Putranjivain A from Euphorbia jolkini inhibits both virus entry and late stage replication of herpes simplex virus type 2 in vitro

Aims: To investigate the in vitro antiviral properties of putranjivain A, isolated from the whole plant of Euphorbia jolkini Bioss (Euphorbiaceae). Methods and results: Herpes simplex virus (HSV)-2 strain 196-infected Vero cells were used. It was shown that putranjivain A exhibited antiviral activity with an IC50 of 7.9 +/- 1.2 muM using the XTT assay, with the IC50 value increasing with increasing multiplicity of infection. Using the plaque reduction assay, the IC50 and IC90 were 6.3 +/- 0.8 and 14.5 +/- 3.1 muM, respectively. Putranjivain A showed no cytotoxic effect on cell multiplication at concentrations that achieved antiviral activity. The 50% cell cytotoxic concentration (CC50) was 80.3 +/- 14.7 muM, and the selectivity index (SI) (ratio of CC50 to IC50) for the XTT and plaque reduction assays was 10.2 and 12.7, respectively. When tested for virucidal activity, putranjivain A significantly reduced viral infectivity at concentrations of 75 and 100 muM, but not at 50 muM or below. The results of the time-of-addition studies suggested that putranjivain A affected the late stage of HSV-2 replication at 25 muM. Interestingly, putranjivain A also showed inhibition of viral attachment and cell penetration. The combination of putranjivain A and aciclovir produced no interaction. Conclusions: Putranjivain A possesses antiviral activity, inhibiting viral attachment and penetration, and also interfering with the late stage of viral replication.