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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.

摘要糖肽类抗生素（Glycopeptide antibiotics，GPAs）是目前世界上最重要的抗生素之一。的该天然产物家族的创始成员万古霉素被用作治疗感染的最后药物，耐甲氧西林金黄色葡萄球菌（MRSA）和艰难梭菌。沿着其他一些抗生素，万古霉素提供了一个重要的武器，对“超级细菌”，致病菌，对多种临床抗生素获得性耐药。但是，随着对这最后一道防线的抵制蔓延，开发化学定制万古霉素和其他GPA的方法来创造新的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，但这一方法不适用于抗生素。这种方法在GPA中受到严格限制，主要是由于它们的化学复杂性和尺寸。的关键万古霉素的结构复杂性和生物活性是三个芳香交联，由两个芳基醚连接和联芳基碳-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。剩下的两个交叉点的来源，然而，仍然难以捉摸。我们最近发现，第二种P450酶OxyA引入了第二种P450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的酶活性，表明它安装了最终的联芳基连接，这是在任何GPA中首次演示此反应。此外，委员会认为，我们已经利用天然生物合成金属酶的反应性来实现化学酶促反应，产生万古霉素糖苷配基衍生物的途径。这个阶段是为了充分利用这种化学酶方法化学衍生万古霉素，希望产生有用的第二代衍生物。在本申请中，我们提出不仅完成万古霉素的化学-酶促合成，而且还包括已知保留生物活性的衍生物，即使是针对抗性病原体。我们进一步建议，建立万古霉素类似物的文库，我们称之为“设计师万古霉素”，含有修饰，是无法用现有的方法来实现的。我们将同时探索详细的化学机制的OxyB，并创建一个创新的固相方法，以提高我们的化疗的效率和可扩展性，酶促途径我们的研究将揭示万古霉素的生物合成，并使最全面的努力，但创造GPA变体具有独特的结构和可能的新的生物活性，通过优雅的化学-酶促途径