Optimization of peptidyl allyl sulfones as clan CA cysteine protease inhibitors

Abstract

This research investigates the synthesis and inhibitory potency of a series of novel dipeptidyl allyl sulfones as clan CA cysteine protease inhibitors. The structure of the inhibitors consists of a R₁-Phe-R₂-AS-Ph scaffold (AS = allyl sulfone). R₁ was varied with benzyloxycarbonyl, morpholinocarbonyl, or N-methylpiperazinocarbonyl substituents. R₂ was varied with either Phe of Hfe residues. Synthesis involved preparation of vinyl sulfone analogues followed by isomerization to allyl sulfones using n-butyl lithium and t-butyl hydroperoxide. Sterics, temperature and base strength were all factors that affected the formation and stereochemistry of the allyl sulfone moiety. The inhibitors were assayed with three clan CA cysteine proteases (cruzain, cathepsin B and calpain I) as well as one serine protease (trypsin). The most potent inhibitor, (E)-Mu-Phe-Hfe-AS-Ph, displayed at least 10-fold selectivity for cruzain over clan CA cysteine proteases cathepsin B and calpain I with a k_obs/[I] of 6080 ± 1390 M⁻¹s⁻¹.

Keywords::

• Cruzain
• Chagas disease
• peptidyl vinyl sulfone

Acknowledgments

We would like to thank James McKerrow and his lab at the University of California − San Francisco for providing cruzain for enzyme inhibition analysis.

Declaration of interest

This material is based upon work supported by the National Science Foundation under Grant No. 0922775. The Howard Hughes Medical Institute provided additional research funding.