Protein Aggregation in Amorphous Solids

描述（由申请人提供）：蛋白质药物是生物制药工业中增长最快的部门。虽然通常以溶液形式给药，但许多蛋白质药物以无定形固体粉末的形式出售，选择这种形式通常是为了保持稳定性和延长保质期。然而，蛋白质在固态下仍然可以降解。可以说，最严重的退化类型是聚集。聚集体的存在与药物效力的变化有关，其效力可能比无聚集体的形式更大或更小。聚集体还与患者发生不良免疫反应的可能性增加有关，这可能危及生命。因此，在蛋白质药物的生产过程中，必须检测和去除聚集体。这增加了他们的成本，最终由公众承担。这个正在进行的研究项目的目标是开发一种有效的，设计的方法来防止蛋白质聚集在无定形固体中，基于对所涉及的机制的透彻理解。核心假设是蛋白质在非晶固体中的聚集是特定化学反应和蛋白质结构变化的结果，这种变化可以用高分辨率来定义，并通过设计固体环境来防止。特异性目标1将评估蛋白质结构对无定形固体中硫醇-二硫交换的影响，这是共价聚集的最常见途径之一。该研究验证了蛋白质结构对硫醇-二硫交换速率和机制的影响，并在溶液和非晶固体中有所不同。Specific Aim 2将开发固态光解标记（ssPLL）和固态氢氘交换（ssHDX），以高分辨率绘制非晶态固体中蛋白质-基质相互作用的图谱。这些研究验证了一个假设，即ssHDX和ssPLL是非晶固体中聚集倾向的更好指标，而不是目前的蛋白质结构和固体性质指标。特异性Aim 3将产生人工伴侣，抑制IgG抗体的聚集。这些研究验证了一个假设，即含有人工伴侣的固体配方比普通添加剂的对照更能抑制IgG聚集。这项工作与NIH的使命有关，即提高国家保护和改善健康的能力，因为它解决了保持快速增长的一类药物的效力和安全性的方法。这项工作也符合该机构的目标，即通过为将活性蛋白开发成可销售的药物产品提供工具和知识，确保公共研究投资的持续高回报。

DESCRIPTION (provided by applicant): Protein drugs are the fastest growing sector of the biopharmaceutical industry. While usually administered as solutions, many protein drugs are sold as amorphous solid powders, a form often chosen to preserve stability and prolong shelf-life. Proteins can still degrade in the solid state, however. Arguably, the most serious type of degradation is aggregation. The presence of aggregates is associated with changes in drug potency, which may be either greater or less than in the aggregate-free form. Aggregates are also associated with an increased potential for adverse immune responses in patients, which can be life-threatening. As a result, aggregates must be detected and removed during the manufacture of protein drugs. This adds to their cost, a burden ultimately borne by the public. The goal of this ongoing research program is to develop an efficient, designed approach to preventing protein aggregation in amorphous solids based on a thorough understanding of the mechanisms involved. The central hypothesis is that protein aggregation in amorphous solids is the result of specific chemical reactions and changes in protein structure that can be defined with high resolution and prevented by designing the solid environment. Specific Aim 1 will assess the effects of protein structure on thiol-disulfide exchange in amorphous solids, one of the most common routes to covalent aggregation. The studies test the hypothesis that the rates and mechanisms of thiol-disulfide exchange are affected by protein structure and differ in solution and in amorphous solids. Specific Aim 2 will develop solid-state photolytic labeling (ssPLL) and solid- state hydrogen deuterium exchange (ssHDX) to map protein-matrix interactions in amorphous solids with high resolution. The studies test the hypothesis that ssHDX and ssPLL are better indicators of aggregation propensity in amorphous solids than current measures of protein structure and solid properties. Specific Aim 3 will create artificial chaperones that inhibit the aggregation of IgG antibodies. The studies test the hypothesis that solid formulations containing artificial chaperones show greater inhibition of IgG aggregation than controls with common additives. The work is relevant to the NIH mission of advancing the Nation's capacity to protect and improve health in that it addresses methods to preserve the potency and safety of a rapidly growing class of drugs. The work is also consistent with the agency's goal of ensuring a continued high return on the public investment in research by providing tools and knowledge for developing active proteins into marketable drug products.