroquinazolin-2-yl)methyl)piperazine-1-carboxamide (Compound 1):

[0129]

[0130] At room temperature, 2-(piperazin-1-methyl)-4-(3H)-quinazolinone (1 eq), 5-chloro-2-methylphenyl isocyanate (2 eq), and triethylamine (3 eq) were placed in a reaction tube, followed by the addition of tetrahydrofuran. The mixture was stirred at room temperature for 12 h until the reaction was complete. The mixture was then extracted with dichloromethane and water. The organic phases were combined, and the final product was obtained by column chromatography as a white solid, yielding 0.16 g of the target compound. The yield was 45.6%, and the purity was 98.7%. MS m / z (ESI): 412.9 [M+1]. 1 H NMR (400MHz, CDCl3) δ9.90 (s, 1H), 8.27 (d, J = 8.0Hz, 1H), 7.84 (s, 1H), 7.75 (t, J =7.6Hz,1H),7.65(d,J=8.2Hz,1H),7.47(t,J=7.5Hz,1H),7.03(dd,J=25.7,8.1H z,2H),6.16(s,1H),4.19(d,J=5.2Hz,2H),3.80(s,2H),3.39(d,J=10.8Hz,2H),2 .99(d,J=10.8Hz,2H),2.63(d,J=6.9Hz,1H),2.18(s,3H),1.90(d,J=8.1Hz,1H).

[0131] Example 2

[0132] Preparation of N-(2,3-difluorophenyl)-4-((4-oxo-3,4-dihydroquinazolin-2-yl)methyl)piperazine-1-carboxamide:

[0133]

[0134] Preparation route:

[0135] Step 1: Synthesis of 2-(chloromethyl)quinazolin-4-(3H)-one, the specific process is the same as step 1 in Example 1.

[0136]

[0137] Step 2: Synthesis of 4-((4-oxo-3,4-dihydroquinazolin-2-yl)methyl)piperazine-1-carboxylic acid tert-butyl ester, the specific process is the same as step 2 of Example 1.

[0138] Step 3: Synthesis of 2-(piperazine-1-methyl)quinazolin-4-(3H)-one, the specific process is the same as step 3 of Example 1.

[0139] Step 4: Synthesis of N-(2,3-difluorophenyl)-4-((4-oxo-3,4-dihydroquinazolin-2-yl)methyl)piperazine-1-carboxamide:

[0140]

[0141] At room temperature, 2-(piperazine-1-methyl)-4-(3H)-quinazolinone (1 eq), 2,3-difluorophenyl isocyanate (2 eq), and triethylamine (3 eq) were placed in a reaction tube, followed by the addition of tetrahydrofuran. The mixture was stirred at room temperature for 12 h until the reaction was complete. The mixture was then extracted with dichloromethane and water. The organic phases were combined, and the final product was obtained by column chromatography as a white solid, yielding 0.17 g of the target compound. The yield was 51.5%, and the purity was 97.9%. MS m / z (ESI): 400.3 [M+1]. 1 H NMR (400MHz, CDCl3) δ10.07(s,1H),8.28(d,J=7.9Hz,1H),7.91–7.72(m,2H),7.68(d,J=8.1Hz,1H),7.50 (t,J=7.4Hz,1H),7.09–6.75(m,2H),6.58(d,J=54.3Hz,1H),3.63(d,J=9.9Hz,6H),2.68(d,J=4.3Hz,4H).

[0142] Example 3

[0143] Preparation of N-(3,4-dimethylphenyl)-4-((4-oxo-3,4-dihydroquinazolin-2-yl)methyl)piperazine-1-carboxamide:

[0144]

[0145] Preparation route:

[0146]

[0147] Step 1: Synthesis of 2-(chloromethyl)quinazolin-4(3H)-one, the specific process is the same as step 1 in Example 1.

[0148] Step 2: Synthesis of 4-((4-oxo-3,4-dihydroquinazolin-2-yl)methyl)piperazine-1-carboxylic acid tert-butyl ester, the specific process is the same as step 2 of Example 1.

[0149] Step 3: Synthesis of 2-(piperazine-1-methyl)quinazolin-4(3H)-one, the specific process is the same as step 3 of Example 1.

