Ent-kaurane diterpenes isolated from n-hexane extract of Baccharis sphenophylla by bioactivity-guided fractionation target the acidocalcisomes in Trypanosoma cruzi

Costa-Silva, Thaís A.; Silva, Matheus L.; Antar, Guilherme Medeiros; Tempone, André G.; Lago, João Henrique G.

doi:10.1016/j.phymed.2021.153748

Cited by 5 publications

(13 citation statements)

References 27 publications

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“…Negative control consisted of untreated parasites and maximum intracellular calcium levels were obtained using 0.5% Triton X-100 (v/v). 36,37 Alterations in acidocalcisomes. Trypomastigotes (2 x 10 6 /well in 96 well plates) were washed with PBS and incubated with acridine orange dye (Thermo Fisher Scienti c) at 4 µM for 5 min in the dark.…”

Section: Mechanism Of Action Studiesmentioning

confidence: 99%

“…A luminometer (FilterMax F5 Multi-mode, Molecular Devices) was used for measuring luminescence intensity and ATP quantity calculated with an ATP standard curve. 37 Statistical analysis. Samples were tested in duplicates of three independent assays.…”

Section: Mechanism Of Action Studiesmentioning

confidence: 99%

“…Nigericin (4 µM) was used as positive control and non-treated parasites as negative control. 37 Mitochondrial membrane potential (Δψm). Trypomastigotes (2 x 10 6 /tube) were washed with HBSS supplemented with NaHCO3 and D-glucose and incubated with the mixture of compounds 1-4 (EC 50 value) at 37 o C and 5% CO 2 for 1 and 2 h. After washing, JC-1 dye (10 µM) was added and the parasites were incubated for 20 min at 37°C in the dark.…”

Section: Mechanism Of Action Studiesmentioning

confidence: 99%

“…Positive and negative control were CCCP (Sigma-Aldrich) at 100 µM and non-treated parasites, respectively. 37 Measurement of ROS levels. The mixture of compounds 1-4 was incubated with washed trypomastigotes (2 x 10 6 /well) in black 96 well plates in HBSS supplemented with NaHCO 3 (4.2 mM) and D-glucose (10 mM), at EC 50 value for 1 and 2 h (24 o C, 5% CO 2 ).…”

Section: Mechanism Of Action Studiesmentioning

confidence: 99%

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Enyne acetogenins from Porcelia macrocarpa fruit peels displayed anti-Trypanosoma cruzi activity in vitro and cause a reduction in the intracellular calcium level in the parasites

Thevenard

Brito

Costa-Silva

et al. 2023

Preprint

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Natural products are a promising source of new compounds with a wide spectrum of pharmacological properties, including antiprotozoal activities. Chagas disease, caused by the protozoan parasite Trypanosoma cruzi, is one of several neglected tropical diseases with reduced options for treatment, which presents limitations such as toxicity and ineffectiveness in the chronic stage of the disease. Aiming to investigate the Brazilian flora for the discovery of new anti-T. cruzi compounds, the MeOH extract from Porcelia macrocarpa R.E. Fries (Annonaceae) fruit peels displayed potent activity against trypomastigotes and intracellular amastigotes and was subjected to bioactivity-guided fractionation. Using different chromatographic steps, it was obtained a fraction composed of a mixture of four new chemically related acetogenins which were characterized as (2S*,3R*,4R*)-3-hydroxy-4-methyl-2-(n-octadeca-13’,17’-dien-11’-inil)butanolide (1), (2S*,3R*,4R*)-3-hydroxy-4-methyl-2-(n-eicosa-13’,19’-dien-11’-inil)butanolide (2), (2S*,3R*,4R*)-3-hydroxy-4-methyl-2-(n-octadec-13’-en-11’-inil)butanolide (3), and (2S*,3R*,4R*)-3-hydroxy-4-methyl-2-(n-eicosa-13’-en-11’-inil)butanolide (4) by analysis of NMR and UHPLC/ESI-HRMS data. The fraction composed of the mixture of compounds 1–4, displayed an EC50 of 4.9 and 2.5 mg/mL against trypomastigote and amastigote forms of T. cruzi, respectively, similar to standard drug benznidazole (EC50 of 4.8 and 1.4 mg/mL). Additionally, the fraction composed of 1–4 displayed no mammalian toxicity for murine fibroblasts (CC50 > 200 mg/mL), resulting in a SI > 40.8 and > 83.3 against trypomastigotes and amastigotes, respectively. Based on these results, the mechanism of action of this fraction was investigated. After a short-time incubation with the trypomastigotes, no alterations in the cell membrane permeability were observed. However, it was verified a decrease in the intracellular calcium of the parasites, without significant pH variations of the acidocalcisomes. The intracellular damages were followed by an upregulation of the reactive oxygen species (ROS) and ATP, but no depolarization effects were observed in the mitochondrial membrane potential. These data suggest that the fraction composed of 1–4 caused an irreversible oxidative stress in the parasites, leading to death. If adequately studied, these acetogenins can open new insights for discovery of new routes of death in T. cruzi.

