title: Structure-activity relationships with neuropeptide Y analogues: a comparison of human Y1-, Y2- and rat Y2-like systems.
abstract: A structure-activity study utilising 36 synthetic Ala-analogues of the 36-residue oligopeptide neuropeptide Y (NPY) has been performed with mucosal preparations from the rat jejunum (Y2-like receptor) and compared with receptor displacement binding in the human neuroblastoma cell lines, SMS-KAN, (Y2-receptors) and SK-N-MC cells (Y1-receptors). Each amino acid of the natural sequence was replaced by L-alanine, and the four intrinsic alanine residues at position 12, 14, 18 and 23 were replaced by glycine. The purified peptides were characterized by electrospray mass spectrometry, analytical HPLC and amino acid analysis. Binding was investigated using membranes prepared from either SMS-KAN or SK-N-MC cells. The activity of each Ala-NPY analogue was assessed in mucosal preparations of rat jejunum, where NPY and PYY exert antisecretory responses which are Y2-like in pharmacology. Fourteen analogues with L-alanine replacements at position 3, 5, 8, 13, 20, 21, 22, 26, 27, 28, 29, 30, 34 and 36 were selected, none of which exhibited any antagonism of NPY responses. An order of agonist potency showed [Ala3] NPY and [Ala30] NPY equipotent with NPY, a 4-20-fold loss of activity with [Ala5] NPY, [Ala13] NPY, [Ala20] NPY, [Ala21] NPY and [Ala22] NPY; a 50-100-fold loss of activity, [Ala8] NPY, [Ala27] NPY, [Ala28] NPY and [Ala36] NPY, while [Ala34] NPY was inactive. This structure-activity relationship is similar to, but not the same as that observed in Y2-expressing SMS-KAN cells.
authors.affiliation: Department of Pharmacology, UMDS, St. Thomas' Medical School, London, UK. h.cox@umds.ac.uk
authors.forename: H M
authors.initials: HM
authors.lastname: Cox
authors.forename: I R
authors.initials: IR
authors.lastname: Tough
authors.forename: N
authors.initials: N
authors.lastname: Ingenhoven
authors.forename: A G
authors.initials: AG
authors.lastname: Beck-Sickinger
mesh_terms: D000595:Amino Acid Sequence; D019943:Amino Acid Substitution; D000818:Animals; D002460:Cell Line; D006801:Humans; D066298:In Vitro Techniques; D007413:Intestinal Mucosa / Q000187:drug effects / Q000378:metabolism; D007583:Jejunum / Q000187:drug effects / Q000378:metabolism; D008297:Male; D008969:Molecular Sequence Data; D009478:Neuropeptide Y / Q000031:analogs & derivatives* / Q000737:chemistry / Q000494:pharmacology; D051381:Rats; D017207:Rats, Sprague-Dawley; D017476:Receptors, Neuropeptide Y / Q000187:drug effects / Q000378:metabolism; D013329:Structure-Activity Relationship
chemical_list: D009478:Neuropeptide Y; D017476:Receptors, Neuropeptide Y; C097212:neuropeptide Y-Y1 receptor; C095889:neuropeptide Y2 receptor