DCTD Screening Technologies

该项目是癌症治疗和诊断部门内的发展治疗计划药物发现和开发计划的主要部分。重点是临床前药物发现和临床前药物开发。 体外癌细胞系筛选（IVCCLS）？在2007财政年度，细胞系筛选增加了第60个肿瘤细胞系，MDA-MB-468，用于评价天然和合成药物的抗肿瘤活性。该筛选包括代表各种癌症的细胞系，包括白血病、黑色素瘤以及肺癌、结肠癌、脑癌、卵巢癌、乳腺癌、前列腺癌和肾癌。目的是优先考虑对特定肿瘤细胞系显示选择性生长抑制或细胞杀伤的合成化合物或天然产物样品，以进行进一步评价。改变了初步筛选模式，以在筛选中结合针对所有60种细胞系的初始单次药物剂量筛选。在一次给药试验中显示活性的药物在标准试验中重新检测，该标准试验由使用60种人肿瘤细胞系的5种浓度试验组成。用于识别活性药物的标准是基于5剂筛选的历史数据制定的，旨在确立20 - 25%的目标通过率，并捕获之前在标准筛选中显示具有活性的大多数药物。单剂量筛选的概念是更快地识别活性药物并消除非活性化合物，以便资源可以集中在活性化合物上。60细胞系筛选的独特之处在于，给定化合物产生的60细胞系剂量反应的复杂性导致可用于模式识别算法的生物反应模式。使用这些程序，有可能为测试化合物分配一个假定的作用机制，或者确定响应模式是独特的，并且与NCI数据库中包含的任何标准原型化合物的响应模式不相似。此外，在表征细胞系中的各种细胞分子靶标之后，可以选择最可能与特定靶标相互作用的化合物。 体外分子靶点筛选计划（MTSP）-体外分子靶点筛选计划（MTSP）是一项专门服务，为DCTD/DTP抗癌药物发现计划提供直接支持。该实验室已经开发了无细胞和基于细胞的高通量（HTS）分子靶向检测的基础设施和专业知识。中期战略计划发展了工作人员的经验、仪器和信息技术基础设施，以便对大型化学品库（超过140 000种化合物）开展筛选活动。已经完成了几项HTS筛查活动，目前正在进行其他活动。此外，MTSP通过开发用感兴趣的分子靶基因转染人肿瘤细胞的新方法和优化重组蛋白的表达系统以用作HTS分子靶筛选的试剂，获得了增强的分子生物学经验。为了补充这些初级筛选，实验室开发了用于在各种HTS筛选中显示活性的化合物的二级测试的测定。

This project is a major part of the Developmental Therapeutics Program drug discovery and development program within the Division of Cancer Treatment and Diagnosis. The emphasis is on preclinical drug discovery and preclinical drug development. In Vitro Cancer Cell Line Screen (IVCCLS) ¿ During FY2007, the cell-line screen added a 60thth tumor cell line, MDA-MB-468, for evaluation of the antitumor activity of both natural and synthetic agents. The screen includes cell lines that represent a variety of cancers, including leukemia, melanoma, and cancers of the lung, colon, brain, ovary, breast, prostate, and kidney. The aim is to prioritize, for further evaluation, synthetic compounds or natural product samples showing selective growth inhibition or cell killing of particular tumor cell lines. The primary screening paradigm was changed to incorporate an initial single drug-dose screen against all 60 cell lines in the screen. Drugs that show activity in the one-dose assay are retested in the standard assay that consists of a 5-concentration test using the 60 human tumor cell lines. The criteria used to identify active agents was developed based on historical data from the 5-dose screen and developed to establish a targeted pass rate of 20¿25% and to capture the majority of drugs that have previously been shown to be active in the standard screen. The concept of the one-dose screen is to more rapidly identify active drugs and eliminate inactive compounds so resources can be focused on the active compounds. The 60-cell line screen is unique in that the complexity of a 60-cell line dose response produced by a given compound results in a biological response pattern that can be utilized in pattern recognition algorithms. Using these programs, it is possible to assign a putative mechanism of action to a test compound, or to determine that the response pattern is unique and not similar to that of any of the standard prototype compounds included in the NCI database. In addition, following characterization of various cellular molecular targets in the cell lines, it is possible to select compounds most likely to interact with a specific target. In Vitro Molecular Targets Screening Program (MTSP) - The In Vitro Molecular Target Screening Program (MTSP) is a dedicated service providing direct support to the DCTD/DTP anticancer drug discovery program. This laboratory has developed the infrastructure and expertise for the operation of cell-free and cell-based high-throughput (HTS) molecular target-based assays. MTSP has developed the staff experience, instrumentation, and information technology infrastructure to carry out screening campaigns with large chemical libraries (>140,000 compounds). Several HTS screening campaigns have been completed and others are ongoing at the present time. In addition, MTSP has gained enhanced molecular biology experience by developing new methods for the transfection of human tumor cells with molecular target genes of interest and by optimizing expression systems for recombinant proteins to be utilized as reagents for our HTS molecular target screens. To complement these primary screens, the laboratory has developed assays for secondary testing of compounds showing activity in various HTS screens.