In Situ Natural Product Labeling and Applications

 说明(申请人提供)：天然产物及其衍生物一直是药物发现的丰富来源；据估计，市场上60%以上的药物来自天然来源。然而，在过去的三十年里，旨在利用天然产物进行药物发现的研究严重下降，部分原因是我们在新的天然产品发现和类似物生产方面的能力有限。迫切需要天然产物提纯、量化、过量生产和多样化的创新技术，以改变天然产物研究领域的范式，促进基于天然产物的药物发现和开发。由于标记天然产品可以解决天然产品研究的这些具有挑战性的方面，本提案试图开发一种新的策略，以克服已知方法的局限性，以标记具有生物正交功能的天然产品。为此，我们将探索不寻常的合成生物学，即 这些系统提供了生物正交功能，并开发了用于原位标记天然产品的通用平台，特别是难以合成或化学衍生的复杂天然产品。这一策略的成功将直接产生正交化的天然产物类似物，这些类似物可以进行简单的化学修饰来进行药物筛选。更重要的是，用独特的化学句柄标记天然产品将使我们能够通过生物正交化学对天然产品进行可视化、浓缩、量化和作用模式研究，从而对我们解决天然产品生物合成、生物学和药理学中具有挑战性的问题的能力产生深远的影响。此外，本文还提出了一种新的基于天然产物的、定量的、高通量的筛选方法，作为进化天然产物生物合成酶的有用工具，这些天然产物生物合成酶可以在特定的途径中高效地产生高滴度的天然产物类似物。这种筛选方法将被用来理解和设计几个重要的天然产物家族的生物合成，以用于新的抗生素和抗癌药物的发现。拟议的跨学科项目连接了天然产品生物合成、蛋白质工程和生物正交化学的多种创新，如果成功，将极大地扩展天然产品研究工具包，并在酶学、生物合成和医学方面产生广泛影响。因此，就其重要性、新颖性以及研究人员的创新和创造力记录而言，它是唯一适合新创新者奖的。该项目与研究组的其他工作几乎没有重叠，同时与各种研究项目提供了良好的协同作用，并完美地符合我们的主要研究主题之一：利用酶的力量进行天然产品的生物合成，最终应用于促进人类健康。

 DESCRIPTION (provided by applicant): Natural products and their derivatives have been rich sources for drug discovery; it has been estimated that more than 60% of the drugs that are in the market derive from natural sources. However, during the last three decades, research aimed at exploiting natural products for drug discovery has seriously declined, in part due to our limite capability in new natural product discovery and analogue production. Innovative technology for natural product purification, quantification, overproduction, and diversification is urgently neede to make a paradigm shift in the field of natural product research and to promote natural product-based drug discovery and development. As tagging natural products can address these challenging aspects of natural product research, this proposal seeks to develop a novel strategy that overcomes the limitations of known methods for tagging natural products with a bioorthogonal functionality. To this end, we will explore the unusual synthetic biology that living systems offer for bioorthogonal functionalities and develop general platforms for the in situ labeling of natural products, particularly complex natural products that are difficult to be synthesized or derivatized chemically. The success of this strategy will directly produce orthogonally functionalized natural product analogues that can be subjected to facile chemical modification for drug screening. More importantly, tagging natural products with a unique chemical handle will enable the visualization, enrichment, quantification, and mode of action study of natural products through bioorthogonal chemistry, and thus have a profound impact on our ability to address challenging questions in natural product biosynthesis, biology, and pharmacology. In addition, a novel natural product-based, quantitative, high-throughput screening method based on this tagging strategy is proposed here as a useful tool for evolving natural product biosynthetic enzymes that can efficiently function in specific pathways to produce natural product analogues in high titers. This screening method will be leveraged to understand and engineer the biosynthesis of several important families of natural products for new antibiotic and anticancer drug discovery. The proposed interdisciplinary project bridges multiple innovations in natural product biosynthesis, protein engineering, and bioorthogonal chemistry, and if successful, will vastly expand the natural product research toolkit and have broad impact in enzymology, biosynthesis, and medicine. It is therefore uniquely suited to the New Innovator Award with respect to its significance, novelty, and the investigator's record of innovativeness and creativity. This project has minimal overlap with other efforts in the research group, while providing good synergy with various research projects and fitting perfectly in one of our main research themes: to harness the power of enzymes towards the biosynthesis of natural products with ultimate applications in advancing human health.