A versatile sub-nanomolar fluorescent ligand enables NanoBRET binding studies and single-molecule microscopy at the histamine H(3) receptor

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Item Type:	Article
Title:	A versatile sub-nanomolar fluorescent ligand enables NanoBRET binding studies and single-molecule microscopy at the histamine H(3) receptor
Creators Name:	Rosier, N., Grätz, L., Schihada, H., Möller, J., Işbilir, A., Humphrys, L.J., Nagl, M., Seibel, U., Lohse, M.J. and Pockes, S.
Abstract:	The histamine H(3) receptor (H(3)R) is considered an attractive drug target for various neurological diseases. We here report the synthesis of UR-NR266, a novel fluorescent H(3)R ligand. Broad pharmacological characterization revealed UR-NR266 as a sub-nanomolar compound at the H(3)R with an exceptional selectivity profile within the histamine receptor family. The presented neutral antagonist showed fast association to its target and complete dissociation in kinetic binding studies. Detailed characterization of standard H(3)R ligands in NanoBRET competition binding using UR-NR266 highlights its value as a versatile pharmacological tool to analyze future H(3)R ligands. The low nonspecific binding observed in all experiments could also be verified in TIRF and confocal microscopy. This fluorescent probe allows the highly specific analysis of native H(3)R in various assays ranging from optical high throughput technologies to biophysical analyses and single-molecule studies in its natural environment. An off-target screening at 14 receptors revealed UR-NR266 as a selective compound.
Keywords:	Binding Sites, Bioluminescence Resonance Energy Transfer Techniques, Drug Dose-Response Relationship, Fluorescent Dyes, HEK293 Cells, Histamine H3 Antagonists, Histamine H3 Receptors, Ligands, Molecular Structure, Single Molecule Imaging, Structure-Activity Relationship
Source:	Journal of Medicinal Chemistry
ISSN:	0022-2623
Publisher:	American Chemical Society
Volume:	64
Number:	15
Page Range:	11695-11708
Date:	12 August 2021
Official Publication:	https://doi.org/10.1021/acs.jmedchem.1c01089
PubMed:	View item in PubMed

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