Probing the Stereochemical Requirements for Receptor Recognition of δ Opioid Agonists through Topographic Modifications in Position 1

A series of side-chain constrained tyrosine derivatives, 2‘,6‘-dimethyl-β-methyltyrosines (TMT), has been designed and incorporated into position 1 of the highly selective δ opioid agonists DPDPE (Tyr- d -Pen 2 -Gly-Phe- d -Pen 5 -OH) and deltorphin I (DELT I, Tyr- d -Ala-Phe-Asp-Val-Val-Gly-NH 2 ). Molecular mechanics calculations on isolated TMT residues and nuclear magnetic resonance (NMR) studies of the TMT 1 -containing peptides in DMSO showed that each of the four stereoisomers of TMT favors one particular rotamer of the side-chain χ 1 torsional angle. Therefore, substitution of four TMT isomers for Tyr 1 allows us to perform a systematic conformational scan through three staggered rotamers of the aromatic side chain, gauche (−), trans, and gauche (+), and to explore specific binding requirements of the receptor in relation to the side chain conformation. The potency and selectivity of four isomers of [TMT 1 ]DPDPE and four isomers of [TMT 1 ]DELT I were evaluated by radioreceptor binding assays in the rat brain using μ- and δ-selective radiolabeled ligands and by bioassays with guinea pig ileum (GPI, μ receptor) and mouse vas deferens (MVD, δ receptor). In the DPDPE series only one isomer, [(2 S,3 R )-TMT 1 ]DPDPE showed high potency and selectivity for the δ opioid receptors. The favorable side-chain rotamers found for this analogue, i.e., the trans rotamer of TMT 1 and the gauche (−) rotamer of Phe 4, were proposed as the most probable δ receptor-binding conformations of DPDPE analogues. Two [TMT 1 ]DELT I isomers possessed considerable δ receptor potencies. The (2 S,3 R )-TMT 1 isomer appeared to be a superpotent, but moderately δ-selective agonist, while the (2S,3S)-TMT 1 isomer showed the highest selectivity for the δ receptors in this series. Surprisingly, [(2 R,3 R )-TMT 1 ]DELT I also was moderately potent at the δ receptor. These results suggest that the δ receptor requirements for the linear DELT I analogues may be satisfied with two different modes of binding of the (2S,3S)- and (2 S,3 R )-TMT 1 isomers. This study provides important guidance for the design of peptide and non-peptide ligands selective for the δ opioid receptor.