High-throughput screening identifies a bisphenol inhibitor of SV40 large T antigen ATPase activity |
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Authors: | Seguin Sandlin P Evans Carrie W Nebane-Akah Miranda McKellip Sara Ananthan Subramaniam Tower Nichole A Sosa Melinda Rasmussen Lynn White E Lucile Maki Brooks E Matharu Daljit S Golden Jennifer E Aubé Jeffrey Brodsky Jeffrey L Noah James W |
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Affiliation: | Department of Biological Sciences, University of Pittsburgh, Pittsburgh, PA, USA. |
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Abstract: | 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. |
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