Narrow-Spectrum Drug Targets for Bacillus anthracis

炭疽杆菌的窄谱药物靶点

• 批准号: 6669929
• 负责人: William W Barrow
• 金额: $ 22.77万
• 依托单位: OKLAHOMA STATE UNIVERSITY STILLWATER
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-09-30 至 2005-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6669929
• 关键词: Bacillus anthracis; X ray crystallography; antibacterial agents; bioterrorism /chemical warfare; chemical registry /resource; dihydrofolate reductase; drug design /synthesis /production; drug discovery /isolation; drug screening /evaluation; enzyme mechanism; high throughput technology; ligands; molecular cloning; other phosphotransferase; polymerase chain reaction; protein sequence; protein structure function; recombinant proteins; transaldolase /transketolase

DESCRIPTION (provided by investigator): This application is in response to NIAID's request for high-priority biodefense products specific to CDC Category A-C Priority Pathogens. It focuses on the development of compounds with selective activity against Bacillus anthracis, included in the highest priority group, Category A. The goal of this application is to show proof-of-principal for potential antimicrobial targets in a specific biosynthetic pathway of Bacillus anthracis that will allow for the development of compounds that can be used to treat anthrax. At least two enzymes in this pathway represent targets that are naturally resistant to standard antimicrobial agents. As a result, traditional drugs for these targets are currently not recommended for treatment of anthrax. We hypothesize that these targets are amenable to new drug development and that they may be susceptible to non-traditional antimicrobials that may already exist in chemical repositories. Consequently, one goal of this project will be to implement the most expedient way to screen drugs that are already available against these enzyme targets. Because the degree of homology of these enzymes is so high, we hypothesize that B. cereus can be used in place of Bacillus anthracis. This will facilitate the use of high-throughput screening with a robotics system in a BSL-2 versus a BSL-3 facility. The approach will be to use DNA recombinant technology coupled with X-ray crystallographic molecular modeling to demonstrate proof-of-principal with regard to functional similarities of two key enzymes in an essential metabolic pathway of Bacillus anthracis. Recombinant enzymes will be used in drug-screening assays and for crystallization to develop effective three-dimensional models to confirm enzyme similarities. If successful, this strategy would prove useful for other enzymes in this pathway that also share a high degree of homology. This R21 application is intended to lay the groundwork for a subsequent RO1 that will be designed to attain narrow-spectrum antibiotics for Bacillus anthracis through early product development resulting from collaborations between academic researchers and industrial laboratories.

描述（由研究者提供）：本申请是对NIAID针对CDC A-C类优先病原体的高优先级生物防御产品的要求的回应。它侧重于开发对炭疽杆菌具有选择性活性的化合物，包括在最高优先级组A类中。本申请的目的是显示炭疽芽孢杆菌特定生物合成途径中潜在抗菌靶标的原理证明，从而开发可用于治疗炭疽的化合物。该途径中至少有两种酶代表天然耐标准抗菌剂的靶标。因此，针对这些目标的传统药物目前不推荐用于治疗炭疽。我们假设，这些目标是服从新药开发，他们可能是易受非传统的抗菌剂，可能已经存在于化学仓库。因此，该项目的一个目标将是实施最便捷的方法来筛选针对这些酶靶点的药物。由于这些酶的同源性程度如此之高，我们假设B。蜡状芽孢杆菌可用于代替炭疽芽孢杆菌。这将有助于在BSL-2与BSL-3设施中使用机器人系统进行高通量筛选。该方法将使用DNA重组技术结合X射线晶体学分子建模来证明炭疽芽孢杆菌基本代谢途径中两种关键酶的功能相似性的原理证明。重组酶将用于药物筛选试验和结晶，以开发有效的三维模型来确认酶的相似性。如果成功的话，这种策略将证明对该途径中也具有高度同源性的其他酶有用。该R21应用旨在为随后的RO 1奠定基础，该RO 1旨在通过学术研究人员和工业实验室之间的合作进行早期产品开发，以获得针对炭疽杆菌的窄谱抗生素。