In vitro Interactions with Repeated Grapefruit Juice Administration – to Peel or not to Peel?

Abstract

Interactions of acutely administered grapefruit juice (GFJ) with cytochrome P450 isoform 3A4 (CYP3A4) and P-glycoprotein (Pgp) function are well established. In this study, we investigated in vitro the effect of repeated administration of GFJ and its major constituents (the flavonoid naringin, its aglycone naringenin and the furanocoumarin bergamottin) on mRNA expression of MDR1 and CYP3A4 in LS180 cells. Since the bergamottin content is higher in the peel than in the fruit, we compared GFJ containing peel (GFJP+) with juice without any peel extract (GFJP-). GFJP- (1 %) showed no significant effect on MDR1 and CYP3A4 mRNA expression, whereas 1 % GFJP+ increased expression of MDR1 3.7-fold (P < 0.01) and CYP3A4 2.3-fold (P < 0.05). Of the tested constituents, only 10 μM bergamottin and 200 μM naringenin induced MDR1 mRNA levels 2.9- and 4.0-fold, respectively (P < 0.01 for both), and CYP3A4 mRNA levels 3.2- and 15.6-fold (P < 0.01 for both), respectively. Western blot analysis and rhodamine 123 uptake experiments partly confirmed these findings on the protein and the functional level. In summary, GFJ containing no peel extract may have a lower potential for interactions with CYP3A4 or P-glycoprotein.

Key words

grapefruit juice - bergamottin - CYP3A4 - P-glycoprotein - Citrus x paradisi Macfad - naringin/naringenin

References

• 1 Bailey D G, Malcolm J, Arnold O, Spence J D. Grapefruit juice-drug interactions.  Br J Clin Pharmacol. 1998;  46 101-10
• 2 Lown K S, Bailey D G, Fontana R J, Janardan S K, Adair C H, Fortlage L H. et al . Grapefruit juice increases felodipine oral availability in humans by decreasing intestinal CYP3A protein expression.  J Clin Invest. 1997;  99 2545-53
• 3 Takanaga H, Ohnishi A, Matsuo H, Sawada Y. Inhibition of vinblastine efflux mediated by P-glycoprotein by grapefruit juice components in caco-2 cells.  Biol Pharm Bull. 1998;  21 1062-6
• 4 Wang E J, Casciano C N, Clement R P, Johnson W W. Inhibition of P-glycoprotein transport function by grapefruit juice psoralen.  Pharm Res. 2001;  18 432-8
• 5 de Castro W V, Mertens-Talcott S, Derendorf H, Butterweck V. Grapefruit juice-drug interactions: Grapefruit juice and its components inhibit P-glycoprotein (ABCB1) mediated transport of talinolol in Caco-2 cells.  J Pharm Sci. 2007;  96 2808-17
• 6 Mertens-Talcott S U, De Castro W V, Manthey J A, Derendorf H, Butterweck V. Polymethoxylated flavones and other phenolic derivates from citrus in their inhibitory effects on P-glycoprotein-mediated transport of talinolol in Caco-2 cells.  J Agric Food Chem. 2007;  55 2563-8
• 7 Ducharme M P, Warbasse L H, Edwards D J. Disposition of intravenous and oral cyclosporine after administration with grapefruit juice.  Clin Pharmacol Ther. 1995;  57 485-91
• 8 Kupferschmidt H H, Ha H R, Ziegler W H, Meier P J, Krahenbuhl S. Interaction between grapefruit juice and midazolam in humans.  Clin Pharmacol Ther. 1995;  58 20-8
• 9 Schmiedlin-Ren P, Edwards D J, Fitzsimmons M E, He K, Lown K S, Woster P M. Mechanisms of enhanced oral availability of CYP3A4 substrates by grapefruit constituents. Decreased enterocyte CYP3A4 concentration and mechanism-based inactivation by furanocoumarins.  Drug Metab Dispos. 1997;  25 1228-33
• 10 De Castro W V, Mertens-Talcott S, Rubner A, Butterweck V, Derendorf H. Variation of flavonoids and furanocoumarins in grapefruit juices: a potential source of variability in grapefruit juice-drug interaction studies.  J Agric Food Chem. 2006;  54 249-55
• 11 Ho P C, Saville D J, Coville P F, Wanwimolruk S. Content of CYP3A4 inhibitors, naringin, naringenin and bergapten in grapefruit and grapefruit juice products.  Pharm Acta Helv. 2000;  74 379-85
• 12 Edwards K G, Stoker J R. Biosynthesis of coumarin: the isomerization stage.  Phytochemistry. 1967;  6 655-61
• 13 Paine M F, Criss A B, Watkins P B. Two major grapefruit juice components differ in intestinal CYP3A4 inhibition kinetic and binding properties.  Drug Metab Dispos. 2004;  32 1146-53
• 14 Fuhr U, Kummert A L. The fate of naringin in humans: a key to grapefruit juice-drug interactions.  Clin Pharmacol Ther. 1995;  58 365-73
• 15 Ubeaud G, Hagenbach J, Vandenschrieck S, Jung L, Koffel J C. In vitro inhibition of simvastatin metabolism in rat and human liver by naringenin.  Life Sci. 1999;  65 1403-12
• 16 Kane G C, Lipsky J J. Drug-grapefruit juice interactions.  Mayo Clin Proc. 2000;  75 933-42
• 17 Romiti N, Tramonti G, Donati A, Chieli E. Effects of grapefruit juice on the multidrug transporter P-glycoprotein in the human proximal tubular cell line HK-2.  Life Sci. 2004;  76 293-302
• 18 Skehan P, Storeng R, Scudiero D, Monks A, McMahon J, Vistica D. New colorimetric cytotoxicity assay for anticancer-drug screening.  J Natl Cancer Inst. 1990;  82 1107-12
• 19 Pfrunder A, Gutmann H, Beglinger C, Drewe J. Gene expression of CYP3A4, ABC-transporters (MDR1 and MRP1-MRP5) and hPXR in three different human colon carcinoma cell lines.  J Pharm Pharmacol. 2003;  55 59-66
• 20 Bogman K, Peyer A K, Török M, Küsters E, Drewe J. HMG-CoA reductase inhibitors and P-glycoprotein modulation.  Br J Pharmacol. 2001;  132 1183-92