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Effects of Commonly Used Excipients on the Expression of CYP3A4 in Colon and Liver Cells

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ABSTRACT

Purpose

The objective of this investigation was to assess whether common pharmaceutical excipients regulate the expression of drug-metabolizing enzymes in human colon and liver cells.

Methods

Nineteen commonly used excipients were evaluated using a panel of experiments including cell-based human PXR activation assays, real-time RT-PCR assays for CYP3A4 mRNA expression, and immunoblot analysis of CYP3A4 protein expression in immortalized human liver cells (HepG2 and Fa2N4), human primary hepatocytes, and the intestinal LS174T cell models.

Results

No excipient activated human PXR or practically induced CYP3A4. However, three excipients (polysorbate 80, pregelatinized starch, and hydroxypropyl methylcellulose) tended to decrease mRNA and protein expression across experimental models.

Conclusion

This study represents the first investigation of the potential role of excipients in the expression of drug-metabolizing enzymes. Findings imply that some excipients may hold potential for excipient-drug interactions by repression of CYP3A4 expression.

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Abbreviations

CA:

Citric acid

CCS:

croscarmellose sodium

CYP3A4:

cytochrome P450 3A4

DCP:

dicalcium phosphate dehydrate

DMSO:

dimethyl sulfoxide

FA:

fumaric acid

HPMC:

hydroxypropyl methylcellulose

KTC:

ketaconazole

LAC:

lactose

MA:

malic acid

MCC:

microcrystalline cellulose

MgStr:

magnesium stearate

MDR1:

multidrug resistance

PEG-3350:

polyethylene glycol 3350

PS-80:

polysorbate-80

PVP:

povidone

PXR:

pregnane X receptor

PgS:

pregelatinized starch

PG:

propylene glycol

RIF:

rifampicin

SD:

amorphous fumed silicon dioxide

SLS:

sodium lauryl sulfate

SSG:

sodium starch glycolate

SUC:

sucrose

X-PVP:

Crospovidone

REFERENCES

  1. FDA. Waiver of in vivo bioavailability and bioequivalence studies for immediate-release solid oral dosage forms based on a biopharmaceutics classification system. CDER, Guidance for Industry. 2000.

  2. EMEA. Note for guidance on the investigation of bioavailability and bioequivalence. Committee for Proprietary Medicinal Products. 2001.

  3. Polli JE, Yu LX, Cook JA, Amidon GL, Borchardt RT, Burnside BA, et al. Summary workshop report: biopharmaceutics classification system–implementation challenges and extension opportunities. J Pharm Sci. 2004;93:1375–81.

    Article  PubMed  CAS  Google Scholar 

  4. Polli JE, Abrahamsson BS, Yu LX, Amidon GL, Baldoni JM, Cook JA, et al. Summary workshop report: bioequivalence, biopharmaceutics classification system, and beyond. AAPS J. 2008;10:373–9.

    Article  PubMed  Google Scholar 

  5. Yu LX, Amidon GL, Polli JE, Zhao H, Mehta MU, Conner DP, et al. Biopharmaceutics classification system: the scientific basis for biowaiver extensions. Pharm Res. 2002;19:921–5.

    Article  PubMed  CAS  Google Scholar 

  6. Benet LZ, Amidon GL, Barends DM, Lennernas H, Polli JE, Shah VP, et al. The use of BDDCS in classifying the permeability of marketed drugs. Pharm Res. 2008;25:483–8.

    Article  PubMed  CAS  Google Scholar 

  7. EMEA. Guideline on the investigation of bioequivalence. Committee for Medicinal Products for Human Use. 2008.

  8. Koch KM, Parr AF, Tomlinson JJ, Sandefer EP, Digenis GA, Donn KH, et al. Effect of sodium acid pyrophosphate on ranitidine bioavailability and gastrointestinal transit time. Pharm Res. 1993;10:1027–30.

    Article  PubMed  CAS  Google Scholar 

  9. Adkin DA, Davis SS, Sparrow RA, Huckle PD, Wilding IR. The effect of mannitol on the oral bioavailability of cimetidine. J Pharm Sci. 1995;84:1405–9.

    Article  PubMed  CAS  Google Scholar 

  10. Wuand CY, Benet LZ. Predicting drug disposition via application of BCS: transport/absorption/ elimination interplay and development of a biopharmaceutics drug disposition classification system. Pharm Res. 2005;22:11–23.

    Article  CAS  Google Scholar 

  11. Klaassen CD, Slitt AL. Regulation of hepatic transporters by xenobiotic receptors. Curr Drug Metab. 2005;6:309–28.

    Article  PubMed  CAS  Google Scholar 

  12. Meyer UA. Overview of enzymes of drug metabolism. J Pharmacokinet Biopharm. 1996;24:449–59.

    Article  PubMed  CAS  Google Scholar 

  13. Lin JH. Sense and nonsense in the prediction of drug-drug interactions. Curr Drug Metab. 2000;1:305–31.

    Article  PubMed  CAS  Google Scholar 

  14. Wang H, LeCluyse EL. Role of orphan nuclear receptors in the regulation of drug-metabolising enzymes. Clin Pharmacokinet. 2003;42:1331–57.

