Developing Novel Antibiotics Against Yersinia pestis

描述(由研究人员提供)：该项目的总体目标是开发针对鼠疫耶尔森氏菌的治疗方法，鼠疫耶尔森氏菌是鼠疫的病原体，也是最危险的潜在生物武器之一。我们最近开发了一种通用的方法来培养以前“不可培养”的微生物(Kaeberlein，T.，Lewis，K.和Epstein，S.S.(2002))。Science 296：1127-1129)，这项专利技术将作为抗生素发现的平台。目前使用的大多数抗生素来自可培养微生物，占所有物种的1%。由于对可栽培物种的过度开采，新化合物的发现在70年代基本上停止了。不可培养的微生物是地球上最丰富的生物多样性来源，我们将获得这一基本上无限的资源，以发现对抗鼠疫杆菌的抗生素。第一阶段的目标是从不可培养的微生物中筛选出抗菌化合物，并通过鉴定具有化学新颖性的化合物来获得这项技术的原理证明。这将使第二阶段的大规模筛选和验证成为可能，这将产生先导化合物，最终产生一种新的抗生素产品。 具体目标是： 1.获取和筛选不可培养微生物的工具。扩散箱法将用于分离和培养不可培养的微生物。(里程碑：我们将通过这种方法获得1000个新的分离物)。将开发其他快速分离、筛选和扩大规模的新方法：将确定辅助微生物，使不可培养的微生物能够在体外合成介质上生长。驯化不能培养的微生物将导致在常规合成培养基上生长出不同的微生物。(里程碑：使用这些方法，我们将实现每年对105株抗生素生产商进行筛查的速度)。 2.提取液筛选及初步验证。(里程碑：筛选4000个摘录)。 3.去复制和结构测定。(里程碑：确定5种化学新化合物)。

DESCRIPTION (provided by investigator): The overall goal of this project is to develop therapies against Yersinia pestis, the causative agent of plague and one of the most dangerous potential bioweapons. We recently developed a general method to grow previously "unculturable" microorganisms (Kaeberlein, T., Lewis, K., and Epstein, S.S. (2002). Science 296:1127-1129), and this proprietary technology will serve as a platform for antibiotic discovery. Most antibiotics currently in use are derived from cultivable microorganisms that make up <1% of all species. Discovery of novel compounds essentially ceased in the 70s due to overmining of cultivable species. Unculturable microorganisms are the richest source of biodiversity on this planet, and we will access this essentially unlimited resource to discover antibiotics acting against Y. pestis. The goal of Phase I is to establish screens for antimicrobial compounds from unculturable microorganisms, and obtain proof-of principle for this technology by identifying compounds with chemical novelty. This will enable large-scale screening and validation in Phase II that will produce lead compounds, ultimately resulting in a new antibiotic product. The Specific Aims are: 1. Tools to obtain and screen unculturable microorganisms. The diffusion chamber approach will be used to isolate and grow unculturable microorganisms. (Milestone: we will obtain 1,000 novel isolates by this method). Additional novel methods for rapid isolation, screening and scale up will be developed: Helper organisms will be identified that enable growth of unculturable microorganisms on synthetic media in vitro. Domestication of unculturable microorganisms will result in variant growing on regular synthetic media. (Milestone: using these methods, we will achieve a rate of screening for antibiotic producers of >105 isolates/year). 2. Extract screening and preliminary validation. (Milestone: screen 4,000 extracts). 3. Dereplication and structure determination. (Milestone: identify 5 chemically novel compounds).