Antiviral activity of different misai kucing extracts against herpes simplex virus type 1


Herpes is a challenging opportunistic infection caused by herpes simplex virus 1 (HSV-1). Emergence of drug resistance is the worst catastrophe that encountered the dedicated efforts to eradicate the disease. This urged the scientists to search for other alternatives from the natural products sanctuary. In this study, aqueous, methanol and ethanol extracts of Orthosiphon stamineus leaves (ALE, MLE & ELE) were prepared and their yield extract and phytochemical contents were determined. Their anti HSV-1 effect was screened using plaque reduction assay and their safety limit was assessed after determining their cytotoxicity against Vero cells using MTT assay. This was followed by mechanism elucidation study which included pre-treatment, attachment, virucidal, penetration, virus yield and time removal and time addition assays. MLE got the highest yield extract and phytochemical content as compared to the others. All of them were considered as non-toxic to Vero cells as their (CC50s > 0.2 mg/ml). Furthermore, their anti-HSV-1 effect shows that all of them had a selective effect in the following order (MLE>ELE>ALE. Their anti-HSV-1 effect was ascribed to their direct virucidal effect or interference with the viral entry through modifying the viral ligand molecules or preventing fusion of the viral envelop with the host cell membrane. In this regard, MLE produced the most powerful effect and it was the only which inhibited the intracellular activity of the virus at all the replication stages. Overall, O. stamineus leaves can be used to develop new antiherpetic agent.


  • Arthanari SK, Vanitha J, Ganesh M, Venkateshwaran K, Clercq D (2012) Evaluation of antiviral and cytotoxic activities of methanolic extract of S. grandiflora (Fabaceae) flowers. Asian Pacific Journal of Tropical Biomedicine 2(2): S855-S858.
  • Atanasiu D, Whitbeck JC, Cairns TM, Reilly B, Cohen GH, Eisenberg RJ (2007) Bimolecular complementation reveals that glycoproteins gB and gH/gL of herpes simplex virus interact with each other during cell fusion. Proceedings of the National Academy of Sciences 104(47): 18718-18723.
  • Barnard DL, Huffman JH, Morris JL, Wood SG, Hughes BG, Sidwell RW (1992) Evaluation of the antiviral activity of anthraquinones, anthrones and anthraquinone derivatives against human cytomegalovirus. Antiviral research 17(1): 63-77.
  • Campadelli‐Fiume G, Amasio M, Avitabile E, Cerretani A, Forghieri C, Gianni T, Menotti L (2007) The multipartite system that mediates entry of herpes simplex virus into the cell. Reviews in medical virology 17(5): 313-326.
  • Cassidy-Stone A, Chipuk JE, Ingerman E, Song C, Yoo C, Kuwana T, Kurth MJ, Shaw JT, Hinshaw JE, Green DR (2008) Chemical inhibition of the mitochondrial division dynamin reveals its role in Bax/Bak-dependent mitochondrial outer membrane permeabilization. Developmental cell 14(2): 193-204.
  • Cheng H-Y, Lin C-C, Lin T-C (2002) Antiviral properties of prodelphinidin B-2 3′-O-gallate from green tea leaf. Antiviral Chemistry and Chemotherapy 13(4): 223-229.
  • Cheng H-Y, Lin T-C, Yang C-M, Wang K-C, Lin L-T, Lin C-C (2004) Putranjivain A from Euphorbia jolkini inhibits both virus entry and late stage replication of herpes simplex virus type 2 in vitro. Journal of Antimicrobial Chemotherapy 53(4): 577-583.
  • Côté M, Misasi J, Ren T, Bruchez A, Lee K, Filone CM, Hensley L, Li Q, Ory D, Chandran K (2011). Small molecule inhibitors reveal Niemann-Pick C1 is essential for Ebola virus infection. Nature 477(7364): 344-348.
  • Danaher RJ, Wang C, Dai J, Mumper RJ, Miller CS (2011) Antiviral effects of blackberry extract against herpes simplex virus type 1. Oral Surgery, Oral Medicine, Oral Pathology, Oral Radiology, and Endodontology 112(3): e31-e35.
