A quinazoline compound, a preparation method thereof and application thereof in preparation of PRMT5 / EGFR antitumor inhibitor drugs

Abstract

The application provides a quinazoline compound, a preparation method of the quinazoline compound and application of the quinazoline compound in preparation of a PRMT5 / EGFR antitumor inhibitor drug, and belongs to the technical field of medicines.The application introduces a quinazoline mother nucleus into the design of a PRMT5 inhibitor for the first time through a pharmacophore fusion strategy, and the designed and synthesized compound has strong PRMT5 / EGFR double-target point inhibition capability.The application introduces a quinazoline mother nucleus into the design of a PRMT5 inhibitor for the first time, and the quinazoline compound can be used as an active ingredient to prepare a therapeutic drug for diseases caused by abnormal activation of epigenetic enzymes PRMT5 and EGFR kinases, and also shows potential application value in preparation of a drug for treating and / or preventing proliferative diseases and cancers.

Description

TECHNICAL FIELD

[0001] The application belongs to the technical field of medicine, and particularly relates to a quinazoline compound, a preparation method thereof and application of the quinazoline compound in preparation of a PRMT5 / EGFR antitumor inhibitor drug. BACKGROUND

[0002] Breast cancer and lung cancer are two major malignant tumors with high morbidity and mortality, which pose a serious threat to human life and health. According to the global cancer statistics data released by the International Agency for Research on Cancer, there were about 19.96 million new cancer cases and about 9.74 million deaths worldwide in 2022. At present, the number of new cancer cases worldwide each year is close to 19 million, and the number of deaths is about 10 million; if effective prevention and control interventions cannot be implemented, it is estimated that by 2050, the number of new cases and deaths will rise to 33 million and 18 million, respectively. Although in recent years, under the continuous exploration of scientific researchers, the comprehensive treatment strategies such as surgery, radiotherapy, chemotherapy and immunotherapy have made great progress, but the problems of tumor heterogeneity, drug resistance and metastasis and recurrence are still the core bottlenecks that limit the further improvement of clinical efficacy. Therefore, further elucidating the molecular mechanisms of tumor occurrence and development, exploring new therapeutic targets, and developing efficient and low-drug-resistance antitumor drugs are still important research directions in the field of antitumor drug research at the present stage.

[0003] In the process of tumor occurrence and development, in addition to the classic gene mutation and chromosomal abnormalities, epigenetic abnormalities also play a crucial role. Epigenetic regulation mainly affects gene expression without changing the DNA sequence through DNA methylation, histone modification, chromatin remodeling and non-coding RNA regulation. Among them, histone methylation modification is catalyzed by protein arginine methyltransferase family and lysine methyltransferase family, and is widely involved in biological processes such as gene transcription, DNA damage repair, RNA processing and cell cycle regulation. Disorders of epigenetic regulation can lead to abnormal activation of oncogenes or silencing of tumor suppressor genes, thereby driving tumor occurrence and progression. In recent years, targeting epigenetic regulatory enzymes has become a hot field of anti-tumor drug research, and several targets such as EZH2, LSD1, HDAC have successfully developed into marketed drugs.

[0004] Arginine methyltransferase 5 (PRMT5), a typical representative of the type II arginine methyltransferase family, has the core function of catalyzing the symmetrical dimethylation of arginine residues in histones H2A, H3, H4, and various non-histone molecules. PRMT5 needs to bind with methylsomal protein 50 (MEP50) to form a stable heterooctamer complex, thereby participating in the regulation of several key life activities, including gene transcription, mRNA splicing, DNA damage repair, signal transduction, and cell proliferation and differentiation. Numerous studies have shown that PRMT5 is highly expressed in lung cancer, breast cancer, colorectal cancer, glioblastoma, melanoma, and hematologic malignancies, and its expression level is closely related to poor patient prognosis. Abnormal activation of PRMT5 can drive malignant tumor progression through multiple pathways, including promoting oncogene expression, maintaining tumor stem cell characteristics, and inhibiting apoptosis signaling.

[0005] Notably, PRMT5 also exhibits potential as a synthetic lethal target. Studies have shown that PRMT5 inhibition can selectively induce tumor cell death in tumor cells lacking methionine phosphorylase, providing a new strategy for developing highly selective and low-toxicity PRMT5 inhibitors. Based on this, PRMT5 has become one of the most promising targets in the development of epigenetic anti-tumor drugs.

[0006] Epidermal growth factor receptor (EGFR) is one of the most classic targets for lung and breast cancer, and the development of inhibitors has undergone several generations of updates, aiming to address drug resistance caused by mutations. For example, first- and second-generation EGFR inhibitors, such as Gefitinib, Erlotinib, and Afatinib, which use quinazoline as a backbone, primarily target metastatic non-small cell lung cancer (NSCLC) with EGFR exon 19 deletion or exon 21 (L858R) mutations. However, drug resistance caused by the T790M mutation led to the development of third-generation EGFR inhibitors such as Osimertinib. Currently, drug resistance caused by the C797S mutation is forcing the development of fourth-generation EGFR inhibitors. The C797S mutation results in the absence of amino acid residues that can form covalent bonds in the inhibitor; therefore, fourth-generation EGFR inhibitors will utilize hydrophobic interactions and hydrogen bonding to stably bind to the active site of EGFR.

[0007] The development of dual-target inhibitors is one strategy to address drug resistance, as some studies have shown that PRMT5 can regulate the NF-κB pathway by modulating the EGFR cascade amplification effect. Therefore, this "upstream and downstream" relationship is crucial for the design of dual-target inhibitors. Current research on PRMT5 inhibitors mostly uses a 7-denitrile nucleus structure, with a core pentose sugar structure in the middle, and an aromatic group on the far right forming a hydrophobic interaction with the hydrophobic residue Phe327. While they exhibit good activity, nucleoside inhibitors have potential serious adverse reactions and pharmacokinetic problems. For example, the inherent pharmacokinetic defects and safety issues of nucleoside inhibitors are becoming increasingly prominent; these molecules are highly polar, have poor lipid solubility, and generally have low oral bioavailability; their activity depends on intracellular three-step phosphorylation activation, making them susceptible to the influence of kinase expression levels and metabolic enzymes, resulting in short half-lives and significant individual variability. More importantly, these compounds are easily recognized by mitochondrial DNA polymerase γ, causing mitochondrial toxicity, clinically manifested as lactic acidosis, lipemia, bone marrow suppression, and renal tubular damage, among other serious adverse reactions. Therefore, developing novel non-nucleoside PRMT5 inhibitors is of great significance.

[0008] Due to the potential benefits of PRMT5 in cancer treatment, but with no FDA-approved drugs yet available, it remains in the clinical stage. The development of PRMT5 inhibitors has attracted significant interest from academic and industrial pharmaceutical researchers, including substrate-competitive inhibitors (such as GSK3326595), SAM-competitive inhibitors (such as Onametostat (JNJ64619178)), and MTA cooperation inhibitors (such as MRTX-1719). Furthermore, a series of non-nucleoside derivatives have been reported as SAM-competitive PRMT5 inhibitors, as well as some interesting bioactive inhibitors that bind to allosteric or adapter binding sites. Since methyltransferases utilize SAM molecules as methyl donors for their catalytic activity, discovering novel SAM-competitive inhibitors is not only important for PRMT5 but also provides new strategies for developing other PRMT inhibitors. Therefore, based on the solved co-crystal structure of PRMT5 and SAM-competitive inhibitors, this invention initiates a design and synthesis process for discovering novel SAM-competitive PRMT5 inhibitors, hoping to also find a new scaffold for developing other methyltransferase inhibitors. Summary of the Invention

[0009] To develop novel and highly effective PRMT5 / EGFR antitumor inhibitors, this invention proposes a quinazoline-containing compound, its preparation method, and its application in the preparation of PRMT5 / EGFR antitumor inhibitor drugs. This invention has conducted extensive research on quinazoline-containing compounds, continuously altering the structure of the compounds through molecular docking results and cellular and epigenetic enzyme inhibitory activities, and designing and synthesizing a series of novel quinazoline-containing compounds.

[0010] To achieve the above objectives, the present invention provides the following technical solution: This invention provides a quinazoline-containing compound with the structural formula shown in Formula I: ; In Equation I, R2 is selected from , , , , , , , , , and One of them; When R2 is selected or When, R1 is selected ; When R2 is selected , , , , , , and When one of them is selected, R1 is chosen from ; When R2 is selected When, R1 is selected , , , , , , , , , , , , , , , , , and One of them; Where R1 is selected At that time, the structural formula of the quinazoline-containing compound is shown in Formula II: In Formula II, R3 is selected from , , , , and One of them; When R1 is selected At that time, the structural formula of the quinazoline-containing compound is shown in Formula III: R4 is selected from and One of them; When R1 is selected At that time, the structural formula of the quinazoline-containing compound is shown in Formula IV: R5 is selected from , , , , , , , , and One of them.

[0011] Furthermore, the quinazoline-containing compound includes: N -(4-morpholinoquinazolin-7-yl)-3-phenylpropionamide, N -(4-morpholinoquinazolin-7-yl)-3-(p-tolyl)propionamide, N -(4-morpholinoquinazolin-7-yl)-3-(4-trifluoromethylphenyl)propionamide, N -(4-morpholinoquinazolin-7-yl)-4-phenylbutyramide, N -(4-morpholinoquinazolin-7-yl)-4-(p-tolyl)butyramide, N -(4-morpholinoquinazolin-7-yl)-5-phenylpentanamide, 4-(4-bromophenyl)- N -(4-morpholinylquinazolin-7-yl)butyramide, 4-(4-chlorophenyl)- N -(4-morpholinylquinazolin-7-yl)butyramide, 4-(4-aminophenyl)- N -(4-morpholinylquinazolin-7-yl)butyramide, 4-(4-fluorophenyl)- N -(4-morpholinoquinazolin-7-yl)butyramide, N -(4-((2-aminoethyl)amino)quinazolin-7-yl)-4-(p-tolyl)butyramide, N -(4-(piperazin-1-yl)quinazolin-7-yl)-4-(p-tolyl)butyramide, N-(4-morpholinylquinazoline-7-yl)bicyclo[4.2.0]oct-1(6),2,4-trien-7-carboxamide, N -(4-morpholinoquinazolin-7-yl)-3-(1-naphthyl)propionamide, N 1 -(4-morpholinoquinazolin-7-yl)- N 2 -Phenylacetamide, N -(4-morpholinoquinazolin-7-yl)-1-phenylmethanesulfonamide, 1-benzoyl- N -(4-morpholinoquinazolin-7-yl)piperidine-4-carboxamide, 1-(4-fluorobenzoyl)- N -(4-morpholinoquinazolin-7-yl)piperidine-4-carboxamide, 1-(4-methoxybenzoyl)- N -(4-morpholinoquinazolin-7-yl)piperidine-4-carboxamide, 1-(4-methylbenzoyl)- N -(4-morpholinoquinazolin-7-yl)piperidine-4-carboxamide, 1-(2-methylbenzoyl)- N -(4-morpholinoquinazolin-7-yl)piperidine-4-carboxamide, N -(4-morpholinoquinazolin-7-yl)-1-(3-trifluoromethylbenzoyl)piperidine-4-carboxamide, 2-(4-(2-methylbenzoyl)piperazin-1-yl)- N -(4-morpholinoquinazolin-7-yl)acetamide, 2-(4-(4-methylbenzoyl)piperazin-1-yl)- N -(4-morpholinoquinazolin-7-yl)acetamide, 1-(benzylsulfonyl)- N -(4-morpholinoquinazolin-7-yl)pyrrolidine-2-carboxamide, N -(4-morpholinoquinazolin-7-yl)-1-(3-trifluoromethylbenzoyl)pyrrolidine-2-carboxamide, N -(4-morpholinoquinazolin-7-yl)-1-(2-trifluoromethylbenzoyl)pyrrolidine-2-carboxamide, 1-(3,5-bis(trifluoromethyl)benzoyl)- N -(4-morpholinoquinazolin-7-yl)pyrrolidine-2-carboxamide, 1-(cyclohexaneformyl)- N -(4-morpholinoquinazolin-7-yl)pyrrolidine-2-carboxamide, 1-(9,10-dioxo-9,10-dihydroanthracene-2-formyl)- N -(4-morpholinoquinazolin-7-yl)pyrrolidine-2-carboxamide, 1-((4-methoxyphenyl)sulfonyl)- N-(4-morpholinoquinazolin-7-yl)pyrrolidine-2-carboxamide, 1-((5-(dimethylamino)naphthyl-1-yl)sulfonyl)- N -(4-morpholinoquinazolin-7-yl)pyrrolidine-2-carboxamide, N -(4-morpholinoquinazolin-7-yl)-1-(1-phenethyl)pyrrolidine-2-carboxamide, 1-(4-methylbenzyl)- N -(4-morpholinoquinazolin-7-yl)pyrrolidine-2-carboxamide, N -(4-Fluorophenyl)- N -(4-morpholinoquinazolin-7-yl)cyclopropane-1,1-dicarboxamide N -(5-benzyloxypyridin-2-yl)-4-morpholinoquinazoline-7-amine N 4 -(2-aminoethyl)- N 7 -(5-benzyloxypyridin-2-yl)quinazolin-4,7-diamine N -(5-benzyloxypyridin-2-yl)-4-(piperazin-1-yl)quinazolin-7-amine N -(5-benzyloxypyridin-2-yl)-4-(4-methylpiperazin-1-yl)quinazolin-7-amine N -(5-benzyloxypyridin-2-yl)-4-(4-cyclopropylpiperazin-1-yl)quinazolin-7-amine N 7 -(5-Benzyloxypyridin-2-yl)- N 4 , N 4 -Diethylquinazolin-4,7-diamine N -(5-benzyloxypyridin-2-yl)-4-((2S,6R)-2,6-dimethylmorpholino)quinazolin-7-amine, N -(5-benzyloxypyridin-2-yl)-4-(4-methanesulfonylpiperazin-1-yl)quinazolin-7-amine and 1-(4-(7-(((5-benzyloxypyridin-2-yl)amino)quinazolin-4-yl)piperazin-1-yl)ethyl-1-one.

