Different structures of berberine and five other protoberberine alkaloids that affect P-glycoprotein-mediated efflux capacity

Article metrics

  • 403 Accesses

  • 1 Citations

Abstract

Berberine, berberrubine, thalifendine, demethyleneberberine, jatrorrhizine, and columbamine are six natural protoberberine alkaloid (PA) compounds that display extensive pharmacological properties and share the same protoberberine molecular skeleton with only slight substitution differences. The oral delivery of most PAs is hindered by their poor bioavailability, which is largely caused by P-glycoprotein (P-gp)-mediated drug efflux. Meanwhile, P-gp undergoes large-scale conformational changes (from an inward-facing to an outward-facing state) when transporting substrates, and these changes might strongly affect the P-gp-binding specificity. To confirm whether these six compounds are substrates of P-gp, to investigate the differences in efflux capacity caused by their trivial structural differences and to reveal the key to increasing their binding affinity to P-gp, we conducted a series of in vivo, in vitro, and in silico assays. Here, we first confirmed that all six compounds were substrates of P-gp by comparing the drug concentrations in wild-type and P-gp-knockout mice in vivo. The efflux capacity (net efflux) ranked as berberrubine > berberine > columbamine ~ jatrorrhizine > thalifendine > demethyleneberberine based on in vitro transport studies in Caco-2 monolayers. Using molecular dynamics simulation and molecular docking techniques, we determined the transport pathways of the six compounds and their binding affinities to P-gp. The results suggested that at the early binding stage, different hydrophobic and electrostatic interactions collectively differentiate the binding affinities of the compounds to P-gp, whereas electrostatic interactions are the main determinant at the late release stage. In addition to hydrophobic interactions, hydrogen bonds play an important role in discriminating the binding affinities.

Access options

Rent or Buy article

Get time limited or full article access on ReadCube.

from$8.99

All prices are NET prices.

Scheme 1
Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6

References

  1. 1.

    Preininger V. Chemotaxonomy of papaveraceae and fumariaceae. In: Arnold B, editor. The alkaloids: chemistry and pharmacology. vol. 29. New York: Harcourt Brace Jovanovich; 1986. p. 1–98.

  2. 2.

    Kumar A, Ekavali, Chopra K, Mukherjee M, Pottabathini R, Dhull DK. Current knowledge and pharmacological profile of berberine: an update. Eur J Pharmacol. 2015;761:288–97.

  3. 3.

    Bandyopadhyay S, Patra PH, Mahanti A, Mondal DK, Dandapat P, Bandyopadhyay S, et al. Potential antibacterial activity of berberine against multi drug resistant enterovirulent Escherichia coli isolated from yaks (Poephagus grunniens) with haemorrhagic diarrhoea. Asian Pacific J Trop Med. 2013;6:315–9.

  4. 4.

    Zhang Q, Xiao XH, Feng K, Wang T, Li WH, Yuan T, et al. Berberine moderates glucose and lipid metabolism through multipathway mechanism. Evid Based Complement Alternat Med. 2011; 1–10.

  5. 5.

    Chang XX, Yan HM, Xu Q, Xia MF, Bian H, Zhu TF, et al. The effects of berberine on hyperhomocysteinemia and hyperlipidemia in rats fed with a long-term high-fat diet. Lipids Health Dis. 2012;11:86.

  6. 6.

    Wang LH, Liu LP, Shi Y, Cao HW, Chaturvedi R, Calcutt MW, et al. Berberine induces caspase-independent cell death in colon tumor cells through activation of apoptosis-inducing factor. PLoS ONE. 2012;7:e36418.

  7. 7.

    Rabbani GH, Butler T, Knight J, Sanyal SC, Alam K. Randomized controlled trial of berberine sulfate therapy for diarrhea due to enterotoxigenic Escherichia coli and Vibrio cholerae. J Infect Dis. 1987;155:979–84.

  8. 8.

