ANTIVIRAL ACTIVITY OF SAUSSUREA LAPPA ETHANOL EXTRACT AGAINST SARS-COV-2: IN VITRO STUDY

AVIN AINUR FITRIANINGSIH; DEWI SANTOSANINGSIH; SUSANTHY DJAJALAKSANA; SETYAWATI SOEHARTO  KARYONO; ROIHATUL MUTI'AH; MARIA INGE LUSIDA; SUMARNO RETO PRAWIRO

doi:10.22159/ijap.2024v16i2.49917

Authors

AVIN AINUR FITRIANINGSIH Doctoral Program in Medical Science, Faculty of Medicine, Universitas Brawijaya, East Java-65145, Indonesia https://orcid.org/0000-0002-3571-114X
DEWI SANTOSANINGSIH Department of Clinical Microbiology, Faculty of Medicine, Universitas Brawijaya, Malang, East Java-65145, Indonesia
SUSANTHY DJAJALAKSANA Department of Pulmonology and Respiratory Medicine, Faculty of Medicine, Universitas Brawijaya, Malang-65145, Indonesia https://orcid.org/0000-0002-2069-7357
SETYAWATI SOEHARTO KARYONO Department of Pharmacology, Faculty of Medicine, Universitas Brawijaya, Malang, East Java-65145, Indonesia
ROIHATUL MUTI'AH Department of Pharmacy, Faculty of Medicine and Health Science, UIN Maulana Malik Ibrahim, Malang, East Java-65151, Indonesia https://orcid.org/0000-0002-8196-9029
MARIA INGE LUSIDA Department of Microbiology, Faculty of Medicine, Universitas Airlangga, Surabaya, East Java-60131, Indonesia
SUMARNO RETO PRAWIRO Department of Clinical Microbiology, Faculty of Medicine, Universitas Brawijaya, Malang, East Java-65145, Indonesia

DOI:

https://doi.org/10.22159/ijap.2024v16i2.49917

Keywords:

Saussurea lappa, SARS-CoV-2, Antiviral activity

Abstract

Objective: The study aims to investigate the antiviral activity of S. lappa against Severe Acute Respiratory Syndrome-Coronavirus 2 (SARS-CoV-2) in vitro.

Methods: The extracts are obtained by ultrasonic-assisted extraction (UAE) with a 96% ethanol solvent. Thin-layer chromatography (TLC) uses n-hexane: ethyl acetate and chloroform: methanol) as mobile phases. The staining outcome is subsequently examined using UV visualizers with a wavelength of 366 nm. To assess the antiviral activity of Vero E6 cells, extracts were employed at doses of 25, 50, 75, and 100 µg/ml, with remdesivir serving as the positive control. Supernatants were collected on days 1, 2, 3, and 6 for qRT-PCR testing with target genes E and ORF1ab. Time-addition experiments were conducted to determine how the extract works as antiviral. Protein expression was tested with Western blots with antibodies S and N SARS-CoV-2.

Results: TLC identifies terpenoid chemicals present in the ethanol extract of S. lappa. The ethanol extract of S. lappa exhibited antiviral effects against SARS-CoV-2, with an inhibitory concentration 50 (IC₅₀) of 40 µg/ml, a cytotoxic concentration 50 (CC₅₀) of 131.4 µg/ml, and a selectivity index of 3.51. The extract can potentially impact the entry-post-entry phase of SARS-CoV-2 infection in Vero cells. The immunoblotting results demonstrated a reduction in the expression of S and N proteins in the treatment group compared to the negative control.

Conclusion: S. lappa ethanol extract has antiviral activity against SARS-CoV-2 based on an in vitro study.

Downloads

Download data is not yet available.

References

Vallamkondu J, John A, Wani WY, Ramadevi SP, Jella KK, Reddy PH. SARS-CoV-2 pathophysiology and assessment of coronaviruses in CNS diseases with a focus on therapeutic targets. Biochim Biophys Acta Mol Basis Dis. 2020 Oct;1866(10):165889. doi: 10.1016/j.bbadis.2020.165889, PMID 32603829.

Cucinotta D, Vanelli M. WHO declares COVID-19 a pandemic. Acta Biomed. 2020;91(1):157-60. doi: 10.23750/abm.v91i1.9397, PMID 32191675.

Harrison AG, Lin T, Wang P. Mechanisms of SARS-CoV-2 transmission and pathogenesis. Trends Immunol. 2020;41(12):1100-15. doi: 10.1016/j.it.2020.10.004, PMID 33132005.

