Antibiotic Drug Discovery

抗生素发现项目结合了化学和生物学，以确定新的化合物的效用， 治疗或预防由革兰氏阴性菌引起的疾病。该项目开发高吞吐量 筛选，执行化学合成作为命中的一部分，以导致化学，测量生物效应， 用纯化的酶和全菌进行复合，并研究复合物在小动物中的功效 疾病的模型。目前有三个独立的研究项目：（1）革兰氏阴性毒力蛋白 噻唑烷酮的分泌抑制（2）多种策略，包括类似物的化学合成 和高通量筛选开发，以发现抑制三个步骤的化合物， 第二信使di-c-GMP代谢和（3）测试不具有内在抗生素的喹诺酮 活性，但促进阳离子抗生素多粘菌素和氨基糖苷类的活性。 在我们的第一个具体目标中，我们提出（a）推进这种噻唑烷酮化学型以定义一种新的噻唑烷酮化学型。 通过结合SAR研究和靶点分析用于治疗开发先导化合物 在体外基准研究和动物模型中的化合物;（B）表征结构机制 通过该方法噻唑烷酮抑制细菌分泌系统， 生物化学方法;以及（c）鉴定细菌分泌的另外的广谱抑制剂 通过高通量筛选和相应的二次测定的系统 在我们的第二个具体目标中，（a）作为c-di-GMP的潜在功能性抑制剂的化合物 将在生化试验中检测酶抑制和di-c-GMP（B）受体结合抑制 抑制c-di-GMP代谢和抑制多种c-di-GMP调节的性质将是 使用功能测定法测定化合物对活细菌的影响，以及（c）新化合物 将开发出通过高性能抑制di-c-GMP代谢成分的药物， 吞吐量和虚拟屏幕。 在我们的第三个具体目标中，我们提出（a）研究增加灵敏度的化合物， 涉及阳离子抗生素，用于使用动物模型抑制细菌感染的能力;（B）鉴定 通过重复我们的高通量筛选增加对抗生素敏感性的其他化学型 与Burkholderia thailandensis，一种对多粘菌素具有组成性抗性的模式非BSL 3生物， 其他抗生素的种类;及（c）确定相关化合物的作用机制 用遗传学方法分离耐药菌株提高对多粘菌素和氨基糖苷类药物敏感性 变种人

The antibiotic discovery project combines chemistry and biology to identify new compounds of utility in treating or preventing disease caused by Gram-negative bacteria. The project develops high throughput screens, performs chemical synthesis as part of hit to lead chemistry, measures the biological effect of compounds with purified enzymes and in whole bacteria, and studies compound efficacy in small animal models of disease. Currently three separate programs are active: (1) Gram-negative virulence proteins secretion inhibition by thiazolidinones (2) Diverse strategies including chemical synthesis of analogs and high throughput screen development to discover compounds which inhibit the three steps of the second messenger di-c-GMP metabolism and (3) Testing of a quinolone that has no intrinsic antibiotic activity but promotes the activity of the cationic antibiotics polymyxins and aminoglycosides. In our first specific aim we propose to (a) advance this thiazolidinone chemotype to define a lead compound for therapeutic development by using a combination of SAR studies and analysis of target compounds in in vitro benchmark studies and animal models; (b) characterize the structural mechanisms by which thiazolidinones inhibit bacterial secretion systems implementing a combination of genetic and biochemical approaches; and to (c) identify additional broad spectrum inhibitors of bacterial secretion systems by means of high-throughput screens and corresponding secondary assays In our second specific aim (a) compounds that are potential functional inhibitors of c-di-GMP will be tested in biochemical assays for enzyme inhibition and inhibition of receptor binding of di-c-GMP (b) inhibition of c-di-GMP metabolism and inhibition of a variety of c-di-GMP regulated properties will be determined using functional assays for the effect of compounds on live bacteria and (c) new compounds will be developed that inhibit the components of di-c-GMP metabolism by performance of high throughput and virtual screens. In our third specific aim we propose to (a) investigate compounds that that increase sensitivity to cationic antibiotics for the ability to inhibit bacterial infections using animal models; (b) identify additional chemotypes that increase sensitivity to antibiotics by repeating our high-throughput screen with Burkholderia thailandensis, a model non-BSL3 organism with constitutive resistance to polymyxin and a variety of other antibiotics; and (c) identify the mechanism of action by which relevant compounds increase sensitivity to polymyxin and aminoglycosides by using a genetic approach to isolate resistant mutants.