[0012] The present invention also provides a method for preparing the above-mentioned quinazoline-containing compounds, using 4-chloro-7-nitroquinazoline as the starting material, introducing a nitrogen-containing substituent at the 4-position of the quinazoline ring through a nucleophilic substitution reaction to obtain a 4-substituted-7-nitroquinazoline intermediate; then reducing the 7-position nitro group to an amino group through a reduction reaction to obtain a 4-substituted-7-aminoquinazoline intermediate; finally, preparing the target compound through amidation, sulfonation, alkylation or Buchwald-Hartwig coupling reaction.

[0013] Furthermore, the preparation method of the quinazoline-containing compound in this invention includes the following steps: (1) Using 4-chloro-7-nitroquinazoline as the starting material, a nucleophilic substitution reaction was carried out with a nitrogen-containing nucleophilic reagent in an organic solvent in the presence of a base to prepare a 4-substituted-7-nitroquinazoline intermediate; (2) The intermediate obtained in step (1) is reduced in a reduction system to reduce the 7-nitro group to an amino group to obtain a 4-substituted-7-aminoquinazoline intermediate; (3) The intermediate obtained in step (2) is subjected to amidation condensation reaction, acylation reaction, sulfonation reaction, alkylation reaction or Buchwald-Hartwig coupling reaction to obtain the target compound.

[0014] Furthermore, in step (1), the organic solvent is N,N-dimethylformamide (DMF).

[0015] Furthermore, in step (1), the alkali is potassium carbonate (K2CO3).

[0016] Furthermore, in step (1), the nitrogen-containing nucleophile is selected from morpholine, N-Boc-ethylenediamine, N-Boc-piperazine, N-methylpiperazine, N-cyclopropylpiperazine, diethylamine, (2S,6R)-2,6-dimethylmorpholine, 1-(methanesulfonyl)piperazine or 1-(piperazin-1-yl)ethyl-1-one.

[0017] Furthermore, in step (1), the nucleophilic substitution reaction is carried out at a temperature of 60-80°C for 1-4 hours.

[0018] Furthermore, in step (2), the reduction system is a combination of 4-hydroxydiboron and 4,4'-bipyridine.

[0019] Furthermore, in step (2), the solvent for the reduction reaction is N,N-dimethylformamide, the reaction temperature is room temperature, and the reaction time is 10-30 minutes.

[0020] Furthermore, in step (3), the amidation condensation reaction is: in the presence of a condensing agent and a base, the intermediate obtained in step (2) is reacted with a carboxylic acid compound.

[0021] Furthermore, the condensing agent is HATU (O-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethylurea hexafluorophosphate).

[0022] Furthermore, the base is N,N-diisopropylethylamine.

[0023] Furthermore, the solvent for the amidation condensation reaction is ultra-dry dichloromethane, and the reaction time is 3-8 hours.

[0024] Furthermore, in step (3), the acylation reaction or sulfonation reaction is as follows: in the presence of a base, the intermediate obtained in step (2) is reacted with an acyl chloride or a sulfonyl chloride, wherein the base is triethylamine, the solvent for the acylation reaction or sulfonation reaction is ultra-dry dichloromethane, the reaction is carried out under ice bath conditions, and the reaction time is 10-30 minutes.

[0025] Furthermore, in step (3), the alkylation reaction is as follows: in the presence of a base, the intermediate obtained in step (2) is reacted with a haloalkane, wherein the base is anhydrous potassium carbonate, the solvent for the alkylation reaction is N,N-dimethylformamide, and the reaction time is 1-4 hours.

[0026] Furthermore, in step (3), the Buchwald-Hartwig coupling reaction is as follows: in the presence of a palladium catalyst, a ligand, and a base, the intermediate obtained in step (2) is reacted with an aryl halide, wherein the palladium catalyst is tris(dibenzylideneacetone)dipalladium, the ligand is Xphos, the base is cesium carbonate, the solvent for the Buchwald-Hartwig coupling reaction is 1,4-dioxane, the reaction temperature is 80-100℃, and the reaction time is 8-16 hours.

[0027] Furthermore, when the nitrogen-containing nucleophile in step (1) contains a Boc protecting group, or when the carboxylic acid compound in step (3) contains a Boc protecting group (tert-butyloxycarbonyl), a deprotection step is also included after the corresponding step: the intermediate or target compound containing the Boc protecting group is reacted in a 1,4-dioxane hydrochloric acid solution to remove the Boc protecting group, wherein the solvent for the deprotection step is dichloromethane, and the reaction time is 1-4 hours.

[0028] The present invention also provides a pharmaceutical composition using the above-mentioned quinazoline-containing compound as an active ingredient.

[0029] The present invention also provides the use of the above-mentioned quinazoline-containing compounds or the above-mentioned pharmaceutical compositions in the preparation of PRMT5 / EGFR antitumor inhibitor drugs.

[0030] Furthermore, the PRMT5 / EGFR antitumor inhibitor also includes a pharmaceutically acceptable carrier or excipient.

[0031] Furthermore, the dosage forms of the PRMT5 / EGFR antitumor inhibitor include injections, tablets, capsules, aerosols, suppositories, films, pellets, topical liniments, or ointments.

[0032] This invention relates to the preparation of pharmaceutical compositions by mixing quinazoline compounds with pharmaceutically acceptable carriers or excipients, and to the formulation of clinically acceptable dosage forms. Pharmaceutically acceptable excipients refer to any diluent, adjuvant, and / or carrier that can be used in the pharmaceutical field. Derivatives of this invention can be used in combination with other active ingredients, provided they do not produce other adverse effects, such as allergic reactions.

[0033] Specifically, the pharmaceutical compositions of the present invention can be formulated into several dosage forms, containing some commonly used excipients in the pharmaceutical field. The dosage forms described above can be injections, tablets, capsules, aerosols, suppositories, films, pellets, topical liniments, ointments, etc. The carriers used in the pharmaceutical compositions of the present invention are common types available in the pharmaceutical field, including: binders, lubricants, disintegrants, solubilizers, diluents, stabilizers, suspending agents, flavoring agents, preservatives, solvents, and matrices. The pharmaceutical formulations can be administered orally or via non-gastrointestinal routes (e.g., intravenously, subcutaneously, intraperitoneally, or locally). If certain drugs are unstable under gastric conditions, they can be formulated into enteric-coated tablets.

[0034] The present invention also provides the use of the above-described quinazoline-containing compounds or the above-described pharmaceutical compositions in the preparation of drugs for treating and / or preventing cancer.

[0035] Furthermore, the cancers mentioned include breast cancer, lung cancer, melanoma, pancreatic cancer, and prostate cancer.

[0036] The present invention also provides the use of the above-mentioned quinazoline-containing compounds or the above-mentioned pharmaceutical compositions in the preparation of medicaments for the treatment and / or prevention of proliferative diseases.

[0037] The clinical dosage of the quinazoline-containing compounds described above for patients can be appropriately adjusted based on the therapeutic efficacy and bioavailability of the active ingredients in vivo, their metabolism and excretion rates, and the patient's age, sex, and disease stage. However, the daily dose for adults should generally be 10-500 mg, preferably 50-300 mg. These preparations can be administered in divided doses at certain intervals, preferably one to six times, as directed by a doctor or pharmacist.

[0038] Compared with the prior art, the present invention has the following advantages and technical effects: (1) The present invention provides a quinazoline-containing compound that has a strong ability to inhibit epigenetic enzymes PRMT5 and EGFR kinases. Therefore, it can be used as an active ingredient to prepare a therapeutic drug for diseases caused by abnormal activation of epigenetic enzymes PRMT5 and EGFR kinases, and also shows potential application value in the preparation of drugs for the treatment and / or prevention of proliferative diseases and cancer.

[0039] (2) This invention provides important technical support for the development of more efficient and selective anticancer drugs. Attached Figure Description

[0040] The accompanying drawings, which form part of this invention, 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. In the drawings: Figure 1 Molecular docking prediction with EGFR protein in Example 38 (PDB: 1M17, yellow represents EGFR inhibitor, blue represents Example 38). Figure 2 The results of the time-concentration dependence experiment of the compound in Example 38; Figure 3 The results of AO staining experiments on MDA-MB-231 in each group; Figure 4 The results of wound healing experiments for each group treated with MDA-MB-231 are shown. Figure 5 The results of cloning experiments on MDA-MB-231 in each group; Figure 6 The results of the reactive oxygen species experiments on MDA-MB-231 in each group; Figure 7 The results of apoptosis experiments on MDA-MB-231 in each group; Figure 8 The body weights of mice in each group during the acute toxicity experiment on KM mice; Figure 9 The results of the in vivo toxicity test of each group on KM mice are shown. (A) shows the biochemical indicators of alanine aminotransferase (ALT) and aspartate aminotransferase (AST), (B) shows the biochemical indicators of lactate dehydrogenase (LDH), alkaline phosphatase (ALP), creatine kinase (CK) and blood urea nitrogen / creatinine (BUN / Cr), and (C) shows the biochemical indicators of serum creatinine (CREA), blood urea nitrogen (BUN), blood glucose (GLU) and AST / ALT ratio. Figure 10 HE staining results of various organs and tissues in mice to show the effects of biotoxicity on each group. Detailed Implementation

[0041] Various exemplary embodiments of the present invention will now be described in detail. This detailed description should not be considered as a limitation of the present invention, but rather as a more detailed description of certain aspects, features, and embodiments of the present invention.

[0042] It should be understood that the terminology used in this invention is merely for describing particular embodiments and is not intended to limit the invention. Furthermore, with respect to numerical ranges in this invention, it should be understood that each intermediate value between the upper and lower limits of the range is also specifically disclosed. Every smaller range between any stated value or intermediate value within a stated range, and any other stated value or intermediate value within said range, is also included in this invention. The upper and lower limits of these smaller ranges may be independently included or excluded from the range.

[0043] Unless otherwise stated, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. While only preferred methods and materials have been described herein, any methods and materials similar or equivalent to those described herein may be used in the implementation or testing of this invention. All references to this specification are incorporated by way of citation to disclose and describe methods and / or materials associated with those references. In the event of any conflict with any incorporated reference, the content of this specification shall prevail.

[0044] Various modifications and variations can be made to the specific embodiments described in this specification without departing from the scope or spirit of the invention, as will be apparent to those skilled in the art. Other embodiments derived from this specification will also be apparent to those skilled in the art. This specification and embodiments are merely exemplary.

[0045] The terms “include,” “including,” “have,” “contain,” etc., used in this article are all open-ended terms, meaning that they include but are not limited to.

[0046] All quinazoline compounds in this invention were prepared by the methods described in the synthetic routes of the examples, or by similar methods, which are well known to those skilled in the art of organic chemistry. All variable factors used in the synthetic routes are defined below.

[0047] In the embodiments of the present invention, the proton NMR spectra of the compounds were measured using a Bruker ARX-400, and the mass spectra were measured using an Agilent 1100 LC / MSD; all reagents used were analytical grade or chemically pure.

[0048] In the embodiments of this invention, room temperature or normal temperature refers to "25±2℃".

[0049] The technical solution of the present invention will be further illustrated by the following embodiments.

[0050] Example 1 N Preparation of 3-(4-morpholinoquinazolin-7-yl)-3-phenylpropionamide Step 1: Preparation of 4-(7-nitroquinazolin-4-yl)morpholine (intermediate A-1) The intermediate 4-(7-nitroquinazoline-4-yl)morpholine is synthesized from 4-chloro-7-nitroquinazoline. The specific reaction steps are as follows: 1 equivalent of 4-chloro-7-nitroquinazoline is placed in a round-bottom flask, followed by the addition of DMF, 2 equivalents of K₂CO₃, and 1.3 equivalents of morpholine. The flask is then placed in an oil bath preheated to 70°C and reacted for 2 hours. Thin-layer chromatography is used to confirm the complete reaction of the 4-chloro-7-nitroquinazoline. After cooling to room temperature, ice water is added to precipitate a pale yellow solid. The solid is filtered through a vacuum filtration flask, washed with ice water, and dried in an oven to obtain the pure intermediate 4-(7-nitroquinazoline-4-yl). The specific reaction formula is as follows: Step 2: Preparation of 4-morpholinoquinazoline-7-amine (intermediate A-2) The intermediate 4-morpholinoquinazoline-7-amine was prepared by reacting dried intermediate 4-(7-nitroquinazoline-4-yl) with 4-hydroxydiboron and 4,4'-bipyridine in DMF solvent at room temperature. The specific reaction steps are as follows: One equivalent of intermediate 4-(7-nitroquinazoline-4-yl) was placed in a round-bottom flask, and 4 equivalents of 4-hydroxydiboron and DMF were added. 0.2 equivalents of 4,4'-bipyridine were added while stirring at room temperature. The reaction was exothermic and proceeded for 15 minutes. Thin-layer chromatography was used to monitor the completeness of the reaction. After the reaction was complete, water was added to quench the reaction, followed by extraction three times with ethyl acetate. Residual water was removed by extraction with saturated brine. The ethyl acetate mixtures were combined, dried with anhydrous sodium sulfate, concentrated under reduced pressure, and then recrystallized from ethyl acetate and petroleum ether. The solution was filtered and dried to obtain a white solid, 4-morpholinoquinazoline-7-amine. The specific reaction formula is as follows: Step 3: N Preparation of 3-(4-morpholinoquinazolin-7-yl)-3-phenylpropionamide (A-3) The ammonium condensation was performed using HATU as the condensing agent. The specific reaction steps are as follows: One equivalent of 3-phenylpropionic acid was placed in a round-bottom flask, and ultra-dry dichloromethane and DIPEA (N,N-diisopropylethylamine) were added and dissolved by sonication. HATU was added for activation for 0.5 h, followed by the addition of 0.8 equivalents of 4-morpholinoquinazoline-7-amine and reaction for 5 h. Thin-layer chromatography was used to monitor the completion of the reaction. After the reaction was complete, saturated sodium bicarbonate aqueous solution was added to quench the reaction, and dichloromethane was added for extraction three times. The dichloromethane samples collected from the extraction were combined, dried with anhydrous sodium sulfate, concentrated under reduced pressure, and separated by column chromatography. The product was recrystallized from isopropanol and n-hexane to obtain a solid, which was then dried to obtain the target compound. N -(4-morpholinoquinazolin-7-yl)-3-phenylpropionamide, the specific reaction formula is as follows: The specific compound structure of A-3 is as follows: .

