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Drug Metabolism and Disposition Fast Forward
First published on May 12, 2008; DOI: 10.1124/dmd.108.020560

0090-9556/08/3608-1562-1569$20.00
DMD 36:1562-1569, 2008

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UDP-Glucuronosyltransferase 1A6 Is the Major Isozyme Responsible for Protocatechuic Aldehyde Glucuronidation in Human Liver Microsomes

Hui-Xin Liu, Yong Liu, Jiang-Wei Zhang, Wei Li, Hong-Tao Liu, and Ling Yang

Laboratory of Pharmaceutical Resource Discovery (H.-X.L., Y.L., J.-W.Z., W.L., H.-T.L., L.Y.), Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian, China; and Graduate University of Chinese Academy of Sciences, Beijing, China (H.-X.L., J.-W.Z., W.L., H.-T.L.)

Glucuronidation is an important pathway in the metabolism of protocatechuic aldehyde (3,4-dihydroxybenzaldehyde, PAL). However, the metabolites and primary UDP-glucuronosyltransferase (UGT) isozymes responsible for PAL glucuronidation remain to be determined in human. Here, we characterized PAL glucuronidation by human liver microsomes (HLMs), human intestine microsomes (HIMs), and 12 recombinant UGT (rUGT) isozymes to identify what kinds of metabolites are present and which human UGT isozymes are involved. Two metabolites (M-1 and M-2) were detected in reactions catalyzed by HLMs, HIMs, rUGT1A6, and rUGT1A9 and were identified as monoglucuronides by liquid chromatography-mass spectrometry. A kinetic study showed that PAL glucuronidation by rUGT1A6, rUGT1A9, HIMs, and HLMs followed Michaelis-Menten kinetics. The Km values of HLMs, HIMs, rUGT1A6, and rUGT1A9 for PAL glucuronidation were as follows: 432.7 ± 24.5, 626.9 ± 49.2, 367.5 ± 25.1, and 379.9 ± 42.5 µM for M-1 and 336.7 ± 15.3, 494.3 ± 48.7, 211.4 ± 13.4, and 238.5 ± 26.2 µM for M-2, respectively. The PAL glucuronidation activity was significantly correlated with UGT1A6 activity rather than with UGT1A9 activity from 15 individual HLMs. A chemical inhibition study showed that the IC50 for phenylbutazone inhibition of PAL glucuronidation was similar in HLMs (61.9 ± 7.9 µM) compared with rUGT1A6 (45.3 ± 7.7 µM). In contrast, androsterone inhibited rUGT1A9-catalyzed and HLM-catalyzed PAL glucuronidation with IC50 values of 27.1 ± 3.8 and >500 µM, respectively. In combination, we identified UGT1A6 as the major isozyme responsible for PAL glucuronidation in HLMs.

Address correspondence to: Dr. Ling Yang, Laboratory of Pharmaceutical Resource Discovery, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, No. 457 Zhongshan Rd., Dalian 116023 China. E-mail: yling{at}dicp.ac.cn






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