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

• Published in: Journal of biomolecular screening Vol. 17; no. 2; pp. 194 - 203
• Main 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
• Format: Journal Article
• Language: English
• Published: United States 01.02.2012
• Subjects: Adenosine Triphosphatases - antagonists & inhibitors; Animals; Antigens, Polyomavirus Transforming - metabolism; Antiviral Agents - pharmacology; Benzhydryl Compounds; Cell Line; Cercopithecus aethiops; Dogs; Drug Evaluation, Preclinical; Enzyme Inhibitors - pharmacology; High-Throughput Screening Assays - methods; Phenols - pharmacology; Polyomavirus - enzymology; Small Molecule Libraries - analysis
• ISSN: 2472-5552; 1552-454X
• DOI: 10.1177/1087057111421630

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.