Exendin-4的固相化学合成及鉴定

中国生物工程杂志

2011, Vol. 31

Issue (02): 69-73

技术与方法

Exendin-4的固相化学合成及鉴定

刘延杰, 季虹, 林鲁霞, 臧学章, 宋长征, 荣海钦

山东省内分泌与代谢病研究所山东省医学科学院济南 250062

Solid Phase Peptide Synthesis and Analysis for Exendin-4

LIU Yan-jie, JI Hong, LIN Lu-xia, ZANG Xue-zhang, SONG Chang-zheng, RONG Hai-qin

Shandong Institute of Endocrine and Metabolic Diseases, Shandong Academy of Medical Science, Jinan 250062, China

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摘要：

Exendin-4是首个获准上市的肠促胰素类似物药物,可模拟人体自身激素GLP-1的功能,不仅改善血糖,还能促进胰岛β细胞新生、增殖,抑制β细胞凋亡,改善β细胞功能,促进胰岛素分泌,增加机体对胰岛素的敏感性。根据Exendin-4的潜在市场价值,以Fmoc固相合成策略为基础,NMP为反应溶剂,以HOBt/DIC为缩合剂,添加盖帽程序,优化合成工艺条件。肽树脂裂解采用TFA/Phenol/TIS裂解液,并应用高效液相色谱和四级杆-飞行时间串联质谱对其进行分析鉴定和纯化,最终获得Exendin-4,产率为21%,纯度为99.4%。活性测定结果显示,合成的Exendin-4显著提高胰岛瘤细胞的活性,并呈一定剂量依赖性。

关键词： Exendin-4; 固相化学合成; 2型糖尿病

Abstract:

Exendin-4 is the first-approved drug of the incretin mimetic family, which acts like the naturally occurring hormone GLP-1 in vivo. Exendin-4 decreases fasting and postprandial plasma glucose levels. Exendin-4 may play a role in enhancing glucose-dependent stimulation of insulin secretion, and maintaining of β-cell mass by promotion of β-cell proliferation and neogenesis, and inhibiting of β-cell apoptosis. This experiment studied the standard Fmoc strategy solid phase synthesis of Exendin-4. Using NMP as the coupling solvent, HOBt/DIC as the coupling reagent, AIM as the containing reagent. Synthesized peptides were clearaged by TFA/Phenol/TIS, and identified by high performance liquid chromatography and quadrupole time of flight LC/MS. The yield of pure Exendin-4 was 21% with a purity of 99.4%. Using the SRB method to measure the bioassay of Exendin-4 cell regeneration experiment. It is an influential research into the therapy of type 2 diabetes mellitus.

Key words: Exendin-4 Solid phase peptide synthesis Type 2 diabetes mellitus

收稿日期: 2010-11-02 出版日期: 2011-02-18

ZTFLH:

Q819

基金资助:

山东省科技攻关计划(2009GG20002046、2009GG20002049)资助项目

通讯作者: 荣海钦 E-mail: hqrong@live.cn

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引用本文:

刘延杰, 季虹, 林鲁霞, 臧学章, 宋长征, 荣海钦. Exendin-4的固相化学合成及鉴定[J]. 中国生物工程杂志, 2011, 31(02): 69-73.

LIU Yan-jie, JI Hong, LIN Lu-xia, ZANG Xue-zhang, SONG Chang-zheng, RONG Hai-qin. Solid Phase Peptide Synthesis and Analysis for Exendin-4. China Biotechnology, 2011, 31(02): 69-73.

链接本文:

https://manu60.magtech.com.cn/biotech/CN/ 或 https://manu60.magtech.com.cn/biotech/CN/Y2011/V31/I02/69

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