Ivermectin and curcumin cause plasma membrane rigidity in Leishmania amazonensis due to oxidative stress

Abstract

Spin label electron paramagnetic resonance (EPR) spectroscopy was used to study the mechanisms of action of ivermectin and curcumin against Leishmania (L.) amazonensis promastigotes. EPR spectra showed that treatment of the parasites with both compounds results in plasma membrane rigidity due to oxidative processes. With the IC₅₀ and EPR measurements for assays using different parasite concentrations, estimations could be made for the membrane-water partition coefficient (K_M/W), and the concentration of the compound in the membrane (c_m50) and in the aqueous phase (c_w50), which inhibits cell growth by 50%. The K_M/W values indicated that ivermectin has a greater affinity than curcumin for the parasite membrane. Therefore, the activity of ivermectin was higher for experiments with low cell concentrations, but for concentrations greater than 1.5 × 10⁸ parasites/mL the compounds did not show significantly different results. The c_m50 values indicated that the concentration of compound in the membrane leading to growth inhibition or membrane alteration is approximately 1 M for both ivermectin and curcumin. This high membrane concentration suggests that many ivermectin molecules per chlorine channel are needed to cause an increase in chlorine ion influx.

Keywords: Curcumin; Drug repositioning; Ivermectin; Leishmania; electron paramagnetic resonance.