Ultrahigh-throughput Identification of Molecular Targets of Natural Products

说明(申请人提供)：天然产物是潜在药物的丰富来源。为了加速从天然产物中发现药物，必须确定它们的分子靶点，并从机械上阐明它们的生物利用度、毒性和治疗效果。然而，该领域仍然存在几个挑战。其中，目前还没有一个技术平台可以使用定量高通量筛查(QHTS)中通常遇到的少量样本来进行从“Hits”到“Leads”的机械性研究。作为对RFA-AT-14-001的响应，Neomeics Inc.建议开发一个多组学平台，用于高通量和高含量的天然产品生物分析。核心技术将基于Neomeics的突破性硅微流控芯片-多喷嘴发射器阵列芯片(MEA芯片-2012 R&D100奖)，该芯片实现了基于液相色谱-纳米电喷雾电离质谱仪(LC-NanESI/MS)的、高灵敏度、高特异性、高通量和多路测量的多类别分析物(肽、蛋白质和代谢物)在OMICS水平上的小批量样品测量。在这个项目中，我们将以高通量和多路传输的形式，在单个MEA芯片上整合多种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。我们将演示我们的多组学平台在第一阶段的原则证明应用。 目的1、建立电喷雾质谱(ESI-MS)分析方法，筛选血浆中含量最丰富的蛋白质、内源性和外源性物质的关键载体--人血清白蛋白(HSA)与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OMICS的分析方法，以探索全血中HSA与不同复杂程度的天然产物库之间的相互作用。一旦开发出来，Neomeics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点的通用平台，并为药物发现和开发奠定新的基础。

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.