In Vitro and In Vivo Evaluation of the Effect of Puerarin on Hepatic Cytochrome P450-Mediated Drug Metabolism

Abstract

Puerarin (8-β-D-glucopyranosyl-7-hydroxy-3-(4-hydroxyphenyl)-4H-1-benzopyran-4-one) is a major pharmacological component of Puerariae Radix, the root of Pueraria lobata. We investigated the effect of puerarin on hepatic cytochrome P450-mediated drug metabolism in rats and humans. The in vitro cytochrome P450 inhibitory effect of puerarin in human and rat liver microsomes was evaluated using the following model cytochrome P450 substrates: phenacetin for CYP1A, diclofenac for CYP2C, dextromethorphan for CYP2D, and testosterone for CYP3A. The in vivo pharmacokinetics of intravenous and oral buspirone, a probe substrate for CYP3A, was studied with single simultaneous intravenous coadministration of puerarin in rats. In the in vitro cytochrome P450 inhibition study, the rate of disappearance of testosterone was significantly reduced in the presence of 10 µM PU, while that of other cytochrome P450 substrates was not significantly affected in both human and rat liver microsomes, suggesting that puerarin inhibits the in vitro hepatic CYP3A-mediated metabolism in the human and rat systems (IC₅₀ = 15.5 ± 3.9 µM). After intravenous administration of buspirone with single simultaneous coadministration of intravenous puerarin at a dose of 10 mg/kg in rats, the total area under the plasma concentration–time curve from time zero to time infinity was increased while time-averaged total body clearance decreased. When buspirone was orally administered in rats with the 10 mg/kg intravenous puerarin coadministration, both total area under the plasma concentration–time curve from time zero to time infinity and the extent of absolute oral bioavailability were significantly increased. Therefore, results of the in vitro microsomal and in vivo pharmacokinetic studies suggest the possible inhibition of hepatic CYP3A-mediated drug metabolism by puerarin administration, potentially leading to metabolism-mediated herb–drug interactions with clinical significance.

Key words

puerarin - Pueraria lobata - Fabaceae - cytochrome P450 - buspirone - hepatic metabolism - pharmacokinetics

^* Sang-Bum Kim and In-Soo Yoon contributed equally to this work.

