马来酰亚胺
化学
生物结合
结合
组合化学
共价键
Diels-Alder反应
加合物
体内
有机化学
催化作用
数学
生物
数学分析
生物技术
作者
André H. St. Amant,Feng‐Ying Huang,Jianqiang Lin,Daniel Lemen,Chacko Chakiath,Shenlan Mao,Christine Fazenbaker,Haihong Zhong,Jay Harper,Wenshu Xu,Neki Patel,Lauren Adams,Balakumar Vijayakrishnan,Philip W. Howard,Marcello Marelli,Herren Wu,Changshou Gao,Javier Read de Alaniz,R. James Christie
标识
DOI:10.1021/acs.bioconjchem.9b00436
摘要
The normal electron-demand Diels–Alder (DA) cycloaddition is a classic transformation routinely used in synthesis; however, applications in biological systems are limited. Here, we report a spiro[2.4]hepta-4,6-diene-containing noncanonical amino acid (SCpHK) capable of efficient incorporation into antibodies and subsequent coupling with maleimide via a DA reaction. SCpHK was stable throughout protein expression in mammalian cells and enabled covalent attachment of maleimide drug-linkers yielding DA antibody–drug conjugates (DA-ADCs) with nearly quantitative conversion in a one-step process. The uncatalyzed DA reaction between SCpHK and maleimide in aqueous buffer was rapid (1.8–5.4 M–1 s–1), and the antibody–drug adduct was stable in rat serum for at least 1 week at 37 °C. Anti-EphA2 DA-ADCs containing AZ1508 or SG3249 maleimide drug-linkers were potent inhibitors of tumor growth in PC3 tumor models in vivo. The DA bioconjugation strategy described here represents a simple method to produce site-specific and stable ADCs with maleimide drug-linkers.
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