Skip to main content

Advertisement

SpringerLink
Log in

Induction of apoptosis and downregulation of ERα in DMBA-induced mammary gland tumors in Sprague–Dawley rats by synthetic 3,5-disubstituted isoxazole derivatives

  • Published:
Molecular and Cellular Biochemistry Aims and scope Submit manuscript

Abstract

Isoxazole derivatives are an important group of chemotherapeutic prototypes. In the current study, we have synthesized few isoxazole derivatives and tested them for their antiproliferative properties in cancer cell lines such as MCF7 and HeLa. The lead compound, 3-(3,4-dimethoxyphenyl)-5-(thiophen-2-yl)isoxazole (2b), showed considerable inhibition of proliferation of MCF7 and HeLa cells with the IC50 values of 19.5 and 39.2 µM, respectively. Cell cycle analyses and annexin-FITC staining in 2b-treated breast adenocarcinoma cells (MCF7) showed increased sub-G1 population and apoptosis. Furthermore, we tested the tumor inhibitory effect of 2b and estrogen receptor expression profile in DMBA-induced mammary tumors in Sprague–Dawley rats. The gross morphology of tumor studies was investigated by histopathology and ERα protein expression was evaluated by immunohistochemistry, which showed tumor regression and downregulation of ERα in tumor cells. The present results implicate that compound 2b could be used for the further derivatization for the treatment of breast cancer.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

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

Similar content being viewed by others

References

  1. Manson JE, Martin KA (2001) Clinical practice. Postmenopausal hormone-replacement therapy. N Engl J Med 345:34–40

    Article  CAS  PubMed  Google Scholar 

  2. Green S, Walter P, Kumar V, Krust A, Bornert JM, Argos P, Chambon P (1986) Human oestrogen receptor cDNA: sequence, expression and homology to v-erb-A. Nature 320:134–139

    Article  CAS  PubMed  Google Scholar 

  3. Kuiper GG, Enmark E, Pelto-Huikko M, Nilsson S, Gustafsson JA (1996) Cloning of a novel receptor expressed in rat prostate and ovary. Proc Natl Acad Sci USA 93:5925–5930

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  4. Dahlman-Wright K, Cavailles V, Fuqua SA, Jordan VC, Katzenellenbogen JA, Korach KS, Maggi A, Muramatsu M, Parker MG, Gustafsson JA (2006) International union of pharmacology. LXIV. Estrogen receptors. Pharmacol Rev 58:773–781

    Article  CAS  PubMed  Google Scholar 

  5. Moon BS, Carlson KE, Katzenellenbogen JA, Choi TH, Chi DY, Kim JY, Cheon GJ, Koh HY, Lee KC, An G (2009) Synthesis and evaluation of aryl-substituted diarylpropionitriles, selective ligands for estrogen receptor beta, as positron-emission tomographic imaging agents. Bioorg Med Chem 17:3479–3488

    Article  CAS  PubMed  Google Scholar 

  6. Barrett I, Meegan MJ, Hughes RB, Carr M, Knox AJ, Artemenko N, Golfis G, Zisterer DM, Lloyd DG (2008) Synthesis, biological evaluation, structural-activity relationship, and docking study for a series of benzoxepin-derived estrogen receptor modulators. Bioorg Med Chem 16:9554–9573

    Article  CAS  PubMed  Google Scholar 

  7. Yermolina MV, Wang J, Caffrey M, Rong LL, Wardrop DJ (2011) Discovery, synthesis, and biological evaluation of a novel group of selective inhibitors of filoviral entry. J Med Chem 54:765–781

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  8. Vorobyeva DV, Karimova NM, Odinets IL, Roschenthaler GV, Osipov SN (2011) Click-chemistry approach to isoxazole-containing alpha-CF3-substituted alpha-aminocarboxylates and alpha-aminophosphonates. Org Biomol Chem 9:7335–7342

