Plants are important natural resources used for many purposes. They especially stand out with their medical potential. In this study, the extract conditions showing the highest biological activity were optimized using the artificial intelligence application of Salvia cilicica Boiss. Then, the biological activities of the optimized extract were determined. Response Surface Method was used for optimization. Using the Box-Behnken experimental design, antioxidant capacities of the optimized extracts were evaluated through Rel Assay TAS and TOS kits, as well as DPPH and FRAP methods. Anticholinesterase activity was assessed by measuring acetyl- and butyrylcholinesterase enzyme activities. Total phenolic content was quantified using the Folin-Ciocalteu reagent, while total flavonoid content was analyzed via the aluminum chloride assay. The antiproliferative potential was examined against the A549 lung cancer cell line, and phenolic compounds were identified using an LC-MS/MS system. Optimization results indicated that the ideal extraction parameters were 45.890 °C, 7.730 h, and a 96.431 ethanol-to-water ratio. The extract obtained under these conditions exhibited a TAS value of 7.461 ± 0.065 mmol/L, a TOS value of 5.630 ± 0.163 µmol/L, and an OSI value of 0.075 ± 0.002. Total phenolic and flavonoid contents were measured as 97.681 ± 1.076 mg/g and 113.067 ± 0.621 mg/g, respectively. The FRAP value reached 70.669 ± 0.199 mg/g, while the DPPH value was determined as 53.208 ± 0.427 mg/g. Furthermore, anti-AChE and anti-BChE activities were recorded at 12.93 ± 0.72 µg/mL and 25.87 ± 1.44 µg/mL, respectively. The extract demonstrated notable cytotoxicity against the A549 cancer cell line. Additionally, 10 distinct phenolic compounds were identified in the plant. These findings highlight the significant biological potential of S. cilicica extract obtained under optimal conditions, suggesting its potential as a valuable natural resource in pharmacological applications.