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

Therapeutic Approaches

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

Rabjohns, Jennifer L A; Park, Yoon-Dong; Dehdashti, Jean; Sun, Wei; Henderson, Christina; Zelazny, Adrian; Metallo, Steven J; Zheng, Wei; Williamson, Peter R;

Keywords

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

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