Tools and Reagents for Next Generation Inhibitor Discovery in Peptidoglycan biosynthesis

对现有抗生素的抗药性在多个层面上威胁着未来的医疗保健，并已被公认为一个世界性问题，其影响与气候变化一样重要。然而，尽管存在明显的临床需求，但一些因素导致制药业抗菌药的发现和开发稳步下降。剩余的制药公司在这一领域的抗菌研究和开发过去通常侧重于尝试发现抗生素的新靶点，或基于现有靶点发现活性有限的新化合物。一般来说，这种方法取得的成功非常有限，再加上其他因素，这意味着我们治疗细菌感染的药物数量减少，导致医疗保健危机。显然，细菌擅长针对单基因靶点(即其蛋白质产物)选择抗药性，过去成功的抗微生物化疗选择了具有多种基本活动的生物过程，例如核糖体上的蛋白质合成，负责细胞内DNA超级卷曲的蛋白质，以及实现细菌细胞壁聚合物的形成和交联的机制。细菌细胞壁聚合物的生物合成称为肽聚糖，是这方面抗微生物药物的极佳靶点，因为它提供了如上所述的多靶向机会，而不是在动物或人类中发现的结构，因此不太可能产生不利的化学敏感性，并且该过程发生在细胞外，允许药物分子更容易进入。虽然青霉素类药物过去曾被用来利用这些优势，但它们的目标是相关细菌酶的交联活性。这些酶，称为青霉素结合蛋白，也负责在一个完全不同的反应中形成肽聚糖聚合物，到目前为止，该反应一直被视为抗生素的靶标。我们的提案旨在获得为下一代抗生素探索这些酶所需的基本知识。我们的建议将学术界和工业界的知识和进步联系在一起，共同努力获取所需的关键信息。至关重要的是，我们需要产生一系列专业的化学探针来询问这些酶的工作机制，确定这些探针与酶结合的结构，以产生新的分析方法，从而能够发现未来的抗生素。

Antimicrobial resistance to existing antibiotics threatens future healthcare at multiple levels and has been acknowledged as a worldwide issue with an impact as important as climate change. However, a number of factors has led to a steady decline in the discovery and development of antimicrobials in the pharmaceutical industry despite the clear clinical need. The remaining pharmaceutical company antibacterial research and development in this area has focused generally in the past on either attempts to discover new targets for antibiotics or new compounds with limited activity profiles based on existing targets. Generally this approach has met with very limited success which combined with other factors, means we have a decreasing number of drugs to treat bacterial infection leading to a crisis in healthcare. It is clear that bacteria are adept at the selection of resistance to drugs for single gene targets (i.e. their protein products) and that the successful antimicrobial chemotherapy of the past, selects biological processes where there are multiple essential activities e.g. protein synthesis at the ribosome, the proteins which are responsible for DNA supercoiling in the cell and mechanisms by which formation and cross-linking of the bacterial cell wall polymer is achieved. The biosynthesis of the bacterial cell wall polymer called peptidoglycan is an excellent target for antimicrobials in this respect since it provides an opportunity for multi-targeting as described above, is not a structure found in animals or humans so there is less chance of adverse chemical sensitivity and the process occurs outside the cell allowing easier access for drug molecules. Whilst the penicillin group of drugs have been used to exploit these advantages in the past, they target the cross-linking activity of the bacterial enzymes concerned. These enzymes, called penicillin binding proteins, are also responsible for the formation of the peptidoglycan polymer as well in a completely separate reaction that has hitherto been under explored as a target for antibiotics.Our proposal seeks to gain the essential knowledge required to explore these enzymes for next generation antibiotics. Our proposal links the knowledge and advances in academia and industry in a mutually beneficial effort to get the key information required. Critically, we need to generate a series of specialist chemical probes to interrogate the mechanism by which these enzymes work, to determine the structures of these probes bound to the enzymes to generate new assays that will allow the discovery of future antibiotics.