Toward a Chemo-Enzymatic Synthesis of Vancomycin and Its Analogs

万古霉素及其类似物的化学酶法合成

• 批准号: 10170408
• 负责人: Mohammad R Seyedsayamdost
• 金额: $ 31.03万
• 依托单位: PRINCETON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-07-01 至 2023-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10170408
• 关键词: Address; Amino Acids; Anabolism; Antibiotic Resistance; Antibiotics; Biochemical Reaction; Biogenesis; Biological; Carbon; Chemicals; Chemistry; Clinical; Clostridium difficile; Complex; Cytochrome P450; Cytochrome a; Cytochromes; Data; Engineering; Enzymes; Ethers; Family; Generations; Glycopeptide Antibiotics; Glycopeptides; Goals; Heme; In Vitro; Infection; Knowledge; Libraries; Ligation; Light; Methodology; Methods; Modification; Natural Products; Nature; Oxides; Peptide Synthesis; Peptides; Peroxidases; Pharmaceutical Preparations; Phase; Phenols; Plant Resins; Production; Property; Reaction; Reporting; Research; Resistance; Resort; Role; Route; Scheme; Scientist; Series; Solid; Solvents; Structure; Superbug; Surface; Therapeutic Agents; Time; Vancomycin; Variant; analog; base; catalyst; cofactor; crosslink; experimental study; fascinate; global health; glycosylation; improved; innovation; member; metalloenzyme; methicillin resistant Staphylococcus aureus; novel; pathogen; pathogenic bacteria; resistance mechanism; stem; weapons

ABSTRACT Glycopeptide antibiotics (GPAs) are among the most important therapeutic agents world-wide. The founding member of this natural product family, vancomycin, is used a drug of last resort against infections by methicillin-resistant Staphylococcus aureus (MRSA) and Clostridium difficile. Along with a handful of other antibiotics, vancomycin provides an important weapon against “superbugs”, pathogenic bacteria that have acquired resistance to multiple clinical antibiotics. But as resistance to even this last line of defense spreads, it is ever more important to develop means of chemically tailoring vancomycin and other GPAs to create new derivatives that counter known resistance mechanisms. Synthetic derivatization has proven to be a successful method for creating new antibiotics, but this approach is severely restricted within the GPAs, primarily due to their chemical complexity and size. Key to the structural complexity and biological activity of vancomycin are three aromatic crosslinks, consisting of two aryl ether connections and a biaryl carbon-carbon bond. Research over the past 20 years has shown that a cytochrome P450 enzyme (OxyB) installs the first aryl ether bond. The origin of the remaining two crosslinks, however, remained elusive. We recently showed that OxyA, a second P450 enzyme, introduces the second aryl ether crosslink during vancomycin biogenesis. We further recapitulated the enzymatic activity of OxyC and showed that it installs the final biaryl connection, the first demonstration of this reaction in any GPA. Moreover, we have exploited the reactivities of the native biosynthetic metalloenzymes to implement a chemo-enzymatic route for creating a vancomycin aglycone derivative. The stage is set to fully leverage this chemo-enzymatic approach to chemically derivatize vancomycin in the hopes of generating useful second-generation derivatives. In the current application, we propose to complete the chemo-enzymatic synthesis of not just vancomycin, but also of derivatives known to retain bioactivity, even against resistant pathogens. We further propose to build a library of vancomycin analogs that we refer to as “designer vancomycins”, containing modifications that are inaccessible with current methodologies. We will simultaneously explore the detailed chemical mechanism of OxyB and create an innovative solid-phase approach to enhance the efficiency and scalability of our chemo- enzymatic route. Our studies will shed light onto the biosynthesis of vancomycin and enable the most comprehensive effort yet to create GPA variants with unique structures and possibly new bioactivities via an elegant chemo-enzymatic route.

抽象的 糖肽抗生素（GPA）是世界范围内最重要的治疗药物之一。 万古霉素是这个天然产品家族的创始成员，被用作对抗感染的最后手段 耐甲氧西林金黄色葡萄球菌 (MRSA) 和艰难梭菌以及其他一些细菌。 作为抗生素，万古霉素提供了对抗“超级细菌”的重要武器，“超级细菌”是一种致病细菌。 但随着对这最后一道防线的耐药性的蔓延，它也对多种临床抗生素产生了耐药性。 开发化学定制万古霉素和其他 GPA 的方法来创造新的药物变得越来越重要 对抗已知耐药机制的衍生物。 合成衍生化已被证明是创造新抗生素的成功方法，但这 方法在 GPA 中受到严格限制，主要是由于其化学复杂性和尺寸。 万古霉素的结构复杂性和生物活性是三个芳香族交联键，由两个芳基组成 过去20年的研究表明，醚连接和联芳基碳-碳键。 细胞色素 P450 酶 (OxyB) 安装第一个芳基醚键 其余两个交联的起源， 然而，我们最近发现第二种 P450 酶 OxyA 引入了第二种酶。 我们进一步概括了 OxyC 和万古霉素生物合成过程中芳基醚交联的酶活性。 表明它安装了最终的联芳基连接，这是任何 GPA 中该反应的首次演示。 我们利用天然生物合成金属酶的反应性来实施化学酶促 创造万古霉素苷元衍生物的路线已准备好充分利用这种化学酶。 化学衍生万古霉素的方法，希望产生有用的第二代衍生物。 在当前的应用中，我们建议完成的不仅仅是万古霉素的化学酶合成， 而且还具有已知保留生物活性的衍生物，甚至针对耐药病原体。 建立一个万古霉素类似物库，我们称之为“设计万古霉素”，其中包含以下修饰： 目前的方法无法实现，我们将同时探索详细的化学机制。 OxyB 并创建一种创新的固相方法来提高我们化学的效率和可扩展性 我们的研究将揭示万古霉素的生物合成并实现最大程度的利用。 尚未通过全面的努力来创造具有独特结构和可能的新生物活性的 GPA 变体 优雅的化学酶途径。