title: Identification and characterization of potent CYP3A4 inhibitors in Schisandra fruit extract.
abstract: Schisandra fruit, a Schisandraceae family herb, is used as a component in Kampo medicines (developed from Chinese medicines, but established in Japan). It can act as a sedative and antitussive, improve hepatic function, and give a general tonic effect. An extract of Schisandra fruit has been shown with a potent inhibitory effect on human liver microsomal erythromycin N-demethylation activity mediated by cytochrome P450 3A4 (CYP3A4). The present study was conducted to identify Schisandra fruit components having inhibitory effects on CYP3A4 by surveying the effect on human liver microsomal erythromycin N-demethylation activity. Known components of Schisandra fruit, gomisins B, C, G, and N and gamma-shizandrin, showed inhibitory effects on N-demethylation activity. Among these components, gomisin C displayed the most potent and competitive inhibitory effect, with a Ki value of 0.049 microM. Furthermore, the inhibitory effect of gomisin C was stronger than that of ketoconazole (Ki = 0.070 microM), a known potent CYP3A4 inhibitor. Gomisin C, however, inhibited CYP1A2-, CYP2C9-, CYP2C19-, and CYP2D6-dependent activities only to a limited extent (IC50 values >10 microM). Moreover, gomisin C inactivated human liver microsomal erythromycin N-demethylation activity in a time- and concentration-dependent manner. The inactivation kinetic parameters k(inact) and K(I) were 0.092 min(-1) and 0.399 microM, respectively. The human liver microsomal erythromycin N-demethylation activity inactivated by gomisin C did not recover on dialysis of the microsomes. Spectral scanning of CYP3A4 with gomisin C yielded an absorbance at 455 nm, suggesting that gomisin C inactivated the cytochrome P450 via the formation of a metabolite intermediate complex. This pattern is consistent with the metabolism of the methylenedioxy substituent in gomisin C. These results indicate that gomisin C is a mechanism-based inhibitor that not only competitively inhibits but irreversibly inactivates CYP3A4.
authors.affiliation: Division of Natural Products Chemistry, Institute of Natural Medicine, Toyama Medical and Pharmaceutical University, 2630 Sugitani, Toyama, 930-0194, Japan.
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Iwata
authors.forename: Yasuhiro
authors.initials: Y
authors.lastname: Tezuka
authors.forename: Shigetoshi
authors.initials: S
authors.lastname: Kadota
authors.forename: Akira
authors.initials: A
authors.lastname: Hiratsuka
authors.forename: Tadashi
authors.initials: T
authors.lastname: Watabe
mesh_terms: D000900:Anti-Bacterial Agents / Q000378:metabolism; D002851:Chromatography, High Pressure Liquid; D034242:Cyclooctanes / Q000494:pharmacology; D051544:Cytochrome P-450 CYP3A; D065607:Cytochrome P-450 Enzyme Inhibitors; D004149:Dioxoles / Q000494:pharmacology; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D004917:Erythromycin / Q000378:metabolism; D005260:Female; D006801:Humans; D066298:In Vitro Techniques; D007654:Ketoconazole / Q000494:pharmacology; D017705:Lignans / Q000494:pharmacology; D009682:Magnetic Resonance Spectroscopy; D008297:Male; D013058:Mass Spectrometry; D017551:Microdialysis; D008862:Microsomes, Liver / Q000187:drug effects / Q000378:metabolism; D009904:Optical Rotation; D010936:Plant Extracts / Q000494:pharmacology; D011083:Polycyclic Compounds / Q000494:pharmacology; D032141:Schisandra / Q000737:chemistry*; D012997:Solvents
chemical_list: D000900:Anti-Bacterial Agents; D034242:Cyclooctanes; D065607:Cytochrome P-450 Enzyme Inhibitors; D004149:Dioxoles; D004791:Enzyme Inhibitors; D017705:Lignans; D010936:Plant Extracts; D011083:Polycyclic Compounds; D012997:Solvents; C082090:schisantherin B; D004917:Erythromycin; C034557:schizandrer A; C104464:CYP3A protein, human; D051544:Cytochrome P-450 CYP3A; C510163:CYP3A4 protein, human; D007654:Ketoconazole