Sridhar Vemulapalli


Cytochrome P450 3A4 (CYP3A4) and P-glycoprotein (P-gp) play an important role in the first pass metabolism thereby limits the oral bioavailability of many clinically important and frequently prescribed drugs. The absolute oral bioavailability of saquinavir is very low (i. e. 4%) due to its extensive first pass metabolism by the major metabolizing isozyme CYP3A4 and it is also a substrate of P-gp. Pomegranate juice (PGJ) was known to be a modulator of CYP3A4 and P-gp. Therefore, the aim of this study was to evaluate the influence of PGJ on the pharmacokinetics (PK) of saquinavir in wistar rats and on the P-gp mediated intestinal transport of saquinavir in everted gut sacs ex vivo.  Rats were treated orally with saquinavir (100 mg/kg) alone and in combination with PGJ (0.5, 1.0 and 2.0 mL/200g, body weight) for 15 consecutive days. Blood samples were collected on 1st day in single dose pharmacokinetic study (SDS) and on 15th day in multiple dose pharmacokinetic study (MDS). The peak plasma concentration (Cmax)and area under the plasma concentration-time curve (AUC0-24) of saquinavir was increased with PGJ in SDS (p<0.001) may be due to inhibition of CYP3A4 and P-gp. But interestingly, the Cmax and AUC0-24 of saquinavir was decreased significantly with PGJ in MDS. This is may be due to induction of CYP3A4. The transport of saquinavir was increased in presence of PGJ and known P-gp inhibitors (Verapamil, Ketoconazole and Quinindine) across the rat everted gut sacs ex vivo. The present study results suggested that PGJ has both effects (inhibition, in SDS and induction, in MDS) on CYP3A4-mediated saquinavir metabolism in vivo and inhibitory effect on the P-gp mediated intestinal transport of saquinavir ex vivo. Further studies are needed to confirm this interaction at cellular level using cell lines and in humans.


CYP3A4; P-glycoprotein; Everted gut sacs; Pomegranate juice; Pharmacokinetics

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Adukondalu D., Shravan KY., Vamshi VY., Shiva KR., Madhusudan RY., 2010. Effect of pomegranate juice pre-treatment on the transport of carbamazepine across rat intestine. Daru.18: 254-259.

Basu A., Penugonda K., 2009. Pomegranate juice: a heart-healthy fruit juice. Nutr Rev. 67: 49-56.

Bell C, Hawthorne S., 2008. Ellagic acid, pomegranate and prostate cancer—a mini review. J Pharm Pharmacol. 60: 139-144.

Beverly K., Matthew T., Dhiren RT., 2006. Deconvoluting the effects of P-glycoprotein on intestinal CYP3A: a major challenge. Current Opinion in Pharmacology. 6: 528-532.

Brian KA., Pamala AJ., Robin LC., Wayne AK., Michael EE., Bradley RW., Joseph GB., 2013. Koda-Kimble and Young’s applied therapeutics: The clinical use of drugs. Philadelphia: Lippincott Williams & Wilkins-Wolters Kluwer, United States of America.

Chandra P., Brouwer KL., 2004. The complexities of hepatic drug transport: current knowledge and emerging concepts. Pharm Res. 21: 719-735.

Dan LL., Anthony SF., Dennis LK., Stephen LH., Jameson JL., Joseph L., 2012. Harrison’s Principles of Internal Medicine. New York: McGraw-Hill Medical, United States of America.

Doherty MM., Charman WN., 2002. The mucosa of the small intestine: how clinically relevant as an organ of drug metabolism? Clin Pharmacokinet. 41: 235-253.

Doherty MM., Pang KS., 1997. First-pass effect: significance of the intestine for absorption and metabolism. Drug Chem Toxicol. 20: 329-344.

Eagling VA., Back DJ., Barry MG., 1997. Differential inhibition of cytochrome P450 isoforms by the protease inhibitors, ritonavir, saquinavir, and indinavir. Br J Clin Pharmacol. 44: 190-194.

Eberl S., Renner B., Neubert A., 2007. Role of p-glycoprotein inhibition for drug interactions: evidence from in vitro and pharmacoepidemiological studies. Clin Pharmacokinet. 46: 1039-1049.

Faria A., Monteiro R., Azevedo I., Calhau C., 2007. Pomegranate juice effects on cytochrome P450S expression: in vivo studies. J Med Food. 10: 643–649.

Farkas D., Oleson LE., Zhao Y., 2007. Pomegranate juice does not impair clearance of oral or intravenous midazolam, a probe for cytochrome P450-3A activity: comparison with grapefruit juice. J Clin Pharmacol. 47: 286–294.