  • De Logu A, Loy G, Pellerano ML, Bonsignore L, Schivo ML (2000) Inactivation of HSV-1 and HSV-2 and prevention of cell-to-cell virus spread by Santolina insularis essential oil. Antiviral research 48(3): 177-185.
  • Fadeyi SA, Fadeyi OO, Adejumo AA, Okoro C, Myles EL (2013) In vitro anticancer screening of 24 locally used Nigerian medicinal plants. BMC complementary and alternative medicine 13(1): 1.
  • Franquet T, Lee KS, Müller NL (2003) Thin-section CT findings in 32 immunocompromised patients with cytomegalovirus pneumonia who do not have AIDS. American Journal of Roentgenology 181(4): 1059-1063.
  • Garner JA (2003) Herpes simplex virion entry into and intracellular transport within mammalian cells. Advanced drug delivery reviews 55(11): 1497-1513.
  • Grossmann JG, Sharff AJ, O’Hare P, Luisi B (2001) Molecular shapes of transcription factors TFIIB and VP16 in solution: implications for recognition. Biochemistry 40(21): 6267-6274.
  • Harden EA, Falshaw R, Carnachan SM, Kern ER, Prichard MN (2009) Virucidal activity of polysaccharide extracts from four algal species against herpes simplex virus. Antiviral research 83(3): 282-289.
  • Hierholzer J, Killington R (1996) Virus isolation and quantitation. Virology methods manual: 25-46.
  • Hong-juan J, Yan-jie Y, Guo-qiang Z (2017) Hydrogen Peroxide Involved in Tip Growth of Wheat (TriticumAestivum) Root Hairs by High Concentration Chlorogenic Acid. Canadian Journal of Agriculture and Crops, 2(1): 22-33.
  • Jaime V, Florencia M, Campos RH, Martino VS, Cavallaro LV, Muschietti LV (2013) Antipoliovirus activity of the organic extract of Eupatorium buniifolium: isolation of Euparin as an active compound. Evidence-Based Complementary and Alternative Medicine.
  • Kajino K, Jilbert AR, Saputelli J, Aldrich CE, Cullen J, Mason WS (1994) Woodchuck hepatitis virus infections: very rapid recovery after a prolonged viremia and infection of virtually every hepatocyte. Journal of virology 68(9): 5792-5803.
  • Kratz JM, Andrighetti-Fröhner CR, Kolling DJ, Leal PC, Cirne-Santos CC, Yunes RA, ... Barardi CRM (2008) Anti-HSV-1 and anti-HIV-1 activity of gallic acid and pentyl gallate. Memorias do Instituto Oswaldo Cruz 103(5): 437-442.
  • Monavari SH, Shahrabadi MS, Keyvani H, Bokharaei-Salim F (2012) Evaluation of in vitro antiviral activity of Chelidonium majus L. against herpes simplex virus type-1. African Journal of Microbiology Research 6(20): 4360-4364.
  • N’diaye AD, Kankou MSA (2020) Sorption of Aspirin from Aqueous Solutions using Rice Husk as Low Cost Sorbent. Journal of Environmental Treatment Techniques, 8(1): 1-5.
  • Naithani R, Huma LC, Holland LE, Shukla D, McCormick DL, Mehta RG, Moriarty RM (2008) Antiviral activity of phytochemicals: a comprehensive review. Mini reviews in medicinal chemistry 8(11): 1106-1133.
  • Prichard MN, Turk SR, Coleman LA, Engelhardt SL, Shipman Jr C, Drach JC (1990) A microtiter virus yield reduction assay for the evaluation of antiviral compounds against human cytomegalovirus and herpes simplex virus. Journal of virological methods 28(1): 101-106.
  • Pujol CA, Estevez JM, Carlucci MJ, Ciancia M, Cerezo AS, Damonte EB (2002) Novel DL-galactan hybrids from the red seaweed Gymnogongrus torulosus are potent inhibitors of herpes simplex virus and dengue virus. Antiviral Chemistry and Chemotherapy 13(2): 83-89.
