Ultrahigh-throughput Identification of Molecular Targets of Natural Products

天然产物分子靶标的超高通量鉴定

• 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
• 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

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）：天然产品是潜在药物的丰富来源。为了加速天然产物的药物发现，必须确定其分子靶标，并从机械上阐明其生物利用度，毒性和治疗作用。但是，该领域仍然存在一些挑战。其中，目前尚无技术平台可以使用通常在定量高通量筛选（QHTS）中遇到的少量样品进行机械研究。为了响应RFA- AT-14-001，Newomics Inc.提议开发一个用于高通量和高含量生物测定天然产品的多摩s平台。核心技术将基于Newomics的突破性硅 - 微流体芯片，多键嘴发射器阵列芯片（MEA CHIP-2012 R＆D100奖），可实现液相色谱 - 液相色谱式质量分布式电离质谱（LC-NANOESI/MS）的高度，高度敏感的，高度且高度敏感的，高度且高度敏感的，高度且高度敏感的，高度及分析物（肽，蛋白质和代谢产物）在OMICS水平上，来自少量样品。在这个项目中，我们将在单个MEA芯片上以高通量和多路复用格式结合多种OMICS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。我们将在第一阶段中展示我们的多功能平台的原理化证明应用。 AIM 1，我们将开发ESI-MS测定法，以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物质的关键载体与选定数量的天然产物之间的相互作用。在AIM 2中，我们将开发基于OMICS的测定法，以探测HSA在全血和天然产品库中具有不同复杂性的相互作用。一旦开发，Newomics的MEA芯片可能会成为对血液或其他组织中天然产物分子靶标的高吞吐量鉴定的通用平台，并为药物发现和发育奠定了新的基础。