EC Paediatrics

Background: A novel coronavirus pneumonia (COVID-19) outbreak in Wuhan, China, was caused by the severe acute respiratory syndrome corona virus 2 (SARS-CoV-2), and has now expanded to over 200 countries and territories, infecting more than 2 million people in US and caused over 20 thousand deaths worldwide.

Kumazasa derivatives are reported safety drug to improve anti microorganisms including coronavirus in Japan and its main active ingredient is tricin (4,5,7-trihydroxy-3,5-dimethoxyflavon). The Kumazasa derivatives can suppress the human cytomegalovirus (HCMV) of the DNA virus and the coronavirus pneumonia virus of RNA virus replication in vitro.

Materials and Methods: Human embryonic pulmonary fibroblast (HEL), obtained from human fetal lungs cultivated in 9.6 cm² shell until confluence with Dulbecco’s Modified Eagle Medium (DMEM) medium was carried out, supplemented with 8% amount of fetal bovine serum (FCS). Towne strain of HCMV has been adsorbed and infected in a variety of infections (MOI) = 1 in HEL cells. After incubation at 37°C for 1 hour, concentrations of 0.1, 1μ, 10 μM of tricin or novel compounds were added and cultivated for 6 days, and infectious HCMV numbers in the culture supernatant were quantified by the plaque method. In the plaque method, HEL cells cultivated to flow into a 24-wells plate were adsorbed on a culture supernatant that was diluted for 1 hour at 37°C 1 hour and then superimposed with DMEM medium with 2% FCS and 0.4% agar for cultivation. After several days (6 to 12 days), the number of plaques displayed under the microscope was counted and the viral load measured.

Results: Comparison of the antiviral effect of GCV, Tricin, Anti-HCMV effects were investigated for several novel compounds found by docking simulation. Here the results that show the most significant effect are shown. The novel compound has been found to exhibit a stronger anti-HCMV effect compared to GCV and Tricin. Next EC-50 values were determined and compared.

Conclusion: The network pharmacological strategy integrated molecular docking to investigate the mechanism of action of AHI against COVID-19. It provides protein targets associated with COVID-19 that can be further tested as therapeutic targets of this safety ingredients.

Keywords: Anti-Virus New CAM; Anti-Cytomegalovirus Agent; CDK9; Tricin; Corona Virus

1. Spungen AM., et al. "Pulmonary obstruction in persons with cervical spine lesions exposed by bronchodilator administration". Paraplegia6 (1993): 404-407.
2. Yamaguchi N., et al "Aspect of QOL Assessment and Proposed New Scale for Evaluation". Open Journal of Immunology 5 (2015): 147-182.
3. Kishida K., et al. "Cranial Irradiation and Lymphocyte Subpopulation in Acute Lymphatic Leukemia". Journal of Pediatrics5 (1978): 785-786.
4. Murgita RA and Tomasi Jr TB. "Suppression of the Immune Response by Alpha-Fetoprotein". The Journal of Experimental Medicine2 (1975): 269-286.
5. Koshimo H., et al. "Maternal Antigenic Stimulation Actively Produces Suppressor Activity in Offspring". Developmental and Comparative Immunology1 (1989): 79-85.
6. Zoeller M. "Tolerization during Pregnancy: Impact on the Development of Antigen-Specific Help and Suppression". European Journal of Immunology12 (1988): 1937-1943.
7. Auerback R and Clark S. "Immunological Tolerance: Transmission from Mother to Offspring". Science 4205 (1975): 811-813.
8. Shinka S., et al. "Immunological responsiveness in mice. I. Immunological responsiveness induced in embryonic mice by maternofetal transfer of human globulin". Biken Journal2 (1974): 59-72.
9. Aase J M., et al. "Mumps virus infection in pregnant women and the immunological response of their offspring". The New England Journal of Medicine26 (1972): 1379-1382.
10. Cramer DV., et al. "Immunologic Sensitization Before Birth". American Journal of Obstetrics and Gynecology3 (1974): 431-439.
11. Stoika RS., et al. "In Vitro Studies of Activation of Phagocytic Cells by Bioactive Peptides". Journal of Physiology and Pharmacology 4 (2002): 675-688.
12. Paul G., et al. "CD4+ but not CD8+ T cells are required for oral tolerance induction". International Immunology3 (1995): 501-504.
13. Yamaguchi N., et al. "Rodent Macrophage Select Vin Blank Together with Vin Rouge According to SO Level in Situ". Open Journal of Rheumatology and Autoimmune Diseases4 (2014): 240-247.
14. Yamaguchi N., et al. "Overall Estimation of Anti-Oxidant Activity by Mammal Macrophage". Open Journal of Rheumatology and Autoimmune Diseases 4 (2014): 13-21.
15. Elbim C., et al. "The Role of Phagocytes in HIV-Related Oxidative Stress". Journal of Clinical Virology3 (2001): 99-109.
16. Asumi Sakai., et al. “Anti-human cytomegalovirus activity of components from Sasa albo-marginata (Kumazasa in Japan)”. Antiviral Chemistry and Chemotherapy 3 (2008): 125-132.
17. Kurumi Yazawa., et al. “Anti-Coronavirus Pneumonia Virus Activity of Tricin, 4',5,7-Trihydroxy3',5'-dioxy”. Antiviral Chemistry and Chemotherapy 1 (2011): 1-11.
18. Kazuhiko Akuzawa., et al. “Inhibiting effect of tricine derivative from Sasa albo-marginata on the replication of the human cytomegalovirus”. Antiviral Research3 (2011): 296-303.
19. T Murayama., et al. “Anti-cytomegalovirus effects of tricin are dependent on CXCL11”. Microbe Infection12 (2012): 1086-1092.
20. Tsugiya Murayama., et al. “Human cytomegalovirus replication supported by virus-induced activation of CCL2-CCR2 interactions”. Biochemical and Biophysical Research Communications 3 (2014): 321-325.
21. Rie Yamada., et al. “Synergistic effects of combination treatment with ganciclovir and tricin on human cytomegalovirus replication in vitro”. Antiviral Research 125 (2016): 79-83.
22. Yumiko Akai., et al. “Inhibition of human cytomegalovirus replication by tricin is associated with depressive CCL2 expression”. Antiviral Research 148 (2017): 15-19.
23. Hidetaka Sadanari., et al. “The antihuman cytomegalovirus tricin inhibits cyclin-dependent kinase 9”. FEBS Open Bio4 (2018): 646-654.
24. Akimasa Itho., et al. “Tricin inhibits CCL5 induction required for the efficient growth of human cytomegalovirus”. Microbiology and Immunology 5 (2018): 341-347.
25. Kazuhiro J Fujimoto., et al. “A silico-designated silico in silico flavone derivative, 6-fluoro-4-hydroxy-3,5-dimethoxyflavon has a greater antihuman cytomegalovirus effect”. Antiviral Research 154 (2018): 10-16.