Anticancer Agents from Endophytic Fungi of Desert Plants

沙漠植物内生真菌的抗癌剂

• 批准号: 6612792
• 负责人: LESLIE GUNATILAKA
• 金额: $ 23.92万
• 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/6612792
• 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）本项目的总体目标是 从内生真菌（生活在植物体内的真菌）中发现新的抗癌药物。 沙漠植物的广泛的长期目标是 为未来创建一个未开发的天然产品来源库 药物发现项目拟进行的研究的具体目标 将包括30种不同的索诺兰沙漠药用植物和草的采样 对于内生真菌;分离单个真菌并储存这些真菌以创建 “索诺兰沙漠植物内生真菌库“;培养500 根据其多样性从该文库中选择内生真菌 分类学;处理这些真菌培养物并筛选所得提取物 用于抗癌活性;分级选择的生物活性提取物以分离 纯化合物负责其观察到的生物活性; 确定每种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， 作用机制，随后评价其体内抗癌作用 在人肿瘤异种移植小鼠模型中的活性。 所使用的方法包括分离和培养 内生真菌在合适的固体或液体介质中;连续提取 用乙酸乙酯和正丁醇进行真菌培养，并筛选得到的 抗癌生物测定中的提取物。传统的增殖/存活测定 使用三种前哨人癌细胞系NCI-H460（肺）和SF-268 （CNS-胶质瘤）}和两种新的功能测定， 血管生成和热休克蛋白表达的诱导将被用于 用于筛选。活性提取物的去复制将通过 根据文献报道对产菌进行分类鉴定， 通过LC-MS分析生物活性提取物。 将使用溶剂-溶剂分配进行独特的活性提取， 凝胶过滤和色谱法（CC、MPLC、制备型TLC和HPLC）。结构 生物活性化合物的阐明将通过以下组合来实现 光谱（IR，UV，MS和NMR）技术，化学操作，或 X射线晶体学（如适用）。大规模生产（10-20毫克）的最多 将进行至3种选定的化合物以获得用于体内研究的材料。 在携带MCF-7的补充激素的SCID小鼠中的抗癌活性测试， T47 D人肿瘤异种移植物。 我们希望能够获得这种独特的生物活性天然来源， 产品，结合使用常规和功能性体外 生物测定，然后是体内测试将导致有效的发现， 可开发成临床有效药物的新抗癌剂 具有抗实体瘤活性。