DPP-IV와 NEP 24.11 모두에 저항성을 가진 글루카곤 양 펩티드-1(GLP-1) 유사체 개발

다국어 초록 (Multilingual Abstract)

BACKGROUND
Glucagon-like peptide-1(7-36)amide (GLP-1) possesses several unique and beneficial effects for the treatment of type 2 diabetes. However, rapid degradation of GLP-1 results in a short half-life in vivo hindering therapeutic development. Both dipeptidyl peptidase-IV (DPP-IV) and neutral endopeptidase 24.11 (NEP 24.11) degrade GLP-1. While DPP-IV resistant analogs of GLP-1 are available, agonists resistant to NEP 24.11 have not been developed. The goal of this study is to develop a novel GLP-1 analog resistant to both DPP-IV and NEP 24.11.
METHOD
We designed several NEP 24.11 resistant GLP-1 analogs and Anygen Co. Ltd. (Gwangju, Korea) synthesized these analogs. A NEP 24.11-containing cell extract preparations were made by transfecting HeLa cells with pcDNA3 containing NEP 24.11 cDNA. HEK293T cells were plated into 24-well plates and cotransfected with pcDNA3 containing the GLP-1 receptor cDNA and CRE-luc reporter vector. Forty eight hours after transfection, cells were treated with GLP-1 analogs or NEP 24.11 extract-incubated GLP-1 analogs for 6 hours. Luciferase activities in cell extracts were determined by luciferase assay according to standard methods.
RESULTS
Human GLP-1 incubated with NEP 24.11 was completely degraded within 10 hours. When exendin-4 was incubated with NEP 24.11 for 10 hours at the same condition, the potency decreased by 50% of initial activity. Newly developed GLP-1 analogs elicited significant luciferase activity (Emax; hGa-E4, 28.17 vs. hGf-E4, 33.92 vs. hGLP-1, 34.15 vs. exendin-4, 25.30). Novel NEP 24.11-resistant GLP-1 analogs, hGa-E4 and hGf-E4, were more resistant to NEP 24.11 than human GLP-1 and exendin-4. Exendin-4 was also relatively resistant to NEP 24.11.
CONCLUSION
These studies indicate that Ser2 substitution and some portion of the exendin-4 sequence addition, Ser2 substitution and Val29 substitution and some portion of exendin-4 sequence addition result in both DPP-IV and NEP 24.11 resistant and biologically potent forms of GLP-1. These novel GLP-1 analogs will improve the longevity of GLP-1 like action in circulation, suggesting that these new analogs represent a potential candidate for future therapeutic development.

목차 (Table of Contents)

• I. 서론 = 1
• II. 방법 = 4
• III. 결과 = 8
• IV. 고찰 = 12
• V. 요약 = 19
• VI. 참고문헌 = 27