APPLICATIONS OF MASS SPECTROMETRY IN PHARMACOLOGY

质谱在药理学中的应用

• 批准号: 2173680
• 负责人: Catherine C Fenselau
• 金额: $ 18.13万
• 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/2173680
• 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.

将解决药理学方面的几个问题，其中 可以通过确定的主要序列来取得重大进展 蛋白质或改变的蛋白质。 质谱的速度和准确性 将与化学和化学问题一起解决这些问题 酶促技术。 将开发新的质谱技术 支持蛋白质结构研究。 我们将继续研究旨在了解倍数 在化学疗法期间经常获得耐药性的机制， 表征两个诱导的共价隔离 金属硫蛋白和诱导谷胱甘肽转移酶。 烷基化 纯化的蛋白质将以三种治疗性芥末的特征， 具有一系列化学反应率。 另外， 通过激活的葡萄糖醛酸在人血清白蛋白和上的共价键合 将定义胞质谷胱甘肽-S-转移酶。 识别 修改后的残留物可能会为这些绑定站点提供新的信息 广泛发生的代谢产物，并提供了分子机制 特发性免疫原性与非甾体类抗炎有关 和其他药物。 聚乙烯乙二醇衍生化的腺苷脱氨酶（PEG-ADA）为 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一非常复杂的 混合物，以了解其血浆稳定性的结构决定因素 和免疫相容性，并实现新陈代谢研究。 这些将是 适用于正在开发的其他毒素的蛋白质治疗 用途。 结合这些研究将提供有关是否是否 血清和胞质蛋白的异种烷基化是随机或有序的， 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 这些对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 用碰撞激活或反应引起的电喷雾电离 解离。