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Isolation and Structural Characterization of Bioactive Compounds from Nigerian Hibiscus Sabdariffa Leaves with Potential Breast Cancer Activity

Jemimah S. Wakawa, Sarah A. Nwinee, Olutayo O. Olajide, Bassi Jirama Ali, Bilkis A., A. Adedoyin

Abstract

Cancer is a disease that occurs when some body cells proliferate out of control and spread to other parts of the body. Current treatments, such as surgery, radiation, chemotherapy, and hormone therapy, have side effects, and might cause multidrug resistance which complicates their effectiveness. This study investigates the cytotoxic potential of crude extracts from Hibiscus sabdariffa leaves and isolates bioactive compounds with relevance to triple-negative breast cancer therapy. The cytotoxicity assays using Brine-Shrine Lethality test and MTT test were carried out on the crude extracts. The BSLT result showed cytotoxicity across all extracts with LC50 values (μg/L) for the n-hexane, ethyl acetate, and methanol extracts were determined as 72.26 (83.79-491.63), 51.19 (42.13-558.11), and 50.61 (43.18-458.12), respectively. The MTT tests, demonstrated significant toxicity across all extracts, notably the methanol and ethyl acetate extracts against several breast cancer cell lines (MCF7, HS578T, SKBr3, MDA-MB-231). Among these, the methanol extract exhibited strong cytotoxicity, with IC50 values 66 μ/M outperforming that of the reference drug Etoposide 78 μ/M against HS578T cell line. Bioassay-guided fractionation and spectroscopic analysis (FTIR and NMR) led to the identification of two compounds: 6-Methoxyquinoline (HS1 or J1) and Ethyl 4-Hydroxybenzoate (HS2 or J2). The thin layer chromatography was carried out to investigate the purity of the different isolates obtained from column chromatography. The compounds where isolated as pure and coded as (HS1 or J1) and (HS2 or J2) which gave a single spot on the chromatoplate. These compounds demonstrated structural and functional characteristics consistent with anticancer, antioxidant, and antimicrobial activities. 6-Methoxyquinoline, a nitrogen-containing heterocyclic compound, exhibited features associated with apoptosis induction and cell cycle inhibition, while Ethyl 4- Hydroxybenzoate demonstrated phenolic-based free radical scavenging activity. These findings lay the groundwork for developing new plant-base therapeutic agents, such as Hibiscus Sabdariffa, aimed at treating resistant form of breast cancer: triple-negative breast cancer Key Words: Cytotoxicity, Bioactive Compounds, Hibiscus Sabdariffa, TNBC Breast Cancer. ,

Keywords

CytotoxicityBioactive CompoundsHibiscus SabdariffaTNBC Breast Cancer.