[0051] 1 H NMR (400 MHz, DMSO- d 6) δ 10.75 (s, 1H), 8.79 (s, 1H), 8.41 (s, 1H), 8.16 (d, J = 9.2 Hz, 1H), 7.57 (d, J = 9.1 Hz, 1H), 7.31 – 7.23 (m, 4H), 7.20 (d, J = 6.9 Hz, 1H), 4.17 (t, J = 4.6 Hz, 4H), 3.79 (d, J = 4.8 Hz, 4H), 2.95 (t, J =7.6 Hz, 2H), 2.76 (t, J = 7.8 Hz, 2H). Example 2 In Example 1, the only difference is that 3-phenylpropionic acid in step three is replaced with an equimolar amount of 3-(p-tolyl)propionic acid, resulting in the target compound. N -(4-morpholinoquinazolin-7-yl)-3-(p-tolyl)propionamide, with the following structural formula: .

[0052] 1 H NMR (400 MHz, DMSO- d 6) δ 10.74 (s, 1H), 8.78 (s, 1H), 8.42 (s, 1H), 8.16 (d, J = 9.2 Hz, 1H), 7.57 (d, J = 9.2 Hz, 1H), 7.23 – 7.02 (m, 4H), 4.17 (t, J = 4.5 Hz, 4H), 3.79 (t, J = 4.8 Hz, 4H), 2.89 (d, J = 7.5 Hz, 2H), 2.74 (d, J =7.6 Hz, 2H), 2.25 (s, 3H). Example 3 In Example 1, the only difference is that 3-phenylpropionic acid in step three is replaced with an equimolar amount of 3-(4-trifluoromethylphenyl)propionic acid, resulting in the target compound.N -(4-morpholinoquinazolin-7-yl)-3-(4-trifluoromethylphenyl)propionamide, with the following structural formula: .

[0053] 1 H NMR (400 MHz, DMSO- d 6) δ 10.79 (s, 1H), 8.79 (s, 1H), 8.40 (s, 1H), 8.16 (d, J = 9.2 Hz, 1H), 7.66 (d, J = 7.9 Hz, 2H), 7.57 (d, J = 9.3 Hz, 1H), 7.50(d, J = 7.9 Hz, 2H), 4.16 (d, J = 5.8 Hz, 4H), 3.78 (d, J = 5.6 Hz, 4H), 2.83 (d, J =7.7 Hz, 2H). Example 4 Same as Example 1, except that in step three, 3-phenylpropionic acid was replaced with an equimolar amount of 4-phenylbutyric acid to obtain the target compound. N -(4-morpholinoquinazolin-7-yl)-4-phenylbutyramide, with the following structural formula: .

[0054] 1 H NMR (400 MHz, DMSO- d 6) δ 10.37 (s, 1H), 8.58 (d, J = 2.5 Hz, 1H),8.22 (s, 1H), 7.97 (d, J = 9.3 Hz, 1H), 7.61 (d, J = 9.1 Hz, 1H), 7.43 – 7.08 (m,5H), 3.73 (dt, J = 37.5, 4.4 Hz, 8H), 2.64 (t, J = 7.7 Hz, 2H), 2.40 (t, J = 7.7Hz, 2H), 1.93 (p, J = 7.8 Hz, 2H). Example 5 Same as Example 1, except that in step three, 3-phenylpropionic acid was replaced with an equimolar amount of 4-(p-tolyl)butyric acid to obtain the target compound. N -(4-morpholinoquinazolin-7-yl)-4-(p-tolyl)butyramide, with the following structural formula: .

[0055] 1 H NMR (400 MHz, DMSO- d 6) δ 10.37 (s, 1H), 8.59 (s, 1H), 8.23 ​​(d, J =2.5 Hz, 1H), 7.97 (d, J = 8.5 Hz, 1H), 7.61 (dd, J = 9.0, 2.6 Hz, 1H), 7.09 (d, J =2.6 Hz, 4H), 3.73 (dt, J = 28.3, 3.3 Hz, 8H), 2.59 (t, J = 7.6 Hz, 2H), 2.38 (t, J = 7.6 Hz, 2H), 2.25 (s, 3H), 1.90 (p, J = 7.8 Hz, 2H). Example 6 Same as Example 1, except that in step three, 3-phenylpropionic acid was replaced with an equimolar amount of 5-phenylpentanoic acid to obtain the target compound. N- (4-morpholinoquinazolin-7-yl)-5-phenylpentanamide, with the following structural formula: .

[0056] 1 H NMR (400 MHz, DMSO- d 6) δ 10.36 (s, 1H), 8.57 (d, J = 2.1 Hz, 1H), 8.22 (d, J = 2.4 Hz, 1H), 7.95 (dd, J = 9.1, 2.4 Hz, 1H), 7.62 (dd, J = 9.1, 2.4Hz, 1H), 7.35 – 7.06 (m, 5H), 3.71 (dt, J = 42.3, 4.4 Hz, 8H), 2.59 (t, J= 6.8Hz, 2H), 2.41 (d, J = 7.2 Hz, 2H), 1.70 – 1.54 (m, J = 5.0, 3.8 Hz, 4H). Example 7 Same as Example 1, except that in step three, 3-phenylpropionic acid is replaced with an equimolar amount of 4-(4-bromophenyl)butyric acid, yielding the target compound 4-(4-bromophenyl)- N- (4-morpholinoquinazolin-7-yl)butyramide, with the following structural formula: .

[0057] 1 H NMR (400 MHz, DMSO- d 6) δ 10.35 (s, 1H), 8.57 (s, 1H), 8.22 (s, 1H), 7.94 (d, J = 8.9 Hz, 1H), 7.61 (d, J = 9.1 Hz, 1H), 7.48 – 7.13 (m, 4H), 3.76 (t, J = 4.6 Hz, 4H), 3.66 (t, J = 4.3 Hz, 4H), 2.60 (t, J = 7.6 Hz, 2H), 2.38 (t, J =7.5 Hz, 2H), 1.90 (p, J = 7.6 Hz, 2H). Example 8 Same as Example 1, except that 3-phenylpropionic acid in step three is replaced with an equimolar amount of 4-(4-chlorophenyl)butyric acid, yielding the target compound 4-(4-chlorophenyl)- N- (4-morpholinoquinazolin-7-yl)butyramide, with the following structural formula: .

[0058] 1 H NMR (400 MHz, DMSO- d 6) δ 10.35 (d, J = 2.6 Hz, 1H), 8.57 (d, J = 2.9Hz, 1H), 8.22 (d, J = 2.5 Hz, 1H), 7.94 (dd, J= 9.2, 2.9 Hz, 1H), 7.61 (dd, J =9.2, 2.5 Hz, 1H), 7.52 – 7.05 (m, 4H), 3.71 (dt, J = 41.6, 4.2 Hz, 8H), 2.62(t, J = 7.5 Hz, 2H), 2.38 (t, J = 7.4 Hz, 2H), 2.03 – 1.79 (m, 2H). Example 9 4-(4-aminophenyl)- N- The preparation of (4-morpholinoquinazoline-7-yl)butyramide is the same as in Example 1, except that 3-phenylpropionic acid in step three is replaced with an equimolar amount of 4-(4-Boc-aminophenyl)butyric acid, followed by a Boc removal reaction, as follows: Step four: The compound to be de-Boc obtained in step three was placed in a round-bottom flask and dissolved in 1 volume of ultra-dry dichloromethane. While stirring, 2 volumes of 1,4-dioxane hydrochloride were added, and the reaction was allowed to proceed for 2 hours. Thin-layer chromatography was used to monitor the completion of the reaction. After the reaction was complete, the precipitated solid was filtered, dissolved in saturated sodium bicarbonate solution and a small amount of methanol, and extracted three times with dichloromethane. The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The product was recrystallized from isopropanol and n-hexane, filtered, and dried to obtain the target compound 4-(4-aminophenyl)- N- (4-morpholinoquinazolin-7-yl)butyramide, with the following structural formula: .

[0059] 1 H NMR (400 MHz, DMSO- d 6) δ 10.35 (s, 1H), 8.57 (d, J = 2.0 Hz, 1H),8.21 (s, 1H), 7.96 (d, J = 9.1 Hz, 1H), 7.63 (d, J = 9.1 Hz, 1H), 6.86 (d, J = 7.7Hz, 2H), 6.50 (d, J = 7.7 Hz, 2H), 4.64 (d, J = 189.6 Hz, 2H), 3.77 (d, J = 4.9 Hz, 4H), 3.67 (t, J= 4.7 Hz, 4H), 2.46 (t, J = 7.6 Hz, 2H), 2.37 (t, J = 7.6 Hz, 2H), 1.84 (p, J = 7.6 Hz, 2H). Example 10 Same as Example 1, except that 3-phenylpropionic acid in step three is replaced with an equimolar amount of 4-(4-fluorophenyl)butyric acid, yielding the target compound 4-(4-fluorophenyl)- N- (4-morpholinoquinazolin-7-yl)butyramide, with the following structural formula: .

[0060] 1 H NMR (400 MHz, DMSO- d 6) δ 10.33 (s, 1H), 8.57 (d, J = 2.9 Hz, 1H), 8.21 (s, 1H), 7.96 (dd, J = 9.0, 2.9 Hz, 1H), 7.79 – 7.48 (m, 1H), 7.40 – 6.99(m, 4H), 3.73 (dt, J = 35.0, 4.3 Hz, 8H), 2.63 (t, J = 7.8 Hz, 2H), 2.44 – 2.31(m, 2H), 1.91 (p, J = 7.7 Hz, 2H). Example 11 The preparation method is the same as in Example 1, except that morpholine in step one is replaced with an equimolar amount of N-Boc-ethylenediamine, and then step four is performed according to Example 9 to obtain the target compound. N- (4-((2-aminoethyl)amino)quinazolin-7-yl)-4-(p-tolyl)butyramide, with the following structural formula: .

[0061] 1 H NMR (400 MHz, DMSO- d 6) δ 10.25 (d, J = 4.1 Hz, 1H), 8.37 (d, J = 2.8Hz, 1H), 8.23 ​​– 8.05 (m, 2H), 8.01 (d, J= 3.4 Hz, 1H), 7.70 – 7.51 (m, 1H), 7.10 (d, J = 2.7 Hz, 4H), 2.80 (s, 2H), 2.59 (t, J = 7.8 Hz, 2H), 2.42 – 2.33 (m,2H), 2.26 (d, J = 2.7 Hz, 3H), 1.90 (p, J = 8.0, 7.4 Hz, 2H). Example 12 The preparation method is the same as in Example 1, except that morpholine in step one is replaced with an equimolar amount of N-Boc-piperazine, and 3-phenylpropionic acid in step three is replaced with an equimolar amount of 4-(p-tolyl)butyric acid. Finally, step four is performed according to Example 9 to remove Boc, yielding the target compound. N- (4-(piperazin-1-yl)quinazolin-7-yl)-4-(p-tolyl)butyramide, with the following structural formula: .

[0062] 1 H NMR (400 MHz, DMSO- d 6) δ 10.57 (s, 1H), 8.52 (d, J = 3.2 Hz, 1H),8.21 (s, 1H), 7.90 (d, J = 9.2 Hz, 1H), 7.67 (d, J = 9.1 Hz, 1H), 7.09 (d, J = 3.2Hz, 4H), 3.61 (d, J = 6.0 Hz, 4H), 2.88 (t, J = 4.6 Hz, 4H), 2.59 (t, J = 7.7 Hz, 2H), 2.42 (t, J = 7.6 Hz, 2H), 2.25 (d, J = 3.3 Hz, 3H), 1.90 (t, J = 7.8 Hz, 2H). Example 13 The preparation method is the same as in Example 1, except that 3-phenylpropionic acid in step three is replaced with an equimolar amount of benzocyclobutane-7-carboxylic acid to obtain the target compound. N-(4-morpholinylquinazoline-7-yl)bicyclic [4.2.0]octyl-1(6),2,4-trien-7-carboxamide, with the following structural formula: .

