The effect of fatty diacid acylation of human PYY3-36 on Y2 receptor potency and half-life in minipigs

Abstract

Peptides are notoriously known to display very short in vivo half-lives often measured in minutes which in many cases greatly reduces or eliminates sufficient in vivo efficacy. To obtain long half-lives allowing for up to once-weekly dosing regimen, fatty acid acylation (lipidation) have been used to non-covalently associate the peptide to serum albumin thus serving as a circulating depot. This approach is generally considered in the scientific and patent community as a standard approach to protract almost any given peptide. However, it is not trivial to prolong the half-life of peptides by lipidation and still maintain high potency and good formulation properties. Here we show that attaching a fatty acid to the obesity-drug relevant peptide PYY3-36 is not sufficient for long pharmacokinetics (PK), since the position in the backbone, but also type of fatty acid and linker strongly influences PK and potency. Furthermore, understanding the proteolytic stability of the backbone is key to obtain long half-lives by lipidation, since backbone cleavage still occurs while associated to albumin. Having identified a PYY analogue with a sufficient half-life, we show that in combination with a GLP-1 analogue, liraglutide, additional weight loss can be achieved in the obese minipig model.

BI 456906: Discovery and preclinical pharmacology of a novel GCGR/GLP-1R dual agonist with robust anti-obesity efficacy

“…Prolonging the in vivo half-life of peptides by acylation to promote high-affinity albumin binding is a well-established approach to maximizing the pharmacological potential of peptide hormone mimetics [ 52 ]. However, the complexity of peptide lipidation with variations in the lengths of fatty acids, linkers, sites of attachment, and physicochemical properties has been acknowledged in recent years [ 34 , [53] , [54] , [55] , [56] ]. Specifically, optimization of these parameters is relevant to their affinities to the target receptors, plasma proteins, and plasma membranes [ 53 , 55 , 57 ].…”

“…However, the complexity of peptide lipidation with variations in the lengths of fatty acids, linkers, sites of attachment, and physicochemical properties has been acknowledged in recent years [ 34 , [53] , [54] , [55] , [56] ]. Specifically, optimization of these parameters is relevant to their affinities to the target receptors, plasma proteins, and plasma membranes [ 53 , 55 , 57 ]. All these changes, together with amino acid sequence modifications, influenced the pharmacological profile of the peptide at the target receptors and it was thus important to establish that an appropriate profile had been obtained.…”

Tirzepatide, a New Era of Dual-Targeted Treatment for Diabetes and Obesity: A Mini-Review

“…More specifically, the structure is based on the native GIP sequence and includes C20 fatty diacid moiety (eicosanedioic acid) linked via hydrophilic linkers (γ-Glu-2xAdo, gamma glutamate and bis-aminodiethoxyacetyl) connected to lysine residue at C20 position [31]. The peptide sequence of tirzepatide contains two non-coded amino acid residues (Aib, α-amino isobutyric acid) at position 2 and 13, which are responsible for its long half-life and high affinity to albumin [32]. The C-terminus of the peptide is amidated (Figure 2) [33].…”

Basal insulin intensification with GLP-1RA and dual GIP and GLP-1RA in patients with uncontrolled type 2 diabetes mellitus: A rapid review of randomized controlled trials and meta-analysis

“…A 20-carbon fatty acid linked to the lysine residue in position 20 of the primary structure allows TZP to be bound to circulating albumin. Acylation technology, previously adopted and developed for Liraglutide and Semaglutide, increases TZP half-life to around five days, making it administrable once weekly (28,(31)(32)(33).…”