{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,10]],"date-time":"2026-09-10T22:56:44Z","timestamp":1789081004405,"version":"build-2803163510"},"reference-count":56,"publisher":"MDPI AG","issue":"2","license":[{"start":{"date-parts":[[2022,1,14]],"date-time":"2022-01-14T00:00:00Z","timestamp":1642118400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Viral infections represent a serious threat to the world population and are becoming more frequent. The search and identification of broad-spectrum antiviral molecules is necessary to ensure new therapeutic options, since there is a limited availability of effective antiviral drugs able to eradicate viral infections, and consequently due to the increase of strains that are resistant to the most used drugs. Recently, several studies on antimicrobial peptides identified them as promising antiviral agents. In detail, amphibian skin secretions serve as a rich source of natural antimicrobial peptides. Their antibacterial and antifungal activities have been widely reported, but their exploitation as potential antiviral agents have yet to be fully investigated. In the present study, the antiviral activity of the peptide derived from the secretion of Rana tagoi, named AR-23, was evaluated against both DNA and RNA viruses, with or without envelope. Different assays were performed to identify in which step of the infectious cycle the peptide could act. AR-23 exhibited a greater inhibitory activity in the early stages of infection against both DNA (HSV-1) and RNA (MeV, HPIV-2, HCoV-229E, and SARS-CoV-2) enveloped viruses and, on the contrary, it was inactive against naked viruses (PV-1). Altogether, the results indicated AR-23 as a peptide with potential therapeutic effects against a wide variety of human viruses.<\/jats:p>","DOI":"10.3390\/ijms23020883","type":"journal-article","created":{"date-parts":[[2022,1,14]],"date-time":"2022-01-14T12:34:04Z","timestamp":1642163644000},"page":"883","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":50,"title":["The Broad-Spectrum Antiviral Potential of the Amphibian Peptide AR-23"],"prefix":"10.3390","volume":"23","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-1648-4519","authenticated-orcid":false,"given":"Annalisa","family":"Chianese","sequence":"first","affiliation":[{"name":"Department of Experimental Medicine, University of Campania \u201cLuigi Vanvitelli\u201d, 80138 Naples, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7991-8700","authenticated-orcid":false,"given":"Carla","family":"Zannella","sequence":"additional","affiliation":[{"name":"Department of Experimental Medicine, University of Campania \u201cLuigi Vanvitelli\u201d, 80138 Naples, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alessandra","family":"Monti","sequence":"additional","affiliation":[{"name":"Institute of Biostructures and Bioimaging (IBB), National Research Council (CNR), 80134 Naples, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0395-7962","authenticated-orcid":false,"given":"Anna","family":"De Filippis","sequence":"additional","affiliation":[{"name":"Department of Experimental Medicine, University of Campania \u201cLuigi Vanvitelli\u201d, 80138 Naples, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nunzianna","family":"Doti","sequence":"additional","affiliation":[{"name":"Institute of Biostructures and Bioimaging (IBB), National Research Council (CNR), 80134 Naples, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3321-4331","authenticated-orcid":false,"given":"Gianluigi","family":"Franci","sequence":"additional","affiliation":[{"name":"Department of Medicine, Surgery and Dentistry \u201cScuola Medica Salernitana\u201d, University of Salerno, 84081 Baronissi, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1576-6290","authenticated-orcid":false,"given":"Massimiliano","family":"Galdiero","sequence":"additional","affiliation":[{"name":"Department of Experimental Medicine, University of Campania \u201cLuigi Vanvitelli\u201d, 80138 Naples, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2022,1,14]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"105924","DOI":"10.1016\/j.ijantimicag.2020.105924","article-title":"Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and coronavirus disease-2019 (COVID-19): The epidemic and the challenges","volume":"55","author":"Lai","year":"2020","journal-title":"Int. J. Antimicrob. Agents"},{"key":"ref_2","unstructured":"WHO (2021, December 09). Coronavirus Disease (COVID-19) Pandemic. Available online: https:\/\/www.who.int\/emergencies\/diseases\/novel-coronavirus-2019."