Development of a complex scintillation proximity assay for high-throughput screening of PPARγ modulators

• 来源期刊: 中国药理学报（英文版）
• 关键词: peroxisome proliferator-activated receptor gamma retinoid X receptor alpha scintillation proximity assay high-throughput screening
• 年，卷（期）: 2005,（3）
• 页码范围: 339-344
• 页数: 6页
• 语种: 英文

摘要

Aim: To develop a complex high-throughput screening (HTS) assay based on scintillation proximity assay (SPA) technology for identification of novel peroxisome proliferator-activated receptor gamma (PPARγ) modulators. Methods: Fulllength PPARγ and retinoid X receptor alpha (RXRα), biotinylated PPAR response element (PPRE), [3H]BRL49653 and streptavidin-coated FlashPlate or microbead method was validated against conventional hydroxyapatite (HA) assay and applied to large-scale screening of 16 000 synthetic compounds and natural product extracts. Results: (1) IC50 values of positive control compounds (BRL49653 and rable and consistent with those reported elsewhere; (2) Approximately 178 compounds, showing more than 70% competitive inhibition on BRL49653 bindSecondary screening using FlashPlate and cross-reactivity studies with RARα, β,γ and RXRα, β, γ confirmed that 12 compounds possessed specific PPARγ binding properties including 2 with IC50 values less than 0.5 μmol/L and novel chemimicrobead, is a highly efficient, automatable, and robust tool to screen potential PPARγ modulators in HTS setting. Its application may be expanded to other nuclear receptors that form heterodimers upon activation.