isolation, structure elucidation and synthesis of biologically active marine natural products

该研究的目的是发现结构新颖的海洋天然产物，可用作细胞生物学和自噬，miRNA基因沉默和Toll样受体-5（TLR 5）信号转导的整体动物研究中的化学工具。有机合成将用于研究新药效团的SAR，制备用于蛋白质靶标鉴定的生物素化探针，并为动物研究提供充足的材料。采用基于细胞的测定来筛选化学文库以获得产生感兴趣的表型的基因产物（蛋白质）功能的无毒小分子调节剂的“正向化学遗传学”方法将用于促进新的海洋天然产物细胞生物学工具的发现。选择生物系统，自噬，miRNA基因沉默和toll样受体-5信号传导，是因为：i）它们最近被认识到在基础细胞生物学和人类疾病中发挥重要作用（神经变性和囊性纤维化），ii）所有这三种在遗传和蛋白质水平上的特征都不完全，和iii）对于其中两种没有已知的小分子调节剂，对于第三种只有非选择性调节剂。来自巴布亚新几内亚、印度尼西亚和多米尼加的热带栖息地和来自不列颠哥伦比亚的冷水栖息地的海洋无脊椎动物和微生物将作为海洋天然产物发现/化学遗传学研究的丰富和相对未开发的新化学多样性资源。该研究将提供新的分子工具，以研究单细胞和动物在遗传和蛋白质水平上的重要生物学过程，帮助确定新的药物靶点，并可能为开发治疗神经变性和囊性纤维化的药物提供先导化合物。新的生物活性天然产物的发现增加了独特的多样性，已知的化学实体的池，揭示未知的生物合成途径和药效团，并为合成有机化学家提供有吸引力的目标，刺激新的合成策略和方法的发展。

The objective of the proposed research will be to discover structurally novel marine natural products that can be used as chemical tools in cell biology and whole animal investigations of autophagy, miRNA gene silencing, and toll-like-receptor-5 (TLR5) signaling. Organic synthesis will be used to investigate the SAR of the new pharmacophores, to make biotinylated probes for protein target identification, and provide sufficient material for animal studies. A 'Forward Chemical Genetics' approach that employs cell-based assays to screen chemical libraries for non-toxic small molecule modulators of gene product (protein) function that produce phenotypes of interest will be used to facilitate the discovery of new marine natural product cell biology tools. The biological systems, autophagy, miRNA gene silencing, and toll-like-receptor-5 signaling, have been chosen because: i) they have recently been recognized to play important roles in basic cell biology and human disease (neurodegeneration and cystic fibrosis), ii) all three are incompletely characterized at the genetic and protein level, and iii) there are no small molecule modulators known for two of them and only non-selective modulators available for the third. Marine invertebrates and microorganisms from tropical habitats in Papua New Guinea, Indonesia, and Dominica and from cold water habitats in BC will serve as a rich and relatively unexplored resource of novel chemical diversity for this marine natural products discovery/chemical genetics research. The research will provide new molecular tools to study important biological processes at the genetic and protein level in single cells and animals, help identify new drug targets, and may provide lead compounds for development of drugs for the treatment of neurodegeneration and cystic fibrosis. Discovery of new bioactive natural products adds unique diversity to the pool of known chemical entities, reveals unknown biosynthetic pathways and pharmacophores, and provides attractive targets for synthetic organic chemists, stimulating the development of new synthetic strategies and methodology.