title: Potent, selective and cell penetrant inhibitors of SF-1 by functional ultra-high-throughput screening.
abstract: The steroidogenic factor 1 (SF-1, also known as NR5A1) is a transcription factor belonging to the nuclear receptor superfamily. Whereas most of the members of this family have been extensively characterized, the therapeutic potential and pharmacology of SF-1 still remains elusive. Described here is the identification and characterization of selective inhibitory chemical probes of SF-1 by a rational ultra-high-throughput screening (uHTS) strategy. A set of 64,908 compounds from the National Institute of Health's Molecular Libraries Small Molecule Repository was screened in a transactivation cell-based assay employing a chimeric SF-1 construct. Two analogous isoquinolinones, ethyl 2-[2-[2-(2,3-dihydro-1,4-benzodioxin-7-ylamino)-2-oxoethyl]-1-oxoisoquinolin-5-yl]oxypropanoate (SID7969543) and ethyl 2-[2-[2-(1,3-benzodioxol-5-ylmethylamino)-2-oxoethyl]-1-oxoisoquinolin-5-yl]oxypropanoate and (SID7970631), were identified as potent submicromolar inhibitors, yielding IC(50) values of 760 and 260 nM. The compounds retained their potency in a more physiologic functional assay employing the full-length SF-1 protein and its native response element, yielding IC(50) values of 30 and 16 nM, respectively. The selectivity of these isoquinolinones was confirmed via transactivation-based functional assays for RAR-related orphan receptor A (RORA), Herpes simplex virus transcriptional activator protein Vmw65 (VP16), and liver receptor homolog 1 (LRH-1). Their cytotoxicity, solubility, permeability and metabolic stability were also measured. These isoquinolinones represent valuable chemical probes to investigate the therapeutic potential of SF-1.
authors.affiliation: Scripps Research Molecular Screening Center, The Scripps Research Institute, Scripps Florida, Jupiter, Florida 33458, USA.
authors.forename: Franck
authors.initials: F
authors.lastname: Madoux
authors.forename: Xiaolin
authors.initials: X
authors.lastname: Li
authors.forename: Peter
authors.initials: P
authors.lastname: Chase
authors.forename: Gina
authors.initials: G
authors.lastname: Zastrow
authors.forename: Michael D
authors.initials: MD
authors.lastname: Cameron
authors.forename: Juliana J
authors.initials: JJ
authors.lastname: Conkright
authors.forename: Patrick R
authors.initials: PR
authors.lastname: Griffin
authors.forename: Scott
authors.initials: S
authors.lastname: Thacher
authors.forename: Peter
authors.initials: P
authors.lastname: Hodder
mesh_terms: D000818:Animals; D016466:CHO Cells; D006224:Cricetinae; D003412:Cricetulus; D004285:Dogs; D000882:Haplorhini; D006801:Humans; D007546:Isoquinolines / Q000737:chemistry / Q000494:pharmacology; D051379:Mice; D051381:Rats; D054852:Small Molecule Libraries / Q000737:chemistry / Q000494:pharmacology*; D054339:Steroidogenic Factor 1 / Q000037:antagonists & inhibitors* / Q000378:metabolism*
chemical_list: D007546:Isoquinolines; D054852:Small Molecule Libraries; D054339:Steroidogenic Factor 1