[0150] Step 4: Synthesis of N-(3,4-dimethylphenyl)-4-((4-oxo-3,4-dihydroquinazolin-2-yl)methyl)piperazine-1-carboxamide:

[0151]

[0152] At room temperature, 2-(piperazine-1-methyl)-4-(3H)-quinazolinone (1 eq), 3,4-dimethylphenyl isocyanate (2 eq), and triethylamine (3 eq) were placed in a reaction tube, followed by the addition of tetrahydrofuran. The mixture was stirred at room temperature for 12 h until the reaction was complete. The mixture was then extracted with dichloromethane and water. The organic phases were combined, and the final product was obtained by column chromatography as a white solid, yielding 0.16 g of the target compound. The yield was 44.6%, and the purity was 98.2%. MS m / z (ESI): 392.6 [M+1]. 1 H NMR (400MHz, CDCl3) δ9.93(s,1H),8.29(d,J=7.9Hz,1H),7.78(t,J=7.5Hz,1H),7.67(d,J=8.1Hz,1H),7.50(t,J =7.5Hz,1H),7.16(s,1H),7.04(s,2H),6.27(s,1H),3.61(d,J=23.6Hz,6H),2.66(s,4H),2.22(d,J=10.0Hz,6H).

[0153] Example 4

[0154] Preparation of N-(3,5-dimethylphenyl)-4-((4-oxo-3,4-dihydroquinazolin-2-yl)methyl)piperazine-1-carboxamide:

[0155]

[0156] Preparation route:

[0157]

[0158] Step 1: Synthesis of 2-(chloromethyl)quinazolin-4-(3H)-quinazolinone, the specific process is the same as step 1 in Example 1.

[0159] Step 2: Synthesis of 4-((4-oxo-3,4-dihydroquinazolin-2-yl)methyl)piperazine-1-carboxylic acid tert-butyl ester, the specific process is the same as step 2 of Example 1.

[0160] Step 3: Synthesis of 2-(piperazine-1-methyl)quinazolin-4(3H)-one, the specific process is the same as step 3 of Example 1.

[0161] Step 4: Synthesis of N-(3,5-dimethylphenyl)-4-((4-oxo-3,4-dihydroquinazolin-2-yl)methyl)piperazine-1-carboxamide:

[0162]

[0163] At room temperature, 2-(piperazin-1-methyl)-4-(3H)-quinazolinone (1 eq), 3,5-dimethylphenyl isocyanate (2 eq), and triethylamine (3 eq) were placed in a reaction tube, followed by the addition of tetrahydrofuran. The mixture was stirred at room temperature for 12 h until the reaction was complete. The mixture was then extracted with dichloromethane and water. The organic phases were combined, and the final product was obtained by column chromatography as a white solid, yielding 0.15 g of the target compound. The yield was 39.8%, and the purity was 97.7%. MS m / z (ESI): 392.6 [M+1]. 1 H NMR (400MHz, CDCl3) δ10.02(s,1H),8.28(d,J=7.9Hz,1H),7.65(ddd,J=74.4,15.0,7.5Hz,3 H), 6.98 (s, 2H), 6.69 (s, 1H), 6.40 (s, 1H), 3.60 (d, J = 20.2Hz, 6H), 2.63 (s, 4H), 2.27 (s, 6H).

[0164] Example 5

[0165] Preparation of N-(5-chloro-2-methylphenyl)-3-((4-oxo-3,4-dihydroquinazolin-2-yl)methyl)-3,6-diazabicyclo[3.1.1]heptane-6-carboxamide:

[0166]

[0167] Preparation route:

[0168]

[0169] Step 1: Synthesis of 2-(chloromethyl)quinazolin-4-(3H)-quinazolinone, the specific process is the same as step 1 in Example 1.

[0170] Step 2: Synthesis of tert-butyl 3-((4-oxo-3,4-dihydroquinazolin-2-yl)methyl)-3,6-diazabicyclo[3.1.1]heptane-6-carboxylic acid:

[0171]

[0172] At room temperature, 2-chloromethyl-4-(3H)-quinazolinone (1 eq), 6-N-Boc-3,6-diazabicyclo[3.1.1]heptane (2 eq), and potassium carbonate (3 eq) were placed in a reaction tube. Then, dichloromethane was added, and the mixture was stirred at room temperature for 12 h. After the reaction was complete, water was added to quench the reaction, and the mixture was extracted with ethyl acetate. The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and column chromatography was performed to obtain 6.01 g of the target product as a white solid with a yield of 55.6% and a purity of 96.7%. MS m / z (ESI): 357.6 [M+1].

