Core--Biology

核心--生物学

• 批准号: 6938238
• 负责人: John M. Pezzuto
• 金额: $ 13.49万
• 依托单位: PURDUE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-04-01 至 2010-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6938238
• 关键词: biological products; biology; biomedical facility; biotechnology; cancer prevention; carcinogenesis inhibitor; chemoprevention; drug discovery /isolation; laboratory mouse; laboratory rat; mammary gland; neoplasm /cancer pharmacology; neoplastic transformation; organ culture

The overall goal of this program project is to identify and evaluate novel natural product cancer chemopreventive agents. In the discovery process, activity-guided fractionation is employed, utilizing short-term bioassays as a monitor. The first goal of this core is to provide bioassay support of physiological relevance. As validated previously, the current test battery is comprised of assays designed to monitor inhibition of carcinogenesis at the stages of initiation (e.g., induction of quinone reductase activity in cell culture), promotion (e.g., inhibition of cyclooxygenase activity), and progression (e.g., induction of cell differentiation). Mechanistic-based high throughput assay systems have recently been devised, such as a fluorometric assay for inhibition of aromatase, and stably transfected HepG2 cells with reporter genes for ARE, ERE, PPAR, and NF-kappaB. Test materials and assay procedures actually used for specific fractionations are keyed on secondary discriminations also provided by this core. Once lead materials are structurally characterized, additional studies are performed in this core to more fully characterize the potential of the test agent to succeed in clinical intervention trials. With semi-preparative quantities of active leads (0.05-1 g), preliminary mechanistic evaluations will be performed. In addition to in vitro methodologies, this will involve short-term animal studies designed to assess effects on biomarkers of carcinogenesis (e.g., induction of phase II enzymes). In addition, with relatively small quantities of test materials, inhibition of tumorigenesis will be determined in the two-stage mouse skin model. Finally, based on the overall profile of structure, activity, natural abundance, etc., the most promising test materials will be evaluated for chemopreventive activity in full-term models of carcinogenesis with laboratory animals. The specific animal model and administration protocol will be based on the overall profile of the agent. The full-term tumorigenesis systems to be employed include the two-stage mouse skin model, the carcinogen (DMBA or NMU)-induced rat mammary model, the PIN (prostatic intraepithelial neoplasia) prostate model with male Noble rats, and the benzo(a)pyrene-induced lung tumor model in strain A mice. Complete data sets will be prepared and transmitted to organizations capable of promoting more advanced stages of development, such as the National Cancer Institute.

本项目的总体目标是鉴定和评估新的天然产物癌症化学预防剂。在发现过程中，采用活性导向分馏，利用短期生物测定作为监测。本核心的第一个目标是提供生理相关性的生物测定支持。正如之前验证的那样，目前的测试电池包括旨在监测癌变的起始阶段（例如，诱导细胞培养中的醌还原酶活性），促进阶段（例如，抑制环氧化酶活性）和进展阶段（例如，诱导细胞分化）的抑制。最近设计了基于机械的高通量分析系统，例如芳香酶抑制的荧光测定，以及稳定转染带有ARE， ERE， PPAR和NF-kappaB报告基因的HepG2细胞。实际用于特定分馏的测试材料和分析程序以该核心提供的二级鉴别为关键。一旦铅材料的结构特征，在该核心进行进一步的研究，以更充分地表征试验剂在临床干预中取得成功的潜力