Discovery Of Anti-hcv And Ns5b Rdrp Inhibition Activities Of Novel Anthraquinones By In Vitro And In-silico Approach

• ORIGINAL ARTICLE | ISSUE DETAILS: VOL NO.3 | ISSUE NO. 2

Abstract

Background: Hepatitis C virus affects over 170 million people globally. Chronic HCV infection results in major liver disorders for instance hepatocellular carcinoma and cirrhosis finally require liver transplantation. No vaccine is available yet. HCV is a tiny, encapsulated, single-stranded RNA-positive virus that is a member of the Flaviviridae family and belongs to the genus Hepacivirus. Four structural (S) and six non-structural (NS) proteins are created from its RNA genes, which encode a polyprotein precursor with about 3000 amino acids. These proteins are then digested by cellular and viral proteases. Non-structural (NS) protein NS5B, which performs a critical role in RNA-dependent RNA polymerase (RdRp), is a desired target for the development of specialised antiviral medications.

Objectives: In this research study, we studied the activities of the anthraquinone compounds, were found out the active compounds having well anti HCV and NS5B RdRp inhibition activities.

Materials & Methodology: Cassia artemisioides flowers were obtained and purified from their compounds after being harvested in Peshawar in March 2013. After being rinsed to eliminate dust, the flowers were left to air-dry for a few days. big to powdered after that. then methanol was utilized for extract in a soxhlet apparatus. The root barks of C. aretemisioides were crushed to powder the extract of the root barks of the selected plant that is soxhlet extracted andT then evaporated the solvent by using the rotary evaporator under the concentrated pressure. Column chromatography was used to separate the methanol fraction from 1, 6-dihydroxy-8-methoxy-3-methylanthraquinone(2) and 1, hydroxy-8-methoxy-methylanthraquinone (3) Prior to molecular docking, the ligand and protein were made. Then, 2D structures of the compounds were developed, and all of the compounds were then saved in 3D structures in a database. The 3D coordinates of HCV NS5B was retrieved from protein data bank .All the ligands were docked into the pocket having active residues of HCV NS5B using anti-HCV compounds were performed, The cells were produced in 96-well plates and handled with a substance at a concentration of (50 M), whereas the group in control recievwd an equivalent quantity of DMSO. Once the cells had been exposed to the compounds for 48 hours, the relative levels of the luciferase signals in the compound-treated cells compared to the DMSO controls were measured. This was done to ascertain the inhibitory effect of the compounds on HCV replication. On poly rA-U12 template primer in the presence of DMSO or the previously described medicines, HCV NS5B was inhibited by [-32P] UTP and MnCl2 as the divalent cation. Anti-HCV medications were tested for 50% inhibition at dosages of 50 M, which indicated 100% inhibition of NS5B activity in the presence of DMSO (IC50).

Results: Our findings indicate that we can also improve anthraquinones and that it may be able to create new anthraquinone analogues with enhanced cellular and enzyme-based activity that have anti-HCV and NS5B RdRp inhibitory capabilities.

Conclusion: Our research studies focused on the molecular HCV NS5B protein docked with the active residues of the anthraquinone compounds and proved that these compounds were active compounds and then treated with the HCV NS5B RdRp in vitro, Among these compounds, compound 3 is not acting as anti-HCV agent, but has good activity in the NS5B RdRp inhibition and good cell viability. Furthermore, we can also predict new anthraquinone derivatives by in silico approach and as well as can also find out their activities by in vitro process in the lab.