[0173] Step 3: Synthesis of 2-((3,6-diazabicyclo[3.1.1]heptane-3-yl)methyl)-4(3H)-quinazolinone:

[0174]

[0175] At room temperature, 3-((4-oxo-3,4-dihydroquinazolin-2-yl)methyl)-3,6-diazabicyclo[3.1.1]heptane-6-carboxylic acid tert-butyl ester (1 eq) was placed in a reaction tube, followed by the addition of dichloromethane and then a 4N hydrogen chloride-dioxane solution. The mixture was stirred at room temperature until the reaction was complete. Sodium hydroxide was added dropwise until the pH reached 8-10. The solid was filtered and dried to obtain 2.4 g of the target product as a white solid with a yield of 64.7% and a purity of 97.1%. MS m / z (ESI): 257.4 [M+1].

[0176] Step 4: Synthesis of N-(5-chloro-2-methylphenyl)-3-((4-oxo-3,4-dihydroquinazolin-2-yl)methyl)-3,6-diazabicyclo[3.1.1]heptane-6-carboxamide:

[0177]

[0178] At room temperature, 2-((3,6-diazabicyclo[3.1.1]heptane-3-yl)methyl)-4(3H)-quinazolinone (1 eq), 5-chloro-2-methylphenyl isocyanate (2 eq), and triethylamine (3 eq) were placed in a reaction tube, followed by the addition of tetrahydrofuran. The mixture was stirred at room temperature for 12 h until the reaction was complete. The mixture was then extracted with dichloromethane and water. The organic phases were combined, and the final product was obtained by column chromatography as a white solid, yielding 0.18 g of the target compound. The yield was 51.9%, and the purity was 97.9%. MS m / z (ESI): 424.3 [M+1]. 1 HNMR(400MHz, CDCl3)δ9.90(s,1H),8.27(d,J=8.0Hz,1H),7.84(s,1H),7.75(t,J =7.6Hz,1H),7.65(d,J=8.2Hz,1H),7.47(t,J=7.5Hz,1H),7.03(dd,J=25.7,8.1H z,2H),6.16(s,1H),4.19(d,J=5.2Hz,2H),3.80(s,2H),3.39(d,J=10.8Hz,2H),2 .99(d,J=10.8Hz,2H),2.63(d,J=6.9Hz,1H),2.18(s,3H),1.90(d,J=8.1Hz,1H).

[0179] Example 6

[0180] Preparation of N-(5-chloro-2-methylphenyl)-2-methyl-4-((4-oxo-3,4-dihydroquinazolin-2-yl)methyl)piperazine-1-carboxamide:

[0181]

[0182] Preparation route:

[0183]

[0184] Step 1: Synthesis of 2-(chloromethyl)quinazolin-4-(3H)-quinazolinone, the specific process is the same as step 1 in Example 1.

[0185] Step 2: Synthesis of tert-butyl 2-methyl-4-((4-oxo-3,4-dihydroquinazolin-2-yl)methyl)piperazine-1-carboxylic acid:

[0186]

[0187] At room temperature, 2-chloromethyl-4-(3H)-quinazolinone (1 eq), 2-methylpiperazine-1-carboxylic acid tert-butyl ester (2 eq), and potassium carbonate (3 eq) were placed in a reaction tube. Then, dichloromethane was added, and the mixture was stirred at room temperature for 12 h. After the reaction was complete, water was added to quench the reaction, and the mixture was extracted with ethyl acetate. The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and column chromatography was performed to obtain 6.12 g of the target product as a white solid with a yield of 59.6% and a purity of 98.1%. MS m / z (ESI): 359.3 [M+1].

[0188] Step 3: Synthesis of 2-((3,6-diazabicyclo[3.1.1]heptane-3-yl)methyl)-4(3H)-quinazolinone:

[0189]

[0190] At room temperature, 2-methyl-4-((4-oxo-3,4-dihydroquinazolin-2-yl)methyl)piperazine-1-carboxylic acid tert-butyl ester (1 eq) was placed in a reaction tube, followed by the addition of dichloromethane and then a 4N hydrogen chloride-dioxane solution. The mixture was stirred at room temperature until the reaction was complete. Sodium hydroxide was added dropwise until the pH reached 8-10. The solid was filtered and dried to obtain 2.3 g of the target product as a white solid with a yield of 62.2% and a purity of 97.9%. MS m / z (ESI): 275.3 [M+1].

