title: In-vivo and in-vitro metabolic clearance of midazolam, a cytochrome P450 3A substrate, by the liver under normal and increased enzyme activity in rats.
abstract: The metabolic clearance of midazolam, a cytochrome P450 (CYP) 3A substrate, by the liver under normal and increased enzyme activity in rats was determined in-vivo and in-vitro to elucidate the reproducibility of the in-vivo hepatic extraction ratio of midazolam from the in-vitro study. The hepatic enzyme activity was modified by pretreating rats with a CYP inducer such as dexamethasone and clotrimazole. The in-vivo hepatic extraction ratio (ERh,obs) of midazolam under a steady-state plasma concentration (approx. 3 nmolmL(-1)) in untreated (control) rats was 0.864. This value increased to 0.984 in dexamethasone-pretreated rats and to 0.964 in clotrimazole-pretreated rats. The in-vitro hepatic intrinsic clearance (CL(int,in-vitro)), expressed as mLmin(-1) (mg microsomal protein)(-1), of midazolam was estimated as Vmax (Km)(-1) by in-vitro metabolism studies using liver microsomes. The CL(int,in-vitro) value was converted to the CL(int,cal) value, expressed as mLmin(-1)kg(-1), by considering the microsomal protein content (g liver)(-1) and the microsomal protein content (g liver)(-1)kg(-1). The estimated CL(int,cal) value was then converted to the ERh value (ER(h,cal)) according to the well-stirred, the parallel-tube and the dispersion models. The ERh(h,cal) values obtained by the parallel-tube model were in good agreement with corresponding in-vivo ERh(h,obs) values. In conclusion, it was demonstrated that high hepatic clearances of midazolam under normal and increased CYP3A activity were reasonably predicted from in-vitro metabolism studies using liver microsomes.
authors.affiliation: Institute of Pharmaceutical Sciences, Hiroshima University School of Medicine, Japan.
authors.forename: F
authors.initials: F
authors.lastname: Higashikawa
authors.forename: T
authors.initials: T
authors.lastname: Murakami
authors.forename: T
authors.initials: T
authors.lastname: Kaneda
authors.forename: M
authors.initials: M
authors.lastname: Takano
mesh_terms: D000284:Administration, Oral; D000818:Animals; D014151:Anti-Anxiety Agents / Q000493:pharmacokinetics*; D001189:Aryl Hydrocarbon Hydroxylases; D003022:Clotrimazole / Q000494:pharmacology; D051544:Cytochrome P-450 CYP3A; D003577:Cytochrome P-450 Enzyme System / Q000378:metabolism*; D003907:Dexamethasone / Q000494:pharmacology; D004789:Enzyme Activation / Q000187:drug effects; D066298:In Vitro Techniques; D007413:Intestinal Mucosa / Q000378:metabolism; D008099:Liver / Q000201:enzymology / Q000378:metabolism*; D008297:Male; D008657:Metabolic Clearance Rate; D008862:Microsomes, Liver / Q000378:metabolism; D008874:Midazolam / Q000378:metabolism / Q000493:pharmacokinetics*; D010089:Oxidoreductases, N-Demethylating / Q000378:metabolism*; D051381:Rats; D017207:Rats, Sprague-Dawley; D013379:Substrate Specificity; D013997:Time Factors
chemical_list: D014151:Anti-Anxiety Agents; D003907:Dexamethasone; D003577:Cytochrome P-450 Enzyme System; D001189:Aryl Hydrocarbon Hydroxylases; D051544:Cytochrome P-450 CYP3A; D010089:Oxidoreductases, N-Demethylating; D003022:Clotrimazole; D008874:Midazolam