Antileishmanial activities of three chalcone derivatives and their association with plasma membrane rigidity as assessed by EPR spectroscopy

Novel chalcone-like compounds, LQFM332 (4), LQFM333 (5), and LQFM341 (6), were synthesized and their activity against Leishmania (L.) amazonensis was evaluated and compared. Compound 4 was synthesized from the molecular hybridization of vanillin (9) and butylated hydroxytoluene (BHT), forming a chalcone scaffold. BHT was replaced by another apocynin to give compound 5, while for compound 6 the Michael acceptor double bond from compound 4 was reduced. The IC50 values for compounds 4, 5, and 6 against the Leishmania parasite were 7, 20, and 16 & mu;M for the promastigote form, and 19, 11, and 1 & mu;M for the amastigote form inside infected macrophages, respectively. Electron paramagnetic resonance (EPR) spectroscopy of a lipid spin label incorporated into the parasite plasma membrane demonstrated that treatment with these compounds in culture medium causes membrane rigidity in a concentration-dependent manner, owing to oxidative processes. From the IC50 values with different cell concentrations, the membrane-water partition coefficient (KM/W) of the compounds could be estimated, as well as their concentrations in the membrane (cm50) and in the aqueous culture medium (cw50) that reduce antiproliferative activity by 50%. Values for these biophysical parameters were similar to those reported for the antileishmanial drug, miltefosine. KM/W values indicated that the compounds have high affinity for the parasite membrane, with the compound containing the BHT group having the highest affinity. Compound 5, which contained a two 4-OH-3-methoxybenzene scaffold, showed the lowest values of cw50 and cm50. Compound 6 showed that the reduction of the double bond decreased the membrane affinity and the cw50 value, but maintained the cm50, demonstrating the same activity as compound 4 for assays with higher concentration of cells. As cytotoxic activities were modulated by effects on the cell membrane, our results suggest that the primary action of the compounds is on the parasite membrane.

Automated summary

In this study, novel chalcone-like compounds were synthesized and evaluated for their activity against Leishmania (L.) amazonensis. The compounds showed varying potency against the parasite, with compound 4 being the most effective. Electron paramagnetic resonance spectroscopy revealed that the compounds induced membrane rigidity in a concentration-dependent manner. The compounds also exhibited high affinity for the parasite membrane, similar to the antileishmanial drug miltefosine.