Anticancer Agents from Plant- and Lichen-Associated Fungi of the Sonoran Desert

描述（由申请人提供）：天然产品对人类健康做出了巨大贡献。从天然来源中寻找新的先导化合物仍然是现代抗癌药物发现的关键因素。我们的长期目标是从索诺兰沙漠的植物和地衣相关真菌中发现通过新机制起作用的抗癌药物，从而进一步支持我们的假设，即在这种独特环境中相互关联的生物体精心设计了一种相对未被探索的强效生物活性化合物，以调节它们的相互作用并提高它们的存活率。这一跨学科和机构间项目的具体目标是：1。扩大索诺兰沙漠植物和地衣相关真菌的独特生物资源，优化500种选定真菌菌株的生物合成潜力，制备2500种提取物进行评估；2. 使用基于细胞的生物学和分子方法筛选抗癌活性提取物，包括：(a)在我们之前的资助期间建立的一种检测方法，最近进行了修改，以检测能够热休克诱导和抑制热休克诱导的化合物，随后进行了与Hsp90直接相互作用的检测，包括荧光素酶重折叠和使用热休克诱导实验分离的化合物的结合检测；(c)对亚细胞毒性浓度下的细胞运动抑制剂进行试验，(d)对五种前哨癌细胞系进行常规染料还原试验，以抑制增殖/存活，以避免发现一般的细胞毒性药物；3. 重复活性提取物，每年在多达五种有希望的真菌菌株上大规模培养，并将其提取物进行生物测定指导分离，以分离和表征生物活性化合物；和4。根据活性、结构新颖性和效力，利用基因组学和蛋白质组学方法确定最有前途的化合物的分子作用机制。我们期望这些最有趣的化合物将成为进一步优化开发具有独特作用机制的新型抗癌药物的先导。然而，在更基础的科学水平上，我们提出的创新化学生物学方法应该有助于确定(a)伴侣蛋白如Hsp90促进肿瘤发生的机制，以及(b)小分子抑制参与侵袭和转移的癌细胞迁移的机制，从而提高我们预防和治疗癌症的整体能力，从而对公众健康产生重大影响。

DESCRIPTION (provided by applicant): Natural products have made enormous contributions to human health. The search for new lead compounds from natural sources continues to be a crucial element in modern anticancer drug discovery. Our long-term goal is to discover anticancer drugs that act by novel mechanisms from plant- and lichen-associated fungi of the Sonoran desert, thereby providing further support for our hypothesis that organisms living in association with one another in this unique environment elaborate a relatively unexplored repertoire of potent bioactive compounds with which to mediate their interactions and enhance their survival. The specific aims of this interdisciplinary and inter-institutional project are to: 1. expand our unique bio-resource of Sonoran desert plant- and lichen-associated fungi and optimize biosynthetic potential of 500 selected fungal strains and prepare 2,500 extracts for evaluation; 2. screen extracts for anticancer activity using cell-based biological and molecular approaches including: (a) an assay established during our previous grant period and recently modified to detect compounds capable of heat shock induction and inhibition of heat shock induction, followed by assays for direct interaction with Hsp90 including luciferase-refolding and binding assays for compounds isolated using the heat shock induction assay; (c) an assay for inhibitors of cell motility at sub-cytotoxic concentrations, and (d) a conventional dye reduction assay for inhibition of proliferation/survival based on five sentinel cancer cell lines to avoid the discovery of general cytotoxic agents; 3. dereplicate active extracts, culture on large-scale up to five promising fungal strains per year and subject their extracts to bioassay-guided fractionation to isolate and characterize bioactive compounds; and 4. define molecular mechanisms of action using genomic and proteomic approaches for the most promising compounds based on their activity, structural novelty, and potency. We expect that the most interesting compounds will serve as leads for further optimization in the development of novel anticancer agents with unique mechanisms of action. At a more basic scientific level, however, the innovative chemical biology approach we propose should help define the mechanisms by which (a) chaperones such as Hsp90 facilitate oncogenesis, and (b) small-molecules inhibit cancer cell migration involved in invasion and metastasis, leading to enhancement of our overall ability to prevent and cure cancers, and therefore have a great impact on public health.