References

Adinew, G., Messeha, S., & Soliman, K. (2022). Thymoquinone Anticancer Effects Through the Upregulation of NRF2 and the Downregulation of PD-L1 in MDA-MB-231 Triple- Negative Breast Cancer Cells. FASEB Journal: Official Publication of the Federation of American Societies for Experimental Biology. 36. S1.R2586 Ali Esmail Al-Snafi. (2018). Pharmacological and therapeutic importance of Hibiscus Sabdariffa. ResearchGate. 10(3):451-475. Bhattacharya, S., Ghosh, A., & Adhikary, A. (2020). Delivery of thymoquinone through hyaluronic acid-decorated mixed Pluronic® nanoparticles to attenuate angiogenesis and metastasis of triple-negative breast cancer. Journal of Controlled Release: Official Journal of the Controlled Release Society. doi: 10.1016/j.jconrel.2020.03.033 Caccuri, F., Sommariva, M., & Caruso, A. (2019). Inhibition of DNA Repair Mechanisms and Induction of Apoptosis in Triple Negative Breast Cancer Cells Expressing the Human Herpesvirus 6 U94. Cancers, 11. doi: 10.3390/cancers11071006. Daud, S. M., Yaacob, N., & Fauzi, A. (2020). 2-Methoxy-1,4-Naphthoquinone Inhibits Glucose Uptake and Lactate Production in Triple-Negative Breast Cancer Cells. Asian Pacific Journal of Cancer Prevention: APJCP, 22 S1, 59–65. doi: 10.22034/APJCP.2020.21. S2.59 Esra K. A., Haroon K., and Yaseen H. (2022). Herbal Ingredients in the Prevention of Breast Cancer: Comprehensive Review of Potential Molecular Targets and Role of Natural Products. ACS American Chemical Society. Formagio AS., Foglio M.A and Carvalho J.E. (2015). Phenolic compounds of Hibiscus sabdariffa and influence of organic residues on its antioxidant and antitumor properties. PubMed. 75(1):6976. Ganesh N S., Rahul D., and K K Sharma. (2010). Various Types and Management of Breast Cancer: An Overview. Journal of Advance Pharmaceutical Technology and Research. 1(2):109–126. Going, C., Tailor, D., Kumar & Pitteri, S. (2018). Quantitative Proteomic Profiling Reveals Key Pathways in the Anticancer Action of Methoxychalcone Derivatives in Triple Negative Breast Cancer. Journal of Proteome Research, 17 10, 3574–3585. Han, M., Atalay, P., & Küçükgüzel, Ş. (2020). Synthesis, characterization and anticancer activity of novel hydrazide-hydrazones derived from ethyl paraben. Journal of Research in Pharmacy. doi: 10.35333/jrp.2020.156 Hu, M.-H., & Lin, J.-H. (2021). New Dibenzoquinoxalines Inhibit Triple-Negative Breast Cancer Growth by Dual Targeting of Topoisomerase 1 and the c-MYC G-Quadruplex. Journal of Medicinal Chemistry. doi: 10.1021/acs.jmedchem.0c02202 Inês D. R., Ivo P., and Michael H. (2014). Hibiscus sabdariffa L. – A phytochemical and pharmacological review. ELSEVIER. 165, 424-443 Ityo, S., Anhwange, B., Okoye, P., & Feka, P. (2023). Sensory, Gc-Ms And Ftir Analysis of Aqueous Extract of Hibiscus Sabdarriffa And Vernonia Amygdalina Herbal Tea with Blends of Ginger and Lemon Zest. Journal of Chemical Society of Nigeria. doi: 10.46602/jcsn. v48i4.907. Jabeur, I., Pereira, E., & Ferreira, I. (2019). Exploring the chemical and bioactive properties of Hibiscus sabdariffa L. calyces from Guinea-Bissau (West Africa). Food & Function, 10 4, 2234–2243. , Jansi, R., Khusro, A., Agastian, P., Almutairi, M., & Almutairi, B. (2024). Bioassay-guided fractionation and biological activities of antimycin A and 4-hydroxybenzoic acid isolated from Nocardiopsis sp. strain LC-9. BioResources. doi: 10.15376/biores.19.4.7673-7697. Jeannett A. I., Madrigal-B., and Eduardo M.S. (2020) Organic Acids from Roselle (Hibiscus sabdariffa L.) A Brief Review of Its Pharmacological Effects. Biomedicines. 8(5). Kim, H., Kim, S., Sim, G., & Ahn, J.-H. (2020). Synthesis of 4-hydroxybenzoic acid derivatives in Escherichia coli. Journal of Agricultural and Food Chemistry. doi: 10.1021/acs.jafc.0c03149. Lu, W.