A Traceless Site‐Specific Conjugation on Native Antibodies Enables Efficient One‐Step Payload Assembly
A Traceless Site‐Specific Conjugation on Native Antibodies Enables Efficient One‐Step Payload Assembly
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无痕位点 — 天然抗体的特异性缀合可实现高效的 One — Step 有效负载组装
DOI：
10.1002/ange.202204132
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发表时间：
2022
期刊：
影响因子：
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通讯作者：
Wei Huang
中科院分区：
文献类型：
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作者：
Yue Zeng;Wei Shi;Qian Dong;Wanzhen Li;Jianxin Zhang;Xuelian Ren;Caihong Tang;Bo Liu;Yuanli Song;Yali Wu;Xingxing Diao;Hu Zhou;He Huang;Feng Tang;Wei Huang
以位点特异性方式对天然抗体进行直接化学修饰仍然是一个巨大的挑战。配体定向缀合可以实现抗体的选择性修饰，但通常需要多个额外的步骤用于配体释放和货物组装。在此，我们报告了一种新的、无痕的策略，通过利用基于硫酯的酰基转移试剂来实现位点特异性抗体-药物缀合物（ADC）的简便且有效的一步合成。设计的试剂由优化的Fc靶向配体、硫酯桥和毒素有效载荷组成，直接将毒素精确组装到天然IgG的K251位置上，同时在一个步骤中自释放亲和配体。用这种方法，我们从天然曲妥珠单抗合成了一系列K251连接的ADC。这些ADC表现出优异的均一性、热稳定性以及体外和体内抗肿瘤活性。该策略对于IgG 1、IgG 2和IgG 4亚型同样有效。
Direct chemical modification of native antibodies in a site-specific manner remains a great challenge. Ligand-directed conjugation can achieve the selective modification of antibodies, but usually requires multiple extra steps for ligand release and cargo assembly. Herein, we report a novel, traceless strategy to enable the facile and efficient one-step synthesis of site-specific antibody-drug conjugates (ADCs) by harnessing a thioester-based acyl transfer reagent. The designed reagent, consisting of an optimized Fc-targeting ligand, a thioester bridge and a toxin payload, directly assembles the toxin precisely onto the K251 position of native IgGs and simultaneously self-releases the affinity ligand in one step. With this method, we synthesized a series of K251-linked ADCs from native Trastuzumab. These ADCs demonstrated excellent homogeneity, thermal stability, and both in vitro and in vivo anti-tumor activity. This strategy is equally efficient for IgG1, IgG2, and IgG4 subtypes.