Structure-Based Site-Specific PEGylation of Fibroblast Growth Factor 2 Facilitates Rational Selection of Conjugate Sites
Structure-Based Site-Specific PEGylation of Fibroblast Growth Factor 2 Facilitates Rational Selection of Conjugate Sites
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成纤维细胞生长因子 2 基于结构的位点特异性聚乙二醇化有利于缀合位点的合理选择
DOI：
10.1002/biot.201900203
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发表时间：
2020
影响因子：
4.7
通讯作者：
Jianlou Niu
中科院分区：
文献类型：
--
作者：
Jing Zhao;Qi Li;Jiamin Wu;Chuanren Zhou;Yu Cao;Xiaokun Li;Jianlou Niu
聚乙二醇修饰（PEGylation）可以通过将聚乙二醇（PEG）附着在蛋白质表面来保护蛋白质表面免受蛋白质水解降解和限制聚集，从而增强治疗蛋白的药代动力学特性。然而，目前的聚乙二醇化策略往往会降低生物活性，这可能是由于聚乙二醇的空间位阻。总的来说，目前还没有基于结构的结合位点选择指南，以保持最佳的生物活性和改善的药代动力学特性。在这项研究中，基于FGF2 - FGFR1 -肝素复合物结构进行了位点特异性聚乙二醇化。通过测量修饰蛋白的受体/肝素结合亲和力，并进行基于细胞的生物活性和体内稳定性的测定，研究了偶联位点对蛋白质功能的影响。这些数据的综合分析表明，FGF2的聚乙二醇化避免了成纤维细胞生长因子受体1 （FGFR1）和肝素的结合位点，提供了最佳的药代动力学增强，生物活性损失最小。动物实验表明，与未修饰的FGF2相比，聚乙二醇化的FGF2在防止创伤性脑损伤诱导的脑损伤和神经功能方面表现出更大的功效。这种基于合理结构的聚乙二醇化蛋白质修饰策略预计将在基于蛋白质的治疗领域产生重大影响。
Polyethylene glycol modification (PEGylation) can enhance the pharmacokinetic properties of therapeutic proteins by the attachment of polyethylene glycol (PEG) to the surface of a protein to shield the protein surface from proteolytic degradation and limit aggregation. However, current PEGylation strategies often reduce biological activity, potentially as a result of steric hindrance of PEG. Overall, there are no structure‐based guidelines for selection of conjugate sites that retain optimal biological activity with improved pharmacokinetic properties. In this study, site‐specific PEGylation based on the FGF2‐FGFR1‐heparin complex structure is performed. The effects of the conjugate sites on protein function are investigated by measuring the receptor/heparin binding affinities of the modified proteins and performing assays to measure cell‐based bio‐activity and in vivo stability. Comprehensive analysis of these data demonstrates that PEGylation of FGF2 that avoids the binding sites for fibroblast growth factor receptor 1 (FGFR1) and heparin provides optimal pharmacokinetic enhancement with minimal losses to biological activity. Animal experiments demonstrate that PEGylated FGF2 exhibits greater efficacy in protecting against traumatic brain injury‐induced brain damage and neurological functions than the non‐modified FGF2. This rational structure‐based PEGylation strategy for protein modification is expected to have a major impact in the area of protein‐based therapeutics.