High-throughput chemoenzymatic synthesis of bioactive compounds

提案摘要 随着耐药性感染和癌症变得越来越突出，研究新的耐药性药物是至关重要的。 来对抗这些日益严重的健康危机。氮杂菲酮，一类未充分研究的真菌天然 产品，已被证明表现出不同的生物学特性，使它们成为一类新的候选人， 治疗学虽然初步研究表明它们在这方面的效用，但对它们的进一步研究表明， 由于构建其密集功能化核心的挑战， 拥挤的立体中心此外，选择性地进入这些支架的C7构型， 探索阿扎菲隆结构-活性关系的额外障碍。 对这类分子的最简洁的方法依赖于氧化脱芳构化。 预官能化芳烃。不对称氧化脱芳构化中的现有技术方法需要 超化学计量的氧化剂和昂贵的手性配体。这些方法也只有 在有限的底物范围内得到证明，可能表现出较差的位点选择性，并需要强制反应 条件幸运的是，自然界已经进化出上级催化剂，以更大的催化活性进行氧化脱芳构化。 位点和立体选择性优于这些已建立的化学方法。很有希望的是，我们已经确定了两种黄素- 依赖性单加氧酶同系物，AzaH和AfoD，它们可以对 芳族底物具有相同的位点选择性，但提供了相反的立体化学构型， 随后生成阿扎菲隆产物。这一关键发现使它们能够用于合成各种天然药物， 产品和合成结构单元，提供正交位点和立体选择性， 更大的化学空间以环境友好的方式。此外，这些催化剂可以温和地操作， 反应条件，使其与高通量化合物生成平台相容。这 一项提案描述了在立体分散、化学酶促合成中使用这些生物催化剂的策略 用于快速生成阿扎菲隆类似物。 总之，这项工作的目的是通过高通量， 这些支架的战略多样化，由荧光偏振测定和细胞绘画指导 屏幕本文建立的方法将提供一种有效开发越来越有效的药物的手段。 基于阿扎菲隆的药物。

Proposal Summary With drug-resistant infections and cancers becoming more prominent, it is crucial to investigate new classes of small molecules to combat these growing health crises. Azaphilones, an underexplored class of fungal natural products, have been shown to exhibit diverse biological properties, making them candidates as a novel class of therapeutics. Although preliminary studies demonstrate their utility in this regard, further investigation of their activities has been hindered due to the challenges of constructing their densely functionalized core and congested stereocenter. Furthermore, selectively accessing either C7 configuration of these scaffolds presents an addition hurdle to exploring azaphilone structure-activity relationships. The most concise approach toward this class of molecules relies on the oxidative dearomatization of prefunctionalized arenes. Current state-of-the-art methods in asymmetric oxidative dearomatization require superstoichiometric quantities of both an oxidant and expensive chiral ligand. These methods have also only been demonstrated on a limited substrate scope, can exhibit poor site-selectivity, and require forcing reaction conditions. Fortunately, Nature has evolved superior catalysts to perform oxidative dearomatization with greater site- and stereoselectivity than these established chemical methods. Promisingly, we have identified two flavin- dependent monooxygenase homologs, AzaH and AfoD, which can perform an oxidative dearomatization on aromatic substrates with the same site-selectivity, but provide the opposite stereochemical configuration in the subsequent azaphilone product. This pivotal discovery enables their use in the synthesis of various natural products and synthetic building blocks, providing orthogonal site- and stereoselectivity to more readily access greater chemical space in an environmentally-benign manner. Furthermore, these catalysts can operate mild reaction conditions, making them compatible with high-throughput compound generation platforms. This proposal describes strategies for the use of these biocatalysts in stereodivergent, chemoenzymatic syntheses for the rapid generation of azaphilone analogs. In summary, this work aims to investigate azaphilone structure-activity relationships through high-throughput and strategic diversification of these scaffolds, directed by a fluorescence polarization assay and a cell painting screen. The methods established herein will provide a means to efficiently develop increasingly potent azaphilone-based drugs.