[0063] 1 H NMR (400 MHz, DMSO- d 6) δ 10.86 (s, 1H), 8.66 (d, J = 3.3 Hz, 1H), 8.30 (d, J = 2.9 Hz, 1H), 8.07 (dd, J = 9.3, 3.4 Hz, 1H), 7.69 (d, J = 9.0 Hz, 1H), 7.21 (d, J = 23.9 Hz, 4H), 4.52 (t, J = 4.0 Hz, 1H), 3.87 (t, J = 4.6 Hz, 4H), 3.78 (d, J = 4.8 Hz, 4H), 3.52 – 3.40 (m, 2H). Example 14 The preparation method is the same as in Example 1, except that 3-phenylpropionic acid in step three is replaced with an equimolar amount of 3-(1-naphthyl)propionic acid to obtain the target compound. N- (4-morpholinoquinazolin-7-yl)-3-(1-naphthyl)propionamide, with the following structural formula: .

[0064] 1 H NMR (400 MHz, DMSO- d 6) δ 10.51 (s, 1H), 8.63 (d, J = 2.5 Hz, 1H), 8.28 (d, J = 2.5 Hz, 1H), 8.16 (d, J = 8.4 Hz, 1H), 8.01 (d, J = 9.2 Hz, 1H), 7.93 (d, J = 8.1 Hz, 1H), 7.79 (dt, J = 6.4, 3.2 Hz, 1H), 7.64 – 7.49 (m, 3H), 7.43(q, J= 2.6 Hz, 2H), 3.78 (s, 7H), 3.43 (t, J = 7.8 Hz, 2H), 2.84 (t, J = 7.9 Hz,2H), 1.22 (s, 1H), 0.84 (d, J = 6.9 Hz, 1H). Example 15 The preparation method is the same as in Example 1, except that 3-phenylpropionic acid in step three is replaced with an equimolar amount of 2-oxo-2-(phenylamino)acetic acid to obtain the target compound N. 1 -(4-morpholinoquinazolin-7-yl)-N 2 -Phenylacetamide, with the following structural formula: .

[0065] 1 H NMR (400 MHz, DMSO- d 6) δ 11.34 (s, 1H), 10.98 (s, 1H), 8.62 (s,1H), 8.48 (s, 1H), 8.20 – 7.95 (m, 2H), 7.89 (d, J = 8.0 Hz, 2H), 7.54 (dd, J =17.7, 7.7 Hz, 2H), 7.39 (q, J = 7.4 Hz, 2H), 7.18 (t, J = 7.4 Hz, 1H), 3.78 (q, J =5.9, 5.1 Hz, 4H), 3.73 (q, J = 4.8 Hz, 4H). Example 16 The preparation method is the same as in Example 1, except that 3-phenylpropionic acid in step three is replaced with an equimolar amount of phenylmethanesulfonic acid to obtain the target compound. N- (4-morpholinoquinazolin-7-yl)-1-phenylmethanesulfonamide, with the following structural formula: .

[0066] 1 H NMR (400 MHz, DMSO- d 6) δ 10.51 (s, 1H), 8.58 (d, J = 2.5 Hz, 1H), 7.98 (dd, J= 9.1, 2.6 Hz, 1H), 7.55 (s, 1H), 7.33 (q, J = 3.0 Hz, 4H), 7.26 (q, J = 3.2 Hz, 2H), 4.62 (d, J = 2.5 Hz, 2H), 3.78 (d, J = 4.9 Hz, 4H), 3.73 – 3.64(m, 4H). Example 17 Steps one and two are the same as in Example 1; Step 3: The key intermediate D-1 is the reaction of the acyl chloride and amine of the starting material D under alkaline conditions. The specific reaction steps are as follows: Take 1 equivalent of the starting material D and place it in a round-bottom flask. Add ultra-dry dichloromethane and 2 equivalents of triethylamine, and sonicate. Slowly add 1.2 equivalents of benzoyl chloride dropwise under ice bath conditions. After the addition is complete, rinse with a small amount of ultra-dry dichloromethane and continue the reaction for 15 minutes. Monitor the reaction progress using thin-layer chromatography. After the reaction is complete, the post-treatment is as follows: Quench the reaction with saturated sodium bicarbonate aqueous solution, extract three times with dichloromethane, combine the collected dichloromethane extracts, dry with anhydrous sodium sulfate, concentrate under reduced pressure, separate by column chromatography, and recrystallize the product to obtain intermediate D-1. The specific reaction formula is: Step 4: The key intermediate D-2 is obtained by the ester hydrolysis reaction of intermediate D-1. The specific reaction steps are as follows: 1 equivalent of intermediate D-1 is placed in a round-bottom flask, and an appropriate amount of methanol and water (2:1) is added to dissolve it. Then, 2 equivalents of lithium hydroxide are added, and the reaction is carried out under ice bath conditions for 2 hours. The reaction is detected by thin-layer chromatography. After the reaction is completed, the methanol is removed by vacuum concentration, and an appropriate amount of water is added. The pH is adjusted to between 1 and 2 with dilute hydrochloric acid, and a solid is precipitated. The solid is filtered, dried, and intermediate D-2 is obtained. The specific reaction formula is as follows: Step 5: The acid-amine condensation was performed using HATU as the condensing agent. The specific reaction steps are as follows: Take 1 equivalent of D-2 and place it in a round-bottom flask. Add ultra-dry dichloromethane and DIPEA and dissolve by sonication. Add HATU to activate for 0.5 h. Add 0.8 equivalents of A-2 and react for 5 h. Monitor the reaction completion by thin-layer chromatography. After the reaction is complete, the post-treatment is as follows: quench the reaction with saturated sodium bicarbonate aqueous solution, extract three times with dichloromethane, combine the collected dichloromethane extracts, dry with anhydrous sodium sulfate, concentrate under reduced pressure, separate by column chromatography, recrystallize the product from isopropanol and n-hexane to obtain a solid, dry, and obtain the target compound 1-benzoyl- N- (4-morpholinoquinazolin-7-yl)piperidine-4-carboxamide, structural formula is: .

[0067] ESI-MS m / z: 446.2192 [M+H] + , 1 H NMR (400 MHz, DMSO- d 6) δ 10.41 (s,1H), 8.57 (s, 1H), 8.22 (d, J = 2.1 Hz, 1H), 7.96 (d, J = 9.1 Hz, 1H), 7.65 (dd, J = 9.2, 2.1 Hz, 1H), 7.54 – 7.31 (m, 5H), 4.54 (s, 1H), 3.72 (dt, J = 42.5, 4.3Hz, 10H), 3.14 (d, J = 25.8 Hz, 1H), 2.89 (s, 1H), 2.72 (td, J = 11.4, 5.7 Hz, 1H), 1.88 (d, J = 53.3 Hz, 2H), 1.66 (d, J = 14.5 Hz, 2H). Example 18 The preparation method is the same as in Example 17, except that benzoyl chloride in step three is replaced with an equimolar amount of 4-fluorobenzoyl chloride to obtain the target compound 1-(4-fluorobenzoyl)- N- (4-morpholinoquinazolin-7-yl)piperidine-4-carboxamide, structural formula is: .

[0068] ESI-MS m / z: 464.2045 [M+H] +, 1 H NMR (400 MHz, Chloroform- d ) δ 10.41(s, 1H), 8.57 (s, 1H), 8.23 ​​(s, 1H), 7.95 (d, J = 9.0 Hz, 1H), 7.65 (d, J = 9.1Hz, 1H), 7.48 (t, J = 6.9 Hz, 2H), 7.26 (t, J = 8.7 Hz, 2H), 4.50 (s, 1H), 3.77(t, J = 4.5 Hz, 4H), 3.66 (t,J = 4.5 Hz, 4H), 3.16 – 2.79 (m, 2H), 2.72 (t, J =11.5 Hz, 1H), 1.90 (d, J = 33.5 Hz, 2H), 1.65 (d, J = 12.4 Hz, 2H). Example 19 The preparation method is the same as in Example 17, except that benzoyl chloride in step three is replaced with an equimolar amount of 4-methoxybenzoyl chloride to obtain the target compound 1-(4-methoxybenzoyl)- N- (4-morpholinoquinazolino-7-yl)piperidine-4-carboxamide, with the following structural formula: .

[0069] ESI-MS m / z: 476.23147 [M+H] +, 1 H NMR (400 MHz, DMSO- d 6) δ 10.40 (s,1H), 8.57 (s, 1H), 8.22 (s, 1H), 7.98 (d, J = 9.1 Hz, 1H), 7.65 (d, J = 8.9 Hz, 1H), 7.38 (d, J = 8.2 Hz, 2H), 6.99 (d, J = 8.2 Hz, 2H), 3.79 (d, J = 7.0 Hz, 6H), 3.68 (t, J = 4.6 Hz, 4H), 3.00 (s, 2H), 2.69 (d, J = 12.2 Hz, 1H), 1.88 (s, 2H), 1.63 (q, J = 12.2 Hz, 2H). Example 20 The preparation method is the same as in Example 17, except that benzoyl chloride in step three is replaced with an equimolar amount of 4-methylbenzoyl chloride to obtain the target compound 1-(4-methylbenzoyl)- N- (4-morpholinoquinazolino-7-yl)piperidine-4-carboxamide, with the following structural formula: .

[0070] ESI-MS m / z: 460.2354 [M+H] +, 1H NMR (400 MHz, DMSO- d 6) δ 10.40 (s,1H), 8.57 (s, 1H), 8.22 (d, J = 2.2 Hz, 1H), 7.97 (d, J = 9.1 Hz, 1H), 7.64 (dd, J = 9.1, 2.2 Hz, 1H), 7.40 – 7.15 (m, 4H), 4.50 (s, 1H), 3.72 (dt, J = 36.2, 4.4Hz, 9H), 2.98 (d, J = 65.2 Hz, 2H), 2.70 (tt, J = 11.6, 3.8 Hz, 1H), 2.33 (s,3H), 1.87 (s, 2H), 1.69 – 1.53 (m, 2H). Example 21 The preparation method is the same as in Example 17, except that benzoyl chloride in step three is replaced with an equimolar amount of 2-methylbenzoyl chloride to obtain the target compound 1-(2-methylbenzoyl)- N- (4-morpholinoquinazolino-7-yl)piperidine-4-carboxamide, with the following structural formula: .

[0071] 1 H NMR (400 MHz, DMSO- d 6) δ 10.42 (s, 1H), 8.57 (s, 1H), 8.21 (s, 1H), 7.96 (d, J = 9.1 Hz, 1H), 7.63 (d, J = 9.0 Hz, 1H), 7.44 – 7.07 (m, 4H), 4.61 (d, J = 12.9 Hz, 1H), 3.72 (dt, J = 38.1, 4.4 Hz, 9H), 3.06 (t, J = 12.7 Hz, 1H), 2.85(q, J = 25.4, 19.4 Hz, 1H), 2.70 (s, 1H), 2.23 (d, J = 24.3 Hz, 3H), 1.98 (d, J=13.0 Hz, 1H), 1.74 (d, J = 15.5 Hz, 1H), 1.69 – 1.40 (m, 2H). Example 22 The preparation method is the same as in Example 17, except that benzoyl chloride in step three is replaced with an equimolar amount of 3-trifluoromethylbenzoyl chloride to obtain the target compound. N- (4-morpholinoquinazolin-7-yl)-1-(3-trifluoromethylbenzoyl)piperidine-4-carboxamide, with the following structural formula: .

[0072] 1 H NMR (400 MHz, DMSO- d 6) δ 10.42 (s, 1H), 8.57 (s, 1H), 8.22 (s, 1H), 7.97 (d, J = 9.0 Hz, 1H), 7.91 – 7.56 (m, 5H), 4.54 (s, 1H), 3.77 (t, J = 4.6 Hz, 4H), 3.67 (t, J = 4.5 Hz, 4H), 3.59 (d, J = 16.1 Hz, 1H), 3.16 (s, 1H), 2.91 (s,1H), 2.72 (t, J = 11.5 Hz, 1H), 1.98 (s, 1H), 1.81 (s, 1H), 1.66 (s, 2H). Example 23 The preparation method is the same as in Example 21, except that piperidine-4-carboxylic acid ethyl ester (raw material D) in step three is replaced with an equimolar amount of 2-(piperazin-1-yl)acetic acid methyl ester, yielding the target compound 2-(4-(2-methylbenzoyl)piperazin-1-yl)- N- (4-morpholinoquinazolino-7-yl)acetamide, with the following structural formula: .

[0073] ESI-MS m / z: 475.2452 [M+H] +, 1 H NMR (400 MHz, DMSO- d 6) δ 10.18 (d, J =3.0 Hz, 1H), 8.51 (d, J= 2.9 Hz, 1H), 8.16 (s, 1H), 7.91 (dd, J = 9.1, 2.9 Hz, 1H), 7.66 (d, J = 9.0 Hz, 1H), 7.30 – 7.03 (m, 4H), 4.33 (t, J = 3.8 Hz, 1H),3.87 – 3.64 (m, 7H), 3.61 (t, J = 4.4 Hz, 4H), 3.20 (d, J = 3.0 Hz, 2H), 3.14 (q, J = 4.3 Hz, 2H), 2.56 (s, 2H), 2.16 (d, J = 3.0 Hz, 3H). Example 24 The preparation method is the same as in Example 23, except that the 2-methylbenzoyl chloride in step three is replaced with an equimolar amount of 4-methylbenzoyl chloride, yielding the target compound 2-(4-(4-methylbenzoyl)piperazin-1-yl)- N- (4-morpholinoquinazolino-7-yl)acetamide, with the following structural formula: .

