Antiangiogenic and Anticancer Potential of Supercritical Fluid Extracts from Nutmeg Seeds; In vitro, Ex vivo and In silico studies
Sawsan S. Al-Rawi1*, Ahmad Hamdy Ibrahim2, Marthad A. Hamde3, Dinesh Babu4, Mansoureh Nazari5, Mohd Omar Ab Kadir6, Aman Shah Abdul Majid7, Amin Malik Shah8
Journal of Angiotherapy 7(1) 1-13 https://doi.org/10.25163/angiotherapy.719371
Submitted: 25 October 2023 Revised: 12 November 2023 Published: 14 November 2023
Abstract
Background: Angiogenesis is a vital process of forming new blood vessels that occurs during several normal physiological processes. It plays a substantial role in tumor growth and cancer by supplying oxygen and nutrients for the proliferating tumors. Nutmeg, the dried seed of Myristica fragrans, is known for its therapeutic properties. This study aimed to investigate the antiangiogenic and cytotoxic properties of nutmeg extracts derived by Soxhlet and supercritical fluid extraction (SFE) using in-vitro, ex-vivo, and in-silico studies. Method: Nutmeg anticancer property was evaluated against breast cancer cell MCF7 and colon cancer cell HCT116 using MTT in-vitro assay. The antiangiogenic property was investigated using 3D ex-vivo rat aorta assays. The chemical composition of nutmeg extract was characterized using GC/TOF-MS. Subsequently, the main compounds in the nutmeg extract were analyzed against the angiogenesis-associated molecules (COX-1, VEGFA, HIF, and EGF) by molecular docking and were compared with tamoxifen and 5-fluorouracil. Results: The SFE extracts exhibited higher antiangiogenic properties than the Soxhlet (IC50 31µg/mL). Nutmeg SFE extract exhibited higher cytotoxicity towards HCT116 than MCF7 cells. Several active compounds, including myristicin, eugenol, safrole, and α-asarone were identified in the nutmeg SFE extracts using GC/TOF-MS. Molecular docking revealed strong interactions between these compounds and angiogenesis molecular mediators. Particularly, myristicin blocked COX-1, VEGFA, HIF, and EGF enzymes, indicating possible binding interactions. Conclusion: This study highlights the limitless possibilities of nature's offerings in advancing human health. Nutmeg's potential compounds lie in their binding models, which have proven to be powerful tools against cancer and angiogenesis. Myristicin, α-asarone, safrole, and eugenol work synergistically to induce antiangiogenic effects, making nutmeg a promising natural source of angiogenesis inhibitors for future anticancer therapies. The Molecular docking result confirmed that the inhibition of COX-1, VEGF-A, HIF, and EGF by nutmeg compounds offers great potential in the treatment and prevention of various angiogenesis diseases.
Keywords: Angiogenesis; COX-1; VEGF; HIF; EGF; Cytotoxicity; Cancer; MCF7; HCT116; GC-TOF/ MS, and Nutmeg.
References
Ab Rahman NN, Al-Rawi SS, Ibrahim AH, Nama MMB, Ab Kadir MO. Supercritical carbon dioxide extraction of the residual oil from palm kernel cake. J Food Eng. 108(1), 166–70, (2012).
Ab Rahman NN, Nama MM, Al-Rawi SS, Ibrahim AH, Ab Kadir MO. Comparison of nutritional composition between palm kernel fibre and the effect of the supercritical fluid extraction on its quality. Procedia Food Science. 1, 1, 1940-5(2011).
Aggarwal BB, Van Kuiken ME, Iyer LH, Harikumar KB, Sung B. Molecular targets of nutraceuticals derived from dietary spices: potential role in suppression of inflammation and tumorigenesis. Experimental Biology and Medicine. 234(8), 825-49, (2009)
Al-Ostoot FH, Salah S, Khamees HA, Khanum SA. Tumor angiogenesis: Current challenges and therapeutic opportunities. Cancer Treat Res Commun, 28, 100422 (2021).
Al-Rawi SS, Ibrahim AH, Ab Rahman NNN, Nama MMB, Majid AMA, Ab Kadir MO. The effect of supercritical fluid extraction parameters on the nutmeg oil extraction and its cytotoxic and antiangiogenic properties. Procedia Food Sci, 1, 1946–52 (2011).
Al-Rawi SS, Ibrahim AH, Majid ASA, Majid AMA, Ab Kadir MO. Comparison of yields and quality of nutmeg butter obtained by extraction of nutmeg rind by soxhlet and supercritical carbon dioxide (SC-CO2). J Food Eng. 119(3), 595–601, (2013).
Baldo P, Fornasier G, Francescon S, Ferrarin E, Truccolo I, De Paoli P. Toxicities and adverse drug reactions experienced during anticancer treatment: it is desirable to consider the problem within the international system of pharmacovigilance. J Clin Oncol, 33(25), 2824–5 (2015).
