Development of Next Generation Stabilization Agents for Cell based Therapeutics

描述（由申请人提供）：我们最近证明，低熔点有机盐，称为离子液体，可以合理地设计，以稳定蛋白质对热变性通过调节溶剂的性质。通过明智地选择阳离子、阴离子和官能团，不仅可以调节稳定特性，而且还可以根据应用定制一系列物理性质，例如pH、粘度和盐的解离程度（即渗透压）。我们还能够通过合理选择阴离子来调节离子液体的细胞毒性。我们建议将这些令人兴奋的结果扩展到细胞应用，其中许多相同的功能是在细胞稳定制剂所需的。“任务特异性”分子设计规则的确定和应用将使保存科学超越现有材料的标准试错方法，朝着针对特定组织和储存要求合理设计细胞保护剂的方向发展。具体而言，我们力求：目标一：确定离子液体的结构特征和物理性质，从而稳定哺乳动物细胞膜的关联和渗透。目标二：确定生物相容性离子液体与水和糖基稳定分子（如海藻糖）的组合物的热物理行为和玻璃形成特性。目标3：使用三种不同的保存方法：缓慢冷却、快速冷却和脱水，确定保存在基于离子液体的保存溶液中的细胞的保存后活力和保质期。

DESCRIPTION (provided by applicant): We have recently demonstrated that low melting point organic salts, called ionic liquids, can be rationally designed to stabilize proteins against thermal denaturation by modulating solvent properties. By judicious choice of cation, anion, and functional groups, not only can the stabilization characteristics be tuned, but a range of physical properties, such as the pH, viscosity, and the extent of dissociation of the salt (i.e osmolality) can also be tailored to the application. We have also been able to modulate the cytotoxicity of ionic liquids by rational selection of anions. We propose to extend these exciting results to cellular applications, where many of the same features are desired in a cell stabilizing formulation. The determination and application of 'task-specific' molecular design rules will move preservation science beyond the standard trial and error approach with existing materials towards rationally designed cell protectants for specific tissue and storage requirements. Specifically we seek to: Aim #1: Determine the structural features and physical properties of ionic liquids that result in stabilizing associations and penetration of mammalian cell membranes. Aim #2: Determine the thermal-physical behavior and glass-forming characteristics of biocompatible ionic liquids in composition with water and sugar-based stabilizing molecules, such as trehalose. Aim #3: Establish the post preservation viability and shelf-life of cells stored in ionic liquid based preservation solutions using three different preservation methods: slow-cooling, rapid-cooling, and dehydration.