Identification and Characterization of Hundreds of Potent and Selective Inhibitors of Trypanosoma brucei Growth from a Kinase-Targeted Library Screening Campaign

In the interest of identification of new kinase-targeting chemotypes for target and pathway analysis and drug discovery in Trypanosomal brucei, a high-throughput screen of 42,444 focused inhibitors from the GlaxoSmithKline screening collection was performed against parasite cell cultures and counter-screened against human hepatocarcinoma (HepG2) cells. In this way, we have identified 797 sub-micromolar inhibitors of T. brucei growth that are at least 100-fold selective over HepG2 cells. Importantly, 242 of these hit compounds acted rapidly in inhibiting cellular growth, 137 showed rapid cidality. A variety of in silico and in vitro physicochemical and drug metabolism properties were assessed, and human kinase selectivity data were obtained, and, based on these data, we prioritized three compounds for pharmacokinetic assessment and demonstrated parasitological cure of a murine bloodstream infection of T. brucei rhodesiense with one of these compounds (NEU-1053). This work represents a successful implementation of a unique industrial-academic collaboration model aimed at identification of high quality inhibitors that will provide the parasitology community with chemical matter that can be utilized to develop kinase-targeting tool compounds. Furthermore these results are expected to provide rich starting points for discovery of kinase-targeting tool compounds for T. brucei, and new HAT therapeutics discovery programs.Author Summary: Human African trypanosomiasis, or sleeping sickness, affects 10,000 patients annually, yet current drugs for this disease are poor, with high toxicity and inconvenient dosing requirements. Trypanosoma brucei, the parasite that causes sleeping sickness, is sensitive to a class of compounds called kinase inhibitors, and our project was aimed at identifying kinase-targeting compounds that rapidly and irreversibly inhibit parasite growth. This was accomplished by high-throughput screening of over 42,000 compounds, which resulted in identification of 797 potent inhibitors of parasite growth that are non-toxic to human cells. These inhibitors were studied for the speed of their effects and reversibility of growth inhibition, and were grouped on the basis of chemical structure similarity. One compound was shown to cure mice from a bloodstream of infection of T. brucei. These compounds can now be utilized by the research community as starting points for new drug discovery, and also as tool compounds for understanding the function of kinases in T. brucei.