DiversityExpanding the Synthetic Utility of a Flavin-dependent Monooxygenases

多样性扩展黄素依赖性单加氧酶的合成效用

• 批准号: 10063422
• 负责人: Attabey Rodríguez Benítez
• 金额: $ 2.25万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-08-01 至 2021-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10063422
• 关键词: 3-Dimensional; Ache; Active Sites; Amino Acids; Anabolism; Architecture; Benign; Binding; Biochemical; Biological; Biomimetics; Breathing; CD4 Antigens; Catalysis; Cells; Chemicals; Complex; Crystallization; Data; Development; Drug Industry; Electrons; Engineering; Enzymes; Flavins; Flavoproteins; Glycoproteins; HIV; HIV Envelope Protein gp120; Human; Life; Lymphocyte; Mediating; Medicine; Membrane Glycoproteins; Methods; Mixed Function Oxygenases; Modification; Molecular; Molecular Probes; Natural Products; Nature; Oxidants; Oxygen; Peripheral; Pharmaceutical Preparations; Phenols; Population; Property; Protein Engineering; Protein Inhibition; Reaction; Reaction Time; Reagent; Reporting; Research; Research Proposals; Role; Route; Savings; Site; Source; Structure; Surface; Synthesis Chemistry; System; T-Lymphocyte; Temperature; Testing; Therapeutic; Traditional Medicine; Work; World Health Organization; analog; base; catalyst; cofactor; cryogenics; flavin-containing monooxygenase; functional group; improved; inhibitor/antagonist; insight; interest; metal complex; molecular dynamics; phenolate; protein protein interaction; stereochemistry; tool

Project Summary Natural products are an essential source of inspiration for medicines. The World Health Organization estimates that ~80% of the world's population relies on traditional medicines made from natural products (ref). The Pharmaceutical industry has ~50% of all drugs based on or derived from natural products. Unfortunately, the synthesis of these molecules is often prohibitively complex, requiring the installation of multiple functional groups with very specific 3D architecture critical to their biological activity. Oxidative dearomatization of phenolic compounds is a powerful transformation for the synthesis of complex molecules, as it introduces stereochemistry and generates products primed for further reaction. For example, this reaction is the key step in the biosynthesis of isochromophilone II and luteusin A, inhibitors of the interaction between gp120, a glycoprotein found on the surface of HIV, and CD4, on the surface of T-cells central to the HIV invasion mechanism. Inhibition of this protein-protein interaction is hypothesized to disrupt the entry of the human immunodeficiency virus (HIV) into cells. Only a limited number of enantioselective methods have been reported for oxidative dearomatization, limiting the application of this transformation in synthetic chemistry. To achieve high degrees of stereoselectivity, stoichiometric amounts of the chiral metal complexes are required in addition to lengthy reaction times, cryogenic temperatures, and harsh conditions. Biocatalytic reactions embody many features of ideal chemical transformations, including the potential for impeccable selectivity, high catalytic efficiency, mild reaction conditions and the use of environmentally benign reagents. These advantages have created a demand for new biocatalysts that expand the portfolio of complexity-generating reactions available to synthetic chemists. However, the tradeoff that often exists between the substrate scope of a biocatalyst and its selectivity limits the application of enzymes in synthesis. On this proposal, we explore a panel of FAD-monooxygenases, TropB, AfoD, AzaH, and SorbC containing complementary substrates scopes and high levels of site- and stereoselectivity across a range of structurally diverse substrates. This research proposal aims to aim to develop a suite of catalysts with complementary selectivities in order to provide an efficient route to valuable chiral intermediates for the synthesis of bioactive molecules. This research will focus on FAF-monooxygenases, TropB, AfoD, AzaH, and SorbC. Were I will (1) Determine the binding of the panel of enzymes (2) Transpose this information to expand the substrate scope of TropB by modification of residues utilizing protein engineering (3) Further enhance the reactivity of the biocatalyst by utilizing C8-FAD analogs. Such tools will provide an efficient route to valuable chiral intermediates for the synthesis of bioactive molecules.

项目概要 天然产品是药物灵感的重要来源。世界卫生组织估计 世界上约 80% 的人口依赖天然产品制成的传统药物（参考文献）。这 制药行业约 50% 的药物基于或衍生自天然产品。不幸的是， 这些分子的合成通常非常复杂，需要安装多功能 具有对其生物活性至关重要的非常特定的 3D 架构的群体。 酚类化合物的氧化脱芳构化是复杂化合物合成的有力转化 分子，因为它引入了立体化学并生成了用于进一步反应的产物。例如， 该反应是异色素 II 和叶黄体素 A 生物合成的关键步骤，它们是 gp120（HIV 表面的一种糖蛋白）与 T 细胞表面的 CD4 之间的相互作用 HIV入侵机制的核心。假设抑制这种蛋白质-蛋白质相互作用会破坏 人类免疫缺陷病毒（HIV）进入细胞。只有有限数量的对映选择性 已经报道了氧化脱芳构化的方法，限制了这种转化的应用 合成化学。为了实现高度的立体选择性，手性金属的化学计量量 除了漫长的反应时间、低温和恶劣的条件之外，还需要配合物。 生物催化反应体现了理想化学转化的许多特征，包括 无可挑剔的选择性、高催化效率、温和的反应条件以及使用对环境无害的 试剂。这些优势创造了对新型生物催化剂的需求，从而扩大了产品组合 合成化学家可以利用产生复杂性的反应。然而，经常存在的权衡 生物催化剂的底物范围及其选择性之间的限制限制了酶在合成中的应用。 在这个提案中，我们探索了一组 FAD-单加氧酶、TropB、AfoD、AzaH 和 SorbC，其中包含 互补底物范围以及跨一系列结构的高水平位点和立体选择性 多样化的基材。 该研究计划旨在开发一套具有互补选择性的催化剂，以便 为合成生物活性分子提供有价值的手性中间体的有效途径。这 研究将集中于 FAF-单加氧酶、TropB、AfoD、AzaH 和 SorbC。如果我将 (1) 确定 酶组的结合 (2) 转置此信息以扩展 TropB 的底物范围 利用蛋白质工程对残基进行修饰 (3) 进一步增强生物催化剂的反应活性 利用 C8-FAD 类似物。这些工具将为获得有价值的手性中间体提供有效的途径 生物活性分子的合成。