Protein Aggregation in Amorphous Solids

无定形固体中的蛋白质聚集

• 批准号: 8042629
• 负责人: Elizabeth M. Topp
• 金额: $ 28.21万
• 依托单位: PURDUE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-03-01 至 2013-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8042629
• 关键词: Address; Adverse effects; Amino Acid Sequence; Anions; Autoimmune Diseases; Cardiovascular Diseases; Chemicals; Communicable Diseases; Computer Simulation; Computing Methodologies; Data; Deuterium; Disease; Disulfides; Drug Formulations; Drug Industry; Electrospray Ionization; Ensure; Environment; Excipients; Glass; Goals; Health; Hot Spot; Hydrogen; Investments; Knowledge; Life; Malignant Neoplasms; Marketing; Measures; Methods; Mission; Modeling; Pathway interactions; Patients; Peptide Sequence Determination; Peptides; Pharmaceutical Preparations; Process; Property; Protein Conformation; Proteins; Reaction; Research; Resolution; Role; Route; Safety; Set protein; Site; Solid; Solutions; Somatropin; Structural Protein; Structure; Sulfhydryl Compounds; Testing; Time; Transition Elements; Transition Temperature; United States National Institutes of Health; Work; amorphous solid; base; commercialization; computerized tools; cost; design; drug development; immunogenic; improved; molecular dynamics; prevent; programs; protein aggregation; protein structure; reaction rate; research study; response; solid state; tool

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.

描述（由申请人提供）：蛋白质药物是制药行业增长最快的细分市场之一。对这些药物的强烈需求反映了它们治疗以前棘手的疾病的能力，包括癌症，传染病，自身免疫性疾病和心血管疾病。超过40％的当前销售蛋白质药品是无定形固体，这种形式通常被选为延长保质期并保持效力。然而，蛋白质药物在固态下经历了各种物理和化学降解过程。聚集是这些过程中最常见的过程之一。由于聚集体的存在与效力降低有关，并且与威胁生命的免疫副作用的潜力增加有关，因此必须在制造和存储期间检测和去除它们。这增加了产生蛋白质药物的成本，最终增加了患者的成本，并排除了无法有效稳定的有希望的新蛋白质药物的商业化。该研究计划的目的是基于对涉及的化学物质（即共价）和物理（即非共价）机制的透彻理解，以开发用于预防固态蛋白质聚集的合理方法。中心假设是，无定形固体中的蛋白质聚集是特定的共价反应和/或在蛋白质序列中暴露于聚集的“热点”的结果，可以通过设计固体环境来预防蛋白质序列。提出的针对特定目标1的研究将阐明在无定形固体中二硫化物交换和二硫化物的机制，并将识别控制这些反应的固体特性。研究检验了以下假设：这些共价聚集的共同途径有利于溶液和固态中的不同途径，并且受固体组成的影响。特定的目标2将使用氢/氘（H/D）交换和分子动力学模拟（MDS）来鉴定无代固体中非共价蛋白聚集的“热点”。该工作检验了以下假设：这些定量的高分辨率测量无定形固体中的蛋白质结构将与长期存储期间的非共价聚集有关。特定目标3将开发一个计算模型，该模型基于蛋白质和固体的特性来预测无定形固体中的蛋白质聚集，从而产生一种用于制定设计的工具，并识别对防止聚集至关重要的变量。这项工作与NIH的使命是促进该国保护和改善健康能力的使命，因为它解决了维护快速增长类药物的效力和安全的方法。这项工作还与该机构的目标是一致的，即通过提供将活跃蛋白质开发到可销售的药品的工具和知识来确保公共研究的持续高回报。当药物对患者施用时，蛋白质药物中骨料的存在增加了威胁生命的免疫原性反应的潜力。了解无定形固体中的骨料形成将有助于确保这种快速增长的药物类别的安全。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。