Miftahul Mushlih, Marlita Marlita, Kavana H. Kusuma, Yuyun I. Christina, Dinia R. Dwijayanti, Dewi Mustikaningtyas, Nashi Widodo, Muhammad S. Djati, Sri Widyarti, Muhaimin Rifa’i
Global public health systems are facing challenges due to the COVID-19 pandemic, which is caused by the Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2). Given increasing interest in plant-based antivirals, this study determined the potential of ethanol extract of red ginger (Zingiber officinale) to inhibit the attachment of SARS-CoV-2 virus-like particles (VLPs) to human bronchial epithelial cells. SARS-CoV-2 VLPs carrying the D614G spike mutation were generated in HEK293T cells using plasmids encoding the envelope (E), membrane (M), and spike-EGFP fusion proteins. EGFP was used to visualize and quantify VLP attachment. VLP formation was confirmed by fluorescence microscopy and transmission electron microscopy. Cells were exposed to 200 ng/mL of VLPs together with Z. officinale extract at 0.67 to 20 µg/mL for 24-48 h, and viral attachment was quantified via EGFP fluorescence intensity. For in silico analysis, identified compounds of Z. officinale extract were screened based on drug-likeness parameters for further processing to molecular docking and molecular dynamics simulations. The results showed that treatment with Z. officinale extract significantly reduced VLP attachment at concentrations of 0.67–10 µg/mL after 24 h. After 48 h, the lowest concentrations (0.67 and 1.25 µg/mL) maintained inhibitory effects from SARS-CoV-2 VLP, as indicated by reduced EGFP fluorescence. Of the 22 initially screened compounds, 9 met drug-likeness parameters. Among these, (2E)-3-(3,4-dimethoxyphenyl)prop-2-enoic acid and ferulic acid exhibited the strongest binding affinities to the SARS-CoV-2 spike protein (-5.3 and-5.5 kcal/mol, respectively). These findings suggest that Z. officinale contains bioactive compounds with potential as antiviral agents targeting SARS-CoV-2 variants. © 2025 Mushlih et al.
Doctoral Program, Department of Biology, Faculty of Mathematics and Natural Sciences, Brawijaya University, East Java, Malang, Indonesia; Faculty of Health Science, Universitas Muhammadiyah Sidoarjo, East Java, Sidoarjo, Indonesia; Faculty of Mechatronics, Informatics and Interdisciplinary Studies, Technical University of Liberec, Liberec, Czech Republic; Institute for Nanomaterials, Advanced Technologies and Innovation, Technical University of Liberec, Liberec, Czech Republic; Dewan Jamu Indonesia East Java Region, East Java, Malang, Indonesia; Innovation Center of Integrative Jamu and Eco-pharmaca, Brawijaya University, East Java, Malang, 65145, Indonesia; Department of Biology, Faculty of Mathematics and Natural Sciences, Brawijaya University, East Java, Malang, Indonesia; Department of Biology, Faculty of Mathematics and Natural Sciences, Universitas Negeri Semarang, Semarang, Indonesia
