Pharmacological Profiling of <i>Calotropis Procera</i> and <i>Rhazya Stricta</i>: Unraveling the Antibacterial and Anti-Cancer Potential of Chemically Active Metabolites

<b>Background:</b> The increasing prevalence of cancer and bacterial resistance necessitates more effective anti-cancer and anti-bacterial treatments. This study explores the potential of medicinal plants, specifically <i>Calotropis procera</i> (<i>C. procera</i>) and <i>Rhazya stricta</i> (<i>R. stricta</i>), in addressing this need, aiming to uncover new therapeutic interventions. <b>Methods:</b> Various extraction methods for the leaves of <i>C. procera</i> and <i>R. stricta</i> were employed to investigate the anti-bacterial and anti-cancer properties of these herbs. For anti-bacterial testing, extracts were prepared using water, chloroform, and ethyl acetate, and their activity against methicillin-resistant <i>Staphylococcus aureus</i> (<i>S. aureus</i>) (MRSA) and <i>Escherichia coli</i> (<i>E. coli</i>) was assessed. The anti-cancer potential was evaluated through MTT cytotoxicity assays on various cancer cell lines and further testing using high-content imaging (HCI)-Apoptosis Assay and the ApoTox-GloTM Triplex Assay. Liquid chromatography-mass spectrometry (LC-MS) was used to identify the secondary metabolites of <i>C. procera</i>, and computational predictions were performed to assess the activity of these metabolites. <b>Results:</b> The leaf extracts of both <i>C. procera</i> and <i>R. stricta</i> demonstrated antibacterial activity against <i>S. aureus</i> and <i>E. coli</i>. The <i>C. procera</i> ethyl acetate extract exhibited potent anti-cancer effects on several cancer cell lines. The research also revealed a dose-dependent induction of apoptosis and a decline in cell viability. Computational predictions suggested the identified metabolites were active as nuclear receptor ligands and enzyme inhibitors, with good oral bioavailability. Most metabolites were found to be immunologic and cytotoxic, except for proceragenin and calotropone, which were determined to be non-cardiotoxic. <b>Conclusion:</b> The study's findings demonstrate the remarkable cytotoxic and antibacterial effects of <i>C. procera</i> extracts prepared using ethyl acetate. These results pave the way for further <i>in vitro</i> studies to explore the full potential of these extracts and highlight the presence of chemically active metabolites in <i>C. procera</i>, which hold promise as lead molecules for the development of novel therapies targeting bacterial infections and cancer while minimizing potential side effects.