Molecular docking and enzyme kinetic studies of dihydrotanshinone on metabolism of a model CYP2D6 probe substrate in human liver microsomes

Xuelin Zhou, Yan Wang, Penelope M Y Or, David C C Wan, Yiu Wa Kwan, John H K Yeung

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10 Scopus citations

Abstract

The effects of Danshen and its active components (tanshinone I, tanshinone IIA, dihydrotanshinone and cryptotanshinone) on CYP2D6 activity was investigated by measuring the metabolism of a model CYP2D6 probe substrate, dextromethorphan to dextrorphan in human pooled liver microsomes. The ethanolic extract of crude Danshen (6.25-100 μg/ml) decreased dextromethorphan O-demethylation in vitro (IC 50 = 23.3 μg/ml) and the water extract of crude Danshen (0.0625-1 mg/ml) showed no inhibition. A commercially available Danshen pill (31.25-500 μg/ml) also decreased CYP2D6 activity (IC 50 = 265.8 μg/ml). Among the tanshinones, only dihydrotanshinone significantly inhibited CYP2D6 activity (IC 50 = 35.4 μM), compared to quinidine, a specific CYP2D6 inhibitor (IC 50 = 0.9 μM). Crytotanshinone, tanshinone I and tanshinone IIA produced weak inhibition, with IC 20 of 40.8 μM, 16.5 μM and 61.4 μM, respectively. Water soluble components such as salvianolic acid B and danshensu did not affect CYP2D6-mediated metabolism. Enzyme kinetics studies showed that inhibition of CYP2D6 activity by the ethanolic extract of crude Danshen and dihydrotanshinone was concentration-dependent, with K i values of 4.23 μg/ml and 2.53 μM, respectively, compared to quinidine, K i = 0.41 μM. Molecular docking study confirmed that dihydrotanshinone and tanshinone I interacted with the Phe120 amino acid residue in the active cavity of CYP2D6 through Pi-Pi interaction, but did not interact with Glu216 and Asp301, the key residues for substrate binding. The logarithm of free binding energy of dihydrotanshinone (-7.6 kcal/mol) to Phe120 was comparable to quinidine (-7.0 kcal/mol) but greater than tanshinone I (-5.4 kcal/mol), indicating dihydrotanshinone has similar affinity to quinidine in binding to the catalytic site on CYP2D6.

Original languageEnglish (US)
Pages (from-to)648-657
Number of pages10
JournalPhytomedicine
Volume19
Issue number7
DOIs
StatePublished - May 15 2012

Bibliographical note

Funding Information:
The authors gratefully acknowledge the supply of Danshen extract from Winsor Health Products Ltd. (Hong Kong). The work described in this paper was partly supported by a grant from the Research Grants Council of the Hong Kong Special Administrative Region, China (Project no. CUHK4517/06M).

Copyright:
Copyright 2012 Elsevier B.V., All rights reserved.

Keywords

  • CYP2D6 activity
  • Danshen (Salvia miltiorrhiza)
  • Dextromethorphan metabolism
  • Dihydrotanshinone
  • Human liver microsomes
  • Molecular docking

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