Hydrazinyldiene-chroman-2,4-diones in inducing growth arrest and apoptosis in breast cancer cells: Synergism with doxorubicin and correlation with physicochemical properties

Open access


This study evaluates the effects of previously synthesized hydrazinyldiene-chroman-2,4-diones on cell proliferation and apoptosis, cell cycle distribution and migration capacity of MCF-7 breast cancer cells in synergy with doxorubicin. Physicochemical properties of the synthesized compounds were correlated with their structure and activity. Significant cell viability decrease in comparison with the effect of doxorubicin alone and the reference 4-hydroxycoumarin was observed when combination treatment comprising doxorubicin and the title compounds was applied. Synergistic effect with doxorubicin was also observed in down-regulation of phospho-Thr308Akt levels, confirming reduced proliferation and increased apoptosis. Combined treatment increased the percentage of cells arrested at the G2/M stage. Additive inhibition of cell migration was also observed, pointing to the possibility of reducing the risk of metastases. With their solubility profile and log D7.4, all the synthesized compounds follow Lipinski’s rule of five for good permeability (absorption) potential.

1. A. M. Stuckey, Breast cancer: epidemiology and risk factors, Clin. Obst. Gynecol. 54 (2011) 96-102; DOI: 10.1097/GRF.0b013e3182080056.

2. F. C. Thorn, C. Oshiro, S. Marsh, T. Hernandez-Boussard, H. McLeod, T. Klein and B. R. Altman, Doxorubicin pathways: pharmacodynamics and adverse effects, Pharmacogenet. Genom. 21 (2011) 440-446; DOI: 10.1097/FPC.0b013e32833ffb56.

3. A. Jashari, F. Imeri, L. Ballazhi, A. Shabani, B. Mikhova, G. Dräger, E. Popovski and A. Huwiler, Synthesis and cellular characterization of novel isoxazolo- and thiazolohydrazinyldiene-chroman-2,4-diones on cancer and non-cancer cell growth and death, Bioorg. Med. Chem. 22 (2014) 2655-2661; DOI: 10.1016/j.bmc.2014.03.026.

4. S. Chen, M. Cho, K. Karlsberg, D. Zhou and Y. Yuan, Biochemical and biological characterization of a novel anti-aromatase coumarin derivative, J. Biol. Chem. 279 (2004) 48071-48078; DOI: 10.1074/ jbc.M406847200.

5. L. Woo, A. Purohit, B. Malini, M. Reed and B. Potter, Potent active site-directed inhibition of steroid sulphatase by tricyclic coumarin-based sulphamates, Chem. Biol. 7 (2000) 773-791; DOI: 10.1016/S1074-5521(00)00023-5.

6. M. Musa, J. Cooperwood and M. Khan, A review of coumarin derivatives in pharmacotherapy of breast cancer, Curr. Med. Chem. 15 (2008) 2664-2679; DOI: 10.2174/092986708786242877.

7. S. Kirkiacharian, A. Lormier, H. Chidiack, F. Bouchoux and E. Cérède, Synthesis and binding affinity to human alpha and beta estrogen receptors of various 7-hydroxycoumarins substituted at 4- and 3,4- positions, Farmaco 59 (2004) 981-986; DOI: 10.1016/j.farmac.2004.08.004.

8. M. Halasi, H. Zhao, H. Dahari, U. G. Bhat, E. B. Gonzalez, A. V. Lyubimo, D. A. Tonetti and A. L. Gartel, Thiazole antibiotics against breast cancer, Cell Cycle 9 (2010) 1214-1217; DOI: 10.4161/ cc.9.6.10955.

9. J. Shaw, B. Chen, J. P. Bourgault, H. Jiang, K. Narendra, M. Jayshree, A. V. Frederick, M. Joe, B. Kevin, P. Halina, E. Matthew and R. A. Peter, Synthesis and biological evaluation of novel nphenyl-5-carboxamidyl isoxazoles as potential chemotherapeutic agents for colon cancer, Am. J. Biomed. Sci. 4 (2012) 14-25; DOI: 10.5099/aj120100014.

