CYTOTOXIC ACTIVITY OF BRAZILEIN ISOLATED FROM SECANG (CAESALPINIA SAPPAN L.) AGAINST MCF7/DOX CELLS BY INHIBITION OF P-GLYCOPROTEIN
Objective: This study was focused on isolation of brazilein from the dried heartwood of Secang (Caesalpinia sappan L.) followed by its characterization using Infrared (IR) spectroscopy, liquid chromatography-mass spectrometry (LC-MS) with electrospray ionization (ESI), proton (1H), carbon-13 (13C) nuclear magnetic resonance (NMR) and two dimensional (2D)-NMR, evaluation the cytotoxic activity of brazilein in MCF-7 resistant doxorubicin (MCF-7/DOX) cells and evaluate the interaction between brazilein and ATP with P-glycoprotein (Pgp) in silico using molecular docking.
Methods: Brazilein was isolated and purified from ethyl acetate fraction by flash silica gel column chromatography, eluting with chloroform, ethyl acetate and methanol in gradient concentration. In the cytotoxicity assay, MCF-7/DOX cells were cultured in the presence of brazilein for 24 hour (h) and cell viability was evaluated by using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay. Interactions between brazilein and the target proteins were evaluated and calculated in silico by molecular docking using PLANTS.
Results: The infrared, mass spectra with a molecular weight of 284 and NMR signal confirmed that was brazilein. MTT assay showed a dose-dependent inhibition of MCF-7/DOX cell proliferation with brazilein IC50 value of 43 ÂµM. The docking score of brazilein was-71,45 kcal/mol and ATP value-96,23 kcal/mol.
Conclusion: Brazilein has a potent cytotoxic value on MCF-7/DOX and high affinity in Pgp protein target. Brazilein can be developed as anticancer especially in cancer resistance incidence. Further study must be established to evaluate the molecular mechanism of brazilein inhibit MCF-7/DOX cell proliferation in vitro.
2. Ye M, Xie WD, Lei F, Meng Z, Zhao YN, Su H, et al. Brazilein, an important immunosuppressive component from Caesalpinia sappan L. Int J Immunopharmacol 2006;6:426-32.
3. Hu J, Yan X, Wang W, Wu H, Hua L, Du L. Antioxidant activity in vitro of three constituents from Caesalpinia sappan L. Tsinghua Sci Technol 2008;13:474-9.
4. Lim MY, Jeon JH, Jeong EY, Lee CH, Lee HS. Antimicrobial activity of 5-hydroxy-1,4-naphthoquinone isolated from Caesalpinia sappan toward intestinal bacteria. Food Chem 2007;100:1254-8.
5. Batubara I, Mitsunaga T, Ohashi H. Brazilin from Caesalpinia sappan wood as an antiacne agent. J Wood Sci 2009;56:77-81.
6. Zhong B, Wu YJ Pan, S Zheng. Brazilein inhibits survivin protein and mRNA expression and induces apoptosis in hepatocellular carcinoma HepG2 cells. Neoplasma 2009;56:387-92.
7. Lim DK, U Choi, DH Shin. Antioxidative activity of some solvent extract from Caesalpinia sappan Linn, Korean J. Food Sci Technol 1997;28:77âˆ’82.
8. Yen C, Kyoko Nakagawa-Goto, Tsong-Long Hwang, Pei-Chi Wu, Susan L Morris-Natschke, Wan-Chun Lai, et al. Antitumor agents. 271. total synthesis and evaluation of brazilein and analogs as anti-inflammatory and cytotoxic agents. Bioorg Med Chem Lett 2010;20:1037â€“9.
9. Tao LY, Li JY, Zhang JY. Brazilein, a compound isolated from Caesalpinia sappan Linn., induced growth inhibition in breast cancer cells via involvement of GSK-3ï¢/ï¢-Catenin/cyclin D1 pathway. Chem Biol Interact 2013;206:1-5.
10. Laksmiani NPL, Susidarti RA, Meiyanto E. Brazilein increases the sensitivity of doxorubicin on MCF-7 resistant doxorubicin (MCF-7/DOX) cells through inhibition of HER-2 activation. Int J Pharm Pharm Sci 2015;7:525-8.
11. Byun SS, Kim SW, Choi H, Lee C, Lee E. Augmentation of cisplatin sensitivity in cisplatin-resistant human bladder cancer cells by modulating glutathione concentrations and glutathione-related enzyme activities. BJU Int 2005;95: 1086-90.
12. Stavrovskaya AA, Stromskaya TP. Transport proteins of the ABC family and multidrug resistance of tumor cells. Biochemistry (Moscow) 2008;73 :592-604.
13. Chung SY, Sung MK, Kim NH, Jang JO, Go EJ, Lee HJ. Inhibition of P-glycoprotein by natural products in human breast cancer cells. Arch Pharm Res 2005;28:823-8.
14. Wong HL, Bendayan R, Rauth AM, Xue HY, Babakhanian K, Wu XY. A mechanistic study of enhanced doxorubicin uptake and retention in multidrug resistant breast cancer cells using a polymer-lipid hybrid nanoparticle system. J Pharmacol Exp Ther 2006;317:1372-81.
15. Putri DDP, Sarmoko, Febriansah R. MCF-7 resistant doxorubicin are characterized by lamellipodia, strong adhesion on the substrate and Pgp overexpression. Indo J Can Chemoprev 2013;2:304-8.
16. Skoog DA, Holler FJ, Crouch SR. Principles of instrumental analysis. Sixth Edition. Thomson Higher Education, Canada USA; 2007.
17. Pavia DL, Lampman GM, Kriz GS. Introduction to spectroscopy. 2nd ed. Philadelphia: Harcourt Brace College Publ; 1996. p. 511.
18. Oliveira LFC, Edwards HGM, Velozo ES, Nesbitt M. Vibrational spectroscopic study of Brazilin and Brazilein, the main constituents of brazilwood from Brazil. Vibrational Spectroscopy 2002;28:243-9.
19. Kim DS, Baek NI, Jung KY, Lee IS, Lee HK. NMR assignment of Brazilein. Phytochemistry 1997;46:177-8.
20. Rosenberg E. Characterisation of historical organic dyestuffs by liquid chromatography-mass spectrometry. Anal Bioanal Chem 2008;391:33-57.
21. Gajjar DG, Patel RM, Patel VA, Patel PKM. Novel hydroxyl terminated dendrimers as potential drug carriers: sustained release, hemolysis and cytotoxicity study. Int J Appl Pharm 2015;7:5-9.
22. Teng WY, Yu LH, Chien CS, Ray LH. Cytotoxic acridone alkaloids from te stem bark of citrus maxima. J Chin Chem Soc 2005;52:1253-5.
23. Sanneboina S, Sammeta V, Pingili RB, Kadimpati KK. Influence of lovastatin on pharmacokinetics and pharmacodynamics of glipizide in healthy and streptozotocin-induced diabetic rats: involvement of p-glycoprotein inhibition. Asian J Pharm Clin Res 2016;9:3-8.
24. Aller SG, Yu J, Ward A, Weng Y, Chittaboina S, Zhou R, et al. Structure of P-glycoprotein reveals a molecular basis for poly-specific drug binding. Science 2012;323:171-2.