Use of a quinazoline kinase inhibitor based on a chloroacetyl warhead for the preparation of a medicament for the prevention and / or treatment of a tumor and a process for its preparation

Abstract

The application discloses a use of a quinazoline kinase inhibitor based on a chloroacetyl warhead in preparation of a medicine for preventing and / or treating tumors and a preparation method thereof, and belongs to the technical field of medicines. The application provides a use of a quinazoline kinase inhibitor based on a chloroacetyl warhead in preparation of a medicine for preventing and / or treating tumors, wherein the quinazoline kinase inhibitor based on the chloroacetyl warhead is at least one selected from a compound 13, stereoisomers of the compound 13, solvates of the compound 13, pharmaceutically acceptable salts of the compound 13 or prodrugs of the compound 13, wherein the compound 13 is 4-(4-fluoro-3-chlorophenylamino)-7-(S)tetrahydrofuran-3-yloxy)-6-(N-chloroacetyl)aminoquinazoline, and the compound 13 can effectively, widely and safely inhibit tumor activity.

Description

Technical Field

[0001] This invention belongs to the field of pharmaceutical technology and relates to EGFR inhibitors, specifically to the use of a quinazoline kinase inhibitor based on a chloroacetyl warhead in the preparation of drugs for the prevention and / or treatment of tumors and its preparation method. Background Technology

[0002] For understanding the technical content of this invention: Epidermal growth factor receptor (EGFR), an important tyrosine kinase, plays a central driving role in the occurrence and development of various solid tumors, including non-small cell lung cancer (NSCLC). For EGFR mutations (such as exon 19 deletion (Del19) and L858R point mutations), small molecule tyrosine kinase inhibitors (TKIs) have become the standard clinical treatment.

[0003] Currently, three generations of EGFR-TKIs have been developed: (1) First-generation TKIs (such as gefitinib and erlotinib): These are reversible inhibitors that block EGFR activation by competitively binding to ATP. (2) Second-generation TKIs (such as afatinib and dacomitinib): These are irreversible inhibitors that incorporate an acrylamide (or similar) group as an electrophilic warhead in their molecular design. This warhead can undergo a Michael addition reaction with a conserved cysteine ​​residue (Cys797) deep within the ATP binding site of the EGFR kinase domain to form a covalent bond, thereby achieving more durable and potent inhibition. (3) Third-generation TKIs (such as osimertinib): Developed to address the T790M resistance mutation that arises after the use of second-generation drugs, these also employ an acrylamide-based Michael addition mechanism, but exhibit higher selectivity for mutant EGFR.

[0004] Among these drugs, afatinib, as a typical representative of second-generation EGFR-TKIs, has a chemical structure with quinazoline as the parent nucleus and achieves covalent binding with Cys797 through an acrylamide group. It is used clinically to treat NSCLC patients with specific EGFR-sensitive mutations.

[0005] Relevant patent documents retrieved: This document, published in China (CN 111053776 A) on April 24, 2020, discloses a pharmaceutical composition for afatinib and its application. Its purpose is to address the problems of EGFR-TKI resistance and significant toxic side effects in the treatment of non-small cell lung cancer (NSCLC), providing a pharmaceutical composition for targeted lung cancer therapy. The active ingredients of the composition are a quercetin metal complex and afatinib or a pharmaceutically acceptable salt thereof. The quercetin metal complex is selected from one of quercetin copper, quercetin manganese, and quercetin zinc. The pharmaceutical composition provided by this invention has better water solubility, lower drug resistance, higher efficacy, and fewer side effects.

[0006] Relevant non-patent literature retrieved: The article titled "The Role and Research Progress of EGFR-TKIs in Non-Small Cell Lung Cancer", Pharmaceutical Biotechnology, 2022, 29(06):634-639, published in 2022, indicates that EGFR-TKIs-targeted EGFR therapy has achieved ideal clinical efficacy. However, due to tumor heterogeneity and genomic instability, the widespread occurrence of secondary drug resistance, such as secondary gene mutations and activation of alternative signaling pathways, has greatly reduced the cure rate of non-small cell lung cancer, and the corresponding drug resistance problem continues to emerge.

[0007] The prior art represented by the aforementioned documents has at least the following unresolved technical problems or defects: Although TKIs, represented by afatinib, have achieved some clinical success, their inherent mechanisms of action and chemical structures determine that they have a series of insurmountable shortcomings and deficiencies, mainly reflected in the following aspects: (1) The inherent limitations of the mechanism of action lead to a narrow inhibitory spectrum: The covalent binding of afatinib is heavily dependent on the Michael addition reaction between the acrylamide warhead and a specific cysteine ​​residue (Cys797). The efficiency and selectivity of this reaction are highly limited by the local microenvironment of the target protein (such as steric hindrance and polarity). This dependence results in its pharmacological action being highly concentrated on EGFR family kinases, while its inhibitory activity is significantly weakened for EGFR wild-type or EGFR-independent tumor cells (such as the A549 cell line). Therefore, existing drugs have limited efficacy in treating a large number of lung cancer patients who do not carry EGFR-sensitive mutations, resulting in a narrow therapeutic spectrum and failing to meet broad clinical needs.

[0008] (2) Drug resistance is becoming increasingly serious, especially against triple mutations, which are essentially ineffective: Tumor cells evolve to develop multiple drug resistance mutations to evade drug inhibition. Among them, the EGFR T790M mutation is the most common major drug resistance mechanism after the use of first and second-generation TKIs. Although third-generation TKIs (such as osimertinib) can effectively overcome the T790M mutation, their long-term use can induce the C797S mutation, which directly eliminates the cysteine ​​residue that binds to the warhead. When tumor cells carry "triple mutations" such as Del19 / T790M / C797S, all existing TKIs that rely on Michael addition with C797 residues (including afatinib and osimertinib) will become completely ineffective. Current technologies lack solutions that can effectively address such complex multiple drug resistance mutations. Crucially, even in in vivo models, existing drugs are completely ineffective against such triple mutations. For example, in nude mouse xenograft models carrying BaF3 EGFR-Del19-T790M-C797S triple mutation cells, afatinib failed to effectively inhibit tumor growth, highlighting the extreme urgency of developing drugs with novel mechanisms of action.

[0009] (3) Insufficient inhibitory activity against EGFR-independent resistance pathways: Tumor resistance mechanisms are not limited to mutations in EGFR itself, but also include pathways independent of the EGFR signaling pathway, such as bypass activation (e.g., MET amplification) and phenotypic transformation (e.g., transformation to small cell lung cancer). Drugs such as afatinib have highly concentrated targets in the EGFR family, and their inhibitory effect on these "off-target" resistant cell lines is minimal, failing to effectively inhibit their proliferation and survival.

