title: A mixed mirror-image DNA/RNA aptamer inhibits glucagon and acutely improves glucose tolerance in models of type 1 and type 2 diabetes.
abstract: Excessive secretion of glucagon, a functional insulin antagonist, significantly contributes to hyperglycemia in type 1 and type 2 diabetes. Accordingly, immunoneutralization of glucagon or genetic deletion of the glucagon receptor improved glucose homeostasis in animal models of diabetes. Despite this strong evidence, agents that selectively interfere with endogenous glucagon have not been implemented in clinical practice yet. We report the discovery of mirror-image DNA-aptamers (Spiegelmer®) that bind and inhibit glucagon. The affinity of the best binding DNA oligonucleotide was remarkably increased (>25-fold) by the introduction of oxygen atoms at selected 2'-positions through deoxyribo- to ribonucleotide exchanges resulting in a mixed DNA/RNA-Spiegelmer (NOX-G15) that binds glucagon with a Kd of 3 nm. NOX-G15 shows no cross-reactivity with related peptides such as glucagon-like peptide-1, glucagon-like peptide-2, gastric-inhibitory peptide, and prepro-vasoactive intestinal peptide. In vitro, NOX-G15 inhibits glucagon-stimulated cAMP production in CHO cells overexpressing the human glucagon receptor with an IC50 of 3.4 nm. A single injection of NOX-G15 ameliorated glucose excursions in intraperitoneal glucose tolerance tests in mice with streptozotocin-induced (type 1) diabetes and in a non-genetic mouse model of type 2 diabetes. In conclusion, the data suggest NOX-G15 as a therapeutic candidate with the potential to acutely attenuate hyperglycemia in type 1 and type 2 diabetes.
authors.affiliation: From the NOXXON Pharma AG, Max-Dohrn-Strasse 8-10, 10589 Berlin, Germany.
authors.forename: Axel
authors.initials: A
authors.lastname: Vater
authors.affiliation: From the NOXXON Pharma AG, Max-Dohrn-Strasse 8-10, 10589 Berlin, Germany.
authors.forename: Simone
authors.initials: S
authors.lastname: Sell
authors.affiliation: the Department of Animal Physiology and Biochemistry, Poznan University of Life Sciences, 35 Wolynska Street, 60637 Poznan, Poland, and.
authors.forename: Przemyslaw
authors.initials: P
authors.lastname: Kaczmarek
authors.affiliation: From the NOXXON Pharma AG, Max-Dohrn-Strasse 8-10, 10589 Berlin, Germany.
authors.forename: Christian
authors.initials: C
authors.lastname: Maasch
authors.affiliation: From the NOXXON Pharma AG, Max-Dohrn-Strasse 8-10, 10589 Berlin, Germany.
authors.forename: Klaus
authors.initials: K
authors.lastname: Buchner
authors.affiliation: the Department of Animal Physiology and Biochemistry, Poznan University of Life Sciences, 35 Wolynska Street, 60637 Poznan, Poland, and.
authors.forename: Ewa
authors.initials: E
authors.lastname: Pruszynska-Oszmalek
authors.affiliation: the Department of Animal Physiology and Biochemistry, Poznan University of Life Sciences, 35 Wolynska Street, 60637 Poznan, Poland, and.
authors.forename: Pawel
authors.initials: P
authors.lastname: Kolodziejski
authors.affiliation: From the NOXXON Pharma AG, Max-Dohrn-Strasse 8-10, 10589 Berlin, Germany.
authors.forename: Werner G
authors.initials: WG
authors.lastname: Purschke
authors.affiliation: the Department of Animal Physiology and Biochemistry, Poznan University of Life Sciences, 35 Wolynska Street, 60637 Poznan, Poland, and.
authors.forename: Krzysztof W
authors.initials: KW
authors.lastname: Nowak
authors.affiliation: the Department of Hepatology and Gastroenterology and Interdisciplinary Centre of Metabolism: Endocrinology, Diabetes and Metabolism, Charité-Universitätsmedizin Berlin, Augustenburger Platz 1, 13353 Berlin, Germany.
authors.forename: Mathias Z
authors.initials: MZ
authors.lastname: Strowski
authors.affiliation: From the NOXXON Pharma AG, Max-Dohrn-Strasse 8-10, 10589 Berlin, Germany,. Electronic address: sklussmann@noxxon.com.
authors.forename: Sven
authors.initials: S
authors.lastname: Klussmann
mesh_terms: D000818:Animals; D052157:Aptamers, Nucleotide / Q000097:blood / Q000493:pharmacokinetics / Q000494:pharmacology* / Q000627:therapeutic use; D001786:Blood Glucose / Q000378:metabolism; D001835:Body Weight / Q000187:drug effects; D016466:CHO Cells; D006224:Cricetinae; D003412:Cricetulus; D000242:Cyclic AMP / Q000096:biosynthesis; D003922:Diabetes Mellitus, Type 1 / Q000097:blood* / Q000188:drug therapy; D003924:Diabetes Mellitus, Type 2 / Q000097:blood* / Q000188:drug therapy; D004195:Disease Models, Animal; D005215:Fasting / Q000097:blood; D005934:Glucagon / Q000037:antagonists & inhibitors* / Q000378:metabolism; D005951:Glucose Tolerance Test; D006801:Humans; D007700:Kinetics; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D012313:RNA / Q000378:metabolism*
chemical_list: D052157:Aptamers, Nucleotide; D001786:Blood Glucose; D012313:RNA; D005934:Glucagon; D000242:Cyclic AMP
keywords: Aptamers; DNA; Diabetes; Drug Design; Drug Discovery; Glucagon Inhibitor; Mirror-image Aptamer; Spiegelmer; Type 1 Diabetes; Type 2 Diabetes