Mechanism-Based Inactivation of Human Liver Microsomal CYP3A4 by Rutaecarpine and Limonin from Evodia Fruit Extract

Abstract

Evodia fruit (Evodiae Fructus) is used as a herbal medicine prepared from the matured fruit of the Evodia rutaecarpa Bentham or Evodia officinalis Dode, of the Rutaceae plant family. An extract of Evodia fruit in the presence of NADPH was shown to inhibit human liver microsomal erythromycin N-demethylation activity, mediated by cytochrome P450 3A4 (CYP3A4), in a preincubation-time dependent manner. The present study was conducted to identify components of Evodia fruit extract having preincubation-time dependent inhibitory effects on CYP3A4 by analyzing human liver microsomal erythromycin N-demethylation activity. Rutaecarpine, a major component of Evodia fruit, and limonin caused the most dramatic decrease in residual CYP3A4 activity (IC50 before and after 20 min preincubation with: rutaecarpine, >100 microM and 1.4 microM; limonin, 23.5 microM and 1.8 microM, respectively). Furthermore, rutaecarpine and limonin were identified as mechanism-based inhibitors of CYP3A4 from the following observations: 1) The inhibitory effects of rutaecarpine and limonin on CYP3A4 activity were dependent on the preincubation time, 2) The inhibition required NADPH, 3) The inhibition was depressed in the presence of the competitive CYP3A4 inhibitor, ketoconazole, 4) Dialysis resulted in no recovery of CYP3A4 activity. The kinetic parameters for inactivation k(inact) and K(I) were: 0.387 min-1 and 107.7 microM for rutaecarpine, 0.266 min-1 and 23.2 microM for limonin, respectively. These results indicate that rutaecarpine and limonin are mechanism-based inhibitors of CYP3A4.

Rutaecarpine Induces Chloride Secretion across Rat Isolated Distal Colon

“…In addition, Fructus evodiae (fruit of E. rutaecarpa) is also used for the treatment of headache, thoracicoabdominal pain, vomiting, colds, and reduced blood circulation (Fei et al, 2003). Pharmacological studies indicate that Rut has various bioactivities, such as causing vasodilatation by mechanisms of an endothelium-dependent manner coupled with the synthesis or release of nitric oxide (Wang et al, 1996), inhibiting vasoconstriction induced by anaphylaxis (Yu et al, 2005), inducing positive inotropic and chronotropic actions (Kobayashi et al, 2001), protecting the myocardium mediated by calcitonin gene-related peptide (Hu et al, 2002;Yi et al, 2004), increasing intracellular Ca 2ϩ concentration in endothelium (Wang et al, 1996), suppressing platelet plug formation in mesenteric venules (Sheu et al, 2000), inhibiting prostaglandin production in RAW264.7 macrophages (Woo et al, 2001), relaxation of rabbit and human internal anal sphincter (Jiang et al, 2000), gastroprotective effect against injury induced by aspirin and stress (Wang et al, 2005), anti-Helicobacter pylori (Tominaga et al, 2002), blockade of delayed rectifier K ϩ current in NG108-15 neuronal cells (Wu et al, 2001), inhibiting cytochrome P450 in human liver microsomes (Ueng et al, 2002(Ueng et al, , 2006Iwata et al, 2005), and inhibition of COX-2 (Moon et al, 1999).…”

Identification and Characterization of Potent CYP2B6 Inhibitors in Woohwangcheongsimwon Suspension, an Herbal Preparation Used in the Treatment and Prevention of Apoplexy in Korea and China

“…In human liver microsomes with a NADPH-generating system, Woohwangcheongsimwon suspension moderately inhibited metabolic CYP2B6-catalyzed bupropion hydroxylase activity, regardless of preincubation with microsomes, indicating that a mechanism-based inhibitory component was not present in Woohwangcheongsimwon suspension (Fig. 1) (Iwata et al, 2005). Woohwangcheongsimwon also inhibited CYP2B6 activity with IC 50 values of 110 and 190 g/ml for bupropion hydroxylase and efavirenz 8-hydroxylase activity, respectively.…”

Mechanism-Based Inhibition of Recombinant Human Cytochrome P450 3A4 by Tomato Juice Extract

“…benzopyran-7-one) furanocumarins from grapefruit, 18) rutaecarpine and limonene from Evodia rutaecarpa, 19) methylenedioxyphenyl lignans from Piper, 20) kaempferol from Zingiber aromaticum, 21) 5-methoxypsoralen from Foeniculum vulgare, 22) and lignans from Phyllanthus amarus, 23) have all been shown to be responsible for MBI of CYP3A4. Moreover, interactions between these compounds and therapeutic drugs could occur in vivo.…”

“…[14][15][16] Mechanism-based inhibition (MBI) of CYP3A4 is characterized by nicotinamide adenine dinucleotide phosphate (NADPH)-, time-, and concentration-dependent enzyme inactivation that occurs when some substrates are converted by CYPs into reactive metabolites. 17) Several phytochemicals, including GF-I-1 (4-[ [6-hydroxy-7 [1] benzopyran-7-one) furanocumarins from grapefruit, 18) rutaecarpine and limonene from Evodia rutaecarpa, 19) methylenedioxyphenyl lignans from Piper, 20) kaempferol from Zingiber aromaticum, 21) 5-methoxypsoralen from Foeniculum vulgare, 22) and lignans from Phyllanthus amarus, 23) have all been shown to be responsible for MBI of CYP3A4. Moreover, interactions between these compounds and therapeutic drugs could occur in vivo.…”