Synthesis and Pharmacological Characterization of Novel Glucagon-like Peptide-2 (GLP-2) Analogues with Low Systemic Clearance

Abstract

Glucagon-like peptide-2 receptor agonists have therapeutic potential for the treatment of intestinal diseases. The native hGLP-2, a 33 amino acid gastrointestinal peptide, is not a suitable clinical candidate, due to its very short half-life in humans. In search of GLP-2 receptor agonists with better pharmacokinetic characteristics, a series of GLP-2 analogues containing Gly substitution at position 2, norleucine in position 10, and hydrophobic substitutions in positions 11 and/or 16 was designed and synthesized. In vitro receptor potency at the human GLP-2, selectivity vs the human GLP-1 and GCG receptors, and PK profile in rats were determined for the new analogues. A number of compounds more potent at the hGLP-2R than the native hormone, showing excellent receptor selectivity and very low systemic clearance (CL) were discovered. Analogues 69 ([Gly(2),Nle(10),D-Thi(11),Phe(16)]hGLP-2-(1-30)-NH2), 72 ([Gly(2),Nle(10),D-Phe(11),Leu(16)]hGLP-2-(1-33)-OH), 73 ([Gly(2),Nle(10),D-Phe(11),Leu(16)]hGLP-2-(1-33)-NH2), 81 ([Gly(2),Nle(10),D-Phe(11),Leu(16)]hGLP-2-(1-33)-NHEt), and 85 ([Gly(2),Nle(10),D-Phe(11),Leu(16)]hGLP-2-(1-33)-NH-((CH2)2O)4-(CH2)2-CONH2) displayed the desired profiles (EC50 (hGLP-2R) < 100 pM, CL in rat <0.3 mL/min/kg, selective vs hGLP-1R and hGCGR). Compound 73 (FE 203799) was selected as a candidate for clinical development.

Pharmacological Characterization of Apraglutide, a Novel Long-Acting Peptidic Glucagon-Like Peptide-2 Agonist, for the Treatment of Short Bowel Syndrome

“…Apraglutide is a novel GLP-2 analogue with very low clearance. Its few amino acid substitutions (Table 1; Wiśniewski et al, 2011;Wiśniewski et al, 2016) confer unique PK properties, namely very low clearance and high protein binding, that enable a long in vivo halflife without conjugation or other major modifications to the peptide. The direct comparisons reported here for apraglutide versus native hGLP-2 and other GLP-2 analogues currently in the clinic (teduglutide) or in clinical trials (glepaglutide) for the treatment of SBS show that apraglutide has superior PK and pharmacodynamic profiles.…”

“…Peptide Synthesis. Peptides used in these studies were prepared as trifluoroacetic acid (TFA) salts by solid-phase peptide synthesis and purified by reverse-phase high performance liquid chromatography (HPLC) as previously described (Wiśniewski, et al 2011;Wiśniewski, et al 2016,). The peptide identities were verified by mass spectrometry.…”

“…The GLP-2 analogue apraglutide ([Gly 2 , Nle 10 , D-Phe 11 , Leu 16 ] hGLP-2 (1-33)-NH2), identified through chemistry structure-activity relationship (SAR) studies of lipophilic amino acid substitutions in positions 11 and 16 of [Gly 2 ] hGLP-2 (1-33), showed exceptionally low clearance in in vivo rat pharmacokinetic (PK) screening (Wiśniewski et al, 2011;Wiśniewski et al, 2016). This paper presents the pharmacology and PK of apraglutide compared directly with teduglutide and glepaglutide.…”

Site-Specific and Temporal Effects of Apraglutide, a Novel Long-Acting Glucagon-Like Peptide-2 Receptor Agonist, on Intestinal Growth in Mice

“…In the present pre-clinical study, we have examined the trophic effects of a novel long-acting human GLP-2R agonist, apraglutide (Wisniewski, 2016), on small intestinal and colonic weight and length, as well as small intestinal crypt-villus height and crypt number in mice, directly comparing the temporal responses to chronic treatment for 3, 7 and 10 weeks. Apraglutide is a new GLP-2 analogue which distinguish from other GLP-2 analogs such as teduglutide (Revestive ® /Gattex ® , Shire Inc.) due to its very low clearance, long elimination half-life (30 hours) and high plasma protein binding ((Hargrove, 2020) and data not shown).…”

Stereochemical inversion as a route to improved biophysical properties of therapeutic peptides exemplified by glucagon

“…A fair number of peptides of increased potency through modification with D-residues have also been reported 7,[9][10][11][12][13][14] . More recently, a systematic D-amino acid scan was used to identify GLP-2 (glucagon-like peptide-2) analogs with enhanced pharmacokinetics 15 . Consequently, a D-amino acid scan 16,17 conducted alongside the more common alanine scan 18,19 can serve to more fully interrogate the relationship between structure and function in novel peptides and proteins 20,21 .…”