The devastating impact of porcine epidemic diarrhea virus (PEDV) on the global swine industry underscores an urgent need for effective antiviral therapies. Drug repurposing presents a promising strategy to accelerate the development of such treatments. In this study, we screened a custom‐designed library of 117 cholesterol‐lowering compounds for anti‐PEDV activity using a recombinant PEDV expressing enhanced green fluorescent protein (EGFP). Following two rounds of screening, four compounds exhibiting significant antiviral activity were identified. Among these, cetaben and digitonin displayed superior antiviral efficacy and higher selectivity indices. Subsequent dose–response analyses further confirmed that cetaben effectively suppresses viral replication in both classical and variant strains of PEDV. Time‐of‐addition assays revealed that cetaben exerts potent antiviral effects primarily at the preinfection stage by inhibiting viral internalization and syncytium formation. Notably, supplementation with exogenous cholesterol in infected cells completely abolished cetaben’s antiviral activity and restored viral infectivity, demonstrating that its anti‐PEDV mechanism is mediated through disruption of cellular cholesterol metabolism. Collectively, our findings identify cetaben, a cholesterol‐lowering agent, as a novel broad‐acting antiviral against PEDV with a host‐targeted mechanism of action. This study establishes a foundation for developing antiviral strategies that target cholesterol metabolism to combat PEDV and related coronaviruses.
Li et al. (Thu,) studied this question.