Evidence of significant contribution from CYP3A5 to hepatic drug metabolism.
CYP3A4 and CYP3A5 exhibit significant overlap in substrate specificity but can differ in product regioselectivity and formation activity. To further explore this issue, we compared the kinetics of product formation for eight different substrates, using heterologously expressed CYP3A4 and CYP3A5 and phenotyped human liver microsomes. Both enzymes displayed allosteric behavior toward six of the substrates. When it occurred, the "maximal" intrinsic clearance was used for quantitative comparisons. Based on this parameter, CYP3A5 was more active than CYP3A4 in catalyzing total midazolam hydroxylation (3-fold) and lidocaine demethylation (1.4-fold). CYP3A5 exhibited comparable metabolic activity as CYP3A4 (90-110%) toward dextromethorphan N-demethylation and carbamazepine epoxidation. CYP3A5-catalyzed erythromycin N-demethylation, total flunitrazepam hydroxylation, testosterone 6beta-hydroxylation, and terfenadine alcohol formation occurred with an intrinsic clearance that was less than 65% that of CYP3A4. Using two sets of human liver microsomes with equivalent CYP3A4-specific content but markedly different CYP3A5 content (group 1, predominantly CYP3A4; group 2, CYP3A4 + CYP3A5), we assessed the contribution of CYP3A5 to product formation rates determined at low substrate concentrations (< or = Km). Mean product formation rates for group 2 microsomes were 1.4- to 2.2-fold higher than those of group 1 (p < 0.05 for 5 of 8 substrates). After adjusting for CYP3A4 activity (itraconazole hydroxylation), mean product formation rates for group 2 microsomes were still significantly higher than those of group 1 (p < 0.05 for 3 substrates). We suggest that, under conditions when CYP3A5 content represents a significant fraction of the total hepatic CYP3A pool, the contribution of CYP3A5 to the clearance of some drugs may be an important source of interindividual variability.
Huang
Weili
W
Department of Pharmaceutics, Box 357610, University of Washington, Seattle, WA 98195-7610, USA.
Lin
Yvonne S
YS
McConn
Donavon J
DJ
Calamia
Justina C
JC
Totah
Rheem A
RA
Isoherranen
Nina
N
Glodowski
Mary
M
Thummel
Kenneth E
KE
D000818:Animals; D000935:Antifungal Agents / Q000378:metabolism; D016367:Baculoviridae; D001711:Biotransformation; D002460:Cell Line; D051544:Cytochrome P-450 CYP3A; D003577:Cytochrome P-450 Enzyme System / Q000096:biosynthesis / Q000378:metabolism*; D015786:Cytochromes b5 / Q000096:biosynthesis / Q000378:metabolism; D005445:Flunitrazepam / Q000378:metabolism; D006801:Humans; D066298:In Vitro Techniques; D007313:Insecta; D007527:Isoenzymes / Q000378:metabolism; D017964:Itraconazole / Q000378:metabolism; D007700:Kinetics; D008099:Liver / Q000201:enzymology / Q000378:metabolism*; D008862:Microsomes, Liver / Q000201:enzymology; D008874:Midazolam / Q000378:metabolism; D004364:Pharmaceutical Preparations / Q000378:metabolism*
D000935:Antifungal Agents; D007527:Isoenzymes; D004364:Pharmaceutical Preparations; D017964:Itraconazole; D005445:Flunitrazepam; D015786:Cytochromes b5; D003577:Cytochrome P-450 Enzyme System; C104464:CYP3A protein, human; C510164:CYP3A5 protein, human; D051544:Cytochrome P-450 CYP3A; C510163:CYP3A4 protein, human; D008874:Midazolam