Bioactivity Guided Marine Natural Product Discovery Using Phenotypic Assays

使用表型测定的生物活性引导海洋天然产品发现

• 批准号: RGPIN-2018-03817
• 负责人: Andersen, Raymond
• 金额: $ 3.5万
• 依托单位: University of British Columbia
• 依托单位国家: 加拿大
• 项目类别: Discovery Grants Program - Individual
• 财政年份: 2020
• 资助国家: 加拿大
• 起止时间: 2020-01-01 至 2021-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=711866
• 关键词: Bioactivity; Guided; Marine; Natural; Product

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.

拟议研究的主要目标将是使用Forward Chemical Genetics基于细胞的表型分析筛选平台，以促进发现结构新颖的生物活性海洋天然产物（MNP），这些产物可用作基础生物学研究的工具，包括i）MHC-1抗原表达，ii）血管完整性和iii）基因MYC的表达。第二个目标将是选择的MNP和类似物的化学合成，以定义其药效团的结构活性关系（SAR），产生用于蛋白质靶标鉴定的探针，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合作。我们的合作者开发了用于MNP发现计划的表型测定，以找到他们研究健康人类生物学中重要的基本过程所需的化学工具。天然产物的发现和合成是基础有机化学NSE研究的核心活动。本提案中要求的资金仅用于支持Andersen集团的天然产品化学活动。发现生物测定，蛋白质靶点鉴定和其他生物学研究将在我们合作者的实验室进行，由他们的独立赠款。 新的MNP的发现既不是常规的，也不是微不足道的。经验告诉我们，使用基于细胞的生物筛选寻找新的非细胞毒性表型反应的正向化学遗传学方法是发现真正独特的MNP的最有效方法之一，这些MNP对有机化学界具有重大意义，并具有作为细胞生物学工具的实用价值。化学遗传学使用小分子来改变细胞环境中单个基因产物的功能，以便将特定基因和基因产物与表型联系起来。正向化学遗传学筛选在基于细胞或整个生物体测定中引发感兴趣表型的分子，然后鉴定蛋白质靶标。我们的海洋无脊椎动物和培养微生物的粗提物库将是新的小分子蛋白质配体的来源。研究的意义是多方面的。新MNP的发现为已知化学实体的库增加了独特的多样性，并揭示了未知的生物合成途径和药效团。复杂的生物活性MNP是合成有机化学家的有吸引力的目标，推动了新的合成策略和方法的发展。发现MNP调节涉及MHC-1表达、血管完整性和MYC表达的蛋白质，将为研究单细胞和动物中这些重要的生物学过程提供工具，并对其正常调控网络产生更深入的了解。我们的跨学科研究将提供培训，使HQP能够获得高层次的工业和学术工作。