Anti-Trypanosoma cruzi Activity of Metabolism Modifier Compounds

Abstract

Chagas disease is caused by the protozoan parasite Trypanosoma cruzi and affects over 6 million people worldwide. Development of new drugs to treat this disease remains a priority since those currently available have variable efficacy and frequent adverse effects, especially during the long regimens required for treating the chronic stage of the disease. T. cruzi modulates the host cell-metabolism to accommodate the cell cytosol into a favorable growth environment and acquire nutrients for its multiplication. In this study we evaluated the specific anti-T. cruzi activity of nine bio-energetic modulator compounds. Notably, we identified that 17-DMAG, which targets the ATP-binding site of heat shock protein 90 (Hsp90), has a very high (sub-micromolar range) selective inhibition of the parasite growth. This inhibitory effect was also highly potent (IC50 = 0.27 μmol L−1) against the amastigote intracellular replicative stage of the parasite. Moreover, molecular docking results suggest that 17-DMAG may bind T. cruzi Hsp90 homologue Hsp83 with good affinity. Evaluation in a mouse model of chronic T. cruzi infection did not show parasite growth inhibition, highlighting the difficulties encountered when going from in vitro assays onto preclinical drug developmental stages.

Novel Purine Chemotypes with Activity against Plasmodium falciparum and Trypanosoma cruzi

“…Vero cells were seeded in T-175 flasks (5 × 10 6 cells per flask) in DMEM supplemented with 1% penicillin–streptomycin and 10% FBS, and cultured for 24 h. Then, cells were washed once with PBS, and free-swimming trypomastigotes (1 × 10 7 trypomastigotes per flask; MOI = 1) in assay medium were added and allowed 18 h to infect [ 55 ]. After that, infected cell monolayers were washed with PBS and detached.…”

“…After that, infected cell monolayers were washed with PBS and detached. Cells were counted and diluted to a concentration of 5 × 10 5 cells per mL, before adding 100 µL per well to test plates already containing the drugs dispensed as described above [ 55 ]. In all cases, we included BNZ as control drug, and each plate contained its own negative (Vero cells and parasites) and positive (Vero cells) controls [ 55 ].…”

Amaryllidaceae plants: a potential natural resource for the treatment of Chagas disease

“…As previously described [ 29 ], Vero cells were infected in bulk. They were seeded in T-175 flasks (5 × 10 6 cells/flask) and cultured for 24 h. Then, cells were washed with PBS, and free-swimming trypomastigotes (1 × 10 7 trypomastigotes per flask; MOI = 1) were added for 18 h. Infected cell monolayers were washed in PBS and detached from the flask.…”

“…Cells were counted and diluted to a concentration of 5 × 10 5 cells/mL, before adding 100 µl per well to test plates already containing the extracts. In all cases we included BNZ as control drug, and each plate contained its own negative (Vero cells and parasites) and positive controls (Vero cells) [ 29 ].…”

Identification of compounds with activity against Trypanosoma cruzi within a collection of synthetic nucleoside analogs

“…After 18 h, infected Vero cells were PBS-washed and trypsin-detached, diluted at a concentration of 5×10 5 cells per ml in assay medium and 100 ml of that solution (50000 cells) added to 96-well test plates already containing the compounds. Plates were incubated at 37°C for another 72 h and read with CPRG, as described for the anti-T. cruzi assay (Martinez-Peinado et al, 2021a;Martinez-Peinado et al, 2021b;Martinez-Peinado et al, 2021c).…”