Bar FM, Khanfar MA, Elnagar AY, Badria FA, Zaghloul AM, Ahmad KF, Sylvester PW, El Sayed KA. Design and pharmacophore modeling of biaryl methyl eugenol analogs as breast cancer invasion inhibitors. Bioorganic & medicinal chemistry. 15, 18(2), 496-507 (2010).
Barceloux, D. G. Nutmeg (Myristica fragrans Houtt). Disease-a-Month. Foodborne and Microbial Toxins, Part II: Staples and Spices, 55, 6, 373-379 (2009).
Barman R, Bora PK, Saikia J, Kemprai P, Saikia SP, Haldar S, et al. Nutmegs and wild nutmegs: An update on ethnomedicines, phytochemicals, pharmacology, and toxicity of the Myristicaceae species. Phytother Res, 35(9), 4632–59, (2021).
Chen, W. H., Chen, C. H., Chang, C. M.J., Liau, B. C., Hsiang, D. Supercritical carbon dioxide extraction of triglycerides from Aquilaria crassna seeds. Separation and Purification Technology, 73: 135–141, (2010).
Crozier, A. Clifford, M. N., Ashihara, H. Plant Secondary Metabolites Occurrence, Structure and Role in the Human Diet. Blackwell Publishing Ltd, 9600 Garsington Road, Oxford OX4 2DQ, UK. (2006).
Dahham SS, Al-Rawi SS, Ibrahim AH, Majid ASA, Majid AMSA. Antioxidant, anticancer, apoptosis properties and chemical composition of black truffle Terfezia claveryi. Saudi J Biol Sci. 25(8), 1524–34, (2018)
Diniz C, Suliburska J, Ferreira IMPLVO. New insights into the antiangiogenic and proangiogenic properties of dietary polyphenols. Mol Nutr Food Res. 61(6). doi: 10.1002/mnfr.201600912. PMID: 27981783. (2017)
Ha MT, Vu NK, Tran TH, Kim JA, Woo MH, Min BS. Phytochemical and pharmacological properties of Myristica fragrans Houtt.: an updated review. Arch Pharm Res, 43, 1067–92 (2020).
Habib A, Nazari M, Iqbal MA, Bhatti HN, Ahmed MK, Majid AA. Unsymmetrically substituted benzimidazolium based Silver (I)-N-heterocyclic carbene complexes: Synthesis, characterization and in vitro anticancer study against human breast cancer and colon cancer. J Saudi Chem Soc. 23(7), 795–808, (2019)
Hanif, A., Ibrahim, A. H., Ismail, S., Al-Rawi, S. S., Ahmad, J. N., Hameed, M., ... & Tanwir, S. Cytotoxicity against A549 Human Lung Cancer Cell Line via the Mitochondrial Membrane Potential and Nuclear Condensation Effects of Nepeta paulsenii Briq., a Perennial Herb. Molecules, 28(6), 2812, (2023).
Ibrahim A, Al-Rawi S, Abdul Majid A, Al-Habib O, Abdul Majid A. Pro-angiogenic and wound healing potency of virgin coconut oil. Supp Care Cancer, MASCC. 21:235, (2013).
Ibrahim AH, Al-Rawi SS, Majid AA, Rahman NA, Abo-Salah KM, Ab Kadir MO. Separation and fractionation of Aquilaria malaccensis oil using supercritical fluid extraction and the cytotoxic properties of the extracted oil. Procedia Food Science. 1, 1, 1953-9 (2011).
Ibrahim AH, Al-Rawi SS. Evaluating the supercritical extraction effects on the chemical compositions of the extracted oil using GC/TOF MS. In IOP Publishing, 012166, (2018)
Ibrahim AH, Khan MSS, Al-Rawi SS, Ahamed MBK, Majid ASBA, Al-Suede FSR, et al. Safety assessment of widely used fermented virgin coconut oil (Cocos nucifera) in Malaysia: Chronic toxicity studies and SAR analysis of the active components. Regul Toxicol Pharmacol. 81, 457–679, (2016).
Ibrahim, A. H., Li, H., Al-Rawi, S. S., Majid, A. S. A., Al-Habib, O. A., Xia, X. & Ji, D. Angiogenic and wound healing potency of fermented virgin coconut oil: in vitro and in vivo studies. American Journal of Translational Research, 9(11), 4936, (2017).
Jaiswal P, Kumar P, Singh VK, Singh DK. Biological Effects of Myristica fragrans. Annual Review of Biomedical Sciences, 11, 21-29, (2009).
Khan T, Ali M, Khan A, Nisar P, Jan SA, Afridi S, et al. Anticancer plants: A review of the active phytochemicals, applications in animal models, and regulatory aspects. Biomolecules. 10(1), 47, (2019)
Kim, H. Flavonoids inhibit VEGF-bFGF-induced angiogenesis in vitro by inhibiting the matrix-degrading proteases. Journal of Cellular Biochemistry, 89, 529-538 (2003).
