Evaluation of microsomal incubation conditions on CYP3A4-mediated metabolism of cyclosporine a by a statistical experimental design

被引：10

作者：

Hermann, M ^{[1
]}

Kase, ET ^{[1
]}

Molden, E ^{[1
]}

Christensen, H ^{[1
]}

机构：

[1] Univ Oslo, Sch Pharm, Dept Pharmaceut Biosci, N-0316 Oslo, Norway

来源：

CURRENT DRUG METABOLISM | 2006年 / 7卷 / 03期

关键词：

microsomes; incubation conditions; experimental design; PLS; cyclosporine;

D O I：

10.2174/138920006776359275

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

Objective: The aim of this study was to evaluate the influence of microsomal incubation conditions on CYP3A4-mediated metabolism of cyclosporine (CsA) within the limits of previous in vitro studies by application of a statistical experimental design. The effect of changes in microsomal incubation conditions (NADPH, Mg2+, Cl-, NADPH-regenerating system and pH) on the formation of the CYP3A4 metabolites AM1 and AM9 from CsA were studied by application of a fractional factorial design. Metabolism was studied in microsomes of transfected human liver epithelial (THLE) cells specifically expressing CYP3A4. Within the conditions tested, a 3-4-fold difference in formation of CsA metabolites was observed. Formation of both AMI and AM9 was favoured by a low Mg2+ concentration (0.5 mM) and no addition of Cl- to the incubation matrix. However, while a high NADPH concentration (1.75 mM) was the single most important factor for the formation of AM1, changes in NADPH concentration between 0.25 and 1.75 mM had no influence on AM9 formation. Formation of the two metabolites also differed in their influence by pH changes, as a change in pH from 7.2 to 7.5 significantly increased the formation of AM9, while fort-nation of AM1 was unaffected by this change. The present study showed that relatively small changes in the incubation matrix had a significant influence on the microsomal CYP3A4-mediated metabolism of CsA. Systematic studies on microsomal incubation conditions could be a key to improve metabolic in vitro-in vivo extrapolations in drug development.

引用

页码：265 / 271

页数：7

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