Changes in body weight and liver weight for control and treated Wistar female rats in the repeated dose subacute oral toxicity study (28 days)
Experimental groups
Body weight (g)
Liver (g)
Day 0
Day 7
Day 14
Day 21
Day 28
Group 1 (1 × 1 mL), n = 5
243.2
265.1
276.5
292.6
315.2
12.76
257
272.3
294
313.7
335.3
13.77
263.8
287.6
318.6
324.3
338
12.58
268.5
279.4
299.2
314.9
336.9
14.62
262.6
261
274.4
289.2
308.6
10.32
Group 2 (2 × 1 mL), n = 5
253.8
265.7
289
312.5
352
11.36
267.9
282.3
296.4
313.8
354
13.89
258
280
301.6
331.7
366.8
10.88
269
280.4
302.2
324.9
353.1
15.32
255.6
273.1
294.4
329.24
340.2
13.5
Group 3 (3 × 2 mL), n = 5
235.9
255.8
277.3
298.5
325.4
9.89
239.7
257.6
272.3
294
330.1
12.36
263.8
289.6
311.6
331.5
357
11.68
265.9
279.8
294.2
314.9
351
13.65
250.6
261.1
277.4
299.4
332.5
10.81
Control
253.1
283.6
308.6
326.9
348.9
12.54
Declarations
Acknowledgments
The authors thank Mrs. Belgin Berker Budak for her support for this research. We also thank Mrs. Begüm Buğdaycı Açıkkol, Director of Kobay Exper. Animals Inc., Turkey.
Author contributions
GGB conducted this study and wrote this paper with SÖ. MB edited this manuscript. GGB did the compound formulation and HPLC analysis. TŞ performed LC–MS–MS analysis. BM applied QTOF analysis. ŞSM did total phenolic and flavonoid content determination. CÇÇ proceeded In vitro cytotoxicity and genotoxicity tests: In vivo acute/sub-acute oral toxicity and LD50 studies. OT performed In vivo acute/sub-acute oral toxicity and LD50 studies. MB assessed the tissue samples microscopically. SÖ made Statistical analysis. All authors have read and agreed to the published version of the manuscript.
Conflicts of interest
The authors declare no conflicts of interest.
Ethical approval
Institutional Review Board statement: the animal experiments were approved by the regional Ethical Committee for animal research. The licensing ethical committee approved the experiments, including any relevant details before the animal experiments (27.11.2020/514). The authors confirm that all experiments were performed in accordance with relevant international guidelines and regulations. The authors also complied with the ARRIVE guidelines. All manipulations were made to minimize animal suffering and reduce the number of animals used.
Tu YF, Chien CS, Yarmishyn AA, Lin YY, Luo YH, Lin YT, et al. A review of SARS-CoV-2 and the ongoing clinical trials. Int J Mol Sci.2020;21:2657. [DOI] [PubMed] [PMC]
Zhou F, Yu T, Du R, Fan G, Liu Y, Liu Z, et al. Clinical course and risk factors for mortality of adult inpatients with COVID-19 in Wuhan, China: a retrospective cohort study. Lancet.2020;395:1054–62. [DOI] [PubMed] [PMC]
Lu H. Drug treatment options for the 2019-new coronavirus (2019-nCoV). Biosci Trends.2020;14:69–71. [DOI] [PubMed]
Liu X, Zhang B, Jin Z, Yang H, Rao Z. The crystal structure of COVID-19 main protease in complex with an inhibitor N3. 2020Jan26. Available from: https://www.wwpdb.org/pdb?id=pdb_00006lu7
Jin Z, Du X, Xu Y, Deng Y, Liu M, Zhao Y, et al. Structure of Mpro from SARS-CoV-2 and discovery of its inhibitors. Nature.2020;582:289–93. [DOI] [PubMed]
