Drug repurposing strategy against Trypanosoma cruzi infection: In vitro and in vivo assessment of the activity of metronidazole in mono- and combined therapy

Simões-Silva, Marianne Rocha; Araújo, Julianna Siciliano de; Oliveira, Gabriel Melo de; Demarque, Kelly Cristina; Peres, Raiza Brandão; D'Almeida-Melo, I.; Batista, D. G. J.; Silva, C.F. Da; Cardoso-Santos, Camila; Silva, P.B. Da; Batista, Marcos Meuser; Bahia, Maria T.; Soeiro, M. N. C.

doi:10.1016/j.bcp.2017.08.025

“…Other compounds have been identified by drug repositioning as possible trypanocidal drugs using different targets; such as the cruzipain (56–59), some T. cruzi kinases (60–62) and the trans-sialidase (63), among other examples. Other drug repurposing approaches have inferred the possible trypanocidal activity of already-approved drugs based on the reported effect or mechanism of action of some compounds in other organisms (64–69).…”

Section: Discussionmentioning

confidence: 99%

Crystal violet structural analogues identified by in silico drug repositioning present anti-Trypanosoma cruzi activity through inhibition of proline transporter TcAAAP069

Sayé

¹

,

Gauna

²

,

Valera-Vera

³

et al. 2019

Preprint

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BackgroundCrystal violet (CV) was used for several years in blood banks to eliminate the parasite Trypanosoma cruzi in endemic areas in order to prevent transfusion-transmitted Chagas disease. One mechanism of action described for CV involves inhibition of proline uptake. In T. cruzi, proline is essential for host cell infection and intracellular differentiation among other processes, and can be obtained through the proline permease TcAAAP069.Methodology/Principal FindingsCV inhibited proline transporter TcAAAP069 and parasites overexpressing this permease were 47-fold more sensitive to this compound than control parasites. Using CV as reference molecule, loratadine, cyproheptadine, olanzapine and clofazimine were identified as structurally related compounds to CV (structural analogues) by in silico drug repurposing through a similarity-based virtual screening protocol. All these already-approved drugs for clinical use inhibited TcAAAP069 activity with different efficacies and also presented trypanocidal action in epimastigotes, trypomastigotes and amastigotes of the Y strain. Additionally, loratadine, cyproheptadine and clofazimine showed trypanocidal effect on epimastigotes of the CL Brener and DM28c strains. Finally, a synergistic effect between benznidazole and the CV chemical analogues was evidenced by combination and dose-reduction indexes values in epimastigotes of the Y strain.Conclusions/SignificanceLoratadine, cyproheptadine and clofazimine inhibit TcAAAP069 proline transporter and also present trypanocidal effect against all T. cruzi life stages. These CV structural analogues could be a starting point to design therapeutic alternatives to treat Chagas disease by finding new indications for old drugs. This approach, called drug repurposing is a recommended strategy by the World Health Organization to treat neglected diseases, like Chagas disease, and combination therapy may improve the possibility of success of repositioned drugs.Author summaryChagas disease, caused by the parasite Trypanosoma cruzi, affects 7 million people worldwide. Despite there are two drugs available since 50 years ago, the therapy present severe side effects and is not effective in the chronic phase of the disease were most of the patients are diagnosed. Crystal violet (CV) was utilized as additive in blood banks to prevent transfusion-transmitted Chagas disease. Proline is involved in many pathways, like infection establishment and life cycle progression. In this work we first demonstrate that CV has the proline permease TcAAAP069 as one of its molecular targets. Then we search in a database of already-approved drugs for compounds that were structurally related to CV under the premise “similar structure, similar activity”. We identified three drugs that inhibit proline transport and present at least the same trypanocidal effect than benznidazole, the current treatment for Chagas disease. Finally we observed a synergistic effect with the multidrug combination therapy. Drug discovery is an expensive and time-consuming process and Chagas disease is associated with poverty. The discovery of new indications to old drugs, called drug repurposing, can facilitate a rapid and more profitable therapy application since preclinical trials and pharmacokinetic studies are already available.

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“…After 96 h of parasite infection, the supernatant, enriched with released trypomastigotes, was collected and the parasite purified following a previously established protocol (36). Bloodstream trypomastigotes (BT) of the Y strain were obtained from blood samples of infected albino Swiss mice at the peak of parasitemia, and the purified parasites were resuspended in RPMI medium as reported previously (38). Intracellular amastigotes of strains Y and Tulahuen were also evaluated.…”

Section: Methodsmentioning

confidence: 99%

“…Uninfected and T. cruzi-infected cultures subjected to only vehicle and BZ exposure were run in parallel. The results are expressed as percentages of parasite growth inhibition in compound-tested cells compared to that in infected cells and untreated cells (38). Triplicate samples were run in the same plate, and at least two assays were performed for each analysis.…”

Section: Methodsmentioning

confidence: 99%

“…Statistical analysis was performed individually for each assay by using an analysis of variance program, and the level of significance was set at a P value of Յ0.05. Interactions were categorized as described by Simões-Silva et al (38), with synergism classified by a mean ΑFICI of Յ0.5, antagonism classified by a mean ΑFICI of Ͼ4.0, and indifference (or an additive effect) classified by a mean ΑFICI of Ͼ0.5 and Յ4.0. All assays were conducted in triplicate, and the results are averages for at least three independent experiments (38).…”

