Development of novel fluorescent histamine H1-receptor antagonists to study ligand-binding kinetics in living cells

Leigh A. Stoddart, Andrea J. Vernall, Monica Bouzo-Lorenzo, Reggie Bosma, Albert J. Kooistra, Chris De Graaf, Henry F. Vischer, Rob Leurs, Stephen J. Briddon, Barrie Kellam, Stephen J. Hill*

*Corresponding author for this work

Research output: Contribution to JournalArticleAcademicpeer-review

Abstract

The histamine H1-receptor (H1R) is an important mediator of allergy and inflammation. H1R antagonists have particular clinical utility in allergic rhinitis and urticaria. Here we have developed six novel fluorescent probes for this receptor that are very effective for high resolution confocal imaging, alongside bioluminescence resonance energy transfer approaches to monitor H1R ligand binding kinetics in living cells. The latter technology exploits the opportunities provided by the recently described bright bioluminescent protein NanoLuc when it is fused to the N-terminus of a receptor. Two different pharmacophores (mepyramine or the fragment VUF13816) were used to generate fluorescent H1R antagonists conjugated via peptide linkers to the fluorophore BODIPY630/650. Kinetic properties of the probes showed wide variation, with the VUF13816 analogues having much longer H1R residence times relative to their mepyramine-based counterparts. The kinetics of these fluorescent ligands could also be monitored in membrane preparations providing new opportunities for future drug discovery applications.

Original languageEnglish
Article number1572
JournalScientific Reports
Volume8
Issue number1
DOIs
Publication statusPublished - 1 Dec 2018

Funding

This work was supported by the Medical Research Council [grant number MR/N020081/1]; and the Innovative Medicines Initiative Joint Undertaking under grant agreement 115366, resources of which are composed of financial contribution from the European Union’s Seventh Framework Programme (FP7/2007-2013) and European Federation of Pharmaceutical Industries and Associations (EFPIA) companies’ in-kind contribution.

FundersFunder number
FP7/2007
European Federation of Pharmaceutical Industries and Associations
Medical Research CouncilMR/N020081/1
Seventh Framework Programme
Innovative Medicines Initiative115366

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