[0074] 1 H NMR (400 MHz, DMSO- d 6) δ 10.25 (s, 1H), 8.59 (d, J = 2.5 Hz, 1H),8.23 (s, 1H), 7.98 (d, J = 9.1 Hz, 1H), 7.73 (d, J = 9.1 Hz, 1H), 7.27 (q, J = 7.8Hz, 4H), 3.78 (d, J = 5.5 Hz, 4H), 3.68 (d, J = 5.0 Hz, 5H), 3.65 (d, J = 2.3 Hz,1H), 3.28 (s, 2H), 2.59 (d, J = 18.1 Hz, 4H), 2.34 (s, 3H). Example 25 Steps one and two are the same as in Example 1; Step 3: The acid-amine condensation uses HATU as the condensing agent. The specific reaction steps are as follows: Take 1 equivalent of Boc-proline and place it in a round-bottom flask. Add ultra-dry dichloromethane and DIPEA and dissolve by sonication. Add HATU to activate for 0.5 h. Add 0.8 equivalents of A-2 and react for 5 h. Monitor the reaction completion by thin-layer chromatography. After the reaction is complete, the post-treatment is as follows: Quench the reaction with saturated sodium bicarbonate aqueous solution, extract three times with dichloromethane, combine the collected dichloromethane extracts, dry with anhydrous sodium sulfate, concentrate under reduced pressure, separate by column chromatography, recrystallize the product with isopropanol and n-hexane to obtain a solid, dry it, and obtain intermediate E-1. The specific reaction formula is: Step 4: De-Boc from E-1 to generate intermediate E-2 ( N- (4-morpholinoquinazoline-7-yl)pyrrolidine-2-carboxamide), reaction procedure: The compound to be de-Boc-treated was placed in a round-bottom flask, and 1 volume of ultra-dry dichloromethane was added for dissolution. While stirring, 2 volumes of 1,4-dioxane hydrochloride were added, and the reaction was allowed to proceed for 2 hours. Thin-layer chromatography was used to monitor the completion of the reaction. After the reaction was complete, the precipitated solid was filtered, dissolved in saturated sodium bicarbonate solution and a small amount of methanol, and extracted three times with dichloromethane. The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The product was recrystallized from isopropanol and n-hexane, filtered, and dried. The specific reaction formula is as follows: Step 5: Place 1 equivalent of intermediate E-2 into a round-bottom flask, add ultra-dry dichloromethane and 2 equivalents of triethylamine, and sonicate. Slowly add 1.2 equivalents of phenylmethanesulfonyl chloride dropwise under ice bath conditions. After the addition is complete, rinse with a small amount of ultra-dry dichloromethane and continue the reaction for approximately 15 minutes. Monitor the reaction progress using thin-layer chromatography. After the reaction is complete, the following post-treatment is performed: quench the reaction with saturated sodium bicarbonate aqueous solution, extract three times with dichloromethane, combine the collected dichloromethane extracts, dry with anhydrous sodium sulfate, concentrate under reduced pressure, separate by column chromatography, and recrystallize the product to obtain the target compound 1-(benzylsulfonyl)- N- (4-morpholinoquinazolin-7-yl)pyrrolidine-2-carboxamide, with the following structural formula: .

[0075] 1 H NMR (400 MHz, DMSO- d 6) δ 10.43 (s, 1H), 8.60 (s, 1H), 8.24 (s, 1H), 8.01 (d, J = 9.1 Hz, 1H), 7.67 (d, J = 9.1 Hz, 1H), 7.46 (d, J= 6.9 Hz, 2H), 7.38(q, J = 6.8 Hz, 3H), 4.63 – 4.41 (m, 2H), 4.41 – 4.26 (m, 1H), 3.77 (d, J = 5.2Hz, 4H), 3.73 (d, J = 5.3 Hz, 4H), 3.35 (d, J = 6.8 Hz, 2H), 2.24 (dd, J = 13.1,7.0 Hz, 1H), 2.08 – 1.74 (m, 3H). Example 26 The preparation method is the same as in Example 25, except that the phenylmethanesulfonyl chloride in step five is replaced with an equimolar amount of 3-(trifluoromethyl)benzoyl chloride to obtain the target compound. N- (4-morpholinoquinazolin-7-yl)-1-(3-trifluoromethylbenzoyl)pyrrolidine-2-carboxamide, with the following structural formula: .

[0076] 1 H NMR (400 MHz, DMSO- d 6) δ 10.62 (s, 1H), 8.59 (s, 1H), 8.24 (s, 1H), 8.02 (d, J = 9.0 Hz, 1H), 7.91 (dd, J = 19.6, 13.5 Hz, 3H), 7.74 (d, J = 7.7 Hz, 1H), 7.67 (d, J = 9.4 Hz, 1H), 4.65 (s, 1H), 3.78 (s, 4H), 3.73 – 3.65 (m, 4H), 3.58 (d, J = 17.9 Hz, 2H), 2.34 (s, 1H), 1.95 (d, J = 34.7 Hz, 3H). Example 27 The preparation method is the same as in Example 25, except that the phenylmethanesulfonyl chloride in step five is replaced with an equimolar amount of 2-(trifluoromethyl)benzoyl chloride to obtain the target compound. N- (4-morpholinoquinazolin-7-yl)-1-(2-trifluoromethylbenzoyl)pyrrolidine-2-carboxamide, with the following structural formula: .

[0077] 1 H NMR (400 MHz, DMSO- d 6) δ 10.70 (s, 1H), 8.59 (d, J = 8.5 Hz, 1H),8.28 (s, 1H), 8.01 (d, J = 9.2 Hz, 1H), 7.87 – 7.73 (m, 2H), 7.73 – 7.60 (m,2H), 7.55 (d, J = 7.8 Hz, 1H), 4.67 (dd, J = 8.6, 4.6 Hz, 1H), 3.77 (d, J = 5.5 Hz, 4H), 3.71 (d, J = 5.4 Hz, 4H), 3.23 (dt, J = 50.9, 8.2 Hz, 2H), 2.40 – 2.25 (m,1H), 2.00 (qd, J = 14.5, 13.4, 6.3 Hz, 3H), 1.83 (p, J = 6.5, 5.7 Hz, 1H). Example 28 The preparation method is the same as in Example 25, except that the phenylmethanesulfonyl chloride in step five is replaced with an equimolar amount of 3,5-ditrifluoromethylbenzoyl chloride, yielding the target compound 1-(3,5-di(trifluoromethyl)benzoyl)- N- (4-morpholinoquinazolin-7-yl)pyrrolidine-2-carboxamide, with the following structural formula: .

[0078] 1 H NMR (400 MHz, DMSO- d 6) δ 10.60 (s, 1H), 8.59 (s, 1H), 8.30 (s, 1H), 8.23 ​​(d, J = 4.4 Hz, 2H), 8.01 (d, J = 4.9 Hz, 1H), 7.92 (d, J = 15.4 Hz, 1H), 7.67(d, J = 9.1 Hz, 1H), 4.66 (t, J = 6.6 Hz, 1H), 3.78 (q, J= 6.1, 5.5 Hz, 4H), 3.70(d, J = 5.0 Hz, 4H), 3.62 (s, 2H), 2.35 (s, 1H), 2.00 (q, J = 8.7, 6.4 Hz, 3H). Example 29 The preparation method is the same as in Example 25, except that the phenylmethanesulfonyl chloride in step five is replaced with an equimolar amount of cyclohexaneformyl chloride to obtain the target compound 1-(cyclohexaneformyl)- N- (4-morpholinoquinazolin-7-yl)pyrrolidine-2-carboxamide, with the following structural formula: .

[0079] 1 H NMR (400 MHz, DMSO- d 6) δ 10.44 (s, 1H), 8.57 (s, 1H), 8.18 (s, 1H), 7.98 (d, J = 9.2 Hz, 1H), 7.64 (dd, J = 16.5, 9.1 Hz, 1H), 4.44 (t, J = 4.3 Hz, 1H), 3.78 (t, J = 4.6 Hz, 4H), 3.68 (d, J = 5.3 Hz, 5H), 3.62 – 3.54 (m, 1H), 2.16 (s, 1H), 2.00 (s, 1H), 1.91 (s, 2H), 1.78 – 1.65 (m, 4H), 1.63 (d, J =12.6 Hz, 1H), 1.53 (d, J = 10.7 Hz, 1H), 1.29 (d, J = 12.8 Hz, 4H), 1.15 (s, 1H). Example 30 The preparation method is the same as in Example 25, except that the phenylmethanesulfonyl chloride in step five is replaced with an equimolar amount of 9,10-dioxo-9,10-dihydroanthracene-2-formyl chloride, yielding the target compound 1-(9,10-dioxo-9,10-dihydroanthracene-2-formyl)- N- (4-morpholinoquinazolin-7-yl)pyrrolidine-2-carboxamide, structural formula is: .

[0080] 1 H NMR (400 MHz, DMSO- d 6) δ 10.66 (s, 1H), 8.59 (d, J = 2.6 Hz, 1H),8.34 – 8.27 (m, 2H), 8.24 (dt, J = 6.3, 2.7 Hz, 3H), 8.10 (dd, J = 7.7, 2.4 Hz, 1H), 8.02 (d, J = 8.4 Hz, 2H), 7.96 (dd, J = 5.9, 3.3 Hz, 2H), 4.69 (d, J = 6.9 Hz,1H), 3.83 – 3.76 (m, 4H), 3.70 (d, J = 5.0 Hz, 4H), 3.55 (d, J = 18.2 Hz, 2H),2.40 – 2.32 (m, 1H), 2.00 (d, J = 13.0 Hz, 3H). Example 31 The preparation method is the same as in Example 25, except that the phenylmethanesulfonyl chloride in step five is replaced with an equimolar amount of 4-methoxybenzenesulfonyl chloride, yielding the target compound 1-((4-methoxyphenyl)sulfonyl)- N- (4-morpholinoquinazolin-7-yl)pyrrolidine-2-carboxamide, with the following structural formula: .

[0081] 1 H NMR (400 MHz, DMSO- d 6) δ 10.40 (s, 1H), 8.60 (s, 1H), 8.21 (s, 1H), 8.02 (d, J = 9.1 Hz, 1H), 7.83 (d, J = 8.4 Hz, 2H), 7.69 (d, J = 9.1 Hz, 1H), 7.16(d, J = 8.4 Hz, 2H), 4.24 (t, J = 6.0 Hz, 1H), 3.85 (s, 3H), 3.79 (t, J = 4.8 Hz, 4H), 3.72 (d, J= 5.5 Hz, 4H), 3.55 – 3.43 (m, 1H), 3.22 (d, J = 8.0 Hz, 1H), 1.93 (s, 3H), 1.67 – 1.45 (m, 1H). Example 32 The preparation method is the same as in Example 25, except that the phenylmethanesulfonyl chloride in step five is replaced with an equimolar amount of 5-(dimethylamino)naphthalene-1-sulfonyl chloride, yielding the target compound 1-((5-(dimethylamino)naphthalene-1-yl)sulfonyl)- N- (4-morpholinoquinazolin-7-yl)pyrrolidine-2-carboxamide, with the following structural formula: .

[0082] 1 H NMR (400 MHz, DMSO- d 6) δ 10.37 (s, 1H), 8.59 (s, 1H), 8.44 (d, J =8.6 Hz, 1H), 8.35 (d, J = 8.6 Hz, 1H), 8.20 (d, J = 7.3 Hz, 1H), 8.06 (d, J = 2.5Hz, 1H), 7.96 (d, J = 9.1 Hz, 1H), 7.59 (ddd, J = 23.1, 15.9, 8.6 Hz, 3H), 7.23(d, J = 7.4 Hz, 1H), 4.59 – 4.46 (m, 1H), 3.84 – 3.75 (m, 4H), 3.70 (d, J = 4.8Hz, 4H), 3.55 (s, 1H), 3.44 (d, J = 7.8 Hz, 1H), 2.76 (s, 6H), 2.19 (d, J = 9.6Hz, 1H), 2.03 (q, J = 11.1, 9.0 Hz, 2H), 1.83 (s, 1H). Example 33 The preparation method is the same as in Example 25, except that step five is as follows: 1 equivalent of intermediate E-2 is placed in a round-bottom flask, ultra-dry DMF and 3 equivalents of anhydrous potassium carbonate are added, and 1.2 equivalents of (1-bromoethyl)benzene is slowly added dropwise at room temperature. After the addition is complete, the reaction continues for 2 hours, and the reaction progress is monitored by thin-layer chromatography. After the reaction is complete, the post-treatment is as follows: the reaction is quenched by adding saturated sodium bicarbonate aqueous solution, followed by extraction three times with dichloromethane. The collected dichloromethane extracts are combined, dried with anhydrous sodium sulfate, concentrated under reduced pressure, separated by column chromatography, and the product is recrystallized to obtain the target compound. N- (4-morpholinoquinazolin-7-yl)-1-(1-phenethyl)pyrrolidine-2-carboxamide, with the following structural formula: .

