Ring size in cyclic endomorphin-2 analogs modulates receptor binding affinity and selectivityElectronic supplementary information (ESI) available. See DOI: 10.1039/c5ob00565e

The study reports the solid-phase synthesis and biological evaluation of a series of new side chain-to-side chain cyclized opioid peptide analogs of the general structure Tyr-[ d -Xaa-Phe-Phe-Asp]NH 2 , where Xaa = Lys ( 1 ), Orn ( 2 ), Dab ( 3 ), or Dap ( 4 ) (Dab = 2,4-diaminobutyric acid, Dap = 2,3-diaminopropionic acid), containing 17- to 14-membered rings. The influence of the ring size on binding to the MOP, DOP and KOP opioid receptors was studied. In general, the reduction of the size of the macrocyclic ring increased the selectivity for the MOP receptor. The cyclopeptide incorporating Xaa = Lys displayed subnanomolar MOP affinity but modest selectivity over the KOP receptor, while the analog with the Orn residue showed increased affinity and selectivity for MOP. The analog with Dab was a weak MOP agonist and did not bind to the other two opioid receptors. Finally, the peptide with Xaa = Dap was completely MOP receptor-selective with subnanomolar affinity. Interestingly, the deletion of one Phe residue from 1 led to the 14-membered Tyr-c[ d -Lys-Phe-Asp]NH 2 ( 5 ), a potent and selective MOP receptor ligand. The in vitro potencies of the new analogs were determined in a calcium mobilization assay performed in Chinese Hamster Ovary (CHO) cells expressing human recombinant opioid receptors and chimeric G proteins. A good correlation between binding and the functional test results was observed. The influence of the ring size, solid support and the N -terminal protecting group on the formation of cyclodimers was studied. A series of endomorphin-2 cyclic analogs with 17- to 14-membered rings was synthesized and evaluated in terms of opioid affinity and selectivity. Conformational studies enabled us to rationalize biological results.