Highly selective inhibition of human CYP3Aa in vitro by azamulin and evidence that inhibition is irreversible.

Article Details

Citation

Stresser DM, Broudy MI, Ho T, Cargill CE, Blanchard AP, Sharma R, Dandeneau AA, Goodwin JJ, Turner SD, Erve JC, Patten CJ, Dehal SS, Crespi CL

Highly selective inhibition of human CYP3Aa in vitro by azamulin and evidence that inhibition is irreversible.

Drug Metab Dispos. 2004 Jan;32(1):105-12. doi: 10.1124/dmd.32.1.105.

PubMed ID
14709627 [ View in PubMed
]
Abstract

Azamulin [14-O-(5-(2-amino-1,3,4-triazolyl)thioacetyl)-dihydromutilin] is an azole derivative of the pleuromutilin class of antiinfectives. We tested the inhibition potency of azamulin toward 18 cytochromes P450 using human liver microsomes or microsomes from insect cells expressing single isoforms. In a competitive inhibition model, IC(50) values for CYP3A (0.03-0.24 microM) were at least 100-fold lower than all other non-CYP3A enzymes except CYP2J2 ( approximately 50-fold lower). The IC(50) value with heterologously expressed CYP3A4 was 15-fold and 13-fold less than those of CYP3A5 and CYP3A7, respectively. The reference inhibitor ketoconazole was less selective and exhibited potent inhibition (IC(50) values <10 microM) for CYP1A1, CYP1B1, CYP2B6, CYP2C8, CYP2C9, CYP2C19, CYP4F2, and CYP4F12. Inhibition of CYP3A by azamulin appeared sigmoidal and well behaved with the substrates 7-benzyloxy-4-trifluoromethylcoumarin, testosterone, and midazolam. Preincubation of 4.8 microM azamulin in the presence of NADPH for 10 min inhibited approximately 95% of testosterone 6beta-hydroxylase activity compared with preincubation in the absence of NADPH. Catalytic activities of CYP1A2, CYP2C8, CYP2C9, CYP2C19, CYP2D6, and CYP2E1 were unaffected by similar experiments. Incubation of azamulin with heterologously expressed CYP3A4 yielded a type I binding spectrum with a spectral dissociation constant of 3.5 microM, whereas no interaction was found with CYP2D6. Azamulin exhibited good chemical stability when stored in acetonitrile for up to 12 days. Aqueous solubility was found to be >300 microM. Azamulin represents an important new chemical tool for use in characterizing the contribution of CYP3A to the metabolism of xenobiotics.

DrugBank Data that Cites this Article

Drug Enzymes
DrugEnzymeKindOrganismPharmacological ActionActions
KetoconazoleCytochrome P450 1A1ProteinHumans
Unknown
Inhibitor
Inducer
Details
KetoconazoleCytochrome P450 1B1ProteinHumans
Unknown
Inhibitor
Details
KetoconazoleCytochrome P450 2B6ProteinHumans
Unknown
Inhibitor
Details
KetoconazoleCytochrome P450 2C8ProteinHumans
Unknown
Inhibitor
Details
KetoconazoleCytochrome P450 2C9ProteinHumans
Unknown
Inhibitor
Details
KetoconazoleCytochrome P450 4F12ProteinHumans
Unknown
Inhibitor
Details
KetoconazoleLeukotriene-B(4) omega-hydroxylase 1ProteinHumans
Unknown
Inhibitor
Details