Cyclopeptide Kalata B12 as HCV-NS5A potent Inhibitor: Cyclopeptide Kalata B12 as HCV-NS5A Potent Inhibitor

Faiza  Shams; Nazia  Kanwal; Somayya  Tariq; Ayesha  Malik; Kausar  Malik; Bushra  Ijaz

doi:10.54393/pbmj.v5i5.483

Authors

Faiza Shams Centre of Excellence in Molecular Biology, University of the Punjab Lahore, Pakistan
Nazia Kanwal Superior University Lahore, Pakistan
Somayya Tariq Centre of Excellence in Molecular Biology, University of the Punjab Lahore, Pakistan
Ayesha Malik Centre of Excellence in Molecular Biology, University of the Punjab Lahore, Pakistan
Kausar Malik Centre of Excellence in Molecular Biology, University of the Punjab Lahore, Pakistan
Bushra Ijaz Centre of Excellence in Molecular Biology, University of the Punjab Lahore, Pakistan

DOI:

https://doi.org/10.54393/pbmj.v5i5.483

Keywords:

HCV, Cyclopeptide, NS5A, Inhibition, Antiviral

Abstract

Hepatitis C Virus (HCV) is the leading cause of liver diseases globally, causing severe complications such as liver fibrosis, cirrhosis, and hepatocellular carcinoma (HCC). Despite the advent of successful regimens, still, 71 million individuals are chronically infected every year. Therefore, more accessible novel therapies are needed to fight the challenges such as adverse effects, genotype selectivity, and resistance to these regimens due to viral mutations. HCV NS5A is a non-structural phosphoprotein, with its pivotal role in viral replication assembly, and has been the target of continuous research. Cyclopeptides are an emerging class of peptides reported to have antiviral, anticancer, and antimicrobial properties. These cyclopeptides have exceptional resistance to thermal, chemical, or enzymatic degradation. Herein, we present the inhibitory potential of cyclopeptide Kalata B12 against the HCV NS5A gene. Objective: To investigate the antiviral potential of Kalata B2, Kalata B12, and cycloviolacin O14 against HCV NS5A Methods: We investigated thirty cyclopeptides through molecular docking analysis for their anti-HCV-NS5A inhibition potential. Three cyclopeptides, Kalata B2, Kalata B12, and cycloviolacin O14 showed minimum binding energies, for their antiviral potential. The defense-related, circular mini-protein Kalata B12 showed an impressive docking score of -9.80 Kcal/mol. Further, it was synthesized and went through cytotoxicity analysis via MTT assay on HepG2 cell line, which showed more than 85% cell viability at submicromolar concentrations. Results: The peptide Kalata B12 showed significant (***P<0.0001) inhibition of NS5A gene (approx. 75%) at 100nM in In vitro trials, confirmed by real-Time PCR analysis. Conclusions: Kalata B12 cyclopeptide was found to be a potential HCV NS5A inhibitor

References

Gao M, Nettles RE, Belema M, Snyder LB, Nguyen VN, Fridell RA et al. Chemical genetics strategy identifies an HCV NS5A inhibitor with a potent clinical effect. Nature. 2010 May 6;465(7294):96-100. doi: 10.1038/nature08960.

Jardim ACG, Shimizu JF, Rahal P, Harris M. Plant-derived antivirals against hepatitis c virus infection. Virol J. 2018 Feb 13;15(1):34. doi: 10.1186/s12985-018-0945-3.

Reddy BU, Mullick R, Kumar A, Sharma G, Bag P, Roy CL et al. A natural small molecule inhibitor corilagin blocks HCV replication and modulates oxidative stress to reduce liver damage. Antiviral Res. 2018 Feb;150:47-59. doi: 10.1016/j.antiviral.2017.12.004.

Chan J, Gogela N, Zheng H, Lammert S, Ajayi T, Fricker Z et al. Direct-Acting Antiviral Therapy for Chronic HCV Infection Results in Liver Stiffness Regression Over 12 Months Post-treatment. Dig Dis Sci. 2018 Feb;63(2):486-492. doi: 10.1007/s10620-017-4749-x.

Pawlotsky JM. New hepatitis C therapies: the toolbox, strategies, and challenges. Gastroenterology. 2014 May;146(5):1176-92. doi: 10.1053/j.gastro.2014.03.003.

