Studies to understand and exploit 10-membered enediyne biosynthesis

摘要 10-元烯二炔[例如加利车霉素（CLM）、埃斯帕霉素（ESP）和动力霉素（DYN）] 可以说是迄今为止发现的最著名的天然产品（NP）之一， 前所未有的复杂分子结构，显著的抗癌和抗感染效力， 的CLM，证明了临床实用性。目前的研究建立在长期合作的基础上， 与10元烯二炔生物合成的关键方面有关的成就和发现以及平行的 创新的努力，以增选关键的生物合成催化剂的合成应用。提出的研究将采取 这种强大的基础和遗传，生化，化学和蛋白质的强大组合的优势 结构工具来阐明剩余的不寻常的生物合成转化并利用选择的催化剂 烯二炔非天然修饰。具体而言，目标1和2将侧重于扩大我们对 烯二炔核心生物合成的基本步骤与CLM/DYN/ESP、DYN蒽醌生物合成相同 和一组选定的独特的剪裁反应（CLM/ESP硫代糖硫安装和氨基戊糖N- 烷基化、ESP C6-羟基化和0-糖基化）。与此同时，aim 3将侧重于战术结构研究 加强目标1和目标2以及“未知数”的结构研究，以便于功能注释。额外 目的2中的研究采用关键催化剂和相应的非天然底物， 用于化学选择性手柄的战略性安装，以实现用于以下的简单、温和的生物缀合的新方法： CLM针对肿瘤靶向mAb（与辉瑞合作）。

ABSTRACT The 10-membered enediynes [exemplified by calicheamicin (CLM), esperamicin (ESP) and dynemicins (DYN)] are arguably among the most renowned natural products (NPs) discovered to date by virtue of their unprecedented complex molecular architectures, notable anticancer and anti-infective potencies and, in the case of CLM, demonstrated clinical utility. The current study builds on a longstanding collaborative effort of achievement and discovery relating to key aspects of 10-membered enediyne biosynthesis as well as parallel innovative efforts to co-opt key biosynthetic catalysts for synthetic applications. The studies put forth will take advantage of this strong foundation and a powerful combination of genetic, biochemical, chemical and protein structural tools to elucidate the remaining unusual biosynthetic transformations and to exploit select catalysts for enediyne non-native modification. Specifically, aims 1 and 2 will focus on extending our understanding of the fundamental steps of enediyne core biosynthesis common to CLM/DYN/ESP, DYN anthraquinone biosynthesis and a selected set of unique tailoring reactions (CLM/ESP thiosugar sulfur installation and aminopentose N- alkylation, ESP C6-hydroxylation and O-glycosylation). In parallel, aim 3 will focus on tactical structural studies to augment both aims 1 and 2 and the structural study of ‘unknowns’ to facilitate functional annotation. Additional studies in aim 2 with key catalysts and corresponding non-native substrates are designed to assess the potential for strategic installation of chemoselective handles to enable novel approaches for facile, mild bioconjugation of CLM to tumor-targeting mAbs (in collaboration with Pfizer).