Madoux Franck, Li Xiaolin, Chase Peter, Zastrow Gina, Cameron Michael D, Conkright Juliana J, Griffin Patrick R, Thacher Scott, Hodder Peter
Scripps Research Molecular Screening Center, The Scripps Research Institute, Scripps Florida, Jupiter, Florida 33458, USA.
Mol Pharmacol. 2008 Jun;73(6):1776-84. doi: 10.1124/mol.108.045963. Epub 2008 Mar 11.
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.
类固醇生成因子1（SF-1，也称为NR5A1）是一种属于核受体超家族的转录因子。尽管该家族的大多数成员已得到广泛研究，但其治疗潜力和药理学特性仍不清楚。本文描述了通过合理的超高通量筛选（uHTS）策略鉴定和表征SF-1的选择性抑制化学探针的过程。在美国国立卫生研究院分子文库小分子储存库中的一组64908种化合物，在基于细胞的反式激活测定中使用嵌合SF-1构建体进行筛选。两种类似的异喹啉酮，2-[2-[2-(2,3-二氢-1,4-苯并二恶英-7-基氨基)-2-氧代乙基]-1-氧代异喹啉-5-基]氧基丙酸乙酯（SID7969543）和2-[2-[2-(1,3-苯并二恶唑-5-基甲基氨基)-2-氧代乙基]-1-氧代异喹啉-5-基]氧基丙酸乙酯（SID7970631），被鉴定为有效的亚微摩尔抑制剂，IC50值分别为760和260 nM。在使用全长SF-1蛋白及其天然反应元件的更生理功能测定中，这些化合物保持了其效力，IC50值分别为30和16 nM。通过基于反式激活的功能测定，对维甲酸相关孤儿受体A（RORA）、单纯疱疹病毒转录激活蛋白Vmw65（VP16）和肝脏受体同源物1（LRH-1），证实了这些异喹啉酮的选择性。还测量了它们的细胞毒性、溶解度、渗透性和代谢稳定性。这些异喹啉酮代表了用于研究SF-1治疗潜力的有价值的化学探针。
