One-step synthesis of site-specific antibody-drug conjugates by reprograming IgG glycoengineering with LacNAc-based substrates.
One-step synthesis of site-specific antibody-drug conjugates by reprograming IgG glycoengineering with LacNAc-based substrates.
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通过使用基于 LacNAc 的底物重编程 IgG 糖工程一步合成位点特异性抗体-药物缀合物
DOI：
10.1016/j.apsb.2021.12.013
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发表时间：
2022-05
影响因子：
14.5
通讯作者：
Huang, Wei
中科院分区：
文献类型：
--
作者：
Shi, Wei;Li, Wanzhen;Zhang, Jianxin;Li, Tiehai;Song, Yakai;Zeng, Yue;Dong, Qian;Lin, Zeng;Gong, Likun;Fan, Shuquan;Tang, Feng;Huang, Wei
利用IgG Fc的Asn 297 N-聚糖作为药物有效载荷的缀合位点的糖位点特异性抗体药物缀合物（gsADC）通常需要用两种或更多种酶进行多步糖工程改造，这限制了底物多样化并使制备过程复杂化。在此，我们报告了一系列新的基于dislidamide的底物，其将IgG糖工程重新编程为gsADC的一步合成，由Endo-S2的内切-N-乙酰氨基葡萄糖苷酶（ENGase）催化。经由ENGase的IgG糖工程化通常具有两个步骤：通过野生型（WT）ENGase的去糖基化和通过突变ENGase的转糖基化。但是在目前的方法中，我们已经发现二糖LacNAc恶唑啉可以通过WT Endo-S2有效地组装到IgG上而不水解产物，这使得能够直接从天然抗体进行一步糖工程化。对底物特异性的进一步研究表明，该方法对LacNAc的6-Gal基序的各种修饰具有良好的耐受性。在1小时内，使用包括不含生物正交基团的结构的LacNAc-毒素底物实现gsADC的一步合成。这些gsADC表现出良好的均一性、缓冲液稳定性、体外和体内抗肿瘤活性。这项工作提出了一种新的策略，使用基于LacNAc的底物将多步IgG糖工程重新编程为一步方式，用于高效合成gsADC。基于LacNAc的底物对多步IgG糖工程进行重编程，并使得能够通过野生型内切糖苷酶（Endo-S2）催化有效地一步合成糖位点特异性抗体-药物缀合物（gsADC）。该方法还有利于生物素化、荧光标记、额外聚糖延伸等的多样性修饰以获得功能。
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. A LacNAc-based substrate reprograms the multi-step IgG glycoengineering and enables the efficient one-step synthesis of glycosite-specific antibody–drug conjugates (gsADCs), catalyzed by a wild-type endo-glycosidase (Endo-S2). This method also facilitates diverse modification of biotinylation, fluorescent labeling, extra glycan elongation, etc. for gain of functions.
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糖工程化 IgG 抗体和糖位点特异性抗体-药物缀合物的化学酶合成
影响因子：
14.8
作者：
Tang F;Wang LX;Huang W
通讯作者：
Huang W
影响因子：
4.9
作者：
Lhospice, F.;Bregeon, D.;Romagne, F.
通讯作者：
Romagne, F.
影响因子：
3.7
作者：
Beerli RR;Hell T;Merkel AS;Grawunder U
通讯作者：
Grawunder U
DOI：
10.1073/pnas.1702173114
发表时间：
2017-03-28
影响因子：
11.1
作者：
Li, Tiezheng;DiLillo, David J.;Wang, Lai-Xi
通讯作者：
Wang, Lai-Xi
影响因子：
4.7
作者：
van Geel, Remon;Wijdeven, Marloes A.;van Delft, Floris L.
通讯作者：
van Delft, Floris L.