Probing a novel allosteric binding site in Mycobacterium DNA gyrase to tackle TB and antimicrobial resistance

探索分枝杆菌 DNA 旋转酶中的新型变构结合位点以应对结核病和抗菌素耐药性

• 批准号: 2739699
• 金额: --
• 依托单位: University of Leeds
• 依托单位国家: 英国
• 项目类别: Studentship
• 财政年份: 2022
• 资助国家: 英国
• 起止时间: 2022 至 无数据
• 项目状态: 未结题
• 来源: https://gtr.ukri.org/projects?ref=studentship-2739699
• 关键词: Probing; novel; allosteric; binding; site

BackgroundAntimicrobial resistance (AMR) undermines the effectiveness of antibiotics. Fluoroquinolones are frontline antibiotics that treat bacterial infections via the inhibition of DNA gyrase. A recently characterised allosteric site on Staphylococcus aureus DNA gyrase offers the potential for the design of allosteric inhibitors of gyrase. Since the allosteric site is distinct from the fluoroquinolone-binding site, allosteric inhibitors can overcome fluoroquinolone resistance observed in the clinic (point mutations in gyrase). The project aims to investigate this allosteric site in Mycobacterium tuberculosis (Mtb) which has been unexplored to date, but has high amino acid conservation compared to the sequence of S. aureus gyrase.Objectives1. Use computational techniques and homology modelling to build a structural model of the Mtb allosteric site.2. Use computational medicinal chemistry to design potent inhibitors of Mtb.3. Ascertain the mode of action of the allosteric inhibitors: do they bind to the nicked or cleaved DNA complex?4. Establish synergy of allosteric inhibitors with fluoroquinolones.Novelty1.4 million people died of TB in 2019 and bovine TB is a major infectious disease in cattle. Multi-drug resistance (XDR)-TB are strains resistance to nearly all frontline antibiotics. Therefore, new investigations into viable therapeutic targets is required such as this allosteric site.TimelinessThis work capitalises on our recent discovery of Escherichia coli gyrase allosteric inhibitors with cellular activity against Mtb (MIC90 8 uM). The supervisory team have an established track record in investigating small molecule inhibition of bacterial DNA gyrase. All the methodologies for these studies have been established.Experimental ApproachThe student will be trained in computational medicinal chemistry (to understand protein structure, ligand-protein interactions, computer-aided drug design) and synthetic chemistry. Synthesised molecules will be characterized using a combination of biochemistry and crystallography to ascertain their mode of Mtb gyrase inhibition.

背景抗微生物耐药性（AMR）破坏了抗生素的有效性。氟喹诺酮是前线抗生素，可通过抑制DNA回旋酶治疗细菌感染。最近在金黄色葡萄球菌DNA回旋酶上的变构位点的变构位点提供了设计陀螺酶变构抑制剂的潜力。由于变构位点与氟喹诺酮结合位点不同，因此变构抑制剂可以克服诊所中观察到的氟喹诺酮耐药性（Gyrase中的点突变）。该项目旨在研究结核分枝杆菌（MTB）中的这种变构位点，迄今为止尚未探索，但与金黄色葡萄球菌gylase.Objectives的序列相比，氨基酸保护很高。使用计算技术和同源性建模来构建MTB变构位点的结构模型2。使用计算药物设计MTB.3的有效抑制剂。确定变构抑制剂的作用方式：它们是否结合到划痕或裂解的DNA复合物？4。建立与氟喹诺酮类药物的变构抑制剂的协同作用。2019年，有140万人死于结核病，牛结核病是牛的主要传染病。多药耐药性（XDR）-TB是对几乎所有前线抗生素的抗性抗性。因此，需要对可行的治疗靶标进行新的研究，例如这种变构位点。Timelinesstimeliness这可能会利用我们最近发现具有针对MTB的细胞活性的大肠杆菌回旋酶变构抑制剂（MIC90 8 UM）。监督小组在研究细菌DNA循环酶的小分子抑制方面具有既定的记录。这些研究的所有方法都已建立。实验方法将接受计算药物化学（了解蛋白质结构，配体 - 蛋白质相互作用，计算机辅助药物设计）和合成化学的培训。合成分子将使用生物化学和晶体学的组合来确定其MTB回旋酶抑制模式。