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image of `3-Hydroxy-6H,7H-chromeno [3,4-c]chromene-6,7-dione as a Substrate for Studying Glucuronidation, Sulfonation and Lactone Hydrolysis

Abstract

Introduction/Objective

Glucuronidation and sulfonation enzymes conjugate compounds containing a hydroxyl group, while paraoxonase enzymes hydrolyze lactone-containing compounds. 3-Hydroxy-6H,7H-chromeno [3,4-c]chromene-6,7-dione (3-hydroxy-V-coumarin) contains both a hydroxyl substituent and two lactone substructures and is strongly fluorescent. This study evaluates glucuronidation, sulfonation and hydrolysis of 3-hydroxy-V-coumarin reactions, which abolish its fluorescence.

Methods

Glucuronide, sulfate, and hydrolyzed product formation were confirmed using accurate LC-MS. Fluorescence-based multi-well plate assays were established to determine rates of glucuronidation, sulfonation and hydrolysis of 3-hydroxy-V-coumarin.

Results

A decrease in fluorescence correlated with the formation of conjugation or hydrolyzed metabolites of the reactions was observed. 3-Hydroxy-V-coumarin was glucuronidated by human UGT1A1, 1A3, 1A4, 1A7, 1A8, 1A9, 1A10, 2A1, 2B4 and 2B7 and by dog UGT1A1, 1A2 and 1A11, and sulfonated by human SULT1A1, 1A2 and 1E1. The results indicated that paraoxonase 2 hydrolyzed lactone of 3-hydroxy-V-coumarin. 3-Hydroxy-V-coumarin interacted a hydrogen bond with serine 221 and hydrophobic interaction with phenyls 71 and 291 of paraoxonase 2. The hydrolysis rate of 3-hydroxy-V-coumarin was determined in ten rat tissues. In human liver cytosol, the hydrolysis was slower than in rat, mouse, dog, pig, rabbit, and sheep liver cytosol.

Conclusion

3-hydroxy-V-coumarin is a new model substrate for studying glucuronidation, sulfonation and lactone hydrolysis reactions

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2025-04-25
2025-08-17

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`3-Hydroxy-6H,7H-chromeno [3,4-c]chromene-6,7-dione as a Substrate for Studying Glucuronidation, Sulfonation and Lactone Hydrolysis
Bentham Science Publishers ; https://doi.org/10.2174/0118723128351379250312063255
/content/journals/dmbl/10.2174/0118723128351379250312063255
/content/journals/dmbl/10.2174/0118723128351379250312063255
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  • Article Type:     Research Article
Keywords: sulfonation ; paraoxonase ; Hydrolysis ; lactone ; fluorescence ; glucuronidation

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