Zwitterlation mitigates protein bioactivity loss in vitro over PEGylation
Zwitterlation mitigates protein bioactivity loss in vitro over PEGylation
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DOI：
10.1039/c8sc01777h
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发表时间：
2018-12-07
期刊：
影响因子：
8.4
通讯作者：
Jiang, Shaoyi
中科院分区：
文献类型：
--
作者：
Han, Yanjiao;Yuan, Zhefan;Jiang, Shaoyi
与聚（乙二醇）（PEG）或PEG化的缀合是一种广泛使用的工具，以克服天然蛋白质的缺点，如稳定性差，药代动力学（PK）特征不足，和免疫原性。然而，PEG化通常伴随着对生物活性的不希望的有害影响，特别是由PEG的两亲性质引起的。这对于具有大结合靶标的聚乙二醇化蛋白质尤其如此。Pegasys，一种带有40 kDa支链PEG的PEG化干扰素α-2a（IFN-α 2a），是一个典型的例子，其仅显示出未修饰的IFN-α 2a的7%的体外活性。在这项工作中，通过采用IFN-α 2a作为模型蛋白，我们证明了与两性离子聚合物（或两性化）缀合的蛋白质可以显著减轻聚合物缀合后体外抗增殖生物活性的损失。两性化IFN-α 2a的保留抗增殖活性比具有相同聚合物分子量的PEG化IFN-α 2a高4.4倍，或比具有相似流体动力学尺寸的PEG化IFN-α 2a高3倍。假设两性离子聚合物和IFN-α 2a/IFN-α 2a受体之间的非特异性相互作用可以由于两性离子聚合物的超亲水性质而减轻。这反过来又减少了IFN-α 2a和IFN-α 2a受体之间的“非特异性阻断”。此外，我们证明了两性化IFN-α 2a在大鼠中重复注射后显示出延长的循环时间和减轻的加速血液清除。
Conjugation with poly(ethylene glycol) (PEG) or PEGylation is a widely used tool to overcome the shortcomings of native proteins, such as poor stability, inadequate pharmacokinetic (PK) profiles, and immunogenicity. However, PEGylation is often accompanied by an unwanted detrimental effect on bioactivity, particularly, resulting from the amphiphilic nature of PEG. This is especially true for PEGylated proteins with large binding targets. Pegasys, a PEGylated interferon alpha-2a (IFN-alpha 2a) bearing a 40 kDa branched PEG, is a typical example that displays only 7% in vitro activity of the unmodified IFN-alpha 2a. In this work, by employing IFN-alpha 2a as a model protein, we demonstrated that a protein conjugated with zwitterionic polymers (or zwitterlation) could significantly mitigate the antiproliferative bioactivity loss in vitro after polymer conjugation. The retained antiproliferative activity of zwitterlated IFN-alpha 2a is 4.4-fold higher than that of the PEGylated IFN-alpha 2a with the same polymer molecular weight, or 3-fold higher than that of the PEGylated IFN-alpha 2a with a similar hydrodynamic size. It is hypothesized that nonspecific interactions between zwitterionic polymers and IFN-alpha 2a/IFN-alpha 2a receptors can be mitigated due to the super-hydrophilic nature of zwitterionic polymers. This, in turn, reduces the 'nonspecific blocking' between IFN-alpha 2a and IFN-alpha 2a receptors. In addition, we demonstrated that zwitterlated IFN-alpha 2a showed a prolonged circulation time and a mitigated accelerated blood clearance after repeated injections in rats.