Selective P450 Oxidation Catalysts for Synthesis of Bioactive Molecules

用于合成生物活性分子的选择性 P450 氧化催化剂

• 批准号: 8643262
• 负责人: Rudi Fasan
• 金额: $ 27.04万
• 依托单位: UNIVERSITY OF ROCHESTER
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-06-01 至 2017-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8643262
• 关键词: Active Sites; Adopted; Antimalarials; Artemisinins; Binding; Biological Assay; Biological Factors; Catalysis; Cells; Chemistry; Complex; Coupling; Cytochrome P450; Development; Engineering; Enzymes; Evaluation; Fingerprint; Generations; Geometry; Glean; Goals; Human; Hydrogen Bonding; Hydroxylation; Libraries; Malaria; Maps; Mediating; Membrane; Metabolic; Methodology; Methods; Mixed Function Oxygenases; Mutagenesis; Nature; Organic Chemistry; Outcome; Parasites; Pharmaceutical Chemistry; Plasmodium falciparum; Positioning Attribute; Property; Protein Engineering; Protein Fingerprints; Research; Scheme; Shapes; Site; Staging; Structure; Structure-Activity Relationship; Supporting Cell; Terpenes; Testing; Time; Ursidae Family; Variant; anticancer activity; antileukemic agent; artemisinine; base; catalyst; chemical synthesis; clinically relevant; drug development; drug discovery; high throughput screening; improved; insight; leukemia; leukemic stem cell; novel; oxidation; parthenolide; scaffold; tool

DESCRIPTION (provided by applicant): The selective functionalization of aliphatic C-H bonds in complex organic molecules remains a most challenging problem in organic chemistry. Development of efficient methods to afford this transformation, in particular in the context of biologically active natural products, is bound to disclose unprecedented opportunities in the transformation of these molecules, thereby accelerating drug discovery efforts. In this application, we propose research aimed at developing powerful new methodologies to exploit P450 C-H oxidation catalysis and P450-mediated synthesis for selective transformation of unreactive aliphatic C-H bonds in complex natural products. To this end, efficient and time-effective strategies will be implemented to enable tailoring of the substrate reactivity, regio- an stereoselectivity of P450 monooxygenases and rapid generation of selective P450 catalysts for mediating functionalization of distinct unactivated aliphatic sites within a complex organic scaffold. At the basis of the proposed strategies there is a new enabling methodology to map the active site configuration in P450 monooxygenases and rapidly acquire information-rich functional profiles of these enzymes ('P450 fingerprints'). Systematic methods will be implemented to predict the reactivity of P450 variants via analysis of their fingerprints and to identify optimal mutagenesis schemes for altering the active site configuration in P450 monooxygenases without disrupting catalytic function. These new tools will be applied to obtain P450 catalysts with tailor-made regio- and stereoselectivity for hydroxylation of multiple aliphati positions in terpenes with clinically relevant anticancer and antimalarial activity. These efforts will unlock new avenues for late-stage transformation of these molecules in order to improve their pharmacological properties. By coupling selective P450-catalyzed aliphatic hydroxylations to chemical synthesis, novel and currently inaccessible derivatives of these natural products will be made available for activity evaluation. Through these studies, comprehensive structure-activity insights will be gleaned on these natural products, providing a basis for development of more potent antileukemic and antimalarial agents. Completion of the primary objectives of this application will result in a set of powerful and general strategies for P450 C-H oxidation catalyst development which will be readily applicable to a variety of other natural products and high- value compounds.

描述（由申请人提供）：复杂有机分子中脂肪族C-H键的选择性官能化仍然是有机化学中最具挑战性的问题。开发有效的方法来实现这种转化，特别是在生物活性天然产物的背景下，必将在这些分子的转化中揭示前所未有的机会，从而加速药物发现工作。在此应用中，我们提出的研究旨在开发强大的新方法，以利用 P450 C-H 氧化催化和 P450 介导的合成来选择性转化复杂天然产物中的非反应性脂肪族 C-H 键。为此，将实施高效且省时的策略，以实现底物反应性、P450单加氧酶的区域选择性和快速生成选择性P450催化剂的定制，以介导复杂有机支架内不同未活化脂肪族位点的功能化。在所提出的策略的基础上，有一种新的方法可以绘制 P450 单加氧酶的活性位点配置图，并快速获取这些酶的信息丰富的功能图谱（“P450 指纹”）。将实施系统方法，通过分析指纹来预测 P450 变体的反应性，并确定最佳诱变方案，以在不破坏催化功能的情况下改变 P450 单加氧酶的活性位点配置。这些新工具将用于获得具有定制区域和立体选择性的 P450 催化剂，用于萜烯中多个脂肪族位置的羟基化，具有临床相关的抗癌和抗疟活性。这些努力将为这些分子的后期转化开辟新途径，以改善其药理学特性。通过将选择性 P450 催化的脂肪族羟基化与化学合成相结合，这些天然产物的新型且目前无法获得的衍生物将可用于活性评估。通过这些研究，将收集对这些天然产物的全面结构活性见解，为开发更有效的抗白血病和抗疟药物奠定基础。完成本应用的主要目标将产生一套针对 P450 C-H 氧化催化剂的强大且通用的策略 开发将很容易适用于各种其他天然产品和高价值化合物。