Prolonged anesthesia and decreased toxicity of enantiomeric-excess bupivacaine loaded in ionic gradient liposomes

Abstract

Bupivacaine is the most employed local anesthetic in surgical procedures, worldwide. Its systemic toxicity has directed the synthesis of the less toxic, S(-) enantiomer. This work describes a formulation of ionic gradient liposomes (IGL) containing _S75BVC, an enantiomeric excess mixture of 75% S(-) and 25% R(+) bupivacaine. IGL prepared with 250 mM (NH₄)₂SO₄ in the inner aqueous core of phosphatidylcholine and cholesterol (3:2 mol%) vesicles plus 0.5% _S75BVC showed average sizes of 312.5 ± 4.5 nm, low polydispersity (PDI < 0.18), negative zeta potentials (-14.2 ± 0.2 mV) and were stable for 360 days. The encapsulation efficiency achieved with IGL_S75BVC (%EE = 38.6%) was higher than with IGL prepared with racemic bupivacaine (IGL_RBVC, %EE = 28.3%). TEM images revealed spherical vesicles and µDSC analysis provided evidence on the interaction of the anesthetic with the lipid bilayer. Then, in vitro - release kinetics and cytotoxicity- and in vivo - toxic effects in Zebrafish and biochemical/histopathological analysis plus analgesia in Wistar rats - tests were performed. IGL_S75BVC exhibited negligible toxicity against Schwann cells and Zebrafish larvae, and it did not affect biochemical markers or the morphology of rat tissues (heart, brain, cerebellum, sciatic nerve). The in vitro release of _S75BVC from IGL was extended from 4 to 24 h, justifying the prolonged anesthetic effect measured in rats (~9 h). The advantages of IGL_S75BVC formulation over IGL_RBVC and plain bupivacaine formulations (prolonged anesthesia, preferential sensorial blockade, and no toxicity) confirm its potential for clinical use in surgical anesthesia.

Keywords: Bupivacaine; Drug delivery; Ionic gradient; Liposomes; Local anesthetics; Toxicology.