},{"key":"ref_3","unstructured":"WHO (2021, December 09). Coronavirus (COVID-19) Dashboard. Available online: https:\/\/covid19.who.int\/."},{"key":"ref_4","first-page":"e21358","article-title":"Temporin G, an amphibian antimicrobial peptide against influenza and parainfluenza respiratory viruses: Insights into biological activity and mechanism of action","volume":"35","author":"Casciaro","year":"2021","journal-title":"FASEB J."},{"key":"ref_5","unstructured":"WHO (2021, December 09). Influenza (Seasonal). Available online: https:\/\/www.who.int\/news-room\/fact-sheets\/detail\/influenza-(seasonal)."},{"key":"ref_6","unstructured":"CDC (2021, December 09). Parainfluenza Types 1\u20133 National Trends, Available online: https:\/\/www.cdc.gov\/surveillance\/nrevss\/human-paraflu\/natl-trend.html."},{"key":"ref_7","doi-asserted-by":"crossref","unstructured":"Saxena, S.K., Mishra, N., and Saxena, R. (2009). Advances in antiviral drug discovery and development: Part II: Advancements in antiviral drug development. Future Virol., 43.","DOI":"10.2217\/fvl.09.1"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"5","DOI":"10.1007\/s10096-019-03674-0","article-title":"Melittin: A venom-derived peptide with promising anti-viral properties","volume":"39","author":"Memariani","year":"2020","journal-title":"Eur. J. Clin. Microbiol. Infect. Dis."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1038\/ijos.2016.3","article-title":"New strategies against drug resistance to herpes simplex virus","volume":"8","author":"Jiang","year":"2016","journal-title":"Int. J. Oral Sci."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"e02367-17","DOI":"10.1128\/AAC.02367-17","article-title":"The Amphibian Antimicrobial Peptide Temporin B Inhibits In Vitro Herpes Simplex Virus 1 Infection","volume":"62","author":"Marcocci","year":"2018","journal-title":"Antimicrob. Agents Chemother."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"94","DOI":"10.1016\/j.phytochem.2018.07.002","article-title":"Antimicrobial peptides from different plant sources: Isolation, characterisation, and purification","volume":"154","author":"Tang","year":"2018","journal-title":"Phytochemistry"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"329","DOI":"10.1016\/S0145-305X(99)00015-4","article-title":"Antimicrobial peptides in insects; structure and function","volume":"23","author":"Bulet","year":"1999","journal-title":"Dev. Comp. Immunol."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"72","DOI":"10.1016\/j.micpath.2018.04.008","article-title":"Venom-derived peptide Mastoparan-1 eradicates planktonic and biofilm-embedded methicillin-resistant Staphylococcus aureus isolates","volume":"119","author":"Memariani","year":"2018","journal-title":"Microb. Pathog."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"1007","DOI":"10.1016\/j.peptides.2009.01.018","article-title":"Plant defensins\u2014Prospects for the biological functions and biotechnological properties","volume":"30","author":"Gomes","year":"2009","journal-title":"Peptides"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"389","DOI":"10.1038\/415389a","article-title":"Antimicrobial peptides of multicellular organisms","volume":"415","author":"Zasloff","year":"2002","journal-title":"Nature"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"58","DOI":"10.3390\/ph7010058","article-title":"Host-defense peptides with therapeutic potential from skin secretions of frogs from the family pipidae","volume":"7","author":"Conlon","year":"2014","journal-title":"Pharmaceuticals"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"382","DOI":"10.1016\/j.ijantimicag.2003.07.022","article-title":"Antiviral activity of antimicrobial cationic peptides against Junin virus and herpes simplex virus","volume":"23","author":"Castilla","year":"2004","journal-title":"Int. J. Antimicrob. Agents"},{"key":"ref_18","first-page":"3534","article-title":"Antitumor activity of magainin analogues against human lung cancer cell lines","volume":"52","author":"Ohsaki","year":"1992","journal-title":"Cancer Res."