    Iranshahy M, Quinn RJ, Iranshahi M. Biologically active isoquinoline alkaloids with drug-like properties from the genus Corydalis. RSC Adv. 2014;4:15900–13.

  9. 9.

    Bhadra K, Kumar GS. Therapeutic potential of nucleic acid-binding isoquinoline alkaloids: binding aspects and implications for drug design. Med Res Rev. 2011;31:821–62.

  10. 10.

    Grycova L, Dostal J, Marek R. Quaternary protoberberine alkaloids. Phytochemistry. 2007;68:150–75.

  11. 11.

    Parcej D, Tampe R. ABC proteins in antigen translocation and viral inhibition. Nat Chem Biol. 2010;6:572–80.

  12. 12.

    Thiebaut F, Tsuruo T, Hamada H, Gottesman MM, Pastan I, Willingham MC. Cellular-localization of the multidrug-resistance gene-product P-glycoprotein in normal human-tissues. Proc Natl Acad Sci USA. 1987;84:7735–8.

  13. 13.

    Borst P, Schinkel AH. P-glycoprotein ABCB1: a major player in drug handling by mammals. J Clin Invest. 2013;123:4131–3.

  14. 14.

    Liu CS, Zheng YR, Zhang YF, Long XY. Research progress on berberine with a special focus on its oral bioavailability. Fitoterapia. 2016;109:274–82.

  15. 15.

    Sun YH, He X, Yang XL, Dong CL, Zhang CF, Song ZJ, et al. Absorption characteristics of the total alkaloids from Mahonia bealei in an in situ single-pass intestinal perfusion assay. Chin J Nat Med. 2014;12:554–60.

  16. 16.

    Chen CP, Liu XR, Smith BJ. Utility of mdr1-gene deficient mice in assessing the impact of P-glycoprotein on pharmacokinetics and pharmacodynamics in drug discovery and development. Curr Drug Metab. 2003;4:272–91.

  17. 17.

    Collett A, Tanianis-Hughes J, Hallifax D, Warhurst G. Predicting P-glycoprotein effects on oral absorption: correlation of transport in Caco-2 with drug pharmacokinetics in wild-type and mdr1a(−/−) mice in vivo. Pharm Res. 2004;21:819–26.

  18. 18.

    Aller SG, Yu J, Ward A, Weng Y, Chittaboina S, Zhuo RP, et al. Structure of P-glycoprotein reveals a molecular basis for poly-specific drug binding. Science. 2009;323:1718–22.

  19. 19.

    Pajeva IK, Globisch C, Wiese M. Combined pharmacophore modeling, docking, and 3D QSAR studies of ABCB1 and ABCC1 transporter inhibitors. ChemMedChem. 2009;4:1883–96.

  20. 20.

    Dolghih E, Bryant C, Renslo AR, Jacobson MP. Predicting binding to p-glycoprotein by flexible receptor docking. PLoS Comput Biol. 2011;7:e1002083.

  21. 21.

    Smith PC, Karpowich N, Millen L, Moody JE, Rosen J, Thomas PJ, et al. ATP binding to the motor domain from an ABC transporter drives formation of a nucleotide sandwich dimer. Mol Cell. 2002;10:139–49.

  22. 22.

    Verhalen B, Wilkens S. P-glycoprotein retains drug-stimulated ATPase activity upon covalent linkage of the two nucleotide binding domains at their C-terminal ends. J Biol Chem. 2011;286:10476–82.

  23. 23.

    Subramanian N, Condic-Jurkic K, O’Mara ML. Structural and dynamic perspectives on the promiscuous transport activity of P-glycoprotein. Neurochem Int. 2016;98:146–52.

  24. 24.

    Qiu F, Zhu Z, Kang N, Piao S, Qin G, Yao X. Isolation and identification of urinary metabolites of berberine in rats and humans. Drug Metab Dispos. 2008;36:2159–65.

  25. 25.

