Antiviral Effects of Some Flavonoids on SARS-CoV-2

Abstract

Covid-19 disease caused by Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) is a global epidemic that affects millions of lives. To date, there is no definitive cure for the disease and global vaccination efforts with newly produced vaccines will take years to complete. In silico studies have suggested that different flavonoids play an antiviral role against SARS-CoV-2. In this study, based on in silico findings, we examined the in vitro effects of four promising flavonoids, Hesperidin, Oleuropein, Epigallocatechin gallate (EGCG), and Myricetin. First, we extracted these flavonoids from natural plant sources and purified by using liquid chromatography (LC). Next, we analysed the cell toxicity and antiviral activity of these four flavonoids at different concentrations against SARS-CoV-2. Our results using the SARS-CoV-2 infected Vero E6 cell model were found to contradict previous in silico findings, and these flavonoids were found to have no antiviral effects in vitro. Studies investigating the antiviral activity of flavonoids on SARS-CoV-2 should be directed to those other than oleuropein, hesperidin, EGCG, and Myricetin.

Keywords

• Covid-19
• EGCG
• Hesperidin
• Myricetin
• Oleuropein

Project Number

BAP00017

References

1. Adem, S., Eyupoglu, V., Sarfraz, I. et al. (2020). Identification of Potent COVID-19 Main Protease (Mpro) Inhibitors from Natural Polyphenols: An in silico Strategy Unveils a Hope against CORONA. Preprints. doi:10.20944/preprints202003.0333.v1
2. Aouidi, F., Dupuy, N., Artaud, J. et al. (2012). Rapid quantitative determination of oleuropein in olive leaves (Olea europaea) using mid-infrared spectroscopy combined with chemometric analyses. Industrial Crops and Products, 37, 292-297. https://doi.org/10.1016/j.indcrop.2011.12.024
3. Bastolla, U. (2021). Mathematical model of SARS-Cov-2 propagation versus ACE2 fits COVID-19 lethality across age and sex and predicts that of SARS. Frontiers in Molecular Biosciences, 8, 706122. https://doi.org/10.3389/fmolb.2021.706122
4. Boonpawa, R., Spenkelink, A., Punt, A. et al. (2017). Physiologically based kinetic modeling of hesperidin metabolism and its use to predict in vivo effective doses in humans. Molecular Nutrition Food Research, 61(8), 1600894. https://doi.org/10.1002/mnfr.201600894
5. Carneiro, B. M., Batista, M. N., Braga, A. C. S. et al. (2016). The green tea molecule EGCG inhibits Zika virus entry. Virology, 496, 215-218. https://doi.org/10.1016/j.virol.2016.06.012
6. Chacko, S. M., Thambi, P.T., Kuttan, R. et al. (2010). Beneficial effects of green tea: a literature review. Chinese Medicine, 5, 13. https://doi.org/10.1186/1749-8546-5-13
7. Chen, Y. W., Yiu, C-P. B., Wong, K-Y. (2020). Prediction of the SARS-CoV-2 (2019-nCoV) 3C-like protease (3CL pro) structure: virtual screening reveals velpatasvir, ledipasvir, and other drug repurposing candidates. F1000Research, 9, 129. https://doi.org/10.12688/f1000research.22457.2
8. Ciesek, S., Hahn von T., Colpitts, C. et al. (2011). The green tea polyphenol, epigallocatechin-3-gallate, inhibits hepatitis C virus entry. Hepatology, 54(6), 1947-55. https://doi.org/10.1002/hep.24610

