Antibiotic Drug Discovery

抗生素发现项目将化学和生物学结合起来，以确定在 治疗或预防由革兰氏阴性细菌引起的疾病。该项目开发了高生产能力 筛选、执行化学合成作为HIT to Lead化学的一部分，测量 含有纯化酶的化合物和整个细菌中的化合物，并研究化合物在小动物中的效果 疾病的模型。目前有三个单独的程序是有效的：(1)革兰氏阴性毒力蛋白 噻唑烷酮类化合物的分泌抑制作用(2)包括化学合成类似物在内的多种策略 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)一种没有内在抗生素的喹诺酮类药物的检测 活性，但可促进阳离子抗生素、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，我们建议(A)推进这种噻唑烷酮化学类型以定义一个 利用合成孔径雷达研究和靶标分析相结合的方法开发治疗用先导化合物 体外基准研究和动物模型中的化合物；(B)表征结构机制 通过这种方式，噻唑烷酮类化合物抑制细菌分泌系统，实现遗传和 生物化学方法；以及(C)确定更多的广谱细菌分泌抑制剂 通过高通量筛选和相应的二次化验建立系统 在我们的第二个特定目标(A)中，化合物是c-di-GMP的潜在功能抑制剂 将在生化分析中进行酶抑制和对di-c-GMP受体结合的抑制试验(B) C-di-GMP代谢的抑制和c-di-GMP的多种调节特性的抑制 使用功能分析测定化合物对活细菌的影响和(C)新化合物 将开发通过高性能抑制di-c-GMP代谢的成分 吞吐量和虚拟屏幕。 在我们第三个具体目标中，我们建议：(A)研究那些增加敏感性的化合物 利用动物模型研究阳离子抗生素抑制细菌感染的能力；(B)确定 通过重复我们的高通量筛查来增加对抗生素敏感性的其他化学类型 泰兰伯克霍尔德氏菌是一种对多粘菌素具有结构性抗性的非BSL3模式生物 其他抗生素的种类；以及(C)确定相关化合物的作用机制 用遗传方法分离耐药株提高对多粘菌素和氨基糖苷类药物的敏感性 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.