Zhejiang Provincial Key Laboratory of Anti-Cancer Drug Research, Institute of Drug Metabolism and Drug Analysis, College of Pharmaceutical Sciences, Zhejiang University, Hangzhou 310058, China.
College of Pharmaceutical Sciences, Zhejiang Chinese Medical University, Hangzhou 310053, China.
Int J Mol Sci. 2017 Oct 31;18(11):2284. doi: 10.3390/ijms18112284.
The therapeutic efficacy of antibodies can be successfully improved through targeted delivery of potent cytotoxic drugs in the form of antibody-drug conjugates. However, conventional conjugation strategies lead to heterogeneous conjugates with undefined stoichiometry and sites, even with considerable batch-to-batch variability. In this study, we have developed a chemo-enzymatic strategy by equipping the C-terminus of anti-CD20 ofatumumab with a click handle using Sortase A, followed by ligation of the payload based on a strain-promoted azide-alkyne cycloaddition to produce homogeneous conjugates. The resulting antibody-drug conjugates fully retained their antigen binding capability and proved to be internalized and trafficked to the lysosome, which released the payload with a favorable efficacy in vitro and in vivo. Thus, this reported method is a versatile tool with maximum flexibility for development of antibody-drug conjugates and protein modification.
通过以抗体药物偶联物的形式靶向递送有效细胞毒性药物，抗体的治疗效果可以得到成功改善。然而，传统的偶联策略会导致不均一的偶联物，具有不确定的化学计量和连接点，即使在批间变化很大的情况下也是如此。在这项研究中，我们通过使用 Sortase A 在抗 CD20 的奥法木单抗的 C 末端装备一个点击手柄，开发了一种化学酶策略，然后根据应变促进的叠氮-炔环加成反应将有效载荷连接起来，以产生均一的偶联物。所得的抗体药物偶联物完全保留了其抗原结合能力，并被证明能够内化并转运到溶酶体，在体外和体内都显示出良好的疗效。因此，该报道的方法是一种具有最大灵活性的通用工具，可用于开发抗体药物偶联物和蛋白质修饰。
