TITLE:
Kinetics of Local Anesthetic Interactions with Model Membranes: Adsorption and Permeation
AUTHORS:
Gary Strichartz
KEYWORDS:
Local Anesthetic, Fluorescence, Phospholipids, Kinetics, Stop-Flow Technique, Adsorption, Permeation
JOURNAL NAME:
Open Journal of Anesthesiology,
Vol.12 No.1,
January
30,
2022
ABSTRACT: Introduction: Local anesthetics (LAs) must be adsorbed to and permeate through membranes to reach their sites of action. The rates of these processes critically affect drug actions at the cellular level and also the in vivo pharmacokinetics of perineurally injected drugs. Therefore, we measured the adsorption equilibria and permeation kinetics of two LAs local anesthetics with markedly different hydrophobicities: tetracaine (TTC, Poctanol:buffer = 3.6 × 103) and procaine (PRO, Poctanol:buffer = 8.1 × 101), in unilamellar membranes of different composition and fluidity. Methods: Interactions with unilamellar vesicles made of dipalmitoylphosphatidylcholine (DPPC), at 23°C (solid gel state) or 45°C (liquid crystal state), without or with cholesterol (Chol), were investigated by a combination of fluorescence and stopped-flow techniques. Membrane adsorption was monitored by the LA’s fluorescence. Membrane permeation was monitored by the decrease of fluorescence intensity of intra-vesicular pyranine, when quenched by neighboring TTC or PRO. Results: Ionized TTC (TTCH+) and procaine (PROH+) adsorb to membranes more slowly and weakly than their neutral counterparts (time constant τ ~ 0.1 sec). Fluidizing of membranes, at the higher temperature (45°C) or by Chol inclusion, accelerated LA adsorption (faster than the mixing time of the stopped-flow device, 0.008 s). Permeation of protonated TTCH+ through the solid gel DPPC membranes (23°C) occurred far more slowly than adsorption, (τ = 36.7 ± 0.7 s, n = 9), and 3 times slower than neutral TTC (τ = 10.9 ± 0.7 s, n = 9); neutral PRO permeated these membranes at the same rate as TTC. Inclusion of Chol with DPPC, disordering the fatty acyl tails of membrane phospholipids while crowding their polar headgroups, slowed permeation of TTC and of PRO to an even greater degree. Conclusions: Local anesthetic permeation through membranes is limited by the transport across the membrane core and not by the initial binding. Drug ionization greatly slows permeation, but greater hydrophobicity does not facilitate it. Lipid crowding caused by Chol, a normal membrane component, slows permeation by disorienting the LA bound at the surface.