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、细胞内的方法，利用溶液核磁共振（NMR）光谱的原子细节，以应对目前限制抗菌剂开发的挑战。超分子自缔合两亲物（SSA）是由JH发明的一类新的100种表面活性抗微生物剂和抗生素/杀生物剂功效增强剂。 项目目标：为了开发一套新的高通量方法，使用生物反应器耦合到NMR光谱仪，这将使我们能够推导出原子水平上这项技术创新的作用机制。 这些研究将通过使用针对多药耐药ESKAPE病原体（CH）的补充纳米片研究（参考文献3和4）和微生物学技术进行验证。 目的：1.优化NMR方法以监测SSA内化。2.检测与细菌相互作用后的分子结构改变。3.在抗菌剂存在下监测细菌的代谢变化。4.在代谢水平上确定耐药机制。5.基于生成的实验数据开发具有增强的抗菌/功效增强特性的下一代SSA。 JOR实验室：开发/优化细胞中的NMR分析（目标1，3，4）。测试初始SSA分子组以阐明其流入/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：SSA迭代生成的合成和设计（目标2-5）。 NW实验室：阐明SSA与生物膜相互作用后的结构变化（目标2）。 CH实验室：确定SSA对多药耐药ESKAPE病原体的疗效（目标5）。 影响范围：1。健康-更好的抗菌处理/商业去污产品。2.生物经济-SSA技术的商业化。3.人员和人才-支持STEM中的妇女和其他边缘化群体-详见下文。

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.