A Mixed Mirror-image DNA/RNA Aptamer Inhibits Glucagon and Acutely Improves Glucose Tolerance in Models of Type 1 and Type 2 Diabetes

被引：65

作者：

Vater, Axel ^{[1
]}

Sell, Simone ^{[1
]}

Kaczmarek, Przemyslaw ^{[2
]}

Maasch, Christian ^{[1
]}

Buchner, Klaus ^{[1
]}

Pruszynska-Oszmalek, Ewa ^{[2
]}

Kolodziejski, Pawel ^{[2
]}

Purschke, Werner G. ^{[1
]}

Nowak, Krzysztof W. ^{[2
]}

Strowski, Mathias Z. ^{[3
,4
]}

Klussmann, Sven ^{[1
]}

机构：

[1] NOXXON Pharma AG, D-10589 Berlin, Germany

[2] Poznan Univ Life Sci, Dept Anim Physiol & Biochem, PL-60637 Poznan, Poland

[3] Charite, Dept Gastroenterol & Hepatol, D-13353 Berlin, Germany

[4] Charite, Interdisciplinary Ctr Metab Endocrinol Diabet & M, D-13353 Berlin, Germany

来源：

JOURNAL OF BIOLOGICAL CHEMISTRY | 2013年 / 288卷 / 29期

关键词：

ENDOGENOUS GLUCAGON; RANDOM MUTAGENESIS; MOUSE MODELS; IN-VIVO; INSULIN; HYPERGLYCEMIA; SPIEGELMER; RNA; IMMUNONEUTRALIZATION; FLEXIBILITY;

D O I：

10.1074/jbc.M112.444414

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

070307 [化学生物学]; 071010 [生物化学与分子生物学];

摘要：

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 (R)) 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 K-d 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.

引用

页码：21136 / 21147

页数：12

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