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Pharmacological characterization of DPTN and other selective A3 adenosine receptor antagonists

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Abstract

The A3 adenosine receptor (AR) is emerging as an attractive drug target. Antagonists are proposed for the potential treatment of glaucoma and asthma. However, currently available A3AR antagonists are potent in human and some large animals, but weak or inactive in mouse and rat. In this study, we re-synthesized a previously reported A3AR antagonist, DPTN, and evaluated its affinity and selectivity at human, mouse, and rat ARs. We showed that DPTN, indeed, is a potent A3AR antagonist for all three species tested, albeit a little less selective for mouse and rat A3AR in comparison to the human A3AR. DPTN’s Ki values at respective A1, A2A, A2B, and A3 receptors were (nM) 162, 121, 230, and 1.65 (human); 411, 830, 189, and 9.61 (mouse); and 333, 1147, 163, and 8.53 (rat). Its antagonist activity at both human and mouse A3ARs was confirmed in a cyclic AMP functional assay. Considering controversial use of currently commercially available A3AR antagonists in rats and mice, we also re-examined other commonly used and selective A3AR antagonists under the same experimental conditions. The Ki values of MRS1523 were shown to be 43.9, 349, and 216 nM at human, mouse, and rat A3ARs, respectively. MRS1191 and MRS1334 showed incomplete inhibition of [125I]I-AB-MECA binding to mouse and rat A3ARs, while potent human A3AR antagonists, MRS1220, MRE3008F20, PSB10, PSB-11, and VUF5574 were largely inactive. Thus, we demonstrated that DPTN and MRS1523 are among the only validated A3AR antagonists that can be possibly used (at an appropriate concentration) in mouse or rat to confirm an A3AR-related mechanism or function.

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The original data from this study will be made available upon reasonable request.

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Acknowledgements

We acknowledge support from Astrocyte Pharmaceuticals, Cambridge, MA. We thank Ryan Campbell and Harsha Rao (NIDDK, NIH) for technical assistance. We thank Dr. Bryan L. Roth (Univ. North Carolina at Chapel Hill) and National Institute of Mental Health’s Psychoactive Drug Screening Program (Contract # HHSN-271-2008-00025-C) for screening data.

Funding

National Institute of Diabetes and Digestive and Kidney Diseases, Intramural Research Program (grant no. ZIADK031117).

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  1. Molecular Recognition Section, Laboratory of Bioorganic Chemistry, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, MD, 20892-0810, USA

    Zhan-Guo Gao, R. Rama Suresh & Kenneth A. Jacobson

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  1. Zhan-Guo Gao
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Correspondence to Zhan-Guo Gao or Kenneth A. Jacobson.

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Gao, ZG., Suresh, R.R. & Jacobson, K.A. Pharmacological characterization of DPTN and other selective A3 adenosine receptor antagonists. Purinergic Signalling 17, 737–746 (2021). https://doi.org/10.1007/s11302-021-09823-5

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