A high-throughput screening assay for fungicidal compounds against Cryptococcus neoformans.

Abstract

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.

• Authors: Jennifer L A Rabjohns, Yoon-Dong Park, Jean Dehdashti, Christina Henderson, Adrian Zelazny, Steven J Metallo, Wei Zheng, Peter R Williamson
• Journal: Journal of biomolecular screening (J Biomol Screen) Vol. 19 Issue 2 Pg. 270-277 (Feb 2014) ISSN: 1552-454X [Electronic] United States
• PMID: 23896686 (Publication Type: Journal Article, Research Support, N.I.H., Intramural)

Chemical References

• 5-(4-ethylbenzylidene)-2-thioxothiazolidin-4-one
• Antifungal Agents
• Small Molecule Libraries
• Thiazoles
• Amphotericin B

Topics

• Amphotericin B (therapeutic use)
• Antifungal Agents (chemistry, isolation & purification, therapeutic use)
• Cryptococcus neoformans (drug effects, pathogenicity)
• High-Throughput Screening Assays
• Humans
• Meningitis, Cryptococcal (drug therapy, microbiology)
• Small Molecule Libraries
• Thiazoles (chemistry, isolation & purification, therapeutic use)