Amino acid-derived 1,2-benzisothiazolinone derivatives as novel small-molecule antifungal inhibitors: identification of potential genetic targets.
We have identified four synthetic compounds (DFD-VI-15, BD-I-186, DFD-V-49, and DFD-V-66) from an amino acid-derived 1,2-benzisothiazolinone (BZT) scaffold that have reasonable MIC(50) values against a panel of fungal pathogens. These compounds have no structural similarity to existing antifungal drugs. Three of the four compounds have fungicidal activity against Candida spp., Cryptococcus neoformans, and several dermatophytes, while one is fungicidal to Aspergillus fumigatus. The kill rates of our compounds are equal to those in clinical usage. The BZT compounds remain active against azole-, polyene-, and micafungin-resistant strains of Candida spp. A genetics-based approach, along with phenotype analysis, was used to begin mode of action (MOA) studies of one of these compounds, DFD-VI-15. The genetics-based screen utilized a homozygous deletion collection of approximately 4,700 Saccharomyces cerevisiae mutants. We identified mutants that are both hypersensitive and resistant. Using FunSpec, the hypersensitive mutants and a resistant ace2 mutant clustered within a category of genes related directly or indirectly to mitochondrial functions. In Candida albicans, the functions of the Ace2p transcription factor include the regulation of glycolysis. Our model is that DFD-VI-15 targets a respiratory pathway that limits energy production. Supporting this hypothesis are phenotypic data indicating that DFD-VI-15 causes increased cell-reactive oxidants (ROS) and a decrease in mitochondrial membrane potential. Also, the same compound has activity when cells are grown in a medium containing glycerol (mitochondrial substrate) but is much less active when cells are grown anaerobically.
Georgetown University Medical Center, Washington, DC, USA.
Deepu
D
Alex
Francoise
F
Gay-Andrieu
Jared
J
May
Linta
L
Thampi
Dengfeng
D
Dou
Aileen
A
Mooney
William
W
Groutas
Richard
R
Calderone
D000596:Amino Acids / Q000138:chemical synthesis / Q000494:pharmacology*; D000935:Antifungal Agents / Q000138:chemical synthesis / Q000494:pharmacology*; D003883:Arthrodermataceae / Q000187:drug effects / Q000254:growth & development; D001232:Aspergillus fumigatus / Q000187:drug effects / Q000254:growth & development; D002176:Candida albicans / Q000187:drug effects / Q000254:growth & development; D003455:Cryptococcus neoformans / Q000187:drug effects / Q000254:growth & development; D026141:Drug Resistance, Multiple, Fungal / Q000187:drug effects; D005656:Fungal Proteins / Q000235:genetics* / Q000378:metabolism; D005990:Glycerol / Q000378:metabolism; D006019:Glycolysis / Q000187:drug effects / Q000235:genetics; D053078:Membrane Potential, Mitochondrial / Q000187:drug effects; D008826:Microbial Sensitivity Tests; D008928:Mitochondria / Q000187:drug effects / Q000235:genetics / Q000378:metabolism; D009154:Mutation; D012441:Saccharomyces cerevisiae / Q000187:drug effects / Q000235:genetics* / Q000378:metabolism; D013844:Thiazoles / Q000138:chemical synthesis / Q000494:pharmacology*; D014157:Transcription Factors / Q000235:genetics* / Q000378:metabolism
D000596:Amino Acids; D000935:Antifungal Agents; D005656:Fungal Proteins; D013844:Thiazoles; D014157:Transcription Factors; C015699:1,2-benzisothiazoline-3-one; D005990:Glycerol