High-Throughput Screening Platform for Cancer Drug Discovery

癌症药物发现的高通量筛选平台

• 批准号: 10432025
• 负责人: Laurie L. Parker
• 金额: $ 70.38万
• 依托单位: PALO ALTO RESEARCH CENTER
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-06-01 至 2024-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10432025
• 关键词: Advanced Development; Affinity; Antineoplastic Agents; Binding; Biochemical Reaction; Biological Assay; Biological Models; Bromodomain; Buffers; CREBBP gene; Calorimetry; Consumption; Data; Data Analyses; Detection; Enzymes; Goals; Gold; Grant; Histones; Hour; Individual; Label; Libraries; Ligands; Lighting; Liquid substance; Malignant Neoplasms; Measurement; Measures; Methods; Microfluidics; Monitor; Morphologic artifacts; Oils; Optics; Peptides; Performance; Phosphopeptides; Phosphotransferases; Positioning Attribute; Proteins; Reaction; Reagent; Research Personnel; Resolution; SYK gene; Sampling; Screening procedure; Specificity; System; Systems Development; Techniques; Technology; Temperature; Therapeutic; Thermodynamics; Time; Titrations; Validation; Water; aqueous; assay development; base; biophysical properties; cancer therapy; drug development; drug discovery; enthalpy; enzyme activity; enzyme substrate; experimental study; high throughput screening; high-throughput drug screening; histone acetyltransferase; improved; inhibitor; instrument; instrumentation; interest; liquid crystal; millisecond; particle; receptor; screening; small molecule; src Homology Region 2 Domain; stoichiometry; targeted treatment; therapeutic target; tool development

Project Summary Targeted therapies have shown great promise in the treatment of cancer. High-throughput screening (HTS) campaigns have often relied on assays using labeled ligands or enzyme substrates. Artifacts associated with labeling have led to the erroneous identification of compounds that act in on the labeled substrate rather than the intended target. A label-free, solution-based HTS method will facilitate identification of compounds acting specifically on the intended targets. The goals of this proposed project are to advance development and validate this calorimetry platform as an accessible approach for high throughput ITC analysis, demonstrating its advantages to study a variety of cancer therapeutic targets. To achieve this goal, we will improve mixing of reagents in droplets, multiplex temperature readout from individual droplets, automate fluidic handling and data analysis, and validate the applicability of the improved technology to study: 1) SH2 domain/phosphopeptide interactions, 2) spleen tyrosine kinase (Syk) enzyme activity, and 3) Bromodomain/acetylated histone peptide interactions, further applied in small molecule screening. The realization of the spectro-optical calorimetry technology platform will enable full thermodynamic characterization of binding and enzymatic reactions at the throughput of 1536-well plate assays but with significantly reduced assay development time and larger numbers of repeats and analyte concentrations. The combination of increased throughput and reduced sample consumption will change the way researchers view calorimetry: rather than being seen as a technique for a limited number of high-value measurements, it will be viewed as a primary screening method for cancer therapeutic targets.

项目摘要 靶向治疗在癌症治疗中显示出巨大的前景。高通量筛选（HTS） 活动通常依赖于使用标记的配体或酶底物的测定。伪影与 标记导致错误地鉴定作用于标记底物的化合物， 预定目标。一种无标记的、基于溶液的HTS方法将有助于鉴定起作用的化合物。 特别是在预定目标上。该拟议项目的目标是促进发展， 验证此量热平台作为高通量ITC分析的可访问方法，证明其 研究多种癌症治疗靶点的优势。 为了实现这一目标，我们将改进液滴中试剂的混合，从液滴中读取多路温度读数， 单个液滴，自动化流体处理和数据分析，并验证改进的 研究技术：1）SH 2结构域/磷酸肽相互作用，2）脾酪氨酸激酶（Syk）酶 活性，和3）溴结构域/乙酰化组蛋白肽相互作用，进一步应用于小分子 筛选 光谱-光学量热技术平台的实现将使全热力学 在1536孔板测定的通量下表征结合和酶促反应，但 显著减少了测定开发时间和更大的重复次数和分析物浓度。的 增加的通量和减少的样品消耗相结合，将改变研究人员看待 量热法：而不是被视为一种技术，为有限数量的高价值的测量，它将是 被视为癌症治疗靶点的主要筛选方法。