Frappier S., Breilh D., Diarte E., Ba B., Ducint D., Pellegrin JL., Saux MC., 1998. Simultaneous determination of ritonavir and saquinavir, two human immunodeficiency virus protease inhibitors, in human serum by high-performance liquid chromatography. Journal of Chromatography B. 714: 384–389.

Genser D., 2008. Food and drug interaction: consequence for the nutrition/health status. Ann Nutr Metab. 52: S29–S32.

Hall SD., Thummel KE., Watkins PB., Lown KS., Benet LZ., Paine MF., Mayo RR., Turgeon DK., Bailey DG., Fontana RJ., 1999. Molecular and physical mechanisms of first-pass extraction. Drug Metab Dispos. 27: 161-166.

Harumi Y., Hajime K., Tetsuya N., Aikichi I., 2001. Simultaneous determination of the HIV protease inhibitors indinavir, amprenavir, saquinavir, ritonavir and nelfinavir in human plasma by high-performance liquid chromatography. Journal of Chromatography B. 755: 85–89.

Hidaka M., Okumura M., Fujita K., 2005. Effects of pomegranate juice on human cytochrome p450 3A (CYP3A) and carbamazepine pharmacokinetics in rats. Drug Metab Dispos. 33: 644–648.

Ho RH., Kim RB., 2005. Transporters and drug therapy: implications for drug disposition and disease. Clin Pharmacol Ther. 78: 260-277.

Holladay JW., Dewey MJ., Michniak BB., Wiltshire H., Halberg DL., Weigl P., Liang Z., Halifax K., Lindup WE., Back DJ., 2001. Elevated α1-acid glycoprotein reduces the volume of distribution and systemic clearance of saquinavir. Drug Metab Dispos. 29: 299-303.

Huang SM., Strong JM., Zhang L., Reynolds KS., Nallani S., Temple R., 2008. New era in drug interaction evaluation: US Food and Drug Administration update on CYP enzymes, transporters, and the guidance process. J Clin Pharmacol. 48: 662–670.

Huy RH., Ferenc F., Yvonne B., Stephan K., 1997. Determination of saquinavir in human plasma by high-performance liquid chromatography. Journal of Chromatography B. 694: 427–433.

Joseph TD., Robert LT., Gary CY., Gary RM., Barbara GW., Michael PL., 2008. Pharmacotherapy A Pathophysiologic Approach. New York: McGraw-Hill Medical, United States of America.

Jurenka JS., 2008. Therapeutic applications of pomegranate (Punica granatum L.): a review. Altern Med Rev. 13: 128–144.

Kenneth ET., Kent LK., Danny DS., 1997. Enzyme-catalyzed processes of first-pass hepatic and intestinal drug extraction. Advanced Drug Delivery Reviews. 27: 99-127.

Krueger DA., 2012. Composition of pomegranate juice. J AOAC Int. 95 (1): 163-168.

Kupferschmidt HH., Fattinger KE., Ha HR., Follath F., Krahenbuhl S., 1998. Grapefruit juice enhances the bioavailability of the HIV protease inhibitor saquinavir in man. Br J Clin Pharmacol. 45: 355–359.

Lansky EP., Newman RA., 2007. Punica granatum (pomegranate) and its potential for prevention and treatment of inflammation and cancer. J Ethnopharmacol. 109: 177–206.

Lee CG., Gottesman MM., Cardarelli CO., 1998. HIV-1 protease inhibitors are substrates for the MDR1 multidrug transporter. Biochemistry. 37: 3594–3601.

Letendre L., Scott M., Dobson G., Hidalgo I., Aungst B., 2004. Evaluating barriers to bioavailability in vivo: validation of a technique for separately assessing gastrointestinal absorption and hepatic extraction. Pharmaceutical Research. 21: 1457-1462.

Martin FM., Burhenne J., Ding R., 2002. Dose-dependent increase of saquinavir bioavailability by the pharmaceutic aid cremophor EL. British Journal of Clinical Pharmacology. 53: 576-581.

Michael DS., Melanie EE., Gerdi W., Jennifer JD., Mailine HC., Ruth M., Caren JR., 2005. Effects of Pomegranate Juice Consumption on Myocardial Perfusion in Patients With Coronary Heart Disease. The American Journal of Cardiology. 96; 810-814.

Muneaki H., Manabu O., Kenichi F., Tetsuya O., Keishi Y., Tomomi I., Nao S., Kazuhiko A., 2005. Effects of pomegranate juice on human cytochrome p450 3a (cyp3a) and carbamazepine pharmacokinetics in rats. Drug metabolism and disposition. 33: 644–648.

