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Planta Med 2018; 84(05): 320-328
DOI: 10.1055/s-0043-119886
Pharmacokinetic Investigations
Original Papers
Georg Thieme Verlag KG Stuttgart · New York

Comparison of In Vitro Hepatic Scoparone 7-O-Demethylation between Humans and Experimental Animals

Anam Fayyaz
School of Pharmacy, Faculty of Health Sciences, University of Eastern Finland, Kuopio, Finland
,
Seddy Makwinja
School of Pharmacy, Faculty of Health Sciences, University of Eastern Finland, Kuopio, Finland
,
Seppo Auriola
School of Pharmacy, Faculty of Health Sciences, University of Eastern Finland, Kuopio, Finland
,
Hannu Raunio
School of Pharmacy, Faculty of Health Sciences, University of Eastern Finland, Kuopio, Finland
,
Risto O. Juvonen
School of Pharmacy, Faculty of Health Sciences, University of Eastern Finland, Kuopio, Finland
› Author Affiliations
Further Information

Publication History

received 11 April 2017
revised 25 August 2017

accepted 11 September 2017

Publication Date:
26 September 2017 (online)

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Abstract

Scoparone is a natural bioactive compound in Chinese herbal medicines. It has numerous pharmacological actions, including liver protective, hypolipidemic, antitumor, and anti-inflammatory effects. The primary metabolism route of scoparone is O-demethylation to scopoletin or isoscopoletin catalyzed by CYP enzymes. The aims of our study were to identify the human CYP enzymes catalyzing scoparone 7-O-demethylation to scopoletin and to compare this oxidation reaction in liver microsomes among different species. A high throughput fluorescent-based assay method was developed to determine the scoparone 7-O-demethylation to scopoletin rate. The rate was 100 – 400 nmol/(min×g protein) in mouse and rabbit liver microsomes, 10 – 20 nmol/(min×g protein) in pig microsomes, 1 – 3 nmol/(min×g protein) in human and less than 1 nmol/(min×g protein) in rat liver microsomes. Human CYP1A1 (Km 13 µM and Vmax 0.8 min−1), CYP1A2 (Km 48 µM and Vmax 0.3 min−1), and CYP2A13 (Km 10 µM and Vmax 22 min−1) were the most efficient catalysts of the reaction. The CYP2A6 selective inhibitor pilocarpine and an antibody against mouse CYP2A5 inhibited scoparone 7-O-demethylation to scopoletin in rabbit, mouse, and pig liver microsomes, indicating involvement of CYP2A enzymes in the reaction. Hepatic scoparone 7-O-demethylation to scopoletin differed between species both with respect to the rate of reaction and catalyzing enzymes. These species differences need to be taken into account when testing scoparone pharmacokinetics in animals and humans.

Key words

coumarins - in vitro methods - pharmacokinetics - cytochrome P450 - scoparone - TCM
 

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