• References

• 1 Bent S. Herbal medicine in the United States: review of efficacy, safety, and regulation: grand rounds at University of California, San Francisco Medical Center. J Gen Intern Med 2008; 23: 854-859
• 2 Bardia A, Nisly NL, Zimmerman MB, Gryzlak BM, Wallace RB. Use of herbs among adults based on evidence-based indications: findings from the National Health Interview Survey. Mayo Clin Proc 2007; 82: 561-566
• 3 Kennedy DA, Seely D. Clinically based evidence of drug-herb interactions: a systematic review. Expert Opin Drug Saf 2010; 9: 79-124
• 4 Zhou S, Gao Y, Jiang W, Huang M, Xu A, Paxton JW. Interactions of herbs with cytochrome P450. Drug Metab Rev 2003; 35: 35-98
• 5 Rendic S. Summary of information on human CYP enzymes: human P450 metabolism data. Drug Metab Rev 2002; 34: 83-448
• 6 Girennavar B, Jayaprakasha GK, Patil BS. Potent inhibition of human cytochrome P450 3A4, 2D6, and 2C9 isoenzymes by grapefruit juice and its furocoumarins. J Food Sci 2007; 72: C417-C421
• 7 Tassaneeyakul W, Guo LQ, Fukuda K, Ohta T, Yamazoe Y. Inhibition selectivity of grapefruit juice components on human cytochromes P450. Arch Biochem Biophys 2000; 378: 356-363
• 8 Mason P. Important drug-nutrient interactions. Proc Nutr Soc 2010; 69: 551-557
• 9 Komoroski BJ, Zhang S, Cai H, Hutzler JM, Frye R, Tracy TS, Strom SC, Lehmann T, Ang CY, Cui YY, Venkataramanan R. Induction and inhibition of cytochromes P450 by the St. Johnʼs wort constituent hyperforin in human hepatocyte cultures. Drug Metab Dispos 2004; 32: 512-518
• 10 Mohamed MF, Frye RF. Inhibition of intestinal and hepatic glucuronidation of mycophenolic acid by Ginkgo biloba extract and flavonoids. Drug Metab Dispos 2010; 38: 270-275
• 11 Yin OQ, Tomlinson B, Waye MM, Chow AH, Chow MS. Pharmacogenetics and herb-drug interactions: experience with Ginkgo biloba and omeprazole. Pharmacogenetics 2004; 14: 841-850
• 12 Wang Q, Wu T, Chen X, Ni J, Duan X, Zheng J, Qiao J, Zhou L, Wei J. Puerarin injection for unstable angina pectoris. Cochrane Database Syst Rev 2006; (3) CD004196
• 13 Luo CF, Cai B, Hou N, Yuan M, Liu SM, Ji H, Xiong LG, Xiong W, Luo JD, Chen MS. UDP-glucuronosyltransferase 1A1 is the principal enzyme responsible for puerarin metabolism in human liver microsomes. Arch Toxicol 2012; 86: 1681-1690
• 14 Zheng J, Chen B, Jiang B, Zeng L, Tang ZR, Fan L, Zhou HH. The effects of puerarin on CYP2D6 and CYP1A2 activities in vivo . Arch Pharm Res 2010; 33: 243-246
• 15 Guerra MC, Speroni E, Broccoli M, Cangini M, Pasini P, Minghett A, Crespi-Perellino N, Mirasoli M, Cantelli-Forti G, Paolini M. Comparison between chinese medical herb Pueraria lobata crude extract and its main isoflavone puerarin antioxidant properties and effects on rat liver CYP-catalysed drug metabolism. Life Sci 2000; 67: 2997-3006
• 16 Foti RS, Rock DA, Wienkers LC, Wahlstrom JL. Selection of alternative CYP3A4 probe substrates for clinical drug interaction studies using in vitro data and in vivo simulation. Drug Metab Dispos 2010; 38: 981-987
• 17 Ohno Y, Hisaka A, Suzuki H. General framework for the quantitative prediction of CYP3A4-mediated oral drug interactions based on the AUC increase by coadministration of standard drugs. Clin Pharmacokinet 2007; 46: 681-696
• 18 Rioux N, Bellavance E, Bourg S, Garneau M, Ribadeneira MD, Duan J. Assessment of CYP3A-mediated drug-drug interaction potential for victim drugs using an in vivo rat model. Biopharm Drug Dispos 2013; 34: 396-401
• 19 Zhu L, Yang X, Zhou J, Tang L, Xia B, Hu M, Zhou F, Liu Z. The exposure of highly toxic aconitine does not significantly impact the activity and expression of cytochrome P450 3A in rats determined by a novel ultra performance liquid chromatography-tandem mass spectrometric method of a specific probe buspirone. Food Chem Toxicol 2013; 51: 396-403
• 20 Tassaneeyakul W, Birkett DJ, Veronese ME, McManus ME, Tukey RH, Quattrochi LC, Gelboin HV, Miners JO. Specificity of substrate and inhibitor probes for human cytochromes P450 1A1 and 1A2. J Pharmacol Exp Ther 1993; 265: 401-407
• 21 Leemann T, Transon C, Dayer P. Cytochrome P450 TB (CYP2C): a major monooxygenase catalyzing diclofenac 4′-hydroxylation in human liver. Life Sci 1993; 52: 29-34
• 22 Zhao K, Ding M, Cao H, Cao ZX. In-vitro metabolism of glycyrrhetinic acid by human and rat liver microsomes and its interactions with six CYP substrates. J Pharm Pharmacol 2012; 64: 1445-1451
• 23 Transon C, Leemann T, Dayer P. In vitro comparative inhibition profiles of major human drug metabolising cytochrome P450 isozymes (CYP2C9, CYP2D6 and CYP3A4) by HMG-CoA reductase inhibitors. Eur J Clin Pharmacol 1996; 50: 209-215
• 24 Eagling VA, Tjia JF, Back DJ. Differential selectivity of cytochrome P450 inhibitors against probe substrates in human and rat liver microsomes. Br J Clin Pharmacol 1998; 45: 107-114
• 25 Knutter I, Kottra G, Fischer W, Daniel H, Brandsch M. High-affinity interaction of sartans with H+/peptide transporters. Drug Metab Dispos 2009; 37: 143-149
• 26 Hari Kumar KB, Kuttan R. Inhibition of drug metabolizing enzymes (cytochrome P450) in vitro as well as in vivo by Phyllanthus amarus SCHUM & THONN. Biol Pharm Bull 2006; 29: 1310-1313
• 27 Gammans RE, Mayol RF, LaBudde JA. Metabolism and disposition of buspirone. Am J Med 1986; 80: 41-51
• 28 Mahmood I, Sahajwalla C. Clinical pharmacokinetics and pharmacodynamics of buspirone, an anxiolytic drug. Clin Pharmacokinet 1999; 36: 277-287
• 29 Yoon I, Han S, Choi YH, Kang HE, Cho HJ, Kim JS, Shim CK, Chung SJ, Chong S, Kim DD. Saturable sinusoidal uptake is rate-determining process in hepatic elimination of docetaxel in rats. Xenobiotica 2012; 42: 1110-1119
• 30 Obach RS, Walsky RL, Venkatakrishnan K, Gaman EA, Houston JB, Tremaine LM. The utility of in vitro cytochrome P450 inhibition data in the prediction of drug-drug interactions. J Pharmacol Exp Ther 2006; 316: 336-348
• 31 Yan B, Wang W, Zhang L, Xing D, Wang D, Du L. Determination of puerarin in rat cortex by high-performance liquid chromatography after intravenous administration of Puerariae flavonoids. Biomed Chromatogr 2006; 20: 180-184
• 32 Jin XL, Zhu XY. Pharmacokinetics of puerarin in rats, rabbits, and dogs. Zhongguo Yao Li Xue Bao 1992; 13: 284-288
• 33 Kim JE, Cho HJ, Kim JS, Shim CK, Chung SJ, Oak MH, Yoon IS, Kim DD. The limited intestinal absorption via paracellular pathway is responsible for the low oral bioavailability of doxorubicin. Xenobiotica 2013; 43: 579-591
• 34 Jurica J, Konecny J, Zahradnikova LZ, Tomandl J. Simultaneous HPLC determination of tolbutamide, phenacetin and their metabolites as markers of cytochromes 1A2 and 2C6/11 in rat liver perfusate. J Pharm Biomed Anal 2010; 52: 557-564
• 35 Reyes-Gordillo K, Muriel P, Castaneda-Hernandez G, Favari L. Pharmacokinetics of diclofenac in rats intoxicated with CCL4, and in the regenerating liver. Biopharm Drug Dispos 2007; 28: 415-422
• 36 Zhang R, Liu C, Wang N, Mi S. Determination of cytochrome P450 3A4 activity with testosterone probe using high performance liquid chromatography. Se Pu 2008; 26: 80-83