An Integrated Approach for Identification and Target Validation of Antifungal Compounds Active against Erg11p

• DOI: 10.1128/aac.06332-11
• Publication Date: 2012-05-22T02:55:06Z
• Authors: Dominic Hoepfner; Shantanu Karkare; Stephen B. Helliwell; Martin Pfeifer; Markus Trunzer; Sophie De Bonnechose; Alfred Zimmerlin; Jianshi Tao; Daryl Richie; Andreas Hofmann; Stefan Reinker; Mathias Frederiksen; N. Rao Movva; Jeffrey A. Porter; Neil S. Ryder; Christian N. Parker
• Keywords: Azole; Docking (animal); Mechanism of Action; Antifungal drugs

Systemic life-threatening fungal infections represent a significant unmet medical need. Cell-based, phenotypic screening can be an effective means of discovering potential novel antifungal compounds, but it does not address target identification, normally required for compound optimization by medicinal chemistry. Here, we demonstrate combination screening, genetic, and biochemical approaches to identify characterize compounds. We isolated set non-azole compounds which no or mechanism action is known, using screen inhibition Saccharomyces cerevisiae proliferation. Haploinsufficiency profiling these in S. suggests that they Erg11p, cytochrome P450 family member, the azoles. Consistent with this, metabolic revealed buildup intermediates prior Erg11p activity, following treatment. Further, human also inhibited vitro assays modeled protein based on CYP51 crystal structure, silico docking interact heme center manner similar observations, Candida strains carrying azole-resistant alleles ERG11 are resistant this study. Thus, have identified inhibitors, systematic approach ligand characterization.