[0191] Step 4: Synthesis of N-(5-chloro-2-methylphenyl)-2-methyl-4-((4-oxo-3,4-dihydroquinazolin-2-yl)methyl)piperazine-1-carboxamide:

[0192]

[0193] At room temperature, 2-((3,6-diazabicyclo[3.1.1]heptane-3-yl)methyl)-4(3H)-quinazolinone (1 eq), 5-chloro-2-methylphenyl isocyanate (2 eq), and triethylamine (3 eq) were placed in a reaction tube, followed by the addition of tetrahydrofuran. The mixture was stirred at room temperature for 12 h until the reaction was complete. The mixture was then extracted with dichloromethane and water. The organic phases were combined, and the final product was obtained by column chromatography as a white solid, yielding 0.21 g of the target compound. The yield was 63.3%, and the purity was 97.4%. MS m / z (ESI): 426.3 [M+1]. 1 HNMR (400MHz, CDCl3) δ10.03 (s, 1H), 8.28 (d, J = 7.9Hz, 1H), 7.78 (t, J = 7.6Hz, 1H), 7.67 (d,J=9.2Hz,2H),7.49(t,J=7.5Hz,1H),7.07(d,J=8.1Hz,1H),6.99(d,J=8.1Hz,1H),6. 22(s,1H),3.98–3.68(m,3H),3.54–3.30(m,2H),3.04(dd,J=12.5,9.1Hz,1H),2.86(d, J=11.8Hz,1H),2.72(s,1H),2.56(t,J=10.5Hz,1H),2.20(s,3H),1.14(d,J=6.1Hz,3H).

[0194] Example 7

[0195] Preparation of (1S,4S)-N-(5-chloro-2-methylphenyl)-5-((4-oxo-3,4-dihydroquinazolin-2-yl)methyl)-2,5-diazabicyclo[2.2.1]heptane-2-carboxamide:

[0196]

[0197] Preparation route:

[0198]

[0199] Step 1: Synthesis of 2-(chloromethyl)quinazolin-4-(3H)-quinazolinone, the specific process is the same as step 1 in Example 1.

[0200] Step 2: Synthesis of (1S,4S)-5-((4-oxo-3,4-dihydroquinazolin-2-yl)methyl)-2,5-diazabicyclo[2.2.1]heptane-2-carboxylic acid tert-butyl ester:

[0201]

[0202] At room temperature, 2-chloromethyl-4-(3H)-quinazolinone (1 eq), (1S,4S)-2-Boc-2,5-diazabicyclo[2.2.1]heptane (2 eq), and potassium carbonate (3 eq) were placed in a reaction tube. Then, dichloromethane was added, and the mixture was stirred at room temperature for 12 h. After the reaction was complete, water was added to quench the reaction, and the mixture was extracted with ethyl acetate. The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and column chromatography was performed to obtain 5.98 g of the target product as a white solid with a yield of 53.1% and a purity of 97.1%. MS m / z (ESI): 357.3 [M+1].

[0203] Step 3: Synthesis of 2-(((1S,4S)-2,5-diazabicyclo[2.2.1]heptane-2-yl)methyl)-4(3H)-quinazolinone:

[0204] At room temperature, (1S,4S)-5-((4-oxo-3,4-dihydroquinazolin-2-yl)methyl)-2,5-diazabis

[0205]

[0206] [2.2.1]Heptane-2-carboxylic acid tert-butyl ester (1 eq) was placed in a reaction tube, followed by the addition of dichloromethane and then 4N hydrogen chloride-dioxane solution. The mixture was stirred at room temperature until the reaction was complete. Sodium hydroxide was added dropwise until the pH reached 8-10. The solid was filtered and dried to obtain 2.2 g of the target product, a white solid with a yield of 57.9% and a purity of 96.8%. MS m / z (ESI): 257.2 [M+1].

[0207] Step 4: Synthesis of (1S,4S)-N-(5-chloro-2-methylphenyl)-5-((4-oxo-3,4-dihydroquinazolin-2-yl)methyl)-2,5-diazabicyclo[2.2.1]heptane-2-carboxamide:

[0208]

