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化学学报
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化学学报 ›› 2011, Vol. 69 ›› Issue (23): 2889-2895.DOI: 10.6023/A1103042 上一篇    下一篇

研究论文

胰岛素的mPEG修饰及其活性表征

吴志民1, 蒋薇1, 杨友强1, 周丽英2, 凌莉1, 徐峰1, 郭新东1, 章莉娟*,1   

  1. (1 华南理工大学化学与化工学院 广州 510640)
    (2 香港科技大学化学与生物分子工程系 香港)
  • 投稿日期:2011-03-04 修回日期:2011-06-10 发布日期:2011-08-15
  • 通讯作者: 章莉娟 E-mail:celjzh@scut.edu.cn
  • 基金资助:

    国家自然科学基金;广东省自然科学基金;广州市应用基础研究计划;香港创新科技署项目;化学工程联合国家重点实验室开放课题资助

mPEGylation of Insulin and Characterization of Its Activity

Wu Zhimin1; Jiang Wei1; Yang Youqiang1; Zhou Liying2; Ling Li1; Xu Feng1; Guo Xindong1; Zhang Lijuan*,1   

  1. (1 School of Chemistry and Chemical Engineering, South China University of Technology, Guangzhou 510640)
    (2 Department of Chemical and Biomolecular Engineering, The Hong Kong University of Science and Technology, Hong Kong)
  • Received:2011-03-04 Revised:2011-06-10 Published:2011-08-15

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用单甲氧基聚乙二醇-丙醛(mPEG-ALD)、单甲氧基聚乙二醇-丁醛(mPEG-ButyrALD)和单甲氧基聚乙二醇-琥珀酰亚胺碳酸酯(mPEG-SC)对猪胰岛素进行化学修饰. 考察了反应时间、pH、物质的量比及分子量等因素对修饰效果的影响. 通过HPLC, MALDI-TOF-MS(基质辅助激光解吸时间飞行质谱), SDS-PAGE(十二烷基硫酸钠-聚丙烯酰胺凝胶电泳)和TNBS(三硝基苯磺酸钠法)等表征发现mPEG-ButyrALD具有更高的修饰选择性和修饰率. 圆二色光谱(CD)分析修饰产物的二级结构发现, 胰岛素经mPEG衍生物修饰后, 二级结构基本没有改变. 动物实验表明, mPEG-ButyrALD修饰胰岛素后半衰期延长, 因而可以延长降血糖作用时间, 改善胰岛素药效.

关键词: 胰岛素, 聚乙二醇衍生物, 修饰, 二级结构, 降血糖效果

Pork insulin was modified by three kinds of mPEG derivatives (mPEG-ALD, mPEG-ButyrALD and mPEG-SC) with different active moieties. The effects of reaction time, pH, molar ratio and molecular weight on the modification rate and site-selectivity were investigated. The results showed that mPEG-ButyrALD-INS had higher site-selectivity and modification rate compared to the other two modifiers characterized by HPLC, matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS), sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and trinitrobenzene sulfonic acid (TNBS). The secondary structure of the mPEG-ylated insulins was studied by circular Dichroism (CD). It is found that the secondary structure of insulin was changed hardly after mPEGylation. The in vivo experiment results showed that mPEG-ButyrALD-INS displayed longer half-life and extended action profiles.

Key words: insulin, mPEG, modification, secondary structure, hypoglycemic effect