In Vitro Screening of New Compounds and Analogs in Cell-Based Assays

基于细胞的检测中新化合物和类似物的体外筛选

• 批准号: 6934167
• 负责人: F. Michael Hoffmann
• 金额: $ 4.06万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-05-01 至 2010-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6934167
• 关键词: antineoplastics; biomedical facility; cooperative study; drug discovery /isolation; drug screening /evaluation; tissue /cell culture

The goal of this UW NCDDG will be to identify novel compounds that demonstrate promise as leads for new anti-cancer therapeutics in preclinical assays. The NCDDG Core B will be responsible for in vitro assays of the compounds produced in Projects 1 and 2 and evaluation of the potency and mechanism of those compounds. Data from Core B will be used by the PI and the steering committee to select: 1)analogs that should be dropped from further analysis, 2) analogs that require further in vitro biological assays and 3) analogs that should be used by Core C and Project 3 for in vivo efficacy testing in autochthonous mouse models of human cancer. In order to efficiently screen many hundreds of new compounds per year in a cost efficient manner, we propose to perform the Core B services at the Keck-UWCCC Small Molecule Screening Facility (http://hts.wisc.edu). The Screening Facility is an existing core service facility for the UW Comprehensive Cancer Center. This facility has the existing infrastructure (cell culture facility, automated liquid handling robots and plate readers), experienced staff and a track record of rapidly screening thousands of chemical compounds in a diverse array of cell-based and biochemical assays. Use of the existing facility will permit faster and cheaper analysis of the NCDDG candidate compounds compared to running those assays in an individual academic laboratory or establishing a new assay facility with NCDDG funds. The goal of Core B is to provide a filter for which compounds move to in vivo testing and not to establish in detail the mechanism of action of large numbers of compound analogs. The Core will use four cell-based assays in 96-well formats that each take advantage of the robotics and experience of the screening facility. Compound analogs will be tested for cytotoxicity in a panel of eight human cancer cell lines obtained from the NCI panel of 60 cell lines. To provide more mechanistic insights into compound function we will focus on two key cellular phenotypes, cell survival and cell migration. Compounds will be assayed for their ability to trigger apoptosis and for their ability to alter the key cell survival mediator Akt. A subset of compounds will also be assayed for their effect on tumor cell migration in vitro. All data will be subject to statistical analysis to ensure the quality of information that is obtained on the new compounds and to facilitate the most informed decisions about the future testing of any compound analog.

UW NCDDG的目标将是确定新的化合物，这些化合物有望成为新的 临床前试验中的抗癌治疗。NCDDG Core B将负责体外检测 项目1和2中产生的化合物及其效力和机理的评价 化合物。PI和指导委员会将使用来自核心B的数据来选择：1) 应从进一步的分析中删除，2)需要进一步的体外生物学检测的类似物和3) 应由Core C和Project 3用于在体小鼠体内疗效测试的类似物 人类癌症的模型。为了在每年的成本中有效地筛选数百种新化合物 以高效的方式，我们建议在Keck-UWCCC小分子筛查中执行Core B服务 设施(http://hts.wisc.edu).筛选设施是UW现有的核心服务设施 综合癌症中心。该设施拥有现有的基础设施(细胞培养设施，自动化 液体搬运机器人和平板阅读机)、经验丰富的工作人员和快速筛选的记录 在一系列不同的基于细胞和生物化学的分析中发现了数千种化合物。使用 现有设施将使NCDDG候选化合物的分析速度更快、成本更低 在一个单独的学术实验室运行这些检测，或者与NCDDG建立一个新的检测设备 资金。核心B的目标是提供一个过滤器，对哪些化合物移动到体内测试，而不是 详细建立了大量复合类似物的作用机制。核心将使用四个 96孔格式的基于细胞的分析，每个都利用了机器人技术和经验 筛查设施。化合物类似物将在八个人类癌细胞小组中进行细胞毒性测试 从60个细胞系的NCI面板中获得的细胞系。提供更多关于化合物的机械性见解 功能我们将关注两个关键的细胞表型，细胞存活和细胞迁移。化合物将是 检测它们触发细胞凋亡的能力和改变关键细胞生存介质Akt的能力。一个 还将测试化合物的亚组分对肿瘤细胞体外迁移的影响。所有数据都将 必须进行统计分析，以确保获得的关于新化合物和 以便于对任何复合模拟的未来测试做出最明智的决定。