A Biosynthetic Strategy to Manufacture Less Toxic Amphotericins

生产毒性较小的两性霉素的生物合成策略

基本信息

批准号：
10383158
负责人：
Jonathan Webb Bogart
金额：
$ 2.45万
依托单位：
NORTHWESTERN UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-04-01 至 2022-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10383158
关键词：
Acquired Immunodeficiency Syndrome Amphotericin Amphotericin B Anabolism Antifungal Agents Artemisinins Back Breathing Cells Chemistry Clinical Complex Consumption Coupling Development Directed Molecular Evolution Dose-Limiting Elderly Engineering Enzymes Evolution Excision Gene Cluster Goals Guanosine Diphosphate Sugars Homology Modeling Hydrolase Hydrolysis Immune system Industrialization Infection Knowledge Learning Life Malaria Malignant Neoplasms Medical Medicine Mentors Methodology Microbe Modification Molds Mycoses Natural Products Nature New Acceptors Nucleotides Pathway interactions Patients Persons Pharmaceutical Preparations Pharmacologic Substance Polyenes Process Production Protein Biosynthesis Reaction Resistance Resources Route Science Site-Directed Mutagenesis Structure Structure-Activity Relationship Temperature Thermodynamics Time Toxic effect Training Tuberculosis Variant Water Work analog base career development catalyst chemical synthesis cost effective design effective therapy enzyme pathway flexibility glycosyltransferase improved innovation metabolic engineering mutant mycosamine nucleotidyltransferase pressure prevent protein structure prototype scaffold skills small molecule success sugar undergraduate student

项目摘要

PROJECT ABSTRACT Amphotericin B is a powerful antifungal treatment however, widespread use is prevented by dose limiting toxicity. Nontoxic analogs have been discovered but their structural complexity precludes a practical chemical synthesis on industrial scale. Microbes are extraordinarily adept at producing amphotericin, as metric tons are fermented annually. Enzymes found within amphotericin’s natural biosynthetic pathway can serve as exceptionally specific and renewable biocatalysts that can overcome many common limitations encountered during its chemical synthesis. Reimagining aspects of amphotericin’s biosynthesis could therefore produce a robust and sustainable path to these nontoxic variants. This proposal aims to develop a deep understanding of several catalytic processes involved in amphotericin’s natural biosynthesis. Harnessing this knowledge, key enzymes and biosynthetic pathways will be evolved and engineered to construct a biosynthetic strategy to manufacture less toxic amphotericins. Cell-free protein synthesis provides a means to prototype these enzymes and pathways in a quantitative and high-throughput manner. Ultimately this work is expected to be an efficient platform to create new, best-in-class antifungal candidates, provide probes to interrogate structure-activity relationships dictating AmB’s potency and toxicity, and drive the continued development of enzymes as renewable, ecologically-safe and flexible biocatalysts for pharmaceutical sciences. The proposal provides ample opportunities for me to build on the knowledge from my undergraduate and graduate studies so I may continue to learn, innovate, and develop skills to be a well-rounded scientific leader. Based on the unique scientific expertise and mentoring track record of my Sponsor and Cosponsor, they are the ideal pair to provide constructive guidance in science and career development, and maximize my potential to drive this project towards success.

项目摘要两性霉素 B 是一种强大的抗真菌治疗药物，但由于剂量限制性毒性而无法广泛使用。已发现无毒类似物，但其结构复杂性阻碍了实际的化学合成在工业规模上，微生物非常擅长生产两性霉素，因为发酵量为公吨。每年，在两性霉素的天然生物合成途径中发现的酶可以作为异常特异性的酶。和可再生生物催化剂，可以克服其化学过程中遇到的许多常见限制因此，重新构想两性霉素生物合成的各个方面可以产生稳健且可持续的方法。该提案旨在深入了解几种催化作用。利用这些知识、关键酶和两性霉素的自然生物合成过程。生物合成途径将得到进化和设计，以构建生物合成策略，以减少生产有毒两性霉素的无细胞蛋白质合成提供了一种对这些酶和途径进行原型设计的方法。最终，这项工作有望成为一个有效的平台。新型、一流的抗真菌候选药物，提供了探究结构-活性关系的探针 AmB 的效力和毒性，推动酶作为可再生、生态安全的持续发展该提案为我提供了充足的机会来构建药物科学。基于本科生和研究生学习的知识，以便我可以继续学习、创新和发展基于独特的科学专业知识和指导记录，培养成为全面的科学领导者的技能。我的赞助商和共同赞助商，他们是在科学和职业方面提供建设性指导的理想搭档发展，并最大限度地发挥我的潜力，推动该项目取得成功。