10. P. Poma, M. Notarbartolo, M. Labbozzetta, A. Maurici, V. Carina, A. Alaimo, M. Rizzi, D. Simoni and N. D’Alessandro, The antitumor activities of curcumin and of its isoxazole analogue are not affected by multiple gene expression changes in an MDR model of the MCF-7 breast cancer cell line: analysis of the possible molecular basis, Int. J. Mol. Med. 20 (2007) 329-335; DOI: 10.3892/ ijmm.20.3.329.

11. L. Ballazhi, E. Popovski, A. Jashari, F. Imeri, I. Ibrahimi, B. Mikhova and K. Mladenovska, Potential antiproliferative effect of isoxazolo- and thiazolo coumarin derivatives on breast cancer mediated bone and lung metastases, Acta Pharm. 65 (2015) 53-63; DOI: 10.1515/acph-2015-0002.

12. A. Jashari, E. Popovski, B. Mikhova, P. R. Nikolova and L. B. Shivachev, 3-[2-(5-tert-Butyl-1,2-oxazol-3-yl)hydrazinylidene]chroman-2,4-dione, Acta Crystallogr. E69 (2013) o258; DOI: 10.1107/ S1600536813000858.

13. L. Ballazhi, F. Imeri, A. Dimovski, A. Jashari, E. Popovski, P. B. Selmani, B. Mikhova, G. Dräger, E. Alili-Idrizi and K. Mladenovska, Synergy of novel coumarin derivatives and tamoxifen in blocking growth and inducing apoptosis of breast cancer cells, Mac. Pharm. Bull. 60 (2014) 35-44.

14. N. C. Bols, V. R. Dayeh, L. E. J. Lee and K. Schirmer, Use of Fish Cell Lines in the Toxicology and Ecotoxicology of Fish, in Biochemistry and Molecular Biology of Fishes - Environmental Toxicology (Eds. T. P. Mommsen and T. W. Moon), Vol. 6, Elsevier Science, Amsterdam 2005, pp. 43-84.

15. R. Hamid, Y. Rotshteyn, L. Rabadi, R. Parikh and P. Bullock, Comparison of alamar blue and MTT assays for high through-put screening, Toxicol. In Vitro 18 (2004) 703-710; DOI: 10.1016/j. tiv.2004.03.012.

16. W. F. Deckert and K. H. Remmer, In vitro inhibition of rat and human liver microsomal enzymes by 4-hydroxycoumarin anticoagulants and related compounds, Chem. Biol. Inter. 5 (1972) 255-263; DOI: 10.1016/0009-2797(72)90029-4.

17. H. Liu, Y. Wang, A. Sharma, R. Mao, N. Jiang, B. Dun and X. J. She, Derivatives containing both coumarin and benzimidazole potently induce caspase-dependent apoptosis of cancer cells through inhibition of PI3K-AKT-mTOR signaling, Anticancer Drugs 26 (2015) 667-677; DOI: 10.1097/ CAD.0000000000000232.

18. O. Bar-On, M. Shapira and D. D. Hershko, Differential effects of doxorubicin treatment on cell cycle arrest and Skp2 expression in breast cancer cells, Anti-Cancer Drugs 18 (2007) 1113-1121; DOI: 10.1097/CAD.0b013e3282ef4571.

19. R. Lüpertz, W. Wätjen, R. Kahl and Y. Chovolou, Dose- and time-dependent effects of doxorubicin on cytotoxicity, cell cycle and apoptotic cell death in human colon cancer cells, Toxicology 271 (2010) 115-121; DOI: 10.1016/j.tox.2010.03.012.

20. A. M. Musa, L. V. Badisa, M. L. Latinwo, J. Cooperwood, A. Sinclair and A. Abdullah, Cytotoxic activity of new acetoxycoumarin derivatives in cancer cell lines, Anticancer Res. 31 (2011) 2017-2022.

