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

• 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
• 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
• 来源: https://reporter.nih.gov/project-details/10397244
• 关键词: Amination; Amines; Anti-Bacterial Agents; Antibiotics; Bacteria; Bacterial Infections; Binding; Bypass; Chemicals; Chemosensitization; Complex; Cytochrome P450; Development; Directed Molecular Evolution; Drug usage; Effectiveness; Engineering; Enzymes; Escherichia coli; Evolution; Gram-Negative Bacteria; Growth; Heme; Hydrogen Bonding; Infection; Left; Libraries; Membrane; Methods; Modification; Multi-Drug Resistance; Multiple Bacterial Drug Resistance; Mutagenesis; Natural Products; Oxygen; Pharmaceutical Preparations; Process; Reaction; Research; Resistance; Serine; Site; Structure; Terpenes; VDAC1 gene; Variant; Walking; antimicrobial; bacterial resistance; base; drug development; effectiveness study; functional group; immune clearance; improved; in silico; innovation; insight; nitrene; novel; preservation; pressure; screening; success

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.

项目总结/摘要 抗生素是通过杀死细菌或抑制细菌生长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，抗生素的使用速度放缓，这使得我们对多重耐药的治疗选择减少。 细菌感染此外，大多数这些有问题的多重耐药细菌是革兰氏阴性的， 这意味着它们是双膜的，因此本质上对许多药物具有抗性， 更具挑战性迫切需要创新的方法来开发新的抗生素， 对革兰氏阴性菌有效，这样我们就可以保持治疗这些感染的能力。有前途 研究表明，加入伯胺可以改善药物蓄积到革兰氏阴性杆菌中， 细菌对各种化合物的活性，使其成为一种合理可行的途径，以提高活性的有前途的 通过提高其细菌渗透性来抑制细菌。可以合成胺化生物活性化合物， 繁琐的，通常需要多个步骤和添加其他官能团，然后获得所需的 胺·将相比之下，细胞色素P450的定向进化是开发酶的一个有前途的策略， 生物活性化合物后期修饰的平台。P450酶功能化的能力 可以利用生物活性化合物中惰性但普遍存在的C-H键来促进生物活性化合物的多样化。 天然产品。最近的成功已经扩大了P450的天然活性，包括P450的伯胺化。 通过C-H氮烯插入的苄基和烯丙基C（sp3）- H键。在这里，我建议进一步扩大这一主要 胺化活性包括作为天然抗菌剂的底物。我们对酶进行工程改造的方法 直接胺化C-H键将提供一种加速抗菌剂胺化的方法。精简 抗菌剂的胺化过程反过来又会促进下游研究， 胺化衍生物可以被彻底研究。这些方法将有助于建立工程化的氨基酶 作为一个概念的证明，可以扩展到帮助加强各种抗菌药物。