Introduction: Plant products are a rich source of pharmacologically active molecules and are considered an important and attractive field of scientific investigation for the development of new drugs. The present study aimed to assess the α-amylase, α-glucosidase and lipase enzymes inhibitory activities and the cytotoxic effects of Carlina curetum four solvents fractions in order to better understand the anti-obesity, hypoglycemic and anticancer effects of this plant. Methods: Several qualitative and quantitative phytochemical tests were performed on Carlina curetum solvent fractions utilizing standard phytochemical procedures, followed by an investigation into their ability to inhibit the enzymes α-amylase, α-glucosidase, and lipase and an assessment of cytotoxic activity against HeLa and Colo205 cells using standard biochemical and biotechnological methods. Results: The results revealed that the aqueous and methanol fractions had the highest α-amylase enzyme inhibitory activity with IC50 values of 21.37 ± 0.31 and 30.2 ± 0.42 μg/mL, respectively, in comparison with Acarbose, which had an IC50 value of 28.18 ± 0.42 μg/mL. The methanol fraction showed potent α-glucosidase inhibitory activity with an IC50 value of 27.54 ± 4.28 μg/mL; the α-glucosidase inhibitory activity Acarbose was 37.15 ± 0.33 μg/mL. The hexane fraction had greater anti-lipase activity than Orlistat. In addition, 0.5 mg/ mL of the C. curetum acetone and hexane fractions had pronounced cytotoxic effects on the Colo-205 cancer cell line, and 0.625 mg/ml of the C. curetum hexane fraction had potential cytotoxic effects against the cervical epithelial carcinoma (HeLa) cell line. Conclusion: This study revealed that C. curetum has potential α-amylase, α-glucosidase, porcine pancreatic lipase enzyme inhibitory activity and cytotoxic activity against the HeLa and Colo-205 cancer cell lines, which indicates the presence of biologically active and cytotoxic compounds in this plant species. This may be considered a challenge for developing bioactive compounds in diabetes, obesity and cancer management.