CYTOMETRY-AIDED DEVELOPMENT OF ANTIMICROBIAL AGENTS

描述（由申请人提供）：最近有一个令人震惊的 由于细菌对所有可用的抗生素产生耐药性，感染增加。 四环素耐药通常由外排泵介导;四环素 抑制它们的衍生物可以与现有的 四环素类来治疗耐药生物体的感染。流式细胞术 将开发便于测量四环素摄取的方法， 细菌外排，以及检测四环素类和达托霉素对 膜电位、pH和渗透性。另一种增强 现有的抗生素是由发现， 亚致死剂量的一些抗生素变得对染料短暂可渗透， 通常不穿透细菌或哺乳动物细胞的膜。 用于使染料不渗透的化学改性可应用于 各种毒性试剂;这些修饰的试剂应该对宿主细胞无毒， 但是杀死瞬时透化的细菌。这一假设可能提供了 一种新的和一般的治疗方法，细菌感染，将 使用荧光核酸结合化合物与已知的渗透剂和 不渗透性特征;对抗生素敏感和耐药的葡萄球菌， 将检查肠球菌属和大肠埃希菌。 拟定商业应用： 在制药公司和研究机构中存在用于测定和分析的大量市场。 相关的仪器，促进新的抗微生物剂的快速发展。 此外，改善甚至由于感染而引起的感染的治疗的药物或组合物， 一些具有抗药性的细菌物种将占据一个巨大的 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.