MULTIMECHANISM BASED MODEL FOR ANTICANCER DRUGS

基于多机制的抗癌药物模型

• 批准号: 2101686
• 负责人: ROBERT M SNAPKA
• 金额: $ 19.53万
• 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 1998-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2101686
• 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

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.

本研究的假设是猿猴病毒 40 (SV40) DNA 复制可以用作基于多机制的测定来发现和 从天然来源中纯化新的抗癌药物。这种方法有一个 等多项独特优势。 SV40 检测的优点是： (1) 对四大类抗癌药物非常敏感 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、烷化剂和 DNA 切口剂）。 （2）它不受许多细胞毒性化合物的影响，这些化合物没有 抗癌活性。其中许多会干扰其他药物的发现 诸如细胞毒性测定或基于机制的测定等方法 纯化的酶和受体。 (3) 提供了具体的机制 粗提物阶段的信息。这些有价值的信息被使用 确定净化活动的优先顺序。 （4）有潜力 发现具有解旋酶等新靶点的抗肿瘤药物， 引物酶和 DNA 复制叉的其他成分。 (5) 是 经济，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、纯化和表征 来自天然来源的新抗癌药物。将使用SV40系统 识别潜在的抗癌药物，确定其优先顺序并指导其使用 纯化。这将体现出该产品的独特优势 药物发现的方法。纯化后的分子结构 将确定化合物及其对 DNA 纯化酶的影响 将研究复制，以及它们的细胞毒性特征 将建立培养的人类肿瘤细胞以确定 其中最有希望持续发展。长期目标 是为了展示多机制方法的优势， 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体。 具体目标是：（I）纯化和表征拓扑异构酶 已在厚球菌中检测到 II 抑制剂 discolor、Desmos cochinchiensis 和 Dasymaschalon trichophorum。分子 将获得结构并与已知拓扑异构酶的结构进行比较 II 抑制剂。 将使用纯化的拓扑异构酶 II 进行酶学分析 针对人类肿瘤细胞组的细胞毒性特征将是 获得。 (II) 使用 SV40 检测来检测、优先排序和指导 针对哺乳动物DNA的新型抗肿瘤药物的纯化 复制。所调查的植物将来自 OSU 收集的超过 1,700 个物种和 NCI 天然产物存储库。植物调查 将分两个阶段进行：对约 360 人进行初步调查 植物将识别最适合的植物科和属 更彻底的抽样和基于活动的二次、有针对性的调查 初步调查中发现的模式。 （三）净化强效 Pachycormus discolor 和 DNA 复制叉阻滞活性 Polyalthia cerasoides，并确定其作用机制。