[0209] At room temperature, 2-(((1S,4S)-2,5-diazabicyclo[2.2.1]heptane-2-yl)methyl)-4(3H)-quinazolinone (1 eq), 5-chloro-2-methylphenyl isocyanate (2 eq), and triethylamine (3 eq) were placed in a reaction tube, followed by the addition of tetrahydrofuran. The mixture was stirred at room temperature for 12 h until the reaction was complete. The mixture was then extracted with dichloromethane and water. The organic phases were combined, and the final product was obtained by column chromatography as a white solid, yielding 0.22 g of the target compound. The yield was 65.6%, and the purity was 98.3%. MS m / z (ESI): 424.2 [M+1]. 1H NMR (400MHz, CDCl3) δ10.09(s,1H),8.27(d,J=7.9Hz,1H),7.91(s,1H),7.76(t,J=7.6Hz, 1H),7.65(d,J=8.1Hz,1H),7.48(t,J=7.5Hz,1H),7.06(d,J=8.1Hz,1H),6.97(d,J=8.1Hz, 1H),5.99(s,1H),4.65(s,1H),3.84(q,J=15.9Hz,2H),3.70–3.57(m,2H),3.44(d,J=8.4H z, 1H), 2.99 (q, J = 9.7Hz, 2H), 2.21 (s, 3H), 2.02 (d, J = 10.0Hz, 1H), 1.90 (d, J = 10.0Hz, 1H).

[0210] Example 8

[0211] Preparation of N-(5-chloro-2-methylphenyl)-4-((4-oxo-3,4-dihydroquinazolin-2-yl)methyl)-1,4-diazacycloheptane-1-carboxamide:

[0212]

[0213] Preparation route:

[0214]

[0215] Step 1: Synthesis of 2-(chloromethyl)quinazolin-4-(3H)-quinazolinone, the specific process is the same as step 1 in Example 1.

[0216] Step 2: Synthesis of tert-butyl 4-((4-oxo-3,4-dihydroquinazolin-2-yl)methyl)-1,4-diazazo-1-carboxylic acid:

[0217]

[0218] At room temperature, 2-chloromethyl-4-(3H)-quinazolinone (1 eq), tert-butyl 1,4-diazacycloheptane-1-carboxylate (2 eq), and potassium carbonate (3 eq) were placed in a reaction tube. Then, dichloromethane was added, and the mixture was stirred at room temperature for 12 h. After the reaction was complete, water was added to quench the reaction, and the mixture was extracted with ethyl acetate. The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and column chromatography was performed to obtain 5.88 g of the target product as a white solid with a yield of 51.3% and a purity of 96.2%. MS m / z (ESI): 359.3 [M+1].

[0219] Step 3: Synthesis of 2-((1,4-diazacycloheptane-1-yl)methyl)-4(3H)-quinazolinone:

[0220]

[0221] At room temperature, 4-((4-oxo-3,4-dihydroquinazolin-2-yl)methyl)-1,4-diazaphen-1-carboxylic acid tert-butyl ester (1 eq) was placed in a reaction tube, followed by the addition of dichloromethane and then a 4N hydrogen chloride-dioxane solution. The mixture was stirred at room temperature until the reaction was complete. Sodium hydroxide was added dropwise until the pH reached 8-10. The solid was filtered and dried to obtain 2.1 g of the target product as a white solid with a yield of 54.6% and a purity of 97.2%. MS m / z (ESI): 245.3 [M+1].

[0222] Step 4: Synthesis of N-(5-chloro-2-methylphenyl)-4-((4-oxo-3,4-dihydroquinazolin-2-yl)methyl)-1,4-diazacycloheptane-1-carboxamide:

[0223]

[0224] At room temperature, 2-((1,4-diazacycloheptane-1-yl)methyl)-4(3H)-quinazolinone (1 eq), 5-chloro-2-methylphenyl isocyanate (2 eq), and triethylamine (3 eq) were placed in a reaction tube, followed by the addition of tetrahydrofuran. The mixture was stirred at room temperature for 12 h until the reaction was complete. The mixture was then extracted with dichloromethane and water. The organic phases were combined, and the final product was obtained by column chromatography as a white solid, yielding 0.23 g of the target compound. The yield was 68.1%, and the purity was 97.3%. MS m / z (ESI): 426.3 [M+1]. 1 H NMR (400MHz, CDCl3) δ10.11(s,1H),8.28(d,J=7.9Hz,1H),7.84(s,1H),7.77(t,J=7.6Hz,1H),7.66(d,J=8.1Hz,1H),7.49(t,J=7.4Hz,1H),7. 06(d,J=8.0Hz,1H),6.97(d,J=8.1Hz,1H),6.22(s,1H),3.79–3.52(m,6H),2.96–2.71(m,4H),2.14(d,J=47.4Hz,3H),2.00(d,J=38.7Hz,2H).

