In vitro antiviral activities of thymol and Limonin against influenza a viruses and SARS-CoV-2

Abstract Emerging and re-emerging respiratory viruses represent a continuing threat to human health. The pandemic severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) and influenza A viruses (IAVs) are co-circulating, presenting serious threats to public health. Therefore, screening for safe...

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Main Authors: Gomaa Mostafa-Hedeab, Akram Hegazy, Islam Mostafa, Ibrahim H. Eissa, Ahmed M. Metwaly, Hany Abdelfattah Elhady, Abozer Y. Eledrdery, Sager Holyl Alruwaili, Alaa Oqalaa E. Alibrahim, Fawaz O. Alenazy, Muhannad Faleh Alanazi Alruwaili, Thamer Alshami Marghel, Muharib Alruwaili, Alaaeldin Mohamed Saad, Mahmoud Bayoumi, Assem Mohamed El-Shazly, Luis Martinez-Sobrido, Ahmed Mostafa
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Language:English
Published: Nature Portfolio 2025-07-01
Series:Scientific Reports
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Online Access:https://doi.org/10.1038/s41598-025-06967-x
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author Gomaa Mostafa-Hedeab
Akram Hegazy
Islam Mostafa
Ibrahim H. Eissa
Ahmed M. Metwaly
Hany Abdelfattah Elhady
Abozer Y. Eledrdery
Sager Holyl Alruwaili
Alaa Oqalaa E. Alibrahim
Fawaz O. Alenazy
Muhannad Faleh Alanazi Alruwaili
Thamer Alshami Marghel
Muharib Alruwaili
Alaaeldin Mohamed Saad
Mahmoud Bayoumi
Assem Mohamed El-Shazly
Luis Martinez-Sobrido
Ahmed Mostafa
author_facet Gomaa Mostafa-Hedeab
Akram Hegazy
Islam Mostafa
Ibrahim H. Eissa
Ahmed M. Metwaly
Hany Abdelfattah Elhady
Abozer Y. Eledrdery
Sager Holyl Alruwaili
Alaa Oqalaa E. Alibrahim
Fawaz O. Alenazy
Muhannad Faleh Alanazi Alruwaili
Thamer Alshami Marghel
Muharib Alruwaili
Alaaeldin Mohamed Saad
Mahmoud Bayoumi
Assem Mohamed El-Shazly
Luis Martinez-Sobrido
Ahmed Mostafa
author_sort Gomaa Mostafa-Hedeab
collection DOAJ
description Abstract Emerging and re-emerging respiratory viruses represent a continuing threat to human health. The pandemic severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) and influenza A viruses (IAVs) are co-circulating, presenting serious threats to public health. Therefore, screening for safe and broad-spectrum antiviral candidates to control such viral infections is prioritized. Herein, this study reports the in vitro antiviral activity of some essential volatile oils (EOs) and volatile oil components including Peppermint oil, Eucalyptus oil, Clove oil, Thymol, Camphor and Limonin against two different IAVs, namely influenza A/H1N1 and A/H5N1 viruses, and SARS-CoV-2 virus. All tested samples were safe in MDCK and Vero E6 cell lines with CC50 values that exceed 1 mg/ml, allowing the screening of their antiviral activities using a wide range of concentrations. The results show the potency of Thymol and Limonin against influenza A/H1N1 virus with IC50 values of 0.022 and 4.25 µg/ml, respectively. The anti-influenza activities of Thymol and Limonin were further validated by testing them against the avian influenza A/H5N1 virus, resulting in anti-influenza activities with IC50 values of 18.5 and 15.6 ng/ml, respectively. The broad-spectrum potential of the highly potent antiviral candidates, Thymol and Limonin, were further tested against the pandemic SARS-CoV-2 and, both exerted anti-coronavirus activities with IC50 values of 0.591 and 4.04 µg/ml, respectively. Further investigations against influenza A/H1N1 virus revealed that Thymol and Limonin could inhibit IAV by hindering viral replication. The Biochemical analyses of the interaction of Limonin and Thymol with FDA-approved anti-influenza drug targets, neuraminidase and viral polymerases, revealed that both compounds can partially inhibit IAV polymerase activity, but have no effect on neuraminidase activity. Likely, molecular docking studies indicated that Thymol and Limonin obstruct active binding sites of IAV polymerases. These findings presented on the antiviral activity of Limonin and Thymol might be used to support the development of supplemental therapy against currently emerging and reemerging respiratory viral infections.
