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Significance Epstein–Barr virus (EBV)-associated diseases represent a major cause of morbidity worldwide, accounting for at least 1.5% of the global cancer burden, and contributing to autoimmunity. Viral reactivation from latency is associated with an increased risk of EBV-driven cancers, and thus may be a target for disease prevention. Anti-EBV agents that are safe, effective, and suitable for continuous long-term treatment are needed to address questions of whether suppression of lytic reactivation can reduce the incidence of EBV-associated diseases. Here we report that tenofovir disoproxil fumarate and tenofovir alafenamide, drugs with excellent safety profiles used for HIV prophylaxis, are potent inhibitors of EBV. This study identifies compounds that enable clinical investigations of antiviral therapy for EBV in efforts toward disease prevention.

Tenofovir prodrugs potently inhibit Epstein–Barr virus lytic DNA replication by targeting the viral DNA polymerase