[0010] (4) Potential safety issues: Afatinib often causes severe adverse reactions such as diarrhea, rash, and stomatitis due to its possible off-target effects. Although its acrylamide-based reaction mechanism is intended to improve selectivity, there is still a risk of non-specific covalent binding with non-target proteins in the complex in vivo environment, which may contribute to its toxicity profile.

[0011] In summary, the core deficiency of conventional EGFR-TKIs, represented by afatinib, lies in their reliance on a covalent inhibition paradigm dependent on acrylamide-Michael addition. This paradigm exhibits inherent mechanistic deficiencies when facing EGFR wild-type tumors, complex multidrug resistance mutations (especially those involving C797S), and EGFR-independent resistance, leading to a narrow inhibitory spectrum and a lack of available drugs after resistance develops. The shortcomings of existing technologies extend beyond the cellular level to the lack of in vivo efficacy, particularly in the most challenging multidrug resistance models, where current technologies struggle to effectively inhibit tumor growth. Relevant evidence includes the afatinib drug composition described in patent publication CN 111053776 A, which primarily inhibits the A549 cancer cell line, still relying on the traditional Michael addition mechanism and failing to cover core resistant cell lines.

[0012] Therefore, there is an urgent need in this field for a covalent inhibitor with a novel mechanism of action that is not only effective at the cellular level, but more importantly, must demonstrate in vivo that it can effectively inhibit the growth of multidrug-resistant tumors caused by mutations such as C797S, and has good safety profile. Summary of the Invention

[0013] The purpose of this invention is to provide: The use of a chloroacetyl warhead-based quinazoline kinase inhibitor in the preparation of drugs for the prevention and / or treatment of tumors, its preparation method, and related technologies, in order to solve the technical problems of existing technologies, such as the inability to overcome key clinical drug resistance mutations, insufficient inhibitory activity against a wide range of EGFR types, narrow inhibitory spectrum, limited antitumor efficacy, and low safety, or combinations thereof.

[0014] Terminology Explanation: Unless otherwise defined, all technical terms in this document have the same meanings as commonly understood by one of ordinary skill in the art to which the subject matter of the claims pertains. Unless otherwise stated, all patents, patent inventions, and publications cited in this document are incorporated herein by reference in their entirety. If multiple definitions exist for terms in this document, the definitions in this chapter shall prevail.

[0015] It should be understood that the above brief description and the following detailed description are exemplary and for illustrative purposes only, and do not limit the subject matter of the invention in any way. In this invention, the singular is used in conjunction with the plural unless otherwise specifically stated. It should also be noted that, unless otherwise stated, the use of “or” or “or” means “and / or”. Furthermore, the use of the term “comprising” and other forms such as “including,” “containing,” and “contains” are not limiting.

[0016] Definitions of the standard terminology can be found in the references “Bio-targeted Therapy for Lung Cancer (2nd Edition), People’s Medical Publishing House, authors: Wu Caicun, Wu Yilong, et al., 2016-06”, “Modern Molecular Biology (5th Edition), Higher Education Press, authors: Zhu Yuxian, Li Yi, Zheng Xiaofeng, and Guo Hongwei, 2019-06-19”, and “Genetic Engineering, Higher Education Press, 2013-08-01”.

[0017] Unless otherwise stated, conventional methods within the scope of the art, such as filtration, washing, neutralization, extraction, etc., shall be used.

[0018] Unless specifically defined herein, the use of all commercially available products herein employs standard techniques. For example, it may be carried out using the manufacturer's instructions for use with the kit, or in accordance with methods known in the art or the description of this invention. The techniques and methods described herein can generally be implemented according to conventional methods well known in the art, based on the descriptions in the various summary and more specific documents cited and discussed in this specification.

[0019] The term "drug resistance" used in this article refers to the tolerance of tumor cells and other cells to the effects of drugs. Once drug resistance develops, the effectiveness of the drug decreases significantly. Drug resistance can be divided into acquired resistance and natural resistance based on its cause.

[0020] The term “potential spectrum” as used in this article refers to the range and intensity of biological activity exhibited by a drug or compound in different targets, subtypes, mutants, or disease models.

[0021] The term "stereoisomer" as used in this article refers to isomers of compounds with the same molecular formula, in which atoms or groups of atoms are connected in the same order but arranged in different spatial arrangements.

[0022] The term "solvent" as used in this article refers to a compound formed by the combination of solvent molecules and solute during the dissolution of a substance, the formation of which alters the original state of the solute.

[0023] The term "prodrug" used in this article refers to a compound that, after chemical modification, is inactive or has low activity in vitro but releases its active drug effect in vivo through enzymatic or non-enzymatic conversion.

[0024] This invention provides the use of a chloroacetyl trigger-based quinazoline kinase inhibitor in the preparation of a medicament for the prevention and / or treatment of tumors, wherein the quinazoline kinase inhibitor is selected from at least one of compound 13, a stereoisomer of compound 13, a solvate of compound 13, a pharmaceutically acceptable salt of compound 13, or a prodrug of compound 13, wherein compound 13 is 4-(4-fluoro-3-chlorophenylamino)-7-(S)tetrahydrofuran-3-yloxy)-6-(N-chloroacetyl)aminoquinazoline.

[0025] Specifically, the structural formula of compound 13 is shown in formula (I): (I).

[0026] The inventors discovered that by fundamentally modifying the substituent at the 6-position of the quinazoline nucleus, replacing the acrylamide group (Michael addition receptor) in existing technologies (represented by afatinib) with a chloroacetamido group (nucleophilic substitution reaction receptor), they could fundamentally alter the mechanism of action of quinazoline kinase inhibitors on tumors. The inventors found that compound 13 no longer relies on the Michael addition reaction with the C797 residue of EGFR kinase, but instead undergoes a nucleophilic substitution reaction (SN2) with a conserved cysteine ​​residue in the ATP binding cavity of the chloroacetyl group via the chlorine atom Cl (an excellent leaving group), forming a covalent bond. Furthermore, this novel mechanism of action may extend its inhibitory spectrum to other key kinase targets beyond EGFR, demonstrating superior inhibitory activity against EGFR wild-type cancer cells and overcoming the limitations of existing drug target populations. Simultaneously, it also exhibits superior inhibitory effects compared to existing technologies against cancer cells carrying common EGFR-sensitive mutations (such as PC-9 and Del19) and classic drug-resistant mutations (such as NCI-H1975 and L858R / T790M). Furthermore, compound 13 effectively inhibits multiple key signaling pathways driving tumor cell survival (pro-apoptosis effect) and motility (anti-migration effect). This multi-pronged approach makes its anti-tumor effect more comprehensive and thorough than afatinib, which primarily targets the EGFR family. Moreover, compound 13 has been demonstrated to possess excellent in vivo safety profiles and a broad therapeutic window.

[0027] According to some embodiments of the present invention, the tumor is selected from at least one of breast cancer, lung cancer, kidney cancer, prostate cancer, colorectal cancer, and pancreatic cancer.