    Article  CAS  PubMed  Google Scholar 

  9. Volkova YA, Averina EB, Grishin YK, Bruheim P, Kuznetsova TS, Zefirov NS (2010) Unexpected heterocyclization of electrophilic alkenes by tetranitromethane in the presence of triethylamine. Synthesis of 3-nitroisoxazoles. J Org Chem 75:3047–3052

    Article  CAS  PubMed  Google Scholar 

  10. Filippakopoulos P, Knapp S (2014) Targeting bromodomains: epigenetic readers of lysine acetylation. Nat Rev Drug Discov 13:337–356

    Article  CAS  PubMed  Google Scholar 

  11. Sharath Kumar KS, Hanumappa A, Vetrivel M, Hegde M, Girish YR, Byregowda TR, Rao S, Raghavan SC, Rangappa KS (2015) Antiproliferative and tumor inhibitory studies of 2,3 disubstituted 4-thiazolidinone derivatives. Bioorg Med Chem Lett 25:3616–3620

    Article  CAS  PubMed  Google Scholar 

  12. Poutiainen PK, Venalainen TA, Perakyla M, Matilainen JM, Vaisanen S, Honkakoski P, Laatikainen R, Pulkkinen JT (2010) Synthesis and biological evaluation of phenolic 4,5-dihydroisoxazoles and 3-hydroxy ketones as estrogen receptor alpha and beta agonists. Bioorg Med Chem 18:3437–3447

    Article  CAS  PubMed  Google Scholar 

  13. Sharath Kumar KS, Hanumappa A, Hegde M, Narasimhamurthy KH, Raghavan SC, Rangappa KS (2014) Synthesis and antiproliferative effect of novel 4-thiazolidinone-, pyridine- and piperazine-based conjugates on human leukemic cells. Eur J Med Chem 81:341–349

    Article  CAS  PubMed  Google Scholar 

  14. Hegde M, Sharath Kumar KS, Thomas E, Ananda H, Raghavan SC, Rangappa KS (2015) A novel benzimidazole derivative binds to the DNA minor groove and induces apoptosis in leukemic cells. RSC Adv 5:93194–93208

    Article  CAS  Google Scholar 

  15. Kavitha CV, Nambiar M, Ananda Kumar CS, Choudhary B, Muniyappa K, Rangappa KS, Raghavan SC (2009) Novel derivatives of spirohydantoin induce growth inhibition followed by apoptosis in leukemia cells. Biochem Pharmacol 77:348–363

    Article  CAS  PubMed  Google Scholar 

  16. LG L (1968) 3rd edn. McGrawHill, New York

  17. Poola I, Yue Q (2007) Estrogen receptor alpha (ER alpha) mRNA copy numbers in immunohistochemically ER alpha-positive-, and negative breast cancer tissues. BMC Cancer 7:56

    Article  PubMed  PubMed Central  Google Scholar 

  18. Hsu KC, Chen YF, Lin SR, Yang JM (2011) iGEMDOCK: a graphical environment of enhancing GEMDOCK using pharmacological interactions and post-screening analysis. BMC Bioinform 12(Suppl 1):S33

    Article  Google Scholar 

  19. Yang JM, Shen TW (2005) A pharmacophore-based evolutionary approach for screening selective estrogen receptor modulators. Proteins 59:205–220

    Article  PubMed  Google Scholar 

  20. Tikoo K, Kumar P, Gupta J (2009) Rosiglitazone synergizes anticancer activity of cisplatin and reduces its nephrotoxicity in 7, 12-dimethyl benz{a}anthracene (DMBA) induced breast cancer rats. BMC Cancer 9:107

    Article  PubMed  PubMed Central  Google Scholar 

  21. Girish YR, Sharath Kumar KS, Manasa HS, Shashikanth S (2014) ZnO: an ecofriendly green nano catalyst for the synthesis of pyrazole derivatives under aqueous media. J Chin Chem Soc 61:1175–1179

    Article  CAS  Google Scholar 

  22. Narasimhamurthy KH, Chandrappa S (2013) Sharath Kumar KS, Swaroop TR RK and Synthetic utility of propylphosphonic anhydride–DMSO media: an efficient one-pot three component synthesis of 2-arylquinolines. Chem Lett 42:1073–1075