Xu Z, Peng C, Shi Y, Zhu Z, Mu K, Wang X, et al. Nelfinavir was predicted to be a potential inhibitor of 2019-nCov main protease by an integrative approach combining homology modelling, molecular docking and binding free energy calculation. bioRxiv [Preprint]. 2020 [cited 2021 Feb 6]. Available from https://www.biorxiv.org/content/10.1101/2020.01.27.921627v1
Wu W, Li R, Li X, He J, Jiang S, Liu S, et al. Quercetin as an antiviral agent inhibits influenza A virus (IAV) entry. Viruses.2015;8:6. [DOI] [PubMed] [PMC]
Zakaryan H, Arabyan E, Oo A, Zandi K. Flavonoids: promising natural compounds against viral infections. Arch Virol.2017;162:2539–51. [DOI] [PubMed] [PMC]
Jo S, Kim S, Shin DH, Kim MS. Inhibition of SARS-CoV 3CL protease by flavonoids. J Enzyme Inhib Med Chem.2020;35:145–51. [DOI] [PubMed] [PMC]
Kaul TN, Middleton E Jr, Ogra PL. Antiviral effectofflavonoidsonhumanviruses. J Med Virol.1985;15:71–9. [DOI] [PubMed]
Manjeet K R, Ghosh B. Quercetin inhibits LPS-induced nitric oxide and tumor necrosis factor-alpha production in murine macrophages. Int J Immunopharmacol.1999;21:435–43. [DOI] [PubMed]
Chirumbolo S. The role of quercetin, flavonols and flavones in modulating inflammatory cell function. Inflamm Allergy Drug Targets.2010;9:263–85. [DOI] [PubMed]
Liskova A, Samec M, Koklesova L, Samuel SM, Zhai K, Al-Ishaq RK, et al. Flavonoids against the SARS-CoV-2 induced inflammatory storm. Biomed Pharmacother.2021;138:111430. [DOI] [PubMed] [PMC]
Galati G, Lin A, Sultan AM, O’Brien PJ. Cellular and in vivo hepatotoxicity caused by green tea phenolic acids and catechins. Free Radic Biol Med.2006;40:570–80. [DOI] [PubMed]
Ferguson LR. Role of plant polyphenols in genomic stability. Mutat Res.2001;475:89–111. [DOI] [PubMed]
Balentine DA, Dwyer JT, Erdman JW Jr, Ferruzzi MG, Gaine PC, Harnly JM, et al. Recommendations on reporting requirements for flavonoids in research. Am J Clin Nutr.2015;101:1113–25. [DOI] [PubMed]
Slinkard K, Singleton VL. Total phenols analysis: automation and comparison with manual methods. Am J Enol Vitic.1977;28:49–55.
Panche AN, Diwan AD, Chandra SR. Flavonoids: an overview. J Nutr Sci.2016;5:e47. [DOI] [PubMed] [PMC]
Nieman DC, Mitmesser SH. Potential impact of nutrition on immune system recovery from heavy exertion: a metabolomics perspective. Nutrients.2017;9:513. [DOI] [PubMed] [PMC]
Utomo RY, Meiyanto E. Revealing the potency of citrus and galangal constituents to halt SARS-CoV-2 infection. Preprints 2020030214 [Preprint]. 2020 [cited 2021 Feb 6]. Available from: https://www.preprints.org/manuscript/202003.0214/v1
Murugan NA, Pandian CJ, Jeyakanthan J. Computational investigation on Andrographis paniculata phytochemicals to evaluate their potency against SARS-CoV-2 in comparison to known antiviral compounds in drug trials. J Biomol Struct Dyn.2021;39:4415–26. [DOI] [PubMed]
Paraiso IL, Revel JS, Stevens JF. Potential use of polyphenols in the battle against COVID-19. Curr Opin Food Sci.2020;32:149–55. [DOI] [PubMed] [PMC]
Ren Z, Yan L, Zhang N, Guo Y, Yang C, Lou Z, et al. The newly emerged SARS-like coronavirus HCoV-EMC also has an “Achilles’ heel”: current effective inhibitor targeting a 3C-like protease. Protein Cell.2013;4:248–50. [DOI] [PubMed] [PMC]
Boopathi S, Poma AB, Kolandaivel P. Novel 2019 coronavirus structure, mechanism of action, antiviral drug promises and rule out against its treatment. J Biomol Struct Dyn. 2021:39;3409–18. [DOI] [PubMed] [PMC]
