Inhibitor Identification

Jadwiga Kaleta, Richele Severino

 

 Background and Goals

  • Cysteine proteases are implicated in    several diseases for which there is no effective available treatment.
  • Clinical trials for inhibitors of cathepsin K in the treatment of bone resorption  are currently ongoing.
  • The need to discover and develop novel small molecules of cysteine protease inhibitors is evident by significant interest of pharmaceutical industry.
  • The goal is to discover novel, potent cysteine protease inhibitors.
  • Soil bacteria and marine organisms are still unexploited as possible sources of novel molecules.

 

 

Methodology

  • Establish and optimize a 96-well plate high-throughput assay for cathepsin K inhibition as a first line of screening.
  • Use synthetic fluorogenic substrates (AMC flourophore, ex.380nm, em. 460nm) to detect  active site-directed inhibitors in a Spectra Max Gemini XS fluorimetric plate reader.
  • Optimize conditions to allow screening of crude and purified material in an efficient and reproducible way.
  • Optimize conditions to reduce non-specific hits by utilizing several blanks and cross-selectivity assays.