Calorimetric evidence of differentiated transport of limonin and nomilin through biomembranes

The effect exerted by two structurally similar limonoids possessing antifeedant and anticancer activity, limonin and nomilin, on the thermotropic behavior of model membranes constituted by dimyristoylphosphatidylcholine (DMPC) vesicles was studied by differential scanning calorimetry. Attention was directed to evaluate modifications in phytochemical-lipid interaction induced by compound structure and lipophilicity and to evidence their different membrane penetration. The two examined compounds, when dispersed in liposomes during their preparation, were found to exert a very different action on the LB-L, gel-to-liquid crystal phase transition of DMPC multilamellar vesicles. Nomilin caused a detectable effect on the transition temperature (T-m), shifting it toward lower values with a concomitant small decrease of the associated enthalpy (Delta H) changes, while limonin was not able to modify the lipid vesicles thermotropic behavior. Modifications induced by nomilin were a function of phytochemical concentration, while the different behavior of limonin can be due to the different polarity induced by the presence of the single A ring in nomilin that possesses an acetyl group versus the A,A' ring system of limonin. Solid limonoids and aqueous dispersions of multilamellar (MLVs) or unilamellar vesicles (LUVs) (limonoids molar fraction 0.045, 0.12, and 0.18) were left in touch for long incubation times at temperatures higher than T-m to detect their spontaneous transfer through the medium. By following this procedure, no interaction was detected for limonin with lipid vesicles. The rate of transfer and interaction of nomilin was a function of the kind of vesicle species (faster for LUV, slower for MLV). The interaction, monitored by compound transfer from the solid phytochemical to the lipidic species after several, periods of incubation, was on the same order as that detected by preparation carried out in organic solvent. The obtained results can be explained in terms of compound hydrophobicity, and a relation between compound structure and membrane interaction can be suggested. This allows the membrane interaction with nomilin, but the low water solubility of limonin hinders or totally blocks its transfer through the aqueous medium.