In situ growth of a PEG-like polymer from the C terminus of an intein fusion protein improves pharmacokinetics and tumor accumulation
In situ growth of a PEG-like polymer from the C terminus of an intein fusion protein improves pharmacokinetics and tumor accumulation
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DOI：
10.1073/pnas.1006044107
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发表时间：
2010-09-21
影响因子：
11.1
通讯作者：
Chilkoti, Ashutosh
中科院分区：
文献类型：
--
作者：
Gao, Weiping;Liu, Wenge;Chilkoti, Ashutosh
本文报道了一种仅从重组蛋白的C末端生长聚合物缀合物的通用原位方法。GFP在其C末端与内含肽融合;内含肽的切割在GFP的C末端提供了独特的硫酯部分，其用于安装原子转移自由基聚合（ATRP）引发剂。随后在原位ATRP的寡（乙二醇）甲基醚甲基丙烯酸酯（OEGMA）产生了一个位点特异性（C-末端）和化学计量共轭高产率和良好的保留蛋白质的活性。GFP-C-聚（OEGMA）缀合物（流体动力学半径（R（h））：21 nm）显示，在静脉内给予小鼠后，与蛋白质（R（h）：3.0 nm）相比，其血液暴露增加了15倍。与未修饰的蛋白质相比，该缀合物还显示在静脉内施用给荷瘤小鼠后24小时肿瘤积累增加50倍。这种用于重组蛋白的原位C-末端聚合物修饰的方法适用于重组蛋白和肽药物的大子集，并提供了用于改善其药理学特征的一般方法。
This paper reports a general in situ method to grow a polymer conjugate solely from the C terminus of a recombinant protein. GFP was fused at its C terminus with an intein; cleavage of the intein provided a unique thioester moiety at the C terminus of GFP that was used to install an atom transfer radical polymerization (ATRP) initiator. Subsequent in situ ATRP of oligo(ethylene glycol) methyl ether methacrylate (OEGMA) yielded a site-specific (C-terminal) and stoichiometric conjugate with high yield and good retention of protein activity. A GFP-C-poly(OEGMA) conjugate (hydrodynamic radius (R(h)): 21 nm) showed a 15-fold increase in its blood exposure compared to the protein (R(h): 3.0 nm) after intravenous administration to mice. This conjugate also showed a 50-fold increase in tumor accumulation, 24 h after intravenous administration to tumor-bearing mice, compared to the unmodified protein. This approach for in situ C-terminal polymer modification of a recombinant protein is applicable to a large subset of recombinant protein and peptide drugs and provides a general methodology for improvement of their pharmacological profiles.