title: Identification of a New Class of Antifungals Targeting the Synthesis of Fungal Sphingolipids.
abstract: Recent estimates suggest that >300 million people are afflicted by serious fungal infections worldwide. Current antifungal drugs are static and toxic and/or have a narrow spectrum of activity. Thus, there is an urgent need for the development of new antifungal drugs. The fungal sphingolipid glucosylceramide (GlcCer) is critical in promoting virulence of a variety of human-pathogenic fungi. In this study, we screened a synthetic drug library for compounds that target the synthesis of fungal, but not mammalian, GlcCer and found two compounds [N'-(3-bromo-4-hydroxybenzylidene)-2-methylbenzohydrazide (BHBM) and its derivative, 3-bromo-N'-(3-bromo-4-hydroxybenzylidene) benzohydrazide (D0)] that were highly effective in vitro and in vivo against several pathogenic fungi. BHBM and D0 were well tolerated in animals and are highly synergistic or additive to current antifungals. BHBM and D0 significantly affected fungal cell morphology and resulted in the accumulation of intracellular vesicles. Deep-sequencing analysis of drug-resistant mutants revealed that four protein products, encoded by genes APL5, COS111, MKK1, and STE2, which are involved in vesicular transport and cell cycle progression, are targeted by BHBM.
  
  
  OBJECTIVE
  Fungal infections are a significant cause of morbidity and mortality worldwide. Current antifungal drugs suffer from various drawbacks, including toxicity, drug resistance, and narrow spectrum of activity. In this study, we have demonstrated that pharmaceutical inhibition of fungal glucosylceramide presents a new opportunity to treat cryptococcosis and various other fungal infections. In addition to being effective against pathogenic fungi, the compounds discovered in this study were well tolerated by animals and additive to current antifungals. These findings suggest that these drugs might pave the way for the development of a new class of antifungals.
authors.affiliation: Department of Molecular Genetics and Microbiology, Stony Brook University, Stony Brook, New York, USA.
authors.forename: Visesato
authors.initials: V
authors.lastname: Mor
authors.affiliation: Department of Molecular Genetics and Microbiology, Stony Brook University, Stony Brook, New York, USA.
authors.forename: Antonella
authors.initials: A
authors.lastname: Rella
authors.affiliation: Department of Molecular Genetics and Microbiology, Stony Brook University, Stony Brook, New York, USA.
authors.forename: Amir M
authors.initials: AM
authors.lastname: Farnoud
authors.affiliation: Department of Molecular Genetics and Microbiology, Stony Brook University, Stony Brook, New York, USA.
authors.forename: Ashutosh
authors.initials: A
authors.lastname: Singh
authors.affiliation: Department of Molecular Genetics and Microbiology, Stony Brook University, Stony Brook, New York, USA.
authors.forename: Mansa
authors.initials: M
authors.lastname: Munshi
authors.affiliation: Department of Molecular Genetics and Microbiology, Stony Brook University, Stony Brook, New York, USA.
authors.forename: Arielle
authors.initials: A
authors.lastname: Bryan
authors.affiliation: Department of Molecular Genetics and Microbiology, Stony Brook University, Stony Brook, New York, USA.
authors.forename: Shamoon
authors.initials: S
authors.lastname: Naseem
authors.affiliation: Department of Molecular Genetics and Microbiology, Stony Brook University, Stony Brook, New York, USA.
authors.forename: James B
authors.initials: JB
authors.lastname: Konopka
authors.affiliation: Department of Chemistry and Institute of Chemical Biology and Drug Discovery, Stony Brook University, Stony Brook, New York, USA.
authors.forename: Iwao
authors.initials: I
authors.lastname: Ojima
authors.affiliation: Department of Biochemistry and Molecular Biology, Medical University of South Carolina, Charleston, South Carolina, USA.
authors.forename: Erika
authors.initials: E
authors.lastname: Bullesbach
authors.affiliation: Department of Veterans Affairs Medical Center, Cincinnati, Ohio, USA.
authors.forename: Alan
authors.initials: A
authors.lastname: Ashbaugh
authors.forename: Michael J
authors.initials: MJ
authors.lastname: Linke
authors.forename: Melanie
authors.initials: M
authors.lastname: Cushion
authors.affiliation: University of Cincinnati College of Medicine, Cincinnati, Ohio, USA.
authors.forename: Margaret
authors.initials: M
authors.lastname: Collins
authors.affiliation: Department of Pharmaceutical Sciences, University of Cincinnati, Cincinnati, Ohio, USA.
authors.forename: Hari Krishna
authors.initials: HK
authors.lastname: Ananthula
authors.affiliation: Department of Chemistry, University of Cincinnati, Cincinnati, Ohio, USA.
authors.forename: Larry
authors.initials: L
authors.lastname: Sallans
authors.affiliation: Department of Pharmaceutical Sciences, University of Cincinnati, Cincinnati, Ohio, USA.
