Hering Yuliya, Berthier Alexandre, Duez Helene, Lefebvre Philippe, Deprez Benoit, Gribbon Philip, Wolf Markus, Reinshagen Jeanette, Halley Francoise, Hannemann Juliane, Böger Rainer, Staels Bart, Gul Sheraz
Fraunhofer Institute for Molecular Biology and Applied Ecology, ScreeningPort, Schnackenburgallee 114, D-22525 Hamburg, Germany.
University of Lille-EGID, CHU, Institut Pasteur de Lille, INSERM UMR 1011, 1 rue du Professeur Calmette, BP245, 59019 Lille, France.
J Biol Methods. 2018 Jun 25;5(3):e94. doi: 10.14440/jbm.2018.244. eCollection 2018.
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-ERBα 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-ERBα 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 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-ERBα activity can be modulated pharmacologically and chemical scaffolds have been identified for optimization.
核受体是参与多种生理过程调节的转录因子，其活性可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性疾病等疾病状态中起作用，这使得它们成为药物研发的有趣靶点。核受体REV-ERBα参与调节昼夜节律和新陈代谢。其天然配体是血红素，人们对鉴定新型合成调节剂有着浓厚兴趣，这些调节剂可作为表征其功能的工具，并作为治疗代谢紊乱的药物。为此，我们建立了一种基于哺乳动物细胞的双杂交检测系统，能够测量REV-ERBα与其共抑制因子核共抑制因子1之间的相互作用。该检测方法已按照行业标准进行验证，并用于筛选LOPAC文库的一个子集以及来自不同化合物文库的29568种化合物。在一系列反筛选和选择性检测中对初步筛选出的活性化合物进行分析，证实REV-ERBα的活性可通过药理学方法进行调节，并且已确定了用于优化的化学骨架。
