Urokinase plasminogen activator inhibitory and anti-breast cancer activity of Cananga odorata and Lantana camara leaf extracts from optimization of ultrasound-assisted extraction

Background: Cancer invasion and metastasis are complex processes that depend on the degradation of the extracellular matrix, largely facilitated by proteolytic enzymes such as urokinase-type plasminogen activator (uPA). Elevated levels of uPA have been consistently correlated with increased tumor aggressiveness and poorer clinical outcomes in breast cancer patients, making this enzyme a key therapeutic target. Flavonoids found in the hydroalcoholic leaf extracts of Cananga odorata (CO) and Lantana camara (LC) have previously been associated with inhibitory effects on cancer invasion and metastasis. Objectives: This study aimed to investigate the influence of different Ultrasound Assisted Extraction (UAE) method parameters, including ethanol concentration (50–90%), extraction time (10–50 minute), and temperature extraction (30–60 0C). The optimal UAE condition from CO and LC on total flavonoid content (TFC) and antioxidant activity (AA) were determined using Response Surface Methodology (RSM). The biological activities of the optimized extracts were further evaluated as uPA inhibition and cytotoxicity test. Methods: The TFC and AA were quantified using AlCl3 and DPPH assays, respectively. Additionally, the Urokinase Inhibitor Screening Kit was utilized to assess uPA inhibition, while the PrestoBlue assay was employed to measure cytotoxicity test against MCF-7 human breast cancer cells. Results: The optimal UAE conditions were 74% ethanol concentration, 31 minutes of extraction time, and an extraction temperature of 41°C resulted in extracts with high bioactive content, while LC showing superior TFC and stronger AA compared to CO. The results demonstrated that LC exhibited more potent bioactivities than CO leaf extract, with significantly lower IC50 values for uPA inhibition and cytotoxicity test were 24.13 ± 2.68 ?g/mL and 42.87 ± 2.24 ?g/mL). Conclusion: These findings suggest that LC extract effectively suppresses uPA activity while exerting cytotoxic effects on breast cancer cells, indicating its potential as a anticancer and chemoprevention agent.