-J., Huang, Y.-J., & Lin, H. (2022). Phenolic Compound Ethyl 3,4-Dihydroxybenzoate Retards Drug Efflux and Potentiates Antibiotic Activity. Antibiotics, 11. doi: 10.3390/antibiotics11040497. Maciel, L. G., Carmo, M. Do, Azevedo, & Rosso, N. (2018). Hibiscus sabdariffa anthocyanins- rich extract: Chemical stability, in vitro antioxidant and antiproliferative activities. Food and Chemical Toxicology: An International Journal Published for the British Industrial Biological Research Association, 113, 187–197. doi: 10.1016/j.fct.2018.01.053 Mahnashi, M., Alqahtani, Y., & Ayaz, M. (2021). Cytotoxicity, anti-angiogenic, anti-tumor and molecular docking studies on phytochemicals isolated from Polygonum hydropiper L. BMC Complementary Medicine and Therapies, 21. doi: 10.1186/s12906-021-03411-1 National Cancer institute, what is cancer? (2021). https://www.cancer.gov/about- cancer/understanding/what-is-ca Patsorn W., Arunporn I., and Srisopa R. (2014). In vitro antioxidant, anti-inflammatory, cytotoxic activities against prostate cancer of extracts from Hibiscus sabdariffa leaves. PubMed. 8: S81-7. Piyush, M., Rishi, S., Sathiyanarayanan, L., and K. R. Mahadik. (2015). Pharmacokinetic profile of phytoconstituent(s) isolated from medicinal plants-A comprehensive review. Journal of Traditional and Complementary Medicine. 5 Przemystaw J., Boratynski M., and Blajetjacek S. (2019). The Alkaloids: chemistry and Pharmacology. ScienceDirect. pp.1-200.2025 Qotrunnada F., Nadzila A., and Norma N. A. (2020) In vitro Cytotoxicity of Hibiscus sabdariffa Linn Extracts on A549 Lung Cancer Cell Line. Pharmacognosy. 12(1): 1618-1623 Sannino, F., Sansone, C., & Tutino, M. (2018). Pseudoalteromonas haloplanktis TAC125 produces 4-hydroxybenzoic acid that induces pyroptosis in human A459 lung adenocarcinoma cells. Scientific Reports, 8. doi: 10.1038/s41598-018-19536-2. Santosh M., Kurbetti, Ansar M. and Dr Patil. S.S. (2015) Preparation and formulation of polyherbal ointment for wound healing activity. World Journal of Pharmaceutical Research. 4. Shahnaz K., Ahmad S., and Parvin P. (2011). Selective Cytotoxicity and Apoptogenic Activity of Hibiscus Sabdariffa Aqueous Extract Against MCF-7 Human Breast Cancer Cell Line. Journal of Cancer Therapy. 2, 394-400. Susanti, E. (2019). In silico analysis of bioactive compounds of Hibiscus sabdariffa as potential agonists of LXR to inhibit the atherogenesis process. International Conference on Bioinformatics and Nano-Medicine from Natural Resources for Biomedical Research: 3rd Annual Scientific Meeting for Biomedical Sciences. doi: 10.1063/1.5109983. Taib, S. H. M., Shameli, K., & Izadiyan, Z. (2019). Electrooxidation of nitrite based on green synthesis of gold nanoparticles using Hibiscus sabdariffa leaves. Journal of the Taiwan Institute of Chemical Engineers. doi: 10.1016/J.JTICE.2018.09.021. , Ulhaka, K., Kanokwiroon, K., Khongkow, M., Bissanum, R., Khunpitak, T., & Khongkow, P. (2021). The Anticancer Effects of FDI-6, a FOXM1 Inhibitor, on Triple Negative Breast Cancer. International Journal of Molecular Sciences, 22. doi: 10.3390/ijms22136685 Vijayakumar S., Hyunsuk S., and Manikandan R. (2021). Plant Derived Bioactive Compounds, Their Anti-Cancer Effects and in Silicon Approaches as an Alternative Target Treatment Strategy for Breast Cancer. Cancers . 13(24): 6222-6231. World Health Organisation , Breast Cancer. https://www.who.int. 2024. Yuliastri W. O., Ajeng D., and Isrul .M. (2022). Phytochemical Constituent and In-Vitro Cytotoxic Activity of Hibiscus Sabdariffa L. Calyx Fraction on Human Breast Cancer Cell Line MDA-MB-231. ResearchGate: Journal of Chemistry. 15(03):1619-1625

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