<i>In Vitro</i> and <i>In Silico</i> Investigations of <i>Lepidium sativum</i> Compounds as Promising Antibacterial Agents Against MRSA

<i>Staphylococcus aureus</i> is one of the most prevalent bacterial pathogens of humans, and the treatment of methicillin-resistant <i>Staphylococcus aureus</i> (MRSA) infections poses significant health care challenges worldwide. MRSA often harbors a gene encoding for a specialized transpeptidase enzyme called penicillin-binding protein 2a (PBP2a), which reduces susceptibility to β-lactam antibiotics and confers resistance. To address this issue, our research aimed to identify potential natural drug candidates derived from the leaves of <i>Lepidium sativum L</i>. (<i>L. sativum</i>) using the marker protein PBP2a. Through gas chromatography/mass spectrometry analysis, we isolated five phytochemical compounds from <i>L. sativum</i> and subjected them to molecular docking analysis against the target bacterial protein. Two compounds (PubChem CID: 39186 and 888955) were selected for further evaluation based on their favorable docking scores (-7.1 and -6.3 kcal/mol, respectively). This computational investigation underscored the potential of these compounds as promising antibacterial candidates against the MRSA PBP2a protein. <i>In vitro</i> antibacterial assays using <i>L. sativum</i> extracts demonstrated a minimum inhibitory concentration of 15 μg/mL. Well-diffusion tests exhibited inhibition zones ranging from 12.3 ± 0.5 mm to 19 ± 0.5 mm at different concentrations, while time-kill kinetics over 24 h indicated effective bacterial growth inhibition. Statistical analysis (one-way analysis of variance) confirmed significant concentration-dependent antibacterial activity (<i>p</i> < 0.01). To validate the efficacy of these compounds against MRSA, further <i>in vivo</i> studies are strongly recommended.