Nagata M,, Hidaka M,, Sekiya H., 2002. Effects of pomegranate juice on human cytochrome P450 2C9 and tolbutamide pharmacokinetics in rats. Drug Metab Dispos. 35: 302–305.

Neirinckx E., Vervaet C., De Boever S., Remon JP., Gommeren K., Daminet S., De Backer P., Croubels S., 2010. Species comparison of oral bioavailability, first-pass metabolism and pharmacokinetics of acetaminophen. Research in Veterinary Science. 89: 113-119.

Neurath AR., Strick N., Li YY., Debnath AK., 2005. Punica granatum(pomegranate) juice provides an HIV-1 entry inhibitor and candidate topical microbicide. Ann N Y Acad Sci. 1056: 311–327.

Noble S., Faulds D., 1996. Saquinavir. A review of its pharmacology and clinical potential in the management of HIV infection. Drugs. 52: 93–112.

Paine MF., Hart HL., Ludington SS., Haining RL., Rettie AE., Zeldin DC., 2006. The human intestinal cytochrome P450 ‘‘pie’’. Drug Metab Dispos. 34: 880-886.

Paine MF., Khalighi M., Fisher JM., Shen DD., Kunze KL., Marsh CL., Perkins JD., Thummel KE., 1997. Characterization of interintestinal and intraintestinal variations in human CYP3A-dependent metabolism. J Pharmacol Exp Ther. 283: 1552-1562.

Ravindra BP., Naveen BK., Haroled PL., Rajesh K., Jawahar BP., 2013. Influence of quercetin on the pharmacokinetics of ranolazine in rats and in vitro models. Drug Development and Industrial Pharmacy. 39: 873-879.

Shih HF., Yu CH., Pei DLC., 2005. Pharmacokinetic and pharmacodynamic interactions of morin and cyclosporine. Toxicology and Applied Pharmacology. 205: 65–70.

Shravan KY., Adukondalu D., Bhargavi LA., Vamshi VY., Ramesh G., Shiva KR., Madhusudan RY., Sarangapani M., 2011. Effect of pomegranate pretreatment on the oral bioavailability of buspirone in male albino rabbits. Daru. 19: 266-269.

Shriram MP., Ranjith KA., Subramanian G., Udupa N., 2007. Development and validation of a reversed-phase liquid chromatographic method with fluorescence detection for the study of Saquinavir pharmacokinetics in rat plasma. Analytica Chimica Acta. 594: 248–256.

Sridhar V., Surya SM., Ravindra BP., Naveen BK., 2014. Evaluation of First-Pass Cytochrome P4503A (CYP3A) and P-glycoprotein Activities Using Felodipine and Hesperetin in Combination in Wistar Rats and Everted Rat Gut Sacs in Vitro. Phytother. Res. 28: 699-705.

Stephane JM., Mary FP., Paul BW., 2004. Contributions of CYP3A4, P-glycoprotein, and Serum Protein Binding to the Intestinal First-Pass Extraction of Saquinavir. The journal of pharmacology and experimental therapeutics. 308: 941–948.

Swathi V., Shravan KY., Shiva KR., Ramesh G., Madhusudan RY., 2012. Effect of Pomegranate Juice on Intestinal Transport and Pharmacokinetics of Nitrendipine in Rats. Phytother. Res. 26: 1240–1245.

Undevia SD., Gomez AG., Ratain MJ., 2005. Pharmacokinetic variability of anticancer agents. Nat Rev Cancer. 5: 447–458.

Wacher VJ., Silverman JA., Zhang Y., Benet LZ., 1998. Role of P-glycoprotein and cytochrome P450 3A in limiting oral absorption of peptides and peptidomimetics. J Pharm Sci. 87: 1322-1330.

Wahajuddin., Raju KS., Singh SP., Taneja I., 2014. Investigation of the functional role of P-glycoprotein in limiting the oral bioavailability of lumefantrine. Antimicrob Agents Chemother. 58: 489-494.

Williams PEO., Muirhead GJ., Madigan MJ., Mitchell AM., Shaw T., 1992. Disposition and bioavailability of the HIV-proteinase inhibitor, Ro 31–8959, after single doses in healthy volunteers. Br J Clin Pharmacol. 34:155P–156P.

Yang J., Jamei M., Yeo KR., Tucker GT., Rostami HA., 2007. Prediction of intestinal first-pass drug metabolism. Curr Drug Metab. 8: 676–84.



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