Berthier Alexandre, Staels Bart, Lefebvre Philippe
Univ. Lille, Inserm, CHU Lille, Institut Pasteur de Lille, U1011-EGID, 59000 Lille, France.
STAR Protoc. 2021 Jul 7;2(3):100658. doi: 10.1016/j.xpro.2021.100658. eCollection 2021 Sep 17.
Here, we describe an optimized protocol to identify specific nuclear receptor ligands. First, to rule out any compound interference with luciferase activity per se, we describe an assay assessing potential inhibition or activation of luciferase enzymatic activity. Second, to comply with EMA and FDA guidelines to mitigate drug-drug interactions, we detail assays assessing constitutive androstane receptor (CAR) and pregnane X receptor (PXR) activation ability. Finally, to minimize off-target detection effects, we describe the use of mammalian one- (or two-) hybrid systems. For complete details on the use and execution of this protocol, please refer to Hering et al. (2018).
在此，我们描述了一种用于鉴定特定核受体配体的优化方案。首先，为排除任何化合物对荧光素酶活性本身的干扰，我们描述了一种评估荧光素酶酶活性潜在抑制或激活的测定方法。其次，为符合欧洲药品管理局（EMA）和美国食品药品监督管理局（FDA）关于减轻药物相互作用的指南，我们详细介绍了评估组成型雄烷受体（CAR）和孕烷X受体（PXR）激活能力的测定方法。最后，为尽量减少脱靶检测效应，我们描述了哺乳动物单（或双）杂交系统的使用。有关该方案使用和执行的完整详细信息，请参考赫林等人（2018年）的文献。
