Evaluation of 4β-Hydroxycholesterol and 25-Hydroxycholesterol as Endogenous Biomarkers of CYP3A4: Study with CYP3A-Humanized Mice

被引：16

作者：

Nitta S.-I. ^{[1
,2
]}

Hashimoto M. ^{[1
]}

Kazuki Y. ^{[3
,4
]}

Takehara S. ^{[4
]}

Suzuki H. ^{[2
]}

Oshimura M. ^{[4
]}

Akita H. ^{[1
]}

Chiba K. ^{[1
]}

Kobayashi K. ^{[1
]}

机构：

[1] Laboratory of Pharmacology and Toxicology, Graduate School of Pharmaceutical Sciences, Chiba University, 1-8-1 Inohana, Chuo-ku, Chiba

[2] LSI Medience Corporation, 3-30-1 Shimura, Itabashi-ku, Tokyo

[3] Department of Biomedical Science, Institute of Regenerative Medicine and Biofunction, Graduate School of Medical Science, Tottori University, 86 Nishi-cho, Yonago, Tottori

[4] Chromosome Engineering Research Center (CERC), Tottori University, 86 Nishi-cho, Yonago, Tottori

来源：

The AAPS Journal | / 20卷 / 3期

基金：

日本学术振兴会;

关键词：

25-hydroxycholesterol; 4β-hydroxycholesterol; biomarker; CYP3A4; LC-MS/MS;

D O I：

10.1208/s12248-018-0186-9

中图分类号：

学科分类号：

摘要：

Cytochrome P450 3A (CYP3A) enzymes metabolize approximately half of all drugs on the market. Since the endogenous compounds 4β-hydroxycholesterol (4β-HC) and 25-hydroxycholesterol (25-HC) are generated from cholesterol via CYP3A enzymes, we examined whether the plasma levels of 4β-HC and 25-HC reflect hepatic CYP3A4 activity by using a CYP3A-humanized mouse model, in which the function of endogenous Cyp3a was genetically replaced by human CYP3A. CYP3A-humanized mice have great advantages for evaluation of the relationship between hepatic CYP3A protein levels and plasma and hepatic levels of 4β-HC and 25-HC. Levels of CYP3A4 protein in the liver microsomes of CYP3A-humanized mice were increased by treatment with pregnenolone-16α-carbonitrile, a CYP3A inducer. Hepatic and plasma levels of 4β-HC and 25-HC normalized by cholesterol were significantly correlated with hepatic CYP3A4 protein levels. In addition, in vitro studies using human liver microsomes showed that the formation of 4β-HC was strongly inhibited by a CYP3A inhibitor, while the inhibitory effect of the CYP3A inhibition on the formation of 25-HC was weak. These results suggested that CYP3A mainly contributed to the formation of 4β-HC in human liver microsomes, whereas other factors may be involved in the formation of 25-HC. In conclusion, the in vivo studies using CYP3A-humanized mice suggest that plasma 4β-HC and 25-HC levels reflect hepatic CYP3A4 activity. Furthermore, taking the results of in vitro studies using human liver microsomes into consideration, 4β-HC is a more reliable biomarker of hepatic CYP3A activity. © 2018, American Association of Pharmaceutical Scientists.

引用

共 48 条

[1]

Shimada T., Yamazaki H., Mimura M., Inui Y., Guengerich F.P., Interindividual variations in human liver cytochrome P-450 enzymes involved in the oxidation of drugs, carcinogens and toxic chemicals: studies with liver microsomes of 30 Japanese and 30 Caucasians, J Pharmacol Exp Ther, 270, 1, pp. 414-423, (1994)

[2]

Thummel K.E., Wilkinson G.R., In vitro and in vivo drug interactions involving human CYP3A, Annu Rev Pharmacol Toxicol, 38, 1, pp. 389-430, (1998)

[3]

Baker A.L., Kotake A.N., Schoeller D.A., Clinical utility of breath tests for the assessment of hepatic function, Semin Liver Dis, 3, 4, pp. 318-329, (1983)

[4]

Watkins P.B., Murray S.A., Winkelman L.G., Heuman D.M., Wrighton S.A., Guzelian P.S., Erythromycin breath test as an assay of glucocorticoid-inducible liver cytochromes P-450. Studies in rats and patients, J Clin Invest, 83, 2, pp. 688-697, (1989)

[5]

Watkins P.B., Hamilton T.A., Annesley T.M., Ellis C.N., Kolars J.C., Voorhees J.J., The erythromycin breath test as a predictor of cyclosporine blood levels, Clin Pharmacol Ther, 48, 2, pp. 120-129, (1990)

[6]

Thummel K.E., Shen D.D., Podoll T.D., Kunze K.L., Trager W.F., Hartwell P.S., Et al., Use of midazolam as a human cytochrome P450 3A probe: I. In vitro-in vivo correlations in liver transplant patients, J Pharmacol Exp Ther, 271, 1, pp. 549-556, (1994)

[7]

Thummel K.E., Shen D.D., Podoll T.D., Kunze K.L., Trager W.F., Bacchi C.E., Et al., Use of midazolam as a human cytochrome P450 3A probe: II. Characterization of inter- and intraindividual hepatic CYP3A variability after liver transplantation, J Pharmacol Exp Ther, 271, 1, pp. 557-566, (1994)

[8]

Thummel K.E., O'Shea D., Paine M.F., Shen D.D., Kunze K.L., Perkins J.D., Et al., Oral first-pass elimination of midazolam involves both gastrointestinal and hepatic CYP3A-mediated metabolism, Clin Pharmacol Ther, 59, 5, pp. 491-502, (1996)

[9]

Kinirons M.T., O'Shea D., Downing T.E., Fitzwilliam A.T., Joellenbeck L., Groopman J.D., Et al., Absence of correlations among three putative in vivo probes of human cytochrome P4503A activity in young healthy men, Clin Pharmacol Ther, 54, 6, pp. 621-629, (1993)

[10]

Bienvenu T., Rey E., Pons G., D'athis P., Olive G., A simple non-invasive procedure for the investigation of cytochrome P-450 IIIA dependent enzymes in humans, Int J Clin Pharmacol Ther Toxicol, 29, 11, pp. 441-445, (1991)

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