Lærke S Gasbjerg1,2,3, Mikkel B Christensen1,4,5, Bolette Hartmann2,3, Amalie R Lanng1, Alexander H Sparre-Ulrich2,3, Maria B N Gabe2,3, Flemming Dela2,6, Tina Vilsbøll1,4,7, Jens J Holst2,3, Mette M Rosenkilde2,3, Filip K Knop8,9,10. 1. Center for Diabetes Research, Gentofte Hospital, University of Copenhagen, Kildegårdsvej 28, 2900, Hellerup, Denmark. 2. Department of Biomedical Sciences, Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen, Denmark. 3. Novo Nordisk Foundation Center for Basic Metabolic Research, Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen, Denmark. 4. Department of Clinical Medicine, Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen, Denmark. 5. Department of Clinical Pharmacology, Bispebjerg Hospital, University of Copenhagen, Copenhagen, Denmark. 6. Department of Geriatrics, Bispebjerg Hospital, University of Copenhagen, Copenhagen, Denmark. 7. Steno Diabetes Center Copenhagen, University of Copenhagen, Gentofte, Denmark. 8. Center for Diabetes Research, Gentofte Hospital, University of Copenhagen, Kildegårdsvej 28, 2900, Hellerup, Denmark. filipknop@dadlnet.dk. 9. Novo Nordisk Foundation Center for Basic Metabolic Research, Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen, Denmark. filipknop@dadlnet.dk. 10. Department of Clinical Medicine, Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen, Denmark. filipknop@dadlnet.dk.
Abstract
AIMS/HYPOTHESIS: Glucose-dependent insulinotropic polypeptide (GIP) is an incretin hormone secreted postprandially from enteroendocrine K cells, but despite therapeutically interesting effects, GIP physiology in humans remains incompletely understood. Progress in this field could be facilitated by a suitable GIP receptor antagonist. For the first time in humans, we investigated the antagonistic properties of the naturally occurring GIP(3-30)NH2 in in vivo and in in vitro receptor studies. METHODS: In transiently transfected COS-7 cells, GIP(3-30)NH2 was evaluated with homologous receptor binding and receptor activation (cAMP accumulation) studies at the glucagon-like peptide 1 (GLP-1), glucagon-like peptide-2 (GLP-2), glucagon, secretin and growth hormone-releasing hormone (GHRH) receptors. Ten healthy men (eligibility criteria: age 20-30 years, HbA1c less than 6.5% [48 mmol/mol] and fasting plasma glucose [FPG] less than 7 mmol/l) were included in the clinical study. Data were collected as plasma and serum samples from a cubital vein cannula. As primary outcome, insulin secretion and glucose requirements were evaluated together with in a randomised, four-period, crossover design by infusing GIP(3-30)NH2 (800 pmol kg-1 min-1), GIP (1.5 pmol kg-1 min-1), a combination of these or placebo during hyperglycaemic clamp experiments. The content of the infusions were blinded to the study participants and experimental personnel. No study participants dropped out. RESULTS:GIP(3-30)NH2 neither bound, stimulated nor antagonised a series of related receptors in vitro. The elimination plasma half-life of GIP(3-30)NH2 in humans was 7.6 ± 1.4 min. Markedly larger amounts of glucose were required to maintain the clamp during GIP infusion compared with the other days. GIP-induced insulin secretion was reduced by 82% (p < 0.0001) during co-infusion with GIP(3-30)NH2, and the need for glucose was reduced to placebo levels. There were no effects of GIP(3-30)NH2 alone or of GIP with or without GIP(3-30)NH2 on plasma glucagon, GLP-1, somatostatin, triacylglycerols, cholesterol, glycerol or NEFA. GIP(3-30)NH2 administration was well tolerated and without side effects. CONCLUSIONS/ INTERPRETATION: We conclude that GIP(3-30)NH2 is an efficacious and specific GIP receptor antagonist in humans suitable for studies of GIP physiology and pathophysiology. TRIAL REGISTRATION: ClinicalTrials.gov registration no. NCT02747472. FUNDING: The study was funded by Gangstedfonden, the European Foundation for the Study of Diabetes, and Aase og Ejnar Danielsens fond.