King, J.W. Supercritical Fluid Extraction: Present Status and Prospects. Grasas y Aceites. 53. 1, 8-21, (2002).
Lee W, Patel JH, Lockhart AC. Novel targets in esophageal and gastric cancer: beyond antiangiogenesis. Expert opinion on investigational drugs. 1, 18(9), 1351-64 (2009).
Lugano R, Ramachandran M, Dimberg A. Tumor angiogenesis: causes, consequences, challenges and opportunities. Cell Mol Life Sci, 77:1745–70, (2020).
Mattiuzzi C, Lippi G. Current cancer epidemiology. J Epidemiol Glob Health, 9(4), 217, (2019)
Miller, N.J. and Ruiz-Larrea, M.B. Flavonoids and other plant phenols in the diet: Their significance as antioxidants. Journal of Nutritional and Environmental Medicine. 12, 39-47, (2002).
Montemagno C, Pagès G. Resistance to anti-angiogenic therapies: a mechanism depending on the time of exposure to the drugs. Front Cell Dev Biol, 8, 584 (2020).
Morbidelli, L., Terzuoli, E., & Donnini, S. (2018). Use of nutraceuticals in angiogenesis-dependent disorders. Molecules, 23(10), 2676.
Mukund V, Saddala MS, Farran B, Mannavarapu M, Alam A, Nagaraju GP. Molecular docking studies of angiogenesis target protein HIF-1 alpha and genistein in breast cancer. Gene. 15, 70, 169-172, (2019).
Murray, M.T., Pizzorno, J., Pizzorno, L. The Encyclopedia of Healing Foods. Atria Books, New York: 10020. (2005).
Nazari M, Mahmood S, Raman S. Comparative docking studies of rosmarinic acid and sinesitin to inhibit HSP70. International Journal of Engineering Technology and Sciences. 10, 6(1), 115–9, (2019)
Nazari M, Nazari M, Arabani S. Anti-Inflammation Prediction of Orthosiphon Stamineus Extract Against Covid19 (In Silico Study). International Journal of Engineering Technology and Sciences. 23, 8(1), 14-8, (2021)
Rust R. Insights into the dual role of angiogenesis following stroke. J Cereb Blood Flow Metab, 40(6), 1167–71, (2020)
Seneme EF, Dos Santos DC, Silva EM, Franco YE, Longato GB. Pharmacological and therapeutic potential of myristicin: A literature review. Molecules. 29, 26(19), 5914, (2021)
Shanmugam MK, Warrier S, Kumar AP, Sethi G, Arfuso F. Potential Role of Natural Compounds as Anti-Angiogenic Agents in Cancer. Curr Vasc Pharmacol.15(6), 503–19 (2017).
Sim, H. W., Wachsmuth, L., Barnes, E. H., Yip, S., Koh, E. S., Hall, M., ... & Khasraw, M. (2023). NUTMEG: A randomized phase II study of nivolumab and temozolomide versus temozolomide alone in newly diagnosed older patients with glioblastoma. Neuro-Oncology Advances, 5(1), vdad124.
Usui, K., Kubota, E., Kobayashi, H., Fujita, Y., Hatanaka, K., Kamijo, Y., ... & Mimasaka, S. (2023). Detection of major psychoactive compounds (safrole, myristicin, and elemicin) of nutmeg in human serum via GC–MS/MS using MonoSpin® extraction: Application in a nutmeg poisoning case. Journal of Pharmaceutical and Biomedical Analysis, 234, 115565.
Wang H, Oo Khor T, Shu L, Su ZY, Fuentes F, Lee JH, et al. Plants vs. cancer: a review on natural phytochemicals in preventing and treating cancers and their druggability. Anti-Cancer Agents in Medicinal Chemistry (Formerly Current Medicinal Chemistry-Anti-Cancer Agents), 12(10), 1281-1305. (2012).
Xiao JB, Chen JW, Xu M. Xiao JB, Chen JW, Xu M. Supercritical fluid CO2 extraction of essential oil from Marchantia convoluta: global yields and extract chemical composition. Electronic Journal of Biotechnology. 10(1):141–8, (2007).
Yang T, Xiao H, Liu X, Wang Z, Zhang Q, Wei N, et al. Vascular normalization: a new window opened for cancer therapies. Front Oncol, 11, 719836 (2021).
Zhang, W. K., Tao, S. S., Li, T. T., Li, Y. S., Li, X. J., Tang, H. B., ... & Wan, C. J. (2016). Nutmeg oil alleviates chronic inflammatory pain through inhibition of COX-2 expression and substance P release in vivo. Food & nutrition research, 60(1), 30849.
Zu, Y., Yu, H., Liang, L., Fu, Y., Efferth, T., Liu, X., & Wu, N. Activities of ten essential oils towards Propionibacterium acnes and PC-3, A-549 and MCF-7 cancer cells. Molecules. 15(5), 3200–10, (2010).
View Dimensions
View Altmetric
Save
Citation
View
Share