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In vitro anti-Leishmania activity of triclabendazole and its synergic effect with amphotericin B

INTRODUCTION: Leishmaniasis is a neglected tropical disease, with approximately 1 million new cases and 30,000 deaths reported every year worldwide. Given the lack of adequate medication for treating leishmaniasis, drug repositioning is essential to save time and money when searching for new therape...

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Autores principales: Borges, Beatriz Santana, Bueno, Gislayne de Paula, Tomiotto-Pellissier, Fernanda, Figueiredo, Fabiano Borges, Soares Medeiros, Lia Carolina
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9868945/
https://www.ncbi.nlm.nih.gov/pubmed/36699729
http://dx.doi.org/10.3389/fcimb.2022.1044665
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author Borges, Beatriz Santana
Bueno, Gislayne de Paula
Tomiotto-Pellissier, Fernanda
Figueiredo, Fabiano Borges
Soares Medeiros, Lia Carolina
author_facet Borges, Beatriz Santana
Bueno, Gislayne de Paula
Tomiotto-Pellissier, Fernanda
Figueiredo, Fabiano Borges
Soares Medeiros, Lia Carolina
author_sort Borges, Beatriz Santana
collection PubMed
description INTRODUCTION: Leishmaniasis is a neglected tropical disease, with approximately 1 million new cases and 30,000 deaths reported every year worldwide. Given the lack of adequate medication for treating leishmaniasis, drug repositioning is essential to save time and money when searching for new therapeutic approaches. This is particularly important given leishmaniasis’s status as a neglected disease. Available treatments are still far from being fully effective for treating the different clinical forms of the disease. They are also administered parenterally, making it challenging to ensure complete treatment, and they are extremely toxic, in some cases, causing death. Triclabendazole (TCBZ) is a benzimidazole used to treat fasciolosis in adults and children. It presents a lower toxicity profile than amphotericin B (AmpB) and is administered orally, making it an attractive candidate for treating other parasitoses. The mechanism of action for TCBZ is not yet well understood, although microtubules or polyamines could potentially act as a pharmacological target. TCBZ has already shown antiproliferative activity against T. cruzi, T. brucei, and L. infantum. However, further investigations are still necessary to elucidate the mechanisms of action of TCBZ. METHODS: Cytotoxicity assay was performed by MTT assay. Cell inhibition (CI) values were obtained according to the equation CI = (O.D treatment x 100/O.D. negative control). For Infection evaluation, fixated cells were stained with Hoechst and read at Operetta High Content Imaging System (Perkin Elmer). For growth curves, cell culture absorbance was measured daily at 600 nm. For the synergism effect, Fractional Inhibitory Concentrations (FICs) were calculated for the IC50 of the drugs alone or combined. Mitochondrial membrane potential (DYm), cell cycle, and cell death analysis were evaluated by flow cytometry. Reactive oxygen species (ROS) and lipid quantification were also determined by fluorimetry. Treated parasites morphology and ultrastructure were analyzed by electron microscopy. RESULTS: The selectivity index (SI = CC50/IC50) of TCBZ was comparable with AmpB in promastigotes and amastigotes of Leishmania amazonensis. Evaluation of the cell cycle showed an increase of up to 13% of cells concentrated in S and G2, and morphological analysis with scanning electron microscopy showed a high frequency of dividing cells. The ultrastructural analysis demonstrated large cytoplasmic lipid accumulation, which could suggest alterations in lipid metabolism. Combined administration of TCBZ and AmpB demonstrated a synergistic effect in vitro against intracellular amastigote forms with cSFICs of 0.25. CONCLUSIONS: Considering that TCBZ has the advantage of being inexpensive and administrated orally, our results suggest that TCBZ, combined with AmpB, is a promising candidate for treating leishmaniasis with reduced toxicity.
