Fig 1.
Chromatograms of GC-MS analysis of EGYVIR extract.
Table 1.
Phyto-constituents present in EGYVIR extract as identified by GC-MS analysis.
Fig 2.
EGYVIR extract cytotoxicity and viral inhibition activity on Vero-E6 cell lines.
(A and B) The cytotoxicity and antiviral activity of EGYVIR on SARS-COV-2 based on the dose response (0.4 to 0.05 μg/ml) was determined using MTT and plaque reduction assay, respectively. The result showed that the 50% cytotoxic concentration (CC50) was 0.57 μg/ml. The concentrations from 0.4 to 0.1 μg/ml showed 92% to 88% SARS -CoV-2 inhibition. At 0.05 μg/ml of EGYVIR, the viral inhibition decreased to 54.6%. (C and D) the cytotoxicity and antiviral activity of hydroxychloroquine as a reference drug, respectively.
Fig 3.
The inhibitory effect of EGYVIR extract (Extract) on Vero cells vs hydroxychloroquine (HCQ).
Fig 4.
Mode of action of EGYVIR on Vero E6 cells.
Fig 5.
Protein levels of pro-inflammatory cytokines IL-6, TNFα, and NF-κB.
(A) HUH7 cells treated with EGYVIR significantly down regulates IκBα levels at studied time point compared with SARS-COV-2 infected cells. (B) HUH7 cells treated with EGYVIR significantly down regulates TNFα levels at studied time point compared with SARS-COV-2 infected cells. (C) HUH7 cells treated with EGYVIR significantly down regulates IL-6 levels at studied time point compared with SARS-COV-2 infected cells. (D) EGYVIR significantly attenuates the nuclear translocation of p50 subunit in HUH7 cells compared with the SARS-COV-2 infected cells where the nuclear translocation became obvious after 2h post infection and significantly stable for 24h post infection.
Fig 6.
Transcriptional levels of pro-inflammatory cytokines Iκβα, TNFα, and IL-6.
(A) EGYVIR down regulates mRNA levels of total Ikβα that affected the proteosomal degradation of the Ikβα that attenuates the NF-κB nuclear translocation. (B) EGYVIR down regulates the log fold change of TNFα induced by SARS- COV-2 infection by 2–3 times at the studied time points. (C) EGYVIR down regulates the log fold change of TNFα induced by SARS-COV-2 infection by 2–3 times at the studied time points.
Fig 7.
Molecular docking of EGYVIR ingredients on RBD of SARS-COV-2.
Fig 8.
Molecular docking of EGYVIR ingredients on NFkB p50 subunit.
Table 2.
Showed chemical structure and molecular docking of ingredients of EGYVIR with viral spike RBD, and cellular P50 subunit.
Fig 9.
EGYVIR molecular mechanism as a virucidal and cytokine storm disturbance via NF-Kb pathway.
EGYVIR works as a virucidal agent on SARS-COV-2 targeting the spike protein which prevent interaction with host cell receptor. Our docking studies showed that there are good bind affinities between EGYVIR extract active ingredients and RBD of human ACE2 receptor which disturbs the attachment of SARS-COV-2 virus with host, also our docking studies showed a good binding affinity between EGYVIR active ingredients and p50 subunit of NF-κB which attenuates NF-κB pathways. Our lab studies showed that EGYVIR inhibits the nuclear translocation of p50 and disturbs NF-κB pathway resulting in decrease of cytokine storm by down-regulating IL-6 and TNFα production.