Transport and local metabolism of budesonide and fluticasone propionate in a human bronchial epithelial cell line (Calu-3).
Calu-3 cells grown on microporous filters at an air interface for 16-18 days were incubated with the glucocorticosteroid (GCS) budesonide (BUD). Apparent permeability (P(app)) of BUD across Calu-3 cell monolayers was determined. Amount of BUD transported across Calu-3 cells was clearly concentration dependent, and no polarised transport was detected. When cells were loaded with BUD by incubation with drug solution (30 microM) for 2 h, 9.35 +/- 0.53% of the initial dose of BUD, corresponding to 5.5 microg BUD/mg cell protein was retained intracellularly, and released over a time period of 10 h. Apical release of BUD was significantly higher than release to the basolateral side of the monolayer. In comparison, when Calu-3 cells were loaded with fluticasone propionate (FP) solution (0.8 microM), about 20% of the initial dose of FP, corresponding to 0.3 microg FP/mg total cell protein content was detected intracellularly and released immediately (45 min). There was no difference in FP release between the apical and basolateral side of the cell layer. Mass spectrometry of cell extracts indicated the presence of fatty acid conjugates of BUD. We conclude that Calu-3 cells are able to store BUD by intracellular conjugation to fatty acids. We, therefore, suggest the use of the Calu-3 cell model as a tool for examination of local pharmacokinetics and metabolism of GCS at the bronchial epithelium.
Borchard
Gerrit
G
Leiden/Amsterdam Center for Drug Research, Division of Pharmaceutical Technology, P.O. Box 9502, The Netherlands. borchard@lacdr.leidenuniv.nl
Cassará
María Luz
ML
Roemelé
Peter E H
PE
Florea
Bogdan I
BI
Junginger
Hans E
HE
D000730:Androstadienes / Q000493:pharmacokinetics*; D001692:Biological Transport; D001980:Bronchi / Q000166:cytology / Q000378:metabolism*; D001993:Bronchodilator Agents / Q000493:pharmacokinetics*; D019819:Budesonide / Q000493:pharmacokinetics*; D002460:Cell Line; D004847:Epithelial Cells / Q000378:metabolism*; D000068298:Fluticasone; D006801:Humans
D000730:Androstadienes; D001993:Bronchodilator Agents; D019819:Budesonide; D000068298:Fluticasone