Original Articles
Yan Wen, Ruiqiang Zhao, Pranav Gupta, Yingfang Fan, Yunkai Zhang, Zhenguang Huang, Xiaohui Li, Yuangang Su, Lijuan Liao, Yu-An Xie, Donghua Yang, Zhe-Sheng Chen, Gang Liang. The epigallocatechin gallate derivative Y6 reverses drug resistance mediated by the ABCB1 transporter both in vitro and in vivo[J]. Acta Pharmaceutica Sinica B, 2019, 9(2): 316-323

The epigallocatechin gallate derivative Y6 reverses drug resistance mediated by the ABCB1 transporter both in vitro and in vivo
Yan Wena,b,c, Ruiqiang Zhaoa,d, Pranav Guptaa, Yingfang Fana,e, Yunkai Zhanga, Zhenguang Huangb, Xiaohui Lif, Yuangang Suf, Lijuan Liaof, Yu-An Xieg, Donghua Yanga, Zhe-Sheng Chena, Gang Liangh
a Department of Pharmaceutical Sciences, College of Pharmacy and Health Sciences, St. John's University, Queens, NY 11439, USA;
b Department of Pharmacy, the First Affiliated Hospital of Guangxi Medical University, Nanning 530021, China;
c Guangxi Colleges and Universities Key Laboratory of Biological Molecular Medicine Research, Guangxi Medical University, Nanning 530021, China;
d Department of Biochemistry and Molecular Biology, School of Preclinical Medicine, Guangxi Medical University, Nanning 530021, China e Department of Hepatobiliary Surgery, Zhujiang Hospital, Southern Medical University, Guangzhou 510282, China;
f Department of Biotechnology, School of Preclinical Medicine, Guangxi Medical University, Nanning 530021, China;
g The Affiliated Tumor Hospital of Guangxi Medical University, Nanning 530021, China;
h College of Pharmacy, Guangxi Medical University, Nanning 530021, China
Previously, we reported that Y6, a new epigallocatechin gallate derivative, is efficacious in reversing doxorubicin (DOX)-mediated resistance in hepatocellular carcinoma BEL-7404/DOX cells. In this study, we evaluated the efficacy of Y6 in reversing drug resistance both in vitro and in vivo by determining its effect on the adenosine triphosphate-binding cassette protein B1 transporter (ABCB1 or P-glycoprotein, P-gp). Our results showed that Y6 significantly sensitized cells overexpressing the ABCB1 transporter to anticancer drugs that are ABCB1 substrates. Y6 significantly stimulated the adenosine triphosphatase activity of ABCB1. Furthermore, Y6 exhibited a higher docking score as compared with epigallocatechin gallate inside the transmembrane domain of ABCB1. In addition, in the nude mouse tumor xenograft model, Y6 (110 mg/kg, intragastric administration), in combination with doxorubicin (2 mg/kg, intraperitoneal injection), significantly inhibited the growth of BEL-7404/DOX cell xenograft tumors, compared to equivalent epigallocatechin gallate. In conclusion, Y6 significantly reversed ABCB1-mediated multidrug resistance and its mechanisms of action may result from its competitive inhibition of the ABCB1 drug efflux function.
Key words:    Epigallocatechin gallate (EGCG)    5,3',4',3″,4″,5″-6-O-ethylEGCG (Y6)    Drug resistance    Resistance reversal    ABCB1    P-gp   
Received: 2018-06-21     Revised: 2018-08-16
DOI: 10.1016/j.apsb.2018.10.001
Funds: The authors would like to acknowledge Dr.Xiaocong Kuang (College of Basic Medicine,Guangxi Medical University,Nanning,China) for providing the BEL-7404/DOX cells.We thank Dr.Stephen Aller (The University of Alabama at Birmingham,Birmingham,US) for kindly providing human ABCB1 homology model.We thank Dr.Tanaji T.Talele (St.John's University,New York,NY,US) for providing the computational resources for molecular modeling,and Drs.Charles R.Ashby Jr.(St.John's University,Queens,NY,US) and Yangmin Chen (MediMedia Managed Markets,an ICON company,Yardley,PA,US) for reviewing and editing the paper.This work was supported by the National Natural Science Foundation of China (No.81160532),the Open Project of Guangxi Colleges and Universities Key Laboratory of Biological Molecular Medicine Research (No.GXBMR201602,China),the Young and Middle-aged Teachers Foundation Ability Enhancement Project of Guangxi Colleges and Universities (No.2018KY0102,China),and US NIH (No.1R15CA143701).
