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）：该项目的目标是通过激活放线菌的沉默生物合成操纵子来发现新型抗生素。许多放线菌的全基因组测序表明，在给定物种中，编码次级代谢产物的操纵子比已知化合物多 10-20 倍。这为获取大量未开发的新型抗生素提供了一个诱人的机会。基因工程已被用来打开放线菌中的沉默操纵子，从而产生次级代谢产物。然而，工程操纵子被激活的速度非常低，基于这种方法每年报告的代谢物不到十种。我们推断，对体外不产生抗菌剂的分离株进行诱变将解除沉默操纵子的监管限制，并且 然后筛选将鉴定产生突变体。我们的初步数据表明，这种方法效果出人意料地好，将一半以上的不活跃生物转化为抗生素生产者。该方法具有可扩展性，我们最近使用这种方法发现了两种潜在的新型抗菌剂。在第一阶段，我们将诱变/筛选 2,000 个用于抗生素生产的无活性菌株，旨在获得新的、潜在有用的抗菌药物。生物和化学去重复将表明化合物具有潜在的新颖性。我们将优先考虑对难以治疗的革兰氏阴性病原体具有活性的广谱化合物。将测试抗菌剂的效力、谱、作用特异性和细胞毒性，并确定通过验证的化合物的结构。寻找 2-3 种具有新颖化学成分的抗菌剂将作为该方法的原理验证。这些发现将为第二阶段的大规模药物发现工作提供坚实的基础。