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

• 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
• 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

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是一种有效的抗真菌治疗药物，但由于剂量限制性毒性而无法广泛使用。 已经发现了无毒的类似物，但是它们的结构复杂性排除了实际的化学合成 工业规模。微生物非常善于生产抗生素，因为公吨发酵 每年。在阿氏菌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 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 合成.因此，重新构想两性霉素的生物合成方面可以产生强大且可持续的 找到这些无毒变异体的途径该建议旨在深入了解几种催化剂 这是一个涉及到植物素天然生物合成的过程。利用这些知识，关键酶和 生物合成途径将被进化和改造，以构建一种生物合成策略， 有毒的抗生素无细胞蛋白质合成提供了一种手段，原型这些酶和途径， 定量和高通量的方式。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，提供探针来询问结构-活性关系，决定 AmB的效力和毒性，并推动酶作为可再生的，生态安全的酶的持续发展 和制药科学的柔性生物催化剂。该提案为我提供了充分的机会， 在本科和研究生学习的基础上，我可以继续学习，创新和发展 成为一名全面的科学领导者的技能。基于独特的科学专业知识和指导记录， 作为我的赞助人和共同赞助人，他们是在科学和职业方面提供建设性指导的理想伴侣。 发展，并最大限度地发挥我的潜力，推动这个项目走向成功。