Global Research Technologies, Novo Nordisk Research Park, Måløv 2760, Denmark.
Carbon Dioxide Activation Center (CADIAC), Interdisciplinary Nanoscience Center, Department of Chemistry, Aarhus University, Aarhus C 8000, Denmark.
Bioconjug Chem. 2022 Jan 19;33(1):172-179. doi: 10.1021/acs.bioconjchem.1c00528. Epub 2021 Dec 28.
This paper describes the discovery, synthesis, and use of novel water-soluble acylation reagents for efficient and selective modification, cross-linking, and labeling of proteins and peptides, as well as for their use in the effective modification of sepharose beads under pH control in aqueous media. The reagents are based on a 2,4-dichloro-6-sulfonic acid phenol ester core combined with a variety of linker structures. The combination of these motifs leads to an ideal balance between hydrolytic stability and reactivity. At high pH, good to excellent conversions (up to 95%) and regioselectivity (up to 99:1 / amine ratio) in the acylation were realized, exemplified by the chemical modification of incretin peptides and insulin. At neutral pH, an unusually high preference toward the -terminal phenylalanine in an insulin derivative was observed (>99:1 /), which is up until now unprecedented in the literature for more elaborate reagents. In addition, the unusually high hydrolytic stability of these reagents and their ability to efficiently react at low concentrations (28 μM or 0.1 mg/mL) are exemplified with a hydroxy linker-based reagent and are a unique feature of this work.
本文描述了新型水溶性酰化试剂的发现、合成及应用，这些试剂可用于高效、选择性地修饰、交联和标记蛋白质和肽，以及在 pH 控制下，在水相介质中有效地修饰琼脂糖珠。这些试剂基于 2,4-二氯-6-磺酸苯酚酯核心，结合了多种连接子结构。这些结构的结合导致水解稳定性和反应性之间达到理想的平衡。在高 pH 值下，酰化反应具有良好到优异的转化率（高达 95%）和区域选择性（高达 99:1 / 胺比），以肠降血糖素肽和胰岛素的化学修饰为例。在中性 pH 值下，在胰岛素衍生物中观察到对 -末端苯丙氨酸的异常高选择性（>99:1 / ），这在更复杂的试剂中迄今为止是前所未有的。此外，以基于羟基连接子的试剂为例，这些试剂异常高的水解稳定性及其在低浓度（28 μM 或 0.1 mg/mL）下高效反应的能力是这项工作的独特特征。
