EV206 is a potent N14-allyl–substituted evodiamine derivative identified as a highly active heat shock protein 70 (Hsp70) inhibitor with significant anticancer efficacy in non-small cell lung cancer (NSCLC) models. In vitro, EV206 robustly suppresses proliferation across multiple NSCLC cell lines (H1299, A549, H460, H226B, PC9) with submicromolar IC₅₀ values (~0.20–0.46 µM), demonstrating ~10-fold greater potency than the parent compound evodiamine. It inhibits both anchorage-dependent and -independent colony formation and induces apoptosis via caspase-3 and PARP cleavage, while also reducing cancer stem cell phenotypes, including sphere formation and ALDH activity. Notably, EV206 retains equivalent activity in chemotherapy- and EGFR inhibitor–resistant cell lines. In vivo, intraperitoneal administration (10 mg/kg) significantly suppresses H460 xenograft tumor growth without measurable toxicity or body weight loss. Importantly, EV206 shows minimal cytotoxicity toward normal cells, highlighting a favorable therapeutic window and positioning it as a promising low-toxicity Hsp70-targeting anticancer lead compound.
