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

基本信息

批准号：
10439760
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
项目类别：
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/10439760
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

项目摘要

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）是全球最重要的治疗剂之一。这该天然产品家族的创始成员万古霉素被用来抗衡感染的药物耐甲氧西林金黄色葡萄球菌（MRSA）和艰难梭菌。以及少数其他抗生素，万古霉素为“超级细菌”，致病细菌提供了重要的武器获得了多种临床抗生素的耐药性。但是，随着对最后一条防线的抵抗，它也会传播开发化学定制万古霉素和其他GPA的方法越重要，以创建新的抵抗已知抗性机制的衍生物。事实证明，合成衍生化是创建新抗生素的成功方法，但这方法在GPA中受到严格限制，这主要是由于它们的化学复杂性和大小。关键万古霉素的结构复杂性和生物学活性是三个芳族交联，由两个芳基组成醚连接和双碳碳键。过去20年的研究表明细胞色素P450酶（OxyB）安装第一个芳基醚键。其余两个交联的起源，但是，仍然难以捉摸。我们最近表明，第二个P450酶OxyA引入了第二个酶万古霉素生物发生过程中的芳基醚交联。我们进一步概括了Oxyc和Oxyc的酶活性表明它安装了最终的双轴连接，这是任何GPA中此反应的第一个演示。而且，我们探索了天然生物合成金属酶的反应性以实施化学酶创建万古霉素aglamcone衍生物的途径。该阶段将充分利用这种化学酶化学衍生化万古霉素的方法，希望产生有用的第二代衍生物。在当前的应用中，我们建议完成万古霉素的化学酶合成但也有衍生物已知可以保留生物活性，甚至抵抗抗性病原体。我们进一步建议建立一个我们称为“设计师万古霉素”的万古霉素类似物的库，其中包含修改与当前方法无法访问。我们将简单地探索详细的化学机制氧气并创建创新的固相方法，以提高我们的化学效率和可伸缩性酶促路线。我们的研究将阐明万古霉素的生物合成，并最大程度地启用全面的努力尚未创建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。