Potent, selective and cell penetrant inhibitors of SF-1 by functional ultra-high-throughput screening
Potent, selective and cell penetrant inhibitors of SF-1 by functional ultra-high-throughput screening
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DOI：
10.1124/mol.108.045963
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发表时间：
2008-06-01
影响因子：
3.6
通讯作者：
Hodder, Peter
中科院分区：
文献类型：
--
作者：
Madoux, Franck;Li, Xiaolin;Hodder, Peter
类固醇生成因子1（SF-1，也称为NR 5A 1）是属于核受体超家族的转录因子。虽然该家族的大多数成员已被广泛表征，但SF-1的治疗潜力和药理学仍然难以捉摸。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。在使用嵌合SF-1构建体的基于反式激活细胞的测定中筛选来自国立卫生研究院分子库小分子储存库的一组64，908种化合物。两个类似的异喹啉酮，乙基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]氧基丙酸酯（SID 7969543）和2-[2-[2-（2-甲氧基苯基）氨基]乙基]-2-（三氟甲基）苯甲酰胺（1，3-苯并间二氧杂环戊烯-5-基甲基氨基）-2-氧代乙基]-1-氧代异喹啉-5-基]氧基丙酸酯和（SID 7970631）被鉴定为有效的亚微摩尔抑制剂，产生760和260 nM的IC 50值。这些化合物在采用全长SF-1蛋白及其天然反应元件的更具生理功能的测定中保留了其效力，分别产生30和16 nM的IC 50值。通过对RAR相关孤儿受体A（RORA）、单纯疱疹病毒转录激活蛋白Vmw 65（VP 16）和肝受体同源物1（LRH-1）进行基于反式激活的功能测定，证实了这些异喹啉酮的选择性。测定了它们的细胞毒性、溶解性、渗透性和代谢稳定性。这些异喹啉酮代表了研究SF-1治疗潜力的有价值的化学探针。
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 IC50 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 IC50 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.