Anticancer Agents from Endophytic Fungi of Desert Plants

沙漠植物内生真菌的抗癌剂

• 批准号: 6724773
• 负责人: LESLIE GUNATILAKA
• 金额: $ 24.22万
• 依托单位: UNIVERSITY OF ARIZONA
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-04-01 至 2007-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6724773
• 关键词: Fungi; Gramineae; SCID mouse; angiogenesis inhibitors; antineoplastics; biological products; breast neoplasms; drug discovery /isolation; glioma; medicinal plants; neoplasm /cancer pharmacology; small cell lung cancer; tissue /cell culture

DESCRIPTION: (provided by applicant) The overall goal of this project is to discover novel anticancer drugs from endophytic fungi (fungi that live in the intercellular spaces) of desert plants with the broad long term objective of creating a library of this unexploited source of natural products for future drug discovery programs. The specific aims of the research to be undertaken will include sampling of 30 diverse Sonoran Desert medicinal plants and grasses for endophytic fungi; separating individual fungi and storing these to create a "library of endophytic fungi of Sonoran Desert plants "; culturing 500 endophytic fungi selected from this library on the basis of their diverse taxonomy; processing these fungal cultures and screening the resulting extracts for anticancer activity; fractionating selected bioactive extracts to isolate pure compounds responsible for their observed biological activity; determination of the structure of each biologically active compound; large-scale production of up to 3 compounds with unique structures and novel mechanisms of action followed by evaluation of their in vivo anticancer activity in human tumor xenograft mouse models. The methodologies to be utilized involve the separation and culturing of endophytic fungi in suitable solid or liquid media; successive extraction of fungal cultures with ethyl acetate and n-butanol and screening the resulting extracts in anticancer bioassays. Traditional proliferation/survival assays using three sentinel human cancer cell lines NCI-H460 (lung), and SF-268 (CNS-glioma)} and two new functional assays that monitor the inhibition of angiogenesis and the induction of heat shock protein expression will be used for screening. Dereplication of active extracts will be accomplished by taxonomically identifying the producing fungi based on literature reports and by analyzing bioactive extracts by LC-MS. Bioactivity-guided fractionation of unique active extracts will be performed using solvent-solvent partitioning, gel filtration and chromatography (CC, MPLC, prep. TLC, and HPLC). Structure elucidation of bioactive compounds will be achieved by a combination of spectroscopic (IR, UV, MS, and NMR) techniques, chemical manipulations, or X-ray crystallography as appropriate. Large-scale production (10-20 mg) of up to 3 selected compounds will be undertaken to obtain material for in vivo anticancer activity testing in hormone supplemented SCID mice bearing MCF-7 and T47D human tumor xenografts. We expect that having access to this unique source of bioactive natural products, combined with the use of both conventional and functional in vitro bioassays, followed by in vivo testing will lead to an efficient discovery of new anticancer agents which can be developed into clinically effective drugs with activity against solid tumors.

描述：（由申请人提供）该项目的总体目标是发现内生真菌的新型抗癌药物（生活在 沙漠植物的细胞间空间）具有广泛的长期目标 为未来的天然产品创建一个库的库 药物发现计划。进行研究的具体目的 将包括对30种不同的索诺兰沙漠药用植物和草的采样 用于内生真菌；分离单个真菌并存储这些真菌以创建一个 “索诺兰沙漠植物的内生真菌图书馆”；培养500 内生真菌是根据其多样的 分类学；处理这些真菌培养并筛选所得的提取物 用于抗癌活性；分馏所选的生物活性提取物以分离 纯化合物负责其观察到的生物学活性； 确定每个生物活性化合物的结构； 大规模生产多达3种具有独特结构和新颖的化合物 作用机理，然后评估其体内抗癌 人类肿瘤异种移植小鼠模型的活性。 要使用的方法涉及分离和培养 合适的固体或液体培养基中的内生真菌；连续提取 带有乙酸乙酯和N-丁醇的真菌培养物，并筛选由此产生的 抗癌生物测定中的提取物。传统的扩散/生存测定 使用三个哨兵人类癌细胞系NCI-H460（肺）和SF-268 （CNS-GLIOMA）}和两个新的功能测定法，以监测抑制 血管生成和热激蛋白表达的诱导将被使用 用于筛选。主动提取物的删除将通过 根据文献报告，从分类学识别生产真菌， 通过通过LC-MS分析生物活性提取物。生物活性引导的分馏 将使用溶剂 - 溶剂分区进行独特的活性提取物， 凝胶过滤和色谱法（CC，MPLC，Prep。TLC和HPLC）。结构 通过组合，将实现生物活性化合物的阐明 光谱学（IR，UV，MS和NMR）技术，化学操作或 X射线晶体学适当。大规模生产（10-20毫克） 将进行3种选定的化合物以获取体内材料 抗癌的抗癌活性测试补充了带有MCF-7和的SCID小鼠 T47D人类肿瘤异种移植物。 我们希望能够获得这种独特的生物活性自然来源 产品，结合使用常规和功能性的体外功能 生物测定，然后进行体内测试将导致有效发现 可以发展为临床有效药物的新抗癌剂 对实体瘤的活性。