Design of Y₂ Receptor Selective and Proteolytically Stable PYY_3–36 Analogues

Abstract

In recent years peptide YY (PYY) has attracted attention within the area of diabetes and obesity due to its involvement in food intake regulation and glucose homeostasis. It is well-known that PYY1–36 is rapidly cleaved by dipeptidyl peptidase-4 to the more Y2 receptor selective analogue PYY3–36, which is further cleaved to the inactive analogue PYY3–34. In order to improve the selectivity and proteolytic stability of the C-terminus, we synthesized several analogues incorporating N-methyl amino acids or β-homo amino acids and other non-natural amino acids. These were tested against all four NPY receptors, and highly potent and Y2 receptor selective analogues were identified by combining a tryptophan residue in position 30 with either N-methyl or β-homo arginine in position 35. We also identified an analogue with a MeGln34 substitution that surprisingly displayed high affinity toward all four receptors. In addition, these analogues displayed improved stability toward C-terminal proteolysis compared to native PYY3–36.

Modified Peptide YY Molecule Attenuates the Activity of NPY/AgRP Neurons and Reduces Food Intake in Male Mice

“…This, along with its short half-life, makes native PYY 3-36 unappealing as a therapeutic. Therefore, a PYY 3-36 analog, NNC0165-1273, which has at least 650-fold affinity for Y2 over the other Y receptors, as well as having a slightly increased in vitro half-life in pig plasma was developed (9). This should increase its potential as an obesity therapeutic by targeting the anorectic pathways with greater precision.…”

“…NNC0165-1273 was identified from a series of modified PYY peptides and synthesized with the aim of increased selectivity for Y2 receptors vs the other Y receptors [peptide no. 29 in Østergaard et al (9)]. NNC0165-1273 showed a binding inhibitory constant (K i ) on the human Y 2 receptor of 2 nM and >10,000, 2500, and 1300 nM K i values on the human Y1, Y4, and Y5 receptors, respectively.…”

“…Additionally, in in vitro metabolism studies, the peptide has as a half-life of 570 minutes vs 200 minutes for PYY 3-36 as measured in pig plasma. [For further information, description of the peptide and methods, see Østergaard et al (9). ]…”

“…However, secreted PYY is rapidly converted by dipeptidyl peptidase-4, so that the major circulating hormone is the truncated form, PYY 3-36 (7). This peptide retains its high affinity for Y2, but has reduced affinity for both Y1 and Y4, and around a 10-fold reduced selectivity for Y5 (8, 9), and has multiple effects on energy balance including control of satiety and gut motility (10).…”

Impact of Substitution Registry on the Receptor‐Activation Profiles of Backbone‐Modified Glucagon‐like Peptide‐1 Analogues

“…Among these drugs are synthetic peptides containing unnatural components designed to optimize therapeutic performance . Unnatural modifications in clinical agents have so far been limited to side chains, but backbone‐based modifications have been evaluated in the laboratory . We have focused on agonists in which a subset of natural α‐amino acid residues is replaced by β‐amino acid residues.…”