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"141","DOI":"10.1016\/j.eururo.2005.12.043","article-title":"Antitumor activity of the antimicrobial peptide magainin II against bladder cancer cell lines","volume":"50","author":"Lehmann","year":"2006","journal-title":"Eur. Urol."},{"key":"ref_20","doi-asserted-by":"crossref","unstructured":"Kim, M.K., Kang, N., Ko, S.J., Park, J., Park, E., Shin, D.W., Kim, S.H., Lee, S.A., Lee, J.I., and Lee, S.H. (2018). Antibacterial and Antibiofilm Activity and Mode of Action of Magainin 2 against Drug-Resistant Acinetobacter baumannii. Int. J. Mol. Sci., 19.","DOI":"10.3390\/ijms19103041"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"11598","DOI":"10.1128\/JVI.79.18.11598-11606.2005","article-title":"Antimicrobial peptides from amphibian skin potently inhibit human immunodeficiency virus infection and transfer of virus from dendritic cells to T cells","volume":"79","author":"VanCompernolle","year":"2005","journal-title":"J. Virol."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"1421","DOI":"10.3389\/fphar.2019.01421","article-title":"Dermaseptins, Multifunctional Antimicrobial Peptides: A Review of Their Pharmacology, Effectivity, Mechanism of Action, and Possible Future Directions","volume":"10","author":"Bartels","year":"2019","journal-title":"Front. Pharmacol."},{"key":"ref_23","doi-asserted-by":"crossref","unstructured":"Stelitano, D., Franci, G., Chianese, A., Galdiero, S., Morelli, G., and Galdiero, M. (2019). HSV membrane glycoproteins, their function in viral entry and their use in vaccine studies. Amino Acids, Peptides and Proteins, Royal Society of Chemistry.","DOI":"10.1039\/9781788013857-00014"},{"key":"ref_24","first-page":"710","article-title":"Atrial natriuretic peptide. New aspects in the regulation of blood volume and electrolyte homeostasis","volume":"134","author":"Tulassay","year":"1986","journal-title":"Monatsschr. Kinderheilkd."},{"key":"ref_25","doi-asserted-by":"crossref","unstructured":"Roy, M., Lebeau, L., Chessa, C., Damour, A., Ladram, A., Oury, B., Boutolleau, D., Bodet, C., and Leveque, N. (2019). Comparison of Anti-Viral Activity of Frog Skin Anti-Microbial Peptides Temporin-Sha and [K(3)]SHa to LL-37 and Temporin-Tb against Herpes Simplex Virus Type 1. Viruses, 11.","DOI":"10.3390\/v11010077"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"496","DOI":"10.1016\/S0006-291X(03)00999-9","article-title":"A melittin-related peptide from the skin of the Japanese frog, Rana tagoi, with antimicrobial and cytolytic properties","volume":"306","author":"Conlon","year":"2003","journal-title":"Biochem. Biophys. Res. Commun."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"317","DOI":"10.1016\/j.ijantimicag.2006.09.007","article-title":"Activities of four frog skin-derived antimicrobial peptides (temporin-1DRa, temporin-1Va and the melittin-related peptides AR-23 and RV-23) against anaerobic bacteria","volume":"29","author":"Urban","year":"2007","journal-title":"Int. J. Antimicrob. Agents"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"27394","DOI":"10.1038\/srep27394","article-title":"Design of an alpha-helical antimicrobial peptide with improved cell-selective and potent anti-biofilm activity","volume":"6","author":"Zhang","year":"2016","journal-title":"Sci. Rep."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"4012394","DOI":"10.1155\/2019\/4012394","article-title":"Melittin Inhibition and Eradication Activity for Resistant Polymicrobial Biofilm Isolated from a Dairy Industry after Disinfection","volume":"2019","author":"Galdiero","year":"2019","journal-title":"Int. J. Microbiol."