A high-throughput screen to identify potential inhibitors of HSV-2

Herpes simplex virus 2 (HSV-2) is a widespread sexually transmitted infection in the human population. Infection by HSV-2 is lifelong and causes periodic genital ulcers, which can lead to increased co-infection of human immunodeficiency virus (HIV). No clinical vaccine is available for HSV-2, and current antivirals used to suppress HSV-2 diseases have limited efficacy. Thus, there is a need to identify new antivirals against HSV-2. Given the lack of new treatment options, we established a cell-based high throughput screening (HTS) platform using an HSV-2 reporter virus and small molecule libraries to identify novel inhibitors of HSV-2. Our HTS achieved with a Z'- factor score of 0.86, which represents high robustness and reproducibility for this screen. We screened over 90,000 compounds, and ∼2500 small compounds were identified as potential inhibitors of HSV-2. From this set, 294 compounds were further analyzed in secondary assays, and we identified 2 potential inhibitors of HSV-2. These results indicate that we have developed a robust HTS to select for compounds that inhibit HSV gene expression in cell culture, which can lead to the discovery novel of inhibitors or host pathways involved in HSV-2 lytic infection.