    Article  PubMed  CAS  Google Scholar 

  15. Sachs-Barrable K, Thamboo A, Lee SD, Wasan KM. Lipid excipients Peceol and Gelucire 44/14 decrease P-glycoprotein mediated efflux of rhodamine 123 partially due to modifying P-glycoprotein protein expression within Caco-2 cells. J Pharm Pharm Sci. 2007;10:319–31.

    PubMed  CAS  Google Scholar 

  16. Barta CA, Sachs-Barrable K, Feng F, Wasan KM. Effects of monoglycerides on P-glycoprotein: modulation of the activity and expression in Caco-2 cell monolayers. Mol Pharm. 2008;5:863–75.

    Article  PubMed  CAS  Google Scholar 

  17. Kumar GN, Surapaneni S. Role of drug metabolism in drug discovery and development. Med Res Rev. 2001;21:397–411.

    Article  PubMed  CAS  Google Scholar 

  18. Wang H, Tompkins LM. CYP2B6: new insights into a historically overlooked cytochrome P450 isozyme. Curr Drug Metab. 2008;9:598–610.

    Article  PubMed  CAS  Google Scholar 

  19. Kliewer SA, Moore JT, Wade L, Staudinger JL, Watson MA, Jones SA, et al. An orphan nuclear receptor activated by pregnanes defines a novel steroid signaling pathway. Cell. 1998;92:73–82.

    Article  PubMed  CAS  Google Scholar 

  20. Lehmann JM, McKee DD, Watson MA, Willson TM, Moore JT, Kliewer SA. The human orphan nuclear receptor PXR is activated by compounds that regulate CYP3A4 gene expression and cause drug interactions. J Clin Investig. 1998;102:1016–23.

    Article  PubMed  CAS  Google Scholar 

  21. LeCluyse EL, Alexandre E, Hamilton GA, Viollon-Abadie C, Coon DJ, Jolley S, et al. Isolation and culture of primary human hepatocytes. Methods Mol Biol (Clifton, NJ). 2005;290:207–29.

    Google Scholar 

  22. Li L, Chen T, Stanton JD, Sueyoshi T, Negishi M, Wang H. The peripheral benzodiazepine receptor ligand 1-(2-chlorophenyl-methylpropyl)-3-isoquinoline-carboxamide is a novel antagonist of human constitutive androstane receptor. Mol Pharmacol. 2008;74:443–53.

    Article  PubMed  CAS  Google Scholar 

  23. Faucette SR, Sueyoshi T, Smith CM, Negishi M, Lecluyse EL, Wang H. Differential regulation of hepatic CYP2B6 and CYP3A4 genes by constitutive androstane receptor but not pregnane X receptor. J Pharmacol Exp Ther. 2006;317:1200–9.

    Article  PubMed  CAS  Google Scholar 

  24. Wang H, Faucette S, Sueyoshi T, Moore R, Ferguson S, Negishi M, et al. A novel distal enhancer module regulated by pregnane X receptor/constitutive androstane receptor is essential for the maximal induction of CYP2B6 gene expression. J Biol Chem. 2003;278:14146–52.

    Article  PubMed  CAS  Google Scholar 

  25. Li L, Stanton JD, Tolson AH, Luo Y, Wang H. Bioactive terpenoids and flavonoids from Ginkgo biloba extract induce the expression of hepatic drug-metabolizing enzymes through pregnane X receptor, constitutive androstane receptor, and aryl hydrocarbon receptor-mediated pathways. Pharm Res. 2009;26:872–82.

    Article  PubMed  CAS  Google Scholar 

  26. Habib YS, Augsburger LL, Shangraw RF. Production of inert cushioning beads: effect of excipients on the physicomechanical properties of freeze-dried beads containing microcrystalline cellulose produced by extrusion-spheronization. Int J Pharm. 2002;233:67–83.

    Article  PubMed  CAS  Google Scholar 

  27. Shangraw RF. Emerging trends in the use of pharmaceutical excipients. Pharm Technol. 1997;21:36–42.

    Google Scholar 

  28. Digenis GA, Gold TB, Shah VP. Cross-linking of gelatin capsules and its relevance to their in vitro-in vivo performance. J Pharm Sci. 1994;83:915–21.

    Article  PubMed  CAS  Google Scholar 

  29. Fetzner A, Bohm S, Schreder S, Schubert R. Degradation of raw or film-incorporated beta-cyclodextrin by enzymes and colonic bacteria. Eur J Pharm Biopharm. 2004;58:91–7.

