Cytotoxic Activity of a Niclosamide–Caffeic Acid Multicomponent Cocrystal in T47D Breast Cancer Cells
Abstract
Objective: This study aimed to evaluate the cytotoxic activity of a niclosamide–caffeic acid multicomponent cocrystal against T47D breast cancer cells.
Methods: The multicomponent cocrystal was prepared using the solvent-drop grinding method. Cytotoxic activity was assessed using the Microculture Tetrazolium Test (MTT) assay.
Results: The results demonstrated that both niclosamide alone and the niclosamide–caffeic acid multicomponent cocrystal exhibited cytotoxic effects against T47D cells, with IC50 values of 2.92 μg/mL and 2.50 μg/mL, respectively. Microplate spectrophotometric analysis confirmed a concentration-dependent decrease in cell viability for both treatments. Notably, the niclosamide–caffeic acid multicomponent cocrystal displayed a more pronounced cytotoxic effect than niclosamide alone.
Conclusions: These findings suggest that the formation of niclosamide–caffeic acid multicomponent crystals may enhance anticancer potential and warrant further investigation as a promising therapeutic approach for breast cancer treatment.
Keywords: breast cancer, niclosamide, caffeic acid, multicomponent crystal, cytotoxicity, T47D cells, MTT assay.
Keywords:
breast cancer, niclosamide, caffeic acid, multicomponent crystal, cytotoxicity, T47D cells, MTT assayDOI
https://doi.org/10.22270/jddt.v15i9.7363References
1. Sung H, Ferlay J, Siegel RL, Laversanne M, Soerjomataram I, Jemal A, et al. Global cancer statistics 2020: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 Countries. CA Cancer J Clin. 2021;71(3):209-49. https://doi.org/10.3322/caac.21660 PMid:33538338
2. Kim J, Harper A, McCormack V, Sung H, Houssami N, Morgan E, et al. Global patterns and trends in breast cancer incidence and mortality across 185 countries. Nat Med. 2025;31(4):1154-62. https://doi.org/10.1038/s41591-025-03502-3 PMid:39994475
3. Perkins DW, Steiner I, Haider S, Robertson D, Buus R, O'Leary L, et al. Therapy-induced normal tissue damage promotes breast cancer metastasis. iScience. 2024;27(1):1-21. https://doi.org/10.1016/j.isci.2023.108503 PMid:38161426 PMCid:PMC10755366
4. Navya PN, Kaphle A, Srinivas SP, Bhargava SK, Rotello VM, Daima HK. Current trends and challenges in cancer management and therapy using designer nanomaterials. Nano Converg. 2019;6(1):1-30. https://doi.org/10.1186/s40580-019-0193-2 PMid:31304563 PMCid:PMC6626766
5. Saranraj K, Kiran PU. Drug repurposing: Clinical practices and regulatory pathways. Perspect Clin Res. 2025;16(2):61-8. https://doi.org/10.4103/picr.picr_70_24 PMid:40322475 PMCid:PMC12048090
6. Xia Y, Sun M, Huang H, Jin WL. Drug repurposing for cancer therapy. Signal Transduct Target Ther. 2024;9(1):92. https://doi.org/10.1038/s41392-024-01808-1 PMid:38637540 PMCid:PMC11026526
7. Yin L, Gao Y, Zhang X, Wang J, Ding D, Zhang Y, et al. Niclosamide sensitizes triple-negative breast cancer cells to ionizing radiation in association with the inhibition of Wnt/β-catenin signaling. Oncotarget. 2016;7(27):42126-38. https://doi.org/10.18632/oncotarget.9704 PMid:27363012 PMCid:PMC5173121
8. Wu MM, Zhang Z, Tong CWS, Yan VW, Cho WCS, To KKW. Repurposing of niclosamide as a STAT3 inhibitor to enhance the anticancer effect of chemotherapeutic drugs in treating colorectal cancer. Life Sci. 2020;262:118522. https://doi.org/10.1016/j.lfs.2020.118522 PMid:33011217
9. Laila UE, Zhao Z long, Xu DY, Liu H, Xu ZX. Pharmacological advances and therapeutic applications of niclosamide in cancer and other diseases. Eur J Med Chem. 2025;290:117527. https://doi.org/10.1016/j.ejmech.2025.117527 PMid:40153934
10. Pan JX, Ding K, Wang CY. Niclosamide, an old antihelminthic agent, demonstrates antitumor activity by blocking multiple signaling pathways of cancer stem cells. Chin J Cancer. 2012;31(4):178-84. https://doi.org/10.5732/cjc.011.10290 PMid:22237038 PMCid:PMC3777479
11. Wiggins R, Woo J, Mito S. Optimizing Niclosamide for Cancer Therapy: Improving Bioavailability via Structural Modification and Nanotechnology. Cancers (Basel). 2024;16(20):3548. https://doi.org/10.3390/cancers16203548 PMid:39456642 PMCid:PMC11506536
12. Li Y, Li PK, Roberts MJ, Arend RC, Samant RS, Buchsbaum DJ. Multi-targeted therapy of cancer by niclosamide: A new application for an old drug. Cancer Lett. 2014;349(1):8-14. https://doi.org/10.1016/j.canlet.2014.04.003 PMid:24732808 PMCid:PMC4166407
13. Sathisaran I, Dalvi SV. Engineering Cocrystals of PoorlyWater-Soluble Drugs to Enhance Dissolution in Aqueous Medium. Pharmaceutics. 2018 Jul;10(3). https://doi.org/10.3390/pharmaceutics10030108 PMid:30065221 PMCid:PMC6161265
14. Chettri A, Subba A, Singh GP, Bag PP. Pharmaceutical co-crystals: A green way to enhance drug stability and solubility for improved therapeutic efficacy. J Pharm Pharmacol. 2024;76(1):1-12. https://doi.org/10.1093/jpp/rgad097 PMid:37934904
15. Pantwalawalkar J, Kale N, Nangare S, Patil S, Pawar S, Jadhav N. Pharmaceutical cocrystals: Unlocking the potential of challenging drug candidates. J Drug Deliv Sci Technol. 2025;104:106572. https://doi.org/10.1016/j.jddst.2024.106572
16. Fitriani L, Dirfedli F, Yuliandra Y, Setyawan D, Uchida M, Oyama H, et al. A novel cocrystal approach celecoxib with piperine: Simultaneously enhance dissolution rate and compressibility. J Pharm Sci. 2024;113(12):3565-73. https://doi.org/10.1016/j.xphs.2024.10.011 PMid:39414077
17. Zaini E, Sumirtapura YC, Halim A, Fitriani L, Soewandhi SN. Formation and characterization of sulfamethoxazole-trimethoprim cocrystal by milling process. J Appl Pharm Sci. 2017;7(12):169-73.