9. Cifá, D., Skrt, M., Pittia, P. et al. (2018). Enhanced yield of oleuropein from olive leaves using ultrasound-assisted extraction. Food Science & Nutrition, 6, 1128-37. https://doi.org/10.1002/fsn3.654
10. Clercq, E. De. (2000). Current lead natural products for the chemotherapy of human immunodeficiency virus (HIV) infection. Medicinal Research Reviews, 20, 323-349. https://doi.org/10.1002/1098-1128(200009)20:5<323::AID-MED1>3.0.CO;2-A
11. Conti, C., Genovese, D., Santoro, R. et al. (1990). Activities and mechanisms of action of halogen-substituted flavanoids against poliovirus type 2 infection in vitro. Antimicrobial Agents and Chemotherapy, 34(3), 460-466. https://doi.org/10.1128/AAC.34.3.460
12. Coronaviridae Study Group of the International Committee on Taxonomy of Viruses. (2020). The species severe acute respiratory syndrome-related coronavirus: classifying 2019-nCoV and naming it SARS-CoV-2. Nature Microbiology, 5, 536-544. https://doi.org/10.1038/s41564-020-0695-z
13. Dalluge, J. J., Nelson, B. C., Thomas, J. B. et al. (1998). Selection of column and gradient elution system for the separation of catechins in green tea using high-performance liquid chromatography. Journal of Chromatography A, 793(2), 265-74. https://doi.org/10.1016/S0021-9673(97)00906-0
14. Fang, L., Karakiulakis, G., Roth, M. (2020). Are patients with hypertension and diabetes mellitus at increased risk for COVID-19 infection? The Lancet Respiratory Medicine, 8(4), e21. https://doi.org/10.1016/S2213-2600(20)30116-8
15. Fecka, I., and Turek, S. (2007). Determination of water-soluble polyphenolic compounds in commercial herbal teas from Lamiaceae: peppermint, melissa, and sage. Journal of Agricultural and Food Chemistry, 55(26), 10908-17. https://doi.org/10.1021/jf072284d
16. Genovese, D., Conti, C., Tomao, P. et al. (1995). Effect of chloro-, cyano-, and amidino-substituted flavanoids on enterovirus infection in vitro. Antiviral Research, 27, 123-136. https://doi.org/10.1016/0166-3542(95)00088-4
17. González, Y., Torres-Mendoza, D., Jones, G. E. et al. (2015). Marine Diterpenoids as Potential Anti-Inflammatory Agents. Mediators of Inflammation, 2015, 263543-14. https://doi.org/10.1155/2015/263543
18. Guan, W-J., Ni, Z-Y., Hu, Y. et al. (2020). Clinical Characteristics of Coronavirus Disease 2019 in China. The New England Journal of Medicine, 58(4), 711-712. https://doi.org/10.1016/j.jemermed.2020.04.004
19. Isaacs, C. E., Wen, G. Y., Xu, W. et al. (2008). Epigallocatechin gallate inactivates clinical isolates of herpes simplex virus. Antimicrobial Agents and Chemotherapy, 52, 962-70. https://doi.org/10.1128/AAC.00825-07
20. Iyer, M., Jayaramayya, K., Subramaniam, M. D. et al. (2020). COVID-19: an update on diagnostic and therapeutic approaches. BMB Reports, 53(4), 191-205. https://doi.org/10.5483/BMBRep.2020.53.4.080
21. Khaerunnisa, S., Kurniawan, H., Awaluddin, R. et al. (2020). Potential Inhibitor of COVID-19 Main Protease (Mpro) From Several Medicinal Plant Compounds by Molecular Docking Study. Preprints, 2020030226. doi:10.20944/preprints202003.0226.v1
22. Lahmer, N., Belboukhari, N., Cheriti, K. et al. (2015). Hesperidin and Hesperitin preparation and purification from citrus sinensis peels. Der Pharma Chemica, 7, 1-4.
23. Lee, K. J., and Lee, S. H. (2008). Extraction behavior of caffeine and EGCG from green and black tea. Biotechnology and Bioprocess Engineering, 13, 646-9. https://doi.org/10.1007/s12257-008-0034-3
24. Li, J., Song, D., Wang, S. et al. (2020). Antiviral Effect of Epigallocatechin Gallate via Impairing Porcine Circovirus Type 2 Attachment to Host Cell Receptor. Viruses, 12(2), 176. https://doi.org/10.3390/v12020176
25. Liu, P., Lindstedt, A., Markkinen, N. et al. (2014). Characterization of metabolite profiles of leaves of bilberry (Vaccinium myrtillus L.) and lingonberry (Vaccinium vitis-idaea L.). Journal of Agricultural and Food Chemistry, 62, 12015-26. https://doi.org/10.1021/jf503521m
26. Lu, H. (2020). Drug treatment options for the 2019-new coronavirus (2019-nCoV). BioScience Trends, 14(1), 69-71. https://doi.org/10.5582/bst.2020.01020
27. Lu, R., Zhao, X., Li, J. et al. (2020). Genomic characterisation and epidemiology of 2019 novel coronavirus: implications for virus origins and receptor binding. The Lancet, 395, 565-574. https://doi.org/10.1016/S0140-6736(20)30251-8
28. Mendes, R. A., Almeida, S. K. C., Soares, I. N. et al. (2019). Evaluation of the antioxidant potential of myricetin 3-O-α-L-rhamnopyranoside and myricetin 4'-O-α-L-rhamnopyranoside through a computational study. Journal of Molecular Modeling, 25, 89. https://doi.org/10.1007/s00894-019-3959-x
