Evaluation of (S)-10-Hydroxycamptothecin Inhibitor of Herpes Simplex Type 1 Identified from Screening of a Library of Natural Products

Abstract

Background: Herpes simplex virus type 1 (HSV-1) causes serious illness in humans, especially in newborns and immunocompromised hosts. Public health requires the development of new, less toxic anti-HSV-1 drugs. Objectives: This study aimed to evaluate the potential anti-herpesvirus activity of natural products in an extensive library of 133 compounds by examining viral titers and the number of viral plaques. Methods: (S)-10-hydroxycamptothecin (10-HCPT) as an inhibitor against viral DNA replication in the lowest concentration ranges from a set of natural products consisting of screening 133 compounds. Each step of the viral replication cycle of HSV-1 on A549 cells was evaluated with different assays, including adsorption, penetration, time-of-addition assay, and quantitative polymerase chain reaction (PCR). The respective antiviral effects on HSV-1AN95 infection were assessed in vitro. Results: 10-HCPT was found to be a potent inhibitor of HSV-1 infection in the lowest concentration range from screening of a natural product library. The results showed that 10-HCPT significantly affects HSV-1 viral plaque formation inhibition, with a half maximal effective concentration (EC50) of 0.07 μM. The time of addition assay suggested that 10-HCPT had a viral inhibitory effect when added 8 hours after infection. It was further confirmed by reducing the expression of late viral genes including glycoprotein (g) and viral protein (VP) (gB, gD, gH, VP1/2, and VP16) 4 hours after infection in the 10-HCPT treatment group compared to positive controls by quantitative real-time PCR. The Western blotting results are inconsistent with other reported results. It showed that 10-HCPT did not affect gD and ICP4 during HSV-1 infection, and 10-HCPT appeared to affect other genes in the immediate-early (IE) and late (L) steps. Conclusions: 10-HCPT demonstrated anti-HSV activity on HSV-1. Their dose-dependent antiviral activity showed that specific cellular components might mediate their function rather than cytotoxicity. This survey suggests a new outlook in exploring effective treatment options for HSV-1 infections.