[0028] Abnormal activation of EGFR, especially persistent activation of EGFR due to gene mutations, is closely related to the occurrence and development of various epithelial-derived malignancies (Research Progress on the Role of EGFR in Tumor Development and Treatment, Clinical Medical Progress, 2024, 14(12), 796-803), such as in renal cell carcinoma (Research Progress on EGFR in Renal Cell Carcinoma, Journal of Clinical Urology, 2012, 27(11):873-876), lung cancer (Research Progress on EGFR-related Inhibitors for Lung Cancer Driver Genes, Inner Mongolia Medical Journal, 2018, 50(08):908-911), prostate cancer (Research on Novel EGFR Tyrosine Kinase Inhibitors for the Treatment of Prostate Cancer, Kunming Medical University, 2019), and pancreatic cancer (Research on EGFR-ERK in Pancreatic Cancer). EGFR is overexpressed in various cancers, including breast cancer ("The Role of EGFR in Triple-Negative Breast Cancer and Research Progress in Targeted Therapy", Oncology, 2023, 43(10):829-838) and colorectal cancer ("Research Progress on Anti-EGFR Therapy Resistance Mechanism in Metastatic Colorectal Cancer", Chinese Journal of General Surgery, 2024, 33(06):996-1011). EGFR inhibitors can be used in the preparation of drugs to prevent or treat the above-mentioned EGFR-mediated diseases.

[0029] Furthermore, lung cancer can include non-small cell lung cancer.

[0030] According to some embodiments of the present invention, the tumor may be mediated by EGFR.

[0031] Furthermore, the non-small cell lung cancer includes tumors resistant to at least one of the first-generation EGFR tyrosine kinase inhibitors, the second-generation EGFR tyrosine kinase inhibitors, and the third-generation EGFR tyrosine kinase inhibitors.

[0032] Specifically, the first-generation EGFR tyrosine kinase inhibitor is selected from at least one of gefitinib, erlotinib, and icotinib.

[0033] Specifically, the second-generation EGFR tyrosine kinase inhibitor is selected from at least one of afatinib and dacomitinib.

[0034] Specifically, the third-generation EGFR tyrosine kinase inhibitor is selected from at least one of osimertinib, ametinib, and vormetinib.

[0035] For example, the EGFR mutation type in the EGFR-mutant non-small cell lung cancer is selected from at least one of T790M, C797S, L858R and 19del.

[0036] EGFR mutations include single mutations of T790M, C797S, L858R and 19del, double mutations of T790M / C797S, or triple mutations of Del19 / T790M / C797S.

[0037] Furthermore, EGFR mutations include individual mutations such as T790M, C797S, L858R, and 19del.

[0038] Furthermore, EGFR mutations include the 19Del / T790M double mutation, the L858R / T790M double mutation, the T790M / C797S double mutation, the 19Del / C797S double mutation, and the L858R / C797S double mutation.

[0039] Preferably, EGFR mutations include single mutations of T790M, C797S, L858R and 19del, double mutations of T790M / C797S, or triple mutations of Del19 / T790M / C797S.

[0040] According to some embodiments of the present invention, the non-small cell lung cancer may also be EGFR wild-type non-small cell lung cancer.

[0041] According to some embodiments of the present invention, the medicament contains a therapeutically effective amount of the quinazoline kinase inhibitor.

[0042] Further, based on the weight of the recipient, the effective therapeutic dose can be 0.1 mg / kg / day to 10 mg / kg / day. For example, the effective therapeutic dose can be 0.1 mg / kg / day, 1 mg / kg / day, 2 mg / kg / day, 3 mg / kg / day, 4 mg / kg / day, 5 mg / kg / day, 6 mg / kg / day, 7 mg / kg / day, 8 mg / kg / day, 9 mg / kg / day, 10 mg / kg / day, 12 mg / kg / day, 15 mg / kg / day, 18 mg / kg / day, 20 mg / kg / day, 22 mg / kg / day, 25 mg / kg / day, or any value within the range of any two of the above values.

[0043] Specifically, the drug also includes pharmaceutically acceptable carriers and / or excipients.

[0044] Furthermore, the carriers include, but are not limited to, nanoparticles, liposomes, polymers, micelles, cyclodextrins, exosomes, etc.

[0045] Furthermore, the drug also includes pharmaceutically acceptable excipients. Even further, the pharmaceutically acceptable excipients include one or more of the following: excipients, diluents, lubricants, antibacterial agents, suspending agents, suspending aids, wetting agents, emulsifiers, preservatives, antioxidants, buffers, solubilizers, thickeners, stabilizers, and sweeteners.

[0046] Furthermore, the dosage form of the drug is tablets, capsules, injections, powder for injection, or inhalation.

[0047] This invention also provides: a method for preparing a chloroacetyl-based quinazoline kinase inhibitor, wherein the chloroacetyl-based quinazoline kinase inhibitor is selected from at least one of compound 13, a stereoisomer of compound 13, a solvate of compound 13, a pharmaceutically acceptable salt of compound 13, or a prodrug of compound 13, wherein the method for preparing compound 13 includes the following steps: S1. 4-Chloro-6-nitro-7-fluoroquinazoline and 4-fluoro-3-chloroaniline are reacted in isopropanol with triethylamine and reacted at 50℃~70℃ for 5~7 hours to obtain 4-(4-fluoro-3-chlorophenylamino)-7-fluoro-6-nitroquinazoline; S2. The 4-(4-fluoro-3-chlorophenylamino)-7-fluoro-6-nitroquinazoline is reacted with S-(+)-3-hydroxytetrahydrofuran in a sodium hydride / tetrahydrofuran system at 90℃~110℃ for 7~9 hours to obtain 4-(4-fluoro-3-chlorophenylamino)-7-((S)tetrahydrofuran-3-yloxy)-6-nitroquinazoline; S3. The 4-(4-fluoro-3-chlorophenylamino)-7-((S)tetrahydrofuran-3-yloxy)-6-nitroquinazoline, reduced iron powder, and ammonium chloride are reacted in an ethanol-water mixed solvent at 80℃~100℃ for 5~7 hours to carry out a nitro reduction reaction, yielding 4-(4-fluoro-3-chlorophenylamino)-7-((S)tetrahydrofuran-3-yloxy)-6-aminoquinazoline; S4. Under ice bath conditions, a tetrahydrofuran solution of chloroacetyl chloride was added dropwise to the 4-(4-fluoro-3-chlorophenylamino)-7-((S)tetrahydrofuran-3-yloxy)-6-aminoquinazoline. The reaction was carried out at room temperature for 3 to 5 hours, and the mixture was purified by chromatography to obtain compound 13, which is 4-(4-fluoro-3-chlorophenylamino)-7-(S)tetrahydrofuran-3-yloxy)-6-(N-chloroacetyl)aminoquinazoline.

