Novel analogues of neuropeptide Y with a preference for the Y-1-receptor

Neuropeptide Y (NPY) is one of the most abundant neuropeptides in the mamma lian brain and acts in humans via at least three receptor subtypes: Y-1, Y- 2, and Y-5. Whereas selective agonists and antagonists are known for the Y- 2- and Y-5-receptors, the Y-1-receptor still lacks a highly selective agoni st. This work presents the first NPY-based analogues with Y-1-receptor pref erence and agonistic properties. Furthermore, the importance of specific am ino acids of NPY for binding to the Y-receptor subtypes is presented. Among st the analogues tested, [Phe7,Pro34]pNPY (where pNPY is porcine neuropepti de Y) showed the most significant Y-1-receptor preference (> 1 : 3000-fold) , with subnanomolar affinity to the Y-1-receptor, and K-i values of approxi mate to 30 nm for the Y-2- and Y-5-subtype, respectively. Variations of pos ition 6, especially [Arg6,Pro34]pNPY and variations within positions 20-23 of NPY were found to result in further analogues with significant Y-1-recep tor preference (1 : 400-1 : 2000). In contrast, cyclo S-S [Cys20,Cys24]pNPY was found to be a highly selective ligand at the Y-2-receptor, binding onl y threefold less efficiently than NPY. Analogues containing variations of p ositions 31 and 32 showed highly reduced affinity to the Y-1-receptor, whil e binding to the Y-5-receptor was affected less. Inhibition of cAMP-accumul ation of selected peptides with replacements within position 20-23 of NPY s howed preserved agonistic properties. The NPY analogues tested give insight s into ligand-receptor interaction of NPY at the Y-1-, Y-2- and Y-5-recepto r and contribute to our understanding of subtype selectivity. Furthermore, the Y-1-receptor-preferring peptides are novel tools that will provide insi ght into the physiological role of the Y-1-receptor.