GOALI: Aggregation of Protein Therapeutics in Aqueous Solutions

目标：蛋白质治疗药物在水溶液中的聚集

• 批准号: 0138595
• 负责人: Theodore Randolph
• 金额: $ 43.15万
• 依托单位: University of Colorado at Boulder
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-07-01 至 2006-06-30
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=0138595&HistoricalAwards=false
• 关键词: GOALI; Aggregation; Protein; Therapeutics; Aqueous

Therapeutic proteins provide numerous unique treatments for human diseases (e.g., diabetes, and cancer). However, due to their delicate three-dimensional structures, proteins are very difficult to keep stable during the required 18-24 month shelf life of a pharmaceutical product. Furthermore, it is currently not possible to predict which solution conditions will result in sufficient stability. The Principal Investigators (PIs) propose to conduct fundamental studies of critical physical properties of proteins to develop methods for predicting optimal solution conditions for long-term stability. This multidisciplinary project combines efforts from Chemical Engineering and Pharmaceutical Sciences Departments at the University of Colorado with those of Amgen, an industrial partner. The PIs propose to determine the effects of critical solution conditions (pH, ionic strength and presence of stabilizing excipients) on rate and pathway for aggregation of several model proteins. Levels, and kinetics for formation and loss (where applicable) of soluble oligomeric species and insoluble precipitates will be measured, and structure of proteins in aggregates will be determined with infrared and derivative UV spectroscopies. To understand these effects, the PIs will determine the effects of pH, ionic strength and stabilizing excipients on protein structure and thermodynamic properties. Protein secondary and tertiary structures will be studied with circular dichroism, infrared, fluorescence and derivative-UV spectroscopies. The free energy of unfolding and hydrogen-deuterium exchange rates will be measured to test the hypothesis that increasing the thermodynamic stability of the native state and shifting the native state ensemble toward most compact species are important factors for inhibiting protein aggregation. The second osmotic virial coefficient will be measured by light scattering to assess the effects of solution conditions on the colloidal stability of the protein systems. High-pressure spectroscopic techniques will be used to complement these measurements. Finally, the effect of solution conditions on chemical stability (e.g., deamidation, oxidation and non-native disulfide formation) of the model proteins will be determined by the industrial collaborators at Amgen, and linkages between physical and chemical stability of the model proteins will be elucidated.

治疗蛋白为人类疾病（例如糖尿病和癌症）提供了许多独特的治疗方法。但是，由于其精致的三维结构，在所需的18-24个月保质期间，蛋白质很难保持稳定。此外，目前无法预测哪些解决方案条件将导致足够的稳定性。 首席研究者（PIS）建议对蛋白质的关键物理特性进行基础研究，以开发预测长期稳定性最佳解决方案条件的方法。这个多学科的项目结合了科罗拉多大学化学工程和制药科学系的努力与工业合作伙伴的安尔根（Amgen）的努力。 PI建议确定关键溶液条件（pH，离子强度和稳定赋形剂的存在）对几种模型蛋白聚集的速率和途径的影响。将测量可溶性寡聚物种和不溶性沉淀物的形成和损失的水平和动力学，将测量聚集体中蛋白质的结构，并通过红外和衍生的紫外线光谱法确定。 为了了解这些影响，PI将确定pH，离子强度和稳定赋形剂对蛋白质结构和热力学特性的影响。蛋白质二级结构和三级结构将通过圆形二色性，红外，荧光和衍生物-UV光谱研究进行研究。将测量展开和氢汇率的自由能，以检验以下假设：提高天然状态的热力学稳定性并将天然状态合奏转移到大多数紧凑型物种上是抑制蛋白质聚集的重要因素。第二种渗透病毒系数将通过光散射来测量，以评估溶液条件对蛋白质系统胶体稳定性的影响。高压光谱技术将用于补充这些测量值。最后，模型蛋白的溶液条件对化学稳定性（例如，脱氨酸，氧化和非本地二硫键形成）的影响将由Amgen的工业合作者确定，并且将阐明模型蛋白的物理和化学稳定性之间的联系。