[0048] Specifically, the molar ratio of 4-chloro-6-nitro-7-fluoroquinazoline to 4-fluoro-3-chloroaniline in step S1 is 1:1.0 to 1.2. For example, this molar ratio can be 1:1.0, 1:1.1, 1:1.2, or any value within the range of any two of the above values.

[0049] Specifically, in step S2, the molar ratio of 4-(4-fluoro-3-chlorophenylamino)-7-fluoro-6-nitroquinazoline to S-(+)-3-hydroxytetrahydrofuran is 1:1.1 to 1.2. Exemplarily, this molar ratio can be 1:1.1, 1:1.15, 1:1.2, or any value within the range of any two of the above values.

[0050] In step S4, the molar ratio of 4-(4-fluoro-3-chlorophenylamino)-7-((S)tetrahydrofuran-3-yloxy)-6-aminoquinazoline to chloroacetyl chloride is 1:1.0 to 1.5. Exemplarily, this molar ratio can be 1:1.0, 1:1.1, 1:1.2, 1:1.3, 1:1.4, 1:1.5, or any value within the range of any two of the above values.

[0051] Examples 1 and Effect Examples 1-6 in this invention at least support the scope of protection for "the use of quinazoline kinase inhibitors in the preparation of medicaments for the prevention and / or treatment of tumors".

[0052] The term "use of quinazoline kinase inhibitors in the preparation of medicaments for the prevention and / or treatment of tumors" is derived from the foregoing explanation and / or the preparation and in vivo / in vitro experimental results of the corresponding compound 13 in Example 1 and Effect Examples 1-6. Therefore, those skilled in the art can reasonably presume that "use of quinazoline kinase inhibitors in the preparation of medicaments for the prevention and / or treatment of tumors," its subordinate concepts, its substantially equivalent technical means, and technical means that can replace it within the scope of conventional technical means and common knowledge based on the existing level of technology should all fall within the protection scope of "use of quinazoline kinase inhibitors in the preparation of medicaments for the prevention and / or treatment of tumors."

[0053] Example 1 of this invention at least supports the protection scope of "preparation method of quinazoline kinase inhibitor based on chloroacetyl warhead".

[0054] The "Preparation Method of Quinazoline Kinase Inhibitor Based on Chloroacetyl Warhead" is derived from the foregoing explanation and / or the preparation of the corresponding compound 24 in Example 1. Therefore, those skilled in the art can reasonably presume that the "Preparation Method of Quinazoline Kinase Inhibitor Based on Chloroacetyl Warhead," its subordinate concepts, its substantially equivalent technical means, and technical means that can replace it within the scope of conventional technical means and common knowledge based on the existing level of technology should all fall within the protection scope of the "Preparation Method of Quinazoline Kinase Inhibitor Based on Chloroacetyl Warhead."

[0055] The present invention has at least the following beneficial effects: Compared with the prior art, the present invention provides the use of a quinazoline kinase inhibitor based on a chloroacetyl warhead in the preparation of drugs for the prevention and / or treatment of tumors, and its preparation method, which has better technical effects, specifically reflected in the following aspects: (1) The chloroacetyl kinase inhibitor of the present invention uses a chloroacetylamino covalent warhead to act on the C797 residue through a nucleophilic substitution reaction (SN2) mechanism. At the same time, it interacts with the hydroxyl group formed by the C797S mutation by forming hydrogen bonds, thereby successfully bypassing the C797S drug resistance bottleneck. It effectively overcomes the C797S drug resistance mutation in both in vivo and in vitro models, and has a breakthrough anti-drug resistance effect.

[0056] (2) Based on the novel mechanism of action of the quinazoline kinase inhibitor described above, this invention discovers that the chlorine atom in the quinazoline kinase inhibitor, as a superior leaving group compared to the acrylamide double bond system, enables the chloroacetyl warhead to undergo a more extensive and controllable nucleophilic substitution reaction with a wider range of conserved cysteine ​​residues within the ATP binding cavity of the kinase. This covalent inhibition mechanism based on nucleophilic substitution is less dependent on the target microenvironment, thus inhibiting a wider range of kinase targets. Consequently, it also produces a strong inhibitory effect on tumor cells that do not solely rely on the classical EGFR signaling pathway (such as A549), overcoming the narrow inhibitory spectrum of existing drugs. Simultaneously, the migration process of tumor cells is synergistically regulated by multiple signaling pathways. Compound 13, by inhibiting a wider range of kinase targets, may simultaneously block multiple signaling pathways driving cell migration (such as c-Met, Axl, etc.), thereby producing a stronger anti-migration phenotype.

[0057] (3) The present invention also found that the chloroacetyl warhead of the quinazoline kinase inhibitor has higher reactivity and better pharmacokinetic properties, and its covalent bond formation with the target kinase is more efficient and faster. This more efficient and stable covalent binding leads to more thorough and longer-lasting target inhibition, thereby exhibiting a stronger anti-proliferation and pro-apoptotic effect at the cellular functional level.

[0058] (4) This invention has found that the chloroacetyl warhead of this quinazoline kinase inhibitor has a shorter alkyl chain length and higher reactivity selectivity, which may reduce the risk of nonspecific covalent binding to irrelevant proteins during in vivo circulation. Its highly effective nature allows it to take effect at lower therapeutic doses, which helps to create a wider safety window.

[0059] In summary, this invention, by introducing a chloroacetamino group at the 6-position of the quinazoline core in a quinazoline kinase inhibitor, has shown that its application in the preparation of drugs for the prevention and / or treatment of tumors can achieve a revolutionary mechanism of action and a comprehensive improvement in in vitro efficacy (overcoming ultimate drug resistance, expanding to wild-type, broad-spectrum potency, and multiple mechanisms). Ultimately, it exhibits excellent efficacy and safety in both disease models and in vivo models in healthy animals. Furthermore, these effects are interconnected, collectively giving its application in the preparation of drugs for the prevention and / or treatment of tumors greater technical effectiveness and promising prospects for technology transfer. Attached Figure Description

[0060] Figure 1 For CCK-8 assay results (A549 and NCI-H1975 cell lines), there was no significant difference in ns; **** P ≤0.0001.

[0061] Figure 2For CCK-8 assay results (PC9, EGFR Del19 - T790M-C797S / BaF3 cell lines), there was no significant difference in ns;** P ≤ 0.01; **** P ≤ 0.0001.

[0062] Figure 3 The results of the plate colony formation experiment (A549 cell line) showed no significant difference in ns; **** P ≤ 0.0001.

[0063] Figure 4 The results of the plate colony formation experiment (PC9 cell line) showed no significant difference in ns;** P ≤ 0.01; **** P ≤0.0001.

[0064] Figure 5 The results of the plate colony formation experiment (NCI-H1975 cell line) showed no significant difference in ns;* P ≤0.05;** P ≤ 0.01.

[0065] Figure 6 The results of apoptosis were detected by flow cytometry (EGFR Del19-T790M-C797S / BaF3 cell line - Control group, afatinib group and compound 13 group).

