Christian Borch Jacobsen1, René Raavé2, Marie Østergaard Pedersen3, Pierre Adumeau4, Mathieu Moreau4, Ibai E Valverde4, Inga Bjørnsdottir5, Jesper Bøggild Kristensen6, Mette Finderup Grove5, Kirsten Raun5, James McGuire5, Victor Goncalves4, Sandra Heskamp2, Franck Denat4, Magnus Gustafsson7. 1. Global Research Technologies, Novo Nordisk A/S, Denmark; Isotope Chemistry, CMC Development, Novo Nordisk A/S, Denmark. Electronic address: cbjc@novonordisk.com. 2. Department of Radiology and Nuclear Medicine, Radboudumc, Radboud Institute for Molecular Life Sciences, Nijmegen, the Netherlands. 3. Global Research Technologies, Novo Nordisk A/S, Denmark. 4. Institut de Chimie Moléculaire de l'Université de Bourgogne, UMR CNRS 6302, Université Bourgogne Franche-Comté, France. 5. Global Drug Discovery, Novo Nordisk A/S, Denmark. 6. Isotope Chemistry, CMC Development, Novo Nordisk A/S, Denmark. 7. Global Research Technologies, Novo Nordisk A/S, Denmark. Electronic address: mbfg@novonordisk.com.
Abstract
INTRODUCTION: Lately, zirconium-89 has shown great promise as a radionuclide for PET applications of long circulating biomolecules. Here, the design and synthesis of protracted and long-lived GLP-1 receptor agonists conjugated to desferrioxamine and labelled with zirconium-89 is presented with the purpose of studying their in vivo distribution by PET imaging. The labelled conjugates were evaluated and compared to a non-labelled GLP-1 receptor agonist in both in vitro and in vivo assays to certify that the modification did not significantly alter the peptides' structure or function. Finally, the zirconium-89 labelled peptides were employed in PET imaging, providing visual verification of their in vivo biodistribution. METHODS: The evaluation of the radiolabelled peptides and comparison to their non-labelled parent peptide was performed by in vitro assays measuring binding and agonistic potency to the GLP-1 receptor, physicochemical studies aiming at elucidating change in peptide structure upon bioconjugation and labelling as well as an in vivo food in-take study illustrating the compounds' pharmacodynamic properties. The biodistribution of the labelled GLP-1 analogues was determined by ex vivo biodistribution and in vivo PET imaging. RESULTS: The results indicate that it is surprisingly feasible to design and synthesize a protracted, zirconium-89 labelled GLP-1 receptor agonist without losing in vitro potency or affinity as compared to a non-labelled parent peptide. Physicochemical properties as well as pharmacodynamic properties are also maintained. The biodistribution in rats shows high accumulation of radiolabelled peptide in well-perfused organs such as the liver, kidney, heart and lungs. The PET imaging study confirmed the findings from the biodistribution study with a significant high uptake in kidneys and presence of activity in liver, heart and larger blood vessels. CONCLUSIONS AND ADVANCES IN KNOWLEDGE: This initial study indicates the potential to monitor the in vivo distribution of long-circulating incretin hormones using zirconium-89 based PET.
INTRODUCTION: Lately, zirconium-89 has shown great promise as a radionuclide for PET applications of long circulating biomolecules. Here, the design and synthesis of protracted and long-lived GLP-1 receptor agonists conjugated to desferrioxamine and labelled with zirconium-89 is presented with the purpose of studying their in vivo distribution by PET imaging. The labelled conjugates were evaluated and compared to a non-labelled GLP-1 receptor agonist in both in vitro and in vivo assays to certify that the modification did not significantly alter the peptides' structure or function. Finally, the zirconium-89 labelled peptides were employed in PET imaging, providing visual verification of their in vivo biodistribution. METHODS: The evaluation of the radiolabelled peptides and comparison to their non-labelled parent peptide was performed by in vitro assays measuring binding and agonistic potency to the GLP-1 receptor, physicochemical studies aiming at elucidating change in peptide structure upon bioconjugation and labelling as well as an in vivo food in-take study illustrating the compounds' pharmacodynamic properties. The biodistribution of the labelled GLP-1 analogues was determined by ex vivo biodistribution and in vivo PET imaging. RESULTS: The results indicate that it is surprisingly feasible to design and synthesize a protracted, zirconium-89 labelled GLP-1 receptor agonist without losing in vitro potency or affinity as compared to a non-labelled parent peptide. Physicochemical properties as well as pharmacodynamic properties are also maintained. The biodistribution in rats shows high accumulation of radiolabelled peptide in well-perfused organs such as the liver, kidney, heart and lungs. The PET imaging study confirmed the findings from the biodistribution study with a significant high uptake in kidneys and presence of activity in liver, heart and larger blood vessels. CONCLUSIONS AND ADVANCES IN KNOWLEDGE: This initial study indicates the potential to monitor the in vivo distribution of long-circulating incretin hormones using zirconium-89 based PET.
Authors: Eduardo Felipe Alves Fernandes; Jonas Wilbs; Rene Raavé; Christian Borch Jacobsen; Hanne Toftelund; Hans Helleberg; Milou Boswinkel; Sandra Heskamp; Magnus Bernt Frederik Gustafsson; Inga Bjørnsdottir Journal: ACS Pharmacol Transl Sci Date: 2022-06-30
Authors: Stacey E Rudd; Jessica K Van Zuylekom; Anna Raicevic; Lesley A Pearce; Carleen Cullinane; Charlotte C Williams; Timothy E Adams; Rodney J Hicks; Paul S Donnelly Journal: Chem Sci Date: 2021-05-25 Impact factor: 9.825