Comparison of CYP3A activities in a subclone of Caco-2 cells (TC7) and human intestine.
OBJECTIVE
To compare the activity of the CYP3A enzyme expressed by TC7, a cell culture model of the intestinal epithelial cell, to the activity of human intestinal CYP3A4, using terfenadine as a substrate.


METHODS
The metabolism of terfenadine was investigated in intact cells and microsomal preparations from TC7, human intestine, and liver. The effect of two CYP3A inhibitors, ketoconazole and troleandomycin (TAO), on the metabolism of terfenadine was also examined.


RESULTS
Only hydroxy-terfenadine was detected in TC7 microsomal incubations. In contrast, azacyclonol and hydroxy-terfenadine were detected in human intestinal and hepatic microsomal incubations. The Km values for hydroxy-terfenadine formation in TC7 cells, intestine and liver microsomes were 1.91, 2.5, and 1.8, microM respectively. The corresponding Vmax values were 2.11, 61.0, and 370 pmol/min/mg protein. Km values for azacyclonol in intestinal and hepatic samples were 1.44 and 0.82 microM and the corresponding Vmax values were 14 and 60 pmol/min/mg protein. The formation of hydroxy-terfenadine was inhibited by ketoconazole and TAO in human intestine and TC7 cell microsomes. The Km and Vmax values for terfenadine metabolism in intact TC7 cells were similar to those from TC7 cell microsomes.


CONCLUSIONS
Our results indicate that TC7 cells are a potentially useful alternative model for studies of CYP3A mediated drug metabolism. The CYP3A expressed by TC7 cells is not CYP3A4, but probably CYP3A5, making this cell line suitable for studies of colonic drug transport and metabolism.
Raeissi
S D
SD
Drug Metabolism and Pharmacokinetics, Rhone-Poulenc Rorer Central Research, Collegeville, Pennsylvania 19426-0107, USA.
Guo
Z
Z
Dobson
G L
GL
Artursson
P
P
Hidalgo
I J
IJ
D000906:Antibodies / Q000276:immunology; D001189:Aryl Hydrocarbon Hydroxylases; D018938:Caco-2 Cells; D051544:Cytochrome P-450 CYP3A; D065607:Cytochrome P-450 Enzyme Inhibitors; D003577:Cytochrome P-450 Enzyme System / Q000276:immunology / Q000378:metabolism*; D004791:Enzyme Inhibitors / Q000494:pharmacology; D006801:Humans; D006900:Hydroxylation; D007413:Intestinal Mucosa / Q000201:enzymology* / Q000648:ultrastructure; D007583:Jejunum / Q000201:enzymology / Q000648:ultrastructure; D007654:Ketoconazole / Q000494:pharmacology; D008861:Microsomes / Q000201:enzymology; D008862:Microsomes, Liver / Q000201:enzymology; D010084:Oxidation-Reduction; D010089:Oxidoreductases, N-Demethylating / Q000037:antagonists & inhibitors / Q000378:metabolism*; D013250:Steroid Hydroxylases / Q000276:immunology / Q000378:metabolism; D016593:Terfenadine / Q000378:metabolism / Q000493:pharmacokinetics; D014217:Troleandomycin / Q000494:pharmacology
D000906:Antibodies; D065607:Cytochrome P-450 Enzyme Inhibitors; D004791:Enzyme Inhibitors; D016593:Terfenadine; D003577:Cytochrome P-450 Enzyme System; D014217:Troleandomycin; D013250:Steroid Hydroxylases; D001189:Aryl Hydrocarbon Hydroxylases; D051544:Cytochrome P-450 CYP3A; C022657:steroid hormone 6-beta-hydroxylase; D010089:Oxidoreductases, N-Demethylating; D007654:Ketoconazole