DEVELOPMENT OF A NOVEL CLASS OF ANTIBIOTICS AGAINST VIBRIO CHOLERAE NA+-NQR

新型抗霍乱弧菌 NA -NQR 抗生素的开发

• 批准号: 10540361
• 负责人: Oscar Juarez
• 金额: $ 55.71万
• 依托单位: ILLINOIS INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-12-13 至 2025-11-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10540361
• 关键词: Animal Model; Antibiotics; Antimalarials; Antipsychotic Agents; Automobile Driving; Bacterial Infections; Binding; Binding Sites; Biochemistry; Biological; Cell physiology; Cells; Cellular biology; Cholera; Communicable Diseases; Complex; Computer Analysis; Coupled; Crystallization; Data; Development; Diarrhea; Docking; Dose; Drug Design; Drug Efflux; Drug Targeting; Enzyme Inhibitor Drugs; Enzymes; Equilibrium; Evolution; Family; Foundations; Free Energy; Gastrointestinal Diseases; Generations; Growth; Human; Human Genome; Immune system; Infection; Institution; Intestines; Ions; Lead; Libraries; Life; Malaria; Mammalian Cell; Membrane; Metabolism; Methods; Microbe; Microbiology; Mitochondria; Modeling; Multi-Drug Resistance; Multienzyme Complexes; Multiple Bacterial Drug Resistance; NADH; Na(+)-K(+)-Exchanging ATPase; Nutrient; Oxidation-Reduction; Oxidoreductase; Pathogenicity; Pattern; Performance; Persons; Pharmaceutical Chemistry; Pharmaceutical Preparations; Phenothiazines; Phenotype; Physiology; Play; Predisposition; Process; Production; Property; Proteins; Psychoses; Reactive Oxygen Species; Regulation; Role; Rotation; Site; Sodium; Solid; Specificity; Structure; Testing; Toxic effect; Toxin; Ubiquinone; Vibrio cholerae; analog; bacterial metabolism; cell motility; design; drug development; drug-like compound; functional group; gastrointestinal infection; human disease; improved; in vivo; inhibitor; iterative design; lead optimization; microbial; microorganism; mutant; novel; novel antibiotic class; novel therapeutics; pandemic disease; pathogen; pathogenic bacteria; pharmacologic; pharmacophore; process optimization; resistance mechanism; resistant strain; respiratory; respiratory enzyme; scaffold; structural biology; uptake

PROJECT SUMMARY The sodium-dependent NADH: ubiquinone oxidoreductase (Na+-NQR) is the main ion transporter in hundreds of pathogenic bacteria, including Vibrio cholerae, the causal agent of cholera, a devastating gastrointestinal disease with a worldwide distribution that has developed multidrug-resistant phenotypes. Na+- NQR fulfills two essential roles in V. cholerae cell physiology, as a respiratory enzyme, providing energy to the cell, and as the main sodium pump, energizing the membrane and driving nutrient uptake, pH regulation, elimination of drugs, cell motility, secretion of toxins and other homeostatic processes. Na+-NQR is an optimal drug target due to its critical role in bacterial metabolism and because it is absent in mammalian cells. Moreover, Na+-NQR has unique structural motifs, not found in any human protein, which allow the discovery of drugs that can act specifically on this enzyme. In addition, Na+-NQR inhibitors could increase the susceptibility of V. cholerae to other drugs by de-energizing the membrane, and may be used in a combination dosing approach to rescue obsolete antibiotics. Our group has now identified two novel compound leads, ubiquinone analogs (UQAs) and phenothiazines, as inhibitors of this enzyme that are suitable for drug development. The three UQAs analogs characterized have antimalarial properties and show specific and potent inhibitory effects on Na+-NQR, with strong antibiotic activity against V. cholerae. These compounds not only abolish V. cholerae Na+-NQR enzymatic activity, but also trigger the overproduction of reactive oxygen species, which is lethal to microbes. The structures of these inhibitors and docking methods were used to identify the pharmacophore and the binding modes of the molecules in the UQ binding site, which allow us to pursue lead development to obtain inhibitors of high potency and specificity. In addition, we have identified three phenothiazine-like compounds with anti- psychotic properties that show potent inhibitory activity against Na+-NQR and that could be optimized into antibiotics. The main aim of this project is the development of a novel class of antibiotics to specifically target the Na+-NQR complex. The inhibitors that we have identified will be fully characterized, to understand their mechanism of action, binding sites, potency and antibiotic properties. Moreover, toxicity towards human cells and mitochondria, as well as their pharmacologic properties, will be assessed to evaluate the potential of these compounds to treat human infections. The data obtained from toxicity studies, enzymatic and microbiological characterizations will be used to guide the design and synthesis of analogs with high potency and low toxicity. Lead optimization will be carried out by our medicinal chemistry team guided by pharmacophore analysis, docking and binding free energy calculations. The structures of the newly-identified inhibitors will be used to build compound libraries carrying the active core with different substitution patterns, which will be iteratively screened and characterized. The data generated in this proposal is critical to the discovery of urgently-needed antibiotics with a new mechanism of action effective against V. cholerae and many other Na+-NQR bearing pathogens.