[0066] Figure 7 A bar chart showing the apoptosis rate of EGFR Del19-T790M-C797S / BaF3 cells; no significant difference was observed in ns. P ≤ 0.01; **** P ≤ 0.0001.

[0067] Figure 8 The scratch assay results (A549 cell migration inhibition) showed no significant difference in ns; **** P ≤ 0.0001.

[0068] Figure 9 The scratch assay results (NCI-H1975 cell migration inhibition) showed no significant difference in ns; *** P ≤0.001.

[0069] Figure 10 The scratch assay results (PC9 cell migration inhibition) showed no significant difference in ns;* P ≤ 0.05; *** P ≤0.001.

[0070] Figure 11 This is the curve showing the change in mouse body weight.

[0071] Figure 12 The results are the biochemical indicators of liver and kidney function in mice.

[0072] Figure 13 HE-stained sections of mouse liver and kidney tissue.

[0073] Figure 14 This is a diagram illustrating the anti-tumor effect in a BaF3 xenograft model. P ≤ 0.01; *** P ≤ 0.001; **** P ≤ 0.0001. Detailed Implementation

[0074] Unless otherwise specified, all raw materials and reagents used in this invention were purchased from commercial suppliers, and experiments were conducted in accordance with the operating instructions. Unless otherwise specified, all instruments, equipment, and apparatus used in this invention are conventional instruments, equipment, and apparatus, and experiments were conducted in accordance with the operating instructions and the accompanying reagents.

[0075] To make the objectives, technical solutions, and advantages of this invention clearer, the invention will be further described in detail below with reference to embodiments. Unless otherwise specified in the embodiments, conditions are performed under conventional conditions or conditions recommended by the manufacturer. All reagents or instruments without specified manufacturers are commercially available conventional products. Numerous specific details are provided in the following detailed embodiments to better illustrate the invention. The specific embodiments described herein are for illustrative purposes only and are not intended to constitute any limitation on the invention.

[0076] Data analysis and statistical analysis were performed using professional data processing software, and significance analysis was conducted using one-way ANOVA. P ≤ 0.05 indicates a significant difference.

[0077] The main reagents and instruments used in the following examples were: lung cancer cells A549 (Shanghai Jinyuan Biotechnology Co., Ltd., JY135), PC-9 (also known as PC9, Shanghai Jinyuan Biotechnology Co., Ltd., JY123), NCI-H1975 (also known as 1975, Shanghai Jinyuan Biotechnology Co., Ltd., JY102) and EGFR-Del19-T790M-C797S / BAF3 (also known as BaF3, Nanjing Kebai Biotechnology Co., Ltd., CBP73173).

[0078] Example 1: Quinazoline kinase inhibitor and its preparation method A quinazoline kinase inhibitor based on a chloroacetyl warhead: 4-(4-fluoro-3-chlorophenylamino)-7-(S)tetrahydrofuran-3-yloxy)-6-(N-chloroacetyl)aminoquinazoline, named compound 13, has the structural formula shown in formula (I): (I).

[0079] The basic reaction process for the preparation of compound 13 is as follows:

[0080] The specific preparation method of compound 13 includes the following steps: (1) Preparation of 4-(4-fluoro-3-chlorophenylamino)-7-fluoro-6-nitroquinazoline: 2.3 g of 4-chloro-6-nitro-7-fluoroquinazoline and 100 mL of isopropanol were added to a 250 mL round-bottom flask. After stirring evenly, 5 mL of triethylamine and 1.6 g of 4-fluoro-3-chloroaniline were added. The mixture was reacted at 60 °C for 6 hours. After the reaction was completed, the mixture was cooled and filtered to obtain a filter cake. The filter cake was washed with a small amount of methanol aqueous solution to obtain 3.0 g of the product 4-(4-fluoro-3-chlorophenylamino)-7-fluoro-6-nitroquinazoline. MS (m / z): 337.0 (M+1)+.

[0081] (2) Preparation of 4-(4-fluoro-3-chlorophenylamino)-7-((S)tetrahydrofuran-3-yloxy)-6-nitroquinazoline: 1 g of S-(+)-3-hydroxytetrahydrofuran, 500 mg of sodium hydride, and 100 mL of tetrahydrofuran were added to a 250 mL round-bottom flask. After stirring evenly, 3.3 g of 4-(4-fluoro-3-chlorophenylamino)-7-fluoro-6-nitroquinazoline was added. The mixture was reacted at 100 °C for 8 hours. After the reaction was complete, the reaction solution was poured into cold water, neutralized with hydrochloric acid solution, and filtered to obtain a filter cake. The filter cake was washed with a small amount of methanol solution to obtain 3 g of the product 4-(4-fluoro-3-chlorophenylamino)-7-((S)tetrahydrofuran-3-yloxy)-6-nitroquinazoline. MS (m / z): 405.1 (M+1)+.

[0082] (3) Preparation of 4-(4-fluoro-3-chlorophenylamino)-7-(S)tetrahydrofuran-3-yloxy)-6-aminoquinazoline: Add 4 g of 4-(4-fluoro-3-chlorophenylamino)-7-((S)tetrahydrofuran-3-yloxy)-6-nitroquinazoline, 150 mL of ethanol, 10 mL of water, 3 g of reduced iron powder, and 2 g of ammonium chloride to a 250 mL round-bottom flask. After stirring evenly, heat slowly at 90 °C for 6 hours. After the reaction is complete, filter while hot to remove excess iron powder, evaporate the solvent under reduced pressure, add 100 mL of distilled water, extract with dichloromethane, dry with anhydrous sodium sulfate, and recover the solvent under reduced pressure to dryness to obtain product 4-(4-fluoro-3-chlorophenylamino)-7-((S)tetrahydrofuran-3-yloxy)-6-aminoquinazoline 3 g. gram, MS (m / z): 375.1 (M+1)+.

[0083] (4) Preparation of 4-(4-fluoro-3-chlorophenylamino)-7-(S)tetrahydrofuran-3-yloxy)-6-(N-chloroacetyl)aminoquinazoline: Add 3.7 g of 4-(4-fluoro-3-chlorophenylamino)-7-(S)tetrahydrofuran-3-yloxy)-6-aminoquinazoline, 120 mL of tetrahydrofuran, and 5 mL of triethylamine to a 250 mL round-bottom flask. Place the flask in an ice bath, stir well, and then slowly add 30 mL of a THF solution containing 1.5 g of chloroacetyl chloride dropwise using a funnel. The addition should be completed over 30 minutes. Continue the reaction at room temperature for 4 hours. After the reaction is complete, pour the reaction solution into cold water and adjust the pH with sodium hydroxide solution until a large amount of solid precipitates. Filter to obtain the crude product, and further purify by column chromatography to obtain the product 4-(4-fluoro-3-chlorophenylamino)-7-(S)tetrahydrofuran-3-yloxy)-6-(N-chloroacetyl)aminoquinazoline. -(S)tetrahydrofuran-3-yloxy)-6-(N-chloroacetyl)aminoquinazoline 3 g, MS (m / z): 451.1 (M+1)+.

