Analysis of functionalization of methoxy-PEG as maleimide-PEG
Analysis of functionalization of methoxy-PEG as maleimide-PEG
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DOI：
10.1016/j.ab.2007.11.034
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发表时间：
2008-03-15
影响因子：
2.9
通讯作者：
Manjula, Belur N.
中科院分区：
文献类型：
--
作者：
Ananda, K.;Nacharaju, Parimala;Manjula, Belur N.
蛋白质的伸展臂促进聚乙二醇化(EAFP)的设计利用了硫醇-马来酰亚胺反应的高选择性和定量方面的优势。然而，不同批次的马来酰亚胺-聚乙二醇酯对血红蛋白的EAFP效率不同。作为马来酰亚胺-聚乙二醇单甲氧基聚乙二醇酯(Mpeg)的低水平官能化已被研究为这种变异的潜在来源。利用谷胱甘肽、二硫苏糖醇和血红蛋白的硫醇基团与马来酰亚胺-聚乙二醇酯的反应，发展了一种新的化学方法来估计mpeg的官能化。对马来酰亚胺苯基异氰酸酯与聚乙二醇醚缩合的一步模块化合成方法进行了优化，得到了总纯度为80%的产品。核磁共振方法与新的化学方法所作的估计有很好的相关性。通过多步合成的商用马来酰亚胺-聚乙二醇酯试剂的官能化程度较低，这些化学估计反映了这一点。MPPEG功能化程度越高，EAFP的效率就越高，这主要体现在六聚乙二醇化Hb的生成和红细胞D抗原的掩蔽上。这种新的EAFP方案有望提高六聚乙二醇化血红蛋白、聚乙二醇化白蛋白和聚乙二醇化红细胞作为新的聚乙二醇化疗法的产生的成本效益。由爱思唯尔公司出版。
The design of the extension arm-facilitated PEGylation (EAFP) of proteins takes advantage of the high selective and quantitative aspects of the thiol-maleimide reaction. However, the efficiency of EAFP with hemoglobin varied with the batches of maleimide-PEG. The low level of functionalization of monomethoxy-PEG (mPEG) as maleimide-PEG has been now investigated as the potential source of this variation. New chemical approaches for the estimation of the functionalization of mPEG using the reaction of the thiol groups of glutathione, dithiothreitol, and hemoglobin with maleimide-PEG have been developed. The single-step modular approach to the synthesis of maleimidophenyl-PEG (MPPEG) that involved the condensation of p-maleimidophenyl isocyanate with mPEG has been optimized to generate a product with an overall purity of 80%. The NMR approach correlates well with the estimates made by the new chemical approaches. Commercial maleimide-PEG reagents synthesized using multiple steps exhibited a lower level of functionalization as reflected by these chemical estimations. The better functionalization of MPPEG increases the efficiency of EAFP as reflected by the generation of hexaPEGylated Hb and the masking of the D antigen of RBCs. This new EAFP protocol is expected to improve the cost effectiveness of the generation of hexaPEGylated Hb, PEGylated albumin, and PEGylated RBCs as new PEGylated therapeutics. Published by Elsevier Inc.