Cousinia is one of the widespread genera of Asteraceae family. This study aims to evaluate the biological activities and phytochemical compositions of C. iconica extracts. The antioxidant effects of the extracts were investigated on DPPH•, ABTS+•, and iron chelating methods, and the inhibition effects were tested on α-amylase, α-glucosidase, and tyrosinase, as well as AChE, and BChE enzymes. The quantitative and qualitative determinations of the compounds of the extracts were carried out by LC-MS/MS. As a result, the highest TPC (271.65 ± 3.32 mg GAE/gextract), and TFC (4.79 ± 0.06 mg QE/gextract) were found in C. iconica ethyl acetate (CIE) sub-extract. The highest antioxidant activity on DPPH • (IC50 = 90.397 ± 0.575 µg/mL) was shown in C. iconica methanol (CIM) extract. The highest ABTS +•, and iron chelating activities were found in CIE with IC50 = 70.70 ± 0.38, and 615.10 ± 2.47 µg/mL, respectively. The highest α-amylase, and tyrosinase activities were found in CIE, and C. iconica n-butanol sub-extract (CIB) with 864.57 ± 0.68, and 475.21 ± 6.22 µg/mL IC50 values, respectively. AChE, and BChE activities were observed as the highest in CIB (IC50 = 426.50 ± 3.78 µg/mL), and CIM (IC50 = 330.54 ± 0.79 µg/mL), respectively. According to LC-MS/MS analyses, rutin, and cynarin were determined as the main constituents of CIM, and CIE. This is the first report that these biological activities and phytochemical compositions of C. iconica were investigated.