[0225] Example 9

[0226] Preparation of N-(5-chloro-2-methylphenyl)-4-((4-oxo-3,4-dihydroquinazolin-2-yl)methyl)piperazine-1-thioamide:

[0227]

[0228] Preparation route:

[0229]

[0230] Step 1: Synthesis of 2-(chloromethyl)quinazolin-4-(3H)-quinazolinone, the specific process is the same as step 1 in Example 1.

[0231] Step 2: Synthesis of 4-((4-oxo-3,4-dihydroquinazolin-2-yl)methyl)piperazine-1-carboxylic acid tert-butyl ester, the specific process is the same as step 2 of Example 1.

[0232] Step 3: Synthesis of 2-(piperazine-1-methyl)quinazolin-4(3H)-one, the specific process is the same as step 3 of Example 1.

[0233] Step 4: Synthesis of N-(5-chloro-2-methylphenyl)-4-((4-oxo-3,4-dihydroquinazolin-2-yl)methyl)piperazine-1-thioamide:

[0234]

[0235] At room temperature, 2-(piperazin-1-methyl)-4-(3H)-quinazolinone (1 eq), 5-chloro-2-methylphenyl isothiocyanate (2 eq), and triethylamine (3 eq) were placed in a reaction tube, followed by the addition of tetrahydrofuran. The mixture was stirred at room temperature for 12 h until the reaction was complete. The mixture was then extracted with dichloromethane and water. The organic phases were combined, and the final product was obtained by column chromatography as a white solid, yielding 0.19 g of the target compound. The yield was 56.8%, and the purity was 98.2%. MS m / z (ESI): 428.2 [M+1]. 1 H NMR (400MHz, CDCl3) δ9.98(s,1H),8.29(d,J=7.9Hz,1H),7.88(d,J=7.7Hz,1H),7.78(t,J=7.6Hz,1H),7.68(d,J=8.1Hz,1H),7. 50(t,J=7.5Hz,1H),6.93(t,J=9.6Hz,1H),6.77(d,J=5.4Hz,1H),6.55(s,1H),3.63(d,J=12.2Hz,6H),2.68(s,4H),2.31(s,3H).

[0236] Example 10

[0237] Preparation of N-(5-chloro-2-methylphenyl)-3-((7-fluoro-4-oxo-3,4-dihydroquinazolin-2-yl)methyl)-3,6-diazabicyclo[3.1.1]heptane-6-carboxamide:

[0238]

[0239] Preparation route:

[0240]

[0241] Step 1: Synthesis of 2-chloromethyl-7-fluoro-4(3H)-quinazolinone:

[0242]

[0243] At room temperature, methyl 2-amino-4-fluorobenzoate (1 eq) and chloroacetonitrile (3 eq) were placed in a reaction tube, followed by the addition of a 4N hydrogen chloride-dioxane solution, and the temperature was raised to 80 °C. After reacting for 24 hours, the reaction solution was cooled to room temperature, the resulting solid was collected and dissolved in water, neutralized with sodium hydroxide to pH = 7, the solid was collected by filtration, washed with water and dried, finally yielding 6.33 g of the target product, a white solid, with a yield of 62.2%, purity of 96.1%, and MS m / z (ESI): 212.1 [M+1].

[0244] Step 2: Synthesis of tert-butyl 3-((7-fluoro-4-oxo-3,4-dihydroquinazolin-2-yl)methyl)-3,6-diazabicyclo[3.1.1]heptane-6-carboxylic acid:

[0245]

[0246] At room temperature, 2-chloromethyl-7-fluoro-4(3H)-quinazolinone (1 eq), 6-N-Boc-3,6-diazabicyclo[3.1.1]heptane (2 eq), and potassium carbonate (3 eq) were placed in a reaction tube. Then, dichloroethane was added, and the mixture was stirred at room temperature for 12 h. After the reaction was complete, water was added to quench the reaction, and the mixture was extracted with ethyl acetate. The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and column chromatography was performed to obtain 6.12 g of the target product as a white solid with a yield of 57.7% and a purity of 97.1%. MS m / z (ESI): 375.3 [M+1].

[0247] Step 3: Synthesis of 2-((3,6-diazabicyclo[3.1.1]heptane-3-yl)methyl)-7-fluoro-4(3H)-quinazolinone:

[0248] At room temperature, 3-((7-fluoro-4-oxo-3,4-dihydroquinazolin-2-yl)methyl)-3,6-diazabicyclo[3.1.1]heptane-6-carboxylic acid tert-butyl ester (1 eq) was placed in a reaction tube, followed by the addition of dichloromethane and then a 4N hydrogen chloride-dioxane solution. The mixture was stirred at room temperature until the reaction was complete. Sodium hydroxide was added dropwise until the pH reached 8-10. The solid was filtered and dried to obtain 2.3 g of the target product as a white solid with a yield of 63.3%, a purity of 98.1%, and MS m / z (ESI): 275.2 [M+1].

