Developing Novel Antibiotics Against Yersinia pestis

描述（由申请人提供）：本项目的总体目标是开发针对鼠疫耶尔森氏菌和其他重要人类病原体的广谱抗生素。工程化的多药耐药菌株作为生物恐怖主义的代理人的潜在用途需要开发新的抗Y。鼠疫疗法这是一个具有挑战性的目标，因为最后一类新型广谱抗生素是在20多年前推出的。NovoBiotic拥有能够应对这一挑战的专有技术。我们将从以前无法获得的来源获得新的抗菌剂-不可培养的微生物，占总微生物多样性的99%。在第一阶段，我们的目标是开发一种工业规模的技术，以培养和筛选以前无法培养的微生物用于抗菌素生产。主要的里程碑是“驯化”不可培养的生物体，使其能够在常规生长条件下扩大规模;并开发一种能够以等于或>10（exp 5）分离株/年的速度检测抗菌活性的筛选。我们提前实现了这些里程碑，并拥有强大的技术，使我们能够在第二阶段项目中发现新的抗生素。我们的初步研究结果表明，不可培养的微生物产生新的，广谱的抗菌化合物。我们开发的抗感染药物将瞄准全球240亿美元的市场。广谱抗生素是收入最高的药物，有几种药物的销售额超过10亿美元。第二阶段的具体目标是：1。从不可培养细菌中大规模分离铅抗菌化合物。里程碑：获得200个纯化的潜在新型先导化合物。2.先导化合物的体外验证和结构。a.结构测定。将对来自生物测定指导的分级分离的所有化合物进行结构解析。我们将使用先进的MS和NMR来定义基本的分子结构。B.生物学验证。我们将确定纯化的早期先导化合物的以下性质：对一组测试生物体的效力（MIC）; MBC;在血清存在下的活性;对动物细胞的毒性;抗性发展;作用特异性。里程碑：获得50根结构新颖的电极导线。3.体内验证。通过初步验证的先导化合物将在动物模型中进行详细评价。里程碑：获得3-5个先进的药物开发验证线索。与人类健康的相关性：迫切需要新的抗生素来对付常规和生物战耐药病原体。我们在这个项目中发现的化合物将导致新的广谱抗生素。

DESCRIPTION (provided by applicant): The overall goal of this project is to develop broad-spectrum antibiotics against Yersinia pestis and other important human pathogens. The potential use of engineered, multi-drug-resistant strains as agents of bioterrorism necessitates the development of new anti-Y. pestis therapies. This is a challenging goal, since the last novel class of broad-spectrum antibiotics was introduced over 20 years ago. NovoBiotic has a proprietary technology capable of meeting this challenge. We will obtain new antimicrobials from a previously inaccessible source - unculturable microorganisms that make up 99% of the total microbial diversity. In Phase I, our goal was to develop an industrial-scale technology to grow and screen previously unculturable microorganisms for antimicrobial production. The main milestone was to "domesticate" unculturable organisms to enable scale-up under conventional growth conditions; and developing a screen capable of detecting antimicrobial activity at a rate of equal to or >10(exp5) isolates/year. We achieved these milestones ahead of schedule, and have in place a robust technology that will enable us to discover new antibiotics in this Phase II project. Our preliminary results indicate that unculturable microorganisms produce novel, broad- spectrum antimicrobial compounds. The anti-infectives we develop will target a global $24 billion market. Broad-spectrum antibiotics are the top earners, with several drugs having sales in excess of $1 billion. Phase II Specific Aims are: 1. Large-scale isolation of lead antimicrobial compounds from unculturable bacteria. Milestone: obtain 200 purified, potentially novel lead compounds. 2. In vitro validation and structure of lead compounds. a. Structure determination. All compounds derived from the bioassay guided fractionation will have their structures elucidated. We will define the basic molecular architecture using advanced MS and NMR. b. Biological validation. We will determine the following properties of the purified early leads: potency (MIC) against a panel of test organisms; MBC; activity in the presence of serum; toxicity against animal cells; resistance development; specificity of action. Milestone: obtain 50 leads with novel structure. 3. In vivo validation. The lead compounds that pass preliminary validation will be subjected to a detailed evaluation in an animal model. Milestone: obtain 3-5 advanced validated leads for drug development. Relevance to human health: There is an acute need for new antibiotics to combat conventional and biowarfare drug-resistant pathogens. The compounds we discover in this project will lead to new, broad- spectrum antibiotics.