Investigation of dirithromycin and erythromycylamine uptake by human neutrophils in vitro.

Abstract

Dirithromycin, a new semisynthetic 14-membered-ring macrolide was avidly concentrated by human neutrophils in a time- but not concentration-dependent manner with mean cellular/extracellular, concentration ratios (C/E) of 9 within the first 5 min and up to 47 at 120 min. Erythromycylamine, the hydrolysis product of dirithromycin, was concentrated significantly less by neutrophils, reaching C/E values of 4 and 19 (at 5 and 120 min). A point of interest was the interindividual variability in the antibiotic uptake kinetics; in particular, 7 out of 47 neutrophil samples from different healthy volunteers displayed very slow uptake of both drugs (C/E values at 30 min: dirithromycin, 5.8; erythromycylamine, 4.6). The reason(s) for this is unknown. The uptake of both drugs was decreased at acidic pH and increased at basic pH. Chloroquine, an antimalarial drug which is concentrated in and alkalinizes azurophilic granules, reduced uptake by half. Metabolic inhibitors (2-4 dinitrophenol, sodium fluoride, potassium cyanide and sodium azide) did not impair the uptake of either drug but, interestingly, ouabain, an inhibitor of membrane Na+/K+ ATPase activity, impaired uptake by about 30%. Competitive inhibitors of some transport systems identified on neutrophil membrane (nucleosides, D-glucose and various aminoacids) did not alter the uptake of either drug. Dirithromycin and to a lesser extent, erythromycylamine, reached intracellular concentrations much higher than those required to inhibit the growth of sensitive microorganisms. Although the mechanism of uptake is not clear, one interesting hypothesis involves trapping by protonation into acidic compartments of neutrophils.