APPLICATIONS OF MASS SPECTROMETRY IN PHARMACOLOGY

质谱在药理学中的应用

• 批准号: 2173679
• 负责人: Catherine C Fenselau
• 金额: $ 20.35万
• 依托单位: UNIVERSITY OF MARYLAND BALTIMORE
• 依托单位国家: 美国
• 财政年份: 1987
• 资助国家: 美国
• 起止时间: 1987-07-01 至 1996-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2173679
• 关键词: active sites; adenosine deaminase; alkylation; chemical binding; chemical bond; glutathione transferase; high performance liquid chromatography; mass spectrometry; metallothionein; molecular weight; orphan disease /drug; pharmacology; protein denaturation; protein sequence; protein structure

Several problems in pharmacology will be addressed, in which significant progress can be made by determining the primary sequence of proteins or altered proteins. The speed and accuracy of mass spectrometry will be brought to these problems, in conjunction with chemical and enzymatic techniques. New mass spectrometry techniques will be developed in support of protein structure studies. We will continue studies directed at understanding the multiple mechanisms for the drug resistance often acquired during chemotherapy, by characterizing the covalent sequestration proposed for both induced metallothionein and induced glutathione-Stransferase. Alkylation of purified proteins will be characterized for three therapeutic mustards, which have a range of chemical reactivities. In addition, sites of covalent bonding by activated glucuronides on human serum albumin and on cytosolic glutathione-S-transferases will be defined. Identification of modified residues may provide new information about binding sites for these widely occurring metabolites, and provide a molecular mechanism for the idiopathic immunogenicity associated with nonsteroidal anti-inflammatory and other drugs. Polyethylene glycol-derivatized adenosine deaminase (PEG-ADA) is an orphan drug used in treatment of severe immunosuppression syndrome. Analytical methods will be developed to characterize this very complex mixture, to understand the structural determinants of its plasma stability and immunocompatibility, and to enable metabolism studies. These will be applicable to other PEGated proteins under development for therapeutic uses. In combination these studies will provide insight as to whether xenobiotic alkylation of serum and cytosolic proteins is random or ordered, and how structural features in the drug and protein influence selectivity. Mass spectrometry techniques under development in support of these studies of pharmacologically relevant proteins include reaction induced dissociation in our high performance tandem mass spectrometer, and electrospray ionization with collisional activation or reaction induced dissociation.

将解决药理学中的几个问题，其中 通过确定一级序列可以取得重大进展 蛋白质或改变的蛋白质。 质谱分析的速度和准确性 结合化学和 酶技术。 将开发新的质谱技术 支持蛋白质结构研究。 我们将继续研究以了解多重 化疗过程中经常获得的耐药机制 表征了针对诱导的共价隔离所提出的 金属硫蛋白和诱导型谷胱甘肽-S转移酶。 烷基化 纯化的蛋白质将针对三种治疗芥末进行表征， 具有一系列化学反应活性。 此外，网站 活性葡萄糖苷酸与人血清白蛋白等的共价键合 将定义胞质谷胱甘肽-S-转移酶。 鉴定 修饰的残基可以提供有关这些结合位点的新信息 广泛存在的代谢物，并提供了分子机制 与非甾体抗炎药相关的特发性免疫原性 和其他药物。 聚乙二醇衍生的腺苷脱氨酶 (PEG-ADA) 一种用于治疗严重免疫抑制综合征的孤儿药。 将开发分析方法来表征这种非常复杂的现象 混合物，以了解其等离子体稳定性的结构决定因素 和免疫相容性，并能够进行代谢研究。 这些将是 适用于其他正在开发的治疗性聚乙二醇化蛋白 用途。 结合起来，这些研究将提供关于是否 血清和胞质蛋白的外源烷基化是随机的或有序的， 以及药物和蛋白质的结构特征如何影响选择性。 正在开发的质谱技术支持 这些药理学相关蛋白质的研究包括反应 在我们的高性能串联质谱仪中诱导解离，以及 碰撞激活或诱导反应的电喷雾电离 解离。