Development and in vivo evaluation of an oral insulin-PEG delivery system
Development and in vivo evaluation of an oral insulin-PEG delivery system
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DOI：
10.1016/j.ejps.2004.03.015
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发表时间：
2004-07-01
影响因子：
4.6
通讯作者：
Bernkop-Schnürch, A
中科院分区：
文献类型：
--
作者：
Calceti, P;Salmaso, S;Bernkop-Schnürch, A
通过制备碳酸单叔丁酯和碳酸二叔丁酯胰岛素衍生物，将可用的蛋白质氨基与活化的750 Da PEG反应，最后进行氨基脱保护，得到胰岛素-单甲氧基聚乙二醇衍生物。该程序可以获得高产率的胰岛素-1PEG 和胰岛素-2PEG。通过皮下注射到糖尿病小鼠体内进行的体内研究表明，这两种生物缀合物保持了天然的生物活性。在体外，聚乙二醇化被发现可以增强激素对蛋白酶的稳定性。与弹性蛋白酶一起孵育 1 小时后，天然胰岛素、胰岛素-1PEG 和胰岛素-2PEG 分别经历约 70%、30% 和 10% 的降解，而在胃蛋白酶存在下分别经历 100%、70% 和 50% 的降解。低分子量 PEG 的附着并没有显着（P > 0.05）改变胰岛素穿过肠粘膜的渗透行为。胰岛素-1PEG被配制成由硫醇化聚合物聚(丙烯酸)-半胱氨酸构成的粘膜粘附片。治疗剂在 5 小时内从这些片剂中持续释放。在体内，通过给糖尿病小鼠口服给药，从给药后第三小时开始，血糖水平下降了约40%，并且生物活性维持长达30小时。根据这些结果，聚乙二醇化胰岛素与用作药物载体基质的硫醇化聚合物的组合可能是口服胰岛素给药的有前途的策略。 (C) 2004 Elsevier B.V. 保留所有权利。
Insulin-monomethoxypoly(ethylene glycol) derivatives were obtained by preparation of mono- and di-terbutyl carbonate insulin derivatives, reaction of available protein amino groups with activated 750 Da PEG and, finally, amino group de-protection. This procedure allowed for obtaining high yield of insulin-1PEG and insulin-2PEG. In vivo studies carried out by subcutaneous injection into diabetic mice demonstrated that the two bioconjugates maintained the native biological activity. In vitro, PEGylation was found to enhance the hormone stability towards proteases. After 1 h incubation with elastase, native insulin, insulin-1PEG and insulin-2PEG undergo about 70, 30 and 10% degradation, respectively, while in the presence of pepsin protein degradation was 100, 70 and 50%, respectively. The attachment of low molecular weight PEG did not significantly (P > 0.05) alter insulin permeation behavior across the intestinal mucosa. lnsulin-1PEG was formulated into mucoadhesive tablets constituted by the thiolated polymer poly(acrylic acid)-cysteine. The therapeutic agent was sustained released from these tablets within 5 h. In vivo, by oral administration to diabetic mice, the glucose levels were found to decrease of about 40% since the third hour from administration and the biological activity was maintained up to 30 h. According to these results, the combination of PEGylated insulin with a thiolated polymer used as drug carrier matrix might be a promising strategy for oral insulin administration. (C) 2004 Elsevier B.V. All rights reserved.