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J Biomol Screen. 2014 Feb;19(2):270-277. doi: 10.1177/1087057113496847. Epub 2013 Jul 29.

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

Author information

1
Laboratory of Clinical Infectious Diseases, National Institutes of Health, Bethesda, MD, USA.
2
National Center for Advancing Translational Sciences, National Institutes of Health, Bethesda, MD, USA.
3
Department of Laboratory Medicine, Clinical Center, National Institutes of Health, Bethesda, MD USA.
4
Chemistry Department, Georgetown University, Washington D.C., USA.
5
Section of Infectious Diseases, Department of Medicine, University of Illinois at Chicago, IL, USA.
#
Contributed equally

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.

KEYWORDS:

Cryptococcus neoformans; alamarBlue assay; fungicidal screen; high-throughput screen

PMID:
23896686
PMCID:
PMC4017337
DOI:
10.1177/1087057113496847
[Indexed for MEDLINE]
Free PMC Article

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