Synergistic effect of clomipramine and amphotericin on Leishmania amazonensis promastigotes

The search for new therapies involves the use of drug combinations (F) effective for conditions with similar pathogen-host biology and with molecular targets present in the parasite, but absent in the host, such as trypanothione reductase in parasites of the genus Leishmania. An F inhibitor...

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Detalles Bibliográficos
Autores principales: Barrionuevo, CN, Dimmer , JA, Tempesti , T, Rivarola, HW
Formato: Artículo revista
Lenguaje:Español
Publicado: Universidad Nacional Córdoba. Facultad de Ciencias Médicas. Secretaria de Ciencia y Tecnología 2023
Materias:
leishmania
Amphotericin
clomipramine
treatment
combination
anfotericina
clomipramina
tratamiento
combinación
Acceso en línea:https://revistas.unc.edu.ar/index.php/med/article/view/42666
Aporte de:
Revista de la Facultad de Ciencias Médicas de Córdoba de Universidad Nacional de Córdoba
id I10-R327-article-42666
record_format ojs
institution Universidad Nacional de Córdoba
institution_str I-10
repository_str R-327
container_title_str Revista de la Facultad de Ciencias Médicas de Córdoba
language Español
format Artículo revista
topic leishmania
Amphotericin
clomipramine
treatment
combination
leishmania
anfotericina
clomipramina
tratamiento
combinación
spellingShingle leishmania
Amphotericin
clomipramine
treatment
combination
leishmania
anfotericina
clomipramina
tratamiento
combinación
Barrionuevo, CN
Dimmer , JA
Tempesti , T
Rivarola, HW
Synergistic effect of clomipramine and amphotericin on Leishmania amazonensis promastigotes
topic_facet leishmania
Amphotericin
clomipramine
treatment
combination
leishmania
anfotericina
clomipramina
tratamiento
combinación
author Barrionuevo, CN
Dimmer , JA
Tempesti , T
Rivarola, HW
author_facet Barrionuevo, CN
Dimmer , JA
Tempesti , T
Rivarola, HW
author_sort Barrionuevo, CN
title Synergistic effect of clomipramine and amphotericin on Leishmania amazonensis promastigotes
title_short Synergistic effect of clomipramine and amphotericin on Leishmania amazonensis promastigotes
title_full Synergistic effect of clomipramine and amphotericin on Leishmania amazonensis promastigotes
title_fullStr Synergistic effect of clomipramine and amphotericin on Leishmania amazonensis promastigotes
title_full_unstemmed Synergistic effect of clomipramine and amphotericin on Leishmania amazonensis promastigotes
title_sort synergistic effect of clomipramine and amphotericin on leishmania amazonensis promastigotes
description The search for new therapies involves the use of drug combinations (F) effective for conditions with similar pathogen-host biology and with molecular targets present in the parasite, but absent in the host, such as trypanothione reductase in parasites of the genus Leishmania. An F inhibitor of this enzyme is clomipramine (Clo), used for its antidepressant action. The commercial formulations of the F are associated with excipients, which can be eliminated by simple methods and thus achieve their purification. The objective of this work was focused on analyzing the "in vitro" combined therapy of Amphotericin-B (Anf) and purified Clo on L. amazonensis promastigotes. Purification of Clo was carried out taking into account its high solubility in dichloromethane. 3x106 promastigotes/mL were incubated with concentrations of F alone (n=3) and combined (n=3) (Anf=0.02-0.62 μg/mL and Clo=0.48-15.6 μg/mL ). After 72 h, the parasite count was performed and the 50% inhibitory concentration (IC50) and the Combination Index (CI: antagonistic IC>1; IC=1 additive; IC<1 synergistic) were determined. Likewise, the isobologram corresponding to the IC50 values ​​of each of the F and their combinations was performed. The comparison of the treatments was made with an ANOVA and the Bonferroni post hoc test. The IC50 values ​​of each F were: Anf=0.28 μg/ml and Chlo=1.27 μg/ml, being significantly lower than the cytotoxic concentration in Vero Cells. Of the concentrations studied, the combination of Chlo with 0.02 (IC50=1.11 μg/ml); 0.04 (IC50=0.80 μg/ml) and 0.08 (IC50=0.78 μg/ml) of Anf were below the additivity line, evidencing a synergistic effect. These data coincided with the IC values ​​obtained for each concentration respectively: 0.95; 0.78 and 0.89. The use of purified F improved the antiparasitic effect allowing the reduction of Chlo concentration. The synergistic effect observed is consistent with the results found in previous studies on Trypanosoma cruzi and L. braziliensis.
publisher Universidad Nacional Córdoba. Facultad de Ciencias Médicas. Secretaria de Ciencia y Tecnología
publishDate 2023
url https://revistas.unc.edu.ar/index.php/med/article/view/42666
work_keys_str_mv AT barrionuevocn synergisticeffectofclomipramineandamphotericinonleishmaniaamazonensispromastigotes
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AT tempestit synergisticeffectofclomipramineandamphotericinonleishmaniaamazonensispromastigotes
AT rivarolahw synergisticeffectofclomipramineandamphotericinonleishmaniaamazonensispromastigotes
AT barrionuevocn efectosinergicodeclomipraminayanfotericinasobrepromastigotesdeleishmaniaamazonensis
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AT tempestit efectosinergicodeclomipraminayanfotericinasobrepromastigotesdeleishmaniaamazonensis