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spelling doaj-art-a4bbb3a2881b45b48f00bc17f92ce5c02025-08-20T03:45:28ZengNature PortfolioScientific Reports2045-23222025-07-0115111510.1038/s41598-025-06967-xIn vitro antiviral activities of thymol and Limonin against influenza a viruses and SARS-CoV-2Gomaa Mostafa-Hedeab0Akram Hegazy1Islam Mostafa2Ibrahim H. Eissa3Ahmed M. Metwaly4Hany Abdelfattah Elhady5Abozer Y. Eledrdery6Sager Holyl Alruwaili7Alaa Oqalaa E. Alibrahim8Fawaz O. Alenazy9Muhannad Faleh Alanazi Alruwaili10Thamer Alshami Marghel11Muharib Alruwaili12Alaaeldin Mohamed Saad13Mahmoud Bayoumi14Assem Mohamed El-Shazly15Luis Martinez-Sobrido16Ahmed Mostafa17Pharmacology Department and Health Research Unit, Medical College, Jouf UniversityDepartment of Agricultural Microbiology, Faculty of Agriculture, Cairo UniversityDepartment of Pharmacognosy, Faculty of Pharmacy, Zagazig UniversityMedicinal Chemistry Department, Faculty of Pharmacy (Boys), Al-Azhar UniversityPharmacognosy and Medicinal Plants Department, Faculty of Pharmacy (Boys), Al- Azhar UniversitySurgery Department, Medical College, Jouf UniversityDepartment of Clinical Laboratory Sciences, College of Applied Medical Sciences, Jouf UniversityDepartment of Surgery, Orthopedic Division, College of Medicine, Jouf UniversityDepartment of Internal Medicine, College of Medicine, Jouf UniversityDepartment of Clinical Laboratory Sciences, College of Applied Medical Sciences, Jouf UniversityDepartment of Internal Medicine, Division of Radiology, College of Medicine, Jouf University, Kingdom of Saudi ArabiaDepartment of Pediatrics, College of Medicine, Jouf University, Jouf University, Kingdom of Saudi ArabiaDepartment of Clinical Laboratory Sciences, College of Applied Medical Sciences, Jouf UniversityDepartment of Zoonoses, Faculty of Veterinary Medicine, Zagazig UniversityDisease Intervention & Prevention (DIP) and Host Pathogen Interactions (HPI), Texas Biomedical Research InstituteDepartment of Pharmacognosy, Faculty of Pharmacy, Zagazig UniversityDisease Intervention & Prevention (DIP) and Host Pathogen Interactions (HPI), Texas Biomedical Research InstituteDisease Intervention & Prevention (DIP) and Host Pathogen Interactions (HPI), Texas Biomedical Research InstituteAbstract Emerging and re-emerging respiratory viruses represent a continuing threat to human health. The pandemic severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) and influenza A viruses (IAVs) are co-circulating, presenting serious threats to public health. Therefore, screening for safe and broad-spectrum antiviral candidates to control such viral infections is prioritized. Herein, this study reports the in vitro antiviral activity of some essential volatile oils (EOs) and volatile oil components including Peppermint oil, Eucalyptus oil, Clove oil, Thymol, Camphor and Limonin against two different IAVs, namely influenza A/H1N1 and A/H5N1 viruses, and SARS-CoV-2 virus. All tested samples were safe in MDCK and Vero E6 cell lines with CC50 values that exceed 1 mg/ml, allowing the screening of their antiviral activities using a wide range of concentrations. The results show the potency of Thymol and Limonin against influenza A/H1N1 virus with IC50 values of 0.022 and 4.25 µg/ml, respectively. The anti-influenza activities of Thymol and Limonin were further validated by testing them against the avian influenza A/H5N1 virus, resulting in anti-influenza activities with IC50 values of 18.5 and 15.6 ng/ml, respectively. The broad-spectrum potential of the highly potent antiviral candidates, Thymol and Limonin, were further tested against the pandemic SARS-CoV-2 and, both exerted anti-coronavirus activities with IC50 values of 0.591 and 4.04 µg/ml, respectively. Further investigations against influenza A/H1N1 virus revealed that Thymol and Limonin could inhibit IAV by hindering viral replication. The Biochemical analyses of the interaction of Limonin and Thymol with FDA-approved anti-influenza drug targets, neuraminidase and viral polymerases, revealed that both compounds can partially inhibit IAV polymerase activity, but have no effect on neuraminidase activity. Likely, molecular docking studies indicated that Thymol and Limonin obstruct active binding sites of IAV polymerases. These findings presented on the antiviral activity of Limonin and Thymol might be used to support the development of supplemental therapy against currently emerging and reemerging respiratory viral infections.https://doi.org/10.1038/s41598-025-06967-xRespiratory virusesSARS-CoV-2IAVEssential oilsLimoninThymol
spellingShingle Gomaa Mostafa-Hedeab
Akram Hegazy
Islam Mostafa
Ibrahim H. Eissa
Ahmed M. Metwaly
Hany Abdelfattah Elhady
Abozer Y. Eledrdery
Sager Holyl Alruwaili
Alaa Oqalaa E. Alibrahim
Fawaz O. Alenazy
Muhannad Faleh Alanazi Alruwaili
Thamer Alshami Marghel
Muharib Alruwaili
Alaaeldin Mohamed Saad
Mahmoud Bayoumi
Assem Mohamed El-Shazly
Luis Martinez-Sobrido
Ahmed Mostafa
In vitro antiviral activities of thymol and Limonin against influenza a viruses and SARS-CoV-2
Scientific Reports
Respiratory viruses
SARS-CoV-2
IAV
Essential oils
Limonin
Thymol
title In vitro antiviral activities of thymol and Limonin against influenza a viruses and SARS-CoV-2
title_full In vitro antiviral activities of thymol and Limonin against influenza a viruses and SARS-CoV-2
title_fullStr In vitro antiviral activities of thymol and Limonin against influenza a viruses and SARS-CoV-2
title_full_unstemmed In vitro antiviral activities of thymol and Limonin against influenza a viruses and SARS-CoV-2
title_short In vitro antiviral activities of thymol and Limonin against influenza a viruses and SARS-CoV-2
title_sort in vitro antiviral activities of thymol and limonin against influenza a viruses and sars cov 2
topic Respiratory viruses
SARS-CoV-2
IAV
Essential oils
Limonin
Thymol
url https://doi.org/10.1038/s41598-025-06967-x
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