[0084] The quinazoline kinase inhibitors of this invention include, but are not limited to, at least one of compound 13, a pharmaceutically acceptable salt of compound 13, a stereoisomer of compound 13, a solvate of compound 13, or a prodrug of compound 13.

[0085] Comparative Example 1 Afatinib is provided, purchased from McLean, catalog number A877060.

[0086] Example 1: CCK-8 Experiment Lung cancer cells in logarithmic growth phase, including A549, PC-9, NCI-H1975, and BaF3 (EGFR Del19-T790M-C797S / BaF3), were washed three times with sterile 1×PBS. An appropriate amount of 0.25% trypsin was added to digest the cells. When the intercellular spaces widened and the cells became rounded and shrunken, an equal volume of complete cell culture medium containing 10% fetal bovine serum was added to terminate the digestion. The cells were centrifuged at 300 g for 5 minutes and seeded evenly in each well of a 96-well plate at a count of 5000 cells / well. After cell adhesion, the culture medium was discarded. Add 200 μL of the prepared drug solution of the corresponding concentration (compound 13 or afatinib in Comparative Example 1) to each well (the drug solution concentrations used for A549, PC-9 and NCI-H1975 cells are 0, 250 nM and 500 nM, respectively; the drug concentrations used for BaF3 cells are 0, 20 nM and 60 nM, respectively). After treating the cells with each drug for 72 hours, remove the 96-well plates of each group of cells from the cell culture incubator, add CCK8 working solution to each well, incubate for 1 hour, and measure and count the cell absorbance at a wavelength of 450 nm using a full-spectrum microplate reader.

[0087] The results showed that compound 13 exhibited significantly better inhibitory activity than the control (afatinib) in all four cell models (as shown in Figures 1-2).

[0088] Example 2: Plate colony formation experiment Three human lung cancer cell lines (A549, PC-9, and NCI-H1975) in logarithmic growth phase were digested with trypsin and prepared into single-cell suspensions. The cell suspensions were seeded at a density of 500 cells / well in 6-well plates, with 2 mL of cell suspension added to each well. The plates were gently shaken to ensure even distribution of cells at the bottom, and then incubated at 37°C in a 5% CO2 incubator until cell attachment. After cell attachment, the culture medium was discarded. 2 mL of the corresponding concentration of drug (compound 13 or afatinib from Comparative Example 1) was added to the appropriate well (0, 250 nM, and 500 nM drug solutions were prepared, respectively). After 72 hours of drug treatment, the medium was replaced with normal culture medium, and subsequently, the medium was changed every 5 days until visible cell colonies formed. After cell clones have formed, discard the culture supernatant in the 6-well plates. Gently wash the plates three times with PBS. Add 500 μL of 4% paraformaldehyde to each well and fix at room temperature for 15 minutes. After fixation, add 500 μL of crystal violet staining solution to each well and stain at room temperature for 15 minutes to ensure thorough staining of the cell clones. After staining, wash the plates repeatedly with PBS until the background is clean. Invert the plates to dry and then photograph the results.

[0089] The results showed that, compared with the control group drug (afatinib), the number of cell clones of compound 13 decreased significantly with increasing drug concentration, indicating that the inhibitory ability of compound 13 to inhibit proliferation was better than that of afatinib (as shown in Figures 3-5).

[0090] Example 3: Flow cytometry detection of apoptosis Logarithmic growth phase BaF3 cells were seeded in 6-well plates and cultured overnight. The supernatant was discarded, and the cells were washed once with PBS. The cells were then divided into a control group (complete culture medium without drug), an afatinib group, and a compound 13 group. 2 mL of the corresponding drug solution (20 nM and 60 nM) was added to each well. After co-incubation with the drug for 13 hours, the cell culture suspension was aspirated into centrifuge tubes, centrifuged at 1000 g for 5 minutes, the supernatant was discarded, and the cells were collected. The cells were gently resuspended in 195 μL of Annexin V-FITC binding solution. Then, 2.5 μL of Annexin V-FITC and 5 μL of propidium iodide staining solution were added and gently mixed. The cells were incubated at room temperature in the dark for 10-20 minutes, then placed on ice and protected from light with aluminum foil. The apoptosis rate of each group was then measured and statistically analyzed using flow cytometry.

[0091] The results showed that in BaF3 cells, compound 13 induced apoptosis significantly more strongly than the control group (afatinib). This indicates that compound 13 has a higher pro-apoptotic capacity (as shown in Figures 6-7).

[0092] Example 4: Scratch Test Collect cells in the logarithmic growth phase (A549, PC-9, NCI-H1975), wash three times with sterile 1×PBS, add an appropriate amount of 0.25% trypsin for digestion, and when the intercellular spaces widen and the cells become rounded and wrinkled, add an equal volume of complete cell culture medium containing 10% fetal bovine serum to terminate the digestion; collect the cells in sterile centrifuge tubes, centrifuge at 300 g for 5 minutes to collect and count the cells; after counting, divide the cells into 5×10⁻⁶ cells per cell line. 6 Cells were evenly seeded into 6-well plates. When the cell confluence reached 80%, the cells were streaked vertically into the 6-well plates using a 200 μL pipette tip. The cells were washed 2-3 times with sterile 1×PBS to remove detached cells and cell debris. The cells were then cultured in serum-free medium containing afatinib (250 nM and 500 nM) and compound 13 (250 nM and 500 nM), respectively. The time was recorded as 0 hours. Subsequently, photos were taken at 12 hours and 13 hours, and the data were saved for statistical analysis.

[0093] The experimental results showed that, compared with the control drug (afatinib), compound 13 significantly inhibited cell migration in three types of human lung cancer cells with statistical significance (as shown in Figures 8-10).

[0094] Example 5: In vivo safety experiment Male C57 mice (Jiangsu Jicui Pharmaceutical Biotechnology Co., Ltd., strain name: C57BL / 6JGpt, strain number: N000013) aged 6-8 weeks and weighing approximately 22 grams were used. Five mice were divided into four groups: a normal control group (no treatment), a solvent control group (containing 3% DMSO, 2% propylene glycol, 2% ethanol, and 2.5% polyoxyethylene 40 hydrogenated castor oil), an afatinib group, and a compound 13 group. Each mouse was administered 100 μL of the corresponding drug intraperitoneally daily at a concentration of 10 mg / kg / day (based on mouse body weight) for 14 consecutive days, during which mouse body weight was monitored. On day 14 after drug administration, blood samples were collected from mice to detect liver function (ALT and AST) and kidney function (BUN and CRE) biochemical indicators (the kits were all purchased from Nanjing Jiancheng Bioengineering Institute Co., Ltd., with catalog numbers: ALT: C009-2-1, AST: C010-2-1, BUN: C013-2-1, CRE: C011-2-1). In addition, liver and kidney tissues were subjected to histopathological HE staining.

