以白色念珠菌为模型高通量筛选潜在抗真菌药物

 目的通过高通量药物筛选来寻找有活性的广谱性抗真菌药物,专一性抗白色念珠菌药物以及特异性抑制含核酶的白色念珠菌生长的药物。方法以两株基因型不同的白色念珠菌菌株(一株含I型内含子核酶,一株不含核酶)和一株非病原真菌啤酒酵母(不含I型内含子核酶)为细胞模型,3种菌株依次以96孔板为容器,在起始菌液浓度的A_580nm为0.1时分别加入10720种药物样品(包含化合物与天然产物),在30℃静止培养6h,选取抑菌效率超过50%的样品进行复筛。结果通过初筛和复筛,发现了23种药物在10μg·mL^-1的浓度时的抑菌效率超过50%,其中1种药物特异性抑制含核酶的白色念珠菌生长,是潜在的抗核酶药物;4种药物特异抑制白色念珠菌的生长,是潜在的抗白色念珠菌药物;18种药物抑制所有模式菌株的生长,是潜在的广谱性抗真菌药物。结论本研究在国内首次采用自动化工作站进行抗真菌药物的高通量药物筛选,方法简便快捷,筛选结果稳定可靠,并筛选到一些有希望向临床发展的抗真菌药物活性样品。此方法有可能应用于抗其他病原微生物的药物筛选。

High-throughput screen of potential antifungal drugs using Candida albicans as a model

OBJECTIVE To discover broad-spectrum antifungal drugs,specific anti-Candida albicnas drugs and specific inhibitors of the group Ⅰ intron-containing Candida albicans,by means of the model consisting of three different yeast strains (one is the intron-containing Candida albicans,one is the intronless Candida albicans,one is the intronless Saccharomyces cerevisiae).METHODS 10 720 Samples obtained from the national drug library of China,including natural products and synthetic compounds, were tested with the three strains for their inhibitory efficiency respectively. After static incubation of the yeast in the presence of drug samples at 30℃ for 6 hours,the samples whose inhibitory efficiency was over 50% were selected for secondary screening.RESULTS After primary and secondary screening, the inhibitory efficiency of 23 active samples was over 50% at 10 μg·mL^-1,one of these active samples specifically inhibited the growth of intron-containing Candida albicans,four specifically inhibited the growth of Candida albicans, eighteen inhibited the growth of all yeasts.CONCLUSION This study represented the first high-throughput screening method to antifungal drug discovery in China.The screening of national drug library inChina firstly allow us to identify several new antifungal compounds that potentially was developed for clinical use. Because of its simplicity and stable results, this method can give implication for the drug discovery of other pathogenic strains.