title: High-Throughput Screen inCryptococcus neoformansIdentifies a Novel Molecular Scaffold That Inhibits Cell Wall Integrity Pathway Signaling.
abstract: Cryptococcus neoformansis one of the most important human fungal pathogens; however, no new therapies have been developed in over 50 years. Fungicidal activity is crucially important for an effective anticryptococal agent and, therefore, we screened 361,675 molecules againstC. neoformansusing an adenylate kinase release assay that specifically detects fungicidal activity. A set of secondary assays narrowed the set of hits to molecules that interfere with fungal cell wall integrity and identified three benzothioureas with low in vitro mammalian toxicity and good in vitro anticryptococcal (minimum inhibitory concentration = 4 μg/mL). This scaffold inhibits signaling through the cell wall integrity MAP kinase cascade. Structure-activity studies indicate that the thiocarbonyl moiety is crucial for activity. Genetic and biochemical data suggest that benzothioureas inhibit signaling upstream of the kinase cascade. Thus, the benzothioureas appear to be a promising new scaffold for further exploration in the search for new anticryptococcal agents.
authors.affiliation: Center for the Science of Therapeutics, Broad Institute of Harvard and MIT , 7 Cambridge Center, Cambridge, Massachusetts 02142, United States.
authors.forename: Kate
authors.initials: K
authors.lastname: Hartland
authors.affiliation: Center for the Science of Therapeutics, Broad Institute of Harvard and MIT , 7 Cambridge Center, Cambridge, Massachusetts 02142, United States.
authors.forename: Jun
authors.initials: J
authors.lastname: Pu
authors.affiliation: Center for the Science of Therapeutics, Broad Institute of Harvard and MIT , 7 Cambridge Center, Cambridge, Massachusetts 02142, United States.
authors.forename: Michelle
authors.initials: M
authors.lastname: Palmer
authors.affiliation: Center for the Science of Therapeutics, Broad Institute of Harvard and MIT , 7 Cambridge Center, Cambridge, Massachusetts 02142, United States.
authors.forename: Sivaraman
authors.initials: S
authors.lastname: Dandapani
authors.affiliation: Center for the Science of Therapeutics, Broad Institute of Harvard and MIT , 7 Cambridge Center, Cambridge, Massachusetts 02142, United States.
authors.forename: Philip N
authors.initials: PN
authors.lastname: Moquist
authors.affiliation: Center for the Science of Therapeutics, Broad Institute of Harvard and MIT , 7 Cambridge Center, Cambridge, Massachusetts 02142, United States.
authors.forename: Benito
authors.initials: B
authors.lastname: Munoz
authors.affiliation: Department of Pediatrics and Department of Microbiology/Immunology, University of Rochester School of Medicine and Dentistry , 601 Elmwood Avenue, Rochester, New York 14642, United States.
authors.forename: Louis
authors.initials: L
authors.lastname: DiDone
authors.affiliation: Center for the Science of Therapeutics, Broad Institute of Harvard and MIT , 7 Cambridge Center, Cambridge, Massachusetts 02142, United States.
authors.forename: Stuart L
authors.initials: SL
authors.lastname: Schreiber
authors.affiliation: Department of Pediatrics and Department of Microbiology/Immunology, University of Rochester School of Medicine and Dentistry, 601 Elmwood Avenue, Rochester, New York 14642, United States; Department of Pediatrics and Department of Microbiology/Immunology, University of Rochester School of Medicine and Dentistry, 601 Elmwood Avenue, Rochester, New York 14642, United States.
authors.forename: Damian J
authors.initials: DJ
authors.lastname: Krysan
keywords: Cryptococcus neoformans; antifungal; cell wall integrity pathway; high-throughput screening; mitogen-activated protein kinase; yeast cell wall