Section: Methodsmentioning

confidence: 99%

See 1 more Smart Citation

Repurposing Strategy of Atorvastatin against Trypanosoma cruzi: In Vitro Monotherapy and Combined Therapy with Benznidazole Exhibit Synergistic Trypanocidal Activity

Araújo-Lima

¹

,

Peres

²

,

Silva

³

et al. 2018

Antimicrob Agents Chemother

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Statins are inhibitors of cholesterol synthesis, but other biological properties, such as antimicrobial effects, have also been assigned to them, leading to their designation as pleiotropic agents. Our goal was to investigate the activity and selectivity of atorvastatin (AVA) against by using models, aiming for more effective and safer therapeutic options through drug repurposing proposals for monotherapy and therapy in combination with benznidazole (BZ). Phenotypic screening was performed with different strains (Tulahuen [discrete typing unit {DTU} VI] and Y [DTU II]) and forms (intracellular forms, bloodstream trypomastigotes, and tissue-derived trypomastigotes) of the parasite. On assay of the Tulahuen strain, AVA was more active against intracellular amastigotes (selectivity index [SI] = 3). Also, against a parasite of another DTU (Y strain), this statin was more active (2.1-fold) and selective (2.4-fold) against bloodstream trypomastigotes (SI = 51) than against the intracellular forms (SI = 20). A cytomorphological approach using phalloidin-rhodamine permitted us to verify that AVA did not induced cell density reduction and that cardiac cells (CC) maintained their typical cytoarchitecture. Combinatory approaches using fixed-ratio methods showed that AVA and BZ gave synergistic interactions against both trypomastigotes and intracellular forms (mean sums of fractional inhibitory concentration indexes [∑FICIs] of 0.46 ± 0.12 and 0.48 ± 0.03, respectively). Thus, the repurposing strategy for AVA, especially in combination with BZ, which leads to a synergistic effect, is encouraging for future studies to identify novel therapeutic protocols for Chagas disease treatment.

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“…Other compounds have been identified by drug repositioning as possible trypanocidal drugs using different targets; such as the cruzipain [61][62][63][64], some T. cruzi kinases [65][66][67] and the trans-sialidase [68], among other examples. Other drug repurposing approaches have inferred the possible trypanocidal activity of already-approved drugs based on the reported effect or mechanism of action of some compounds in other organisms [69][70][71][72][73][74].…”

Section: Discussionmentioning

confidence: 99%

Crystal violet structural analogues identified by in silico drug repositioning present anti-Trypanosoma cruzi activity through inhibition of proline transporter TcAAAP069

Sayé

¹

,

Gauna

²

,

Valera-Vera

³

et al. 2020

View full text Add to dashboard Cite

Background Crystal violet (CV) was used for several years in blood banks to eliminate the parasite Trypanosoma cruzi in endemic areas in order to prevent transfusion-transmitted Chagas disease. One mechanism of action described for CV involves inhibition of proline uptake. In T. cruzi, proline is essential for host cell infection and intracellular differentiation among other processes, and can be obtained through the proline permease TcAAAP069. Methodology/Principal findings CV inhibited proline transporter TcAAAP069 and parasites overexpressing this permease were 47-fold more sensitive to this compound than control parasites. Using CV as reference molecule, loratadine, cyproheptadine, olanzapine and clofazimine were identified as structurally related compounds to CV (structural analogues) by in silico drug repurposing through a similarity-based virtual screening protocol. All these already-approved drugs for clinical use inhibited TcAAAP069 activity with different efficacies and also presented trypanocidal action in epimastigotes, trypomastigotes and amastigotes of the Y, CL Brener and Dm28c T. cruzi strains. Finally, a synergistic effect between benznidazole and the CV chemical analogues was evidenced by combination and dose-reduction indexes values in epimastigotes and trypomastigotes of the Y strain. Conclusions/Significance Loratadine, cyproheptadine and clofazimine inhibit TcAAAP069 proline transporter and also present trypanocidal effect against all T. cruzi life stages in strains from three different DTUs. These CV structural analogues could be a starting point to design therapeutic alternatives to treat Chagas disease by finding new indications for old drugs. This approach,

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Drug repurposing strategy against Trypanosoma cruzi infection: In vitro and in vivo assessment of the activity of metronidazole in mono- and combined therapy

Cited by 29 publications

References 39 publications

Crystal violet structural analogues identified by in silico drug repositioning present anti-Trypanosoma cruzi activity through inhibition of proline transporter TcAAAP069

Crystal violet structural analogues identified by in silico drug repositioning present anti-Trypanosoma cruzi activity through inhibition of proline transporter TcAAAP069

Repurposing Strategy of Atorvastatin against Trypanosoma cruzi: In Vitro Monotherapy and Combined Therapy with Benznidazole Exhibit Synergistic Trypanocidal Activity

Crystal violet structural analogues identified by in silico drug repositioning present anti-Trypanosoma cruzi activity through inhibition of proline transporter TcAAAP069

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