title: A quantitative high-throughput screen identifies novel inhibitors of the interaction of thyroid receptor beta with a peptide of steroid receptor coactivator 2.
abstract: The thyroid hormone receptors (TR) are members of the nuclear hormone receptor (NHR) superfamily that regulate development, growth, and metabolism. Upon ligand binding, TR releases bound corepressors and recruits coactivators to modulate target gene expression. Steroid receptor coactivator 2 (SRC2) is an important coregulator that interacts with TRβ to activate gene transcription. To identify novel inhibitors of the TRβ and SRC2 interaction, the authors performed a quantitative high-throughput screen (qHTS) of a TRβ-SRC2 fluorescence polarization assay against more than 290 000 small molecules. The qHTS assayed compounds at 6 concentrations up to 92 µM to generate titration-response curves and determine the potency and efficacy of all compounds. The qHTS data set enabled the characterization of actives for structure-activity relationships as well as for potential artifacts such as fluorescence interference. Selected qHTS actives were tested in the screening assay using fluoroprobes labeled with Texas Red or fluorescein. The retest identified 19 series and 4 singletons as active in both assays with 40% or greater efficacy, free of compound interference, and not toxic to mammalian cells. Selected compounds were tested as independent samples, and a methylsulfonylnitrobenzoate series inhibited the TRβ-SRC2 interaction with 5 µM IC(50). This series represents a new class of thyroid hormone receptor-coactivator modulators.
authors.affiliation: NIH Chemical Genomics Center, National Human Genome Research Institute, National Institutes of Health, Bethesda, MD, USA. rjohnso2@mail.nih.gov
authors.forename: Ronald L
authors.initials: RL
authors.lastname: Johnson
authors.forename: Jong Yeon
authors.initials: JY
authors.lastname: Hwang
authors.forename: Leggy A
authors.initials: LA
authors.lastname: Arnold
authors.forename: Ruili
authors.initials: R
authors.lastname: Huang
authors.forename: Jennifer
authors.initials: J
authors.lastname: Wichterman
authors.forename: Indre
authors.initials: I
authors.lastname: Augustinaite
authors.forename: Christopher P
authors.initials: CP
authors.lastname: Austin
authors.forename: James
authors.initials: J
authors.lastname: Inglese
authors.forename: R Kiplin
authors.initials: RK
authors.lastname: Guy
authors.forename: Wenwei
authors.initials: W
authors.lastname: Huang
mesh_terms: D002478:Cells, Cultured; D004353:Drug Evaluation, Preclinical; D057166:High-Throughput Screening Assays; D006801:Humans; D051958:Nuclear Receptor Coactivator 2 / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000378:metabolism*; D010455:Peptides / Q000138:chemical synthesis / Q000378:metabolism*; D011485:Protein Binding / Q000187:drug effects; D054852:Small Molecule Libraries / Q000494:pharmacology; D037042:Thyroid Hormone Receptors beta / Q000037:antagonists & inhibitors / Q000378:metabolism*
chemical_list: D051958:Nuclear Receptor Coactivator 2; D010455:Peptides; D054852:Small Molecule Libraries; D037042:Thyroid Hormone Receptors beta