一种抗HER2人源化单克隆抗体药物的药学研究.docx

作者简介：王晓闻，女，硕士 研究方向：抗体药物质量分析评价 *通讯作者：谭青乔，男，博士 研究方向：抗体药物质量分析评价 Tel：（021E-mail: tanqq@ 一种抗HER2人源化单克隆抗体药物的药学研究 王晓闻，刘培培，吕锋华，谭青乔*（抗体药物国家工程研究中心，201203） 摘要：目的 对一种用于治疗HER2阳性乳腺癌的抗HER2人源化单克隆抗体靶向生物制剂进行有效性和安全性的药学研究。方法 采用高效液相色谱、毛细管电泳、圆二色谱、差示扫描量热技术、液质联用和基于人乳腺癌细胞的体外活性检测以及SPR技术分析抗HER2抗体的纯度、结构、翻译后修饰以及功能活性等关键质量属性。结果 该抗体的单体纯度98%，聚体含量2%；轻重链电泳纯度97%；具有正确的二级与三级结构，相变温度为72.65±0.1℃，和85.08±0.1℃；大于99%的分子具有正确的糖基化修饰，且N糖分布大体上与参比制剂高度相似，高甘露糖含量少但唾液酸含量多；对人乳腺癌细胞的增殖抑制活性和ADCC活性与参比制剂相当（P=0.96），与HER2抗原以及Fc受体的结合也无显著差异。结论 该抗体在药学评价上展现了高纯度、低杂质、结构稳定以及正确的翻译后修饰的特性，通过如此精心的分子和工艺设计，使其表现出与参比制剂相同的生物等效功能作用，而且也在免疫原性和稳定性上都表现出低风险，具备良好的安全性和有效性。 关键词: HER2; 单克隆抗体; 质量; 有效性; 安全性 The Pharmaceutical Research of an Anti-HER2 Humanized Monoclonal Antibody Drug Product WANG Xiao-wen, LIU Pei-pei, LV Feng-hua，TAN Qing-qiao*（National Engineering Research Center of Antibody Medicine，Shanghai 201203，China） ABSTRACT： OBJECTIVE To perform a pharmaceutical study on the efficacy and safety of a therapeutic anti-HER2 humanized monoclonal antibody targeting HER2-positive breast cancer. METHODS The critical quality attributes such as purity, structure, post-translation modifications, function and efficacy are analyzed using HPLC, CE-SDS, Circular Dichroism, Differential Scanning Calorimeter, LC-MS/MS, human breast cancer cell-based in vitro bioactivity assay and SPR binding kinetics. RESULTS The monomer content of the antibody is more than 98% while the polymer impurities are less than 2%. The sum of heavy chain and light chain peak area is over 97% on the CE-SDS electrophoretogram. The antibody drug demonstrates correct secondary and tertiary structure with two phase transition temperatures of 72.65±0.1℃ and 85.08±0.1℃. More than 99% of the molecules are correctly glycosylated with a highly similar N-glycan profiling to the reference product. The differences exist where the experiment subject has higher content of high mannose structure but lower sialic acids. The two products keep the same level on BT474 proliferation inhib