21. A. Goel, K. A. Prasad, S. V. Parmar, B. Ghosh and N. Saini, 7,8-Dihydroxy-4-methylcoumarin induces apoptosis of human lung adenocarcinoma cells by ROS-independent mitochondrial pathway through partial inhibition of ERK/MAPK signaling, FEBS Lett. 581 (2007) 2447-2454; DOI: 10.1016/j.febslet.2007.04.052.

22. D. Alesiani, R. Cicconi, M. Mattei, C. Montesano, R. Bei and A. Canini, Cell cycle arrest and differentiation induction by 5,7-dimethoxycoumarin in melanoma cell lines, Int. J. Oncol. 32 (2008) 425-434; DOI: 10.3892/ijo.32.2.425.

23. F. Haghighi, M. M. Matin, A. R. Bahrami, M. Iranshahi, F. B. Rassouli and A. Haghighitalab, The cytotoxic activities of 7-isopentenyloxycoumarin on 5637 cells via induction of apoptosis and cell cycle arrest in G2/M stage, DARU J. Pharm. Sci. 22 (2014) Article ID: 3 (10 pages); DOI: 10.1186/2008-2231-22-3.

24. C. Barthomeuf, S. Lim, M. Iranshahi and P. Chollet, Umbelliprenin from Ferula szowitsiana inhibits the growth of human M4Beu metastatic pigmented malignant melanoma cells through cell-cycle arrest in G1 and induction of caspase-dependent apoptosis, Phytomedicine 15 (2008) 103-111; DOI: 10.1016/j.phymed.2007.04.001.

25. Y. J. Chuang, F. Y. Huang, F. H. Lu, C. H. Ho, S. J. Yang, M. T. Li, W. N. Chang and G. J. Chung, Coumarin induces cell cycle arrest and apoptosis in human cervical cancer HeLa cells through a mitochondria- and caspase-3 dependent mechanism and NF-kappaB down-regulation, In Vivo 21 (2007) 1003-1010.

26. M. Kielbus, K. Skalicka-Wozniak, A. Grabarska, W. Jeleniewicz, M. Dmoszynska-Graniczka, A. Marston, K. Polberg, P. Gawda, J. Klatka and A. Stepulak, 7-Substituted coumarins inhibit proliferation and migration of laryngeal cancer cells in vitro, Anticancer Res. 33 (2013) 4347-4356.

27. D. Yang, T. Gu, T. Wang, Q. Tang and C. Ma, Effects of osthole on migration and invasion in breast cancer cells, Biosci. Biotechnol. Biochem. 74 (2010) 1430-1434; DOI: 1271/bbb.100110.

28. W. F. van der Giesen and L. H. Janssen, Influence of ionization and ion-pair formation on lipophilicity of some 4-hydroxy-coumarin derivatives in the octanol-water system, Int. J. Pharm. 12 (1982) 231-249; DOI: 10.1016/0378-5173(82)90122-3.

29. H. Schönherr and T. Cernak, Profound methyl effects in drug discovery and a call for new C-H methylation reactions, Angew. Chem. Int. Ed. Engl. 52 (2013) 12256-12267; DOI: 10.1002/ anie.201303207.

30. S. C. Leung, S. S. Leung, J. R. Tirado and L. W. Jorgensen, Methyl effects on protein-ligand binding, J. Med. Chem. 55 (2012) 4489-4500; DOI: 10.1021/jm3003697.

Acta Pharmaceutica

The Journal of Croatian Pharmaceutical Society

Journal Information

IMPACT FACTOR 2017: 1.071
5-year IMPACT FACTOR: 1.623

CiteScore 2017: 1.46

SCImago Journal Rank (SJR) 2017: 0.362
Source Normalized Impact per Paper (SNIP) 2017: 0.642

Cited By


All Time Past Year Past 30 Days
Abstract Views 0 0 0
Full Text Views 225 225 25
PDF Downloads 96 96 13