Wang Y, Zhang D, Du G, Du R, Zhao J, Jin Y. Remdesivir in adults with severe COVID-19: a randomised, double-blind, placebo-controlled, multicentre trial. Lancet. 2020 May;395(10236):1569-78. doi: 10.1016/S0140-6736(20)31022-9, PMID 32423584.

Hassanipour S, Arab Zozani M, Amani B, Heidarzad F, Fathalipour M, Martinez-de-Hoyo R. The efficacy and safety of favipiravir in treatment of COVID-19: a systematic review and meta-analysis of clinical trials. Sci Rep. 2021 May 26;11(1):11022. doi: 10.1038/s41598-021-90551-6, PMID 34040117.

Fischer WA, Eron JJ, Holman W, Cohen MS, Fang L, Szewczyk LJ. A phase 2a clinical trial of molnupiravir in patients with COVID-19 shows accelerated SARS-CoV-2 RNA clearance and elimination of infectious virus. Sci Transl Med. 2022 Jan 19;14(628):eabl7430. doi: 10.1126/scitranslmed.abl7430, PMID 34941423.

Nikhat S, Fazil M. Overview of COVID-19; its prevention and management in the light of Unani medicine. Sci Total Environ. 2020;728:138859. doi: 10.1016/j.scitotenv.2020.138859, PMID 32334163.

Majnooni MB, Fakhri S, Bahrami G, Naseri M, Farzaei MH, Echeverría J. Alkaloids as potential phytochemicals against SARS-CoV-2: approaches to the associated pivotal mechanisms. Evid Based Complement Alternat Med. 2021;2021:6632623. doi: 10.1155/2021/6632623, PMID 34104202.

Kapoor R, Sharma B, Kanwar SS. Antiviral phytochemicals: an overview. Biochem Physiol. 2017;06(2). doi: 10.4172/2168-9652.1000220.

Da Silveira Oliveira AFC, Teixeira RR, De Oliveira AS, De Souza APM, Da Silva ML, De Paula SO. Potential antivirals: natural products targeting replication enzymes of dengue and chikungunya viruses. Molecules. 2017;22(3).

El-Far A, Shaheen H, Alsenosy A, El-Sayed Y, Al Jaouni S, Mousa S. Costus speciosus: traditional uses, phytochemistry, and therapeutic potentials. Pharmacogn Rev. 2018;12(23):1-8.

Hassan R, Masoodi MH. Saussurea lappa: a comprehensive review on its pharmacological activity and phytochemistry. Curr Tradit Med. 2020;6(1):13-23. doi: 10.2174/2215083805666190626144909.

Sukmawati S, Musfiroh I, Muchtaridi FA, Al Hindi Q. (Saussurea lappa): a narrative review of its phytochemistry and pharmacological potential against COVID-19. Int J Appl Pharm. 2022 Dec 27:1-7.

Keshavarz B, Rezaei K. Microwave- and ultrasound-assisted extraction of phenolic and flavonoid compounds from konar (Ziziphus spina-christi) fruits. Int Food Res J. 2020;27(1):47-55.

Cahyani I, Putro EW, Ridwanuloh AM, Wibowo SHB, Hariyatun H, Syahputra G. Genome profiling of SARS-CoV-2. In: Indonesia. association of southeast Asian nations, and the Neighbouring East Asian Countries: Features, Challenges, and Achievements. bioRxiv; 2021.07.06.451270. doi: 10.1101/2021.07.06.451270.

Mutiah R, Inayatin AL, Annisa R, Indrawijaya YYA, Listiyana A. Inhibition of cell cycle and induction of apoptosis y ethanol leaves extract of Chrysanthemum cinerariifolium (Trev.) in T47D breast cancer cells. Indonesian J Pharm. 2020;31(1):1-10. doi: 10.14499/indonesianjpharm31iss1pp1.

Marahatha R, Basnet S, Bhattarai BR, Budhathoki P, Aryal B, Adhikari B. Potential natural inhibitors of xanthine oxidase and HMG-CoA reductase in cholesterol regulation: in silico analysis. BMC Complement Med Ther. 2021;21(1):1. doi: 10.1186/s12906-020-03162-5, PMID 33386071.

Purniawan A, Lusida MI, Pujiyanto RW, Nastri AM, Permanasari AA, Harsono AAH. Synthesis and assessment of copper-based nanoparticles as a surface coating agent for antiviral properties against SARS-CoV-2. Sci Rep. 2022;12(1):4835. doi: 10.1038/s41598-022-08766-0, PMID 35318357.

Ohashi H, Watashi K, Saso W, Shionoya K, Iwanami S, Hirokawa T. Potential anti-COVID-19 agents, cepharanthine and nelfinavir, and their usage for combination treatment. iScience. 2021 Apr;24(4):102367. doi: 10.1016/j.isci.2021.102367, PMID 33817567.

