Abstract <p>In an attempt to develop new scaffolds with anticancer potential, a series of new (<i>E</i>)-3-(substitutedbenzylidene)-1,8-naphthyridine-2,4(1<i>H</i>,3<i>H</i>)-dione derivatives was synthesized. The structures of the synthesized compounds were confirmed utilizing different spectroscopic techniques (FT-IR, ¹H, ¹³C NMR, and MS). Investigation of the anti-cervical cancer activity with IC₅₀ values ranging from 320 to 170 μM of all newly synthesized compounds, showed promising activity of all the derivatives against HeLa cell line. Out of the 8 synthesized derivatives, (<i>E</i>)-3-(3-methylbenzylidene)-1,8-naphthyridine-2,4(1<i>H</i>,3<i>H</i>)-dione exhibited the most promising inhibitory activity. Some notations about structure-activity relationship (SAR) offered insight that the presence of electron withdrawing groups (EWG) augmented the activity while electron donating groups (EDG) decreased the activity. Docking studies further supported the findings, and provided the basis of linkages in terms of conventional hydrogen bonds and π-bonding patterns.</p>