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.
Shoya Yamada  1  2 ,  Mayu Kawasaki  3 ,  Michiko Fujihara  1  4 ,  Masaki Watanabe  1 ,  Yuta Takamura  1 ,  Maho Takioku  1 ,  Hiromi Nishioka  1 ,  Yasuo Takeuchi  1 ,  Makoto Makishima  5 ,  Tomoharu Motoyama  3 ,  Sohei Ito  3 ,  Hiroaki Tokiwa ,  Shogo Nakano  3 ,  Hiroki Kakuta  1
Affiliations
  • 1. Division of Pharmaceutical Sciences , Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences , 1-1-1, Tsushima-naka , Kita-ku, Okayama 700-8530 , Japan.
  • 2. Research Fellowship Division , Japan Society for the Promotion of Science , Sumitomo-Ichibancho FS Bldg., 8 Ichibancho , Chiyoda-ku, Tokyo 102-8472 , Japan.
  • 3. Graduate School of Integrated Pharmaceutical and Nutritional Sciences , University of Shizuoka , 52-1 Yada , Suruga-ku, Shizuoka 422-8526 , Japan.
  • 4. AIBIOS Co. Ltd. , Tri-Seven Roppongi 8F 7-7-7 Roppongi , Minato-ku, Tokyo 106-0032 Japan.
  • 5. Division of Biochemistry, Department of Biomedical Sciences , Nihon University School of Medicine , 30-1 Oyaguchi-kamicho , Itabashi-ku, Tokyo 173-8610 , Japan.
PMID: 31483660 DOI: 10.1021/acs.jmedchem.9b00995
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.

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