In vitro biological assays. Leishmania infantum logarithmic phase promastigotes were incubated with b-AD (0–61.96 μM) or AmpB (0–1.08 μM) for 48 h at 24 °C. Cell viability was analyzed by the MTT method and the 50% Leishmania inhibitory concentration (IC50) was calculated by applying sigmoidal regression of dose-response curve. Murine macrophages were also incubated with b-AD (0–123.92 μM) or AmpB (0–10.82 μM) for 48 h at 37 °C in 5% CO2, and the 50% cell inhibitory concentration (CC50) was also evaluated by a dose-response curve. The selectivity index (SI) was calculated by the ratio between the CC50 and IC50 values. The 50% human red blood cells inhibitory concentration (RBC50) was determined by incubating the 5% red cell suspension with b-AD (0–123.92 μM) or AmpB (0–10.82 μM) for 1 h at 37 °C in 5% CO2. The lysis percentage was evaluated spectrophotometrically, and the absence or presence of hemolysis were determined by replacing b-AD for PBS or distilled water, respectively. Results are shown as mean ± standard deviation of the groups.
|Compound||IC50 (μM)||CC50 (μM)||SI||RBC50 (μM)|
|β-acetyl-digitoxin||20.94 ± 2.60||453.04 ± 23.29||21.64||334.95 ± 24.90|
|Amphotericin B||0.11 ± 0.03||0.86 ± 0.11||7.82||12.66 ± 1.52|
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