Novel Irreversible Agonists Acting at the A₁ Adenosine Receptor

The A₁ adenosine receptor (A₁AR) is an important G protein-coupled receptor that regulates a range of physiological functions. Herein we report the discovery of novel irreversible agonists acting at the A₁AR, which have the potential to serve as useful research tools for studying receptor structure and function. A series of novel adenosine derivatives bearing electrophilic substituents was synthesized, and four compounds, 8b, 15a, 15b, and 15d, were shown to possess similar potency and efficacy to the reference high efficacy agonist, NECA, in an assay of ERK1/2 phosphorylation assay. Insensitivity to antagonist addition in a real-time, label-free, xCELLigence assay was subsequently used to identify compounds that likely mediated their agonism through an irreversible interaction with the A₁AR. Of these compounds, 15b and 15d were more directly validated as irreversible agonists of the A₁AR using membrane-based [³H]­DPCPX and [³⁵S]­GTPγS binding experiments.