29. Meneguzzo, F., Ciriminna, R., Zabini, F. et al. (2020). Hydrodynamic Cavitation-based Rapid Expansion of Hesperidin-rich Products from Waste Citrus Peel as a Potential Tool Against COVID-19. Processes 2020, 8, 549. doi:10.20944/preprints202004.0152.v1
30. Narotzki, B., Reznick, A. Z., Aizenbud, D. et al. (2012). Green tea: a promising natural product in oral health. Archives of Oral Biology, 57(5), 429-35. https://doi.org/10.1016/j.archoralbio.2011.11.017
31. Nguyen, T. T. H., Woo, H-J., Kang, H-K. et al. (2012). Flavonoid-mediated inhibition of SARS coronavirus 3C-like protease expressed in Pichia pastoris. Biotechnology Letters, 34, 831-8. https://doi.org/10.1007/s10529-011-0845-8
32. Omar, S. H. (2010). Oleuropein in olive and its pharmacological effects. Scientia Pharmaceutica, 78, 133-54. https://doi.org/10.3797/scipharm.0912-18
33. Peterson, L. (2020). In silico Molecular Dynamics Docking of Drugs to the Inhibitory Active Site of SARS-CoV-2 Protease and Their Predicted Toxicology and ADME. SSRN Journal, http://dx.doi.org/10.2139/ssrn.3580951
34. Rigacci, S., Stefani, M. (2016). Nutraceutical Properties of Olive Oil Polyphenols. An Itinerary from Cultured Cells through Animal Models to Humans. International Journal of Molecular Science, 17(6), 843. https://doi.org/10.3390/ijms17060843
35. Rodriguez-Morales, A. J., Bonilla-Aldana, D. K., Tiwari, R. et al. (2020). COVID-19, an Emerging Coronavirus Infection: Current Scenario and Recent Developments - An Overview. Journal of Pure and Applied Microbiology, 14, 05-12. https://doi.org/10.22207/JPAM.14.1.02
36. Ryu, Y. B., Jeong, H. J., Kim, J. H. et al. (2010). Biflavonoids from Torreya nucifera displaying SARS-CoV 3CLpro inhibition. Bioorganic & Medicinal Chemistry, 18(22), 7940-7. https://doi.org/10.1016/j.bmc.2010.09.035
37. Sahin, S., Demir, C., Malyer, H. (2011). Determination of phenolic compounds in Prunella L. by liquid chromatography-diode array detection. Journal of Pharmaceutical and Biomedical Analysis, 55(5), 1227-30. https://doi.org/10.1016/j.jpba.2011.03.016
38. Silva, S., Gomes, L., Leitão, F. et al. (2016). Phenolic Compounds and Antioxidant Activity of Olea europaea L. Fruits and Leaves. Food Science and Technology International, 12(5), 385-95. https://doi.org/10.1177/1082013206070166
39. Superti, F., Seganti, L., Orsi, N. et al. (1990). In vitro effect of synthetic flavanoids on astrovirus infection. Antiviral Research, 13, 201-208. https://doi.org/10.1016/0166-3542(90)90038-9
40. Umashankar, N., Pemmanda, B., Gopkumar, P. et al. (2018). Effectiveness of topical green tea against multidrug-resistant Staphylococcus aureus in cases of primary pyoderma: An open controlled trial. Indian Journal of Dermatology, Venereology and Leprology, 84, 163-8. doi:10.4103/ijdvl.IJDVL_207_16
41. Wang, S-J., Tong, Y., Lu, S. et al. (2010). Anti-inflammatory activity of myricetin isolated from Myrica rubra Sieb. et Zucc. leaves. Planta Medica, 76(14), 1492-6. doi:10.1055/s-0030-1249780
42. Wei, F., Ma, S-C., Ma, L-Y. et al. (2004). Antiviral flavonoids from the seeds of Aesculus chinensis. Journal of Natural Products, 67(4), 650-653. https://doi.org/10.1021/np030470h
43. Xu, H. X., Lee, S. F. (2001). Activity of plant flavonoids against antibiotic-resistant bacteria. Phytotherapy Research 15, 39-43. https://doi.org/10.1002/1099-1573(200102)15:1<39::AID-PTR684>3.0.CO;2-R
44. Yamaguchi, K., Honda, M., Ikigai, H. et al. (2002). Inhibitory effects of (-)-epigallocatechin gallate on the life cycle of human immunodeficiency virus type 1 (HIV-1). Antiviral Research, 53, 19-34. https://doi.org/10.1016/S0166-3542(01)00189-9
45. Yu, M-S., Lee, J., Lee, J. M. et al. (2012). Identification of myricetin and scutellarein as novel chemical inhibitors of the SARS coronavirus helicase, nsP13. Bioorganic & Medicinal Chemistry Letters, 22(12), 4049-4054. https://doi.org/10.1016/j.bmcl.2012.04.081
46. Zhu, J., Ou, L., Zhou, Y. et al. (2020). (-)-Epigallocatechin-3-gallate induces interferon-λ2 expression to anti-influenza A virus in human bronchial epithelial cells (BEAS-2B) through p38 MAPK signaling pathway. Journal of Thoracic Disease, 12(3), 989-97. http://dx.doi.org/10.21037/jtd.2020.03.20
47. Zhu, N., Zhang, D., Wang, W. et al. (2020). A Novel Coronavirus from Patients with Pneumonia in China, 2019. The New England Journal of Medicine, 382, 727-733. https://www.nejm.org/doi/10.1056/NEJMoa2001017