Kumar Anand, Drozd Mary, Pina-Mimbela Ruby, Xu Xiulan, Helmy Yosra A, Antwi Janet, Fuchs James R, Nislow Corey, Templeton Jillian, Blackall Patrick J, Rajashekara Gireesh
Food Animal Health Research Program, Department of Veterinary Preventive Medicine, The Ohio State UniversityWooster, OH, USA; Poultry CRC, University of New EnglandArmidale, NSW, Australia.
Food Animal Health Research Program, Department of Veterinary Preventive Medicine, The Ohio State University Wooster, OH, USA.
Front Microbiol. 2016 Apr 6;7:405. doi: 10.3389/fmicb.2016.00405. eCollection 2016.
Campylobacter is a leading cause of foodborne bacterial gastroenteritis worldwide and infections can be fatal. The emergence of antibiotic-resistant Campylobacter spp. necessitates the development of new antimicrobials. We identified novel anti-Campylobacter small molecule inhibitors using a high throughput growth inhibition assay. To expedite screening, we made use of a "bioactive" library of 4182 compounds that we have previously shown to be active against diverse microbes. Screening for growth inhibition of Campylobacter jejuni, identified 781 compounds that were either bactericidal or bacteriostatic at a concentration of 200 μM. Seventy nine of the bactericidal compounds were prioritized for secondary screening based on their physico-chemical properties. Based on the minimum inhibitory concentration against a diverse range of C. jejuni and a lack of effect on gut microbes, we selected 12 compounds. No resistance was observed to any of these 12 lead compounds when C. jejuni was cultured with lethal or sub-lethal concentrations suggesting that C. jejuni is less likely to develop resistance to these compounds. Top 12 compounds also possessed low cytotoxicity to human intestinal epithelial cells (Caco-2 cells) and no hemolytic activity against sheep red blood cells. Next, these 12 compounds were evaluated for ability to clear C. jejuni in vitro. A total of 10 compounds had an anti-C. jejuni effect in Caco-2 cells with some effective even at 25 μM concentrations. These novel 12 compounds belong to five established antimicrobial chemical classes; piperazines, aryl amines, piperidines, sulfonamide, and pyridazinone. Exploitation of analogs of these chemical classes may provide Campylobacter specific drugs that can be applied in both human and animal medicine.
弯曲杆菌是全球食源性细菌性肠胃炎的主要病因,感染可能致命。抗生素耐药性弯曲杆菌属的出现使得开发新型抗菌药物成为必要。我们使用高通量生长抑制试验鉴定了新型抗弯曲杆菌小分子抑制剂。为了加快筛选速度,我们利用了一个由4182种化合物组成的“生物活性”文库,我们之前已证明该文库对多种微生物具有活性。对空肠弯曲杆菌的生长抑制进行筛选,确定了781种在200μM浓度下具有杀菌或抑菌作用的化合物。根据其物理化学性质,对79种杀菌化合物进行了二次筛选。基于对多种空肠弯曲杆菌的最低抑菌浓度以及对肠道微生物无影响,我们选择了12种化合物。当空肠弯曲杆菌与致死或亚致死浓度培养时,未观察到对这12种先导化合物产生耐药性,这表明空肠弯曲杆菌对这些化合物产生耐药性的可能性较小。排名前12的化合物对人肠上皮细胞(Caco-2细胞)也具有低细胞毒性,并且对绵羊红细胞无溶血活性。接下来,评估这12种化合物在体外清除空肠弯曲杆菌的能力。共有10种化合物在Caco-2细胞中具有抗空肠弯曲杆菌的作用,有些化合物甚至在25μM浓度下仍有效。这12种新型化合物属于五个已确立的抗菌化学类别;哌嗪类、芳基胺类、哌啶类、磺胺类和哒嗪酮类。对这些化学类别的类似物进行开发可能会提供可应用于人类和兽医学的针对弯曲杆菌的特异性药物。