Abstract Background Dengue fever continues to exert significant global impact, affecting populations worldwide with considerable public health and economic consequences. There is no antiviral drug for dengue. This study focuses on hypericin, a naturally occurring compound from Hypericum perforatum L. whose anti-dengue properties have been underexplored. We systematically examined its antiviral efficacy against dengue virus (DENV), revealing strong inhibitory effects and clarifying its precise antiviral mechanism. Methods The study assessed the efficacy of hypericin against DENV using various scientific methods like plaque assays and Western blotting. We looked into its antiviral mechanism. We used a time-of-addition approach during our research. Moreover, the basic mechanisms involved were studied through molecular docking, surface plasmon resonance (SPR), and co-immunoprecipitation (Co-IP). Results This study demonstrated that hypericin exhibits broad-spectrum antiviral activity against DENV-2 in cell lines derived from multiple species. In time-of-addition experiments, it showed inhibitory effects under co-treatment, direct virucidal, and post-treatment conditions. Crucially, hypericin primarily blocked viral attachment and entry stages, thereby effectively reducing intracellular viral load. Mechanistic investigations revealed a interaction between hypericin and the E protein, evidenced by a computational docking score of -7.0 kcal/mol and an experimental SPR-derived Kd of 7.18 µM. Furthermore, Co-IP assays demonstrated that hypericin competitively blocks the association between the E protein and its cellular receptor, HSP70. Conclusion As per these findings, the E protein was seen to be a target of hypericin with an antiviral activity against DENV-2 at multiple stages by limiting viral adsorption and viral entry projecting a molecular basis for the candidate molecule as a possible anti-dengue agent.
Qi et al. (2026) studied this question.