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An Investigation on Glucuronidation Metabolite Identification, Isozyme Contribution, and Species Differences of GL-V9 In Vitro and In Vivo

GL-V9 is a prominent derivative of wogonin with a wide therapeutic spectrum and potent anti-tumor activity. The metabolism characteristics of GL-V9 remain unclear. This study aimed to clarify the metabolic pathway of GL-V9 and investigate the generation of its glucuronidation metabolites in vitro an...

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Autores principales: Xing, Han, Kong, Dexuan, Ning, Chen, Kong, Ying, Ren, Chang, Cheng, Yujie, Cai, Hui, Wang, Jubo, Zhao, Di, Li, Ning, Chen, Xijing, Li, Zhiyu, Lu, Yang
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6515479/
https://www.ncbi.nlm.nih.gov/pubmed/31013570
http://dx.doi.org/10.3390/molecules24081576
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author Xing, Han
Kong, Dexuan
Ning, Chen
Kong, Ying
Ren, Chang
Cheng, Yujie
Cai, Hui
Wang, Jubo
Zhao, Di
Li, Ning
Chen, Xijing
Li, Zhiyu
Lu, Yang
author_facet Xing, Han
Kong, Dexuan
Ning, Chen
Kong, Ying
Ren, Chang
Cheng, Yujie
Cai, Hui
Wang, Jubo
Zhao, Di
Li, Ning
Chen, Xijing
Li, Zhiyu
Lu, Yang
author_sort Xing, Han
collection PubMed
description GL-V9 is a prominent derivative of wogonin with a wide therapeutic spectrum and potent anti-tumor activity. The metabolism characteristics of GL-V9 remain unclear. This study aimed to clarify the metabolic pathway of GL-V9 and investigate the generation of its glucuronidation metabolites in vitro and in vivo. HPLC-UV-TripleTOF was used to identify metabolites. The main metabolite that we found was chemically synthesized and the synthetic metabolite was utilized as standard substance for the subsequent metabolism studies of GL-V9, including enzyme kinetics in liver microsomes of five different species and reaction phenotyping metabolism using 12 recombinant human UDP-glucuronosyltransferase (UGT) isoforms. Results indicated that the glucuronidation reaction occurred at C5-OH group, and 5-O-glucuronide GL-V9 is the only glucuronide metabolite and major phase II metabolite of GL-V9. Among 12 recombinant human UGTs, rUGT1A9 showed the strongest catalytic capacity for the glucuronidation reaction of GL-V9. rUGT1A7 and rUGT1A8 were also involved in the glucuronidation metabolism. K(m) of rUGT1A7-1A9 was 3.25 ± 0.29, 13.92 ± 1.05, and 4.72 ± 0.28 μM, respectively. In conclusion, 5-O-glucuronide GL-V9 is the dominant phase II metabolite of GL-V9 in vivo and in vitro, whose formation rate and efficiency are closely related to isoform-specific metabolism profiles and the distribution of UGTs in different tissues of different species.
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spelling pubmed-65154792019-05-30 An Investigation on Glucuronidation Metabolite Identification, Isozyme Contribution, and Species Differences of GL-V9 In Vitro and In Vivo Xing, Han Kong, Dexuan Ning, Chen Kong, Ying Ren, Chang Cheng, Yujie Cai, Hui Wang, Jubo Zhao, Di Li, Ning Chen, Xijing Li, Zhiyu Lu, Yang Molecules Article GL-V9 is a prominent derivative of wogonin with a wide therapeutic spectrum and potent anti-tumor activity. The metabolism characteristics of GL-V9 remain unclear. This study aimed to clarify the metabolic pathway of GL-V9 and investigate the generation of its glucuronidation metabolites in vitro and in vivo. HPLC-UV-TripleTOF was used to identify metabolites. The main metabolite that we found was chemically synthesized and the synthetic metabolite was utilized as standard substance for the subsequent metabolism studies of GL-V9, including enzyme kinetics in liver microsomes of five different species and reaction phenotyping metabolism using 12 recombinant human UDP-glucuronosyltransferase (UGT) isoforms. Results indicated that the glucuronidation reaction occurred at C5-OH group, and 5-O-glucuronide GL-V9 is the only glucuronide metabolite and major phase II metabolite of GL-V9. Among 12 recombinant human UGTs, rUGT1A9 showed the strongest catalytic capacity for the glucuronidation reaction of GL-V9. rUGT1A7 and rUGT1A8 were also involved in the glucuronidation metabolism. K(m) of rUGT1A7-1A9 was 3.25 ± 0.29, 13.92 ± 1.05, and 4.72 ± 0.28 μM, respectively. In conclusion, 5-O-glucuronide GL-V9 is the dominant phase II metabolite of GL-V9 in vivo and in vitro, whose formation rate and efficiency are closely related to isoform-specific metabolism profiles and the distribution of UGTs in different tissues of different species. MDPI 2019-04-22 /pmc/articles/PMC6515479/ /pubmed/31013570 http://dx.doi.org/10.3390/molecules24081576 Text en © 2019 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Xing, Han
Kong, Dexuan
Ning, Chen
Kong, Ying
Ren, Chang
Cheng, Yujie
Cai, Hui
Wang, Jubo
Zhao, Di
Li, Ning
Chen, Xijing
Li, Zhiyu
Lu, Yang
An Investigation on Glucuronidation Metabolite Identification, Isozyme Contribution, and Species Differences of GL-V9 In Vitro and In Vivo
title An Investigation on Glucuronidation Metabolite Identification, Isozyme Contribution, and Species Differences of GL-V9 In Vitro and In Vivo
title_full An Investigation on Glucuronidation Metabolite Identification, Isozyme Contribution, and Species Differences of GL-V9 In Vitro and In Vivo
title_fullStr An Investigation on Glucuronidation Metabolite Identification, Isozyme Contribution, and Species Differences of GL-V9 In Vitro and In Vivo
title_full_unstemmed An Investigation on Glucuronidation Metabolite Identification, Isozyme Contribution, and Species Differences of GL-V9 In Vitro and In Vivo
title_short An Investigation on Glucuronidation Metabolite Identification, Isozyme Contribution, and Species Differences of GL-V9 In Vitro and In Vivo
title_sort investigation on glucuronidation metabolite identification, isozyme contribution, and species differences of gl-v9 in vitro and in vivo
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6515479/
https://www.ncbi.nlm.nih.gov/pubmed/31013570
http://dx.doi.org/10.3390/molecules24081576
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