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Section: Mechanism Of Action Studiesmentioning

confidence: 99%

Section: Mechanism Of Action Studiesmentioning

confidence: 99%

Section: Mechanism Of Action Studiesmentioning

confidence: 99%

Section: Mechanism Of Action Studiesmentioning

confidence: 99%

See 2 more Smart Citations

Enyne acetogenins from Porcelia macrocarpa fruit peels displayed anti-Trypanosoma cruzi activity in vitro and cause a reduction in the intracellular calcium level in the parasites

Thevenard

Brito

Costa-Silva

et al. 2023

Preprint

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show abstract

“…Parasites without drug treatments were included as a negative control. The data represent the mean of three independent assays, tested in duplicate [18].…”

Section: Evaluation Of Ca 2+ Levels In T Cruzi Trypomastigotesmentioning

confidence: 99%

New derivatives from dehydrodieugenol B and its methyl ether displayed high anti-Trypanosoma cruzi activity and cause depolarization of the plasma membrane and collapse the mitochondrial membrane potential

Galhardo

Ueno

Costa-Silva

et al. 2022

Chemico-Biological Interactions

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Enyne acetogenins from Porcelia macrocarpa displayed anti-Trypanosoma cruzi activity and cause a reduction in the intracellular calcium level

Thevenard

Brito

Costa-Silva

et al. 2023

Sci Rep

Self Cite

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Natural products are a promising source of new compounds with a wide spectrum of pharmacological properties, including antiprotozoal activities. Chagas disease, caused by the protozoan parasite Trypanosoma cruzi, is one of several neglected tropical diseases with reduced options for treatment, which presents limitations such as toxicity and ineffectiveness in the chronic stage of the disease. Aiming to investigate the Brazilian flora for the discovery of new anti-T. cruzi compounds, the MeOH extract from Porcelia macrocarpa R.E. Fries (Annonaceae) fruit peels displayed potent activity against trypomastigotes and intracellular amastigotes and was subjected to bioactivity-guided fractionation. Using different chromatographic steps, a fraction composed of a mixture of four new chemically related acetogenins was obtained. The compounds were characterized as (2S*,3R*,4R*)-3-hydroxy-4-methyl-2-(n-octadeca-13′,17′-dien-11′-inil)butanolide (1), (2S*,3R*,4R*)-3-hydroxy-4-methyl-2-(n-eicosa-13′,19′-dien-11′-inil)butanolide (2), (2S*,3R*,4R*)-3-hydroxy-4-methyl-2-(n-octadec-13′-en-11′-inil)butanolide (3), and (2S*,3R*,4R*)-3-hydroxy-4-methyl-2-(n-eicosa-13′-en-11′-inil)butanolide (4) by NMR analysis and UHPLC/ESI-HRMS data. The mixture of compounds 1–4, displayed an EC50 of 4.9 and 2.5 µg/mL against trypomastigote and amastigote forms of T. cruzi, respectively, similar to the standard drug benznidazole (EC50 of 4.8 and 1.4 µg/mL). Additionally, the mixture of compounds 1–4 displayed no mammalian toxicity for murine fibroblasts (CC50 > 200 µg/mL), resulting in a SI > 40.8 and > 83.3 against trypomastigotes and amastigotes, respectively. Based on these results, the mechanism of action of this bioactive fraction was investigated. After a short-time incubation with the trypomastigotes, no alterations in the cell membrane permeability were observed. However, it was verified a decrease in the intracellular calcium of the parasites, without significant pH variations of the acidocalcisomes. The intracellular damages were followed by an upregulation of the reactive oxygen species and ATP, but no depolarization effects were observed in the mitochondrial membrane potential. These data suggest that the mixture of compounds 1–4 caused an irreversible oxidative stress in the parasites, leading to death. If adequately studied, these acetogenins can open new insights for the discovery of new routes of death in T. cruzi.

show abstract

Ent-kaurane diterpenes isolated from n-hexane extract of Baccharis sphenophylla by bioactivity-guided fractionation target the acidocalcisomes in Trypanosoma cruzi

Cited by 5 publications

References 27 publications

Enyne acetogenins from Porcelia macrocarpa fruit peels displayed anti-Trypanosoma cruzi activity in vitro and cause a reduction in the intracellular calcium level in the parasites

Enyne acetogenins from Porcelia macrocarpa fruit peels displayed anti-Trypanosoma cruzi activity in vitro and cause a reduction in the intracellular calcium level in the parasites

New derivatives from dehydrodieugenol B and its methyl ether displayed high anti-Trypanosoma cruzi activity and cause depolarization of the plasma membrane and collapse the mitochondrial membrane potential

Enyne acetogenins from Porcelia macrocarpa displayed anti-Trypanosoma cruzi activity and cause a reduction in the intracellular calcium level

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