    Article  CAS  Google Scholar 

  23. Narasimhamurthy KH, Chandrappa S, Sharath Kumar KS, Harsha KB, Ananda H, Rangappa KS (2014) Easy access for the synthesis of 2-aryl 2,3-dihydroquinazolin-4(1H)-ones using gem-dibromomethylarenes as synthetic aldehyde equivalent. RSC Adv 4:34479–34486

    Article  CAS  Google Scholar 

  24. Hegde M, Mantelingu K, Swarup HA, Pavankumar CS, Qamar I, Raghavan SC, Rangappa KS (2016) Novel PARP inhibitors sensitize human leukemic cells in an endogenous PARP activity dependent manner. RSC Adv 6:6308–6319

    Article  CAS  Google Scholar 

  25. Raghava B, Parameshwarappa G, Acharya A, Swaroop TR, Rangappa KS, Illa H (2014) Cyclocondensation of hydroxylamine with 1,3-bis(het)arylmonothio 1,3-diketones and 1,3-bis(het)aryl-3-(methylthio)-2-propenones: synthesis of 3,5-bis(het)arylisoxazoles with complementary regioselectivity. Eur J Org Chem 9:1882–1892

    Article  Google Scholar 

  26. Baburajeev CP, Dhananjaya Mohan C, Ananda H, Rangappa S, Fuchs JE, Jagadish S, Sivaraman Siveen K, Chinnathambi A, Ali Alharbi S, Zayed ME, Zhang J, Li F, Sethi G, Girish KS, Bender A, Basappa Rangappa KS (2015) Development of novel triazolo-thiadiazoles from heterogeneous “green” catalysis as protein tyrosine phosphatase 1B inhibitors. Sci Rep 5:14195

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  27. Hewings DS, Fedorov O, Filippakopoulos P, Martin S, Picaud S, Tumber A, Wells C, Olcina MM, Freeman K, Gill A, Ritchie AJ, Sheppard DW, Russell AJ, Hammond EM, Knapp S, Brennan PE, Conway SJ (2013) Optimization of 3,5-dimethylisoxazole derivatives as potent bromodomain ligands. J Med Chem 56:3217–3227

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  28. Hewings DS, Wang M, Philpott M, Fedorov O, Uttarkar S, Filippakopoulos P, Picaud S, Vuppusetty C, Marsden B, Knapp S, Conway SJ, Heightman TD (2011) 3,5-dimethylisoxazoles act as acetyl-lysine-mimetic bromodomain ligands. J Med Chem 54:6761–6770

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  29. Li X, Huang J, Yi P, Bambara RA, Hilf R, Muyan M (2004) Single-chain estrogen receptors (ERs) reveal that the ERalpha/beta heterodimer emulates functions of the ERalpha dimer in genomic estrogen signaling pathways. Mol Cell Biol 24:7681–7694

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  30. Sdelci S, Lardeau CH, Tallant C, Klepsch F, Klaiber B, Bennett J, Rathert P, Schuster M, Penz T, Fedorov O, Superti-Furga G, Bock C, Zuber J, Huber KV, Knapp S, Muller S, Kubicek S (2016) Mapping the chemical chromatin reactivation landscape identifies BRD4-TAF1 cross-talk. Nat Chem Biol 12:504–510

    Article  CAS  PubMed  Google Scholar 

  31. Shen DW, Pouliot LM, Hall MD, Gottesman MM (2012) Cisplatin resistance: a cellular self-defense mechanism resulting from multiple epigenetic and genetic changes. Pharmacol Rev 64:706–721

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  32. Montenegro RC, Clark PG, Howarth A, Wan X, Ceroni A, Siejka P, Nunez-Alonso GA, Monteiro O, Rogers C, Gamble V, Burbano R, Brennan PE, Tallant C, Ebner D, Fedorov O, O’Neill E, Knapp S, Dixon D, Muller S (2016) BET inhibition as a new strategy for the treatment of gastric cancer. Oncotarget. doi:10.18632/oncotarget.9766

