Insulin Formulations of Enhanced Stability

描述(申请人提供)：胰岛素制剂化学代表蛋白质科学和应用生物技术的十字路口。根据美国国立卫生研究院路线图的双重观点，我们建议利用蛋白质工程的一般原理来提高胰岛素制剂的稳定性。因此，这项应用寻求对蛋白质折叠和展开的基本见解的临床翻译。目前的胰岛素制剂必须储存在冰箱中，28天后丢弃。此外，患者被敦促监测小瓶的变色、玻璃磨砂或沉淀：蛋白质化学或物理降解的迹象。最令人担忧的是胰岛素纤颤，这是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，以保护患者免受不慎注射胰岛素纤维的影响。我们寻求开发新的胰岛素类似物和配方，即使在高温下长时间储存也是高度活性的，但抗纤颤。基于对胰岛素淀粉样变性中间体的初步研究，提出了两种新的设计思想：通过D-氨基酸手性稳定B链和在A链上引入辅助的锌结合位点。将对这些胰岛素类似物进行热力学、动力学和结构研究，以验证设计目标。 拟议的研究如果成功，将代表着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。延长稳定性和保质期的胰岛素制剂在冷藏不稳定的发展中国家将具有特别的优势。我们的模拟设计策略虽然侧重于胰岛素的结构特征，但在蛋白质科学中使用了广泛应用的概念和原理。因此，我们的策略可能会推广到后基因组时代，以提高不同蛋白质药物的效用。

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.