Novel Antibiofilm Treatments for Pseudomonas aeruginosa Infection

摘要/项目摘要 CDC报告说，2019年，美国发生了280万例抗生素耐药菌感染， 造成超过三万五千人死亡细菌生物膜是造成许多这些感染的原因， 通过非特异性因素，如药物的排除和改变，赋予对多种抗生素的耐药性 细菌代谢多重耐药铜绿假单胞菌是一种强生物膜形成病原体， 2019年有32，000人感染，2，700人死亡。这些感染大多与 铜绿假单胞菌显示出由复杂生物膜赋予的非特异性抗生素抗性 它形成了。为了应对这种病原体日益增加的威胁，必须采取创新的干预战略， 设计了这些策略之一是与目前使用的抗生素协同作用。我们采用高吞吐量 化学文库的筛选（HTS）和筛选复杂的平台产物以鉴定协同的 有效根除预形成的生物膜的化合物和物质。6000多步高温超导 合成化合物确定了两种有希望的候选物，三氯生（TRI）和羟氯扎胺（OXY）， 与抗生素妥布霉素（TOB）协同对抗已建立的铜绿假单胞菌生物膜。三氯生是一种 FDA批准的经过充分研究的抗菌剂然而，尚未研究其与TOB的协同作用。在 此外，植物源物质的筛选确定了Larrea tridentata和Hydratis的成分 加拿大假单胞菌能够根除预先形成的铜绿假单胞菌生物膜。小檗碱（BER），一种化合物 由Hydratis公司生产，在我们的试验中抑制铜绿假单胞菌生物膜，并且以前已经证明， 与TOB协同作用。我们建议在生物膜伤口感染模型中测试这种协同作用， 完了我们现在已经表明，Larrea的提取物，也被称为杂酚油（CRE）， 活性对建立铜绿假单胞菌生物膜，我们已经表明，这种提取物是有效的，在小鼠 铜绿假单胞菌感染的生物膜伤口模型。该模型基于体内生物发光成像 (BLI)这大大加快了动物体内生物膜感染的体内测试过程。生物发光P. 铜绿假单胞菌在伤口上方结痂的下侧形成生物膜，从而可以量化感染 通过非侵入性成像，在这里，我们建议开发治疗方法， 已经通过测试化合物和提取物在BLI伤口模型中的TOB协同作用来鉴定。这 该方法将以快速和定量的方式建立铜绿假单胞菌生物膜感染的新治疗方法。

Abstract/Project Summary The CDC reported that in 2019, 2.8 million infections due to antibiotic resistant bacteria occurred in the US, causing over 35,000 deaths. Bacterial biofilms are responsible for many of these infections, as biofilms confer resistance to multiple antibiotics via nonspecific factors such as exclusion of the drug and altered bacterial metabolism. Multidrug resistant Pseudomonas aeruginosa, a strong biofilm-forming pathogen, was responsible for 32,000 infections and 2,700 deaths in 2019. Most of these infections are associated with biofilms, and P. aeruginosa displays the nonspecific antibiotic resistance conferred by the complex biofilm it forms. To combat the increasing threat of this pathogen innovative intervention strategies must be designed. One of these strategies is synergy with currently used antibiotics. We employ high throughput screens (HTS) of chemical libraries and screening of complex plat products to identify synergistic compounds and substances that effectively eradicate preformed biofilms. A multistep HTS of over 6,000 synthetic compounds identified two promising candidates, triclosan (TRI) and oxyclozanide (OXY), that synergize with the antibiotic tobramycin (TOB) against established P. aeruginosa biofilms. Triclosan is a well-studied antimicrobial approved by the FDA. However it has not been studied for synergy with TOB. In addition, a screen of plant-derived substances identified Larrea tridentata and a component of Hydrastis canadensis as capable of eradicating preformed P. aeruginosa biofilms. Berberine (BER), a compound made by Hydrastis, inhibited P. aeruginosa biofilms in our assays and has previously been shown to synergize with TOB. We propose to test this synergy in a biofilm wound infection model, which has not been done. We have now shown that extracts of Larrea, which is also known as Creosote (CRE), are highly active against established P. aeruginosa biofilms, and we have shown this extract to be effective in a murine biofilm wound model of P. aeruginosa infection. This model is based on in vivo bioluminescence imaging (BLI), which greatly speeds the process of in vivo testing of biofilm infection in animals. Bioluminescent P. aeruginosa form a biofilm on the underside of a scab over the wound, allowing quantification of the infection in individual animals over time by non-invasive imaging. Here, we propose to develop the treatments we have identified by testing TOB synergy of the compounds and extracts in the BLI wound model. This process will establish novel treatments for P. aeruginosa biofilm infections in a rapid and quantitative manner.