Bioactivity Guided Marine Natural Product Discovery Using Phenotypic Assays

拟议研究的主要目的是使用基于细胞的正向化学遗传学表型分析筛选平台，以促进发现结构新颖的生物活性海洋天然产物（MNPs），这些产物可作为基础生物学研究的工具，用于i) MHC-1抗原表达，ii)血管完整性和iii) MYC基因表达。次要目标将是选定的MNPs和类似物的化学合成，以确定其药效团的结构活性关系（SAR），生成用于蛋白质靶点鉴定的探针，并为生物学研究提供材料和类似物。拟议的研究是跨学科和协作的。我们的合作者开发了用于MNP发现计划的表型分析，以寻找他们需要的化学工具来研究健康人类生物学中重要的基本过程。天然产物的发现与合成是基础有机化学NSE研究的核心活动。本提案所要求的资金将仅用于支持安达信集团的天然产物化学活动。发现生物分析，蛋白质靶标鉴定和其他生物学研究将在我们的合作伙伴的实验室进行，这些实验室由他们的独立研究经费资助。

The main objective of the proposed research will be to use Forward Chemical Genetics cell-based phenotypic assay screening platforms to facilitate the discovery of structurally novel bioactive marine natural products (MNPs) that can be used as tools in basic biological studies of i) MHC-1 antigen expression, ii) vascular integrity, and iii) expression of the gene MYC. A secondary objective will be the chemical synthesis of selected MNPs and analogs to define the Structure Activity Relationship (SAR) for their pharmacophores, generate probes for protein target identification, and provide material and analogs for biological studies. The proposed research is interdisciplinary and collaborative. Phenotypic assays to be used in the MNP discovery program have been developed by our collaborators to find the chemical tools they require to study fundamental processes that are important in healthy human biology. Natural products discovery and synthesis are core activities of basic organic chemistry NSE research. Funds requested in this proposal will only support the natural products chemistry activities in Andersen's group. The discovery bioassays, protein target identification, and other biological studies will take place in our collaborator's labs funded by their independent research grants. The discovery of novel MNPs is neither routine or trivial. Experience has taught us that a Forward Chemical Genetics approach using cell-based biological screens looking for novel non-cytotoxic phenotypic responses is one of the most effective ways to discover truly unique MNPs that are of significant interest to the organic chemistry community and have practical value as cell biology tools. Chemical genetics uses small molecules to alter the function of a single gene product within a cellular context in order to link a specific gene and gene product to a phenotype. Forward chemical genetics screens for molecules that elicit a phenotype of interest in a cell-based or whole organism assay and then identifies the protein target. Our library of crude extracts of marine invertebrates and cultured microorganisms will be the source of novel small molecule protein ligands. The significance of the research will be multifold. Discovery of new MNPs adds unique diversity to the pool of known chemical entities and reveals unknown biosynthetic pathways and pharmacophores. Complex bioactive MNPs are attractive targets for synthetic organic chemists, driving the development of new synthetic strategies and methodology. Discovery of MNPs that modulate proteins involved MHC-1 expression, vascular integrity, and expression of MYC will provide tools to study these important biological processes in single cells and animals and generate deeper understanding of their normal regulatory networks. Our interdisciplinary research will provide training that will enable HQP to obtain high-level industrial and academic jobs.