Ultrahigh-throughput Identification of Molecular Targets of Natural Products

天然产物分子靶标的超高通量鉴定

基本信息

批准号：
8712293
负责人：
DAOJING WANG
金额：
$ 22.5万
依托单位：
NEWOMICS, INC
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-06-01 至 2015-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8712293
关键词：
Address Artemisinins Award Binding Binding Sites Biochemistry Bioinformatics Biological Assay Biological Availability Biological Factors Biomedical Research Biophysics Blood Chemicals Complement 4b Complex Crude Extracts Development Fluorescence Foundations Human Incubated Infusion procedures Label Libraries Liquid Chromatography Malaria Malignant Neoplasms Mass Spectrum Analysis Measurement Microfluidics Molecular Target Nigella sativa Paclitaxel Peptides Performance Pharmaceutical Preparations Phase Plasma Proteins Proteomics Publications Reaction Relative (related person)Reproducibility Research Research Personnel Sampling Sensitivity and Specificity Serum Albumin Signal Transduction Silicon Small Business Innovation Research Grant Solvents Source Technology Therapeutic Effect Time Tissues Toxic effect Validation Variant Walkers Warfarin Whole Blood Work adduct artemisinine assay development base blood product cancer stem cell cost drug development drug discovery experience high throughput screening innovation instrument ionization metabolomics nano-electrospray protein complex protein metabolite public health relevance response small molecule stem cell biology stoichiometry

项目摘要

DESCRIPTION (provided by applicant): Natural products are rich sources for potential drugs. To accelerate drug discovery from natural products, it is imperative to identify their molecular targets, and mechanistically elucidate their bioavailability, toxicity, and therapeutic effects. However, there remain several challenges in the field. Among them, there is currently no technical platform that could perform mechanistic studies from the "hits" to "leads" using the small volumes of samples that are typically encountered in quantitative high-throughput screening (qHTS). In response to RFA- AT-14-001, Newomics Inc. proposes to develop a multi-omics platform for high-throughput and high-content bioassays of natural products. The core technology will be based on Newomics' breakthrough silicon- microfluidic-chip, the multinozzle emitter array chip (MEA chip-2012 R&D100 award), which enables liquid chromatography-nanoelectrospray ionization mass spectrometry (LC-nanoESI/MS)-based, highly sensitive, highly specific, high-throughput, and multiplex measurements of multiclass analytes (peptides, proteins, and metabolites) at the Omics level, from small volumes of samples. In this project, we will incorporate multiple Omics capabilities, including top-down proteomics, bottom-up proteomics, and metabolomics, on a single MEA chip, in a high-throughput and multiplex format. We will demonstrate proof-of-principle applications of our multi- omics platform in the Phase I. In Aim 1, we will develop the ESI-MS assays to screen the interactions between purified human serum albumin (HSA), the most abundant protein and the key carrier of endogenous and exogenous substances in the plasma, and a selected number of natural products. In Aim 2, we will develop the Omics-based assays for probing interactions between HSA in whole blood and libraries of natural products with different complexity. Once developed, Newomics' MEA chip may become a universal platform for high- throughput identification of molecular targets of natural products in the blood or other tissues, and lay a new foundation for drug discovery and development.

描述（由申请人提供）：天然产物是潜在药物的丰富来源。为了加速天然产物的药物发现，必须确定其分子靶点，并从机制上阐明其生物利用度、毒性和治疗效果。然而，该领域仍然存在一些挑战。其中，目前还没有技术平台可以使用定量高通量筛选（qHTS）中通常遇到的小体积样本来进行从“命中”到“先导”的机制研究。为了响应 RFA-AT-14-001，Newomics Inc. 提议开发一个多组学平台，用于天然产物的高通量和高内涵生物测定。核心技术将基于Newomics突破性的硅微流控芯片——多喷嘴发射器阵列芯片（MEA芯片-2012年R&D100奖），该芯片能够实现基于液相色谱-纳米电喷雾电离质谱（LC-nanoESI/MS）的、高度在组学水平上对多类分析物（肽、蛋白质和代谢物）进行灵敏、高度特异性、高通量和多重测量，小体积样品。在这个项目中，我们将以高通量和多重格式在单个 MEA 芯片上整合多种组学功能，包括自上而下的蛋白质组学、自下而上的蛋白质组学和代谢组学。我们将在第一阶段展示我们的多组学平台的原理验证应用。目标 1，我们将开发 ESI-MS 测定法来筛选纯化的人血清白蛋白 (HSA)（血浆中最丰富的蛋白质和内源性和外源性物质的关键载体）与选定的一些天然产物之间的相互作用。在目标 2 中，我们将开发基于组学的检测方法，用于探测全血中的 HSA 与具有不同复杂性的天然产物库之间的相互作用。一旦开发完成，Newomics的MEA芯片可能成为高通量鉴定血液或其他组织中天然产物分子靶标的通用平台，并为药物发现和开发奠定新的基础。