[0249]

[0250] Step 4: Synthesis of N-(5-chloro-2-methylphenyl)-3-((7-fluoro-4-oxo-3,4-dihydroquinazolin-2-yl)methyl)-3,6-diazabicyclo[3.1.1]heptane-6-carboxamide:

[0251]

[0252] At room temperature, 2-((3,6-diazabicyclo[3.1.1]heptane-3-yl)methyl)-7-fluoro-4(3H)-quinazolinone (1 eq), 5-chloro-2-methylphenyl isocyanate (2 eq), and triethylamine (3 eq) were placed in a reaction tube, followed by the addition of tetrahydrofuran. The mixture was stirred at room temperature for 12 h until the reaction was complete. The mixture was then extracted with dichloromethane and water. The organic phases were combined, and the final product was obtained by column chromatography as a white solid, yielding 0.19 g of the target compound. The yield was 47.7%, and the purity was 96.3%. MS m / z (ESI): 442.2 [M+1]. 1 HNMR (400MHz, CDCl3) δ9.90 (s, 1H), 8.27 (d, J = 8.0Hz, 1H), 7.84 (s, 1H), 7.65 (d, J = 8.2Hz, 1H), 7.47 (t, J = 7.5Hz, 1H), 7.03 (dd, J = 25.7, 8.1Hz, 2H), 6.1 1(s,1H),4.19(d,J=5.2Hz,2H),3.80(s,2H),3.40(d,J=10.8Hz,2H),2.99( d,J=10.8Hz,2H),2.63(d,J=6.9Hz,1H),2.18(s,3H),1.90(d,J=8.1Hz,1H).

[0253] Cell proliferation inhibition experiment:

[0254] HCT-15BRCA2(- / -) cell lines, Capan-1BRCA2(- / -) cell lines, HCC1937BRCA1(- / -) cell lines, and BRCA-free cell lines 4T1, MDA-MB-231, and MDA-MB-468 were all purchased from ATCC and cultured in RPMI 1640 containing 10% fetal bovine serum (FBS) and 1× penicillin-streptomycin (all three were purchased from Gibco).

[0255] The following section uses the HCT-15BRCA2(- / -) cell line as an example to introduce the specific procedures for cell proliferation inhibition experiments:

[0256] (1) 3000 HCT-15BRCA2(- / -) cells were added to each well of a 96-well plate (purchased from PerkinElmer) and cultured at 37°C and 5% CO2 for 12 hours.

[0257] (2) The compounds prepared in Examples 1 to 10 were dissolved in DMSO, then added to the culture medium, and then added to 96-well plates to make the concentration of the compound in the wells 6 μmol. The plates were then cultured at 37°C and 5% CO2 for 3 days.

[0258] (3) After 3 days, the culture plate was removed and placed at room temperature for 10 minutes. Then, 20 μl of MTT reagent (purchased from Beyotime) was added and cultured at 37°C and 5% CO2 for 4 hours. The supernatant was discarded, and 150 μl of DMSO was added to each well. The plate was shaken at low speed for 10 minutes to fully dissolve the crystals. The absorbance of each well was measured at OD490 nm using an enzyme-linked immunosorbent assay (ELISA) reader. The inhibition rate of each compound was calculated.

[0259] The experimental procedures for the other cell lines were the same as those for the HCT-15BRCA2(- / -) cell line. In the cell proliferation inhibition experiments of the non-BRCA mutant cell lines 4T1, MDA-MB-231, and MDA-MB-468, the concentration of the compound in the well plate was 25 μmol. In the cell proliferation inhibition experiments of the Capan-1BRCA2(- / -) cell line and the HCC1937BRCA1(- / -) cell line, the concentration of the compound in the well plate was 6 μmol.

