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. 2020 Sep 24;12(10):916.
doi: 10.3390/pharmaceutics12100916.

Quasi-Irreversible Inhibition of CYP2D6 by Berberine

Ha Gyeong Kim  1 , Han Sol Lee  1 , Jang Su Jeon  1 , Young Jae Choi  1 , Yeon Jung Choi  1 , So-Yeol Yoo  1 , Eun-Yeong Kim  2 , Kiho Lee  2 , InWha Park  1   3 , MinKyun Na  1 , Han-Jin Park  4 , Seung-Woo Cho  5 , Jong-Hoon Kim  6 , Jae-Young Lee  1 , Sang Kyum Kim  1
Affiliations

Quasi-Irreversible Inhibition of CYP2D6 by Berberine

Ha Gyeong Kim et al. Pharmaceutics. .

Abstract

In our previous study, Hwang-Ryun-Hae-Dok-Tang, which contains berberine (BBR) as a main active ingredient, inhibited cytochrome P450 (CYP) 2D6 in a quasi-irreversible manner. However, no information is available on the detailed mechanism of BBR-induced CYP2D6 inhibition. Thus, the present study aimed to characterize the inhibition mode and kinetics of BBR and its analogues against CYP2D6 using pooled human liver microsomes (HLM). BBR exhibited selective quasi-irreversible inhibition of CYP2D6 with inactivation rate constant (kinact) of 0.025 min-1, inhibition constant (KI) of 4.29 µM, and kinact/KI of 5.83 mL/min/µmol. In pooled HLM, BBR was metabolized to thalifendine (TFD), demethyleneberberine (DMB), M1 (proposed as demethylene-TFD), and to a lesser extent berberrubine (BRB), showing moderate metabolic stability with a half-life of 35.4 min and a microsomal intrinsic clearance of 7.82 µL/min/mg protein. However, unlike BBR, those metabolites (i.e., TFD, DMB, and BRB) were neither selective nor potent inhibitors of CYP2D6, based on comparison of half-maximal inhibitory concentration (IC50). Notably, TFD, but not DMB, exhibited metabolism-dependent CYP2D6 inhibition as in the case of BBR, which suggests that methylenedioxybenzene moiety of BBR may play a critical role in the quasi-irreversible inhibition. Moreover, the metabolic clearance of nebivolol (β-blocker; CYP2D6 substrate) was reduced in the presence of BBR. The present results warrant further evaluation of BBR-drug interactions in clinical situations.

Keywords: berberine; cytochrome P450; drug metabolism; herb-drug interaction; metabolite identification.

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Conflict of interest statement

The authors declare no conflict of interest. The funders had no role in the design of the study; in the collection, analyses, or interpretation of data; in the writing of the manuscript, or in the decision to publish the results.

Figures

Figure 1
Figure 1
Effects of berberine and tetrahydroberberine on CYP1A2 (A), CYP2A6 (B), CYP2B6 (C), CYP2C8 (D), CYP2C9 (E), CYP2C19 (F), CYP2D6 (G), CYP2E1 (H), CYP3A4 ((I), MDZ), and CYP3A4 ((J), TST) in pooled HLM. The activity is expressed as the percentage of control samples containing no inhibitor (100%). Data show the mean ± standard deviation (SD) of three separate samples.
Figure 2
Figure 2
Changes in the inhibition curves of CYP2D6 by BBR (A) and THB (B) after pre-incubation of pooled HLM with (empty circle, ○) or without (solid circle, ●) NADPH for 30 min. The activity is expressed as the percentage of control samples containing no inhibitor (100%). Data show the mean ± SD of three separate samples.
Figure 3
Figure 3
Changes in the inhibition curves of CYP2D6 by TFD (A) and DMB (B) after pre-incubation of pooled HLM with (empty circle, ○) or without (solid circle, ●) NADPH for 30 min. The activity is expressed as the percentage of control samples containing no inhibitor (100%). Data show the mean ± SD of three separate samples.
Figure 4
Figure 4
Time-dependent inhibition of CYP2D6 by BBR. (A) BBR was incubated for 0, 1, 5, 10, 15, or 30 min with NADPH in pooled HLM. CYP2D6 activity is expressed as the log percentage of control group containing no inhibitor at zero time. Data show the mean ± SD for three separate samples. (B) Double-reciprocal plot of kobs and BBR concentrations was plotted to determine KI and kinact for the inactivation of CYP2D6 by BBR.
Figure 5
Figure 5
Reversibility of CYP inhibition by BBR (A), ketoconazole ((B)—a reversible inhibitor for CYP3A4), diltiazem ((C)—a quasi-irreversible inhibitor for CYP3A4), and mifepristone ((D)—an irreversible inhibitor for CYP3A4). BBR (5 μM), diltiazem (100 μM), or mifepristone (30 μM) was incubated for 30 min with NADPH in HLM, and microsomal protein was re-isolated by ultracentrifugation with (Ox + UF) or without (UF) CYP oxidation. Ketoconazole (1 μM) was incubated for 30 min without NADPH in HLM, and microsomal protein was re-isolated by ultracentrifugation (UF). The activity of re-isolated microsomal protein was compared with that with no manipulation (NM). Activity is expressed as the percentage of control samples containing no inhibitor (100%). Data show the mean ± SD for three separate samples.
Figure 6
Figure 6
Effects of BBR on nebivolol metabolism in pooled HLM. Nebivolol (1 μM) was incubated with pooled HLM (1 mg/mL) in the presence or absence of BBR at various concentrations for 60 min. Data are presented as the mean ± SD of three separate samples.
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References

    1. Elmer G.W., Lafferty W.E., Tyree P.T., Lind B.K. Potential interactions between complementary/alternative products and conventional medicines in a medicare population. Ann. Pharmacother. 2007;41:1617–1624. doi: 10.1345/aph.1K221. - DOI - PMC - PubMed
    1. Bjornsson T.D., Callaghan J.T., Einolf H.J., Fischer V., Gan L., Grimm S., Kao J., King S.P., Miwa G., Ni L. The conduct of In Vitro and In Vivo drug-drug interaction studies: A pharmaceutical research and manufacturers of America (phrma) perspective. Drug Metab. Dispos. 2003;31:815–832. doi: 10.1124/dmd.31.7.815. - DOI - PubMed
    1. Dresser G.K., Spence J.D., Bailey D.G. Pharmacokinetic-pharmacodynamic consequences and clinical relevance of cytochrome p450 3a4 inhibition. Clin. Pharmacokinet. 2000;38:41–57. doi: 10.2165/00003088-200038010-00003. - DOI - PubMed
    1. Levy R., Hachad H., Yao C., Ragueneau-Majlessi I. Relationship between extent of inhibition and inhibitor dose: Literature evaluation based on the metabolism and transport drug interaction database. Curr. Drug Metab. 2003;4:371–380. doi: 10.2174/1389200033489325. - DOI - PubMed
    1. Shaikh A.S., Thomas A.B., Chitlange S.S. Herb-drug interaction studies of herbs used in treatment of cardiovascular disorders—A narrative review of preclinical and clinical studies. Phytother. Res. 2020;34:1008–1026. doi: 10.1002/ptr.6585. - DOI - PubMed

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