Hirotsu Tatsunori, Higashi Taishi, Abu Hashim Irhan Ibrahim, Misumi Shogo, Wada Koki, Motoyama Keiichi, Arima Hidetoshi
Graduate School of Pharmaceutical Sciences, Kumamoto University , 5-1 Oe-honmachi, Chuo-ku, Kumamoto 862-0973, Japan.
Program for Leading Graduate Schools "HIGO (Health life science: Interdisciplinary and Glocal Oriented) Program", Kumamoto University , 5-1 Oe-honmachi, Chuo-ku, Kumamoto 862-0973, Japan.
Mol Pharm. 2017 Feb 6;14(2):368-376. doi: 10.1021/acs.molpharmaceut.6b00678. Epub 2017 Jan 12.
Polyethylene glycol (PEG) modification (PEGylation) is one of the best approaches to improve the stabilities and blood half-lives of protein drugs; however, PEGylation dramatically reduces the bioactivities of protein drugs. Here, we present "self-assembly PEGylation retaining activity" (SPRA) technology via a host-guest interaction between PEGylated β-cyclodextrin (PEG-β-CyD) and adamantane-appended (Ad) proteins. PEG-β-CyD formed stable complexes with Ad-insulin and Ad-lysozyme to yield SPRA-insulin and SPRA-lysozyme, respectively. Both SPRA-proteins showed high stability against heat and trypsin digest, comparable with that of covalently PEGylated protein equivalents. Importantly, the SPRA-lysozyme possessed ca. 100% lytic activity, whereas the activity of the covalently PEGylated lysozyme was ca. 23%. Additionally, SPRA-insulin provided a prolonged and peakless blood glucose profile when compared with insulin glargine. It also showed no loss of activity. In contrast, the covalently PEGylated insulin showed a negligible hypoglycemic effect. These findings indicate that SPRA technology has potential as a generic method, surpassing conventional PEGylation methods for proteins.
聚乙二醇（PEG）修饰（聚乙二醇化）是提高蛋白质药物稳定性和血液半衰期的最佳方法之一；然而，聚乙二醇化会显著降低蛋白质药物的生物活性。在此，我们通过聚乙二醇化β-环糊精（PEG-β-CyD）与金刚烷修饰的（Ad）蛋白质之间的主客体相互作用，提出了“保留活性的自组装聚乙二醇化”（SPRA）技术。PEG-β-CyD与Ad-胰岛素和Ad-溶菌酶形成稳定的复合物，分别产生SPRA-胰岛素和SPRA-溶菌酶。两种SPRA-蛋白质对热和胰蛋白酶消化均表现出高稳定性，与共价聚乙二醇化的等效蛋白质相当。重要的是，SPRA-溶菌酶具有约100%的裂解活性，而共价聚乙二醇化溶菌酶的活性约为23%。此外，与甘精胰岛素相比，SPRA-胰岛素提供了延长且无峰值的血糖曲线。它也没有活性损失。相比之下，共价聚乙二醇化胰岛素的降血糖作用可忽略不计。这些发现表明，SPRA技术作为一种通用方法具有潜力，优于传统的蛋白质聚乙二醇化方法。