    Ma JY, Feng R, Tan XS, Ma C, Shou JW, Fu J, et al. Excretion of berberine and its metabolites in oral administration in rats. J Pharm Sci. 2013;102:4181–92.

  26. 26.

    Ma BL, Ma YM. Pharmacokinetic properties, potential herb-drug interactions and acute toxicity of oral Rhizoma coptidis alkaloids. Expert Opin Drug Metab Toxicol. 2013;9:51–61.

  27. 27.

    Li Y, Ren G, Wang YX, Kong WJ, Yang P, Wang YM, et al. Bioactivities of berberine metabolites after transformation through CYP450 isoenzymes. J Transl Med. 2011;9:62.

  28. 28.

    Zhang X, Qiu F, Jiang J, Gao C, Tan Y. Intestinal absorption mechanisms of berberine, palmatine, jateorhizine, and coptisine: involvement of P-glycoprotein. Xenobiotica. 2011;41:290–6.

  29. 29.

    Li J, Jaimes KF, Aller SG. Refined structures of mouse P-glycoprotein. Protein Sci. 2014;23:34–46.

  30. 30.

    Pan L, Aller SG. Equilibrated atomic models of outward-facing P-glycoprotein and effect of ATP binding on structural dynamics. Sci Rep. 2015;5:7880.

  31. 31.

    Li MJ, Nath A, Atkins WM. Differential coupling of binding, ATP hydrolysis, and transport of fluorescent probes with P-glycoprotein in lipid nanodiscs. Biochemistry. 2017;56:2506–17.

  32. 32.

    Frank GA, Shukla S, Rao P, Borgnia MJ, Bartesaghi A, Merk A, et al. Cryo-EM analysis of the conformational landscape of human P-glycoprotein (ABCB1) during its catalytic cycle. Mol Pharmacol. 2016;90:35–41.

  33. 33.

    Wise JG. Catalytic transitions in the human MDR1 P-glycoprotein drug binding sites. Biochemistry. 2012;51:5125–41.

  34. 34.

    Ferreira RJ, Ferreira M-JU, dos Santos DJVA. Insights on P-glycoprotein’s efflux mechanism obtained by molecular dynamics simulations. J Chem Theory Comput. 2012;8:1853–64.

  35. 35.

    McCormick JW, Vogel PD, Wise JG. Multiple drug transport pathways through human P-glycoprotein. Biochemistry. 2015;54:4374–90.

  36. 36.

    Wang J, Shao Q, Xu Z, Liu Y, Yang Z, Cossins BP, et al. Exploring transition pathway and free-energy profile of large-scale protein conformational change by combining normal mode analysis and umbrella sampling molecular dynamics. J Phys Chem B. 2014;118:134–43.

  37. 37.

    Volpe DA, Faustino PJ, Ciavarella AB, Asafu-Adjaye EB, Ellison CD, Yu LX, et al. Classification of drug permeability with a Caco-2 cell monolayer assay. Clin Res Regul Aff. 2007;24:39–47.

  38. 38.

    Chufan EE, Kapoor K, Ambudkar SV. Drug-protein hydrogen bonds govern the inhibition of the ATP hydrolysis of the multidrug transporter P-glycoprotein. Biochem Pharmacol. 2016;101:40–53.

  39. 39.

    Lin JH. Drug–drug interaction mediated by inhibition and induction of P-glycoprotein. Adv Drug Deliv Rev. 2003;55:53–81.

  40. 40.

    Zhang Y, Bachmeier C, Miller DW. In vitro and in vivo models for assessing drug efflux transporter activity. Adv Drug Deliv Rev. 2003;55:31–51.

  41. 41.

    Schinkel AH, Mayer U, Wagenaar E, Mol CA, van Deemter L, Smit JJ, et al. Normal viability and altered pharmacokinetics in mice lacking mdr1-type (drug-transporting) P-glycoproteins. Proc Natl Acad Sci USA. 1997;94:4028–33.

  42. 42.

    Volpe DA. Drug-permeability and transporter assays in Caco-2 and MDCK cell lines. Fut Med Chem. 2011;3:2063–77.

