A High-Throughput Screening Assay for Fungicidal Compounds against Cryptococcus neoformans

作者：Jennifer L.A. Rabjohns, Yoon‐Dong Park, Jean Dehdashti, Wei Sun, Christina Henderson, Adrian M. Zelazny, Steven J. Metallo, Wei Zheng, Peter R. Williamson · 发表于：SLAS DISCOVERY · 年份：2013 · DOI：10.1177/1087057113496847 · 被引用次数：33 · 研究领域：Fungal Infections and Studies、Antifungal resistance and susceptibility、Nail Diseases and Treatments

Cryptococcus neoformans is a pathogenic fungus that causes meningitis worldwide, particularly in human immunodeficiency virus (HIV)-infected individuals. Although amphotericin B is the "gold standard" treatment for cryptococcal meningitis, the toxicity and inconvenience of intravenous injection emphasize a need for development of new anticryptocccal drugs. Recent data from humans and animal studies suggested that a nutrient-deprived host environment may exist in cryptococcal meningitis. Thus, a screening assay for identifying fungicidal compounds under nutrient-deprived conditions may provide an alternative strategy to develop new anticryptococcal drugs for this disease. A high-throughput fungicidal assay was developed using a profluorescent dye, alamarBlue, to detect residual metabolic activity of C. neoformans under nutrient-limiting conditions. Screening the Library of Pharmacologically Active Compounds (LOPAC) with this assay identified a potential chemical scaffold, 10058-F4, that exhibited fungicidal activity in the low micromolar range. These results thus demonstrate the feasibility of this alamarBlue-based assay for high-throughput screening of fungicidal compounds under nutrient-limiting conditions for new anticryptococcal drug development.