Competitive Binding Assay with an Umbelliferone-Based Fluorescent Rexinoid for Retinoid X Receptor Ligand Screening

J Med Chem. 2019 Oct 10;62(19):8809-8818. doi: 10.1021/acs.jmedchem.9b00995. Epub 2019 Sep 18.

Abstract

Ligands for retinoid X receptors (RXRs), "rexinoids", are attracting interest as candidates for therapy of type 2 diabetes and Alzheimer's and Parkinson's diseases. However, current screening methods for rexinoids are slow and require special apparatus or facilities. Here, we created 7-hydroxy-2-oxo-6-(3,5,5,8,8-pentamethyl-5,6,7,8-tetrahydronaphthalen-2-yl)-2H-chromene-3-carboxylic acid (10, CU-6PMN) as a new fluorescent RXR agonist and developed a screening system of rexinoids using 10. Compound 10 was designed based on the fact that umbelliferone emits strong fluorescence in a hydrophilic environment, but the fluorescence intensity decreases in hydrophobic environments such as the interior of proteins. The developed assay using 10 enabled screening of rexinoids to be performed easily within a few hours by monitoring changes of fluorescence intensity with widely available fluorescence microplate readers, without the need for processes such as filtration.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Binding, Competitive
  • Fluorescent Dyes / chemistry*
  • Genes, Reporter
  • Humans
  • Hydrophobic and Hydrophilic Interactions
  • Inhibitory Concentration 50
  • Ligands*
  • Protein Binding
  • Retinoid X Receptors / agonists*
  • Retinoid X Receptors / genetics
  • Retinoid X Receptors / metabolism
  • Tetrahydronaphthalenes / chemistry
  • Umbelliferones / chemistry*
  • Umbelliferones / metabolism

Substances

  • Fluorescent Dyes
  • Ligands
  • Retinoid X Receptors
  • Tetrahydronaphthalenes
  • Umbelliferones