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: 8767927
• 负责人: JOSEPH A POGLIANO
• 金额: $ 68.21万
• 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/8767927
• 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; 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; public health relevance; 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（由申请人提供）：我们开发了一种新方法来促进鉴定具有抗菌活性的天然产物，该方法可以快速区分天然产物 不同的作用机制（MOA）。这种方法称为细菌细胞学分析 (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。针对不同细胞途径和途径内不同步骤的抗生素会产生独特的细胞学特征，从而可以在几个小时内鉴定出新化合物的可能 MOA。我们现在开发了一种补充方法，使我们能够识别抑制目前抗菌药物不靶向的蛋白质的分子。我们将这种方法称为快速抑制分析 (R.I.P.)，它需要快速、诱导性地消耗靶蛋白，然后进行细胞学分析。我们的初步数据表明，R.I.P. 会耗尽药物靶标。产生与相应药物相同的细胞学效应。此外，当前抗菌药物未靶向的必需蛋白质的消耗会产生新的细胞学特征，随后可用于识别抑制这些新靶点的分子。我们在此建议更充分地发展 R.I.P.通过在大肠杆菌和枯草芽孢杆菌的全基因组基础上使用该技术。这将创建一套与抑制基本细胞途径相关的综合参考资料集，这些途径不是当前抗菌药物的目标。全基因组 R.I.P.分析还将提供对保守基因功能的深入了解，我们的初步数据表明，它将提供对协调两个或多个生物合成途径的蛋白质的深入了解，为未来的基础研究提供有趣的起点。然后，我们将使用这个更完整的参考数据集和 BCP 来筛选独特且多样化的天然产物提取物集合，以确定那些抑制这些新药物靶点并杀死多重耐药性的提取物 细菌。然后，我们将与 Medina 基金会的合作者一起纯化和表征我们最优先的先导分子（那些杀死多重耐药细菌的分子），目标是将它们推进毒性试验。