18. Kuminek G, Cao F, Bahia de Oliveira da Rocha A, Gonçalves Cardoso S, Rodríguez-Hornedo N. Cocrystals to facilitate delivery of poorly soluble compounds beyond-rule-of-5. Adv Drug Deliv Rev. 2016;101:143-66. https://doi.org/10.1016/j.addr.2016.04.022 PMid:27137109 PMCid:PMC4910885
19. Liang X, Liu S, Li Z, Deng Y, Jiang Y, Yang H. Efficient cocrystal coformer screening based on a Machine learning Strategy: A case study for the preparation of imatinib cocrystal with enhanced physicochemical properties. Eur J Pharm Biopharm [Internet]. 2024;196:114201. Available from: https://www.sciencedirect.com/science/article/pii/S0939641124000274 https://doi.org/10.1016/j.ejpb.2024.114201 PMid:38309538
20. Barbosa S, Agostini N, Borges A, Araújo B De, Bonfilio R. Growing Interest in Pharmaceutical Cocrystals : A Comprehensive Review of Applications and Trends. Chem Sel. 2025;1(00831):1-43. https://doi.org/10.1002/slct.202500831
21. Ifora I, Hamidi D, Susanti M, Hefni D, Wahyuni FS. Enhancing Chemotherapeutic Efficacy: Synergistic Cytotoxic Effect of Garcinia cowa Bark Extract and Doxorubicin in T47D Breast Cancer Cells. Trop J Nat Prod Res. 2025 Jan 31;9(1 SE-Articles):67-72. https://doi.org/10.26538/tjnpr/v9i1.10
22. Sanphui P, Kumar SS, Nangia A. Pharmaceutical Cocrystals of Niclosamide. Cryst Growth Des. 2012;12(9):4588-99. https://doi.org/10.1021/cg300784v
23. Grifasi F, Chierotti MR, Gaglioti K, Gobetto R, Maini L, Braga D, et al. Using salt cocrystals to improve the solubility of niclosamide. Cryst Growth Des. 2015;15(4):1939-48. https://doi.org/10.1021/acs.cgd.5b00106
24. Emami S, Siahi-Shadbad M, Adibkia K, Barzegar-Jalali M. Recent advances in improving oral drug bioavailability by cocrystals. Bioimpacts. 2018;8(4):305-20. https://doi.org/10.15171/bi.2018.33 PMid:30397585 PMCid:PMC6209825
25. Sandra F, Sidharta MA. Caffeic Acid Induced Apoptosis in MG63 Osteosarcoma Cells Through Activation of Caspases. Mol Cell Biomed Sci. 2017;1(1):28. https://doi.org/10.21705/mcbs.v1i1.6
26. Cortez N, Villegas C, Burgos V, Cabrera-Pardo JR, Ortiz L, González-Chavarría I, et al. Adjuvant Properties of Caffeic Acid in Cancer Treatment. Vol. 25, International Journal of Molecular Sciences. 2024. https://doi.org/10.3390/ijms25147631 PMid:39062873 PMCid:PMC11276737
27. Alam M, Ashraf GM, Sheikh K, Khan A, Ali S, Ansari MM, et al. Potential Therapeutic Implications of Caffeic Acid in Cancer Signaling: Past, Present, and Future. Front Pharmacol. 2022;13(March):1-14. https://doi.org/10.3389/fphar.2022.845871 PMid:35355732 PMCid:PMC8959753
28. Balc-Okcanoğlu T, Yilma-Susluer S, Kayabasi C, Ozme-Yelken B, Biray-Avci C, Gunduz C. The effect of caffeic acid phenethyl ester on cell cycle control gene expressions in breast cancer cells. Mol Biol Res Commun. 2021 Mar;10(1):39-43.
29. Chen Y, Li X, Yang M, Liu SB. Research progress on morphology and mechanism of programmed cell death. Cell Death Dis. 2024;15(5):327. https://doi.org/10.1038/s41419-024-06712-8 PMid:38729953 PMCid:PMC11087523
30. Doonan F, Cotter TG. Morphological assessment of apoptosis. Methods. 2008;44(3):200-4. https://doi.org/10.1016/j.ymeth.2007.11.006 PMid:18314050
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