TUsing NMR to enable the development and derive the mode of action of novel antimicrobial technologies

该项目将开发一种新的、综合的细胞内方法，利用溶液核磁共振(核磁共振)光谱的原子细节，以应对目前限制抗菌药物开发的挑战。超分子自缔合两亲化合物(SSA)是JH发明的新型100种表面活性抗菌剂和抗生素/杀生剂增效剂。项目目标：开发一套使用生物反应器和核磁共振谱仪的新型高通量方法，使我们能够在原子水平上推导出这项技术创新的作用机制。这些研究将通过使用免费的纳米光盘研究(参考文献3和4)和针对多重耐药ESKAPE病原体(CH)的微生物学技术来验证。目的：1.优化监测SSA内化的核磁共振方法。2.检测与细菌相互作用时的分子结构变化。3.监测细菌在抗菌剂存在下的代谢变化。4.代谢水平上抗性机制的确定。5.根据产生的实验数据，开发具有增强的抗菌剂/功效增强特性的下一代SSA。JOR实验室：开发/优化细胞内的核磁共振分析(目标1、3、4)。测试初始组SSA分子，以阐明它们的流入/流出速率和对细胞的影响(目标1、3)。测试对SSA的潜在抗性机制(目标4)。JH实验室：合成和设计迭代生成的SSA(目标2-5)。NW实验室：阐明SSA与生物膜相互作用时的结构变化(目标2)。CH实验室：测定SSA对多药耐药ESKAPE病原体的疗效(目标5)。影响领域：1.更健康的抗菌治疗/商业去污产品。2.生物经济--SSA技术商业化。3.人员和人才--对STEM内妇女和其他边缘化群体的支持--详情见后文各节。

This project will develop a new, integrative, in-cell methodology exploiting the atomic detail of solution Nuclear Magnetic Resonance (NMR) spectroscopy, to tackle the challenges which currently limit antimicrobial development. Supramolecular Self-associating Amphiphiles (SSAs) are a new class of 100 surface active antimicrobial agents and antibiotic/biocide efficacy enhancers invented by JH. Project aim: To develop a suite of novel high-throughput methods using a bioreactor coupled to an NMR spectrometer, that will enable us to derive the mechanism of action for this technological innovation on the atomic level. These studies will be validated through the use of complimentary nanodisc studies (references 3 and 4) and microbiological techniques against multidrug resistant ESKAPE pathogens (CH). Objectives: 1. Optimise NMR methods to monitor SSA internalisation. 2. Detection of molecular structural alterations upon interaction with bacteria. 3. Monitoring metabolic changes of the bacteria in the presence of antimicrobial agents. 4. Determination of resistance mechanisms at the metabolic level. 5. Development of next-generation SSAs with enhanced antimicrobial/efficacy enhancement properties based on the experimental data generated. JOR lab: Develop/optimize NMR assays in cells (Objectives 1, 3,4).Test initial set of SSA molecules to elucidate their influx/efflux rates and effect on cells (Objectives 1, 3). Test potential resistance mechanisms against SSAs (Objective 4). JH Lab: Synthesis and design of iterative generations of SSAs (Objectives 2-5). NW Lab: Elucidation of the structural changes of SSAs upon interaction with biological membranes (Objective 2). CH Lab: Determination of SSA efficacy against multidrug resistant ESKAPE pathogens (Objective 5). Impact areas: 1. Health - better antimicrobial treatments/commercial decontamination products. 2. Bioeconomy - Commercialisation of SSA technology. 3. People and talent - Support of women and other marginalised groups within STEM - See later sections for detail.