[0095] The results showed that, compared with the normal control group and the solvent control group, the experimental groups treated with compound 13 and afatinib had no significant effect on the body weight of the mice, and the mice's body weight showed an increasing trend, with the mice in good condition during the period. Biochemical tests showed no significant changes in any of the treated groups compared with the normal control and solvent control groups. Pathological HE staining showed no obvious abnormalities in the liver and kidney tissues. These results indicate that compound 13 has good in vivo safety (as shown in Figures 11-13).

[0096] Example 6: Evaluation of in vivo antitumor drug efficacy After BaF3 cells were cultured and expanded in vitro, they were resuspended in PBS buffer and counted. The cell concentration was adjusted to 8 × 10⁶ cells per 100 μL suspension. 5 Cells. The cell suspension was thoroughly mixed with an equal volume of Matrigel on ice to prepare a 200 μL cell-Matrigel (1:1) inoculum. Male BALB / c-Nude nude mice (Jiangsu Jicui Pharmaceutical Biotechnology Co., Ltd., strain number: D000521), aged 6-8 weeks and weighing 22 g, were slowly injected subcutaneously into the right scapular region of the mice. Each mouse was inoculated with 100 μL (containing 8 × 10⁶ cells). 5 (BaF3 cells). The mice's condition and tumor formation were observed regularly.

[0097] Once the transplanted tumor has grown to a palpable size and reached a stable volume of approximately 100 mm3 All tumor-forming mice were randomly divided into two groups of five each: one group served as the afatinib control group, and the other group served as the compound 13 experimental group. Both groups were administered the drug at a dose of 25 mg / kg / day (based on mouse body weight). The drug was dissolved in a prepared sterile carrier solution and injected intratumorally, with each mouse receiving 50 μL of the drug once daily for 12 consecutive days. During treatment, the major axis (a) and minor axis (b) of the tumor were measured using electronic calipers, and the tumor volume was calculated using the formula V = 0.5 × a × b. 2 The tumor volume was calculated and recorded at each time point, while the changes, activity status and drug administration site reactions of the mice were closely observed and recorded.

[0098] In vivo results showed that, throughout the treatment period, the growth of subcutaneous xenograft volume in mice treated with compound 13 was significantly inhibited compared to the afatinib control group, and no significant treatment-related abnormalities were observed in the general condition of the mice. This indicates that compound 13 exhibits superior antitumor activity compared to afatinib in the BALB / C-Nude nude mouse BaF3 cell xenograft model (as shown in Figure 14).

[0099] The above results indicate that: (1) Breakthrough anti-drug resistance effect: It effectively overcomes the EGFR C797S triple mutation in both in vivo and in vitro models. Specifically, in vitro, compound 13 exhibits extremely strong inhibitory activity and pro-apoptotic ability against BaF3 cells carrying the EGFR Del19 / T790M / C797S triple mutation. In CCK-8 and apoptosis experiments, it showed significantly better effects than afatinib at very low concentrations (20-60 nM). Existing TKIs such as afatinib and osimertinib are completely ineffective against this type of mutation. More importantly, in the BaF3 EGFR-Del19-T790M-C797S cell xenograft tumor model in nude mice, compound 13 (25 mg / kg) showed significantly stronger tumor growth inhibition ability than afatinib (25 mg / kg) without causing significant treatment-related toxicity. This directly demonstrates that compound 13 can effectively overcome C797S-mediated ultimate drug resistance even under complex physiological conditions, extending its effects from in vitro to in vivo, and possessing clear translational medical value. The technical reason for this effect is that this breakthrough directly stems from the precise structural reconstruction of the 6-position substituent in the quinazoline core, specifically replacing the traditional acrylamide group with a chloroacetyl group. The C797S mutation causes the kinase to lose the cysteine ​​residue that undergoes Michael addition with the acrylamide warhead, but it does not affect its ability to undergo nucleophilic substitution with the chloroacetyl group as a nucleophile. Therefore, this invention, through a fundamental shift in the mechanism of action (from Michael addition to nucleophilic substitution), successfully bypasses the C797S drug resistance mechanism, providing a novel chemical solution to address the most challenging multidrug resistance problem in clinical practice.

[0100] (2) Significantly broadened antitumor spectrum: Effective against EGFR wild-type and non-EGFR-dependent tumors. Specifically, in EGFR wild-type A549 non-small cell lung cancer cells, compound 13 showed significantly superior activity to afatinib in proliferation inhibition, colony formation inhibition, and cell migration inhibition experiments. This demonstrates that its antitumor effect is independent of EGFR-driven mutations. The technical reason for this effect is that the chlorine atom, as a superior leaving group compared to the acrylamide double bond system, allows the chloroacetyl warhead to undergo efficient and controllable nucleophilic substitution reactions with a wider range of conserved cysteine ​​residues within the ATP binding cavity. This covalent inhibition mechanism based on nucleophilic substitution is less dependent on the target microenvironment, thus inhibiting a wider variety of kinase targets, thereby producing a strong inhibitory effect on tumor cells that do not solely depend on the classical EGFR signaling pathway (such as A549), overcoming the narrow inhibitory spectrum of existing drugs.

[0101] (3) Stronger cell proliferation inhibition and apoptosis-inducing ability. Specifically, in the CCK-8 assay, plate colony formation assay, and flow cytometry apoptosis assay, compound 13, at the same concentration, showed stronger cell proliferation inhibition and apoptosis-inducing ability than afatinib in all tested tumor cell lines. The technical reason for this effect is that the chloroacetyl warhead has higher reactivity and better pharmacokinetic properties, and its covalent bond formation with target kinase is more efficient and faster. This more efficient and stable covalent binding leads to more thorough and longer-lasting target inhibition, thereby exhibiting a stronger anti-proliferation and apoptosis-inducing effect at the cellular functional level.

[0102] (4) Excellent ability to inhibit cell migration. Specifically, scratch assay results show that compound 13 can more effectively inhibit the migration of tumor cells, which is superior to afatinib. Inhibition of migration is of great significance in preventing tumor invasion and metastasis. The technical reason for this effect is that it is a direct manifestation of the above-mentioned broad-spectrum kinase inhibitory ability. The migration process of tumor cells is synergistically regulated by multiple signaling pathways. By inhibiting a wider range of kinase targets, compound 13 may simultaneously block multiple signaling pathways driving cell migration (such as c-Met, Axl, etc.), thereby producing a stronger anti-migration phenotype.

