Development of time resolved fluorescence resonance energy transfer-based assay for FXR antagonist discovery.

Abstract

FXR (farnesoid X receptor, NRIH4), a nuclear receptor, plays a major role in the control of cholesterol metabolism. FXR ligands have been investigated in preclinical studies for targeted therapy against metabolic diseases, but have shown limitations. Therefore, there is a need for new agonist or antagonist ligands of FXR, both for potential clinical applications, as well as to further elucidate its biological functions. Here we describe the use of the X-ray crystal structure of FXR complexed with the potent small molecule agonist GW4064 to design and synthesize a novel fluorescent, high-affinity probe (DY246) for time resolved fluorescence resonance energy transfer (TR-FRET) assays. We then used the TR-FRET assay for high throughput screening of a library of over 5000 bioactive compounds. From this library, we identified 13 compounds that act as putative FXR transcriptional antagonists.

• Authors: Donna D Yu, Wenwei Lin, Taosheng Chen, Barry M Forman
• Journal: Bioorganic & medicinal chemistry (Bioorg Med Chem) Vol. 21 Issue 14 Pg. 4266-78 (Jul 15 2013) ISSN: 1464-3391 [Electronic] England
• PMID: 23688559 (Publication Type: Journal Article, Research Support, N.I.H., Extramural)
• Copyright: Copyright © 2013 Elsevier Ltd. All rights reserved.

Chemical References

• DY246 compound
• Fluoresceins
• Fluorescent Dyes
• Isoxazoles
• Receptors, Cytoplasmic and Nuclear
• farnesoid X-activated receptor
• GW 4064

Topics

• Biological Assay (methods)
• Drug Discovery
• Fluoresceins (chemical synthesis, chemistry, pharmacology)
• Fluorescence Resonance Energy Transfer
• Fluorescent Dyes (chemical synthesis, chemistry, pharmacology)
• Gene Expression Regulation (drug effects)
• Isoxazoles (chemical synthesis, chemistry, pharmacology)
• Molecular Structure
• Protein Binding (drug effects)
• Receptors, Cytoplasmic and Nuclear (agonists, antagonists & inhibitors)