Pawlotsky JM. New hepatitis C virus (HCV) drugs and the hope for a cure: concepts in anti-HCV drug development. Semin Liver Dis. 2014 Feb;34(1):22-9. doi: 10.1055/s-0034-1371007.

Kotwal GJ. Natural antivirals against human viruses. Virol. Mycol. 2014;3:e107.

Ganta KK, Mandal A, Debnath S, Hazra B, Chaubey B. Anti‐HCV activity from semi‐purified methanolic root extracts of Valeriana wallichii. Phytotherapy research. 2017 Mar;31(3):433-40. doi.org/10.1002/ptr.5765.

Lemm JA, O'Boyle D 2nd, Liu M, Nower PT, Colonno R, Deshpande MS et al. Identification of hepatitis C virus NS5A inhibitors. J Virol. 2010 Jan;84(1):482-91. doi: 10.1128/JVI.01360-09.

Gao M, O'Boyle DR 2nd, Roberts S. HCV NS5A replication complex inhibitors. Curr Opin Pharmacol. 2016 Oct;30:151-157. doi: 10.1016/j.coph.2016.07.014.

Badillo A, Receveur-Brechot V, Sarrazin S, Cantrelle FX, Delolme F, Fogeron ML et al. Overall Structural Model of NS5A Protein from Hepatitis C Virus and Modulation by Mutations Confering Resistance of Virus Replication to Cyclosporin A. Biochemistry. 2017 Jun 20;56(24):3029-3048. doi: 10.1021/acs.biochem.7b00212.

Ireland DC, Colgrave ML, Craik DJ. A novel suite of cyclotides from Viola odorata: sequence variation and the implications for structure, function and stability. Biochem J. 2006 Nov 15;400(1):1-12. doi: 10.1042/BJ20060627.

Troeira Henriques S, Huang YH, Chaousis S, Wang CK, Craik DJ. Anticancer and toxic properties of cyclotides are dependent on phosphatidylethanolamine phospholipid targeting. Chembiochem. 2014 Sep 5;15(13):1956-65. doi: 10.1002/cbic.201402144.

Khalil R, Ashraf S, Khalid A, Ul-Haq Z. Exploring Novel N-Myristoyltransferase Inhibitors: A Molecular Dynamics Simulation Approach. ACS Omega. 2019 Aug 15;4(9):13658-13670. doi: 10.1021/acsomega.9b00843.

Schmittgen TD, Livak KJ. Analyzing real-time PCR data by the comparative C(T) method. Nat Protoc. 2008;3(6):1101-8. doi: 10.1038/nprot.2008.73.

Sanghani HV, Ganatra SH, Pande R. Molecular—docking studies of potent anticancer agent. J Comput Sci Syst Biol. 2012;5(1):12-5.

Gao M. Antiviral activity and resistance of HCV NS5A replication complex inhibitors. Curr Opin Virol. 2013 Oct;3(5):514-20. doi: 10.1016/j.coviro.2013.06.014.

Donato MT, Tolosa L, Gómez-Lechón MJ. Culture and Functional Characterization of Human Hepatoma HepG2 Cells. Methods Mol Biol. 2015;1250:77-93. doi: 10.1007/978-1-4939-2074-7_5.

Marks DC, Belov L, Davey MW, Davey RA, Kidman AD. The MTT cell viability assay for cytotoxicity testing in multidrug-resistant human leukemic cells. Leuk Res. 1992 Dec;16(12):1165-73. doi: 10.1016/0145-2126(92)90114-m.

Rajik M, Jahanshiri F, Omar AR, Ideris A, Hassan SS, Yusoff K. Identification and characterisation of a novel anti-viral peptide against avian influenza virus H9N2. Virol J. 2009 Jun 5;6:74. doi: 10.1186/1743-422X-6-74.

Lee BW, Quy Ha TK, Park EJ, Cho HM, Ryu B, Doan TP et al. Melicopteline A-E, Unusual Cyclopeptide Alkaloids with Antiviral Activity against Influenza A Virus from Melicope pteleifolia. J Org Chem. 2021 Jan 15;86(2):1437-1447. doi: 10.1021/acs.joc.0c02137.