DiversityExpanding the Synthetic Utility of a Flavin-dependent Monooxygenases

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 约80%的世界人口依赖于由天然产品制成的传统药物(REF)。这个 制药业有大约50%的药品是基于天然产品或从天然产品中提取的。不幸的是， 这些分子的合成通常非常复杂，需要安装多个官能团 具有非常特殊的3D结构的群体，对他们的生物活动至关重要。 酚类化合物的氧化脱芳构化是合成络合物的一种强有力的转化 分子，因为它引入了立体化学并产生了为进一步反应做好准备的产物。例如, 这一反应是异嗜色酮II和黄体素A生物合成的关键步骤。 HIV表面糖蛋白gp120与T细胞表面CD4的相互作用 是艾滋病毒入侵机制的核心。这种蛋白质-蛋白质相互作用的抑制被认为是为了破坏 人类免疫缺陷病毒(HIV)进入细胞。只有有限数量的对映体选择性 已经报道了氧化脱芳构化的方法，限制了这种转化在 合成化学。为了获得高度的立体选择性，化学计量的手性金属 除了长时间的反应时间、低温温度和苛刻的条件外，还需要络合物。 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、温和的反应条件和环境友好的使用 试剂。这些优势创造了对新的生物催化剂的需求，这些生物催化剂扩大了 合成化学家可以利用的产生复杂性的反应。然而，经常存在的权衡 生物催化剂的底物范围和选择性之间的关系限制了酶在合成中的应用。 根据这一建议，我们探索了一组FAD-单加氧酶、TropB、AfoD、AzaH和SorbC，其中包括 互补底物的范围和高水平的位置和立体选择性跨越一系列结构 不同的底物。 这项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了一条有价值的手性中间体的有效途径。这 研究将集中在Faf-单加氧酶、TropB、AfoD、AzaH和SorbC。如果我将(1)确定 酶面板的结合(2)将该信息转置以通过以下方式扩大TropB的底物范围 利用蛋白质工程对残基进行修饰(3)进一步提高生物催化剂的反应活性 利用C8-FAD类似物。这些工具将为有价值的手性中间体提供一条有效的途径 生物活性分子的合成。

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.