In silico screening applied in drug discovery: as a novel fourth-generation EGFR inhibitor

Lung cancer is one of the most common malignancies and also the cancer with the highest mortality rate. Among them, non-small cell lung cancer (NSCLC) accounts for 86% of the total incidence of lung cancer. In recent years, small molecule inhibitors targeting the epidermal growth factor receptor (EGFR) have become a hot direction for treating NSCLC. Although EGFR inhibitors have developed to the fourth generation, resistance generated by third-generation EGFR inhibitors has not been solved effectively. Therefore, developing fourth-generation EGFR inhibitors with novel scaffolds to overcome resistance is urgently needed. This study used a drug discovery strategy with four parts: molecular filtering framework, ROC-guided virtual screening, clustering analysis and binding mode analysis. As a result, 13 compounds were obtained. What's more, cell and kinase tests were performed on the 13 hit compounds. Among them, the optimized compound T001-10026247 shows excellent inhibitory activity against H1975, A549, and H460 cells, with IC50 values of 0.26 +/- 0.01, 0.74 +/- 0.08 and 2.65 +/- 0.22 mu M, respectively. The IC50 values of T001-10026247 for EGFR(T790M/C797S/L858R) and EGFR(T790M/L858R) are 2.289 mu M and 1.614 mu M, respectively. In addition, in vitro and in vivo activity evaluation of the screened compound T001-10026247 was performed. The results suggested that compound T001-10026247 was a potential EGFR inhibitor after being validated by apoptosis, acridine orange (AO), JC-1, scratch and transwell experiments. Furthermore, the results of biological safety experiments showed that the toxicity of T001-10026247 was close to or even below the positive drug AZD9291. In summary, based on the drug discovery strategy, a novel fourth-generation EGFR inhibitor (T001-10026247) with a new scaffold was discovered.