Protein Aggregation in Amorphous Solids

无定形固体中的蛋白质聚集

• 批准号: 9022483
• 负责人: Elizabeth M. Topp
• 金额: $ 29.66万
• 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 2018-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9022483
• 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 Industry; Ensure; Environment; Excipients; Exposure to; Formulation; 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; Patients; Peptides; Performance; Pharmaceutical Preparations; Powder dose form; Process; Property; Proteins; Quantitative Structure-Activity Relationship; Reaction; Research; Resolution; Route; Safety; Site; Solid; 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形式出售，这种形式通常被选择用于保持稳定性并延长保质期。然而，蛋白质仍然可以在固态下降解。可以说，最严重的退化类型是聚合。聚集体的存在与药物效力的变化相关，其可能大于或小于无聚集体形式。聚集体还与患者不良免疫反应的可能性增加相关，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，最终由公众承担的负担。这项正在进行的研究计划的目标是开发一种有效的，设计的方法，以防止蛋白质聚集在无定形固体的基础上所涉及的机制的透彻理解。核心假设是蛋白质在无定形固体中的聚集是特定化学反应和蛋白质结构变化的结果，这些变化可以用高分辨率来定义，并通过设计固体环境来防止。具体目标1将评估蛋白质结构对无定形固体中硫醇-二硫键交换的影响，这是共价聚集的最常见途径之一。这些研究验证了以下假设：巯基-二硫键交换的速率和机制受蛋白质结构的影响，并且在溶液和无定形固体中不同。具体目标2将开发固态光解标记（ssPLL）和固态氢氘交换（ssHDX），以高分辨率绘制无定形固体中的蛋白质-基质相互作用。研究测试的假设，ssHDX和ssPLL是更好的指标聚集倾向的无定形固体比目前的措施，蛋白质结构和固体性质。特异性目标3将产生抑制IgG抗体聚集的人工分子伴侣。这些研究检验了以下假设：含有人工伴侣的固体制剂比含有普通添加剂的对照显示出更大的IgG聚集抑制。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。

项目成果

Thiol-disulfide interchange in the tocinoic acid/glutathione system during freezing and drying.

在冷冻和干燥过程中，生育酸/谷胱甘肽系统中的硫醇-二硫化物交换。

• DOI: 10.1002/jps.22206
• 发表时间: 2010
• 期刊: Journal of pharmaceutical sciences
• 影响因子: 3.8
• 作者: Thing,Mette;Zhang,Jun;Laurence,Jennifer;Topp,ElizabethM
• 通讯作者: Topp,ElizabethM

Fibrillation of Human Calcitonin and its Analogs: Effects of Phosphorylation and Disulfide Reduction.

• DOI: 10.1016/j.bpj.2020.11.009
• 发表时间: 2020-11
• 期刊: Biophysical journal
• 影响因子: 3.4
• 作者: Harshil K Renawala;K. B. Chandrababu;E. Topp

Fibrillation of human insulin B-chain by pulsed hydrogen-deuterium exchange mass spectrometry.

通过脉冲氢-氘交换质谱法测定人胰岛素 B 链的颤动。

• DOI: 10.1016/j.bpj.2022.10.042
• 发表时间: 2022
• 期刊: Biophysical journal
• 影响因子: 3.4
• 作者: Renawala,HarshilK;Topp,ElizabethM
• 通讯作者: Topp,ElizabethM

Thiol-Disulfide Exchange in Human Growth Hormone.

• DOI: 10.1007/s11095-016-1879-3
• 发表时间: 2016-06
• 期刊: Pharmaceutical research
• 影响因子: 3.7
• 作者: Chandrasekhar S;Moorthy BS;Xie R;Topp EM
• 通讯作者: Topp EM

Photolytic Labeling To Quantify Peptide-Water Interactions in Lyophilized Solids.

光解标记定量冻干固体中肽-水相互作用。

• DOI: 10.1021/acs.molpharmaceut.8b01031
• 发表时间: 2019
• 期刊: Molecular pharmaceutics
• 影响因子: 4.9
• 作者: Chen,Yuan;Topp,ElizabethM
• 通讯作者: Topp,ElizabethM