Department of Chemistry and Biochemistry and California NanoSystems Institute, University of California, Los Angeles, 607 Charles E. Young Drive East, Los Angeles, California 90095-1569, United States.
J Am Chem Soc. 2022 Apr 6;144(13):6050-6058. doi: 10.1021/jacs.2c01136. Epub 2022 Mar 24.
Traceless self-immolative linkers are widely used for the reversible modification of proteins and peptides. This article describes a new class of traceless linkers based on or -hydroxybenzylamines. The introduction of electron-donating substituents on the aromatic core stabilizes the quinone methide intermediate, thus providing a platform for payload release that can be modulated. To determine the extent to which the electronics affect the rate of release, we prepared a small library of hydroxybenzylamine linkers with varied electronics in the aromatic core, resulting in half-lives ranging from 20 to 144 h. Optimization of the linker design was carried out with mechanistic insights from density functional theory (DFT) and the design of an intramolecular trapping agent through the use of DFT and intramolecular distortion energy calculations. This resulted in the development of a faster self-immolative linker with a half-life of 4.6 h. To demonstrate their effectiveness as traceless linkers for bioconjugation, reversible protein-polyethylene glycol conjugates with a model protein lysozyme were prepared, which had reduced protein activity but recovered ≥94% activity upon traceless release of the polymer. This new class of linkers with tunable release rates expands the traceless linkers toolbox for a variety of bioconjugation applications.
无痕迹自毁连接子广泛用于蛋白质和肽的可逆修饰。本文描述了一类基于邻-或对-羟苄胺的新的无痕迹连接子。在芳核上引入供电子取代基稳定了醌亚甲醚中间体，从而为可调节的有效载荷释放提供了一个平台。为了确定电子效应对释放速率的影响程度，我们制备了一系列具有不同电子性质的羟苄胺连接子的小分子库，其半衰期范围为 20 至 144 小时。通过密度泛函理论（DFT）的机制见解和使用 DFT 及分子内扭曲能计算设计的分子内捕获剂，对连接子设计进行了优化。这导致开发出一种半衰期为 4.6 小时的更快的自毁连接子。为了证明它们作为生物偶联无痕迹连接子的有效性，我们用模型蛋白溶菌酶制备了可逆的蛋白质-聚乙二醇缀合物，其蛋白质活性降低，但在聚合物无痕迹释放后恢复了≥94%的活性。这种具有可调释放速率的新型连接子扩展了无痕迹连接子工具箱，可用于各种生物偶联应用。
