CYTOMETRY-AIDED DEVELOPMENT OF ANTIMICROBIAL AGENTS

描述(申请人提供)：最近发生了一起令人震惊的事件 由于细菌对所有可用的抗生素产生抗药性而导致的感染增加。 四环素耐药性通常由外排泵介导；四环素 抑制它们的衍生品可以与现有的 四环素类药物用于治疗耐药生物的感染。流式细胞仪 将开发便于测量四环素摄入量的方法和 细菌外排，以及四环素和达托霉素对其影响的检测 膜电位、pH和通透性。另一种增强能力的方法 现有的抗生素是通过发现处理过的细菌 一些亚致死剂量的抗生素对以下染料具有瞬时渗透性 通常不能穿透细菌或哺乳动物的细胞膜。 用于使染料变得不耐用的化学改性可应用于 各种毒剂；这些修饰剂对宿主细胞应该是无毒的， 而是杀死瞬间渗透的细菌。这一假设可能会提供 针对细菌感染的一种新的通用治疗方法将是 使用荧光核酸结合化合物进行测试，已知的含义和 不必要的字符；对抗生素敏感和耐药的葡萄球菌和 将检查肠球菌和大肠埃希菌。 建议的商业应用： 制药公司和研究机构的分析和研究机构有很大的市场 促进新抗菌剂快速开发的相关仪器。 此外，改善感染治疗的药物或组合甚至 少数几种具有抗生素耐药性的细菌将捕获巨大的 抗菌剂市场。

DESCRIPTION (provided by applicant): There has recently been an alarming increase in infections due to bacteria resistant to all available antibiotics. Tetracycline resistance is typically mediated by efflux pumps; tetracycline derivatives which inhibit them may be used in combination with existing tetracyclines to treat infections by resistant organisms. Flow cytometric methods will be developed to facilitate measurement of tetracycline uptake and efflux in bacteria, and detection of effects of tetracyclines and daptomycin on membrane potential, pH, and permeability. Another method of potentiating existing antibiotics is suggested by the finding that bacteria treated with sublethal doses of some antibiotics become transiently permeable to dyes that normally do not penetrate the membranes of either bacterial or mammalian cells. The chemical modifications used to make dyes impermeant can be applied to various toxic agents; these modified agents should be nontoxic to host cells, but kill transiently permeabilized bacteria. This hypothesis, which may provide a novel and general therapeutic approach to bacterial infections, will be tested using fluorescent nucleic acid binding compounds with known permeant and impermeant characters; antibiotic-sensitive and -resistant Staphylococcus and Enterococcus species and Escherichia coli will be examined. PROPOSED COMMERCIAL APPLICATION: There is a substantial market in drug companies and research institutions for assays and related instrumentation which facilitate rapid development of new antimicrobial agents. Moreover, drugs or combinations which improved treatment of infections due to even a few antibiotic-resistant bacterial species would capture a significant share of a huge market for antibacterial agents.