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Development of polymeric albumin preparation with prolonged half-life by redox reaction

利用氧化还原反应开发半衰期延长的聚合白蛋白制剂

基本信息

批准号：
19790130
负责人：
IWAO Yasunori
金额：
$ 2.41万
依托单位：
University of Shizuoka (2008-2009)Kumamoto University (2007)
依托单位国家：
日本
项目类别：
Grant-in-Aid for Young Scientists (B)
财政年份：
2007
资助国家：
日本
起止时间：
2007 至 2009
项目状态：
已结题

项目摘要

In this study, we aim to develop a novel polymeric albumin preparation with prolonged half-life by using site-directed mutagenesis and redox reaction. First, to obtain fundamental information regarding which amino acid residues in albumin are involved in its elimination, the pharmacokinetics and antioxidant activity of 26 genetic variants of albumin, as well as oxidized and glycated albumins were examined. As a result, we found that when two amino acid residues, namely, Glu 570 and Lys 573, in domain III were mutated to Arg, one Arg was added to the N-terminal and domain II were glycosylated, the half-life of albumin was increased. On the basis of these results, we performed multiple mutations to albumin and the mutant was further polymerized. The polymeric recombinant albumin mutant was correctly folded, as evidenced by the similarity of the patterns in the far- and near-UV circular dichroism spectra of mutant and native albumin. In addition, the plasma levels of the polymeric albumin mutant after intravenous bolus injection to rats exhibited two-phase profiles, and the half-life was longer than that of native albumin. Furthermore, the polymeric albumin showed a reduction in edema against paw edema mouse models because of its high molecular weight. These results indicate the potential for development of the new polymeric recombinant albumin preparation with prolonged half-life, but further investigation is needed to ensure the safety of the albumin preparation.

在这项研究中，我们的目标是通过定点诱变和氧化还原反应开发一种具有延长半衰期的新型聚合白蛋白制剂。首先，为了获得白蛋白中哪些氨基酸残基参与白蛋白消除的基本信息，我们检查了 26 种白蛋白遗传变体以及氧化白蛋白和糖化白蛋白的药代动力学和抗氧化活性。结果，我们发现，当结构域III中的两个氨基酸残基，即Glu 570和Lys 573突变为Arg，N端添加一个Arg并且结构域II被糖基化时，白蛋白的半衰期增加。根据这些结果，我们对白蛋白进行了多次突变，并将突变体进一步聚合。聚合重组白蛋白突变体被正确折叠，突变体和天然白蛋白的远紫外和近紫外圆二色光谱模式的相似性证明了这一点。此外，对大鼠静脉推注后，聚合白蛋白突变体的血浆水平呈现两相分布，且半衰期比天然白蛋白更长。此外，由于其高分子量，聚合白蛋白在爪水肿小鼠模型中显示出水肿减轻。这些结果表明开发具有延长半衰期的新型聚合重组白蛋白制剂的潜力，但需要进一步研究以确保白蛋白制剂的安全性。