Abstract B136: In Silico Screening and Biological Evaluation of Traditional Chinese Medicines library against Switch-II Pocket of KRASG12D in pancreatic Cancer

Abstract

KRASG12D is a common cancer-driving mutation that fuels aggressive cancers, most notably in pancreatic, colorectal, and lung cancers. Although its role in disease progression is well established, treatment options that directly target KRASG12D are still limited. In this study, we employed a structure-based virtual screening approach to identify novel KRASG12D inhibitors. Molecular docking was implemented to screen a library of Traditional Chinese Medicine (TCM) compounds, followed by ADMET profiling. Top five drug-like hits were selected for 200 ns molecular dynamics (MD) simulations, and MM-GBSA binding free energy calculations. Furthermore, three compounds were selected for in vitro evaluation, among which Rubimaillin demonstrated potent anticancer activity with IC50 values of 1.10 ± 0.12 µM and 2.07 ± 0.02 µM against MIA PaCa-2 and PANC-1 cell lines, respectively, while exhibiting low toxicity in non-tumorous HEK-293 cells (CC50 = 38.92 ± 8.49 µM). Furthermore, Rubimaillin significantly inhibited cell migration and modulated key oncogenic biomarkers, with downregulation in KRAS protein levels, along with BCL2, SOX9, LC3 gene expression and upregulation in p53 gene expression. This study highlights Rubimaillin may serve as a potent KRASG12D inhibitor candidate for preclinical studies and demonstrates the effectiveness of integrating computational screening with natural product-based discovery.