Zhang Aihong, Lin Yin, Nong Shirly, Zhao Wei, Dong Mei
Department of Pharmaceutical, The Third Affiliated Hospital of Harbin Medical University Harbin Heilongjiang 150081 P. R. China
Department of Clinical Medicine,School of Basic Medical Sciences, Harbin Medical University Harbin Heilongjiang 150040 P. R. China.
RSC Adv. 2020 Jun 30;10(42):25013-25021. doi: 10.1039/d0ra01010c. eCollection 2020 Jun 29.
Using the commercially available antidiabetic drug exenatide (exendin-4) as a model peptide, we designed a novel exenatide derivative, termed LEX-1, comprising a 12-mer albumin-binding peptide, a protease-sensitive linker and a native exenatide. In addition, site-specific PEGylation was performed using LEX-1 as a lead sequence to generate four exenatide derivatives (LEX-2 to LEX-5). Moreover, we determined the optimal molecular weight of maleimide-derivatized PEG for the site-specific PEGylation of LEX-1 by an stability assay and an hypoglycemic efficacy test. As a result, LEX-3 (PEG10 kDa) exerted enhanced proteolytic stability, rational release rate of free exenatide and the best glucose-stabilizing capability compared with others. In addition, the prolonged hypoglycemic effects of LEX-1 and LEX-3 were demonstrated in type 2 diabetic mice by multiple OGTTs and a hypoglycemic duration test. Furthermore, a pharmacokinetic test was conducted using Sprague Dawley (SD) rats; LEX-3 (PEG10 kDa) showed the best circulating of ∼119.7 h for exenatide release from LEX-3, suggesting that LEX-3 has the potential to be developed into a once-weekly antidiabetic agent. The consecutive 8 week treatment of both LEX-1 and LEX-3 exhibited enhanced beneficial efficacies on body weight gain, cumulative food intake, % fat and hemoglobin A1c (HbA1c) reduction compared with exenatide treatment. Meanwhile, the chronic administration of LEX-1 and LEX-3 also effectively improved the blood biochemical indexes. Our results indicate the enhanced antidiabetic effects of LEX-1 and LEX-3, and our strategy of PEGylation and albumin conjugation can be applied to other bioactive agents.
我们以市售抗糖尿病药物艾塞那肽（exendin-4）作为模型肽，设计了一种新型艾塞那肽衍生物，称为LEX-1，它由一个12聚体白蛋白结合肽、一个蛋白酶敏感连接子和天然艾塞那肽组成。此外，以LEX-1为先导序列进行位点特异性聚乙二醇化，生成了四种艾塞那肽衍生物（LEX-2至LEX-5）。此外，我们通过稳定性测定和降血糖功效测试，确定了用于LEX-1位点特异性聚乙二醇化的马来酰亚胺衍生化聚乙二醇的最佳分子量。结果表明，与其他衍生物相比，LEX-3（PEG 10 kDa）具有更高的蛋白水解稳定性、游离艾塞那肽的合理释放速率以及最佳的血糖稳定能力。此外，通过多次口服葡萄糖耐量试验（OGTT）和降血糖持续时间测试，在2型糖尿病小鼠中证明了LEX-1和LEX-3具有延长的降血糖作用。此外，使用斯普拉格-道利（SD）大鼠进行了药代动力学测试；LEX-3（PEG 10 kDa）显示出从LEX-3释放艾塞那肽的最佳循环时间约为119.7小时，这表明LEX-3有潜力开发成一种每周一次的抗糖尿病药物。与艾塞那肽治疗相比，连续8周给予LEX-1和LEX-3对体重增加、累积食物摄入量、脂肪百分比和糖化血红蛋白（HbA1c）降低均表现出增强的有益效果。同时，长期给予LEX-1和LEX-3也有效改善了血液生化指标。我们的结果表明LEX-1和LEX-3具有增强的抗糖尿病作用，并且我们的聚乙二醇化和白蛋白缀合策略可应用于其他生物活性剂。
