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

在这项研究中，我们的目标是通过定点突变和氧化还原反应来开发一种新型的半衰期延长的聚合物白蛋白制剂。首先，为了获得白蛋白中哪些氨基酸残基参与其消除的基本信息，研究了白蛋白的26个遗传变异以及氧化和糖化的白蛋白的药代动力学和抗氧化活性。结果发现，当结构域III的两个氨基酸残基，即Glu 570和Lys 573突变为Arg时，结构域II的N末端增加了一个Arg，糖基化，白蛋白的半衰期延长。根据这些结果，我们对白蛋白进行了多次突变，并对突变体进行了进一步聚合。突变体和天然白蛋白的远、近紫外圆二色谱图谱相似，证明重组白蛋白突变体能够正确折叠。此外，大鼠静脉注射聚合型白蛋白突变体后的血浆浓度呈双相分布，半衰期长于天然白蛋白。此外，聚合物白蛋白由于其高分子量，在小鼠足肿胀模型中显示出减轻肿胀的作用。这些结果表明新型聚合物重组白蛋白制剂具有开发潜力，半衰期延长，但需要进一步研究以确保白蛋白制剂的安全性。

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.