    PubMed  Google Scholar 

  33. Poma P, Notarbartolo M, Labbozzetta M, Maurici A, Carina V, Alaimo A, Rizzi M, Simoni D, D’Alessandro N (2007) 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 20:329–335

    CAS  PubMed  Google Scholar 

  34. Mohibi S, Mirza S, Band H, Band V (2011) Mouse models of estrogen receptor-positive breast cancer. J Carcinog 10:35

    Article  PubMed  PubMed Central  Google Scholar 

  35. Chatterjee S, Malhotra R, Varghese F, Bukhari AB, Patil A, Budrukkar A, Parmar V, Gupta S, De A (2013) Quantitative immunohistochemical analysis reveals association between sodium iodide symporter and estrogen receptor expression in breast cancer. PLoS One 8:e54055

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  36. Rochefort H, Borgna JL, Evans E (1983) Cellular and molecular mechanism of action of antiestrogens. J Steroid Biochem 19:69–74

    Article  CAS  PubMed  Google Scholar 

  37. Warri AM, Huovinen RL, Laine AM, Martikainen PM, Harkonen PL (1993) Apoptosis in toremifene-induced growth inhibition of human breast cancer cells in vivo and in vitro. J Natl Cancer Inst 85:1412–1418

    Article  CAS  PubMed  Google Scholar 

  38. Somai S, Chaouat M, Jacob D, Perrot JY, Rostene W, Forgez P, Gompel A (2003) Antiestrogens are pro-apoptotic in normal human breast epithelial cells. Int J Cancer 105:607–612

    Article  CAS  PubMed  Google Scholar 

  39. Thiantanawat A, Long BJ, Brodie AM (2003) Signaling pathways of apoptosis activated by aromatase inhibitors and antiestrogens. Cancer Res 63:8037–8050

    CAS  PubMed  Google Scholar 

Download references

Acknowledgments

H. A. thanks ICMR for SRF No. 45/54/2013-PHA/BMS, Dated: 08.12.2014 for research fellowship. We also thank Dr. Suguna Rao, Dr. Nishana M, Dr. Robin Sebastian, and other members of Sahees C. Raghavan group, Department of Biochemistry, Indian Institute of Science, Bangalore for suggestions and critical reading of the manuscript.

Author information

Author notes

    Authors and Affiliations

    1. Department of Studies in Chemistry, Manasagangotri University of Mysore, Mysore, 570006, India

      Hanumappa Ananda, Mahesh Hegde & Kanchugarakoppal S. Rangappa

    2. Department of Chemistry, Regional Institute of Education (NCERT, New Delhi), Manasagangotri, Mysore, 570006, India

      Kothanahally S. Sharath Kumar

    Authors
    1. Hanumappa Ananda
      View author publications

      You can also search for this author in PubMed Google Scholar

    2. Kothanahally S. Sharath Kumar
      View author publications

      You can also search for this author in PubMed Google Scholar

    3. Mahesh Hegde
      View author publications

      You can also search for this author in PubMed Google Scholar

    4. Kanchugarakoppal S. Rangappa
      View author publications

      You can also search for this author in PubMed Google Scholar

    Corresponding author

    Correspondence to Kanchugarakoppal S. Rangappa.

    Additional information

    H. Ananda and K. S. S. Kumar have contributed equally to this work.

    Rights and permissions

    Reprints and permissions

    About this article

    Check for updates. Verify currency and authenticity via CrossMark

    Cite this article

    Ananda, H., Kumar, K.S.S., Hegde, M. et al. Induction of apoptosis and downregulation of ERα in DMBA-induced mammary gland tumors in Sprague–Dawley rats by synthetic 3,5-disubstituted isoxazole derivatives. Mol Cell Biochem 420, 141–150 (2016). https://doi.org/10.1007/s11010-016-2777-z

    Download citation

    • Received:

    • Accepted:

    • Published:

    • Issue Date:

    • DOI: https://doi.org/10.1007/s11010-016-2777-z

    Keywords

    Search

    Navigation