Corresponding author: Zhe-Sheng Chena, Gang Liang     Email:chenz@stjohns.edu;lianggang22@aliyun.com
Author description:
PDF(KB) Free
Yan Wen
Ruiqiang Zhao
Pranav Gupta
Yingfang Fan
Yunkai Zhang
Zhenguang Huang
Xiaohui Li
Yuangang Su
Lijuan Liao
Yu-An Xie
Donghua Yang
Zhe-Sheng Chen
Gang Liang

1. Gottesman MM, Fojo T, Bates SE. Multidrug resistance in cancer. Role of ATP-dependent transporters. Nat Rev Cancer 2002;92:48-58.
2. Sodani K, Patel A, Kathawala RJ, Chen ZS. Multidrug resistance associated proteins in multidrug resistance. Chin J Cancer 2012;31:58-72.
3. Gottesman MM. Mechanisms of cancer drug resistance. Ann Rev Med 2002;53:615-27.
4. Fan YF, Zhang W, Zeng L, Lei ZN, Cai CY, Gupta P, et al. Dacomitinib antagonizes multidrug resistance (MDR) in cancer cells by inhibiting the efflux activity of ABCB1 and ABCG2 transporters. Cancer Lett 2018;421:186-98.
5. An X, Sarmiento C, Tan T, Zhu H. Regulation of multidrug resistance by microRNAs in anti-cancer therapy. Acta Pharm Sin B 2017;7:38-51.
6. Borths EL, Locher KP, Lee AT, Rees DC. The structure of Escherichia coli BtuF and binding to its cognate ATP binding cassette transporter. Proc Nati Acad Sci U S A 2002;99:16642-7.
7. Locher KP. Structure and mechanism of ABC transporters. Curr Opin Struct Biol 2004;14:426-31.
8. Locher KP, Borths E. ABC transporter architecture and mechanism:implications from the crystal structures of BtuCD and BtuF. FEBS Lett 2004;564:264-8.
9. Dean M, Allikmets R. Complete characterization of the human ABC gene family. J Bioenerg Biomembr 2001;33:475-9.
10. Dean M, Annilo T. Evolution of the ATP-binding cassette (ABC) transporter superfamily in vertebrates. Annu Rev Genom Hum Genet 2005;6:123-42.
11. Schinkel AH, Jonker JW. Mammalian drug efflux transporters of the ATP binding cassette (ABC) family:an overview. Adv Drug Deliv Rev 2003;55:3-29.
12. Juliano RL, Ling V. A surface glycoprotein modulating drug permeability in Chinese hamster ovary cell mutants. Biochim Biophys Acta 1976;455:152-62.
13. Ueda K, Cornwell MM, Gottesman MM, Pastan I, Roninson IB, Ling V, et al. The mdr1 gene, responsible for multidrug-resistance, codes for P-glycoprotein. Biochem Biophys Res Commun 1986;141:956-62.
14. Tiwari AK, Sodani K, Dai CL, Ashby Jr CR, Chen ZS. Revisiting the ABCs of multidrug resistance in cancer chemotherapy. Curr Pharm Biotechnol 2011;12:570-94.
15. Wu CP, Hsieh CH, Wu YS. The emergence of drug transportermediated multidrug resistance to cancer chemotherapy. Mol Pharm 2011;8:1996-2011.
16. Roninson IB, Chin JE, Choi KG, Gros P, Housman DE, Fojo A, et al. Isolation of human mdr DNA sequences amplified in multidrug-resistant KB carcinoma cells. Proc Natl Acad Sci U S A 1986;83:4538-42.
17. Van der Bliek AM, Baas F, Ten Houte de Lange T, Kooiman PM, van der Velde-Koerts T, Borst P. The human mdr3 gene encodes a novel Pglycoprotein homologue and gives rise to alternatively spliced mRNAs in liver. EMBO J 1987;6:3325-31.
