Injectable protease-operated depots of glucagon-like peptide-1 provide extended and tunable glucose control
Injectable protease-operated depots of glucagon-like peptide-1 provide extended and tunable glucose control
复制标题
DOI：
10.1073/pnas.1214518110
复制
发表时间：
2013-02-19
影响因子：
11.1
通讯作者：
Chilkoti, Ashutosh
中科院分区：
文献类型：
--
作者：
Amiram, Miriam;Luginbuhl, Kelli M.;Chilkoti, Ashutosh
肽类药物是一类令人兴奋的药物，越来越多地用于治疗各种疾病;然而，它们的主要缺点是半衰期短，这决定了多次和频繁的注射以及不期望的“峰谷”药代动力学特征，这可能导致不期望的副作用。合成的延长释放制剂可以提供生物活性天然肽的延长释放，但是它们的合成性质可能是生产和利用的障碍。受这些限制的影响，我们开发了一种新的完全基因编码的肽递送系统-蛋白酶操作的贮库（POD）-以提供肽药物从可注射的s.c.仓库。我们通过融合胰高血糖素样肽-1的蛋白酶可裂解低聚物（一种2型糖尿病药物）和一种热响应性、形成储库的弹性蛋白样多肽（在低于体温时发生热触发的逆相变），从而形成可注射储库，证明了POD的概念验证。我们构建了胰高血糖素样肽-1 POD的合成基因，并证明了它们在大肠杆菌中的高产量表达和我们以前开发的非色谱方案的简单纯化。值得注意的是，单次注射胰高血糖素样肽-1 POD能够降低小鼠的血糖水平长达5天，比注射天然肽药物长120倍。这些发现表明，POD为肽治疗剂的持续递送提供了合成肽封装方案的第一种遗传编码的替代方案。
Peptide drugs are an exciting class of pharmaceuticals increasingly used for the treatment of a variety of diseases; however, their main drawback is a short half-life, which dictates multiple and frequent injections and an undesirable "peak-and-valley" pharmacokinetic profile, which can cause undesirable side-effects. Synthetic prolonged release formulations can provide extended release of biologically active native peptide, but their synthetic nature can be an obstacle to production and utilization. Motivated by these limitations, we have developed a new and entirely genetically encoded peptide delivery system-Protease Operated Depots (PODs)-to provide sustained and tunable release of a peptide drug from an injectable s.c. depot. We demonstrate proof-of-concept of PODs, by fusion of protease cleavable oligomers of glucagon-like peptide-1, a type-2 diabetes drug, and a thermally responsive, depot-forming elastin-like-polypeptide that undergoes a thermally triggered inverse phase transition below body temperature, thereby forming an injectable depot. We constructed synthetic genes for glucagon-like peptide-1 PODs and demonstrated their high-yield expression in Escherichia coli and facile purification by a nonchromatographic scheme we had previously developed. Remarkably, a single injection of glucagon-like peptide-1 PODs was able to reduce blood glucose levels in mice for up to 5 d, 120 times longer than an injection of the native peptide drug. These findings demonstrate that PODs provide the first genetically encoded alternative to synthetic peptide encapsulation schemes for sustained delivery of peptide therapeutics.