title: High-throughput screening identifies a bisphenol inhibitor of SV40 large T antigen ATPase activity.
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.
authors.affiliation: Department of Biological Sciences, University of Pittsburgh, Pittsburgh, PA, USA.
authors.forename: Sandlin P
authors.initials: SP
authors.lastname: Seguin
authors.forename: Carrie W
authors.initials: CW
authors.lastname: Evans
authors.forename: Miranda
authors.initials: M
authors.lastname: Nebane-Akah
authors.forename: Sara
authors.initials: S
authors.lastname: McKellip
authors.forename: Subramaniam
authors.initials: S
authors.lastname: Ananthan
authors.forename: Nichole A
authors.initials: NA
authors.lastname: Tower
authors.forename: Melinda
authors.initials: M
authors.lastname: Sosa
authors.forename: Lynn
authors.initials: L
authors.lastname: Rasmussen
authors.forename: E Lucile
authors.initials: EL
authors.lastname: White
authors.forename: Brooks E
authors.initials: BE
authors.lastname: Maki
authors.forename: Daljit S
authors.initials: DS
authors.lastname: Matharu
authors.forename: Jennifer E
authors.initials: JE
authors.lastname: Golden
authors.forename: Jeffrey
authors.initials: J
authors.lastname: Aubé
authors.forename: Jeffrey L
authors.initials: JL
authors.lastname: Brodsky
authors.forename: James W
authors.initials: JW
authors.lastname: Noah
mesh_terms: D000251:Adenosine Triphosphatases / Q000037:antagonists & inhibitors*; D000818:Animals; D000952:Antigens, Polyomavirus Transforming / Q000378:metabolism*; D000998:Antiviral Agents / Q000494:pharmacology; D001559:Benzhydryl Compounds; D002460:Cell Line; D002522:Chlorocebus aethiops; D004285:Dogs; D004353:Drug Evaluation, Preclinical; D004791:Enzyme Inhibitors / Q000494:pharmacology; D057166:High-Throughput Screening Assays / Q000379:methods*; D010636:Phenols / Q000494:pharmacology*; D011120:Polyomavirus / Q000201:enzymology; D054852:Small Molecule Libraries / Q000032:analysis; D000098826:Bisphenol A Compounds
chemical_list: D000251:Adenosine Triphosphatases; D000952:Antigens, Polyomavirus Transforming; D000998:Antiviral Agents; D001559:Benzhydryl Compounds; D004791:Enzyme Inhibitors; D010636:Phenols; D054852:Small Molecule Libraries; C006780:bisphenol A; D000098826:Bisphenol A Compounds