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first_indexed 2024-09-03T21:04:45Z
last_indexed 2024-09-03T21:04:45Z
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spelling I10-R327-article-426662023-10-19T21:20:08Z Synergistic effect of clomipramine and amphotericin on Leishmania amazonensis promastigotes Efecto sinérgico de clomipramina y anfotericina sobre promastigotes de Leishmania amazonensis Barrionuevo, CN Dimmer , JA Tempesti , T Rivarola, HW leishmania Amphotericin clomipramine treatment combination leishmania anfotericina clomipramina tratamiento combinación The search for new therapies involves the use of drug combinations (F) effective for conditions with similar pathogen-host biology and with molecular targets present in the parasite, but absent in the host, such as trypanothione reductase in parasites of the genus Leishmania. An F inhibitor of this enzyme is clomipramine (Clo), used for its antidepressant action. The commercial formulations of the F are associated with excipients, which can be eliminated by simple methods and thus achieve their purification. The objective of this work was focused on analyzing the "in vitro" combined therapy of Amphotericin-B (Anf) and purified Clo on L. amazonensis promastigotes. Purification of Clo was carried out taking into account its high solubility in dichloromethane. 3x106 promastigotes/mL were incubated with concentrations of F alone (n=3) and combined (n=3) (Anf=0.02-0.62 μg/mL and Clo=0.48-15.6 μg/mL ). After 72 h, the parasite count was performed and the 50% inhibitory concentration (IC50) and the Combination Index (CI: antagonistic IC>1; IC=1 additive; IC<1 synergistic) were determined. Likewise, the isobologram corresponding to the IC50 values ​​of each of the F and their combinations was performed. The comparison of the treatments was made with an ANOVA and the Bonferroni post hoc test. The IC50 values ​​of each F were: Anf=0.28 μg/ml and Chlo=1.27 μg/ml, being significantly lower than the cytotoxic concentration in Vero Cells. Of the concentrations studied, the combination of Chlo with 0.02 (IC50=1.11 μg/ml); 0.04 (IC50=0.80 μg/ml) and 0.08 (IC50=0.78 μg/ml) of Anf were below the additivity line, evidencing a synergistic effect. These data coincided with the IC values ​​obtained for each concentration respectively: 0.95; 0.78 and 0.89. The use of purified F improved the antiparasitic effect allowing the reduction of Chlo concentration. The synergistic effect observed is consistent with the results found in previous studies on Trypanosoma cruzi and L. braziliensis. La búsqueda de nuevas terapias involucra el uso de combinaciones de fármacos (F) efectivos para afecciones de similar biología patógeno-huésped y con blancos moleculares presentes en el parásito, pero ausentes en el huésped, como la tripanotiona reductasa en parásitos del género Leishmania. Un F inhibidor de esta enzima es la clomipramina (Clo), empleado por su acción antidepresiva. Las formulaciones comerciales de los F traen asociadas excipientes, que pueden ser eliminados mediante métodos sencillos y lograr así su purificación. El objetivo de este trabajo se centró en analizar la terapia combinada “in vitro” de Anfotericina-B (Anf) y Clo purificada sobre promastigotes de L. amazonensis. Se realizó la purificación de Clo teniendo en cuenta su alta solubilidad en diclorometano. Se incubaron 3x106 promastigotes/mL con concentraciones de los F solos (n=3) y combinados (n=3) (Anf=0,02-0,62 μg/mL y Clo=0,48-15,6 μg/mL). Luego de 72 h, se realizó el recuento de parásitos y se determinó la concentración inhibitoria del 50% (CI50) y el Índice de Combinación (IC: IC>1 antagónico; IC=1 aditivo; IC<1 sinérgico). Asimismo, se realizó el isobolograma correspondiente a los valores de CI50 de cada uno de los F y de sus combinaciones.  La comparación de los tratamientos se realizó con un ANOVA y el test post hoc de Bonferroni. Los valores de CI50 de cada F fueron: Anf=0,28 μg/ml y Clo=1,27 μg/ml, siendo significativamente menores que la concentración citotóxica en Células Vero. De las concentraciones estudiadas, la combinación de Clo con 0,02 (CI50=1,11 μg/ml); 0,04 (CI50=0,80 μg/ml) y 0,08 (CI50=0,78 μg/ml) de Anf se ubicaron por debajo de la línea de aditividad, evidenciando un efecto sinérgico. Estos datos fueron coincidentes con los valores de IC obtenidos para cada concentración respectivamente: 0,95; 0,78 y 0,89. La utilización del F purificado mejoró el efecto antiparasitario permitiendo reducir la concentración de Clo. El efecto sinérgico observado es coincidente con los resultados hallados en estudios previos en Trypanosoma cruzi y L. braziliensis. Universidad Nacional Córdoba. Facultad de Ciencias Médicas. Secretaria de Ciencia y Tecnología 2023-10-19 info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion application/pdf https://revistas.unc.edu.ar/index.php/med/article/view/42666 Revista de la Facultad de Ciencias Médicas de Córdoba.; Vol. 80 (2023): Suplemento JIC XXIV Revista de la Facultad de Ciencias Médicas de Córdoba; Vol. 80 (2023): Suplemento JIC XXIV Revista da Faculdade de Ciências Médicas de Córdoba; v. 80 (2023): Suplemento JIC XXIV 1853-0605 0014-6722 spa https://revistas.unc.edu.ar/index.php/med/article/view/42666/42862 Derechos de autor 2023 Universidad Nacional de Córdoba http://creativecommons.org/licenses/by-nc/4.0

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