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In vitro identification of cytochrome P450 isoforms responsible for the metabolism of 1-hydroxyl-2,3,5-trimethoxy-xanthone purified from Halenia elliptica D. Don.
Chem Biol Interact. 2014 Mar 05; 210:12-9.CB

Abstract

1-Hydroxyl-2,3,5-trimethoxyxanthone (HM-1) is one of the main constituents extracted from Halenia elliptica D. Don, which is a traditionally used Tibetan medicinal plant. The aim of this study was to illustrate the proposed metabolic pathways of HM-1 and identify which cytochrome P450 (CYP450) isoforms involved in its metabolism by using pooled human liver microsomes (HLMs) and recombinant CYP450 isoforms with selective chemical inhibitors. Metabolites were identified by high performance liquid chromatography coupled to ion trap time-of-flight mass spectrometry (LCMS(n)-ESI-IT-TOF) and nuclear magnetic resonance spectroscopy (hydrogen-1 NMR and carbon-13 NMR). Three metabolites (M1-M3) were identified, which demonstrated that demethylation and hydroxylation were the major Phase I metabolic reactions for HM-1 in HLMs. The structure of another metabolite (M4) was still unclear. The enzymatic kinetics of M1 (K(m)=23.19±14.20 μM) and M2 (Km=32.06±17.09 μM) exhibited substrate inhibition; whereas, the formation of M3 (K(m)=5.73±0.70 μM) and M4 (K(m)=16.43±5.12 μM) displayed Michaelis-Menten kinetics. The intrinsic clearance (V(max)/K(m)) of M3 was highest among these metabolites, suggesting that M3 was the major metabolite of HM-1. Moreover, CYP3A4 and CYP2C8 were the primary CYP450 isoform responsible for the metabolism of HM-1. CYP1A2, CYP2A6, CYP2B6, CYP2C9 and CYP2C19 were also involved in HM-1 metabolism, especially in the formation of M3. This study finally provides evidence of substrate inhibition and metabolism-based drug-drug interaction for the medicinal preparations containing HM-1 used in clinic.

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Authors+Show Affiliations

State Key Laboratory of Bioactive Substances and Functions of Natural Medicines, Institute of Materia Medica, Chinese Academy of Medical Sciences, Beijing 100050, China.

School of Biomedical Sciences, Faculty of Medicine, The Chinese University of Hong Kong, Shatin, N.T., Hong Kong, China.

State Key Laboratory of Bioactive Substances and Functions of Natural Medicines, Institute of Materia Medica, Chinese Academy of Medical Sciences, Beijing 100050, China.

School of Biomedical Sciences, Faculty of Medicine, The Chinese University of Hong Kong, Shatin, N.T., Hong Kong, China.

School of Biomedical Sciences, Faculty of Medicine, The Chinese University of Hong Kong, Shatin, N.T., Hong Kong, China.

State Key Laboratory of Bioactive Substances and Functions of Natural Medicines, Institute of Materia Medica, Chinese Academy of Medical Sciences, Beijing 100050, China.

State Key Laboratory of Bioactive Substances and Functions of Natural Medicines, Institute of Materia Medica, Chinese Academy of Medical Sciences, Beijing 100050, China.

State Key Laboratory of Bioactive Substances and Functions of Natural Medicines, Institute of Materia Medica, Chinese Academy of Medical Sciences, Beijing 100050, China.

State Key Laboratory of Bioactive Substances and Functions of Natural Medicines, Institute of Materia Medica, Chinese Academy of Medical Sciences, Beijing 100050, China.

State Key Laboratory of Bioactive Substances and Functions of Natural Medicines, Institute of Materia Medica, Chinese Academy of Medical Sciences, Beijing 100050, China.

Department of Medicinal Chemistry & Pharmacognosy (MC 781), UIC College of Pharmacy, Chicago, IL 60612-7231, USA.

School of Biomedical Sciences, Faculty of Medicine, The Chinese University of Hong Kong, Shatin, N.T., Hong Kong, China.

State Key Laboratory of Bioactive Substances and Functions of Natural Medicines, Institute of Materia Medica, Chinese Academy of Medical Sciences, Beijing 100050, China. Electronic address: wangyan@imm.ac.cn.

MeSH

Chromatography, High Pressure LiquidCytochrome P-450 Enzyme SystemGentianaceaeHumansKineticsMagnetic Resonance SpectroscopyMicrosomes, LiverMolecular StructurePlants, MedicinalProtein IsoformsTibetXanthones

Pub Type(s)

Journal Article

Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

24380714

Citation

Feng, Ru, et al. "In Vitro Identification of Cytochrome P450 Isoforms Responsible for the Metabolism of 1-hydroxyl-2,3,5-trimethoxy-xanthone Purified From Halenia Elliptica D. Don." Chemico-biological Interactions, vol. 210, 2014, pp. 12-9.

