Three new series of 4-substituted-5-alkylidene-2,5-dihydrofuran-2-ones were synthesized. The in vitro activity test results showed that some of them exhibited good antibacterial and cytotoxic activities. Among them compound5cshowed the most potent antibacterial activity againstEscherichia coliwith the MIC value of 20.00 μg/mL. Compound9cshowed good cytotoxic activity against Ec9706 cells withIC50value of 19.39 μM, better than that of the reference compound fluorouracil (IC50=37.74 μM).