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spelling pubmed-98689452023-01-24 In vitro anti-Leishmania activity of triclabendazole and its synergic effect with amphotericin B Borges, Beatriz Santana Bueno, Gislayne de Paula Tomiotto-Pellissier, Fernanda Figueiredo, Fabiano Borges Soares Medeiros, Lia Carolina Front Cell Infect Microbiol Cellular and Infection Microbiology INTRODUCTION: Leishmaniasis is a neglected tropical disease, with approximately 1 million new cases and 30,000 deaths reported every year worldwide. Given the lack of adequate medication for treating leishmaniasis, drug repositioning is essential to save time and money when searching for new therapeutic approaches. This is particularly important given leishmaniasis’s status as a neglected disease. Available treatments are still far from being fully effective for treating the different clinical forms of the disease. They are also administered parenterally, making it challenging to ensure complete treatment, and they are extremely toxic, in some cases, causing death. Triclabendazole (TCBZ) is a benzimidazole used to treat fasciolosis in adults and children. It presents a lower toxicity profile than amphotericin B (AmpB) and is administered orally, making it an attractive candidate for treating other parasitoses. The mechanism of action for TCBZ is not yet well understood, although microtubules or polyamines could potentially act as a pharmacological target. TCBZ has already shown antiproliferative activity against T. cruzi, T. brucei, and L. infantum. However, further investigations are still necessary to elucidate the mechanisms of action of TCBZ. METHODS: Cytotoxicity assay was performed by MTT assay. Cell inhibition (CI) values were obtained according to the equation CI = (O.D treatment x 100/O.D. negative control). For Infection evaluation, fixated cells were stained with Hoechst and read at Operetta High Content Imaging System (Perkin Elmer). For growth curves, cell culture absorbance was measured daily at 600 nm. For the synergism effect, Fractional Inhibitory Concentrations (FICs) were calculated for the IC50 of the drugs alone or combined. Mitochondrial membrane potential (DYm), cell cycle, and cell death analysis were evaluated by flow cytometry. Reactive oxygen species (ROS) and lipid quantification were also determined by fluorimetry. Treated parasites morphology and ultrastructure were analyzed by electron microscopy. RESULTS: The selectivity index (SI = CC50/IC50) of TCBZ was comparable with AmpB in promastigotes and amastigotes of Leishmania amazonensis. Evaluation of the cell cycle showed an increase of up to 13% of cells concentrated in S and G2, and morphological analysis with scanning electron microscopy showed a high frequency of dividing cells. The ultrastructural analysis demonstrated large cytoplasmic lipid accumulation, which could suggest alterations in lipid metabolism. Combined administration of TCBZ and AmpB demonstrated a synergistic effect in vitro against intracellular amastigote forms with cSFICs of 0.25. CONCLUSIONS: Considering that TCBZ has the advantage of being inexpensive and administrated orally, our results suggest that TCBZ, combined with AmpB, is a promising candidate for treating leishmaniasis with reduced toxicity. Frontiers Media S.A. 2023-01-09 /pmc/articles/PMC9868945/ /pubmed/36699729 http://dx.doi.org/10.3389/fcimb.2022.1044665 Text en Copyright © 2023 Borges, Bueno, Tomiotto-Pellissier, Figueiredo and Soares Medeiros https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Cellular and Infection Microbiology
Borges, Beatriz Santana
Bueno, Gislayne de Paula
Tomiotto-Pellissier, Fernanda
Figueiredo, Fabiano Borges
Soares Medeiros, Lia Carolina
In vitro anti-Leishmania activity of triclabendazole and its synergic effect with amphotericin B
title In vitro anti-Leishmania activity of triclabendazole and its synergic effect with amphotericin B
title_full In vitro anti-Leishmania activity of triclabendazole and its synergic effect with amphotericin B
title_fullStr In vitro anti-Leishmania activity of triclabendazole and its synergic effect with amphotericin B
title_full_unstemmed In vitro anti-Leishmania activity of triclabendazole and its synergic effect with amphotericin B
title_short In vitro anti-Leishmania activity of triclabendazole and its synergic effect with amphotericin B
title_sort in vitro anti-leishmania activity of triclabendazole and its synergic effect with amphotericin b
topic Cellular and Infection Microbiology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9868945/
https://www.ncbi.nlm.nih.gov/pubmed/36699729
http://dx.doi.org/10.3389/fcimb.2022.1044665
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