One-step synthesis of site-specific antibody-drug conjugates by reprograming IgG glycoengineering with LacNAc-based substrates  

作　　者：Wei Shi Wanzhen Li Jianxin Zhang Tiehai Li Yakai Song Yue Zeng Qian Dong Zeng Lin Likun Gong Shuquan Fan Feng Tang Wei Huang 

机构地区：[1]CAS Key Laboratory of Receptor Research,CAS Center for Excellence in Molecular Cell Science,Center for Biotherapeutics Discovery Research,Shanghai Institute of Materia Medica Chinese Academy of Sciences,Shanghai 201203,China [2]School of Pharmaceutical Science and Technology,Hangzhou Institute of Advanced Study,Hangzhou 310024,China [3]University of Chinese Academy of Sciences,Beijing 100049,China [4]School of Chinese Materia Medica,Nanjing University of Chinese Medicine,Nanjing 210023,China [5]School of Life Science,Liaocheng University,Liaocheng 252059,China

出　　处：《Acta Pharmaceutica Sinica B》2022年第5期2417-2428,共12页药学学报（英文版）

基　　金：supported by the National Natural Science Foundation of China(NSFC,No.,2187116 and 82003574);Shanghai Municipal Science and Technology Major Project,the Shanghai Sail Program(No.19YF1457100);the Special Research Assistant Program(Chinese Academy of Sciences,CAS),Natural Science Foundation of Shandong Province(ZR2017BC062)。

摘　　要：Glycosite-specific antibody-drug conjugatess(gsADCs), harnessing Asn297 N-glycan of IgG Fc as the conjugation site for drug payloads, usually require multi-step glycoengineering with two or more enzymes, which limits the substrate diversification and complicates the preparation process.Herein, we report a series of novel disaccharide-based substrates, which reprogram the IgG glycoengineering to one-step synthesis of gsADCs, catalyzed by an endo-N-acetylglucosaminidase(ENGase) of Endo-S2. IgG glycoengineering via ENGases usually has two steps: deglycosylation by wild-type(WT) ENGases and transglycosylation by mutated ENGases. But in the current method, we have found that disaccharide LacNAc oxazoline can be efficiently assembled onto IgG by WT Endo-S2 without hydrolysis of the product, which enables the one-step glycoengineering directly from native antibodies.Further studies on substrate specificity revealed that this approach has excellent tolerance on various modification of 6-Gal motif of LacNAc. Within 1 h, one-step synthesis of gsADC was achieved using the LacNAc-toxin substrates including structures free of bioorthogonal groups. These gsADCs demonstrated good homogeneity, buffer stability, in vitro and in vivo anti-tumor activity. This work presents a novel strategy using LacNAc-based substrates to reprogram the multi-step IgG glycoengineering to a one-step manner for highly efficient synthesis of gsADCs.

关 键 词：Site-specific ADCs ENGase LacNAc One-step assembly Potent in vivo efficacy 

分 类 号：R914[医药卫生—药物化学]