Direct interaction between substrates and endogenous steroids in the active site may change the activity of cytochrome P450 3A4.
CYP3A4 exhibits unusual kinetic characteristics that result from the metabolism of multiple substrate including endogenous steroids and some drugs that coexist at the active site. To clarify the mechanism of the effect of endogenous steroids on the drug metabolism, the interaction between substrates, nevirapine (NVP) and carbamazepine (CBZ), and endogenous steroids was investigated by theoretical calculations. When the activities of NVP 2-hydroxylation and CBZ 10,11-epoxidation by expressed CYP3A4 were measured in the presence of steroids, NVP 2-hydroxylation was found to be remarkably increased by aldosterone and inhibited by estradiol. CBZ 10,11-epoxidation was increased by androstenedione. Three-dimensional computer modeling has shown that the active site of CYP3A4 is especially large, permitting access of two substrate molecules. The interactions between NVP and aldosterone and between CBZ and androstenedione were estimated by theoretical calculations assuming the substrate and steroids to be present in the active site at the same time. It was shown that NVP or CBZ would be stably fixed close to the oxygen atom at the sixth ligand of heme by interaction with steroids, suggesting that NVP and CBZ may be hydroxylated more easily due to the interaction with steroids. Estradiol was also expected to interact with NVP via a pi/pi interaction between a benzene ring, in which the NVP hydroxylation site is located, and a benzene ring of estradiol, suggested to inhibit the reaction. From these results, interactions between the substrate and endogenous steroids in the active site may change the activity of CYP3A4.
Torimoto
Nao
N
Graduate School of Pharmaceutical Sciences, Chiba University, 1-33, Yayoi-cho, Inage-ku, Chiba 263-8522, Japan.
Ishii
Itsuko
I
Hata
Masayuki
M
Nakamura
Hiroyoshi
H
Imada
Hiroshi
H
Ariyoshi
Noritaka
N
Ohmori
Shigeru
S
Igarashi
Takashi
T
Kitada
Mitsukazu
M
D000328:Adult; D000450:Aldosterone / Q000737:chemistry* / Q000378:metabolism; D000735:Androstenedione / Q000737:chemistry* / Q000378:metabolism; D001665:Binding Sites / Q000235:genetics; D002220:Carbamazepine / Q000031:analogs & derivatives* / Q000737:chemistry; D051544:Cytochrome P-450 CYP3A; D003577:Cytochrome P-450 Enzyme System / Q000096:biosynthesis / Q000737:chemistry* / Q000235:genetics; D004795:Enzyme Stability / Q000235:genetics; D004958:Estradiol / Q000737:chemistry* / Q000378:metabolism; D006801:Humans; D006900:Hydroxylation; D007700:Kinetics; D009154:Mutation; D019829:Nevirapine / Q000737:chemistry; D012694:Serine / Q000235:genetics; D013379:Substrate Specificity / Q000235:genetics
D002220:Carbamazepine; D000735:Androstenedione; D012694:Serine; D000450:Aldosterone; D004958:Estradiol; D003577:Cytochrome P-450 Enzyme System; D019829:Nevirapine; C104464:CYP3A protein, human; D051544:Cytochrome P-450 CYP3A; C510163:CYP3A4 protein, human; C012227:carbamazepine epoxide