title: Structure-activity studies on nociceptin analogues: ORL1 receptor binding and biological activity of cyclic disulfide-containing analogues of nociceptin peptides.
abstract: Nociceptin/Orphanin FQ is an endogenous peptide ligand for the opioid receptor-like 1 (ORL1) receptor. To investigate the structural and conformational requirements of the nociceptin (NC)-receptor interaction, six cyclic analogues containing Cys disulfide linkages were designed and synthesized. Analogues cyclized at the N-terminal part, cyclo[Cys(0), Cys(7)]NC(1-13)-NH(2) (2) and cyclo[Cys(0), Cys(11)]NC(1-13)-NH(2) (4), and their corresponding linear peptides had very low activities in both the receptor binding and the GTP gamma S functional assays using human ORL1 transfected cell membranes. On the contrary, analogues cyclized at the C-terminal parts by the disulfide linkages at positions 6-10, 7-11, 7-14, and 10-14 sustained relatively high potencies in both assays. Notably, cyclo[Cys(10), Cys(14)]NC(1-14)-NH(2) (12) was found to be a potent NC agonist nearly as active as the parent peptide or NC. The maximum efficacy (Emax) of the C-terminally cyclized analogues and their linear counterparts in the GTP gamma S functional assay showed more than 94% (vs NC as 100%), suggesting that these analogues are full agonists. Analogue 12 is the first conformationally constrained NC analogue with almost full activity, and thus may serve to analyze the bioactive conformations of NC at the receptor site as well as serving as a template for more potent NC agonists.
authors.affiliation: Tohoku Pharmaceutical University, 4-1, Komatsushima 4-chome, Aoba-ku, Sendai 981-8558, Japan.
authors.forename: A
authors.initials: A
authors.lastname: Ambo
authors.forename: N
authors.initials: N
authors.lastname: Hamazaki
authors.forename: Y
authors.initials: Y
authors.lastname: Yamada
authors.forename: E
authors.initials: E
authors.lastname: Nakata
authors.forename: Y
authors.initials: Y
authors.lastname: Sasaki
mesh_terms: D002460:Cell Line; D004220:Disulfides / Q000138:chemical synthesis* / Q000737:chemistry / Q000378:metabolism; D016244:Guanosine 5'-O-(3-Thiotriphosphate) / Q000378:metabolism; D006801:Humans; D008024:Ligands; D008968:Molecular Conformation; D018847:Opioid Peptides / Q000138:chemical synthesis* / Q000737:chemistry* / Q000378:metabolism; D009904:Optical Rotation; D010446:Peptide Fragments / Q000138:chemical synthesis* / Q000737:chemistry / Q000378:metabolism; D010456:Peptides, Cyclic / Q000138:chemical synthesis* / Q000737:chemistry / Q000378:metabolism; D011869:Radioligand Assay; D011957:Receptors, Opioid / Q000819:agonists* / Q000378:metabolism; D013329:Structure-Activity Relationship; D000094942:Nociceptin Receptor; D000097629:Nociceptin
chemical_list: D004220:Disulfides; D016244:Guanosine 5'-O-(3-Thiotriphosphate); D008024:Ligands; D018847:Opioid Peptides; D010446:Peptide Fragments; D010456:Peptides, Cyclic; D011957:Receptors, Opioid; D000094942:Nociceptin Receptor; D000097629:Nociceptin; C439969:cyclo(Cys(10,14))nociceptin (1-13) amide; C439964:nociceptin (1-13) amide; C000722283:OPRL1 protein, human