Identification of natural products targeting new pathways in bacteria

鉴定针对细菌新途径的天然产物

• 批准号: 8848033
• 负责人: JOSEPH A POGLIANO
• 金额: $ 63.56万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-05-12 至 2019-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8848033
• 关键词: Affect; Anti-Bacterial Agents; Antibiotic Therapy; Antibiotics; Bacteria; Basic Science; Biological Factors; Cells; Collection; Data; Data Set; Drug Targeting; Drug resistance; Escherichia coli; Fluorescence Microscopy; Future; Genes; Goals; Gram-Negative Bacteria; Gram-Positive Bacteria; Health; Hour; Individual; Lead; Measures; Multi-Drug Resistance; Organism; Pathway interactions; Peptide Hydrolases; Pharmaceutical Preparations; Proteins; Research; Resistance; Technology; Toxic effect; Wit; base; cellular targeting; combat; genome-wide; improved; insight; interest; killings; novel; novel strategies; pathogen; protein profiling; safety testing

DESCRIPTION (provided by applicant): We developed a new approach to facilitate identification of natural products with antibacterial activities that rapidly discriminates between different mechanisms of action (MOA). This approach, bacterial cytological profiling (BCP), uses quantitative fluorescence microscopy to measure the effects of antibiotic treatment on individual cells. Antibiotics that target different cellular pathways and different steps within a pathway generate unique cytological profiles, allowing identification of the likely MOA of new compounds within a few hours. We have now developed a complimentary approach that will allow us to identify molecules that inhibit proteins that are not currently targeted by antibacterial drugs. Ths approach, which we call rapid inhibition profiling (R.I.P.), entails the rapid, inducible depletionof a target protein, followed by cytological profiling. Our preliminary data demonstrate that depletion of a drug target by R.I.P. produces cytological effects identical to those produced by the corresponding drug. Furthermore, depletion of essential proteins that are not targeted by current antibacterial drugs produces novel cytological profiles that can be subsequently used to identify molecules that inhibit these new targets. We here propose to more fully develop the R.I.P. technology by employing it on a genome wide basis in E. coli and B. subtilis. This will create a comprehensive reference set of profiles associated with the inhibition of essential cellular pathways that are not the targets of current antibacterial drugs. The genome-wide R.I.P. analysis will also provide insight into the function of conserved genes and our preliminary data suggests that it will provide insight into proteins that coordinate two or more biosynthetic pathways, providing interesting starting points for future basic research. We will then use this more complete reference data set with BCP to screen a unique and diverse collection of natural product extracts to identify those that inhibit these new drug targets and kill multidrug resistant bacteria. Together with our collaborators at Fundaci¿n Medina, we will then purify and characterize our highest priority lead molecules (those that kill multidrug resistant bacteria) wit a goal of advancing them into toxicity trials.

描述（由应用程序提供）：我们开发了一种新的方法，以促进具有抗菌活动的天然产品，以迅速区分 不同的作用机理（MOA）。这种方法是细菌细胞学分析（BCP），使用定量荧光显微镜来测量抗生素治疗对单个细胞的影响。靶向不同细胞途径和不同步骤的抗生素会产生独特的细胞学特征，从而在几个小时内鉴定了可能的新化合物MOA。现在，我们已经开发了一种免费的方法，该方法将使我们能够鉴定出抑制抗菌药物当前靶向的蛋白质的分子。我们称之为快速抑制分析（R.I.P.）的方法需要对靶蛋白的快速，可诱导的耗竭，然后进行细胞学分析。我们的初步数据表明，R.I.P.将药物靶向部署产生与相应药物产生的细胞学作用相同。此外，当前抗菌药物未靶向的必需蛋白质的部署会产生新的细胞学特征，随后可用于鉴定抑制这些新靶标的分子。我们在这里提议更充分地开发R.I.P.通过在大肠杆菌和枯草芽孢杆菌中以广泛的基因组为基因组来使用技术。这将创建与抑制不是当前抗菌药物靶标的必需细胞途径相关的全面参考。全基因组R.I.P.分析还将提供对保守基因功能的洞察力，我们的初步数据表明，它将提供对蛋白质协调两个或多个生物合成途径的洞察力，为未来的基础研究提供了有趣的起点。然后，我们将使用BCP使用此更完整的参考数据集来筛选独特而多样化的天然产品提取物，以识别那些抑制这些新药物的靶标并杀死耐多药物的靶标。 细菌。然后，我们将与我们在Fundaci n Medina的合作者一起进行净化并表征我们最高优先级的铅分子（杀死多药细菌的分子），这是将它们推进到毒性试验中的目标。