Protein Aggregation in Amorphous Solids

描述（由申请人提供）：蛋白质药物是制药行业增长最快的部分之一。对这些药物的强劲需求反映了它们治疗以前难治性疾病的能力，包括癌症，传染病，自身免疫性疾病和心血管疾病。目前市场上超过40%的蛋白质药物产品是无定形固体，通常选择这种形式来延长保质期并保持效力。然而，蛋白质药物在固态下经历各种物理和化学降解过程。聚合是这些过程中最常见的一种。由于聚集体的存在与效力降低和危及生命的免疫原性副作用的可能性增加有关，因此必须在生产和储存期间检测并去除它们。这增加了生产蛋白质药物的成本，最终增加了患者的成本，并阻碍了不能有效稳定的有前景的新蛋白质药物的商业化。这项研究计划的目标是开发合理的方法，以防止蛋白质聚集在固态的基础上彻底了解化学品（即，共价的）和物理的（即，非共价）机制。核心假设是蛋白质在无定形固体中的聚集是特定共价反应和/或蛋白质序列中易于聚集的“热点”暴露的结果，这两者都可以通过设计固体环境来防止。针对特定目标1拟定的研究将阐明无定形固体中硫醇-二硫键交换和二硫键混乱的机制，并将确定控制这些反应的固体性质。研究测试的假设，这些常见的共价聚集途径有利于不同的途径在溶液中和在固体状态，并受固体组成。具体目标2将使用氢/氘（H/D）交换和分子动力学模拟（MDS）确定非共价蛋白质聚集在无定形固体中的“热点”。这项工作测试的假设，这些定量的，高分辨率的措施，蛋白质结构的无定形固体将与非共价聚集在长期储存。具体目标3将开发一种计算模型，根据蛋白质和固体的性质预测蛋白质在无定形固体中的聚集，为配方设计和确定对防止聚集至关重要的变量提供工具。这项工作与NIH提高国家保护和改善健康能力的使命有关，因为它涉及保护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，即通过提供将活性蛋白质开发成可销售的药品的工具和知识，确保公共研究投资的持续高回报。蛋白质药物产品中聚集体的存在增加了向患者施用药物时威胁生命的免疫原性应答的可能性。了解无定形固体中的聚集体形成将有助于确保这一快速增长的药物类别的安全性。这项工作还将通过为蛋白质药物制剂提供合理的基础来帮助控制药物开发成本。

DESCRIPTION (provided by applicant): Protein drugs are one of the fastest growing segments of the pharmaceutical industry. The strong demand for these drugs reflects their ability to treat previously intractable diseases, including cancers, infectious disease, autoimmune disorders and cardiovascular disease. More than 40% of currently marketed protein drug products are amorphous solids, a form often chosen to prolong shelf-life and preserve potency. Nevertheless, protein drugs undergo a variety of physical and chemical degradation processes in the solid state. Aggregation is one of the most common of these processes. Since the presence of aggregates is associated with decreased potency and with an increased potential for life-threatening immunogenic side effects, they must be detected and removed during manufacturing and storage. This adds to the cost of producing protein drugs, ultimately increasing the cost to the patient and precluding the commercialization of promising new protein drugs that cannot be stabilized effectively. The goal of this research program is to develop rational methods for preventing protein aggregation in the solid state based on a thorough understanding of the chemical (i.e., covalent) and physical (i.e., non- covalent) mechanisms involved. The central hypothesis is that protein aggregation in amorphous solids is the result of specific covalent reactions and/or the exposure of aggregation-prone "hot spots" in the protein sequence, both of which can be prevented by designing the solid environment. Studies proposed for Specific Aim 1 will elucidate the mechanisms of thiol-disulfide exchange and disulfide scrambling in amorphous solids and will identify solid properties that control these reactions. The studies test the hypothesis that these common routes of covalent aggregation favor different pathways in solution and in the solid state and are influenced by solid composition. Specific Aim 2 will identify "hot spots" for non-covalent protein aggregation in amorphous solids using hydrogen/deuterium (H/D) exchange and molecular dynamics simulation (MDS). The work tests the hypothesis that these quantitative, high resolution measures of protein structure in amorphous solids will correlate with non-covalent aggregation during long-term storage. Specific Aim 3 will develop a computational model that predicts protein aggregation in amorphous solids based on properties of the protein and solid, producing a tool for formulation design and identifying variables critical to preventing aggregation. 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. The presence of aggregates in protein drug products increases the potential for life-threatening immunogenic responses when the drugs are administered to patients. Understanding aggregate formation in amorphous solids will help ensure the safety of this rapidly growing drug class. The work will also help to control drug development costs by providing a rational basis for protein drug formulation.