Genistein inhibits the replication of enterovirus A71

Background Genistein, an isoflavone abundant in soybeans and other legumes, has shown antiviral activity against several viruses. Its effects on enteroviruses, particularly EV71—the causative agent of hand, foot, and mouth disease (HFMD)—remain incompletely understood. Methods In vitro assays were used to evaluate replication inhibition of EV71, assess mechanistic effects, and determine the compound’s broad-spectrum activity against other enteroviruses (EV68 and CA6). In vivo experiments in neonatal mice were conducted to further elucidate the role of genistein in EV71 replication. Results Genistein inhibited EV71 replication, as evidenced by reduced viral genome copies, decreased viral protein expression, and lower virion numbers. The compound also reduced replication of EV68 and CA6. Mechanistically, autophagy inhibition contributed to the suppression of EV71 replication, and restoring autophagy attenuated this effect. Additionally, genistein caused a G2/M cell cycle arrest, contributing to impaired EV71 replication. In neonatal mice, genistein conferred protection against EV71-associated disease. Conclusion The compound effectively suppresses EV71 replication and mitigates EV71-induced pathology, with concurrent activity against EV68 and CA6, and involves autophagy modulation and cell cycle disruption as part of its antiviral mechanism.