Pharmacology of Kappa Opioid Receptors: Novel Assays and Ligands

The present study investigated the in vitro pharmacology of the human kappa Opioid Receptor using multiple assays, including calcium mobilization in cells expressing chimeric G proteins, the dynamic mass redistribution (DMR) label-free assay, and a bioluminescence resonance energy transfer (BRET) assay that allows measurement of receptor interaction with G protein and β-arrestin 2. In all assays, dynorphin A, U-69,593, and [D-Pro¹⁰]dyn(1-11)-NH₂ behaved as full agonists with the following rank order of potency [D-Pro¹⁰]dyn(1-11)-NH₂ > dynorphin A ≥ U-69,593. [Dmt¹,Tic²]dyn(1-11)-NH₂ behaved as a moderate potency pure antagonist in the kappa-β-arrestin 2 interaction assay and as low efficacy partial agonist in the Other assays. Norbinaltorphimine acted as a highly potent and pure antagonist in all assays except kappa-G protein interaction, where it displayed efficacy as an inverse agonist. The pharmacological actions of novel kappa ligands, namely the dynorphin A tetrameric derivative PWT2-Dyn A and the palmitoylated derivative Dyn A-palmitic, were also investigated. PWT2-Dyn A and Dyn A-palmitic mimicked dynorphin A effects in all assays showing similar maximal effects but 3-10 fold lower potency. In conclusion, in the present study, multiple in vitro assays for the kappa receptor have been set up and pharmacologically validated. In addition, PWT2-Dyn A and Dyn A-palmitic were characterized as potent full agonists; these compounds are worthy of further investigation in vivo for those conditions in which the activation of the kappa Opioid Receptor elicits beneficial effects e.g. pain and pruritus.

BRET; G protein-coupled receptor; PWT2-dyn A; biased agonism; calcium mobilization; dyn A-palmitic; kappa opioid receptor; label-free.