Protein Stabilization in Ionic Liquids

描述(申请人提供)：以蛋白质为基础的治疗药物在控制和治疗疾病方面显示出显著的效果。与传统的小分子药物疗法不同，蛋白质药物开发的一个主要障碍是在整个加工过程中以及在使用结束时的储存期间保持蛋白质处于折叠状态的挑战。患者的依从性随着给药复杂性的增加而降低，因此注射用液体制剂比需要患者重建的冻干剂量更受欢迎。离子液体(ILS)是一类在蛋白质药物的液体制剂中具有相当大潜力的材料。离子液体是一种具有离子性质但在室温下呈液态的材料。虽然传统的ILS是由咪唑类化合物和高度氟化的阴离子合成的，但ILS也可以由自然界中存在的盐、糖、氨基酸和生物分子合成-其中许多已经被批准为赋形剂。离子液体的令人兴奋的特点是，由于它们由化学上不同的离子组成，氢键特性和水的混和性可以根据混合物中作用的性质和阴离子的性质而有效地调整到应用中。以下具体目标代表了确定潜在的ILS在液体配方中用作蛋白质稳定剂的关键步骤：目标1：评估一组合理设计的ILS的细胞毒性和水的混和性，以确定毒性最低、亲水性最强的化合物，以便继续研究。目的#2：从目标1中优先考虑的ILS的子集中确定对细胞色素C、溶菌酶和牛血清白蛋白增溶作用最强的ILS。目的#3：确定所选择的ILS是否能够在长期储存和短期物理和化学应激下保持液体制剂中的蛋白质结构和功能。更广泛的影响：拟议的工作非常符合美国国立卫生研究院的目标，因为它将促进我们对液体中蛋白质稳定的理解，因为它与药物配方有关。此外，成功实现拟议的目标将支持、促进和维持以蛋白质为基础的治疗人类疾病的经济配方方面的进展。这将对国家健康和福利问题产生直接和直接的影响。公共卫生相关性：以蛋白质为基础的治疗药物在控制和治愈疾病方面显示出显著的效果。与传统的小分子药物疗法不同，蛋白质药物开发的一个主要障碍是在整个加工过程中以及在使用结束时的储存期间保持蛋白质处于折叠状态的挑战。离子液体(ILS)是一类在蛋白质药物的液体制剂中具有相当大潜力的材料。该项目涉及新设计的生物相容离子液体的合成、表征和优化，以促进蛋白质的增溶和稳定，并最终稳定地长期储存和输送基于蛋白质的治疗药物。拟议的工作将促进我们对液体中蛋白质稳定的理解，因为它与药物配方有关。此外，成功实现拟议的目标将支持、促进和维持以蛋白质为基础的治疗人类疾病的经济配方方面的进展。这将对国家健康和福利问题产生直接和直接的影响。

DESCRIPTION (provided by applicant): Protein-based therapeutic drugs have demonstrated significant efficacy in controlling and curing disease. Unlike traditional small molecule-based drug therapies, a major hurdle in the development of protein drugs is the challenge of maintaining the protein in the folded state throughout processing and also during storage at the end point-of-use. Patient compliance decreases as the complexity of administration increases, consequently liquid formulation of parenterals is highly preferred over lyophilized dosages that require reconstitution by the patient. Ionic liquids (ILs) are a class of materials that have considerable potential to provide advances in liquid formulation of protein pharmaceuticals. ILs is materials that have ionic character yet are in liquid form at room temperature. Although traditional ILs are synthesized from imidazolium-based actions and highly fluorinated anions, ILs can also be formulated from salts, sugars, amino acids, and biomolecules that exist in nature - many of which have already been approved as excipients. The exciting feature of ILs is that, because they consist of chemically distinct ions, the hydrogen-bonding character and the water miscibility can effectively be `tuned' to the application depending on the nature of the action and anion in the mixture. The following specific aims represent the critical steps in identifying potential ILs for use as protein stabilizers in liquid formulations: Aim #1: To evaluate the cyto-toxicity and water miscibility of a panel of rationally designed ILs to identify the least toxic, most hydrophilic compounds for continued studies. Aim #2: To identify the most solubilizing ILs for cytochrome C, lysozyme, and bovine serum albumin from the subset of ILs prioritized in Aim #1. Aim #3: To determine if the selected ILs can preserve protein structure and function in liquid formulation over long periods of storage and also under short term physical and chemical stresses. Broader Impact: The proposed work fits well with the goals of the NIH because it will advance our understanding of protein stabilization in liquids as it pertains to drug formulation. Furthermore, successful achievement of the proposed goals will support, promote, and sustain advances in the economical formulation of protein-based therapeutics for the treatment of human disease. This will have a direct and immediate impact on national health and welfare issues. PUBLIC HEALTH RELEVANCE: Protein-based therapeutic drugs have demonstrated significant efficacy in controlling and curing disease. Unlike traditional small molecule-based drug therapies, a major hurdle in the development of protein drugs is the challenge of maintaining the protein in the folded state throughout processing and also during storage at the end point-of-use. Ionic liquids (ILs) are a class of materials that have considerable potential to provide advances in liquid formulation of protein pharmaceuticals. This project involves the synthesis, characterization, and optimization of newly designed biocompatible ionic liquids to facilitate solubilization and stabilization of proteins, and ultimately stable long-term storage and delivery of protein-based therapeutics. The proposed work will advance our understanding of protein stabilization in liquids as it pertains to drug formulation. Furthermore, successful achievement of the proposed goals will support, promote, and sustain advances in the economical formulation of protein-based therapeutics for the treatment of human disease. This will have a direct and immediate impact on national health and welfare issues.