Non-invasive method to detect induction of CYP3A4 in chimeric mice with a humanized liver.
Chimeric mice with a humanized liver have been previously established by the transplantation of human hepatocytes to urokinase-type plasminogen activator/severe combined immunodeficiency mice. A non-invasive method to detect the induction of cytochrome P450 (CYP) 3A4 was evaluated in chimeric mice with a humanized liver. Dexamethasone (DEX) was used as a probe drug to detect induction; and rifampicin was used as a model drug to induce CYP3A4. Before and after rifampicin treatment (50 mg kg(-1), intraperitoneal injection once a day for 4 days) in the chimeric mice, DEX was subcutaneously injected and the urinary excretion of 6beta-hydroxydexamethason (6betaOHD) and DEX was determined. The metabolic ratio (6betaOHD/DEX) significantly increased after rifampicin treatment. Livers from the control and rifampicin-treated chimeric mice were stained immunohistolochemically with antibodies against CYP3A4 and CYP3A5. CYP3A4 and CYP3A5 were detected in the area of humanized liver, but staining was intense for CYP3A4 and very weak for CYP3A5. Only the staining of CYP3A4 was increased after rifampicin treatment. Formation of 6betaOHD by human liver microsomes was higher than that formed by mouse liver microsomes. Metabolite formation was catalysed by both CYP3A4 and CYP3A5 and the intrinsic clearance (V(max)/K(m)) by CYP3A4 was found to be 50-fold higher than that of CYP3A5. The results of the present study indicate that estimation of the changes of the urinary metabolic ratio (6betaOHD/DEX) in the chimeric mice with a humanized liver is a very useful tool for detecting the induction of CYP3A4 by a non-invasive method.
Emoto
C
C
Pharmacokinetics Dynamics Metabolism, Pfizer Global Research & Development, Nagoya Laboratories, Pfizer Japan, Inc., Aichi, Japan.
Yamato
Y
Y
Sato
Y
Y
Ohshita
H
H
Katoh
M
M
Tateno
C
C
Yokoi
T
T
Yoshizato
K
K
Iwasaki
K
K
D000818:Animals; D001681:Biological Assay / Q000379:methods*; D002678:Chimera / Q000378:metabolism*; D002845:Chromatography; D051544:Cytochrome P-450 CYP3A / Q000096:biosynthesis*; D003907:Dexamethasone / Q000008:administration & dosage / Q000031:analogs & derivatives / Q000494:pharmacology / Q000652:urine; D004790:Enzyme Induction / Q000187:drug effects; D006801:Humans; D007150:Immunohistochemistry; D007700:Kinetics; D008099:Liver / Q000187:drug effects / Q000201:enzymology*; D051379:Mice; D008862:Microsomes, Liver / Q000187:drug effects / Q000201:enzymology; D011994:Recombinant Proteins / Q000378:metabolism; D012293:Rifampin / Q000494:pharmacology; D013379:Substrate Specificity / Q000187:drug effects
D011994:Recombinant Proteins; C051956:6-hydroxydexamethasone; D003907:Dexamethasone; D051544:Cytochrome P-450 CYP3A; C510163:CYP3A4 protein, human; D012293:Rifampin