[0083] 1 H NMR (400 MHz, DMSO- d 6) δ 10.57 (dd, J = 41.1, 9.7 Hz, 1H), 8.58 (t, J =7.6 Hz, 1H), 8.32 – 8.18 (m, 1H), 7.99 (dt, J = 17.3, 8.9 Hz, 1H), 7.77 – 7.57(m, 1H), 7.49 – 7.21 (m, 4H), 6.99 (dd, J = 11.2, 7.1 Hz, 1H), 5.67 (tq, J =13.6, 6.4 Hz, 1H), 4.57 – 4.28 (m, 1H), 3.86 – 3.73 (m, 4H), 3.68 (d, J = 6.2Hz, 4H), 3.62 – 3.45 (m, 2H), 2.40 – 2.10 (m, 1H), 1.92 (ddt, J = 38.9, 13.5,6.8 Hz, 3H), 1.59 – 1.32 (m, 2H), 1.21 (d, J = 6.0 Hz, 2H). Example 34 The preparation method is the same as in Example 33, except that the (1-bromoethyl)benzene in step six is ​​replaced with an equimolar amount of 1-(bromomethyl)-4-toluene to obtain the target compound 1-(4-methylbenzyl)- N- (4-morpholinoquinazolin-7-yl)pyrrolidine-2-carboxamide, with the following structural formula: .

[0084] 1 H NMR (400 MHz, Methanol- d 4) δ 10.09 (s, 1H), 8.58 (s, 1H), 8.19 (s,1H), 7.97 (d, J = 9.2 Hz, 1H), 7.70 (d, J = 9.1 Hz, 1H), 7.27 (d, J = 7.6 Hz, 2H), 7.09 (d, J = 7.5 Hz, 2H), 3.78 (s, 5H), 3.68 (s, 4H), 3.57 (d, J = 12.9 Hz, 1H), 3.26 (s, 1H), 3.05 (s, 1H), 2.39 (d, J = 8.7 Hz, 1H), 2.20 (s, 3H), 1.92 – 1.70 (m, 3H). Example 35 The preparation method is the same as in Example 1, except that the 3-phenylpropionic acid in step three is replaced with an equimolar amount of 1-[(4-fluorophenyl)carbamoyl]cyclopropane-1-carboxylic acid to obtain the target compound. N- (4-Fluorophenyl)- N- (4-morpholinoquinazolin-7-yl)cyclopropane-1,1-dicarboxamide, with the structural formula: .

[0085] 1 H NMR (400 MHz, DMSO- d 6) δ 10.57 (s, 1H), 10.02 (s, 1H), 8.59 (s, 1H), 8.25 (s, 1H), 7.96 (d, J = 9.2 Hz, 1H), 7.71 (d, J = 9.2 Hz, 1H), 7.64 (t, J =7.0 Hz, 2H), 7.14 (t, J = 8.7 Hz, 2H), 3.77 (s, 4H), 3.69 (s, 4H), 1.91 (d, J =2.8 Hz, 2H), 1.49 (s, 4H), 1.03 (d, J = 6.0 Hz, 2H). Example 36 N- The preparation of (5-benzyloxypyridin-2-yl)-4-morpholinoquinazoline-7-amine follows the same steps one and two as in Example 1. The third step involves the synthesis of intermediate F-1, and the steps are as follows: Step 3: The synthesis of the key intermediate F-1 is carried out by F reacting with benzyl bromide under alkaline conditions. The specific reaction steps are as follows: One equivalent of the starting material F is placed in a round-bottom flask, dissolved in DMF, and anhydrous potassium carbonate is added. While stirring, 1.2 equivalents of benzyl bromide are added dropwise. After the addition is complete, the reaction continues for approximately 2 hours, monitored by thin-layer chromatography. After the reaction is complete, the post-treatment is as follows: the reaction is quenched by adding saturated sodium bicarbonate aqueous solution, and the mixture is extracted three times with dichloromethane. The collected dichloromethane extracts are combined, dried with anhydrous sodium sulfate, concentrated under reduced pressure, and separated by column chromatography to obtain the oily product F-1. The reaction equation is: Step 4: Place 1 equivalent of intermediate F-1 into a double-necked round-bottom flask containing 0.8 equivalents of A-2, add 0.1 equivalents of Xphos, 3 equivalents of cesium carbonate, and then add 0.05 equivalents of Pd2(dba)3. Seal the flask, evacuate it, and protect it with nitrogen. Then add 1,4-dioxane and react overnight in an oil bath at 90°C. Monitor the reaction by thin-layer chromatography. After the reaction is complete, the post-treatment is as follows: quench the reaction with saturated sodium bicarbonate aqueous solution, and extract three times with dichloromethane. Combine the dichloromethane extracts, dry with anhydrous sodium sulfate, concentrate under reduced pressure, separate by column chromatography, and recrystallize to obtain the target compound. N- (5-Benzyloxypyridin-2-yl)-4-morpholinoquinazoline-7-amine, with the following structural formula: .

[0086] 1 H NMR (400 MHz, DMSO- d 6) δ 9.54 (s, 1H), 8.52 (d, J = 2.5 Hz, 1H), 8.37(d, J = 2.5 Hz, 1H), 8.10 (d, J = 2.9 Hz, 1H), 7.87 (d, J = 9.0 Hz, 1H), 7.70 –7.24 (m, 7H), 6.97 (d, J = 8.7 Hz, 1H), 5.14 (s, 2H), 3.78 (t, J = 4.3 Hz, 4H), 3.62 (t, J = 4.4 Hz, 4H). Example 37 The preparation method is the same as in Example 36, except that the morpholine in step one is replaced with an equimolar amount of N-Boc ethylamine, and finally, Boc removal is performed (the method is the same as step four in Example 9) to obtain the target compound. N 4 -(2-aminoethyl)- N 7 -(5-benzyloxypyridin-2-yl)quinazoline-4,7-diamine, with the following structural formula: .

[0087] 1 H NMR (400 MHz, DMSO- d 6) δ 9.39 (d, J = 7.1 Hz, 1H), 8.33 (s, 1H), 8.22(d, J = 10.5 Hz, 2H), 8.09 (dd, J = 7.0, 3.8 Hz, 2H), 7.53 – 7.29 (m, 7H), 7.05 –6.87 (m, 1H), 5.13 (d, J = 2.5 Hz, 2H), 4.08 (s, 1H), 3.66 – 3.46 (m, 2H), 2.88(d, J = 6.8 Hz, 2H), 1.83 (d, J = 2.4 Hz, 1H). Example 38 The preparation method is the same as in Example 36, except that the morpholine in step one is replaced with an equimolar amount of N-Boc piperazine, and the final step involves de-Boc removal (the method is the same as step four in Example 9) to obtain the target compound. N- (5-Benzyloxypyridin-2-yl)-4-(piperazin-1-yl)quinazolin-7-amine, with the following structural formula: .

[0088] 1 H NMR (400 MHz, Chloroform- d ) δ 9.61 (s, 1H), 8.47 (s, 1H), 8.33 (s,1H), 8.09 (d, J = 3.0 Hz, 1H), 7.81 (d, J = 9.3 Hz, 1H), 7.58 – 7.27 (m, 7H), 6.98 (d, J= 8.8 Hz, 1H), 5.13 (s, 2H), 3.54 (q, J = 8.6, 6.5 Hz, 5H), 2.86 (t, J =4.6 Hz, 3H). Example 39 The preparation method is the same as in Example 36, except that the morpholine in step one is replaced with an equimolar amount of N-methylpiperazine to obtain the target compound. N- (5-Benzyloxypyridin-2-yl)-4-(4-methylpiperazin-1-yl)quinazolin-7-amine, with the following structural formula: .

[0089] 1 H NMR (400 MHz, DMSO- d 6) δ 9.53 (s, 1H), 8.49 (s, 1H), 8.35 (s, 1H), 8.10 (d, J = 3.5 Hz, 1H), 7.82 (d, J = 9.2 Hz, 1H), 7.59 – 7.28 (m, 7H), 6.97 (d, J = 8.9 Hz, 1H), 5.13 (s, 2H), 3.62 (d, J = 6.0 Hz, 4H), 2.23 (s, 3H). Example 40 The preparation method is the same as in Example 36, except that the morpholine in step one is replaced with an equimolar amount of N-cyclopropylpiperazine to obtain the target compound. N- (5-Benzyloxypyridin-2-yl)-4-(4-Cyclopropylpiperazin-1-yl)quinazolin-7-amine, with the following structural formula: .

[0090] 1 H NMR (400 MHz, DMSO- d 6) δ 9.51 (s, 1H), 8.48 (d, J = 2.5 Hz, 1H), 8.34(d, J = 2.8 Hz, 1H), 8.09 (d, J = 3.0 Hz, 1H), 7.82 (d, J = 9.3 Hz, 1H), 7.61 –7.25 (m, 7H), 6.96 (d, J= 9.2 Hz, 1H), 5.13 (s, 2H), 4.52 (dt, J = 30.9, 6.5 Hz, 4H), 3.66 (t, J = 4.6 Hz, 4H), 3.47 (q, J = 6.4 Hz, 1H), 2.44 (t, J = 4.8 Hz, 4H). Example 41 The preparation method is the same as in Example 36, except that the morpholine in step one is replaced with an equimolar amount of diethylamine, to obtain the target compound N. 7 -(5-benzyloxypyridin-2-yl)-N 4 N 4 -Diethylquinazolin-4,7-diamine, with the following structural formula: .

[0091] 1 H NMR (400 MHz, DMSO- d 6) δ 9.43 (s, 1H), 8.35 (d, J = 2.7 Hz, 1H), 8.21(d, J = 2.9 Hz, 1H), 8.08 (d, J = 3.1 Hz, 1H), 7.80 (dd, J = 9.3, 2.8 Hz, 1H), 7.62– 7.25 (m, 7H), 6.96 (dd, J = 8.9, 2.7 Hz, 1H), 5.13 (d, J = 2.7 Hz, 2H), 3.80 –3.57 (m, 4H), 1.39 – 1.23 (m, 6H). Example 42 The preparation method is the same as in Example 36, except that the morpholine in step one is replaced with an equimolar amount of (2S,6R)-2,6-dimethylmorpholine, to obtain the target compound. N- (5-Benzyloxypyridin-2-yl)-4-((2S,6R)-2,6-dimethylmorpholino)quinazolino-7-amine, with the following structural formula: .

[0092] 1 H NMR (400 MHz, DMSO- d6) δ 9.53 (s, 1H), 8.49 (s, 1H), 8.33 (s, 1H), 8.09 (d, J = 3.8 Hz, 1H), 7.86 (d, J = 9.1 Hz, 1H), 7.65 – 7.24 (m, 7H), 6.96 (d, J = 9.0 Hz, 1H), 5.14 (s, 2H), 4.09 (d, J = 12.9 Hz, 2H), 3.76 (q, J = 8.7, 7.5Hz, 2H), 2.81 (t, J = 11.7 Hz, 2H), 1.13 (d, J = 6.1 Hz, 6H). Example 43 The preparation method is the same as in Example 36, except that the morpholine in step one is replaced with an equimolar amount of 1-(methanesulfonyl)piperazine to obtain the target compound. N- (5-Benzyloxypyridin-2-yl)-4-(4-Methanesulfonylpiperazin-1-yl)quinazoline-7-amine, with the following structural formula: .

[0093] 1 H NMR (400 MHz, DMSO- d 6) δ 9.52 (d, J = 8.8 Hz, 1H), 8.68 – 8.44 (m,1H), 8.36 (d, J = 8.0 Hz, 1H), 8.25 – 7.99 (m, 1H), 7.97 – 7.76 (m, 1H), 7.68 –7.28 (m, 7H), 7.08 – 6.88 (m, 1H), 5.13 (d, J = 3.3 Hz, 2H), 3.97 – 3.61 (m,4H), 2.95 (q, J = 4.4, 3.5 Hz, 3H). Example 44 The preparation method is the same as in Example 36, except that the morpholine in step one is replaced with an equimolar amount of 1-(piperazin-1-yl)ethyl-1-one, yielding the target compound 1-(4-(7-(((5-benzyloxypyridin-2-yl)amino)quinazolin-4-yl)piperazin-1-yl)ethyl-1-one, with the structural formula: .

[0094] 1 H NMR (400 MHz, DMSO- d 6) δ 9.53 (s, 1H), 8.51 (s, 1H), 8.35 (d, J = 2.4Hz, 1H), 8.10 (d, J = 3.0 Hz, 1H), 7.88 (d, J = 9.1 Hz, 1H), 7.70 – 7.25 (m, 7H), 6.96 (d, J = 9.2 Hz, 1H), 5.14 (s, 2H), 3.83 – 3.50 (m, 8H), 2.05 (s, 3H). Performance testing I. In vitro antitumor cell activity test: (1) The quinazoline-containing compounds prepared in Examples 1-44 were tested for their in vitro inhibitory activity against breast cancer cells MCF-7, MDA-MB-231, A549, and HeLa. The reference standard was JNJ-64619178 (purchased from Shanghai Bid Pharmaceutical Technology Co., Ltd., CAS No.: 2086772-26-9, structural formula: A control group without medication was also set up (the same applies below).

[0095] The testing steps are as follows: (a) Plating: When the cell line density in the culture flask reaches 80%, digestion and centrifugation are performed. The supernatant after centrifugation is removed, and the cells at the bottom are retained. Then, 1 mL of culture medium is added to a 5 mL centrifuge tube, and the cell suspension is injected into a counting chamber after rehydration. The cells are then observed and counted under a microscope. Based on the counting results, the cell suspension concentration is adjusted to 20,000-25,000 cells / mL. Next, 180 µL of PBS is added to rows A and H and column 1 of a 96-well plate, and 180 µL of the rehydrated cell suspension is added to rows B and G. Finally, the 96-well plate is incubated at 37°C in a 5% CO2 incubator for 24 h. When the cell density reaches half capacity, the drug addition procedure can be performed.