[0103] (5) Good in vivo safety. Specifically, in a 14-day repeated-dose safety evaluation in healthy C57 mice, compound 13 at a dose of 10 mg / kg showed no significant adverse effects on mouse body weight, biochemical indicators of major organs (ALT, AST, BUN, and CRE), or histopathological morphology compared to the solvent control group and the afatinib group, demonstrating good tolerability. This safety data, combined with the fact that no significant toxic abnormalities were observed at the effective dose (25 mg / kg) in the xenograft model, together indicate that compound 13 has a wide therapeutic window, providing important safety evidence for its clinical application. The technical reason for this effect is that the shorter alkyl chain length and higher reactivity of the chloroacetyl warhead may reduce the risk of nonspecific covalent binding to irrelevant proteins in the in vivo circulation. Its highly effective nature allows it to take effect at lower therapeutic doses, contributing to a wider safety window.

[0104] This invention achieves a breakthrough in both mechanism of action and technical efficacy through a key structural innovation: introducing a chloroacetylamino warhead at the 6-position of the quinazoline core to obtain a chloroacetylamino warhead-based quinazoline kinase inhibitor. This inhibitor is creatively used in the preparation of drugs for the prevention and / or treatment of tumors. Its core value lies not only in validating its ability to overcome ultimate drug resistance and expand therapeutic applications at the cellular level, but also in confirming its significant inhibitory effect on the most challenging triple-mutant drug-resistant tumors in animal models through rigorous in vivo pharmacodynamic experiments.

[0105] Finally, it should be noted that the above content is only used to illustrate the technical solution of the present invention, and is not intended to limit the scope of protection of the present invention. Simple modifications or equivalent substitutions made by those skilled in the art to the technical solution of the present invention do not depart from the essence and scope of the technical solution of the present invention.

Claims

1. The use of a chloroacetyl warhead-based quinazoline kinase inhibitor in the preparation of medicaments for the prevention and / or treatment of tumors, characterized in that, The chloroacetyl warhead-based quinazoline kinase inhibitor is selected from at least one of compound 13, a stereoisomer of compound 13, a solvate of compound 13, a pharmaceutically acceptable salt of compound 13, or a prodrug of compound 13; Compound 13 is 4-(4-fluoro-3-chlorophenylamino)-7-(S)tetrahydrofuran-3-yloxy)-6-(N-chloroacetyl)aminoquinazoline.

2. Use according to claim 1, characterized in that, The tumor is selected from at least one of breast cancer, lung cancer, kidney cancer, prostate cancer, colorectal cancer, and pancreatic cancer.

3. Use according to claim 2, characterized in that, The lung cancers mentioned include non-small cell lung cancer.

4. Use according to claim 3, characterized in that, The non-small cell lung cancer is mediated by EGFR.

5. The use according to claim 4, characterized in that, The non-small cell lung cancer includes tumors resistant to at least one of the first-generation, second-generation, and third-generation EGFR tyrosine kinase inhibitors.

6. The use according to claim 5, characterized in that, The first-generation EGFR tyrosine kinase inhibitor is selected from at least one of gefitinib, erlotinib, and icotinib; and / or The second-generation EGFR tyrosine kinase inhibitor is selected from at least one of afatinib and dacomitinib; and / or The third-generation EGFR tyrosine kinase inhibitor is selected from at least one of osimertinib, ametinib, and vormetinib.

7. Use according to claim 5, characterized in that, characterized in that, The non-small cell lung cancer is selected from those carrying at least one of the following: EGFR T790M, EGFR C797S, EGFR L858R, and EGFR 19del drug resistance mutations.

8. Use according to claim 4, characterized in that, The non-small cell lung cancer mentioned is EGFR wild-type non-small cell lung cancer.

9. Use according to claim 1, characterized in that, The drug contains a therapeutically effective amount of the chloroacetyl warhead-based quinazoline kinase inhibitor.

10. Use according to claim 9, characterized in that, Based on the patient's weight, the effective therapeutic dose is 0.1 mg / kg / day - 25 mg / kg / day.

11. Use according to claim 1, characterized in that, The drug also includes pharmaceutically acceptable carriers and / or excipients.

12. Use according to claim 1, characterized in that, The dosage form of the drug is tablets, capsules, injections, powder for injection, or inhalation.

13. A method for preparing a quinazoline kinase inhibitor based on a chloroacetyl warhead, characterized in that, The chloroacetyl warhead-based quinazoline kinase inhibitor is selected from at least one of compound 13, a stereoisomer of compound 13, a solvate of compound 13, a pharmaceutically acceptable salt of compound 13, or a prodrug of compound 13, wherein the preparation method of compound 13 includes the following steps: S1. 4-Chloro-6-nitro-7-fluoroquinazoline and 4-fluoro-3-chloroaniline are reacted in isopropanol with triethylamine and reacted at 50℃~70℃ for 5~7 hours to obtain 4-(4-fluoro-3-chlorophenylamino)-7-fluoro-6-nitroquinazoline; S2. The 4-(4-fluoro-3-chlorophenylamino)-7-fluoro-6-nitroquinazoline is reacted with S-(+)-3-hydroxytetrahydrofuran in a sodium hydride / tetrahydrofuran system at 90℃~110℃ for 7~9 hours to obtain 4-(4-fluoro-3-chlorophenylamino)-7-((S)tetrahydrofuran-3-yloxy)-6-nitroquinazoline; S3. The 4-(4-fluoro-3-chlorophenylamino)-7-((S)tetrahydrofuran-3-yloxy)-6-nitroquinazoline, reduced iron powder, and ammonium chloride are reacted in an ethanol-water mixed solvent at 80℃~100℃ for 5~7 hours to carry out a nitro reduction reaction, yielding 4-(4-fluoro-3-chlorophenylamino)-7-((S)tetrahydrofuran-3-yloxy)-6-aminoquinazoline; S4. Under ice bath conditions, a tetrahydrofuran solution of chloroacetyl chloride was added dropwise to the 4-(4-fluoro-3-chlorophenylamino)-7-((S)tetrahydrofuran-3-yloxy)-6-aminoquinazoline. The reaction was carried out at room temperature for 3 to 5 hours, and the mixture was purified by chromatography to obtain compound 13, which is 4-(4-fluoro-3-chlorophenylamino)-7-(S)tetrahydrofuran-3-yloxy)-6-(N-chloroacetyl)aminoquinazoline.

14. The method of claim 13, wherein, In step S1, the molar ratio of 4-chloro-6-nitro-7-fluoroquinazoline to 4-fluoro-3-chloroaniline is 1:1.0~1.2; In step S2, the molar ratio of 4-(4-fluoro-3-chlorophenylamino)-7-fluoro-6-nitroquinazoline to S-(+)-3-hydroxytetrahydrofuran is 1:1.1~1.2; In step S4, the molar ratio of 4-(4-fluoro-3-chlorophenylamino)-7-((S)tetrahydrofuran-3-yloxy)-6-aminoquinazoline to chloroacetyl chloride is 1:1.0~1.5.

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