Drug repurposing for pancreatic cancer: Lomitapide Mesylate as a potent HK2 inhibitor discovered via virtual screening.

Pancreatic cancer has an early diagnosis rate below 5% and is largely resistant to conventional chemotherapy. Aerobic glycolysis, a metabolic hallmark, drives Epithelial-Mesenchymal Transition (EMT) to promote tumor progression and drug resistance. Hexokinase 2 (HK2) is the rate-limiting enzyme that initiates glycolysis. It is specifically overexpressed in pancreatic cancer cells and is a potential therapeutic target. To identify HK2 inhibitors, we performed structure-based virtual screening of over 70,000 compounds. The top candidate, Lomitapide Mesylate (LM), was validated through molecular docking, molecular dynamics simulations, Drug Affinity Responsive Target Stability (DARTS), and Cellular Thermal Shift Assay (CETSA), confirming direct HK2 binding. In vitro, LM inhibited HK2 activity, exerting anti-proliferative and pro-apoptotic effects. In a mouse organoid-derived xenograft model, LM monotherapy suppressed tumor growth. Importantly, LM combined with gemcitabine significantly enhanced anti-tumor efficacy, indicating potential to reverse gemcitabine resistance. Mechanistically, this synergy was linked to LM-mediated reversal of HK2-driven EMT. Thus, LM inhibits HK2 to block EMT and synergize with gemcitabine, offering a novel therapeutic strategy.