title: 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.affiliation: Laboratory of Clinical Infectious Diseases, National Institutes of Health, Bethesda, MD, USA.
authors.forename: Jennifer L A
authors.initials: JLA
authors.lastname: Rabjohns
authors.affiliation: Laboratory of Clinical Infectious Diseases, National Institutes of Health, Bethesda, MD, USA.
authors.forename: Yoon-Dong
authors.initials: YD
authors.lastname: Park
authors.affiliation: National Center for Advancing Translational Sciences, National Institutes of Health, Bethesda, MD, USA.
authors.forename: Jean
authors.initials: J
authors.lastname: Dehdashti
authors.affiliation: Department of Laboratory Medicine, Clinical Center, National Institutes of Health, Bethesda, MD USA.
authors.forename: Christina
authors.initials: C
authors.lastname: Henderson
authors.affiliation: Department of Laboratory Medicine, Clinical Center, National Institutes of Health, Bethesda, MD USA.
authors.forename: Adrian
authors.initials: A
authors.lastname: Zelazny
authors.affiliation: Chemistry Department, Georgetown University, Washington D.C., USA.
authors.forename: Steven J
authors.initials: SJ
authors.lastname: Metallo
authors.affiliation: National Center for Advancing Translational Sciences, National Institutes of Health, Bethesda, MD, USA.
authors.forename: Wei
authors.initials: W
authors.lastname: Zheng
authors.affiliation: Laboratory of Clinical Infectious Diseases, National Institutes of Health, Bethesda, MD, USA.|Section of Infectious Diseases, Department of Medicine, University of Illinois at Chicago, IL, USA.
authors.forename: Peter R
authors.initials: PR
authors.lastname: Williamson
mesh_terms: D000666:Amphotericin B / Q000627:therapeutic use; D000935:Antifungal Agents / Q000737:chemistry / Q000302:isolation & purification* / Q000627:therapeutic use; D003455:Cryptococcus neoformans / Q000187:drug effects* / Q000472:pathogenicity; D057166:High-Throughput Screening Assays; D006801:Humans; D016919:Meningitis, Cryptococcal / Q000188:drug therapy / Q000382:microbiology; D054852:Small Molecule Libraries; D013844:Thiazoles / Q000737:chemistry / Q000302:isolation & purification* / Q000627:therapeutic use
chemical_list: C524814:5-(4-ethylbenzylidene)-2-thioxothiazolidin-4-one; D000935:Antifungal Agents; D054852:Small Molecule Libraries; D013844:Thiazoles; D000666:Amphotericin B
keywords: Cryptococcus neoformans; alamarBlue assay; fungicidal screen; high-throughput screen
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