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Use of Isoform-Specific UGT Metabolism to Determine and Describe Rates and Profiles of Glucuronidation of Wogonin and Oroxylin A by Human Liver and Intestinal Microsomes

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ABSTRACT

Purposes

Glucuronidation via UDP-glucuronosyltransferases (or UGTs) is a major metabolic pathway. The purposes of this study are to determine the UGT-isoform-specific metabolic fingerprint (or GSMF) of wogonin and oroxylin A, and to use isoform-specific metabolism rates and kinetics to determine and describe their glucuronidation behaviors in tissue microsomes.

Methods

In vitro glucuronidation rates and profiles were measured using expressed UGTs and human intestinal and liver microsomes.

Results

GSMF experiments indicated that both flavonoids were metabolized mainly by UGT1As, with major contributions from UGT1A3 and UGT1A7-1A10. Isoform-specific metabolism showed that kinetic profiles obtained using expressed UGT1A3 and UGT1A7-1A10 could fit to known kinetic models. Glucuronidation of both flavonoids in human intestinal and liver microsomes followed simple Michaelis-Menten kinetics. A comparison of the kinetic parameters and profiles suggests that UGT1A9 is likely the main isoform responsible for liver metabolism. In contrast, a combination of UGT1As with a major contribution from UGT1A10 contributed to their intestinal metabolism. Correlation studies clearly showed that UGT isoform-specific metabolism could describe their metabolism rates and profiles in human liver and intestinal microsomes.

Conclusion

GSMF and isoform-specific metabolism profiles can determine and describe glucuronidation rates and profiles in human tissue microsomes.

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Abbreviations

AIC:

Akaike’s information criterion

DHF:

di-hydroxyl flavone

DHFG:

di-hydroxyl flavone glucuronide

GSMF:

UGT-isoform specific metabolic fingerprint

HIM:

human intestinal microsomes

HLM:

human liver microsomes

MAICE:

minimum AIC estimation

UDPGA:

Uridine diphosphate glucuronic acid

UGTs:

UDP-glucuronosyltransferases

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ACKNOWLEDGEMENTS

This work was mainly supported by the Ministry of Science and Technology of the People’s Republic of China Grants 2006BAT11B08-4, and the Grant of Science and Technology of Guangzhou 2006Z1-E6021, both to ZL. MH was also supported by NIH GM070737. The authors would like to thank Dr. Vincent Tam of University of Houston College of Pharmacy for his help in rendering the program to analyze the kinetics of glucuronidation by UGT 1A3. The authors would like to acknowledge Ms. Ling Ye for her technical assistance in the measurement of metabolites using LC-MS/MS. Additional technical assistance by Ms. Jie Zhao in the determination of conversion factor is also acknowledged here.

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Authors and Affiliations

  1. Department of Pharmaceutics, School of Pharmaceutical Sciences, Southern Medical University, 1838 North Guangzhou Avenue, Guangzhou, China, 510515

    Qiong Zhou, Zhijie Zheng, Bijun Xia, Lan Tang, Chang Lv, Wei Liu, Zhongqiu Liu & Ming Hu

  2. Department of Pharmacological and Pharmaceutical Sciences College of Pharmacy, University of Houston, 1441 Moursund Street, Houston, TEXAS, 77030, USA

    Ming Hu

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  1. Qiong Zhou
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  4. Lan Tang
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Correspondence to Zhongqiu Liu or Ming Hu.

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Zhou, Q., Zheng, Z., Xia, B. et al. Use of Isoform-Specific UGT Metabolism to Determine and Describe Rates and Profiles of Glucuronidation of Wogonin and Oroxylin A by Human Liver and Intestinal Microsomes. Pharm Res 27, 1568–1583 (2010). https://doi.org/10.1007/s11095-010-0148-0

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  • DOI: https://doi.org/10.1007/s11095-010-0148-0

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