MULTIMECHANISM BASED MODEL FOR ANTICANCER DRUGS

基于多机制的抗癌药物模型

• 批准号: 2517584
• 负责人: ROBERT M SNAPKA
• 金额: $ 22.92万
• 依托单位: OHIO STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1995
• 资助国家: 美国
• 起止时间: 1995-09-01 至 2000-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2517584
• 关键词: DNA replication; DNA topoisomerases; alternatives to animals in research; antineoplastics; chemical structure; cytotoxicity; drug screening /evaluation; enzyme inhibitors; plant extracts; simian virus 40; DNA replication; DNA topoisomerases; alternatives to animals in research; antineoplastics; chemical structure; cytotoxicity; drug screening /evaluation; enzyme inhibitors; plant extracts; simian virus 40

The hypothesis for the proposed study is that simian virus 40 (SV40) DNA replication can be used as a multi-mechanism-based assay to discover and purify new anticancer drugs from natural sources. This approach has a number of unique advantages. The strengths of the SV40 assay are: (1) It is very sensitive to the four major groups of anticancer drugs which disrupt DNA replication in mammalian cells (topoisomerase I inhibitors, topoisomerase II inhibitors, alkylating agents and DNA nicking agents). (2) It is unaffected by a host of cytotoxic compounds which have no anticancer activity. Many of these interfere with other drug discovery approaches such as cytotoxicity assays or mechanism-based assays using purified enzymes and receptors. (3) It provides specific mechanistic information at the crude extract stage. This valuable information is used to prioritize the activities for purification. (4) It has the potential to discover antineoplastic drugs with novel targets such as helicases, primases and other components of the DNA replication fork. (5) It is economical, since it requires neither purified enzymes nor substrates. The immediate Goal of the study is to discover, purify and characterize new anticancer drugs from natural sources. The SV40 system will be used to identify potential anticancer drugs , prioritize them and guide their purification. This will demonstrate the unique advantages of this approach to drug discovery. The molecular structures of the purified compounds will be determined, their effects on purified enzymes of DNA replication will be studied, and their cytotoxicity profiles against cultured human tumor cells will be established in order to determine which are most promising for continued development. The long-term goal is to demonstrate the advantages of the multi-mechanism approach, which may be adaptable to signal transduction pathways, cell cycle checkpoints and other target groups in cancer cells. The Specific Aims are: (I) to purify and characterize the topoisomerase II inhibitors already detected in Pachycormus discolor, Desmos cochinchiensis and Dasymaschalon trichophorum. Molecular structures will be obtained and compared to those of known topoisomerase II inhibitors. Enzymology will be done with purified topoisomerase II and cytotoxicity profiles against the human tumor cell panel will be obtained. (II) to use the SV40 assay to detect, prioritize and guide the purification of new antineoplastic drugs targeting mammalian DNA replication. The plants surveyed will be from the OSU collection of over 1,700 species and the NCI Natural Products Repository. The plant survey will take place in two stages: a preliminary survey of approximately 360 plants which will identify the plant families and genera best suited for more thorough sampling and a secondary, focused survey based on activity patterns detected in the preliminary survey. (III) to purify the potent DNA replication fork-arresting activities from Pachycormus discolor and Polyalthia cerasoides, and determine their mechanisms of action.

拟议的研究的假设是Simian病毒40（SV40）DNA 复制可以用作基于多机械的测定法，以发现和 从天然来源净化新的抗癌药物。这种方法有一个 唯一优势的数量。 SV40分析的优势是：（1） 对四个主要的抗癌药物非常敏感，这些药物 破坏哺乳动物细胞中的DNA复制（拓扑异构酶I抑制剂， 拓扑异构酶II抑制剂，烷基化剂和DNA入口剂）。 （2）它不受许多没有的细胞毒性化合物的影响 抗癌活性。其中许多干扰了其他药物发现 诸如使用细胞毒性测定或基于机制的测定方法的方法 纯化的酶和受体。 （3）它提供了特定的机械 在粗提取阶段的信息。使用这些宝贵信息 优先考虑净化活动。 （4）它具有潜力 为了发现具有新型靶标（例如解旋酶）的抗塑性药物， DNA复制叉的原始成分和其他成分。 （5）是 经济性，因为它既不需要纯化的酶也不需要底物。 研究的直接目标是发现，净化和表征 来自天然来源的新抗癌药。 SV40系统将使用 确定潜在的抗癌药物，优先考虑并指导他们 纯化。这将证明这一点的独特优势 药物发现方法。纯化的分子结构 将确定化合物，它们对DNA纯化酶的影响 将研究复制，并针对其细胞毒性曲线 将建立培养的人类肿瘤细胞以确定 这是持续发展最有希望的。长期目标 是为了证明多种力学方法的优势， 可能适应信号转导途径，细胞周期检查点 和癌细胞中的其他目标群。 具体目的是：（i）净化和表征拓扑异构酶 II在Pachycormus中已经检测到的II抑制剂 变色，desmos cochinchiensis和dasymaschalon trichophorum。分子 将获得结构，并将其与已知拓扑异构酶的结构进行比较 II抑制剂。 酶学将使用纯化的拓扑异构酶II完成 针对人类肿瘤细胞面板的细胞毒性特征将是 获得。 （ii）使用SV40测定法检测，优先级和指导 靶向哺乳动物DNA的新抗肿瘤药物的纯化 复制。调查的工厂将来自OSU的收藏 1,700种和NCI天然产品存储库。工厂调查 将在两个阶段进行：大约360的初步调查 可以识别最适合植物家庭和属的植物 基于活动的更彻底的采样和次要的，重点的调查 在初步调查中检测到的模式。 （iii）净化有效性 DNA复制的叉式截止性活动，来自pachycormus cloror和 多多硫硫代菌，并确定其作用机理。