RCT Entities:
AIMS/HYPOTHESIS: Glucose-dependent insulinotropic polypeptide (GIP) is an incretin hormone secreted postprandially from enteroendocrine K cells, but despite therapeutically interesting effects, GIP physiology in humans remains incompletely understood. Progress in this field could be facilitated by a suitable GIP receptor antagonist. For the first time in humans, we investigated the antagonistic properties of the naturally occurring GIP(3-30)NH2 in in vivo and in in vitro receptor studies. METHODS: In transiently transfected COS-7 cells, GIP(3-30)NH2 was evaluated with homologous receptor binding and receptor activation (cAMP accumulation) studies at the glucagon-like peptide 1 (GLP-1), glucagon-like peptide-2 (GLP-2), glucagon, secretin and growth hormone-releasing hormone (GHRH) receptors. Ten healthy men (eligibility criteria: age 20-30 years, HbA1c less than 6.5% [48 mmol/mol] and fasting plasma glucose [FPG] less than 7 mmol/l) were included in the clinical study. Data were collected as plasma and serum samples from a cubital vein cannula. As primary outcome, insulin secretion and glucose requirements were evaluated together with in a randomised, four-period, crossover design by infusing GIP(3-30)NH2 (800 pmol kg-1 min-1), GIP (1.5 pmol kg-1 min-1), a combination of these or placebo during hyperglycaemic clamp experiments. The content of the infusions were blinded to the study participants and experimental personnel. No study participants dropped out. RESULTS:GIP(3-30)NH2 neither bound, stimulated nor antagonised a series of related receptors in vitro. The elimination plasma half-life of GIP(3-30)NH2 in humans was 7.6 ± 1.4 min. Markedly larger amounts of glucose were required to maintain the clamp during GIP infusion compared with the other days. GIP-induced insulin secretion was reduced by 82% (p < 0.0001) during co-infusion with GIP(3-30)NH2, and the need for glucose was reduced to placebo levels. There were no effects of GIP(3-30)NH2 alone or of GIP with or without GIP(3-30)NH2 on plasma glucagon, GLP-1, somatostatin, triacylglycerols, cholesterol, glycerol or NEFA. GIP(3-30)NH2 administration was well tolerated and without side effects. CONCLUSIONS/ INTERPRETATION: We conclude that GIP(3-30)NH2 is an efficacious and specific GIP receptor antagonist in humans suitable for studies of GIP physiology and pathophysiology. TRIAL REGISTRATION: ClinicalTrials.gov registration no. NCT02747472. FUNDING: The study was funded by Gangstedfonden, the European Foundation for the Study of Diabetes, and Aase og Ejnar Danielsens fond.
Entities:
Keywords:
Class B G protein-coupled receptor (GPCR); Glucose-dependent insulinotropic polypeptide (GIP); Hyperglycaemic clamp; Incretin physiology; Insulin secretion in vivo; Pharmacology
Authors: Lisa Getty-Kaushik; Diane H Song; Michael O Boylan; Barbara E Corkey; M Michael Wolfe Journal: Obesity (Silver Spring) Date: 2006-07 Impact factor: 5.002
Authors: Victor A Gault; Finbarr P M O'Harte; Patrick Harriott; Peter R Flatt Journal: Biochem Biophys Res Commun Date: 2002-02-08 Impact factor: 3.575
Authors: J Schirra; M Nicolaus; H J Woerle; C Struckmeier; M Katschinski; B Göke Journal: Neurogastroenterol Motil Date: 2009-02-06 Impact factor: 3.598
Authors: Piotr A Mroz; Brian Finan; Vasily Gelfanov; Bin Yang; Matthias H Tschöp; Richard D DiMarchi; Diego Perez-Tilve Journal: Mol Metab Date: 2018-12-05 Impact factor: 7.422
Authors: Shu-Chen Lu; Michelle Chen; Larissa Atangan; Elizabeth A Killion; Renee Komorowski; Yuan Cheng; Chawita Netirojjanakul; James R Falsey; Marina Stolina; Denise Dwyer; Clarence Hale; Shanaka Stanislaus; Todd Hager; Veena A Thomas; John M Harrold; David J Lloyd; Murielle M Véniant Journal: Cell Rep Med Date: 2021-04-30
Authors: Kleopatra Alexiadou; Joyceline Cuenco; James Howard; Nicolai Jacob Wewer Albrechtsen; Ibiyemi Ilesanmi; Anna Kamocka; George Tharakan; Preeshila Behary; Paul R Bech; Ahmed R Ahmed; Sanjay Purkayastha; Robert Wheller; Matthieu Fleuret; Jens Juul Holst; Stephen R Bloom; Bernard Khoo; Tricia M-M Tan Journal: BMJ Open Diabetes Res Care Date: 2020-03