title: Enzymatic mono-pegylation of glucagon-like peptide 1 towards long lasting treatment of type 2 diabetes.
abstract: Human glucagon-like peptide-1 (GLP-1) is a physiological gastrointestinal peptide with glucose-dependent insulinotropic effects which is therefore considered an interesting antidiabetic agent. However, after in vivo administration, exogenous GLP-1 does not exert its physiological action due to the combination of rapid proteolytic degradation by ubiquitous dipeptidyldipeptidase IV (DPP IV) enzyme and renal clearance resulting in an extremely short circulating half-life. In this work we describe the conjugation of GLP-1-(7-36)-amide derivatives with polyethylene glycol (PEG) by enzymatic site-specific transglutamination reaction as an approach to reduce both the proteolysis and the renal clearance rates. The compound GLP-1-(7-36)-amide-Q(23)-PEG 20 kDa monopegylated on the single glutamine residue naturally present in position 23 maintained the ability to activate the GLP-1 receptor expressed in the rat β-cell line RIN-m5F with nanomolar potency along with an increased in vitro resistance to DDP IV and a circulating half-life of about 12 h after subcutaneous administration in rats. These properties enabled GLP-(7-36)-amide-Q(23)-PEG 20 kDa to exert a glucose-stabilizing effect for a period as long as 8 h, as demonstrated by a single subcutaneous injection to diabetic mice concomitantly challenged with an oral glucose load. The results reported in this work indicate that GLP-(7-36)-amide-Q(23)-PEG 20 kDa could be a lead compound for the development of long-lasting anti-diabetic agents useful in the treatment of type 2 diabetes affected patients.
authors.affiliation: Bioker srl-Multimedica Group, c/o CNR-IGB, via P. Castellino 111, 80131 Naples, Italy.
authors.forename: Fabio
authors.initials: F
authors.lastname: Selis
authors.affiliation: Bioker srl-Multimedica Group, c/o Sardinia Scientific and Technological Park, Building 3, I-09010 Pula (Cagliari), Italy.
authors.forename: Rodolfo
authors.initials: R
authors.lastname: Schrepfer
authors.affiliation: Bioker srl-Multimedica Group, c/o Department of Neuroscience, via Pansini 5, 80131 Naples, Italy.
authors.forename: Riccardo
authors.initials: R
authors.lastname: Sanna
authors.affiliation: Bioker srl-Multimedica Group, c/o CNR-IGB, via P. Castellino 111, 80131 Naples, Italy.
authors.forename: Silvia
authors.initials: S
authors.lastname: Scaramuzza
authors.affiliation: Bioker srl-Multimedica Group, c/o Sardinia Scientific and Technological Park, Building 3, I-09010 Pula (Cagliari), Italy.
authors.forename: Giancarlo
authors.initials: G
authors.lastname: Tonon
authors.affiliation: Section of Biochemical Pharmacology, Department of Neuroscience, University of Cagliari, 09042 Monserrato (Cagliari), Italy.
authors.forename: Simona
authors.initials: S
authors.lastname: Dedoni
authors.affiliation: Section of Biochemical Pharmacology, Department of Neuroscience, University of Cagliari, 09042 Monserrato (Cagliari), Italy.
authors.forename: Pierluigi
authors.initials: P
authors.lastname: Onali
authors.affiliation: Bioker srl-Multimedica Group, c/o Sardinia Scientific and Technological Park, Building 3, I-09010 Pula (Cagliari), Italy.
authors.forename: Gaetano
authors.initials: G
authors.lastname: Orsini
authors.affiliation: Diabetes and Metabolic Diseases Unit, IRCCS Multimedica, Sesto San Giovanni (Milano), Italy.
authors.forename: Stefano
authors.initials: S
authors.lastname: Genovese
keywords: Enzymatic pegylation; GLP-1; Pegylated GLP-1; Transglutaminase; Type 2 diabetes