Development and implementation of a cell-based assay to discover agonists of the nuclear receptor REV-ERBα
Development and implementation of a cell-based assay to discover agonists of the nuclear receptor REV-ERBα
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DOI：
10.14440/jbm.2018.244
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发表时间：
2018-01-01
期刊：
影响因子：
--
通讯作者：
Gul, Sheraz
中科院分区：
文献类型：
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作者：
Hering, Yuliya;Berthier, Alexandre;Gul, Sheraz
核受体是参与调节多种生理过程的转录因子，其活性可以通过与相关小分子配体结合来调节。它们的功能障碍已被证明在糖尿病，癌症，炎症性疾病和激素抵抗疾病等疾病状态中发挥作用，这使它们成为药物发现的有趣目标。核受体REV-ERB α参与调节昼夜节律和代谢。它的天然配体是血红素，并且在鉴定新型合成调节剂以用作表征其功能的工具和用作治疗代谢紊乱的药物方面存在显著的兴趣。为此，我们建立了一种基于哺乳动物细胞的双杂交测定系统，能够测量REV-ERB α与其辅阻遏物核辅阻遏物1之间的相互作用。根据工业标准验证了该测定，并用于筛选LOPAC（R）（1280）文库的子集和来自不同化合物文库的29568种化合物。在一组计数器和选择性测定中对主要命中物的分析证实，REV-ERB α活性可以被调节，并且已经鉴定了用于优化的化学支架。
The nuclear receptors are transcription factors involved in the regulation of a variety of physiological processes whose activity can be modulated by binding to relevant small molecule ligands. Their dysfunction has been shown to play a role in disease states such as diabetes, cancer, inflammatory diseases, and hormonal resistance ailments, which makes them interesting targets for drug discovery. The nuclear receptor REV-ERB alpha is involved in regulating the circadian rhythm and metabolism. Its natural ligand is heme and there is significant interest in identifying novel synthetic modulators to serve as tools to characterize its function and to serve as drugs in treating metabolic disorders. To do so, we established a mammalian cell-based two-hybrid assay system capable of measuring the interaction between REV-ERB alpha and its co-repressor, nuclear co-repressor 1. This assay was validated to industry standard criteria and was used to screen a subset of the LOPAC (R)(1280) library and 29568 compounds from a diverse compound library. Profiling of the primary hits in a panel of counter and selectivity assays confirmed that REV-ERB alpha activity can be modulated pharmacologically and chemical scaffolds have been identified for optimization.