18. de Bruijn MH, Van der Bliek AM, Biedler JL, Borst P. Differential amplification and disproportionate expression of five genes in three multidrug-resistant Chinese hamster lung cell lines. Mol Cell Biol 1986;6:4717-22.
19. Gros P, Raymond M, Bell J, Housman D. Cloning and characterization of a second member of the mouse mdr gene family. Mol Cell Biol 1988;8:2770-8.
20. Aller SG, Yu J, Ward A, Weng Y, Chittaboina S, Zhuo R, et al. Structure of P-glycoprotein reveals a molecular basis for poly-specific drug binding. Science 2009;323:1718-22.
21. Wu CP, Calcagno AM, Ambudkar SV. Reversal of ABC drug transporter-mediated multidrug resistance in cancer cells:evaluation of current strategies. Curr Mol Pharmacol 2008;1:93-105.
22. Wen Y, Zhao RQ, Zhang YK, Gupta P, Fu LX, Tang AZ, et al. Effect of Y6, an epigallocatechingallate derivative, on reversing doxorubicin drug resistance in human hepatocellular carcinoma cells. Oncotarget 2017;8:29760-70.
23. Liang G, Tang A, Lin X, Li L, Zhang S, Huang Z, et al. Green tea catechins augment the antitumor activity of doxorubicin in an in vivo mouse model for chemoresistant liver cancer. Int J Oncol 2010;37:111-23.
24. Kitagawa S, Nabekura T, Kamiyama S. Inhibition of P-glycoprotein function by tea catechins in KB-C2 cells. J Pharm Pharmacol 2004;56:1001-5.
25. Zhang H, Zhang YK, Wang YJ, Kathawala RJ, Patel A, Zhu H, et al. WHI-P154 enhances the chemotherapeutic effect of anticancer agents in ABCG2-overexpressing cells. Cancer Sci 2014;105:1071-8.
26. Li J, Jaimes KF, Aller SG. Refined structures of mouse P-glycoprotein. Protein Sci 2014;23:34-46.
27. Zhang YK, Zhang GN, Wang YJ, Patel BA, Talele TT, Yang DH, et al. Bafetinib (INNO-406) reverses multidrug resistance by inhibiting the efflux function of ABCB1 and ABCG2 transporters. Sci Rep 2016;6:25694.
28. Zhang YK, Zhang H, Zhang GN, Wang YJ, Kathawala RJ, Si R, et al. Semi-synthetic ocotillol analogues as selective ABCB1-mediated drug resistance reversal agents. Oncotarget 2015;6:24277-90.
29. Jain AN. Surflex:fully automatic flexible molecular docking using a molecular similarity-based search engine. J Med Chem 2003;46:499-511.
30. Wang YJ, Kathawala RJ, Zhang YK, Patel A, Kumar P, Shukla S, et al. Motesanib (AMG706), a potent multikinase inhibitor, antagonizes multidrug resistance by inhibiting the efflux activity of the ABCB1. Biochem Pharmacol 2014;90:367-78.
31. Sauna ZE, Ambudkar SV. Evidence for a requirement for ATP hydrolysis at two distinct steps during a single turnover of the catalytic cycle of human P-glycoprotein. Proc Natl Acad Sci U S A 2000;97:2515-20.
32. Sauna ZE, Ambudkar SV. Characterization of the catalytic cycle of ATP hydrolysis by human P-glycoprotein the two ATP hydrolysis events in a single catalytic cycle are kinetically similar but affect different functional outcomes. J Biol Chem 2001;276:11653-61.
33. Ambudkar SV, Dey S, Hrycyna CA, Ramachandra M, Pastan I, Gottesman MM. Biochemical, cellular, and pharmacological aspects of the multidrug transporter 1. Annu Rev Pharmacol Toxicol 1999;39:361-98.
34. Pajeva IK, Globisch C, Wiese M. Combined pharmacophore modeling, docking, and 3D QSAR studies of ABCB1 and ABCC1 transporter inhibitors. Chem Med Chem 2009;4:1883-96.