Neprilysin Inhibition Increases Glucagon Levels in Humans and Mice With Potential Effects on Amino Acid Metabolism

Sasha A S Kjeldsen; Lasse H Hansen; Nathalie Esser; Steve Mongovin; Marie Winther-Sørensen; Katrine D Galsgaard; Jenna E Hunt; Hannelouise Kissow; Frederik R Ceutz; Dijana Terzic; Peter D Mark; Peter Plomgaard; Jens P Goetze; Gijs H Goossens; Ellen E Blaak; Carolyn F Deacon; Mette M Rosenkilde; Sakeneh Zraika; Jens J Holst; Nicolai J Wewer Albrechtsen

doi:10.1210/jendso/bvab084

Neprilysin Inhibition Increases Glucagon Levels in Humans and Mice With Potential Effects on Amino Acid Metabolism

J Endocr Soc. 2021 May 16;5(9):bvab084. doi: 10.1210/jendso/bvab084. eCollection 2021 Sep 1.

Authors

Sasha A S Kjeldsen^{1

2}, Lasse H Hansen^{3

4}, Nathalie Esser^{5

6}, Steve Mongovin⁶, Marie Winther-Sørensen¹, Katrine D Galsgaard^{1

7}, Jenna E Hunt^{1

7}, Hannelouise Kissow^{1

7}, Frederik R Ceutz¹, Dijana Terzic³, Peter D Mark³, Peter Plomgaard^{3

8}, Jens P Goetze^{1

3}, Gijs H Goossens⁹, Ellen E Blaak⁹, Carolyn F Deacon¹, Mette M Rosenkilde^{1

7}, Sakeneh Zraika^{5

6}, Jens J Holst^{1

4}, Nicolai J Wewer Albrechtsen^{1

2

3}

Affiliations

¹ Department of Biomedical Sciences, Faculty of Health and Medical Sciences, University of Copenhagen, 2200 Copenhagen,Denmark.
² Novo Nordisk Foundation Center for Protein Research, Faculty of Health and Medical Sciences, University of Copenhagen, 2200 Copenhagen,Denmark.
³ Department of Clinical Biochemistry, Copenhagen University Hospital, Rigshospitalet, 2100 Copenhagen, Denmark.
⁴ Copenhagen Center for Glycomics, Department of Cellular and Molecular Medicine, Faculty of Health and Medical Sciences, University of Copenhagen, 2200 Copenhagen, Denmark.
⁵ Department of Medicine, Division of Metabolism, Endocrinology and Nutrition, University of Washington, Seattle, Washington 98195-6426, USA.
⁶ Veterans Affairs Puget Sound Health Care System, Seattle, Washington 98108, USA.
⁷ Novo Nordisk Foundation Center for Basic Metabolic Research, Faculty of Health and Medical Sciences, University of Copenhagen, 2200 Copenhagen, Denmark.
⁸ Department of Clinical Medicine, Faculty of Health and Medical Sciences, University of Copenhagen, 2200 Copenhagen, Denmark.
⁹ Department of Human Biology, NUTRIM School of Nutrition and Translational Research in Metabolism, Maastricht University Medical Centre+, Maastricht, the Netherlands.

Abstract

Context: Inhibitors of the protease neprilysin (NEP) are used for treating heart failure, but are also linked to improvements in metabolism. NEP may cleave proglucagon-derived peptides, including the glucose and amino acid (AA)-regulating hormone glucagon. Studies investigating NEP inhibition on glucagon metabolism are warranted.

Objective: This work aims to investigate whether NEP inhibition increases glucagon levels.

Methods: Plasma concentrations of glucagon and AAs were measured in eight healthy men during a mixed meal with and without a single dose of the NEP inhibitor/angiotensin II type 1 receptor antagonist, sacubitril/valsartan (194 mg/206 mg). Long-term effects of sacubitril/valsartan (8 weeks) were investigated in individuals with obesity (n = 7). Mass spectrometry was used to investigate NEP-induced glucagon degradation, and the derived glucagon fragments were tested pharmacologically in cells transfected with the glucagon receptor (GCGR). Genetic deletion or pharmacological inhibition of NEP with or without concomitant GCGR antagonism was tested in mice to evaluate effects on AA metabolism.

Results: In healthy men, a single dose of sacubitril/valsartan significantly increased postprandial concentrations of glucagon by 228%, concomitantly lowering concentrations of AAs including glucagonotropic AAs. Eight-week sacubitril/valsartan treatment increased fasting glucagon concentrations in individuals with obesity. NEP cleaved glucagon into 5 inactive fragments (in vitro). Pharmacological NEP inhibition protected both exogenous and endogenous glucagon in mice after an AA challenge, while NEP-deficient mice showed elevated fasting and AA-stimulated plasma concentrations of glucagon and urea compared to controls.

Conclusion: NEP cleaves glucagon, and inhibitors of NEP result in hyperglucagonemia and may increase postprandial AA catabolism without affecting glycemia.

Keywords: (sacubitril/valsartan); Entresto; metabolism; proglucagon-derived peptides.

Abstract

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