Peptidomimetic Vinyl Heterocyclic Inhibitors of Cruzain Effect Antitrypanosomal Activity

Abstract

Cruzain, an essential cysteine protease of the parasitic protozoan, <i>Trypanosoma cruzi</i>, is an important drug target for Chagas disease. We describe here a new series of reversible but time-dependent inhibitors of cruzain, composed of a dipeptide scaffold appended to vinyl heterocycles meant to provide replacements for the irreversible reactive "warheads" of vinyl sulfone inactivators of cruzain. Peptidomimetic vinyl heterocyclic inhibitors (PVHIs) containing Cbz-Phe-Phe/homoPhe scaffolds with vinyl-2-pyrimidine, vinyl-2-pyridine, and vinyl-2-(<i>N</i>-methyl)-pyridine groups conferred reversible, time-dependent inhibition of cruzain (<i>K</i>_i* = 0.1-0.4 μM). These cruzain inhibitors exhibited moderate to excellent selectivity versus human cathepsins B, L, and S and showed no apparent toxicity to human cells but were effective in cell cultures of <i>Trypanosoma brucei brucei</i> (EC₅₀ = 1-15 μM) and eliminated <i>T. cruzi</i> in infected murine cardiomyoblasts (EC₅₀ = 5-8 μM). PVHIs represent a new class of cruzain inhibitors that could progress to viable candidate compounds to treat Chagas disease and human sleeping sickness.