Local anesthetics destabilize lipid membranes by breaking hydration shell: infrared and calorimetry studies

Differential scanning calorimetry (DSC) showed that local anesthetics decreased the pretransition (L beta'-->P beta') temperature of dipalmitoylphosphatidylcholine (DPPC) vesicle membranes four- to five-fold more than the main transition (P beta'-->L alpha) temperature. Because pretransition is mainly a change in the hydrophilic head property (tilted-rippled), the stronger effect on the pretransition suggests that the primary action site of local anesthetics is the lipid-water interface. The interfacial effect was analyzed by Fourier-transform infrared spectroscopy (FTIR) in water-in-oil (CCl4) reversed micelles. FTIR showed that the local anesthetics released hydrogen-bonded water molecules from the phosphate (P = O bands) and glycerol (sn-2 C = O) moieties. The N-H stretching band of the local anesthetics was deconvoluted into two bands: hydrogen bonded to the phosphate moiety of the lipid and free (unbound to lipid). The formation constants between lipid P = O and anesthetic N-H were estimated in CCl4 from the spectral changes: 110 M-1 for lidocaine and 250 M-1 for dibucaine. This small difference in the formation constants cannot explain the ten-fold stronger effect on the phase-transition temperature of dibucaine over lidocaine. By comparing the local anesthetic adsorption to the air/water interface in the presence and absence of lipid monolayers, we have previously shown (Lin et al. (1980) Biochim. Biophys. Acta 598, 51-65) that lipid-anesthetics interaction involves three forces: lipophilic effect, hydrophobic effect, and anesthetic-anesthetic interaction. The anesthetic potency depends mainly on the hydrophobic effect (the difference in the standard molar free energies of local anesthetics in water and at the interface) and anesthetic-anesthetic interaction energy. The anesthetic-anesthetic interaction means cooperativity of local anesthetics for the interfacial density: local anesthetics condense at the membrane surface when there are enough anesthetic molecules present at the interface to attract more anesthetics. The present data suggest that anesthetic action is directed to the interface between water and macromolecule, whether it is lipid membranes or proteins.