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Identification of antifungal compounds active against Candida albicans using an improved high-throughput Caenorhabditis elegans assay.

Okoli I, Coleman JJ, Tampakakis E, Tempakakis E, An WF, Holson E, Wagner F, Conery AL, Larkins-Ford J, Wu G, Stern A, Ausubel FM, Mylonakis E - PLoS ONE (2009)

Bottom Line: This high-throughput method was utilized to screen a library of 3,228 compounds represented by 1,948 bioactive compounds and 1,280 small molecules derived via diversity-oriented synthesis.Nineteen compounds were identified that conferred an increase in C. elegans survival, including most known antifungal compounds within the chemical library.In addition to seven clinically used antifungal compounds, twelve compounds were identified which are not primarily used as antifungal agents, including three immunosuppressive drugs.

View Article: PubMed Central - PubMed

Affiliation: Division of Infectious Diseases, Massachusetts General Hospital, Boston, Massachusetts, United States of America.

ABSTRACT
Candida albicans, the most common human pathogenic fungus, can establish a persistent lethal infection in the intestine of the microscopic nematode Caenorhabditis elegans. The C. elegans-C. albicans infection model was previously adapted to screen for antifungal compounds. Modifications to this screen have been made to facilitate a high-throughput assay including co-inoculation of nematodes with C. albicans and instrumentation allowing precise dispensing of worms into assay wells, eliminating two labor-intensive steps. This high-throughput method was utilized to screen a library of 3,228 compounds represented by 1,948 bioactive compounds and 1,280 small molecules derived via diversity-oriented synthesis. Nineteen compounds were identified that conferred an increase in C. elegans survival, including most known antifungal compounds within the chemical library. In addition to seven clinically used antifungal compounds, twelve compounds were identified which are not primarily used as antifungal agents, including three immunosuppressive drugs. This assay also allowed the assessment of the relative minimal inhibitory concentration, the effective concentration in vivo, and the toxicity of the compound in a single assay.

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Related in: MedlinePlus

Flow chart representing the C. elegans “curing” assay that identified 19 effective compounds.
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pone-0007025-g003: Flow chart representing the C. elegans “curing” assay that identified 19 effective compounds.

Mentions: After establishing the robustness and repeatability of the assay, a screen of 3,228 compounds was conducted representing compounds not screened in previous studies. This study, in combination with the previous screen by Breger et al (2007), brings the total number of screened compounds to 4,494. This collection was represented by 1,948 bioactive compounds consisting of FDA approved drugs and other compounds from the Microscorce Spectrum collection (Microsource Discovery Systems, Inc.) and the Prestwick compound library (Prestwick Chemicals, Inc.) and 1,280 small molecules derived via diversity-oriented synthesis [20], [21] from the Broad Institute of Harvard and MIT (Cambridge, MA). As shown in Table 1, the screening library represented a diverse collection of bioactive compounds composed of multiple chemical classes containing a broad array of functionality. In total, 19 compounds were identified in the screen (Figure 3). More importantly, the C. elegans screen was able to detect most of the well established antifungal agents (Table 1 and Table 2).


Identification of antifungal compounds active against Candida albicans using an improved high-throughput Caenorhabditis elegans assay.

Okoli I, Coleman JJ, Tampakakis E, Tempakakis E, An WF, Holson E, Wagner F, Conery AL, Larkins-Ford J, Wu G, Stern A, Ausubel FM, Mylonakis E - PLoS ONE (2009)

Flow chart representing the C. elegans “curing” assay that identified 19 effective compounds.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2737148&req=5

pone-0007025-g003: Flow chart representing the C. elegans “curing” assay that identified 19 effective compounds.
Mentions: After establishing the robustness and repeatability of the assay, a screen of 3,228 compounds was conducted representing compounds not screened in previous studies. This study, in combination with the previous screen by Breger et al (2007), brings the total number of screened compounds to 4,494. This collection was represented by 1,948 bioactive compounds consisting of FDA approved drugs and other compounds from the Microscorce Spectrum collection (Microsource Discovery Systems, Inc.) and the Prestwick compound library (Prestwick Chemicals, Inc.) and 1,280 small molecules derived via diversity-oriented synthesis [20], [21] from the Broad Institute of Harvard and MIT (Cambridge, MA). As shown in Table 1, the screening library represented a diverse collection of bioactive compounds composed of multiple chemical classes containing a broad array of functionality. In total, 19 compounds were identified in the screen (Figure 3). More importantly, the C. elegans screen was able to detect most of the well established antifungal agents (Table 1 and Table 2).

Bottom Line: This high-throughput method was utilized to screen a library of 3,228 compounds represented by 1,948 bioactive compounds and 1,280 small molecules derived via diversity-oriented synthesis.Nineteen compounds were identified that conferred an increase in C. elegans survival, including most known antifungal compounds within the chemical library.In addition to seven clinically used antifungal compounds, twelve compounds were identified which are not primarily used as antifungal agents, including three immunosuppressive drugs.

View Article: PubMed Central - PubMed

Affiliation: Division of Infectious Diseases, Massachusetts General Hospital, Boston, Massachusetts, United States of America.

ABSTRACT
Candida albicans, the most common human pathogenic fungus, can establish a persistent lethal infection in the intestine of the microscopic nematode Caenorhabditis elegans. The C. elegans-C. albicans infection model was previously adapted to screen for antifungal compounds. Modifications to this screen have been made to facilitate a high-throughput assay including co-inoculation of nematodes with C. albicans and instrumentation allowing precise dispensing of worms into assay wells, eliminating two labor-intensive steps. This high-throughput method was utilized to screen a library of 3,228 compounds represented by 1,948 bioactive compounds and 1,280 small molecules derived via diversity-oriented synthesis. Nineteen compounds were identified that conferred an increase in C. elegans survival, including most known antifungal compounds within the chemical library. In addition to seven clinically used antifungal compounds, twelve compounds were identified which are not primarily used as antifungal agents, including three immunosuppressive drugs. This assay also allowed the assessment of the relative minimal inhibitory concentration, the effective concentration in vivo, and the toxicity of the compound in a single assay.

Show MeSH
Related in: MedlinePlus