Feng R, Zhou X, Tan XS, et al. In vitro identification of cytochrome P450 isoforms responsible for the metabolism of 1-hydroxyl-2,3,5-trimethoxy-xanthone purified from Halenia elliptica D. Don. Chem Biol Interact. 2014;210:12-9.

Feng, R., Zhou, X., Tan, X. S., Or, P. M., Hu, T., Fu, J., Ma, J. Y., Huang, M., He, C. Y., Shi, J. G., Che, C. T., Yeung, J. H., & Wang, Y. (2014). In vitro identification of cytochrome P450 isoforms responsible for the metabolism of 1-hydroxyl-2,3,5-trimethoxy-xanthone purified from Halenia elliptica D. Don. Chemico-biological Interactions, 210, 12-9. https://doi.org/10.1016/j.cbi.2013.12.008

Feng R, et al. In Vitro Identification of Cytochrome P450 Isoforms Responsible for the Metabolism of 1-hydroxyl-2,3,5-trimethoxy-xanthone Purified From Halenia Elliptica D. Don. Chem Biol Interact. 2014 Mar 5;210:12-9. PubMed PMID: 24380714.

* Article titles in AMA citation format should be in sentence-case

TY - JOUR T1 - In vitro identification of cytochrome P450 isoforms responsible for the metabolism of 1-hydroxyl-2,3,5-trimethoxy-xanthone purified from Halenia elliptica D. Don. AU - Feng,Ru, AU - Zhou,Xuelin, AU - Tan,Xiang-Shan, AU - Or,Penelope M Y, AU - Hu,Tao, AU - Fu,Jie, AU - Ma,Jing-Yi, AU - Huang,Min, AU - He,Chi-Yu, AU - Shi,Jian-Gong, AU - Che,Chun-Tao, AU - Yeung,John H K, AU - Wang,Yan, Y1 - 2013/12/28/ PY - 2013/10/08/received PY - 2013/12/11/revised PY - 2013/12/17/accepted PY - 2014/1/2/entrez PY - 2014/1/2/pubmed PY - 2014/4/9/medline KW - Cytochrome P450 KW - Halenia elliptica KW - Human liver microsomes KW - LCMS(n)-IT-TOF KW - Xanthone SP - 12 EP - 9 JF - Chemico-biological interactions JO - Chem Biol Interact VL - 210 N2 - 1-Hydroxyl-2,3,5-trimethoxyxanthone (HM-1) is one of the main constituents extracted from Halenia elliptica D. Don, which is a traditionally used Tibetan medicinal plant. The aim of this study was to illustrate the proposed metabolic pathways of HM-1 and identify which cytochrome P450 (CYP450) isoforms involved in its metabolism by using pooled human liver microsomes (HLMs) and recombinant CYP450 isoforms with selective chemical inhibitors. Metabolites were identified by high performance liquid chromatography coupled to ion trap time-of-flight mass spectrometry (LCMS(n)-ESI-IT-TOF) and nuclear magnetic resonance spectroscopy (hydrogen-1 NMR and carbon-13 NMR). Three metabolites (M1-M3) were identified, which demonstrated that demethylation and hydroxylation were the major Phase I metabolic reactions for HM-1 in HLMs. The structure of another metabolite (M4) was still unclear. The enzymatic kinetics of M1 (K(m)=23.19±14.20 μM) and M2 (Km=32.06±17.09 μM) exhibited substrate inhibition; whereas, the formation of M3 (K(m)=5.73±0.70 μM) and M4 (K(m)=16.43±5.12 μM) displayed Michaelis-Menten kinetics. The intrinsic clearance (V(max)/K(m)) of M3 was highest among these metabolites, suggesting that M3 was the major metabolite of HM-1. Moreover, CYP3A4 and CYP2C8 were the primary CYP450 isoform responsible for the metabolism of HM-1. CYP1A2, CYP2A6, CYP2B6, CYP2C9 and CYP2C19 were also involved in HM-1 metabolism, especially in the formation of M3. This study finally provides evidence of substrate inhibition and metabolism-based drug-drug interaction for the medicinal preparations containing HM-1 used in clinic. SN - 1872-7786 UR - https://www.unboundmedicine.com/medline/citation/24380714/In_vitro_identification_of_cytochrome_P450_isoforms_responsible_for_the_metabolism_of_1_hydroxyl_235_trimethoxy_xanthone_purified_from_Halenia_elliptica_D__Don_ DB - PRIME DP - Unbound Medicine ER -