Eupalinolide B exerts cytotoxic effects against KRAS-mutant NSCLC through Nrf2/HO-1-regulated ferroptosis

Ferroptosis, an iron-dependent form of regulated cell death, is characterized by excessive reactive oxygen species (ROS) accumulation and lipid peroxidation of polyunsaturated fatty acids (PUFAs) in cellular membranes. Non-small cell lung cancer (NSCLC) harboring KRAS mutations often exhibits therapeutic resistance but may display high susceptibility to ferroptosis. Eupalinolide B (EB), a natural compound with documented anti-cancer activity, has not been thoroughly explored for its ferroptosis-inducing potential in KRAS-mutant NSCLC. In this study, we demonstrate that EB treatment significantly elevates ROS levels, intracellular iron accumulation, and lipid peroxidation in KRAS-mutant NSCLC cells, resulting in ferroptotic cell death. Molecular docking and cellular thermal shift assays reveal that EB directly binds to and activates heme oxygenase-1 (HO-1), a critical component of the Kelch-like ECH-associated protein 1 (Keap1)-Nrf2/HO-1 oxidative stress response pathway. Genetic or pharmacological inhibition of HO-1 attenuates EB-induced ferroptosis, underscoring the pivotal role of HO-1-mediated oxidative stress in this process. Furthermore, <i>in vivo</i> studies using KRAS-mutant H358 xenograft models confirm the potent anti-tumor effects of EB. Collectively, our findings establish that EB triggers ferroptosis in KRAS-mutant NSCLC by activating the Keap1-Nrf2/HO-1 pathway, suggesting a promising therapeutic strategy for this challenging malignancy.