Supramolecular Protein Stabilization with Zwitterionic Polypeptide–Cucurbit[7]uril Conjugates
Supramolecular Protein Stabilization with Zwitterionic Polypeptide–Cucurbit[7]uril Conjugates
复制标题
使用两性离子多肽 – 葫芦[7]脲缀合物实现超分子蛋白质稳定性
DOI：
10.1021/acs.biomac.2c01319
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发表时间：
2023
影响因子：
6.2
通讯作者：
Kramer, Jessica R.
中科院分区：
文献类型：
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作者：
Clauss, Zachary S.;Meudom, Rolande;Su, Bo;VandenBerg, Michael A.;Saini, Simranpreet S.;Webber, Matthew J.;Chou, Danny Hung-Chieh;Kramer, Jessica R.
蛋白质聚集是开发新生物药物的障碍，对重要疗法的运输和储存提出了挑战。尽管已经探索了各种材料和方法，但仍然需要一种简单的材料，该材料是可生物降解的、无毒的并且在稳定蛋白质治疗剂方面高效。在这项工作中，我们研究了使用快速和可扩展的聚合技术制备的两性离子多肽，并与超分子大环主体葫芦[7]脲缀合，以抑制模型蛋白治疗剂胰岛素和降钙素聚集的能力。多肽基于天然氨基酸甲硫氨酸，并将两性锍修饰与类似的阳离子和中性结构进行比较。每个聚合物都用单个葫芦[7]脲大环进行末端修饰，以提供超分子识别并与蛋白质上的末端芳香族氨基酸结合。只有从足够链长的两性离子结构制备的缀合物才是胰岛素聚集的有效抑制剂，并且也可以抑制降钙素的聚集。该多肽在人细胞中甚至在预期治疗方案的五倍的浓度下也没有表现出细胞毒性。我们探索了用一组天然蛋白酶处理两性离子多肽，并发现如预期的稳定生物降解，支持当用作蛋白质制剂添加剂时的最终清除。
Protein aggregation is an obstacle for the development of new biopharmaceuticals, presenting challenges in shipping and storage of vital therapies. Though a variety of materials and methods have been explored, the need remains for a simple material that is biodegradable, nontoxic, and highly efficient at stabilizing protein therapeutics. In this work, we investigated zwitterionic polypeptides prepared using a rapid and scalable polymerization technique and conjugated to a supramolecular macrocycle host, cucurbit[7]uril, for the ability to inhibit aggregation of model protein therapeutics insulin and calcitonin. The polypeptides are based on the natural amino acid methionine, and zwitterion sulfonium modifications were compared to analogous cationic and neutral structures. Each polymer was end-modified with a single cucurbit[7]uril macrocycle to afford supramolecular recognition and binding to terminal aromatic amino acids on proteins. Only conjugates prepared from zwitterionic structures of sufficient chain lengths were efficient inhibitors of insulin aggregation and could also inhibit aggregation of calcitonin. This polypeptide exhibited no cytotoxicity in human cells even at concentrations that were five-fold of the intended therapeutic regime. We explored treatment of the zwitterionic polypeptides with a panel of natural proteases and found steady biodegradation as expected, supporting eventual clearance when used as a protein formulation additive.