authors.forename: Pankaj B
authors.initials: PB
authors.lastname: Desai
authors.affiliation: Department of Pathology, Fungus Testing Laboratory, University of Texas Health Science Center at San Antonio, San Antonio, Texas, USA.
authors.forename: Nathan P
authors.initials: NP
authors.lastname: Wiederhold
authors.affiliation: Department of Pathology, Fungus Testing Laboratory, University of Texas Health Science Center at San Antonio, San Antonio, Texas, USA.
authors.forename: Annette W
authors.initials: AW
authors.lastname: Fothergill
authors.affiliation: Division of Infectious Diseases, University of Texas Health Science Center at San Antonio, San Antonio, Texas, USA.
authors.forename: William R
authors.initials: WR
authors.lastname: Kirkpatrick
authors.affiliation: Division of Infectious Diseases, University of Texas Health Science Center at San Antonio, San Antonio, Texas, USA.
authors.forename: Thomas
authors.initials: T
authors.lastname: Patterson
authors.affiliation: Department of Pharmaceutical Sciences, University of British Columbia, Vancouver, British Colombia, Canada.
authors.forename: Lai Hong
authors.initials: LH
authors.lastname: Wong
authors.affiliation: Department of Pharmaceutical Sciences, University of British Columbia, Vancouver, British Colombia, Canada.
authors.forename: Sunita
authors.initials: S
authors.lastname: Sinha
authors.affiliation: Department of Pharmaceutical Sciences, University of British Columbia, Vancouver, British Colombia, Canada.
authors.forename: Guri
authors.initials: G
authors.lastname: Giaever
authors.affiliation: Department of Pharmaceutical Sciences, University of British Columbia, Vancouver, British Colombia, Canada.
authors.forename: Corey
authors.initials: C
authors.lastname: Nislow
authors.affiliation: Department of Biomedical Engineering, Worcester Polytechnic Institute, Worcester, Massachusetts, USA.
authors.forename: Patrick
authors.initials: P
authors.lastname: Flaherty
authors.affiliation: Department of Biochemistry and Molecular Biology, Baylor College of Medicine, Houston, Texas, USA.
authors.forename: Xuewen
authors.initials: X
authors.lastname: Pan
authors.affiliation: Instituto de Microbiologia Professor Paulo de Góes, Universidade Federal do Rio de Janeiro, Rio de Janeiro, Brazil.
authors.forename: Gabriele Vargas
authors.initials: GV
authors.lastname: Cesar
authors.affiliation: Instituto de Microbiologia Professor Paulo de Góes, Universidade Federal do Rio de Janeiro, Rio de Janeiro, Brazil.
authors.forename: Patricia
authors.initials: P
authors.lastname: de Melo Tavares
authors.affiliation: Laboratório de Ultraestrutura Celular Hertha Meyer, Instituto de Biofísica Carlos Chagas Filho, Universidade Federal do Rio de Janeiro, Rio de Janeiro, Brazil.
authors.forename: Susana
authors.initials: S
authors.lastname: Frases
authors.forename: Kildare
authors.initials: K
authors.lastname: Miranda
authors.forename: Marcio L
authors.initials: ML
authors.lastname: Rodrigues
authors.affiliation: Department of Physiology and Biophysics, Stony Brook University, Stony Brook, New York, USA.
authors.forename: Chiara
authors.initials: C
authors.lastname: Luberto
authors.affiliation: Instituto de Microbiologia Professor Paulo de Góes, Universidade Federal do Rio de Janeiro, Rio de Janeiro, Brazil.
authors.forename: Leonardo
authors.initials: L
authors.lastname: Nimrichter
authors.affiliation: Department of Molecular Genetics and Microbiology, Stony Brook University, Stony Brook, New York, USA maurizio.delpoeta@stonybrook.edu.
authors.forename: Maurizio
authors.initials: M
authors.lastname: Del Poeta
mesh_terms: D000818:Animals; D000935:Antifungal Agents / Q000009:adverse effects / Q000302:isolation & purification* / Q000494:pharmacology* / Q000633:toxicity; D001593:Benzyl Compounds / Q000009:adverse effects / Q000302:isolation & purification* / Q000494:pharmacology* / Q000633:toxicity; D053898:Biosynthetic Pathways / Q000187:drug effects*; D002177:Candidiasis / Q000188:drug therapy; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D015169:Colony Count, Microbial; D004195:Disease Models, Animal; D004353:Drug Evaluation, Preclinical / Q000379:methods; D004357:Drug Synergism; D064420:Drug-Related Side Effects and Adverse Reactions; D005658:Fungi / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism / Q000502:physiology; D008264:Macrophages / Q000187:drug effects / Q000502:physiology; D051379:Mice; D008826:Microbial Sensitivity Tests; D050296:Microbial Viability / Q000187:drug effects; D046529:Microscopy, Electron, Transmission; D015394:Molecular Structure; D013107:Sphingolipids / Q000037:antagonists & inhibitors / Q000096:biosynthesis*; D016896:Treatment Outcome
chemical_list: D000935:Antifungal Agents; D001593:Benzyl Compounds; D013107:Sphingolipids