[0096] (b) Drug Addition: There are 5 drug concentrations available. One drug is prepared at 5 concentrations. First, dissolve 1 mg of the drug in a 1.5 mL EP tube, then add 20 µL of DMSO (specifically for cell cryopreservation). Use a pipette to mix the drug thoroughly to promote dissolution. Then add 980 µL of serum-free culture medium and mix thoroughly to form a stock solution. Next, prepare the daughter solutions: Take 200 µL of the higher concentration drug solution from a 1.5 mL EP tube and add it to 400 µL of serum-free culture medium, diluting four times in total, each time by a 3-fold ratio. Add the drug sequentially from low to high concentration, adding 20 µL of each concentration to two replicate wells. The seventh well is left untreated as a control. Each 96-well plate can test four drugs. Finally, incubate the 96-well plate at 37°C and 5% CO2 for 72 hours.

[0097] (c) Preparation of MTT solution: Under light-protected conditions, add commercially available MTT powder to a 50 mL aliquot tube and add PBS to bring the volume to 50 mL. Then, filter the solution through a 0.22 µM filter membrane into a centrifuge tube, wrap it with aluminum foil, and store it in a refrigerator at 4 °C.

[0098] (d) Adding MTT solution: After 72 hours of drug treatment, remove the liquid from the 96-well plate and add 20 µL of the prepared MTT solution to each well containing cells. Then, incubate the 96-well plate in an incubator for 4 hours. After incubation, remove the MTT solution, add DMSO to each well, mix by magnetic oscillation, and then measure the OD value of each well using an enzyme-linked immunosorbent assay (ELISA) reader.

[0099] (e) The formula for calculating the inhibition rate (IR%) is: ; Table 1 shows the inhibitory activity of quinazoline-containing compounds prepared in Examples 1-44 on breast cancer cells MCF-7, MDA-MB-231, A549, and HeLa, where NA represents IC50. 50 If the value is greater than 100 μM, ND indicates that it was not detected.

[0100] Table 1. In vitro antiproliferative activity of Examples 1-44 II. Evaluation of PRMT5 enzyme inhibitory activity The inhibitory activity of all target compounds against PRMT5 enzyme was evaluated at a concentration of 10 μM using radioisotope labeling or AlphaLISA biochemical detection. JNJ-64619178 and EPZ015666 (purchased from Shanghai Bid Pharmaceutical Technology Co., Ltd., structural formula: [insert structural formula here]) were used as examples. The inhibition rates of the two positive controls were 99.5% and 98.6%, respectively, as shown in Table 2.

[0101] Table 2 Inhibition rates of Examples 1-44 Based on the preliminary screening results above, compounds with an enzyme inhibition rate exceeding 50% (Examples 20, 37, and 38) were selected for further determination of their half-maximal inhibitory concentration (IC50) against PRMT5. 50 The results are shown in Table 3.

[0102] Table 3 ICs for PRMT5 in Examples 20, 37, and 38 50 Based on the above research, it was found that Example 38 had an effect on cell viability (IC50 of MDA-MB-231). 50 =0.600μM) is greater than PRMT5 enzyme activity (IC50). 50 =6.10 μM), therefore, a kinase selectivity test was performed on Example 38. Thirty kinases were selected for testing, and it was found that Example 38 had a good inhibition rate against EGFR (78.4% inhibition rate at a concentration of 10 μM), and its activity was tested, and the activity is shown in Table 4.

[0103] Table 4 IC50 of compounds against EGFR 50 The molecular docking prediction with EGFR protein in Example 38 (PDB:1M17) is shown below. Figure 1 (Yellow represents EGFR inhibitors, blue represents Example 38) The detection of EGFR activity showed that Example 38 had good activity, reaching 0.319 μM, indicating that Example 38 has good inhibitory activity against EGFR. Molecular docking analysis showed that Example 38 filled the EGFR active pocket well, with an affinity of -9.2, and formed a one-molecule hydrogen bond with Thr766 and Glu738 in the pocket, respectively, so that the configuration was stable in the EGFR active pocket.

[0104] ADME Prediction in Examples 36-44 In recent years, network pharmacology has flourished. Through virtual ADME prediction, candidate components with excellent absorption, distribution, metabolism, and excretion properties can be efficiently screened in the early stages of drug development, rapidly narrowing down the screening range of active molecules. This allows for precise targeting of the most promising compounds. Furthermore, ADME prediction has a certain degree of accuracy and can, to some extent, replace early experimental screening, providing reliable data support for subsequent efficacy evaluation, target prediction, and mechanism of action studies. This significantly improves the efficiency and reliability of network pharmacology analysis, providing a reliable theoretical basis for subsequent mechanism of action studies and experimental validation, and significantly reducing the time and economic costs of traditional trial-and-error screening. Therefore, ADME predictions were performed on the compounds in Examples 36-44, and the ADME prediction results are shown in Table 5.

[0105] Table 5 ADME Predictions for Examples 36-44 The parameters in the table are explained as follows: Absorption (HIA%) represents the absorption by the human intestine. The higher the percentage, the better the absorption. A value greater than 80% indicates high absorption by the human body. This indicator directly reflects the predicted bioavailability of orally administered drugs, representing the ability of a compound to enter the bloodstream after oral administration. HIA% is a key parameter for evaluating the crossing of the intestinal barrier by orally administered drugs into systemic circulation, directly related to whether the drug can reach the target site at a sufficient concentration to exert its therapeutic effect. In virtual ADME screening, priority is given to highly absorbed compounds with HIA% ≥ 80%, which helps to screen candidate components with good oral bioavailability and avoid insufficient efficacy due to poor absorption. Focusing on this indicator can help screen small molecule compounds that are more suitable for oral administration at an early stage, providing a reliable material basis for subsequent efficacy validation and clinical application translation. Distribution (VDss (L / kg)) represents the ability of orally administered small molecule drugs to distribute in vivo. A value between 0.6 and 1 L / kg indicates good tissue distribution in the human body, allowing the drug to rapidly reach its target. Volume of distribution reflects the distribution of the drug between plasma and tissues; a moderate and stable distribution helps the drug achieve effective concentrations in the target organs. A VDss that is too low (<0.6 L / kg) suggests that the drug mainly remains in the blood, with weak tissue penetration, making it difficult to act on deep targets; a VDss that is too high (>1 L / kg) may cause excessive tissue accumulation, leading to off-target distribution or delayed clearance. Therefore, controlling the volume of distribution within an ideal range helps in screening candidate compounds with reasonable tissue distribution and good targeting. Metabolism refers to the seven non-inhibitory metabolic markers: CYP2D6 substrate, CYP3A4 substrate, CYP1A2 inhibitor, CYP2C19 inhibitor, CYP2C9 inhibitor, CYP2D6 inhibitor, and CYP3A4 inhibitor. The higher the inhibitor value, the more CYPs are not inhibited, indicating a lower risk of drug interactions. These seven indicators cover key members of the CYP450 enzyme family, involving the substrate state and common inhibitory effects of major metabolic enzymes (CYP3A4, CYP2D6, etc.). If a compound simultaneously meets the characteristics of being a "non-major substrate" and a "non-strong inhibitor," the risk of interfering with the metabolism of other drugs is lower when used in combination. In network pharmacology screening, prioritizing components with high metabolic safety helps improve the clinical translation potential of candidate compounds and provides a safety basis for research on compound formulations or combination therapies. Excretion (Total Clearance) represents the ability to be cleared from the body, with an ideal range of 2-10 ml / min / kg, indicating that the human body has a good ability to excrete drugs. The optimal range is 3-7 ml / min / kg, indicating that the drug has a good clearance capacity in the human body.This parameter comprehensively reflects the clearance efficiency of drugs via the liver and kidneys, and is an important basis for assessing the risk of accumulation in vivo and formulating dosing regimens. In virtual ADME screening, a clearance rate that is too low (<2 mL / min / kg) is likely to lead to drug accumulation and increase the risk of toxic side effects; while a clearance rate that is too high (>10 mL / min / kg) may result in insufficient in vivo exposure, affecting the sustained efficacy of the drug. Therefore, controlling the total clearance rate within an ideal range helps to screen candidate compounds that combine safety and efficacy.

[0106] III. Time-Concentration Dependence Experiment MDA-MB-231 cells in logarithmic growth phase were harvested and their density adjusted to 5 × 10⁻⁶. 4 100 μL / well was inoculated into a 96-well plate (5 × 10⁶ cells / mL). 3 Cells / well), with 3 replicates. After 24 h of culture, the old culture medium was discarded, and 100 μL of complete culture medium (containing 2% FBS) containing different concentrations of the compound from Example 38 (0.00123 mg / ml, 0.0037 mg / ml, 0.011 mg / ml, 0.033 mg / ml, 0.1 mg / mL) was added to each well. The drug was added to the first plate 24 h after drug addition, and the OD value was measured on the third day after drug addition, i.e., 72 h. After 24 h after drug addition to the first plate, the drug was added to the second plate, and after 48 h after drug addition to the first plate, the drug was added to the third plate. In this way, the drug had been in contact with the cells for 48 h in the second plate and 24 h in the third plate. The OD values ​​of the three plates were measured simultaneously, and after culturing for 24 h, 48 h, and 72 h respectively, 20 μL of MTT solution (5 mg / mL) was added to each well, and incubation continued for 4 h. Discard the supernatant, add 150 μL LDMSO to each well, and shake for 10 min to fully dissolve the formazan crystals. Measure the absorbance (OD) at 570 nm using a microplate reader. Calculate cell viability: Viability (%) = (OD drug group - OD blank group) / (OD control group - OD blank group) × 100%. Plot a growth inhibition curve with concentration on the x-axis and viability on the y-axis, and calculate the half-maximal inhibitory concentration (IC50) at each time point. 50 ).

[0107] Figure 2 The results of the time-concentration dependent assay of the compound in Example 38 show that the inhibitory effect of Example 38 on cells exhibits a good time-concentration dependent effect. In the high concentration range (≥0.1 mg / mL), the inhibition rate tends to saturate, reaching a maximum inhibition level of about 90%.

[0108] IV. AO staining experiment MDA-MB-231 cells in logarithmic growth phase were harvested and their density adjusted to 2 × 10⁻⁶. 5 1 mL / well was inoculated into a 24-well plate (2 × 10⁶ cells / mL).5 (Number of cells / well). After culturing for 24 h, the culture medium was discarded, and complete culture medium containing different concentrations of the compound from Example 38 (0, 1.5, 3.0, 6.0 μM) was added, with JNJ-64619178 (abbreviated as JNJ178, 3 μM) as a control, 1 mL per well, with 3 replicates for each concentration. After culturing for another 24 h, the culture medium was discarded, and the cells were washed twice with PBS. 100 μL of AO staining solution (10 μg / mL, diluted with PBS) was added to each well, and the cells were incubated at room temperature in the dark for 15 min. The staining solution was discarded, and the cells were washed three times with PBS. The cells were immediately observed and photographed under an inverted fluorescence microscope (excitation wavelength 488 nm, emission wavelength 530 nm). Normal cell nuclei showed uniform green fluorescence, while apoptotic cell chromatin was condensed or fragmented, showing bright green or orange-red fluorescence.

[0109] Figure 3 The results of the AO staining experiment on MDA-MB-231 cells in each group (where the control group is the compound concentration of 0, the same below) show that Example 38 can induce MDA-MB-231 cell apoptosis in a concentration-dependent manner, and the apoptosis morphology is more typical and the degree of apoptosis is more severe as the concentration increases.

[0110] V. Cell Migration (Scratch) Assay Draw three parallel lines on the back of the 6-well plate using a marker pen. Inoculate MDA-MB-231 cells at a rate of 5 × 10⁻⁶. 5 Cells were seeded per well. When the cells reached confluence of over 90%, a 200 μL sterile pipette tip was used to make a vertical line scratch on the back side. The cells were washed with PBS to remove suspension, and serum-free medium containing different concentrations of the compound from Example 38 (0, 0.75, 1.5, 3.0 μM) was added. JNJ-64619178 was used as a positive control at a concentration of 3 μM. The scratch width was observed and photographed under an inverted microscope at 0 h and 24 h.

[0111] Figure 4The results of wound healing experiments on MDA-MB-231 cells in each group show that after 24 h of culture, the control group cells showed significant wound healing, while the positive control drug JNJ-64619178 at a dose of 3 μM significantly inhibited wound healing. Experiments in Example 38 at doses of 0.75, 1.5, and 3 μM revealed that Example 38 inhibited the migration of MDA-MB-231 cells in a dose-dependent manner. At a concentration of 3 μM, the wound healing efficiency of Example 38 was lower than that of the positive control drug, and the inhibitory effect gradually increased with increasing concentration. In conclusion, Example 38 significantly inhibits the migration ability of MDA-MB-231 cells and exhibits superior inhibitory potential compared to the positive control at high concentrations, providing preliminary experimental evidence for its future application in anti-tumor metastasis research.

[0112] VI. Plate Cloning Experiment Logarithmically growing MDA-MB-231 cells were digested with trypsin and resuspended in complete culture medium, adjusting the density to 200 cells / mL. 2 mL of cell suspension (400 cells / well) was added to each well of a 6-well plate, gently shaken, and incubated for 24 h to allow cell adhesion. The culture medium was discarded, and complete culture medium containing different concentrations of the compound from Example 38 (0, 1.0, 2.0, 4.0 μM) was added. JNJ-64619178 was used as a positive control at a concentration of 2 μM. 2 mL was added to each well, with three replicates for each concentration. The cells were cultured for another 10 days, with the drug-containing medium changed every 3 days. When visible cell clones appeared in the wells, the culture medium was discarded, and the cells were washed twice with PBS. 1 mL of 4% paraformaldehyde was added to each well for fixation for 15 min. The fixative was discarded, and 1 mL of 0.1% crystal violet staining solution was added for staining for 30 min. Excess stain was removed by slow rinsing with running water, and the cells were dried at room temperature before photographing.

