Insulin Formulations of Enhanced Stability

描述（由申请人提供）：胰岛素制剂化学代表了蛋白质科学和应用生物技术之间的交叉道路。在雅阁的NIH路线图的双重观点，我们建议利用蛋白质工程的一般原则，以提高胰岛素制剂的稳定性。因此，本申请寻求将基本见解临床转化为蛋白质的折叠和解折叠。目前的胰岛素制剂必须储存在冰箱中，28天后丢弃。此外，敦促患者监测小瓶的变色、玻璃结霜或沉淀：蛋白质化学或物理降解的迹象。主要关注的是胰岛素纤维化，一种类似于淀粉样蛋白形成的错误折叠过程。美国食品和药物管理局制定了严格的标准，以保护患者免受无意中注射胰岛素纤维的影响。我们寻求开发新型胰岛素类似物和制剂，其具有高活性，但即使在高温下长时间储存后也能抵抗原纤化。基于对胰岛素淀粉样蛋白生成中间体的初步研究，提出了两种新的设计思路：通过D-氨基酸手性稳定B-链的转角和在A链中引入辅助锌结合位点。这些胰岛素类似物的热力学，动力学和结构研究将被追求，以验证设计目标。 这些研究如果成功，将代表结构原理在一个具有巨大医学和经济意义的问题上的令人兴奋的应用。稳定性和保存期延长的胰岛素制剂在发展中国家特别有利，因为发展中国家的冷藏条件并不稳定。我们的类似物设计策略，虽然侧重于胰岛素的结构特征，采用的概念和原则，广泛应用于蛋白质科学。因此，我们的策略可以推广到提高后基因组时代的各种蛋白质药物的效用。

DESCRIPTION (provided by applicant): Insulin formulation chemistry represents a cross roads between protein science and applied biotechnology. In accord with the dual perspective of the NIH Roadmap, we propose to exploit general principles of protein engineering to improve the stability of insulin formulations. This application thus seeks clinical translation of basic insights into the folding and unfolding of proteins. Present insulin formulations must be stored in refrigerator and discarded after 28 days. In addition, patients are urged to monitor vials for discoloration, frosting of the glass, or precipitation: signs of chemical or physical degradation of the protein. Of principal concern is insulin fibrillation, a misfolding process analogous to amyloid formation. The Food & Drug Administration has set stringent standards to protect patients from inadvertent injection of insulin fibrils. We seek to develop novel insulin analogs and formulations that are highly active but resistant to fibrillation even after prolonged storage at elevated temperatures. Two novel design ideas are proposed based on preliminary studies of an insulin amyloidogenic intermediate: chiral stabilization of a B-chain ¿-turn by D-amino acids and introduction of an ancillary zinc-binding site in the A chain. Thermodynamic, kinetic and structural studies of these insulin analogues will be pursued to validate the design goals. The proposed studies, if successful, would represent the exciting application of structural principles to a problem of enormous medical and economic significance. Insulin formulations of prolonged stability and extended shelf life would be of particular advantage in the developing world where refrigeration is not consistently available. Our analog design strategies, although focused on structural features of insulin, employ concepts and principles of broad application in protein science. Accordingly, our strategy may generalize to enhance the utility of diverse protein pharmaceuticals in the post-genomic era.