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"548","DOI":"10.1006\/viro.1993.1510","article-title":"Role of the Na+, K+ pump in herpes simplex type 1-induced cell fusion: Melittin causes specific reversion of syncytial mutants with the syn1 mutation to Syn+ (wild-type) phenotype","volume":"196","author":"Baghian","year":"1993","journal-title":"Virology"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"853","DOI":"10.1007\/s12275-016-6376-1","article-title":"Inhibitory effects of bee venom and its components against viruses in vitro and in vivo","volume":"54","author":"Uddin","year":"2016","journal-title":"J. Microbiol."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"235","DOI":"10.1016\/0014-5793(92)80780-K","article-title":"Influence of amphipathic peptides on the HIV-1 production in persistently infected T lymphoma cells","volume":"309","author":"Wachinger","year":"1992","journal-title":"FEBS Lett."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"6483","DOI":"10.3892\/mmr.2015.4275","article-title":"Melittin, a honeybee venomderived antimicrobial peptide, may target methicillinresistant Staphylococcus aureus","volume":"12","author":"Choi","year":"2015","journal-title":"Mol. Med. Rep."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"95","DOI":"10.1179\/1973947815Y.0000000004","article-title":"Antibacterial and anti-biofilm activities of melittin and colistin, alone and in combination with antibiotics against Gram-negative bacteria","volume":"28","author":"Dosler","year":"2016","journal-title":"J. Chemother."},{"key":"ref_35","doi-asserted-by":"crossref","unstructured":"Falanga, A., Del Genio, V., Kaufman, E.A., Zannella, C., Franci, G., Weck, M., and Galdiero, S. (2021). Engineering of Janus-Like Dendrimers with Peptides Derived from Glycoproteins of Herpes Simplex Virus Type 1: Toward a Versatile and Novel Antiviral Platform. Int. J. Mol. Sci., 22.","DOI":"10.3390\/ijms22126488"},{"key":"ref_36","first-page":"e53124","article-title":"Early Viral Entry Assays for the Identification and Evaluation of Antiviral Compounds","volume":"104","author":"Tai","year":"2015","journal-title":"J. Vis. Exp."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"569967","DOI":"10.3389\/fbioe.2020.569967","article-title":"Combating Actions of Green 2D-Materials on Gram Positive and Negative Bacteria and Enveloped Viruses","volume":"8","author":"Singh","year":"2020","journal-title":"Front. Bioeng. Biotechnol."},{"key":"ref_38","doi-asserted-by":"crossref","unstructured":"Laksono, B.M., de Vries, R.D., McQuaid, S., Duprex, W.P., and de Swart, R.L. (2016). Measles Virus Host Invasion and Pathogenesis. Viruses, 8.","DOI":"10.3390\/v8080210"},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"538","DOI":"10.1055\/s-0036-1584798","article-title":"Parainfluenza Virus Infection","volume":"37","author":"Branche","year":"2016","journal-title":"Semin. Respir. Crit. Care Med."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"1518","DOI":"10.1016\/j.peptides.2011.05.015","article-title":"Virucidal activity of a scorpion venom peptide variant mucroporin-M1 against measles, SARS-CoV and influenza H5N1 viruses","volume":"32","author":"Li","year":"2011","journal-title":"Peptides"},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"e01026-17","DOI":"10.1128\/JVI.01026-17","article-title":"Mutations in the Fusion Protein of Measles Virus That Confer Resistance to the Membrane Fusion Inhibitors Carbobenzoxy-d-Phe-l-Phe-Gly and 4-Nitro-2-Phenylacetyl Amino-Benzamide","volume":"91","author":"Ha","year":"2017","journal-title":"J. Virol."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"241","DOI":"10.1177\/095632020101200405","article-title":"Inhibition of measles virus replication by 5\u2032-nor carbocyclic adenosine analogues","volume":"12","author":"Barnard","year":"2001","journal-title":"Antivir. Chem. Chemother."},{"key":"ref_43","doi-asserted-by":"crossref","unstructured":"Giugliano, R., Buonocore, C., Zannella, C., Chianese, A., Palma Esposito, F., Tedesco, P., De Filippis, A., Galdiero, M., Franci, G., and de Pascale, D. (2021). Antiviral Activity of the Rhamnolipids Mixture from the Antarctic Bacterium Pseudomonas gessardii M15 against Herpes Simplex Viruses and Coronaviruses. Pharmaceutics, 13.","DOI":"10.3390\/pharmaceutics13122121"},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"612","DOI":"10.1111\/j.1440-1681.2006.04415.x","article-title":"Antiviral effects of saikosaponins on human coronavirus 229E in vitro","volume":"33","author":"Cheng","year":"2006","journal-title":"Clin. Exp. Pharmacol. Physiol."