Insights of antiparasitic activity of sodium diethyldithiocarbamate against different strains of Trypanosoma cruzi

Abstract

Chagas disease is caused by Trypanosoma cruzi and affects thousands of people. Drugs currently used in therapy are toxic and have therapeutic limitations. In addition, the genetic diversity of T. cruzi represents an important variable and challenge in treatment. Sodium diethyldithiocarbamate (DETC) is a compound with pharmacological versatility acting as metal chelators and ROS generation. Thus, the objective was to characterize the antiparasitic action of DETC against different strains and forms of T. cruzi and their mechanism. The different strains of T. cruzi were grown in LIT medium. To evaluate the antiparasitic activity of DETC, epimastigote and trypomastigote forms of T. cruzi were used by resazurin reduction methods and by counting. Different response patterns were obtained between the strains and an IC50 of DETC ranging from 9.44 ± 3,181 to 60.49 ± 7.62 µM. Cell cytotoxicity against 3T3 and RAW cell lines and evaluated by MTT, demonstrated that DETC in high concentration (2222.00 µM) presents low toxicity. Yet, DETC causes mitochondrial damage in T. cruzi, as well as disruption in parasite membrane. DETC has antiparasitic activity against different genotypes and forms of T. cruzi, therefore, representing a promising molecule as a drug for the treatment of Chagas disease.

The repositioned drugs disulfiram/diethyldithiocarbamate combined to benznidazole: Searching for Chagas disease selective therapy, preventing toxicity and drug resistance

“…Both DETC and DSF express antiparasitic activities upon T. cruzi (Lane et al, 1996;de Freitas Oliveira et al, 2021;de Freitas Oliveira et al, 2022) and Leishmania (Khouri et al, 2010;Celes et al, 2016;Mazur et al, 2019). In macrophage cultures, DETC caused destruction of intracellular amastigotes with little effect on macrophage spreading pattern.…”

“…Interestingly, the T. brucei SOD gene sodb1 deletion increases sensitivity to NFX and BZ (Prathalingham et al, 2007) and SOD inhibition enhances NFX antitumoral activity (Koto et al, 2011). In this regard, the DSF and DETC strongly inhibit T. cruzi SOD (Giulivi et al, 1988), and is trypanocidal (de Freitas Oliveira et al, 2021), also exerting leishmanicidal effects (Khouri et al, 2010). Interestingly, the T. cruzi Fe-SOD is found within parasite mitochondria, preventing programmed cell death (Piacenza et al, 2007).…”

Design, synthesis, electrochemistry and anti-trypanosomatid hit/lead identification of nitrofuranylazines

“…For the single-point activity screening, the following equation was used to determine growth inhibition percentage: Growth inhibition% = 100 − [(ΔAbs sample − ΔAbs blank)/(ΔAbs neg control − ΔAbs blank) × 100] Compounds with growth inhibition of >70% qualified for further IC 50 evaluation. 69 To calculate the IC 50 , cell viability was determined using the following equation: Cell viability% = (ΔAbs sample − ΔAbs blank)/(ΔAbs neg control − ΔAbs blank) × 100 The IC 50 and Z -score were determined for each compound's three biological replicates using the cell viability% values and GraphPad Prism 5. The final IC 50 of each compound was determined by calculating the mean IC 50 , with standard deviation of the biological replicates.…”

“…Compounds with growth inhibition of >70% qualified for further IC 50 evaluation. 69 To calculate the IC 50 , cell viability was determined using the following equation:…”

“…cruzi antitrypomastigote assay. The activity of compounds against extracellular bloodstream forms (trypomastigotes) of T. cruzi were evaluated using an adapted method of de Freitas Oliveira et al 69 Cultured epimastigotes were centrifuged and resuspended in minimal essential medium (MEM) supplemented with 3% FBS for addition to a monolayer of cultured African green monkey kidney (Vero) cells (Cellonex, South Africa), which were maintained as indicated in the following cytotoxicity section. By changing the Vero culture media to MEM, the growth rate of the Vero cells is reduced, which in turn prevents reduced infection due to host cell proliferation.…”