High-Throughput Screening Identifies a Bisphenol Inhibitor of SV40 Large T Antigen ATPase Activity
| Autor: | Miranda Nebane-Akah, Sara McKellip, Melinda Sosa, Sandlin P. Seguin, James W. Noah, Jennifer E. Golden, Jeffrey Aubé, E. Lucile White, Nichole A. Tower, Daljit S. Matharu, Carrie W. Evans, Jeffrey L. Brodsky, Subramaniam Ananthan, Brooks E. Maki, Lynn Rasmussen |
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| Jazyk: | angličtina |
| Rok vydání: | 2011 |
| Předmět: |
SV40 large T antigen
Bisphenol viruses High-throughput screening Antigens Polyomavirus Transforming Drug Evaluation Preclinical Biology Biochemistry Antiviral Agents Article Analytical Chemistry Cell Line Small Molecule Libraries Dogs Phenols High-Throughput Screening Assays Chlorocebus aethiops Atpase activity Animals Large T-Antigen Benzhydryl Compounds Enzyme Inhibitors Adenosine Triphosphatases Virology Molecular biology Cell culture Molecular Medicine Polyomavirus Biotechnology |
| Popis: | 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 by44% 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. |
| Databáze: | OpenAIRE |
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