Maki Yuta, Mong Surin K, Chandrashekar Chaitra, Forbes Briony E, Hossain Mohammed Akhter, Yamaguchi Shintaro, Fadzen Colin M, Kajihara Yasuhiro, Pentelute Bradley L
Department of Chemistry, Graduate School of Science, Osaka University 1-1 Machikaneyama Toyonaka Osaka 560-0043 Japan
Forefront Research Center, Graduate School of Science, Osaka University 1-1 Machikaneyama Toyonaka Osaka 560-0043 Japan.
Chem Sci. 2025 Apr 4;16(18):7929-7935. doi: 10.1039/d5sc01670c. eCollection 2025 May 7.
Insulin is a key life-saving drug for patients with diabetes and is used clinically worldwide. To address the physicochemical challenges of insulin, such as low solubility and aggregation, glycosylated insulins have been chemically synthesized, exhibiting improved stability due to the hydration effect of glycans. In this work, we demonstrated the rapid synthesis of glycosylated insulins (glycoinsulins) using flow-based solid-phase peptide synthesis (SPPS). The insulin A-chain and glycosylated B-chain were synthesized by flow-based SPPS, with each elongation cycle completed in just 3 minutes. Through our investigations, the glycosylation step was successfully performed within 10 minutes under optimized flow-based conditions. Additionally, we examined the incorporation of dipeptide units (isoacyl dipeptide and pseudoproline) under flow conditions and demonstrated efficient peptide elongation by combining flow-based SPPS with these dipeptide units. The synthesized A- and B-chains were subsequently used for the stepwise formation of disulfide bond linkages. The resulting glycoinsulins exhibited comparable binding affinities to insulin receptors. These findings highlight a novel flow-based approach for the rapid synthesis of glycosylated peptide and protein drugs.
胰岛素是糖尿病患者的一种关键救命药物，在全球临床中广泛使用。为应对胰岛素的物理化学挑战，如低溶解度和聚集问题，已化学合成了糖基化胰岛素，由于聚糖的水合作用，其稳定性有所提高。在这项工作中，我们展示了使用基于流动的固相肽合成（SPPS）快速合成糖基化胰岛素（糖胰岛素）。胰岛素A链和糖基化B链通过基于流动的SPPS合成，每个延伸循环仅需3分钟即可完成。通过我们的研究，在优化的基于流动的条件下，糖基化步骤在10分钟内成功完成。此外，我们研究了在流动条件下二肽单元（异酰基二肽和假脯氨酸）的掺入情况，并通过将基于流动的SPPS与这些二肽单元相结合，证明了有效的肽延伸。随后，合成的A链和B链用于逐步形成二硫键连接。所得糖胰岛素对胰岛素受体表现出相当的结合亲和力。这些发现突出了一种基于流动的新方法，用于快速合成糖基化肽和蛋白质药物。
