Abstract

Epidermal growth factor receptor (EGFR) is a key driver and therapeutic target in non-small cell lung cancer (NSCLC). However, the efficacy of EGFR tyrosine kinase inhibitors (EGFR-TKIs) is often limited by acquired resistance. Podophyllotoxin (PPT), a natural aryltetralin lignan, inhibits tubulin and moderately targets EGFR, but its clinical use is restricted by high toxicity and poor bioavailability. Given the roles of EGFR and tubulin in NSCLC progression and drug resistance, this study synthesized and evaluated PPT-derived triazole ester compounds for dual EGFR/tubulin targeting and anti-NSCLC activity. Two compound series were tested: PPT-2-propionyl triazole esters ( A1–1∼A1–10 ) and PPT-4-pentynoic acid triazole esters ( A2–1∼A2–10 ). The A1 series showed superior antitumor activity, with compound A1–3 exhibiting strong effects against A549, H460, and H1975 cells—most notably in A549 cells (IC₅₀ = 0.62 μM), representing a 5-fold improvement over PPT (IC₅₀ = 3.06 μM) and a 78-fold increase over gefitinib (IC₅₀ = 48.36 μM). A1–3 also showed about 20-fold lower cytotoxicity in normal BEAS-2B cells than PPT, indicating higher tumor selectivity. Mechanistic studies demonstrated that A1–3 inhibits colony formation, induces G₂/M arrest, suppresses migration, and promotes apoptosis in A549 cells. Molecular modeling confirmed stable binding to both EGFR and tubulin. Confocal microscopy revealed tubulin depolymerization, and Western blot analysis showed inhibition of EGFR activation and downstream AKT signaling, confirming dual-targeting action. In silico ADMET predictions suggest that A1–3 possesses favorable pharmacokinetic properties, supporting its potential as a lead compound. Collectively, A1–3 is a potent, low-toxicity dual EGFR/tubulin inhibitor in vitro , warranting further in vivo and preclinical investigation to fully assess its therapeutic potential for NSCLC.