项目摘要 依赖钠的NADH：泛酮酮氧化还原酶（Na+-NQR）是主要离子转运蛋白 数百种致病细菌，包括霍乱的弧菌霍乱，霍乱的因果剂，一种毁灭性的 胃肠道疾病具有全球分布，已经发展出多药耐药性表型。 Na+ - NQR在霍乱细胞生理学中扮演两个基本角色，作为一种呼吸酶，为能量提供了能量 细胞，作为主钠泵，使膜充满活力并驱动养分吸收，pH调节， 消除药物，细胞运动，毒素的分泌和其他稳态过程。 Na+-NQR是最佳的 药物靶标由于其在细菌代谢中的关键作用而引起的，并且由于它在哺乳动物细胞中不存在。而且， Na+-NQR具有独特的结构基序，在任何人类蛋白质中都没有发现，这允许发现药物 可以专门针对该酶。此外，Na+-NQR抑制剂可能会增加V的敏感性。 通过脱氧膜与其他药物的霍乱，可用于组合剂量的方法 营救过时的抗生素。我们的小组现在已经确定了两个新型的化合物铅泛酮类似物 （UQAS）和苯噻嗪作为适合药物开发的酶的抑制剂。三个uqas 表征的类似物具有抗疟疾特性，并对Na+-NQR显示出特异性和有效的抑制作用， 具有强对霍乱的抗生素活性。这些化合物不仅废除了V.霍乱na+-nqr 酶促活性，但也会触发活性氧的过量生产，这对微生物致命。 这些抑制剂和对接方法的结构用于鉴定药效团和结合 UQ结合位点中分子的模式，这使我们能够追求铅开发以获得抑制剂 高效力和特异性。此外，我们已经确定了三种势噻嗪样化合物 具有对Na+-NQR的有效抑制活性的精神病特性，可以将其优化到 抗生素。该项目的主要目的是开发一种新型的抗生素，以专门针对 Na+-NQR复合物。我们确定的抑制剂将被充分表征，以了解他们 作用机理，结合位点，效力和抗生素特性。此外，对人类细胞的毒性 将评估线粒体及其药理特性，以评估这些潜力 治疗人类感染的化合物。从毒性研究，酶和微生物学获得的数据 特征将用于指导高效力和低毒性的类似物的设计和合成。 铅优化将由我们的药化化学团队在药丸分析指导下进行， 对接和结合自由能计算。新识别的抑制剂的结构将用于建造 带有不同替代模式的活动核心的复合库，将迭代筛选 并描述了。本提案中产生的数据对于发现急需的抗生素至关重要 具有针对V.霍乱和许多其他NA+-NQR病原体的新作用机理。