Walid Bakeer ¹ Marwa Gaafar ¹ Ahmed O. El-Gendy ¹ M A. El Badry ² Mona G.Khalil ³ Abdallah T. Mansour ⁴ Nada Alharbi ⁵ Heba Selim ⁶ Mahmoud M. Bendary ^7*

• ¹ Beni-Suef University, Beni-Suef, Beni-Suef, Egypt
• ² Faculty of Science, Al Azhar University, Cairo, Beni Suef, Egypt
• ³ Modern University for Information and Technology, Cairo, Cairo, Egypt
• ⁴ Alexandria University, Alexandria, Alexandria, Egypt
• ⁵ Princess Nourah bint Abdulrahman University, Riyadh, Saudi Arabia
• ⁶ College of Pharmacy, Al Maaref University College, Ramadi, Al Anbar, Iraq
• ⁷ Port Said University, Port Said, Egypt

Despite years of efforts to develop new antibiotics for eradicating multidrug-resistant (MDR) and multi-virulent Methicillin-Resistant Staphylococcus aureus (MRSA) and Vancomycin-Resistant Staphylococcus aureus (VRSA) infections, treatment failures and poor prognoses in most cases have been common. Therefore, there is an urgent need for new therapeutic approaches targeting virulence arrays. Our aim is to discover new anti-virulence therapies targeting MRSA and VRSA virulence arrays. We employed phenotypic, molecular docking, and genetic studies to screen for anti-virulence activities among selected promising compounds: Coumarin, Simvastatin, and Ibuprofen. We found that nearly all detected MRSA and VRSA strains exhibited MDR and multi-virulent profiles. The molecular docking results aligned with the phenotypic and genetic assessments of virulence production. Biofilm and hemolysin productions were inhibited, and all virulence genes were downregulated upon treatment with sub-minimum inhibitory concentration (sub-MIC) of these promising compounds.Ibuprofen was the most active compound, exhibiting the highest inhibition and downregulation of virulence gene products. Moreover, in vivo and histopathological studies confirmed these results. Interestingly, we observed a significant decrease in wound area and improvements in re-epithelialization and tissue organization in the Ibuprofen and antimicrobial treated group compared to the antimicrobial treated group alone. These findings support the idea that a combination of Ibuprofen and antimicrobial drugs may offer a promising new therapy for MRSA and VRSA infections. We hope that our findings can be implemented in clinical practice to assist physicians in making the most suitable treatment decisions.