title: Prediction of metabolic clearance of diclofenac in adjuvant-induced arthritis rats using a substrate depletion assay.
abstract: 1. The purpose of this study was to evaluate drug clearance measured by the metabolic intrinsic clearance (CL(int)) in a substrate depletion assay in comparison with the in vivo clearance (CL(tot)) observed in adjuvant-induced arthritis (AA) rats. 2. After intravenous administration of diclofenac as a model drug, CL(tot) was 2.8-fold higher in AA rats than in control rats. In two different substrate depletion assays with liver microsomes for glucuronidation and hydroxylation, the CL(int) values for glucuronidation was significantly decreased in AA rats to 60% of the value in control rats, whereas the CL(int) values for hydroxylation were similar. The unbound fraction of diclofenac in plasma (f(u, plasma)) was significantly higher (2.8-fold) in AA rats than in control rats. 3. Hepatic clearance predicted from the CL(int) values for both biotransformation pathways and f(u, plasma) was higher in AA rats than in control rats, with good consistency between predicted and observed values. The same results were obtained for experiments using hepatocytes. 4. The plasma protein-binding activities, rather than metabolic clearance, in both types of rats would be a determining factor in the pharmacokinetic behaviour differences between control and AA rats. 5. In summary, substrate depletion assays with liver microsomes and hepatocytes in combination with protein binding assessment can help to predict changes in pharmacokinetics under AA conditions.
authors.affiliation: Department of Pharmacy, School of Pharmacy, Kinki University, Osaka, Japan.
authors.forename: S
authors.initials: S
authors.lastname: Uno
authors.forename: A
authors.initials: A
authors.lastname: Fujii
authors.forename: H
authors.initials: H
authors.lastname: Komura
authors.forename: A
authors.initials: A
authors.lastname: Kawase
authors.forename: M
authors.initials: M
authors.lastname: Iwaki
mesh_terms: D000818:Animals; D000894:Anti-Inflammatory Agents, Non-Steroidal / Q000493:pharmacokinetics*; D001169:Arthritis, Experimental / Q000188:drug therapy / Q000378:metabolism*; D001711:Biotransformation; D000072467:Cytochrome P450 Family 2; D004008:Diclofenac / Q000493:pharmacokinetics*; D005260:Female; D020719:Glucuronides / Q000378:metabolism; D006900:Hydroxylation; D008657:Metabolic Clearance Rate; D008862:Microsomes, Liver / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D013255:Steroid 21-Hydroxylase / Q000378:metabolism
chemical_list: D000894:Anti-Inflammatory Agents, Non-Steroidal; D020719:Glucuronides; D004008:Diclofenac; C089877:Cyp2c6v1 protein, rat; D000072467:Cytochrome P450 Family 2; D013255:Steroid 21-Hydroxylase