Abstract
Human arylacetamide deacetylase (AADAC) is a major esterase responsible for the hydrolysis of clinical drugs such as flutamide, phenacetin, and rifampicin. Thus, AADAC is considered to be a relevant enzyme in preclinical drug development, but there is little information about species differences with AADAC. This study investigated the species differences in the tissue distribution and enzyme activities of AADAC. In human, AADAC mRNA was highly expressed in liver and the gastrointestinal tract, followed by bladder. In rat and mouse, AADAC mRNA was expressed in liver at the highest level, followed by the gastrointestinal tract and kidney. The expression levels in rat tissues were approximately 7- and 10-fold lower than those in human and mouse tissues, respectively. To compare the catalytic efficiency of AADAC among three species, each recombinant AADAC was constructed, and enzyme activities were evaluated by normalizing with the expression levels of AADAC. Flutamide and phenacetin hydrolase activities were detected by the recombinant AADAC of all species. In flutamide hydrolysis, liver microsomes of all species showed similar catalytic efficiencies, despite the lower AADAC mRNA expression in rat liver. In phenacetin hydrolysis, rat liver microsomes showed approximately 4- to 6.5-fold lower activity than human and mouse liver microsomes. High rifampicin hydrolase activity was detected only by recombinant human AADAC and human liver and jejunum microsomes. Taken together, the results of this study clarified the species differences in the tissue distribution and enzyme activities of AADAC and facilitate our understanding of species differences in drug hydrolysis.
Footnotes
This study was supported by the Japan Society for the Promotion of Science [Grant-in-Aid for Young Scientists (B) 21790148].
Article, publication date, and citation information can be found at http://dmd.aspetjournals.org.
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The online version of this article (available at http://dmd.aspetjournals.org) contains supplemental material.
ABBREVIATIONS:
- CES
- carboxylesterase
- AADAC
- arylacetamide deacetylase
- FLU-1
- 4-nitro-3-(trifluoromethyl)phenylamine
- RT
- reverse transcription
- PCR
- polymerase chain reaction
- HLM
- human liver microsomes
- HJM
- human jejunum microsomes
- HRM
- human renal microsomes
- PAGE
- polyacrylamide gel electrophoresis
- RRM
- rat renal microsomes
- MJM
- mouse jejunum microsomes
- MRM
- mouse renal microsomes
- MLM
- mouse liver microsomes
- MPM
- mouse pulmonary microsomes.
- Received September 29, 2011.
- Accepted December 29, 2011.
- Copyright © 2012 by The American Society for Pharmacology and Experimental Therapeutics
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