Abiological enzymatic C-H functionalization for bioactive molecule construction and diversification

项目摘要/摘要 抗生素是一种通过杀死细菌或抑制细菌来治疗细菌感染的抗菌药物。 生长以允许免疫清除。虽然抗生素有助于治疗感染，但它们的使用给患者带来了选择性压力 细菌并不可避免地导致耐药细菌的出现。此外，小说的发展 近年来，抗生素的使用速度有所放缓，这使得我们对多重耐药的治疗选择变少了 细菌感染。此外，这些有问题的多重耐药细菌大多是革兰氏阴性菌， 这意味着它们是双层的，因此对许多药物具有内在抗药性，使药物开发 更具挑战性。迫切需要创新的方法来开发新的抗生素，这些抗生素 对革兰氏阴性细菌有效，这样我们就可以保持我们治疗这些感染的能力。前景看好 研究表明，添加伯胺可以改善药物在革兰氏阴性菌中的积聚。 细菌为多种化合物，使其成为一种合理可行的提高活性的途径，前景看好 抗菌药物通过改善其细菌渗透性。合成胺化生物活性化合物可以是 繁琐，通常需要多个步骤和添加其他官能团才能获得所需的 阿米恩。相反，细胞色素P450的定向进化是开发酶的一种有前途的策略 生物活性化合物的后期修饰平台。P450酶的功能化能力 生物活性化合物中的惰性但普遍存在的C-H键可以被利用来促进 天然产品。最近的成功扩大了P450的自然活性，包括P450的伯胺化 苄基和烯丙基C(SP3)-H键通过C-H硝基插入。在这里，我建议进一步扩大这次初选 胺化活性包括天然抗菌剂的底物。我们改造酶的方法是为了 直接胺化C-H键将为加速抗菌药的胺化提供一种手段。精简 抗菌剂的胺化过程反过来又有利于下游的研究，因此这些抗菌剂的抗菌潜力 可以对胺化衍生物进行彻底的研究。这些方法将有助于建立工程化氨基酶。 对于抗菌药，作为概念的证明，可以扩展以帮助加强各种抗菌药。

PROJECT SUMMARY/ABSTRACT Antibiotics are antibacterial drugs used to treat bacterial infections by killing the bacteria or inhibiting bacterial growth to allow immune clearance. While antibiotics help treat infections, their use puts selective pressure on bacteria and inevitably leads to the emergence of resistant bacteria. Additionally, the development of novel antibiotics has slowed in recent years which has left us with fewer treatment options for multidrug-resistant bacterial infections. Moreover, most of these problematic multidrug-resistant bacteria are gram-negative, meaning they are double-membraned and thus intrinsically resistant to many drugs, making drug development even more challenging. There is a dire need for innovative approaches to develop new antibiotics that are effective against gram-negative bacteria so we can preserve our ability to treat these infections. Promising studies have shown that the addition of primary amines can improve drug accumulation into gram-negative bacteria for various compounds, making it a rational and feasible approach to increase the activity of promising antibacterials by improving their bacterial permeation. Synthetically aminating bioactive compounds can be onerous, typically requiring multiple steps and the addition of other functional groups before acquiring the desired amine. In contrast, directed evolution of cytochromes P450 is a promising strategy to develop enzymatic platforms for the late-stage modifications of bioactive compounds. The ability of P450 enzymes to functionalize the inert yet ubiquitous C–H bonds in bioactive compounds can be exploited to facilitate the diversification of natural products. Recent successes have expanded the natural activity of P450s to include primary amination of benzylic and allylic C(sp3) – H bonds via C–H nitrene insertion. Here, I propose to expand further this primary amination activity to include substrates that are natural antibacterials. Our approach to engineering enzymes to directly aminate C–H bonds will provide a means to accelerate the amination of antibacterials. Streamlining the amination process of antibacterials will in turn facilitate downstream studies so the antibacterial potential of these aminated derivatives can be thoroughly investigated. These methods will help establish engineered aminases for antibacterials as a proof of concept that can be expanded to help potentiate various antibacterials.