High-throughput screening identifies a bisphenol inhibitor of SV40 large T antigen ATPase activity.

Department of Biological Sciences, University of Pittsburgh, Pittsburgh, PA, USA.

Journal of biomolecular screening 2012 Feb

filter terms:

The authors conducted a high-throughput screening campaign for inhibitors of SV40 large T antigen ATPase activity to identify candidate antivirals that target the replication of polyomaviruses. The primary assay was adapted to 1536-well microplates and used to screen the National Institutes of Health Molecular Libraries Probe Centers Network library of 306 015 compounds. The primary screen had an Z value of ~0.68, signal/background = 3, and a high (5%) DMSO tolerance. Two counterscreens and two secondary assays were used to prioritize hits by EC(50), cytotoxicity, target specificity, and off-target effects. Hits that inhibited ATPase activity by >44% in the primary screen were tested in dose-response efficacy and eukaryotic cytotoxicity assays. After evaluation of hit cytotoxicity, drug likeness, promiscuity, and target specificity, three compounds were chosen for chemical optimization. Chemical optimization identified a class of bisphenols as the most effective biochemical inhibitors. Bisphenol A inhibited SV40 large T antigen ATPase activity with an IC(50) of 41 µM in the primary assay and 6.2 µM in a cytoprotection assay. This compound class is suitable as probes for biochemical investigation of large T antigen ATPase activity, but because of their cytotoxicity, further optimization is necessary for their use in studying polyomavirus replication in vivo.

Citation

Sandlin P Seguin, Carrie W Evans, Miranda Nebane-Akah, Sara McKellip, Subramaniam Ananthan, Nichole A Tower, Melinda Sosa, Lynn Rasmussen, E Lucile White, Brooks E Maki, Daljit S Matharu, Jennifer E Golden, Jeffrey Aubé, Jeffrey L Brodsky, James W Noah. High-throughput screening identifies a bisphenol inhibitor of SV40 large T antigen ATPase activity. Journal of biomolecular screening. 2012 Feb;17(2):194-203