Abstract：
OBJECTIVE: The objective of this study is to identify dual-target inhibitors against EGFR/c-Met through virtual screening, dynamic simulation, and biological activity evaluation. This endeavor is aimed at overcoming the challenge of drug resistance induced by L858R/T790M mutants.
METHODS: Active structures were gathered to construct sets of drug molecules. Next, property filtering was applied to the drug structures within the compound library. Active compounds were then identified through virtual screening and cluster analysis. Subsequently, we conducted MTT antitumor activity evaluation and kinase inhibition assays for the active compounds to identify the most promising candidates. Furthermore, AO staining and JC-1 assays were performed on the selected compounds. Ultimately, the preferred compounds underwent molecular docking and molecular dynamics simulation with the EGFR and c-Met proteins, respectively.
RESULTS: The IC50 of T13074 was determined as 2.446 μM for EGFRL858R/T790M kinase and 7.401 nM for c-Met kinase, underscoring its potential in overcoming EGFRL858R/T790M resistance. Additionally, T13074 exhibited an IC50 of 1.93 μM on the H1975 cell. Results from AO staining and JC-1 assays indicated that T13074 induced tumor cell apoptosis in a concentration-dependent manner. Notably, the binding energy between T13074 and EGFR protein was found to be -90.329 ± 16.680 kJ/mol, while the binding energy with c-Met protein was -139.935 ± 17.414 kJ/mol.
CONCLUSIONS: T13074 exhibited outstanding antitumor activity both in vivo and in vitro, indicating its potential utility as a dual-target EGFR/c-Met inhibitor. This suggests its promising role in overcoming EGFR resistance induced by the L858R/T790M mutation.
METHODS: Active structures were gathered to construct sets of drug molecules. Next, property filtering was applied to the drug structures within the compound library. Active compounds were then identified through virtual screening and cluster analysis. Subsequently, we conducted MTT antitumor activity evaluation and kinase inhibition assays for the active compounds to identify the most promising candidates. Furthermore, AO staining and JC-1 assays were performed on the selected compounds. Ultimately, the preferred compounds underwent molecular docking and molecular dynamics simulation with the EGFR and c-Met proteins, respectively.
RESULTS: The IC50 of T13074 was determined as 2.446 μM for EGFRL858R/T790M kinase and 7.401 nM for c-Met kinase, underscoring its potential in overcoming EGFRL858R/T790M resistance. Additionally, T13074 exhibited an IC50 of 1.93 μM on the H1975 cell. Results from AO staining and JC-1 assays indicated that T13074 induced tumor cell apoptosis in a concentration-dependent manner. Notably, the binding energy between T13074 and EGFR protein was found to be -90.329 ± 16.680 kJ/mol, while the binding energy with c-Met protein was -139.935 ± 17.414 kJ/mol.
CONCLUSIONS: T13074 exhibited outstanding antitumor activity both in vivo and in vitro, indicating its potential utility as a dual-target EGFR/c-Met inhibitor. This suggests its promising role in overcoming EGFR resistance induced by the L858R/T790M mutation.
摘要：
目的：本研究的目的是通过虚拟筛选鉴定抗EGFR/c-Met的双靶点抑制剂，动态仿真,和生物活性评价。这一努力旨在克服由L858R/T790M突变体诱导的药物抗性的挑战。
方法：收集活性结构以构建药物分子组。接下来,将性质过滤应用于化合物库中的药物结构。然后通过虚拟筛选和聚类分析鉴定活性化合物。随后，我们对活性化合物进行了MTT抗肿瘤活性评估和激酶抑制试验,以确定最有希望的候选化合物.此外，对所选化合物进行AO染色和JC-1测定。最终,优选化合物与EGFR和c-Met蛋白进行分子对接和分子动力学模拟，分别。
结果：T13074的IC50对于EGFRL858R/T790M激酶为2.446μM，对于c-Met激酶为7.401nM，强调其在克服EGFRL858R/T790M抗性方面的潜力。此外,T13074对H1975细胞的IC50为1.93μM。AO染色和JC-1测定的结果表明T13074以浓度依赖性方式诱导肿瘤细胞凋亡。值得注意的是,发现T13074与EGFR蛋白之间的结合能为-90.329±16.680kJ/mol，与c-Met蛋白的结合能为-139.935±17.414kJ/mol。
结论：T13074在体内和体外均表现出优异的抗肿瘤活性，表明其作为双靶点EGFR/c-Met抑制剂的潜在效用。这表明其在克服由L858R/T790M突变诱导的EGFR抗性中的有希望的作用。
方法：收集活性结构以构建药物分子组。接下来,将性质过滤应用于化合物库中的药物结构。然后通过虚拟筛选和聚类分析鉴定活性化合物。随后，我们对活性化合物进行了MTT抗肿瘤活性评估和激酶抑制试验,以确定最有希望的候选化合物.此外，对所选化合物进行AO染色和JC-1测定。最终,优选化合物与EGFR和c-Met蛋白进行分子对接和分子动力学模拟，分别。
结果：T13074的IC50对于EGFRL858R/T790M激酶为2.446μM，对于c-Met激酶为7.401nM，强调其在克服EGFRL858R/T790M抗性方面的潜力。此外,T13074对H1975细胞的IC50为1.93μM。AO染色和JC-1测定的结果表明T13074以浓度依赖性方式诱导肿瘤细胞凋亡。值得注意的是,发现T13074与EGFR蛋白之间的结合能为-90.329±16.680kJ/mol，与c-Met蛋白的结合能为-139.935±17.414kJ/mol。
结论：T13074在体内和体外均表现出优异的抗肿瘤活性，表明其作为双靶点EGFR/c-Met抑制剂的潜在效用。这表明其在克服由L858R/T790M突变诱导的EGFR抗性中的有希望的作用。