Park JY, Ko JA, Kim DW, Kim YM, Kwon HJ, Jeong HJ, et al. Chalcones isolated from Angelica keiskei inhibit cysteine proteases of SARS-CoV. J Enzyme Inhib Med Chem.2016;31:23–30. [DOI] [PubMed]
Khaerunnisa S, Kurniawan H, Awaluddin R, Suhartati S, Soetjipto S. Potential inhibitor of COVID-19 main protease (Mpro) from several medicinal plant compounds by molecular docking study. Preprints [Preprint]. 2020 [cited 2021 Jun 2]. Available from: http://dx.doi.org/10.20944/preprints202003.0226.v1
Aziz N, Kim MY, Cho JY. Anti-inflammatory effects of luteolin: a review of in vitro, in vivo, and in silico studies. J Ethnopharmacol.2018;225:342–58. [DOI] [PubMed]
Smith M, Smith JC. Repurposing therapeutics for COVID-19: supercomputer-based docking to the SARS-CoV-2 viral spike protein and viral spike protein-human ACE2 interface. ChemRxiv [Preprint]. 2020 [cited 2021 Jun 2]. Available from: http://dx.doi.org/10.26434/chemrxiv.11871402.v4
Chojnacka K, Witek-Krowiak A, Skrzypczak D, Mikula K, Młynarz P. Phytochemicals containing biologically active polyphenols as an effective agent against Covid-19-inducing coronavirus. J Funct Foods.2020;73:104146. [DOI] [PubMed] [PMC]
Mhatre S, Srivastava T, Naik S, Patravale V. Antiviral activity of green tea and black tea polyphenols in prophylaxis and treatment of COVID-19: a review. Phytomedicine.2021;85:153286. [DOI] [PubMed] [PMC]
Levy E, Delvin E, Marcil V, Spahis S. Can phytotherapy with polyphenols serve as a powerful approach for the prevention and therapy tool of novel coronavirus disease 2019 (COVID-19)?Am J Physiol Endocrinol Metab. 2020;319:E689–708. [DOI] [PubMed] [PMC]
Adem S, Eyupoglu V, Sarfraz I, Rasul A, Ali M. Identification of potent COVID-19 main protease (Mpro) inhibitors from natural polyphenols: an in silico strategy unveils a hope against CORONA. Preprints 2020030333 [Preprint]. 2020 [cited 2021 Feb 4]. Available from: http://dx.doi.org/10.20944/preprints202003.0333.v1
Wong CK, Lam CW, Wu AK, Ip WK, Lee NL, Chan IH, et al. Plasma inflammatory cytokines and chemokines in severe acute respiratory syndrome. Clin Exp Immunol.2004;136:95–103. [DOI] [PubMed] [PMC]
Conti P, Ronconi G, Caraffa A, Gallenga CE, Ross R, Frydas I, et al. Induction of pro-inflammatory cytokines (IL-1 and IL-6) and lung inflammation by Coronavirus-19 (COVI-19 or SARS-CoV-2): anti-inflammatory strategies. J Biol Regul Homeost Agents.2020;34:327–31. [DOI] [PubMed]
Li Y, Yao J, Han C, Yang J, Chaudhry MT, Wang S, et al. Quercetin, inflammation and immunity. Nutrients.2016;8:167. [DOI] [PubMed] [PMC]
Scholz S, Williamson G. Interactions affecting the bioavailability of dietary polyphenols in vivo. Int J Vitam Nutr Res.2007;77:224–35. [DOI] [PubMed]
Guo Y, Mah E, Davis CG, Jalili T, Ferruzzi MG, Chun OK, et al. Dietary fat increases quercetin bioavailability in overweight adults. Mol Nutr Food Res.2013;57:896–905. [DOI] [PubMed]
Manach C, Scalbert A, Morand C, Rémésy C, Jiménez L. Polyphenols: food sources and bioavailability. Am J Clin Nutr.2004;79:727–47. [DOI] [PubMed]
de Boer VC, Dihal AA, van der Woude H, Arts IC, Wolfram S, Alink GM, et al. Tissue distribution of quercetin in rats and pigs. J Nutr.2005;135:1718–25. [DOI] [PubMed]