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; 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.

项目摘要 天然产品是药物灵感的重要来源。世界卫生组织估计 世界人口的约80％依赖于由天然产品制成的传统药物（参考）。这 制药行业约有50％的基于天然产品或源自天然产品的药物。不幸的是， 这些分子的合成通常是非常复杂的，需要安装多个功能 具有非常具体的3D体系结构对生物学活动至关重要的组。 酚类化合物的氧化亲爱的人体化是合成复合物的强大变化 分子引入立体化学并生成启动的产品以进行进一步反应。例如， 该反应是异粒元II和叶黄素A的生物合成的关键步骤，该抑制剂的抑制剂 GP120（在HIV表面上发现的糖蛋白和CD4）的相互作用在T细胞表面 HIV入侵机制的中心。假设这种蛋白质蛋白质相互作用的抑制以破坏 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 据报道了氧化性亲爱的方法的方法，限制了这种转化在 合成化学。为了达到高度的立体选择性，化学计量的手性金属 除了长时间的反应时间，低温温度和恶劣的条件外，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，轻度反应条件和环境良性的使用 试剂。这些优势已经对新的生物催化剂提出了需求，以扩大 合成化学家可用的复杂性产生反应。但是，经常存在的权衡 在生物催化剂的底物范围及其选择性之间限制了酶在合成中的应用。 在此提案中，我们探索了一组FAD-MONOOXYGASPase，Tropb，Afod，Azah和Sorbc，其中包含 互补的底物范围以及一系列结构上的高水平位点和立体选择性 不同的底物。 这项研究建议旨在开发具有互补选择性的一套催化剂，以便为了 为生物活性分子的合成提供有效的手性中间体的有效途径。这 研究将侧重于FAF单加氧酶，Tropb，Afod，Azah和Sorbc。如果我会（1）确定 酶面板的结合（2）转置此信息以扩大Tropb的底物范围 利用蛋白质工程的残基的修饰（3）进一步增强了生物催化剂的反应性 利用C8-FAD类似物。这样的工具将为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。