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，来解决目前限制抗菌药物开发的挑战。超分子自缔合两亲体（SSAs）是JH公司发明的一类具有100种表面活性的抗菌剂和抗菌/杀菌剂功效增强剂。项目目标：开发一套新的高通量方法，使用生物反应器与核磁共振光谱仪相结合，这将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互补的纳米盘研究（参考文献3和4）和针对多药耐药ESKAPE病原体（CH）的微生物学技术来验证。目的:1。优化核磁共振方法以监测SSA内部化。2. 与细菌相互作用时分子结构变化的检测。3. 监测抗菌药物存在下细菌的代谢变化。4. 代谢水平耐药机制的测定。5. 基于实验数据开发具有增强抗菌/功效增强特性的下一代SSAs。主要实验室：开发/优化细胞核磁共振分析（目标1、3、4）。测试初始组SSA分子以阐明其内流/流出速率和对细胞的影响（目的1,3）。测试SSAs的潜在耐药机制（目的4）。JH实验室：SSAs迭代代的合成和设计（目标2-5）。NW实验室：阐明SSAs与生物膜相互作用时的结构变化（目标2）。实验室：测定SSA对ESKAPE多药耐药病原菌的疗效（目的5）。影响范围：1；健康——更好的抗菌治疗/商业去污产品。2. 生物经济- SSA技术的商业化。3. 人员和人才-在STEM中支持妇女和其他边缘化群体-详见后面的章节。

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.