UV-Based Approach for Accessing New Antibiotics

基于紫外线的获取新抗生素的方法

• 批准号: 8469820
• 负责人: Losee Lucy Ling
• 金额: $ 30万
• 依托单位: NOVOBIOTIC PHARMACEUTICALS, LLC
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-06-01 至 2014-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8469820
• 关键词: Actinobacteria class; Animal Model; Anti-Bacterial Agents; Antibiotics; Area; Biological; Cell Line; Chemicals; Chemistry; Code; Collection; Data; Development; Drug resistance; Engineering; Escherichia coli; Fractionation; Genetic Engineering; Genomics; Goals; Growth; Health; Human; In Situ; In Vitro; Infection; Knowledge; Mammalian Cell; Methods; Multi-Drug Resistance; Mutagenesis; Operon; Organism; Pharmaceutical Preparations; Phase; Poison; Production; Pseudomonas aeruginosa; Quinones; Reporting; Resistance; Saccharomyces cerevisiae; Solid; Source; Specificity; Staphylococcus aureus; Streptomyces coelicolor; Structure; Testing; Therapeutic Index; Time; Validation; Vancomycin resistant enterococcus; Work; Yeasts; antimicrobial; bactericide; base; combat; cytotoxicity; cytotoxicity test; drug discovery; genome sequencing; liquid chromatography mass spectrometry; methicillin resistant Staphylococcus aureus; microbial; mutant; novel; novel strategies; pathogen; potency testing; resistant strain; scale up; screening; ultraviolet irradiation

DESCRIPTION (provided by applicant): The goal of this project is to discover novel antibiotics by activating the silent biosynthetic operons of Actinobacteria. Whole genome sequencing of many Actinobacteria showed that there are 10-20 times more operons coding for secondary metabolites than known compounds in a given species. This opens an attractive opportunity to access a large untapped source of new antibiotics. Genetic engineering has been used to turn on silent operons in Actinobacteria, leading to production of secondary metabolites. However, the pace at which engineered operons are activated is very low, less than ten metabolites a year are being reported based on this approach. We reasoned that mutagenesis of isolates that do not produce antimicrobials in vitro will relieve silent operons from regulatory constraints, and screening will then identify the producing mutants. Our preliminary data showed that the approach works surprisingly well, turning over half of the inactive organisms into antibiotic producers. The method is scalable, and we recently identified two potentially novel antimicrobials using this approach. In Phase I, we will mutagenize/screen 2,000 inactive strains for antibiotic production, aiming to obtain new, potentially useful antimicrobials. Biological and chemical dereplication will indicate compounds with potential novelty. We will give priority to broad spectrum compounds with activity against difficult to treat gram-negative pathogens. The antimicrobials will be tested for potency, spectrum, specificity of action and cytotoxicity, and th structure of compounds that pass validation will be determined. Finding 2-3 antimicrobials with novel chemistry will serve as proof-of-principle for this approach. These findings will provide a solid basis for a large-scale drug discovery effort in Phase II.

描述（由申请人提供）：该项目的目的是通过激活静脉细菌的无声生物合成操纵子来发现新型抗生素。许多静脉细菌的整个基因组测序表明，与给定物种中已知的化合物相比，为次级代谢物编码的操纵子多10-20倍。这为获取大量未开发的新抗生素来源开辟了一个诱人的机会。基因工程已被用来打开静脉细菌中的无声操纵子，从而产生次生代谢产物。但是，根据这种方法，每年少于十种代谢产物的速度非常低，每年少于十种代谢产物。我们认为，在体外产生抗菌剂的分离株的诱变将使无声操纵子免于监管的约束，并且 然后，筛选将识别产生的突变体。我们的初步数据表明，该方法效果很好，将一半的无活性生物变成了抗生素生产者。该方法是可扩展的，我们最近使用这种方法确定了两种潜在的新型抗菌剂。在第一阶段，我们将诱变/筛查2,000抗生素生产的非活性菌株，旨在获得新的，潜在的有用的抗菌剂。生物学和化学的消除将表明具有潜在新颖性的化合物。我们将优先考虑具有难以治疗革兰氏阴性病原体的活性的广泛化合物。将测试抗菌素的效力，光谱，作用和细胞毒性的特异性，并确定通过通过验证的化合物的TH结构。发现具有新型化学的2-3个抗菌剂将作为这种方法的原则证明。这些发现将为第二阶段的大规模药物发现工作提供可靠的基础。