Target Identification and Mechanism of Action of Picolinamide and Benzamide Chemotypes with Antifungal Properties

Cell Chem Biol. 2018 Mar 15;25(3):279-290.e7. doi: 10.1016/j.chembiol.2017.12.007. Epub 2018 Jan 4.

Abstract

Invasive fungal infections are accompanied by high mortality rates that range up to 90%. At present, only three different compound classes are available for use in the clinic, and these often suffer from low bioavailability, toxicity, and drug resistance. These issues emphasize an urgent need for novel antifungal agents. Herein, we report the identification of chemically versatile benzamide and picolinamide scaffolds with antifungal properties. Chemogenomic profiling and biochemical assays with purified protein identified Sec14p, the major phosphatidylinositol/phosphatidylcholine transfer protein in Saccharomyces cerevisiae, as the sole essential target for these compounds. A functional variomics screen identified resistance-conferring residues that localized to the lipid-binding pocket of Sec14p. Determination of the X-ray co-crystal structure of a Sec14p-compound complex confirmed binding in this cavity and rationalized both the resistance-conferring residues and the observed structure-activity relationships. Taken together, these findings open new avenues for rational compound optimization and development of novel antifungal agents.

Keywords: Sec14p; antifungal; benzamide; chemogenomics; co-crystal; functional variomics; lipid-transfer protein; picolinamide; target identification.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amides / chemistry
  • Amides / metabolism
  • Amides / pharmacology
  • Amino Acid Sequence
  • Antifungal Agents / chemistry
  • Antifungal Agents / metabolism*
  • Antifungal Agents / pharmacology
  • Aspergillus / drug effects
  • Benzamides / chemistry*
  • Benzamides / metabolism
  • Benzamides / pharmacology
  • Binding Sites
  • Candida albicans / drug effects
  • Crystallography, X-Ray
  • Drug Resistance, Fungal / drug effects
  • Microbial Sensitivity Tests
  • Molecular Dynamics Simulation
  • Mutagenesis, Site-Directed
  • Phospholipid Transfer Proteins / chemistry
  • Phospholipid Transfer Proteins / genetics
  • Phospholipid Transfer Proteins / metabolism
  • Picolinic Acids / chemistry*
  • Picolinic Acids / metabolism
  • Picolinic Acids / pharmacology
  • Protein Structure, Tertiary
  • Saccharomyces cerevisiae / drug effects
  • Saccharomyces cerevisiae / metabolism
  • Saccharomyces cerevisiae Proteins / chemistry
  • Saccharomyces cerevisiae Proteins / genetics
  • Saccharomyces cerevisiae Proteins / metabolism
  • Sequence Alignment
  • Structure-Activity Relationship

Substances

  • Amides
  • Antifungal Agents
  • Benzamides
  • Phospholipid Transfer Proteins
  • Picolinic Acids
  • Saccharomyces cerevisiae Proteins
  • picolinamide