  43. 43.

    Polli JW, Wring SA, Humphreys JE, Huang LY, Morgan JB, Webster LO, et al. Rational use of in vitro P-glycoprotein assays in drug discovery. J Pharmacol Exp Ther. 2001;299:620–8.

  44. 44.

    Wang Q, Strab R, Kardos P, Ferguson C, Li J, Owen A, et al. Application and limitation of inhibitors in drug–transporter interactions studies. Int J Pharm. 2008;356:12–8.

  45. 45.

    Lennernas H. Intestinal permeability and its relevance for absorption and elimination. Xenobiotica. 2007;37:1015–51.

  46. 46.

    Ferreira RJ, Ferreira M-JU, dos Santos DJVA. Do drugs have access to the P-glycoprotein drug-binding pocket through gates? J Chem Theory Comput. 2015;11:4525–9.

  47. 47.

    Bansal T, Akhtar N, Jaggi M, Khar RK, Talegaonkar S. Novel formulation approaches for optimising delivery of anticancer drugs based on P-glycoprotein modulation. Drug Discov Today. 2009;14:1067–74.

  48. 48.

    Malingre MM, Beijnen JH, Rosing H, Koopman FJ, van Tellingen O, Duchin K, et al. A phase I and pharmacokinetic study of bi-daily dosing of oral paclitaxel in combination with cyclosporin A. Cancer Chemother Pharmacol. 2001;47:347–54.

  49. 49.

    Malingre MM, Richel DJ, Beijnen JH, Rosing H, Koopman FJ, Ten Bokkel Huinink WW, et al. Coadministration of cyclosporine strongly enhances the oral bioavailability of docetaxel. J Clin Oncol. 2001;19:1160–6.

  50. 50.

    Shan YQ, Ren G, Wang YX, Pang J, Zhao ZY, Yao J, et al. Berberine analogue IMB-Y53 improves glucose-lowering efficacy by averting cellular efflux especially P-glycoprotein efflux. Metabolism. 2013;62:446–56.

  51. 51.

    Li YH, Yang P, Kong WJ, Wang YX, Hu CQ, Zuo ZY, et al. Berberine analogues as a novel class of the low-density-lipoprotein receptor up-regulators: synthesis, structure–activity relationships, and cholesterol-lowering efficacy. J Med Chem. 2009;52:492–501.

  52. 52.

    Cui HM, Zhang QY, Wang JL, Chen JL, Zhang YL, Tong XL. Poor permeability and absorption affect the activity of four alkaloids from Coptis. Mol Med Rep. 2015;12:7160–8.

Download references

Acknowledgements

This work was partly supported by the National Natural Science Foundation of China (Grants 81573499 81872927) and Strategic Priority Research Program of the Chinese Academy of Sciences (Grant XDA16020205). We thank Prof. Cheng-gang Huang (Shanghai Institute of Materia Medica, Chinese Academy of Sciences) for providing the purified compound.

Author contributions

Y-tZ, Y-qY, X-tT, W-lZ, L-kG, and G-yP were responsible for the research design and wrote this manuscript; Y-tZ, Y-qY, X-xY, LW, and W-jW conducted the experiments; Y-qY and W-lZ contributed analytical tools; and Y-tZ and Y-qY performed the data analysis.

Author information

Correspondence to Wei-liang Zhu or Li-kun Gong or Guo-yu Pan.

Ethics declarations

Competing interests

The authors declare no competing interests.

Electronic supplementary material

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Zhang, Y., Yu, Y., Yan, X. et al. Different structures of berberine and five other protoberberine alkaloids that affect P-glycoprotein-mediated efflux capacity. Acta Pharmacol Sin 40, 133–142 (2019) doi:10.1038/s41401-018-0183-7

Download citation

Keywords

  • P-glycoprotein
  • protoberberine alkaloid
  • berberine
  • binding affinity
  • efflux

Further reading