Department of Chemistry, University of Copenhgen, Thorvaldsensvej 40, DK-1871, Frederiksberg, Denmark.
Current address: Faculty for Chemistry and Pharmacy, Ludwig-Maximilian University, Butenandtstr 5-13, 81377, Munich, Germany.
Chembiochem. 2024 Nov 18;25(22):e202400534. doi: 10.1002/cbic.202400534. Epub 2024 Oct 24.
Chemical modification of proteins is of growing importance to generate new molecular probes for chemical biology and for the development of new biopharmaceuticals. For example, two approved, long-acting insulin variants are lipidated at the LysB29 side-chain. Acylations of proteins have so far been performed in batch-mode. Here we describe the use of flow chemistry for site-selective acylation of a small protein, insulin. To the best of our knowledge this is the first report on flow chemistry for chemical modification of insulin. The first step was to develop reaction conditions for acylation of Lys B29 that gave a soluble mixture and thus was compatible with flow chemistry in a microreactor; this included selection of a soluble base. Secondly, the conditions, such as reagent ratios and flow rate were optimized. Third, the use of these conditions for the acylation with a wide range of acids was demonstrated. Finally, Boc-protected insulins were synthesized. Insulin remained stable towards these flow chemistry conditions. This use of flow chemistry for the chemical modification of insulin opens the prospect of producing chemically modified biopharmaceuticals by flow chemistry with fewer byproducts.
蛋白质的化学修饰对于生成新的化学生物学分子探针和开发新的生物制药具有重要意义。例如，两种已批准的长效胰岛素变体在赖氨酸 B29 侧链上发生脂质化。迄今为止，蛋白质的酰化反应一直是在批量模式下进行的。在这里，我们描述了使用流动化学对小蛋白胰岛素进行位点选择性酰化的方法。据我们所知，这是关于胰岛素化学修饰的流动化学的第一份报告。第一步是开发用于酰化赖氨酸 B29 的反应条件，该条件得到了可溶的混合物，因此与微反应器中的流动化学相容；这包括选择可溶性碱。其次，优化了试剂比和流速等条件。第三，展示了这些条件在与各种酸的酰化中的应用。最后，合成了 Boc 保护的胰岛素。胰岛素对这些流动化学条件保持稳定。这种使用流动化学对胰岛素进行化学修饰的方法为通过流动化学生产化学修饰的生物制药开辟了前景，可减少副产物的生成。
