Identification of a new class of nonpeptidic inhibitors of cruzain

J Am Chem Soc. 2008 May 21;130(20):6404-10. doi: 10.1021/ja710254m. Epub 2008 Apr 25.


Cruzain is the major cysteine protease of Trypanosoma cruzi, which is the causative agent of Chagas disease and is a promising target for the development of new chemotherapy. With the goal of developing potent nonpeptidic inhibitors of cruzain, the substrate activity screening (SAS) method was used to screen a library of protease substrates initially designed to target the homologous human protease cathepsin S. Structure-based design was next used to further improve substrate cleavage efficiency by introducing additional binding interactions in the S3 pocket of cruzain. The optimized substrates were then converted to inhibitors by the introduction of cysteine protease mechanism-based pharmacophores. Inhibitor 38 was determined to be reversible even though it incorporated the vinyl sulfone pharmacophore that is well documented to give irreversible cruzain inhibition for peptidic inhibitors. The previously unexplored beta-chloro vinyl sulfone pharmacophore provided mechanistic insight that led to the development of potent irreversible acyl- and aryl-oxymethyl ketone cruzain inhibitors. For these inhibitors, potency did not solely depend on leaving group p K a, with 2,3,5,6-tetrafluorophenoxymethyl ketone 54 identified as one of the most potent inhibitors with a second-order inactivation constant of 147,000 s (-1) M (-1). This inhibitor completely eradicated the T. cruzi parasite from mammalian cell cultures and consequently has the potential to lead to new chemotherapeutics for Chagas disease.

Publication types

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

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Cathepsins / chemistry
  • Cathepsins / metabolism
  • Cathepsins / pharmacology
  • Coumarins / chemistry
  • Coumarins / metabolism
  • Cysteine Endopeptidases / chemistry
  • Cysteine Endopeptidases / metabolism
  • Cysteine Proteinase Inhibitors / chemistry*
  • Cysteine Proteinase Inhibitors / metabolism
  • Cysteine Proteinase Inhibitors / pharmacology*
  • Drug Design
  • Ketones / chemistry*
  • Ketones / metabolism
  • Ketones / pharmacology*
  • Macrophages / parasitology
  • Mice
  • Molecular Sequence Data
  • Protozoan Proteins / antagonists & inhibitors*
  • Protozoan Proteins / chemistry
  • Protozoan Proteins / metabolism
  • Sequence Alignment
  • Structure-Activity Relationship
  • Sulfones / chemistry
  • Sulfones / metabolism
  • Sulfones / pharmacology
  • Triazoles
  • Trypanosoma cruzi / drug effects
  • Trypanosoma cruzi / enzymology


  • Coumarins
  • Cysteine Proteinase Inhibitors
  • Ketones
  • Protozoan Proteins
  • Sulfones
  • Triazoles
  • divinyl sulfone
  • Cathepsins
  • Cysteine Endopeptidases
  • cruzain, Trypanosoma cruzi
  • cathepsin S
  • 7-amino-4-methylcoumarin