Genetics-guided screening and biological validation identify zeylenol as a promising EGFR inhibitor in glioma.

Introduction: To overcome the high attrition rates in glioma drug discovery, this study established a systematic "Genetics-to-Drug" pipeline aimed at identifying potential therapeutic targets for glioma through causal inference and discovering potent natural inhibitors. Methods: We initiated the study with a drug-target Mendelian Randomization (MR) analysis, leveraging quantitative trait loci (QTL) and large-scale genome-wide association studies (GWAS) data for all glioma, glioblastoma (GBM), and non-GBM subtypes. Based on the identified targets, we performed a virtual screening of 17,931 ZINC15 compounds utilizing LibDock, absorption, distribution, metabolism, and excretion (ADME) profiling, toxicity prediction by komputer assisted technology (TOPKAT), CDOCKER, and molecular dynamics (MD) simulations. The binding affinity of selected candidates was confirmed via biolayer interferometry (BLI), and the therapeutic efficacy of the identified small molecules were biologically validated through Results: The druggable MR screening of 140 genes identified EGFR as a key causal target across all glioma subtypes (GBM, non-GBM, and all glioma). Through a series of computer-aided techniques, two promising lead compounds, tremulacin (ZINC000004098458) and zeylenol (ZINC000229763735), were prioritized. Subsequent BLI assays validated a direct physical interaction between zeylenol and EGFR, and biological validation demonstrated that zeylenol treatment significantly suppressed GBM cell proliferation and migration while promoting apoptosis, supporting its therapeutic potential as a novel EGFR inhibitor. Discussion: This study identifies EGFR as a causally relevant therapeutic target in glioma and highlights tremulacin and zeylenol as promising natural EGFR inhibitors, warranting further preclinical development of these compounds as novel glioma therapeutics.