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"700502","DOI":"10.3389\/fcimb.2021.700502","article-title":"Combination Treatment with Remdesivir and Ivermectin Exerts Highly Synergistic and Potent Antiviral Activity Against Murine Coronavirus Infection","volume":"11","author":"Tan","year":"2021","journal-title":"Front. Cell Infect. Microbiol."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"633","DOI":"10.1080\/14786419.2019.1582046","article-title":"Anti-poliovirus activity of Nerium oleander aqueous extract","volume":"35","author":"Sanna","year":"2021","journal-title":"Nat. Prod. Res."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"3929","DOI":"10.1128\/JVI.74.8.3929-3931.2000","article-title":"An antiviral compound that blocks structural transitions of poliovirus prevents receptor binding at low temperatures","volume":"74","author":"Dove","year":"2000","journal-title":"J. Virol."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"430","DOI":"10.7150\/jca.29787","article-title":"Identification of Putative UL54 (ICP27) Transcription Regulatory Sequences Binding to Oct-1, v-Myb, Pax-6 and Hairy in Herpes Simplex Viruses","volume":"10","author":"Wang","year":"2019","journal-title":"J. Cancer"},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"1936","DOI":"10.1038\/s41467-021-21953-3","article-title":"The SARS-CoV-2 nucleocapsid protein is dynamic, disordered, and phase separates with RNA","volume":"12","author":"Cubuk","year":"2021","journal-title":"Nat. Commun."},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"28","DOI":"10.1002\/bip.10374","article-title":"Structure-activity relationships of de novo designed cyclic antimicrobial peptides based on gramicidin S","volume":"71","author":"Lee","year":"2003","journal-title":"Biopolymers"},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"1448","DOI":"10.1345\/aph.1D143","article-title":"Enfuvirtide: A novel agent for the treatment of HIV-1 infection","volume":"37","author":"Duffalo","year":"2003","journal-title":"Ann. Pharmacother."},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"259","DOI":"10.1358\/dot.2004.40.3.820089","article-title":"Enfuvirtide","volume":"40","author":"Lalezari","year":"2004","journal-title":"Drugs Today"},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"38","DOI":"10.1016\/j.peptides.2018.02.006","article-title":"Automatic procedures for the synthesis of difficult peptides using oxyma as activating reagent: A comparative study on the use of bases and on different deprotection and agitation conditions","volume":"102","author":"Caporale","year":"2018","journal-title":"Peptides"},{"key":"ref_54","doi-asserted-by":"crossref","first-page":"4888","DOI":"10.1128\/JVI.77.8.4888-4898.2003","article-title":"Membrane association of VP22, a herpes simplex virus type 1 tegument protein","volume":"77","author":"Brignati","year":"2003","journal-title":"J. Virol."},{"key":"ref_55","doi-asserted-by":"crossref","unstructured":"Petrillo, F., Chianese, A., De Bernardo, M., Zannella, C., Galdiero, M., Reibaldi, M., Avitabile, T., Boccia, G., Galdiero, M., and Rosa, N. (2021). Inhibitory Effect of Ophthalmic Solutions against SARS-CoV-2: A Preventive Action to Block the Viral Transmission?. Microorganisms, 9.","DOI":"10.3390\/microorganisms9081550"},{"key":"ref_56","first-page":"50833","article-title":"Production and purification of non replicative canine adenovirus type 2 derived vectors","volume":"82","author":"Szelechowski","year":"2013","journal-title":"J. Vis. Exp."}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/23\/2\/883\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,13]],"date-time":"2025-10-13T14:15:01Z","timestamp":1760364901000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/23\/2\/883"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,1,14]]},"references-count":56,"journal-issue":{"issue":"2","published-online":{"date-parts":[[2022,1]]}},"alternative-id":["ijms23020883"],"URL":"https:\/\/doi.org\/10.3390\/ijms23020883","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2022,1,14]]}}}