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Kinetic Characterization of Estradiol Glucuronidation by Liver Microsomes and Expressed UGT Enzymes: The Effects of Organic Solvents

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Abstract

Background and Objective

In vitro glucuronidation of 17β-estradiol (estradiol) is often performed to assess the role of uridine 5′-diphospho-glucuronosyltransferase 1A1 (UGT1A1) in xenobiotic/drug metabolism. The objective of this study was to determine the effects of four commonly used organic solvents [i.e., dimethyl sulfoxide (DMSO), methanol, ethanol, and acetonitrile] on the glucuronidation kinetics of estradiol, which can be glucuronidated at C3 and C17 positions.

Methods

The impacts of organic solvents on estradiol glucuronidation were determined by using expressed UGT enzymes and liver microsomes from both human and animals.

Results

In human liver microsomes (HLM), methanol, ethanol, and acetonitrile significantly altered estradiol glucuronidation kinetics with increased Vmax (up to 2.6-fold) and CLmax (up to 2.8-fold) values. Altered estradiol glucuronidation in HLM was deduced to be attributed to the enhanced metabolic activities of UGT1A1 and UGT2B7, whose activities differ at the two glucuronidation positions. The effects of organic solvents on estradiol glucuronidation were glucuronidation position-, isozyme-, and solvent-specific. Furthermore, both ethanol and acetonitrile have a greater tendency to modify the glucuronidation activity of estradiol in animal liver microsomes.

Conclusion

Organic solvents such as methanol, ethanol, and acetonitrile showed great potential in adjusting the glucuronidation of estradiol. DMSO is the most suitable solvent due to its minimal influence on estradiol glucuronidation. Researchers should be cautious in selecting appropriate solvents to get accurate results when assessing the metabolism of a new chemical entity.

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Author notes

  1. Caimei Wu and Meixue Luo contribute equally to this work.

Authors and Affiliations

  1. Institute of Molecular Rhythm and Metabolism, School of Pharmaceutical Sciences, Guangzhou University of Chinese Medicine, No. 232 Waihuan East Road, Guangzhou Higher Education Mega Center, Panyu District, Guangzhou, 510006, China

    Caimei Wu, Meixue Luo, Dihao **e, Simin Zhong, Jiahao Xu & Danyi Lu

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  1. Caimei Wu
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  2. Meixue Luo
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  3. Dihao **e
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  4. Simin Zhong
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Correspondence to Danyi Lu.

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Authors Contributions

CW, ML, DL: participated in research design; CW, ML, DX, DL: conducted experiments; CW, ML, DX, SZ, JX: performed data analysis; CW, ML, DL: wrote or contributed to the writing of the manuscript; DL: Funding acquisition.

Funding

This work was supported by the Guangdong Basic and Applied Basic Research Foundation (nos. 2023A1515030140 and 2021A1515011256) and the Science and Technology Projects in Guangzhou (no. 202201011284).

Data Availability

The data are available from the corresponding author and shall be shared upon a reasonable request.

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The authors report there are no competing interests to declare.

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Wu, C., Luo, M., **e, D. et al. Kinetic Characterization of Estradiol Glucuronidation by Liver Microsomes and Expressed UGT Enzymes: The Effects of Organic Solvents. Eur J Drug Metab Pharmacokinet 49, 343–353 (2024). https://doi.org/10.1007/s13318-024-00888-2

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