¹⁹F-NMR quantification of drug efficacy exemplified with the human A_2A adenosine receptor

Nessa Pesaran Afsharian¹, Beining Jin¹, Kenneth Jacobson², Jens Carlsson³, Sunny Kim⁴, Zhan-Guo Gao², Matthew Eddy¹

Department of Chemistry, University of Florida, Gainesville, FL 32611, USA
2. Laboratory of Bioorganic Chemistry, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, MD 20892, USA
3. Department of Cell and Molecular Biology, Science for Life Laboratory, Uppsala University, Uppsala, Sweden
4. National Institute on Drug Abuse, National Institutes of Health, Bethesda, MD 20892-1013, USA

G protein-coupled receptors (GPCRs) are membrane sensory proteins that are targeted by more than 1/3 of FDA-approved drugs.  The magnitude of GPCR signaling spans a wide range and is determined by the efficacy of bound ligands.  Ligands known as partial agonists activate GPCRs to a sub-maximal level.  A potential benefit of partial agonists is the ability to fine-tune receptor signaling output.  However, partial agonist activity can be difficult to quantify in conventional cell-based signaling assays, especially for weak partial agonists. We used ¹⁹F-nuclear magnetic resonance (NMR) spectroscopy to quantify the efficacy of a small library of compounds that were predicted to elicit a range of signaling outputs for the human A_2Aadenosine receptor (A_2AAR), a representative class A GPCR.  Previous work from our lab established conditions for which ¹⁹F-NMR of A_2AAR quantitatively correlated with A_2AAR signaling outputs measured in cellular cyclic-AMP accumulation assays¹.  We apply these conditions to quantify the signaling output of a series of novel A_2AAR compounds and show that our earlier work can be extended to definitively determine drug efficacies.  Our results demonstrate the practical utility of ¹⁹F-NMR applied to GPCRs as an analytical tool that complements cell-based signaling assays.

References:

N. Thakur, A. P. Ray, B. Jin, N.P. Afsharian, E. Lyman, Z.-G. Gao, K. A. Jacobson, and M. T. Eddy, “Membrane Mimetic-Dependence of GPCR Energy Landscapes,” Structure (2024).

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