[0260] The cell proliferation inhibition effects of the compounds olaparib and AZD5305, which are already on the market, are compared in Table 1:

[0261] Table 1. Cell proliferation inhibition effects of different compounds

[0262]

[0263] As shown in Table 1, the compounds prepared in Examples 1-10 exhibit good selectivity for both BRCA-mutated and non-mutated cells. In particular, they show excellent inhibition rates against BRCA-mutated cells at low concentrations. At a concentration of only 6 μmol, the proliferation inhibition rate against BRCA-mutated cell lines reached over 70%, which is significantly higher than the proliferation inhibition rate against non-BRCA-mutated cell lines at a higher concentration of 25 μmol. This indicates that the drug demonstrates a higher proliferation inhibition rate against BRCA-mutated cell lines, and is superior to olaparib and AZD5305. Specifically, the compounds prepared in Examples 1 and 10 showed a cell proliferation inhibition rate of over 66% against the HCC1937BRCA1(- / -) cell line, the compounds prepared in Examples 2, 4, and 5 all showed a cell proliferation inhibition rate of over 70% against the HCT-15BRCA2(- / -) cell line, and the compounds prepared in Examples 1, 5, and 10 all showed a cell proliferation inhibition rate of over 50% against the Capan-1BRCA2(- / -) cell line, which is significantly better than the cell proliferation inhibition effect of existing compounds.

[0264] Tumor formation experiment in nude mice:

[0265] The compound N-(5-chloro-2-methylphenyl)-3-((4-oxo-3,4-dihydroquinazolin-2-yl)methyl)-3,6-diazabicyclo[3.1.1]heptane-6-carboxamide prepared in Example 5 was used in a tumorigenesis experiment in nude mice, as follows:

[0266] HCT-15 cells (5 × 10⁶) were injected subcutaneously into the tissue of 5-week-old female BALB / c nude mice. When the tumor volume reached 50 mm², the cells were injected into the tumor tissue. 3 At approximately 14 days, mice were randomly divided into two groups (n=6 per group) and injected intraperitoneally with a dose of 10 mg / kg every two days for 14 consecutive days. The tumor volume was measured every 2 days and calculated using the formula: length × width 2 / 2. The weight of the mice was also recorded. At the terminal stage, all mice were sacrificed, and the tumors were isolated and weighed.

[0267] like Figures 1-3 As shown, 4 days after the start of administration, the relative tumor volume of the mice in the administration group was significantly smaller than that in the control group. One week after the start of administration, the body weight of the mice in the administration group was significantly lower than that in the control group. Furthermore, the difference in relative tumor volume between the mice in the administration group and the control group increased continuously with the increase of administration time, and the tumor weight also decreased significantly. This confirms that the drug can effectively inhibit tumor growth in mice at low doses, and the body weight of the mice did not change significantly, indicating that there were no obvious toxic side effects at this dose.

[0268] In summary, this invention provides a selective inhibitor against BRCA mutations. This selective inhibitor can effectively inhibit the proliferation of tumor cells with BRCA mutations at low dosage concentrations without significant toxic side effects. It can be used to treat diseases related to BRCA mutations, such as breast cancer, ovarian cancer, lung cancer, pancreatic cancer, gastric cancer, breast cancer, brain tumors, melanoma, renal cell carcinoma, colorectal cancer, head and neck cancer, bladder cancer, prostate cancer, hepatocellular carcinoma, or cholangiocarcinoma.

[0269] The above description is merely a preferred embodiment of the present invention and is not intended to limit the invention. Various modifications and variations can be made to the present invention by those skilled in the art. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the scope of protection of the present invention.

Claims

1. A compound or a pharmaceutically acceptable salt thereof, characterized in that, The compound is one of the following compounds: 。 2. A compound represented by formula (II) below, or a pharmaceutically acceptable salt thereof, （Ⅱ）， in, R2 is selected from ， 。 3.N-(5-chloro-2-methylphenyl)-3-((4-oxo-3,4-dihydroquinazolin-2-yl)methyl)-3,6-diazabicyclo[3.1.1]heptane-6-carboxamide. 4.N-(5-chloro-2-methylphenyl)-4-((4-oxo-3,4-dihydroquinazolin-2-yl)methyl)-1,4-diazacycloheptane-1-carboxamide.

5. A pharmaceutical composition, characterized in that, It comprises the compound as described in claim 1 or 2 or a pharmaceutically acceptable salt thereof, and at least one pharmaceutically acceptable excipient.

6. Use of the compound of claim 1 or 2 or a pharmaceutically acceptable salt thereof in the preparation of a medicament for treating BRCA-mutated cancers; The cancers mentioned are selected from breast cancer, ovarian cancer, lung cancer, pancreatic cancer, gastric cancer, melanoma, renal cell carcinoma, colorectal cancer, head and neck cancer, bladder cancer, prostate cancer, hepatocellular carcinoma, or bile duct cancer.

Images