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

用于生物活性分子构建和多样化的非生物酶 C-H 功能化

基本信息

批准号：
10397244
负责人：
FRANCES H ARNOLD
金额：
$ 6.89万
依托单位：
CALIFORNIA INSTITUTE OF TECHNOLOGY
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-07-01 至 2024-06-30
项目状态：
已结题

项目摘要

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.

项目摘要/摘要抗生素是用于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全天候清除。抗生素有助于治疗感染，但它们的使用构成选择性压力细菌和不可避免地会导致抗性细菌的出现。另外，新颖的发展近年来，抗生素已经放缓，这使我们的治疗选择更少细菌感染。此外，大多数这些有问题的耐多药细菌都是革兰氏阴性菌，这意味着它们是双重的，因此对许多药物具有本质上的抵抗力，从而使药物开发更多的挑战。迫切需要创新的方法来开发新的抗生素有效防止革兰氏阴性细菌，因此我们可以保留治疗这些感染的能力。有希望的研究表明，添加原代胺可以改善药物的积累到革兰氏阴性中各种化合物的细菌，使其成为增加承诺活动的合理和可行方法通过改善细菌渗透的抗菌作用。合成修订的生物活性化合物可以是繁重的，通常需要多个步骤，并在加速所需的其他功能组之前添加其他官能团胺。相反，细胞色素P450的定向演变是开发酶促的有前途的策略生物活性化合物晚期修饰的平台。 P450酶功能化的能力可以探索生物活性化合物中无处不在的C – H键，以促进多样化天然产品。最近的成功扩大了P450的自然活动，包括通过C – H硝基插入的苯基和Allic C（SP3） - H键。在这里，我建议进一步扩展这一主要胺化活性包括天然抗菌物的底物。我们的工程酶方法直接成功的C – H债券将提供一种加速抗菌胺化的方法。简化抗菌物的胺化过程反过来将紧急下游研究，因此这些抗菌潜力可以彻底研究布置的衍生物。这些方法将有助于建立工程amase 对于抗菌作为概念证明，可以扩展以帮助潜在的各种抗菌物。