2. Academic Validation
  3. Development of an Affinity-Based Probe to Profile Endogenous Human Adenosine A3 Receptor Expression

Development of an Affinity-Based Probe to Profile Endogenous Human Adenosine A3 Receptor Expression

  • J Med Chem. 2023 Aug 24;66(16):11399-11413. doi: 10.1021/acs.jmedchem.3c00854.
Bert L H Beerkens 1,  Inge M Snijders 1,  Joep Snoeck 1,  Rongfang Liu 1,  Anton T J Tool 2,  Sylvia E Le Dévédec 1,  Willem Jespers 1,  Taco W Kuijpers 2 3,  Gerard J P van Westen 1,  Laura H Heitman 1 4,  Adriaan P IJzerman 1,  Daan van der Es 1
Affiliations

Affiliations

  • 1 Division of Drug Discovery and Safety, Leiden Academic Centre for Drug Research, Leiden University, Einsteinweg 55, 2333 CC Leiden, The Netherlands.
  • 2 Department of Molecular Hematology, Sanquin Research, Plesmalaan 125, 1066 CX Amsterdam, The Netherlands.
  • 3 Department of Pediatric Immunology, Rheumatology and Infectious Diseases, Emma Children's Hospital, Academic Medical Center, University of Amsterdam, Amsterdam, The Netherlands, Meibergdreef 9, 1105 AZ Amsterdam, The Netherlands.
  • 4 Oncode Institute, Einsteinweg 55, 2333 CC Leiden, The Netherlands.
PMID: 37531576 DOI: 10.1021/acs.jmedchem.3c00854
Abstract

The adenosine A3 receptor (A3AR) is a G protein-coupled receptor (GPCR) that exerts immunomodulatory effects in pathophysiological conditions such as inflammation and Cancer. Thus far, studies toward the downstream effects of A3AR activation have yielded contradictory results, thereby motivating the need for further investigations. Various chemical and biological tools have been developed for this purpose, ranging from fluorescent ligands to Antibodies. Nevertheless, these probes are limited by their reversible mode of binding, relatively large size, and often low specificity. Therefore, in this work, we have developed a clickable and covalent affinity-based probe (AfBP) to target the human A3AR. Herein, we show validation of the synthesized AfBP in radioligand displacement, SDS-PAGE, and confocal microscopy experiments as well as utilization of the AfBP for the detection of endogenous A3AR expression in flow cytometry experiments. Ultimately, this AfBP will aid future studies toward the expression and function of the A3AR in pathologies.

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