Protein Aggregation in Amorphous Solids

无定形固体中的蛋白质聚集

• 批准号: 8811973
• 负责人: Elizabeth M. Topp
• 金额: $ 29.72万
• 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 2016-02-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8811973
• 关键词: Address; Adverse effects; Affect; Amino Acids; Antibodies; Award; Biological Products; Chemicals; Computer Assisted; Computer Simulation; Deuterium; Development; Disulfides; Drug Costs; Drug Design; Drug Formulations; Drug Industry; Ensure; Environment; Excipients; Exposure to; Freeze Drying; Goals; Grant; Health; Hydrogen; Immune response; Immunoglobulin G; Industry; Investments; Kinetics; Knowledge; Label; Lead; Life; Ligands; Maps; Marketing; Measures; Methods; Mission; Modeling; Modification; Molecular; Molecular Chaperones; Organized by Structure Protein; Patients; Peptides; Performance; Pharmaceutical Preparations; Powder dose form; Process; Property; Proteins; Quantitative Structure-Activity Relationship; Reaction; Research; Resolution; Route; Safety; Site; Solid; Solutions; Somatropin; Staphylococcal Protein A; Streptococcus IgG Fc-binding protein; Sulfhydryl Compounds; Techniques; Testing; Ultraviolet Rays; United States Food and Drug Administration; United States National Institutes of Health; Work; adduct; amorphous solid; analog; base; chemical reaction; commercialization; cost; design; disulfide bond; drug development; immunogenic; improved; liquid chromatography mass spectrometry; novel; prevent; programs; protein aggregation; protein folding; protein structure; screening; solid state; tool

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.

描述（由申请人提供）：蛋白质药物是生物制药行业中生长最快的领域。虽然通常以溶液的形式给药，但许多蛋白质药物作为无定形固体粉末出售，这种形式通常是为了保持稳定性和延长保质期。但是，蛋白质仍然可以在固态下降解。可以说，最严重的降解类型是聚集。聚集体的存在与药物效力的变化有关，这可能比无骨料形式更大或更少。聚集体还与患者不良免疫反应的潜力增加有关，这可能是威胁生命的。结果，必须在制造蛋白质药物期间检测和去除聚集体。这增加了他们的成本，这是最终由公众承担的负担。这项正在进行的研究计划的目的是开发一种有效的，设计的方法，以根据对所涉及机制的透彻理解来防止无定形固体中的蛋白质聚集。中心假设是，无定形固体中的蛋白质聚集是特定的化学反应和蛋白质结构变化的结果，蛋白质结构可以用高分辨率定义，并通过设计固体环境而预防。具体目标1将评估蛋白质结构对无定形固体中硫醇 - 二硫化物交换的影响，这是共价聚集的最常见途径之一。研究检验了硫醇 - 二硫化物交换的速率和机制受蛋白质结构的影响，溶液和无定形固体的差异。特定的目标2将开发固态光解标记（SSPLL）和固态氢氘交换（SSHDX），以绘制具有高分辨率的无定形固体中的蛋白质 - 矩阵相互作用。该研究检验了SSHDX和SSPLL比目前的蛋白质结构和固体特性的指标更好的指标。特定目标3将产生人工伴侣，以抑制IgG抗体的聚集。研究检验了含有人工伴侣的固体制剂显示出比具有共同添加剂的对照的假设更大的IgG聚集抑制作用。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。