A Biosynthetic Strategy to Manufacture Less Toxic Amphotericins

项目摘要 两性霉素B是一种有效的抗真菌药物，但其广泛应用受到剂量限制毒性的阻碍。 已经发现了无毒的类似物，但它们的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，因为要发酵成吨。 每年一次。在两性霉素的自然生物合成途径中发现的酶可以起到异常特异的作用 和可再生生物催化剂，可以克服在其化学过程中遇到的许多常见限制 综合。因此，重新设想两性霉素的生物合成方面可以产生一个强大和可持续的 通向这些无毒变种的路径。这项建议旨在加深对几种催化作用的理解 两性霉素的自然生物合成过程。利用这一知识，关键的酶和 生物合成途径将被进化和设计，以构建一种生物合成策略，以减少生产 有毒的两性霉素。无细胞蛋白质合成提供了一种方法来制作这些酶和途径的原型 一种量化和高通量的方式。最终，这项工作有望成为一个高效的平台来创建 新的，同类中最好的抗真菌候选药物，提供了探查结构-活性关系的指示 AMB的效力和毒性，并推动酶的持续发展作为可再生的，生态安全的 以及用于制药科学的灵活的生物催化剂。这项提议为我提供了大量的机会来建立 在我的本科生和研究生学习的基础上，我可以继续学习，创新和发展 成为一名全面发展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 在我的赞助人和共同赞助人中，他们是在科学和职业生涯中提供建设性指导的理想组合 发展，并最大限度地发挥我的潜力，推动这个项目走向成功。

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.