{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,4,1]],"date-time":"2026-04-01T11:02:12Z","timestamp":1775041332269,"version":"3.50.1"},"reference-count":43,"publisher":"MDPI AG","issue":"3","license":[{"start":{"date-parts":[[2020,3,11]],"date-time":"2020-03-11T00:00:00Z","timestamp":1583884800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100012245","name":"Science and Technology Planning Project of Guangdong Province","doi-asserted-by":"publisher","award":["2014A020210018"],"award-info":[{"award-number":["2014A020210018"]}],"id":[{"id":"10.13039\/501100012245","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100012245","name":"Science and Technology Planning Project of Guangdong Province","doi-asserted-by":"publisher","award":["2016A020217014"],"award-info":[{"award-number":["2016A020217014"]}],"id":[{"id":"10.13039\/501100012245","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100010256","name":"Guangzhou Municipal Science and Technology Project","doi-asserted-by":"publisher","award":["201604020171"],"award-info":[{"award-number":["201604020171"]}],"id":[{"id":"10.13039\/501100010256","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Viruses"],"abstract":"<jats:p>The objective of this study was to investigate the inhibitory effect of chlorogenin 3-O-\u03b2-chacotrioside derivatives against H5N1 subtype of the highly pathogenic avian influenza (HPAI) viruses and its molecular mechanism. A series of novel small molecule pentacyclic triterpene derivatives were designed and synthesized and their antiviral activities on HPAI H5N1 viruses were detected. The results displayed that the derivatives UA-Nu-ph-5, XC-27-1 and XC-27-2 strongly inhibited wild-type A\/Duck\/Guangdong\/212\/2004 H5N1 viruses with the IC50 values of 15.59 \u00b1 2.4 \u03bcM, 16.83 \u00b1 1.45 \u03bcM, and 12.45 \u00b1 2.27 \u03bcM, respectively, and had the selectivity index (SI) &gt; 3, which was consistent with the efficacy against A\/Thailand\/kan353\/2004 pseudo-typed viruses. Four dealt patterns were compared via PRNT. The prevention dealt pattern showed the strongest inhibitory effects than other patterns, suggesting that these derivatives act on the entry process at the early stages of H5N1 viral infection, providing protection for cells against infection. Further studies through hemagglutinin inhibition (HI) and neuraminidase inhibitory (NAI) assay confirmed that these derivatives inhibited H5N1 virus replication by interfering with the viral hemagglutinin function. The derivatives could recognize specifically HA protein with binding affinity constant KD values of 2.57 \u00d7 10\u22124 M and 3.67 \u00d7 10\u22124 M. In addition, through site-directed mutagenesis combined with a pseudovirion system, we identified that the high-affinity docking sites underlying interaction were closely associated with amino acid residues I391 and T395 of HA. However, the potential binding sites of the derivatives with HA did not locate at HA1 sialic acids receptor binding domain (RBD). Taken together, these study data manifested that chlorogenin 3-O-\u03b2-chacotrioside derivatives generated antiviral effect against HPAI H5N1 viruses by targeting the hemagglutinin fusion machinery.<\/jats:p>","DOI":"10.3390\/v12030304","type":"journal-article","created":{"date-parts":[[2020,3,12]],"date-time":"2020-03-12T04:13:57Z","timestamp":1583986437000},"page":"304","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":7,"title":["Study on the Antiviral Activities and Hemagglutinin-Based Molecular Mechanism of Novel Chlorogenin 3-O-\u03b2-Chacotrioside Derivatives against H5N1 Subtype Viruses"],"prefix":"10.3390","volume":"12","author":[{"given":"Wan-Zhen","family":"Shi","sequence":"first","affiliation":[{"name":"Department of Pharmaceutical Engineering, South China Agricultural University, Guangzhou 510640, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ling-Zhi","family":"Jiang","sequence":"additional","affiliation":[{"name":"Shenzhen Key Laboratory of Marine Bioresource and Eco-Environmental Science, College of Life Sciences and Oceanography, Shenzhen University, Shenzhen 518060, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Gao-Peng","family":"Song","sequence":"additional","affiliation":[{"name":"Department of Pharmaceutical Engineering, South China Agricultural University, Guangzhou 510640, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Sheng","family":"Wang","sequence":"additional","affiliation":[{"name":"Key Laboratory of Molecular Biophysics of the Ministry of Education, College of Life Science and Technology, Huazhong University of Science and Technology, Wuhan 430074, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ping","family":"Xiong","sequence":"additional","affiliation":[{"name":"Department of Pharmaceutical Engineering, South China Agricultural University, Guangzhou 510640, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Chang-Wen","family":"Ke","sequence":"additional","affiliation":[{"name":"Guangdong Provincial Center for Disease Control and Prevention, Guangzhou 511430, China"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2020,3,11]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"15","DOI":"10.1006\/viro.1999.9820","article-title":"Genetic characterization of the pathogenic influenzaA\/Goose\/Guangdong\/1\/96 (H5N1) virus: Similarity of its hemagglutinin gene to those of H5N1 viruses from the 1997 outbreaks in Hong Kong","volume":"261","author":"Xu","year":"1999","journal-title":"Virology"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"7529","DOI":"10.1128\/JVI.00327-07","article-title":"Characterization of low-pathogenic H5 subtype influenza viruses from Eurasia: Implications for the origin of highly pathogenic H5N1 viruses","volume":"81","author":"Duan","year":"2007","journal-title":"J. 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