[0113] Figure 5 The cloning results for each group acting on MDA-MB-231 cells show that Example 38, at a concentration of 1 μM, effectively reduced colony formation compared to the control group, and significantly inhibited MDA-MB-231 cell colony formation in a concentration-dependent manner, inhibiting cell growth and inducing apoptosis. At a concentration of 2 μM, Example 38's ability to effectively inhibit colony formation was slightly lower than that of the positive control drug JNJ64619178. When the concentration of Example 38 was 4 μM, it significantly inhibited MDA-MB-231 cell colony formation. In conclusion, Example 38 can effectively inhibit MDA-MB-231 cell colony formation, inhibit cell growth, and induce apoptosis.

[0114] VII. Reactive Oxygen Species Experiment Take MDA-MB-231 cells in logarithmic growth phase, at 2×10⁻⁶ 5 Each probe was seeded per well in a 24-well plate. After 24 h of culture, complete culture medium containing different concentrations of Example 38 (1, 2, 4 μM) was added, with the positive group JNJ-64619178 having a concentration of 2 μM. Culture was continued for another 24 h. The culture medium was discarded, and the plates were washed twice with PBS. 500 μL of serum-free culture medium containing 10 μM DCFH-DA probe was added to each well, and the plates were incubated at 37°C in the dark for 30 min. The probe solution was discarded, and the plates were washed three times with PBS. Immediately, the plates were observed and photographed under an inverted fluorescence microscope (excitation wavelength 488 nm, emission wavelength 525 nm).

[0115] Figure 6 The results of the reactive oxygen species (ROS) experiments on MDA-MB-231 cells in each group show that JNJ-64619178 at 3 μM produced a small amount of green fluorescence, proving that the positive control drug JNJ-64619178 can promote the production of ROS in cells. After treatment with Example 38, the intensity of green fluorescence increased with increasing concentration. Example 38 at a concentration of 1.5 μM produced a large amount of ROS, which was superior to JNJ-64619178 at 3 μM. The fluorescence produced by Example 38 at 3 μM and 6 μM was more significant, showing a clear concentration gradient effect. In summary, Example 38 can significantly induce an increase in intracellular ROS levels in MDA-MB-231 cells in a concentration-dependent manner, which may be one of the important mechanisms by which it exerts its anti-tumor effect.

[0116] VIII. Apoptosis Experiment Take MDA-MB-231 cells in logarithmic growth phase, at 2×10⁻⁶ 5 Cells were seeded per well in 6-well plates. After 24 h of culture, 2 mL of complete culture medium containing different concentrations of Example 38 (2.5, 7.5, and 15 μM) was added to each well, along with a positive control group (JNJ-64619178, 2.5 and 7.5 μM). After 48 h of culture, cell culture was collected in centrifuge tubes (containing floating cells). Adherent cells were digested with trypsin without EDTA, collected, and centrifuged at 1000 rpm for 5 min, discarding the supernatant. Cells were washed twice with pre-chilled PBS, resuspended in 500 μL of binding buffer, and then 5 μL of Annexin V-FITC and 5 μL of PI were added, gently mixed, and incubated at room temperature in the dark for 15 min. Flow cytometry was used for detection within 1 h. Detection parameters were set as follows: excitation wavelength 488 nm, FITC emission wavelength 530 nm, and PI emission wavelength >600 nm. Ten cells were collected from each sample. 4 Cells were analyzed using FlowJo 10.0 software to detect early apoptosis (Annexin V).+ / PI - ) and late apoptosis (Annexin V) + / PI + The proportion of cells.

[0117] Figure 7 The results of the apoptosis experiments on MDA-MB-231 in each group show that Example 38 induced early and late apoptosis. At a concentration of 7.5 μM, early apoptotic cells Q3 accounted for 52.9%, and late apoptotic cells Q2 accounted for 39.5%. When the concentration increased to 15 μM, the proportion of cells in the Q3 region increased. In summary, this indicates that Example 38 can effectively induce early and late apoptosis, with a higher proportion of early apoptosis.

[0118] IX. Acute toxicity test in KM mice Sixteen KM mice were randomly divided into four groups (n=4): a blank control group (5% + PEG400 50% + physiological saline 45%), the low-dose group (37.5 mg / kg) of Example 38, the medium-dose group (75 mg / kg) of Example 38, and the high-dose group (150 mg / kg) of Example 38. Each group was administered the drug by gavage for 7 consecutive days, once daily. During the administration period, changes in general behavior, diet, and fur were observed, and body weight was measured daily. After the last administration, mice were fasted for 12 hours but allowed free access to water. Blood was collected by enucleation, serum was separated, and alanine aminotransferase (ALT), aspartate aminotransferase (AST), creatinine (CREA), and blood urea nitrogen (BUN) were measured using an automated biochemical analyzer. Mice were euthanized by cervical dislocation, and the heart, liver, spleen, lungs, and kidneys were dissected, weighed, and their organ indices (organ weight / body weight × 100%) were calculated. After the organ tissues were fixed with 4% paraformaldehyde for 24 hours, the tissues were sent to Nanchang Saiweier Technology Co., Ltd. for tissue sectioning and HE staining.

[0119] Figure 8 The results of the acute toxicity test on KM mice in each group show that, compared with the blank control group, there was no significant difference in the rate of weight gain of mice in each dose group of Example 38, indicating that Example 38 had no significant inhibitory effect on mouse growth in the dose range of 37.5 to 150 mg / kg.

[0120] Figure 9The results of the in vivo toxicity test in KM mice are shown in Figure 38. (A) shows the biochemical indicators of alanine aminotransferase (ALT) and aspartate aminotransferase (AST); (B) shows the biochemical indicators of lactate dehydrogenase (LDH), alkaline phosphatase (ALP), creatine kinase (CK), and blood urea nitrogen / creatinine (BUN / Cr); and (C) shows the biochemical indicators of serum creatinine (CREA), blood urea nitrogen (BUN), blood glucose (GLU), and AST / ALT ratio. It can be seen that no abnormalities were observed in any of the indicators in the positive control group (JNJ-64619178 (75 mg / kg)). The results indicate that continuous administration for 7 days in Example 38 had no significant effect on the liver and kidney function of KM mice.

[0121] 10. HE staining experiment: (1) Tissue processing: After the administration of drugs to mice in the treatment groups (37.5, 75 and 150 mg / kg) and the blank group, the organs were dissected and collected, and washed with physiological saline. The water in the mice organs after washing with physiological saline was absorbed, and then soaked in 4% formaldehyde solution for 24 h. After soaking, the organs were dehydrated by 50%, 70%, 80% and 90% alcohol for 1 h in sequence.

[0122] (2) Sample clearing and paraffin treatment: The dehydrated tissue was cleared twice in xylene solution, 15 min each time. Then the tissue was soaked in paraffin solution for 2 h.

[0123] (3) Dewaxing of paraffin sections: The paraffin sections were placed in xylene I, xylene II, anhydrous ethanol I, anhydrous ethanol II and 75% alcohol in sequence for 20, 20, 5, 5 and 5 minutes respectively, and finally rinsed with tap water.

[0124] (4) Hematoxylin staining: stain with hematoxylin solution for 3-5 min, rinse with tap water after staining, differentiate with differentiation solution, rinse with tap water again, then use blue solution to blue, and then rinse with running water.

[0125] (5) Eosin staining: The sections were dehydrated in 85% and 95% alcohol for 5 minutes each, and then stained in eosin staining solution for 5 minutes.

[0126] (6) Dehydration and mounting: The sections were placed in EtOH I-EtOH II-EtOH III-Xylene I-Xylene II for 5 min each, and then mounted with neutral resin.

[0127] (7) Observe the slides under a microscope, take pictures, save the pictures, and analyze the experimental results.

[0128] Figure 10HE staining results of various organs and tissues in mice after treatment with biotoxicity in each group show that, in the blank control group, the heart, liver, spleen, lung, and kidney tissues have clear structures and no abnormal changes. In Example 38, myocardial fibers in each dose group were neatly arranged with clear striations, and no degeneration, necrosis, or inflammatory cell infiltration was observed; liver lobule structure was intact, hepatocyte cords were normally arranged, hepatic sinusoids were not dilated, and no inflammatory infiltration was observed in the portal areas; the red and white pulp boundaries of the spleen were clear, splenic nodules were normal, and no congestion or fibrosis was observed; alveolar walls were not thickened, alveolar cavities were clear, and no inflammatory exudate or edema was observed; glomeruli were morphologically normal, renal tubular epithelial cells were not swollen or necrotic, and there was no inflammatory cell infiltration in the interstitium. No obvious pathological changes were also observed in any organ in the positive control group JNJ-64619178.

[0129] Application Example 1: Tablets 10g of the compound prepared in Example 1 was mixed with 20g of excipients according to the general pharmaceutical tableting method and then compressed into 100 tablets, each weighing 300mg.

[0130] Application Example 2: Capsules 5g of the compound prepared in Example 10 was mixed with 10g of excipients according to the requirements of pharmaceutical capsules and then filled into empty capsules, each weighing 300mg.

[0131] Application Example 3: Ointment The compound prepared in Example 12 was finely ground and then mixed with 500g of an oily matrix such as petrolatum.

[0132] Application Example 4: Aerosol Dissolve 10g of the compound prepared in Example 15 in an appropriate amount of propylene glycol, add distilled water and other excipients, and prepare a 500mL clear solution.

[0133] Application Example 5: Suppositories After grinding 10g of the compound prepared in Example 20 into a fine powder, an appropriate amount of glycerin was added and ground evenly. Then, melted glycerin gelatin was added and ground evenly. The mixture was poured into a mold coated with lubricant to obtain 50 suppositories.

[0134] Application Example 6: Droplets After heating and melting 5g of the compound prepared in Example 22 and 25g of a matrix such as gelatin, the mixture was added dropwise into low-temperature liquid paraffin to obtain 1000 pellets.

[0135] Application Example 7: Topical Liniments 10g of the compound prepared in Example 23 was mixed and ground with 2.5g of excipients such as emulsifiers according to conventional pharmaceutical methods, and then distilled water was added to 200mL to obtain the final product.

[0136] Application Example 8: Injectables Using 6g of the compound prepared in Example 26, the compound was adsorbed onto activated carbon according to conventional pharmaceutical methods, filtered through a 0.65μm microporous membrane, and then filled into a nitrogen tank to prepare an aqueous injection formulation, with each vial containing 2mL, for a total of 100 vials.

[0137] Application Example 9: Film Formulation Polyvinyl alcohol, pharmaceutical glycerin, water, etc. are stirred and expanded, then heated and dissolved. The mixture is filtered through an 80-mesh sieve. The compound prepared in Example 15 is then added to the filtrate and stirred and dissolved. 100 membranes are made by coating the membrane.

[0138] The above are merely preferred embodiments of the present invention, but the scope of protection of the present invention is not limited thereto. Any variations or substitutions that can be easily conceived by those skilled in the art within the scope of the technology disclosed in the present invention should be included within the scope of protection of the present invention.

Claims

1. A compound containing a quinazoline class, characterized in that, The structural formula is shown in Formula I: ； In Equation I, R2 is selected from , , , , , , , , , and One of them; When R2 is selected or When, R1 is selected ; When R2 is selected , , , , , , and When one of them is selected, R1 is chosen from ; When R2 is selected When, R1 is selected , , , , , , , , , , , , , , , , , and One of them; R3 is selected from , , , , and One of them; R4 is selected from and One of them; R5 is selected from , , , , , , , , and One of them.

2. A method for preparing a quinazoline-containing compound as described in claim 1, characterized in that, Starting with 4-chloro-7-nitroquinazoline, a nitrogen-containing substituent was introduced at the 4-position of the quinazoline ring via a nucleophilic substitution reaction to obtain a 4-substituted-7-nitroquinazoline intermediate; the 7-nitro group was then reduced to an amino group via a reduction reaction to obtain a 4-substituted-7-aminoquinazoline intermediate; finally, the target compound was prepared by amidation, sulfonation, alkylation, or Buchwald-Hartwig coupling reaction.

3. A pharmaceutical composition, characterized in that, The quinazoline-containing compound of claim 1 is used as the active ingredient.

4. The use of a quinazoline-containing compound as described in claim 1 or a pharmaceutical composition as described in claim 3 in the preparation of a PRMT5 / EGFR antitumor inhibitor drug.

5. The application according to claim 4, characterized in that, The PRMT5 / EGFR antitumor inhibitors also include pharmaceutically acceptable carriers or excipients.

6. The application according to claim 4, characterized in that, The dosage forms of the PRMT5 / EGFR antitumor inhibitors include injections, tablets, capsules, aerosols, suppositories, films, pellets, topical liniments, or ointments.

7. The use of a quinazoline-containing compound as described in claim 1 or a pharmaceutical composition as described in claim 3 in the preparation of a medicament for treating and / or preventing cancer.

8. The application according to claim 7, characterized in that, The cancers mentioned include breast cancer, lung cancer, melanoma, pancreatic cancer, and prostate cancer.

9. The use of a quinazoline-containing compound as described in claim 1 or a pharmaceutical composition as described in claim 3 in the preparation of a medicament for treating and / or preventing proliferative diseases.

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