TY - JOUR
T1 - Competitive Binding Assay with an Umbelliferone-Based Fluorescent Rexinoid for Retinoid X Receptor Ligand Screening
AU - Yamada, Shoya
AU - Kawasaki, Mayu
AU - Fujihara, Michiko
AU - Watanabe, Masaki
AU - Takamura, Yuta
AU - Takioku, Maho
AU - Nishioka, Hiromi
AU - Takeuchi, Yasuo
AU - Makishima, Makoto
AU - Motoyama, Tomoharu
AU - Ito, Sohei
AU - Tokiwa, Hiroaki
AU - Nakano, Shogo
AU - Kakuta, Hiroki
N1 - Funding Information:
The authors are grateful to the Division of Instrumental Analysis, Okayama University, for the NMR and MS measurements. X-ray data were collected at the synchrotron facilities of the Photon Factory (PF) using beamlines NE3A and NW12A (proposal No. 2016G102 and 2018G006). We would like to thank Dr. Tomoya Hirano (Osaka University of Pharmaceutical Sciences, Takatsuki, Osaka, Japan) for his helpful advice on quantum yield measurement. The authors thank Dr. Hiroyuki Kagechika (Tokyo Medical and Dental University, Tokyo, Japan), for providing PA452 and HX531. This work was partially supported by Grant-in-Aid from the Japan Society for the Promotion of Science (JSPS) 12J06716 (to S.Y.), JSPS KAKENHI Grant Numbers 16K18688, 18K14391 (to S.N.), and 17K06931(to S.I.), Okayama Foundation for Science and Technology (to H.K.), and The Tokyo Biochemical Research Foundation (TBRF) (to H.K.).
Publisher Copyright:
© 2019 American Chemical Society.
PY - 2019/10/10
Y1 - 2019/10/10
N2 - 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.
AB - 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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U2 - 10.1021/acs.jmedchem.9b00995
DO - 10.1021/acs.jmedchem.9b00995
M3 - Article
C2 - 31483660
AN - SCOPUS:85072934666
SN - 0022-2623
VL - 62
SP - 8809
EP - 8818
JO - Journal of Medicinal Chemistry
JF - Journal of Medicinal Chemistry
IS - 19
ER -