Tang WF, Tsai HP, Chang YH, Chang TY, Hsieh CF, Lin CY. Perilla (Perilla frutescens) leaf extract inhibits SARS-CoV-2 via direct virus inactivation. Biomed J. 2021 Jun;44(3):293-303. doi: 10.1016/j.bj.2021.01.005, PMID 34119448.

Pritchetta JC, Naesens L, Montoya J. Treating HHV-6 infections: the laboratory efficacy and clinical use of anti-HHV-6 agents. In: Human herpesviruses HHV. 3rd ed. Vol. 6A, HHV-6B and HHV-7 Diagnosis and Clinical Management. Elsevier; 2014. p. 311.

Rao Vadaparthi PR, Kumar K, Sarma VU, Hussain QA, Babu KS. Estimation of costunolide and dehydrocostus lactone in Saussurea lappa and its polyherbal formulations followed by their stability studies using HPLC-DAD. Pharmacogn Mag. 2015;11(41):180-90. doi: 10.4103/0973-1296.149736, PMID 25709231.

Malhotra N, Singh M. Himalayan medicinal plants: advances in botany, production and research. Vol. 20. Academic Press; 2021. p. 202.

Quimque MTJ, Notarte KIR, de Leon VNO, Manzano JAH, Muñoz JER, Pilapil DYH. Computationally repurposed natural products targeting SARS-CoV-2 attachment and entry mechanisms. In: Frontiers of COVID-19. Berlin: Springer International Publishing; 2022. p. 505-37.

Elmaghrabi MM, Nazir T, Taha N. Saussurea costus as herbal medicines have immunomodulatory and potential nutraceutical components that may help in COVID-19 remedy. Adv J Biomed Sci. 2022;7(Jan):1-14.

Ulbegi Polat H, Serhatli M, Tas Ekiz A, Abaci I, Aksoy O, Cakirca G. Investigation of the antiviral effects of Saussurea lappa root extract against SARS-CoV-2 virus. Mol Nutr Food Res. 2023;67(14):e2200804. doi: 10.1002/mnfr.202200804, PMID 37170075.

Idriss H, Siddig B, Gonzalez-Maldonado P, Elkhair HM, Alakhras AI, Abdallah EM. Inhibitory activity of Saussurea costus extract against bacteria, candida, herpes, and SARS-CoV-2. Plants (Basel). 2023;12(3). doi: 10.3390/plants12030460, PMID 36771546.

Attallah NGM, Kabbash A, Negm WA, Elekhnawy E, Binsuwaidan R, Al-Fakhrany OM. Protective potential of Saussurea costus (Falc.) lipsch. roots against cyclophosphamide-induced pulmonary injury in rats and its in vitro antiviral effect. Pharmaceuticals (Basel). 2023 Feb 18;16(2):318. doi: 10.3390/ph16020318, PMID 37259460.

Ho YJ, Liu FC, Yeh CT, Yang CM, Lin CC, Lin TY. Micafungin is a novel anti-viral agent of chikungunya virus through multiple mechanisms. Antiviral Res. 2018 Nov;159:134-42. doi: 10.1016/j.antiviral.2018.10.005.

Idriss H, Siddig B, Maldonado PG, Elkhair HM, Alakhras AI, Abdallah EM. Phytochemical discrimination, biological activity and molecular docking of water-soluble inhibitors from Saussurea costus herb against main protease of SARS-CoV-2. Molecules. 2022 Aug 1;27(15):4908. doi: 10.3390/molecules27154908, PMID 35956858.

Khan A, Tahir Khan M, Saleem S, Junaid M, Ali A, Shujait Ali S. Structural insights into the mechanism of RNA recognition by the N-terminal RNA-binding domain of the SARS-CoV-2 nucleocapsid phosphoprotein. Comput Struct Biotechnol J. 2020;18:2174-84. doi: 10.1016/j.csbj.2020.08.006, PMID 32837710.

Poleboyina PK, Pawar SC. Comparative analysis of small molecules and natural plant compounds as therapeutic inhibitors targeting rdrp and nucleocapsid proteins of sars cov 2: an in silico approach. Asian J Pharm Clin Res. 2023;16(10):208-28. doi: 10.22159/ajpcr.2023.v16i10.48095.

Chang CK, Hou MH, Chang CF, Hsiao CD, Huang TH. The SARS coronavirus nucleocapsid protein forms and functions. Antiviral Res. 2014;103(1):39-50. doi: 10.1016/j.antiviral.2013.12.009, PMID 24418573.