  • Ritchie RJ (2006) Consistent sets of spectrophotometric chlorophyll equations for acetone, methanol and ethanol solvents. Photosynthesis research 89(1): 27-41.
  • Saddi M, Sanna A, Cottiglia F, Chisu L, Casu L, Bonsignore L, De Logu A (2007) Antiherpevirus activity of Artemisia arborescens essential oil and inhibition of lateral diffusion in Vero cells. Annals of Clinical Microbiology and Antimicrobials 6(1): 10.
  • Schuhmacher A, Reichling J, Schnitzler P (2003) Virucidal effect of peppermint oil on the enveloped viruses herpes simplex virus type 1 and type 2 in vitro. Phytomedicine 10(6-7): 504-510.
  • Shogan B, Kruse L, Mulamba GB, Hu A, Coen DM (2006) Virucidal activity of a GT-rich oligonucleotide against herpes simplex virus mediated by glycoprotein B. Journal of virology 80(10): 4740-4747.
  • Souza TML, De Souza MCBV, Ferreira VF, Canuto CVBS, Marques IP, Fontes CFL, Frugulhetti, I. C. (2008). Inhibition of HSV-1 replication and HSV DNA polymerase by the chloroxoquinolinic ribonucleoside 6-chloro-1, 4-dihydro-4-oxo-1-(β-d-ribofuranosyl) quinoline-3-carboxylic acid and its aglycone. Antiviral research, 77(1), 20-27.
  • Sultana B, Anwar F, Ashraf M (2009) Effect of extraction solvent/technique on the antioxidant activity of selected medicinal plant extracts. Molecules 14(6): 2167-2180.
  • Sydiskis RJ, Owen DG, Lohr JL, Rosler KH, Blomster RN (1991) Inactivation of enveloped viruses by anthraquinones extracted from plants. Antimicrobial agents and chemotherapy 35(12): 2463-2466.
  • Taguri T, Tanaka T, Kouno I (2004) Antimicrobial activity of 10 different plant polyphenols against bacteria causing food-borne disease. Biological and Pharmaceutical Bulletin 27(12): 1965-1969.
  • Torky ZA (2013) Antiviral activity of polyphenols extracts from Daucus carota against Herpes simplex virus type 1. TOJSAT: The Online J Sci Technol 2013 3 (1): 20 32.
  • Whitbeck JC, Peng C, Lou H, Xu R, Willis SH, De Leon MP, ... Soulika AM (1997) Glycoprotein D of herpes simplex virus (HSV) binds directly to HVEM, a member of the tumor necrosis factor receptor superfamily and a mediator of HSV entry. Journal of virology 71(8): 6083-6093.
  • Xiang Y, Pei Y, Qu C, Lai Z, Ren Z, Yang K, ... Liu Q (2011) In vitro Anti‐Herpes Simplex Virus Activity of 1, 2, 4, 6‐Tetra‐O‐galloyl‐β‐d‐glucose from Phyllanthus emblica L.(Euphorbiaceae). Phytotherapy Research 25(7): 975-982.
  • Xiong HR, Luo J, Hou W, Xiao H, Yang ZQ (2011) The effect of emodin, an anthraquinone derivative extracted from the roots of Rheum tanguticum, against herpes simplex virus in vitro and in vivo. Journal of ethnopharmacology 133(2): 718-723.
  • Yucharoen R, Anuchapreeda S, Tragoolpua Y (2011) Anti-herpes simplex virus activity of extracts from the culinary herbs Ocimum sanctum L., Ocimum basilicum L. and Ocimum americanum L. African Journal of Biotechnology 10(5): 860-866.
  • Zeedan GSG, El-Razik KhAE-H A, Abdel-Shafy S, Farag TK, Mahmoud AH (2019) The Effects of Green Tea and Propolis Extracts on pro-inflammatory cytokines TNF-α, IFN-γ, IL2, and Immunoglobulin Production in Experimentally Infected Rabbits with Bovine Herpesvirus-1. World Vet. J 9(4): 329-339. DOI:


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