Currently, ovarian cancer exhibits a high mortality rate. Eucommia, a traditional Chinese medicine and food, has been demonstrated anti-tumor effects, potentially providing effective treatments for ovarian cancer. The inhibitory effects of Eucommia Folium extract (EFe) on ovarian cancer cells were examined by a series of in vitro assays. The key targets and related metabolites were identified by network pharmacology and metabolomics. Furthermore, the molecules in EFe were bound to targets via molecular docking. EFe had significant inhibitory effects on the proliferation, adhesion and migration of ovarian cancer cells. At a concentration of 0.3 mg/mL, EFe inhibited ovarian cancer cells by about 15%. It also showed migration inhibition rates of 55% on OVCAR-3 and 30% on SK-OV-3. Additionally, EFe demonstrated adhesion inhibition rates of 28% on OVCAR-3 and 20% on SK-OV-3. Metabolomics analysis identified 33 differential metabolites, primarily related to fatty acid biosynthesis and degradation, and tyrosine metabolism, with 17 being down-regulated and 16 up-regulated. The network pharmacology revealed that kaempferol, (+)-catechin, and quercetin were the active components of EFe, with 58 potential targets identified in EFe against ovarian cancer. The network pharmacology and metabolomics analysis revealed that EFe mainly affected tyrosine metabolism and arachidonatic acid metabolism. The results of molecular docking and molecular dynamics simulation showed that the active molecules in EFe bound strongly to prostaglandin-endoperoxide synthase 2 (PTGS2) and cytochrome P450 family 1 subfamily B member 1 (CYP1B1). The current study showed that EFe had anti-ovarian cancer effects by regulating fatty acid and tyrosine metabolism, possibly through PTGS2 and CYP1B1.