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.

威斯康辛大学 NCDDG 的目标是确定新化合物，这些化合物有望作为新药物的先导化合物。 临床前试验中的抗癌治疗。 NCDDG 核心 B 将负责体外测定 项目 1 和 2 中生产的化合物以及对这些化合物的效力和机制的评估 化合物。 PI 和指导委员会将使用来自 Core B 的数据来选择： 1) 类似物 应从进一步分析中删除，2) 需要进一步体外生物学测定的类似物，以及 3) Core C 和 Project 3 应使用的类似物用于本土小鼠的体内功效测试 人类癌症模型。为了以较低的成本每年有效地筛选数百种新化合物 为了高效地进行，我们建议在 Keck-UWCCC 小分子筛选中执行核心 B 服务 设施（http://hts.wisc.edu）。筛选设施是华盛顿大学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、自动化设施） 液体处理机器人和读板机）、经验丰富的员工和快速筛选的记录 在各种基于细胞和生化检测中分析数千种化合物。使用 与现有设施相比，现有设施将能够更快、更便宜地分析 NCDDG 候选化合物 在单个学术实验室中运行这些检测或与 NCDDG 建立新的检测设施 资金。 Core B 的目标是提供一个过滤器，使化合物能够进入体内测试，而不是进入体内测试。 详细建立大量化合物类似物的作用机制。核心将使用四个 96 孔格式的基于细胞的测定，每种测定都利用了机器人技术和经验 筛选设施。将在一组八个人类癌细胞中测试化合物类似物的细胞毒性 从 60 个细胞系的 NCI 组中获得的细胞系。提供对化合物更多的机械见解 在功能方面，我们将重点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 分析了它们触发细胞凋亡的能力以及改变关键细胞存活介质 Akt 的能力。一个 还将分析部分化合物对体外肿瘤细胞迁移的影响。所有数据将 进行统计分析，以确保获得的新化合物信息的质量 促进对任何化合物类似物的未来测试做出最明智的决策。