Rates of metabolism of chlorzoxazone, dextromethorphan, 7-ethoxycoumarin, imipramine, quinidine, testosterone and verapamil by fresh and cryopreserved rat liver slices, and some comparisons with microsomes.
In the present study we have investigated the disappearance of chlorzoxazone, dextromethorphan, 7-ethoxycoumarin, imipramine, quinidine, testosterone and verapamil from the medium in which fresh and cryopreserved rat liver slices were incubated. These compounds are all substrates of major isoforms of cytochrome P450 expressed in the liver. The metabolism of five of these compounds in microsomes from rat liver was also examined. Determinations of the concentrations of the compounds were performed employing LC/MS. Intrinsic clearance values (CL(ints)) were calculated on the basis of the concentration-vs.-time curves. No significant differences in the CL(int) values obtained with fresh and cryopreserved rat liver slices were observed for any of the compounds. The highest CL(int) value estimated with liver slices was observed for testosterone and the lowest values were with chlorzoxazone and 7-ethoxycoumarin. The total CL(int) values for 7-ethoxycoumarin and imipramine, calculated using scaling factors, were similar for liver slices and microsomes. In the case of testosterone, this total CL(int) was approximately 3.7-fold lower, whereas for dextromethorphan and quinidine it was 2.5- and 8.5-fold higher, respectively, with liver slices than with microromes. In conclusion, the rate of metabolism of the seven compounds tested with rat liver slices was not affected by cryopreservation. This finding adds further support to the general conclusion that the major activities involved in drug metabolism are not affected by cryopreservation of rat liver slices.
Axelsson
H
H
Research DMPK, AstraZeneca R&D Södertälje, S-151 85, Södertälje, Sweden.
Granhall
C
C
Floby
E
E
Jaksch
Y
Y
Svedling
M
M
Sohlenius-Sternbeck
A-K
AK
D000818:Animals; D000928:Antidepressive Agents / Q000493:pharmacokinetics; D000962:Antimalarials / Q000493:pharmacokinetics; D000996:Antitussive Agents / Q000493:pharmacokinetics; D002121:Calcium Channel Blockers / Q000493:pharmacokinetics; D002470:Cell Survival / Q000187:drug effects; D002753:Chlorzoxazone / Q000493:pharmacokinetics; D004396:Coloring Agents; D003374:Coumarins / Q000493:pharmacokinetics; D015925:Cryopreservation; D003577:Cytochrome P-450 Enzyme System / Q000378:metabolism; D003915:Dextromethorphan / Q000493:pharmacokinetics; D007099:Imipramine / Q000493:pharmacokinetics; D066298:In Vitro Techniques; D008099:Liver / Q000201:enzymology / Q000378:metabolism*; D008297:Male; D008862:Microsomes, Liver / Q000201:enzymology / Q000378:metabolism*; D009125:Muscle Relaxants, Central / Q000493:pharmacokinetics; D010078:Oxazines; D004364:Pharmaceutical Preparations / Q000378:metabolism*; D010599:Pharmacokinetics; D011802:Quinidine / Q000493:pharmacokinetics; D051381:Rats; D017207:Rats, Sprague-Dawley; D013347:Subcellular Fractions / Q000378:metabolism; D013739:Testosterone / Q000493:pharmacokinetics; D014700:Verapamil / Q000493:pharmacokinetics; D014966:Xanthenes
D000928:Antidepressive Agents; D000962:Antimalarials; D000996:Antitussive Agents; D002121:Calcium Channel Blockers; D004396:Coloring Agents; D003374:Coumarins; D009125:Muscle Relaxants, Central; D010078:Oxazines; D004364:Pharmaceutical Preparations; D014966:Xanthenes; C005843:resazurin; C017299:7-ethoxycoumarin; D013739:Testosterone; D003915:Dextromethorphan; D003577:Cytochrome P-450 Enzyme System; D014700:Verapamil; D002753:Chlorzoxazone; D011802:Quinidine; D007099:Imipramine