Gao Han, Wang Si-Tao, Wang Haibin, Fang Wei-Jie
Institute of Drug Metabolism and Pharmaceutical Analysis, College of Pharmaceutical Sciences, Zhejiang University, Hangzhou 310058, China; Hangzhou Institute of Innovative Medicine, Zhejiang University, Hangzhou 310016, China.
Institute of Drug Metabolism and Pharmaceutical Analysis, College of Pharmaceutical Sciences, Zhejiang University, Hangzhou 310058, China.
J Pharm Biomed Anal. 2023 May 10;228:115347. doi: 10.1016/j.jpba.2023.115347. Epub 2023 Mar 15.
For monoclonal antibody (mAb) drugs, the 'me-too' drug is a pharmacologically active compound that is structurally similar to the first-in-class drugs, acting on the same target and is used for the same therapeutic purposes, but it may differ in drug-drug interactions and adverse drug reactions. Capillary electrophoresis-sodium dodecyl sulfate (CE-SDS) has been widely used for quality evaluation of mAb drugs. The properties of the detected substances can interfere with the credibility and accuracy of the method. In the routine comparison analysis for both innovator rituximab and 'me-too' drug zuberitamab samples, an uncommon artifact related to the heavy chain (HC) of zuberitamab was observed in reducing CE-SDS and interfered with our identification of the purity of samples. In this work, the overall hydrophobicity of the HCs of rituximab, zuberitamab, and several other common mAbs was characterized and determined by reversed-phase high-performance liquid chromatography. Additionally, the local hydrophobicity and surface charge were compared using Expasy ProtScale and PyMOL software simulations. We concluded that noncovalent protein aggregation can be related to strong hydrophobicity and low electrostatic repulsion of local amino acid regions, which complicates drug quality control. These findings shed light on the relationship between protein aggregation and the local hydrophobicity region, and broaden the way to analyze the detection 'artifacts' in reducing CE-SDS studies of therapeutic proteins.
对于单克隆抗体（mAb）药物，“me-too”药物是一种药理活性化合物，其结构与同类首创药物相似，作用于相同靶点且用于相同治疗目的，但在药物相互作用和药物不良反应方面可能存在差异。毛细管电泳 - 十二烷基硫酸钠（CE - SDS）已广泛用于mAb药物的质量评估。被检测物质的性质会干扰该方法的可信度和准确性。在对创新药利妥昔单抗和“me-too”药物泽布替尼单抗样品的常规比较分析中，在还原性CE - SDS中观察到一种与泽布替尼单抗重链（HC）相关的罕见假象，干扰了我们对样品纯度的鉴定。在这项工作中，通过反相高效液相色谱对利妥昔单抗、泽布替尼单抗和其他几种常见mAb的重链的整体疏水性进行了表征和测定。此外，使用Expasy ProtScale和PyMOL软件模拟比较了局部疏水性和表面电荷。我们得出结论，非共价蛋白质聚集可能与局部氨基酸区域的强疏水性和低静电排斥有关，这使药物质量控制变得复杂。这些发现揭示了蛋白质聚集与局部疏水性区域之间的关系，并拓宽了在治疗性蛋白质的还原性CE - SDS研究中分析检测“假象”的途径。
