Activity-based Proteomics for Toxicological Analysis

描述(由申请人提供)： ActivX生物科学公司将开发和验证一种新的毒素蛋白质组学平台，将蛋白质活性的变化与化合物毒理学联系起来。该平台使用荧光化学探针来询问来自任何生物样本的超家族相关的催化活性蛋白。与更常用的转录谱方法相比，蛋白质活性的变化有望更好地解释毒物的分子机制，并与之相关。这项第二阶段的研究将展示这一蛋白质活性平台的实用性。这项研究分为几个阶段。1)该平台将被建立为一个预测化合物毒性的平台，通过调查a)细胞系和b)动物组织中暴露于化合物的蛋白质活性，这些化合物已经通过药理学、基因组学和其他技术进行了很好的研究。2)所得到的蛋白质活性图谱将被集成到数据库中，该数据库还包括公开可用的转录数据，并将允许处理与丝氨酸水解酶、半胱氨酸蛋白酶、激酶、环氧化物水解酶和谷胱甘肽-S转移酶家族的蛋白质活性水平的变化无缝关联。其中一些酶家族的基于活性的探针(ABP)是细胞穿透的，这是一个有用的功能，允许在体内而不是体外测量细胞系中的蛋白质活性。3) 将合成目前不是细胞渗透性的ABPs的细胞渗透性衍生物。4)为了扩大我们分析能力的广度，ActivX还将设计、合成和验证磷酸酶和细胞色素P450家族的ABP。这一平台将有助于检测包括治疗药物在内的化学品的毒性，并将对毒性的分子基础进行分类和探索。因此，这项技术在缩短化合物早期开发的时间和成本以及揭示不同化合物潜在的毒性机制方面具有巨大的潜力。ActivX计划将其技术商业化，建立一个毒素蛋白质组数据库，并为制药业提供服务，在药物筛选过程的早期检测化合物的毒性。

DESCRIPTION (provided by applicant): ActivX Biosciences (ActivX) will develop and validate a novel toxicoproteomics platform to correlate changes in protein activity with compound toxicology. This platform uses fluorescent chemical probes to interrogate catalytically active proteins related by super-family derived from any biological sample. Alterations in protein activity are expected to be more interpretable and relevant to toxicant molecular mechanism than the more commonly used transcript-profiling approaches. This Phase II study will demonstrate the utility of this protein activity platform. This study has several stages. 1) The platform will be established as a predictive platform for compound toxicity by investigating protein activities of a) cell lines and b) tissues from animals exposed to compounds that have been well-studied using pharmacology, genomics, and other techniques. 2) The resulting protein activity profiles will be integrated into a database that also includes publicly available transcription data, and will allow for seamless correlation of treatments with changes in protein activity levels of the serine hydrolase, cysteine protease, kinase, epoxide hydrolase, and glutathione-S-transferase families. The activity-based probes (ABPs) for some of these enzyme families are cell-permeable, which is a useful feature that allows for in vivo, rather than in vitro measurements of protein activity in cell lines. 3) Cell-permeable derivatives of ABPs that are currently not cell permeable will be synthesized. 4) To expand the breadth of our profiling abilities, ActivX will also design, synthesize and validate ABPs for the phosphatase and cytochrome P450 families. This platform will be useful for detecting toxicity of chemicals, including therapeutics, and will be important for classifying and exploring the molecular basis of toxicities. Therefore, this technology has a great potential for reducing the time and cost in early compound development, and for revealing underlying toxicity mechanisms of different compounds. ActivX plans to commercialize its technology by establishing a toxicoproteomic database, and by providing a service to the pharmaceutical industry that will detect toxicities of compounds early in the drug screening process.