    Article  PubMed  CAS  Google Scholar 

  30. Yamagata T, Kusuhara H, Morishita M, Takayama K, Benameur H, Sugiyama Y. Improvement of the oral drug absorption of topotecan through the inhibition of intestinal xenobiotic efflux transporter, breast cancer resistance protein, by excipients. Drug Metab Dispos. 2007;35:1142–8.

    Article  PubMed  CAS  Google Scholar 

  31. Yamagata T, Morishita M, Kusuhara H, Takayama K, Benameur H, Sugiyama Y. Characterization of the inhibition of breast cancer resistance protein-mediated efflux of mitoxantrone by pharmaceutical excipients. Int J Pharm. 2009;370:216–9.

    Article  PubMed  CAS  Google Scholar 

  32. Luo G, Cunningham M, Kim S, Burn T, Lin J, Sinz M, et al. CYP3A4 induction by drugs: correlation between a pregnane X receptor reporter gene assay and CYP3A4 expression in human hepatocytes. Drug Metab Dispos. 2002;30:795–804.

    Article  PubMed  CAS  Google Scholar 

  33. FDA. In vitro evaluation of CYP induction. FDA Guidelines. 2006.

  34. Burk O, Koch I, Raucy J, Hustert E, Eichelbaum M, Brockmoller J, et al. The induction of cytochrome P450 3A5 (CYP3A5) in the human liver and intestine is mediated by the xenobiotic sensors pregnane X receptor (PXR) and constitutively activated receptor (CAR). J Biol Chem. 2004;279:38379–85.

    Article  PubMed  CAS  Google Scholar 

  35. Glaeser H, Drescher S, Eichelbaum M, Fromm MF. Influence of rifampicin on the expression and function of human intestinal cytochrome P450 enzymes. Br J Clin Pharmacol. 2005;59:199–206.

    Article  PubMed  CAS  Google Scholar 

  36. Rane Y, Mashru R, Sankalia M, Sankalia J. Effect of hydrophilic swellable polymers on dissolution enhancement of carbamazepine solid dispersions studied using response surface methodology. AAPS PharmSciTech. 8:Article 27. 2007.

    Google Scholar 

  37. Rege BD, Kao JP, Polli JE. Effects of nonionic surfactants on membrane transporters in Caco-2 cell monolayers. Eur J Pharm Sci. 2002;16:237–46.

    Article  PubMed  CAS  Google Scholar 

  38. Rege BD, Yu LX, Hussain AS, Polli JE. Effect of common excipients on Caco-2 transport of low-permeability drugs. J Pharm Sci. 2001;90:1776–86.

    Article  PubMed  CAS  Google Scholar 

  39. Aitken AE, Morgan ET. Gene-specific effects of inflammatory cytokines on cytochrome P450 2C, 2B6 and 3A4 mRNA levels in human hepatocytes. Drug Metab Dispos. 2007;35:1687–93.

    Article  PubMed  CAS  Google Scholar 

  40. Gu X, Ke S, Liu D, Sheng T, Thomas PE, Rabson AB, et al. Role of NF-kappaB in regulation of PXR-mediated gene expression: a mechanism for the suppression of cytochrome P-450 3A4 by proinflammatory agents. J Biol Chem. 2006;281:17882–9.

    Article  PubMed  CAS  Google Scholar 

  41. Aitken AE, Lee CM, Morgan ET. Roles of nitric oxide in inflammatory downregulation of human cytochromes P450. Free Radic Biol Med. 2008;44:1161–8.

    Article  PubMed  CAS  Google Scholar 

  42. Pondugula SR, Dong H, Chen T. Phosphorylation and protein-protein interactions in PXR-mediated CYP3A repression. Expert Opin Drug Metab Toxicol. 2009;5:861–73.

    Article  PubMed  CAS  Google Scholar 

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ACKNOWLEDGEMENTS

The authors thank Dr. Steve Kliewer (University of Texas, Southwestern Medical Center, Dallas, TX) for providing the pSG5-hPXR expression vector. Human liver tissues were procured with the assistance of Jennifer Fuhrman from the University of Maryland Medical Center (Baltimore, MD). This research was partly supported by National Institute of Health Grant (R01, DK061652).

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Authors and Affiliations

  1. Department of Pharmaceutical Sciences, University of Maryland School of Pharmacy, 20 Penn Street, Baltimore, Maryland, 21201, USA

    Leslie Tompkins, Caitlin Lynch, James Polli & Hongbing Wang

  2. Food and Drug Administration, 7519 Metropark North, Rm. 3015A, HFD-600, Rockville, Maryland, 20855, USA

    Sam Haidar

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  1. Leslie Tompkins
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Correspondence to Hongbing Wang.

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Tompkins, L., Lynch, C., Haidar, S. et al. Effects of Commonly Used Excipients on the Expression of CYP3A4 in Colon and Liver Cells. Pharm Res 27, 1703–1712 (2010). https://doi.org/10.1007/s11095-010-0170-2

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  • DOI: https://doi.org/10.1007/s11095-010-0170-2

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