Cat�s whiskers (Orthosiphon stamineus) leaves extracts were prepared using supercritical CO2 (SC-CO2) with full factorial design\nto determine the optimum extraction parameters. Nine extracts were obtained by varying pressure, temperature, and time.\nThe extracts were analysed using FTIR, UV-Vis, and GC-MS. Cytotoxicity of the extracts was evaluated on human (colorectal,\nbreast, and prostate) cancer and normal fibroblast cells. Moderate pressure (31.1MPa) and temperature (60?C) were recorded\nas optimum extraction conditions with high yield (1.74%) of the extract (B2) at 60 min extraction time. The optimized extract\n(B2) displayed selective cytotoxicity against prostate cancer (PC3) cells (IC50 28 ????g/mL) and significant antioxidant activity (IC50\n42.8 ????g/mL). Elevated levels of caspases 3/7 and 9 in B2-treated PC3 cells suggest the induction of apoptosis through nuclear and\nmitochondrial pathways. Hoechst and rhodamine assays confirmed the nuclear condensation and disruption of mitochondrial\nmembrane potential in the cells. B2 also demonstrated inhibitory effects on motility and colonies of PC3 cells at its subcytotoxic\nconcentrations. It is noteworthy that B2 displayed negligible toxicity against the normal cells. Chemometric analysis revealed high\ncontent of essential oils, hydrocarbon, fatty acids, esters, and aromatic sesquiterpenes in B2. This study highlights the therapeutic\npotentials of SC-CO2 extract of cat�s whiskers in targeting prostate carcinoma.
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