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Background    Polygonum chinense  Linn. is a common medicinal plant in Southeast Asia and has been used in traditional medicine in Vietnam. The plant contains phytochemicals with various biological properties; however, its antiviral effect has not yet been demonstrated. This study was aimed to evaluate the anti-influenza virus activity of crude extracts of  P. chinense,  to characterize antiviral metabolites therefrom and to investigate their mechanisms of antiviral action.    Methods  The methanol (MeOH) extract and organic solvent layers of  P. chinense  were prepared by extraction and partition with relevant solvents. The ethyl acetate (EtOAc) layer showing antiviral activity was chromatographed repeatedly on SiO 2  and Sephadex LH-20 columns to give eight pure metabolites. Their chemical structures were determined by NMR and MS spectral data. Anti-influenza virus activity of the eight metabolites against virus strains A/Puerto Rico/8/34 (H1N1, PR8), A/Hong Kong/8/68 (H3N2, HK) and B/Lee/40 (Lee) was evaluated on the basis of cytopathic effect (CPE) and plaque inhibition assays. Time-of-addition, confocal microscopy and neuraminidase inhibition assay were performed for mode-of-action studies of active ingredients.    Results  The MeOH extract of  P. chinense  showed anti-influenza virus activity with EC 50  values ranging from 38.4 to 55.5 μg/mL in a CPE inhibition assay. Among the eight pure metabolites isolated from  P. chinense , ellagic acid ( PC5 ), methyl gallate ( PC7 ) and caffeic acid ( PC8 ) significantly inhibited viral replication in a dose-dependent manner in both plaque inhibition and CPE inhibition assays with EC 50  values ranging from 14.7 to 81.1 μg/mL and CC 50  values higher than 300 μg/mL. Mode-of-action studies suggested that  PC5  and  PC7  suppress virus entry into or replication in cells, while  PC8  targets influenza viral neuraminidase, even oseltamivir-resistant one.    Conclusion  These results demonstrated that  P. chinense  and its metabolites possess effective anti-influenza virus activities. The botanical materials of  P. chinense  could be a promising multitargeted inhibitor of influenza A and B viruses and applied to development of a novel herbal medicine.

Characterization and mechanisms of anti-influenza virus metabolites isolated from the Vietnamese medicinal plant Polygonum chinense