In Vitro Assessment of Uptake and Lysosomal Sequestration of Respiratory Drugs in Alveolar Macrophage Cell Line NR8383

PURPOSE: To assess accumulation and lysosomal sequestration of 9 drugs used in respiratory indications (plus imipramine as positive control) in the alveolar macrophage (AM) cell line NR8383. METHODS: For all drugs, uptake at 5 μM was investigated at 37 and 4°C to delineate active uptake and passive diffusion processes. Accumulation of basic clarithromycin, formoterol and imipramine was also assessed over 0.1–100 μM concentration range. Lysosomal sequestration was investigated using ammonium chloride (NH(4)Cl), monensin and nigericin. Impact of lysosomal sequestration on clarithromycin accumulation kinetics was investigated. RESULTS: Both cell-to-medium concentration ratio (K(p)) and uptake clearance (CL(uptake)) ranged > 400-fold for the drugs investigated. The greatest K(p) was observed for imipramine (391) and clarithromycin (82), in contrast to no accumulation seen for terbutaline. A concentration-dependent accumulation was evident for the basic drugs investigated. Imipramine and clarithromycin K(p) and CL(uptake) were reduced by 59–85% in the presence of NH(4)Cl and monensin/nigericin, indicating lysosomal accumulation, whereas lysosomal sequestration was not pronounced for the other 8 respiratory drugs. Clarithromycin uptake rate was altered by NH(4)Cl, highlighting the impact of subcellular distribution on accumulation kinetics. CONCLUSIONS: This study provides novel evidence of the utility of NR8383 for investigating accumulation and lysosomal sequestration of respiratory drugs in AMs. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1007/s11095-015-1753-8) contains supplementary material, which is available to authorized users.