Design and Multi-Level Biological Evaluation of Naphthyridine-Based Derivatives as Topoisomerase I/II-Targeted Anticancer Agents with Anti-Fowlpox Virus Activity Supported by In Silico Analysis

Abstract

Naphthyridine derivatives have emerged as privileged scaffolds with diverse pharmacological activities, particularly in anticancer and antiviral drug discovery. In this study, a series of naphthyridine-based derivatives (<b>1</b>-<b>10b</b>) was designed, synthesized, and structurally characterized using IR, ¹H/¹³C NMR, and mass spectrometry, and evaluated as dual-function antiproliferative and anti-fowlpox virus agents supported by integrated computational analyses. The synthesized compounds were screened for in vitro antiproliferative activity against HeLa, HCT-116, and MCF-7 cancer cell lines, as well as normal WI-38 lung fibroblasts. Several derivatives exhibited potent cytotoxic activity with enhanced selectivity toward cancer cells. Compound <b>5b</b> showed the highest activity against HeLa cells, compound <b>1</b> was most effective against HCT-116 cells, while compounds <b>7</b> and <b>8</b> displayed remarkable activity against MCF-7 cells, with compound <b>7</b> surpassing doxorubicin and compound <b>8</b> demonstrating excellent selectivity toward normal cells. Mechanistic investigations revealed that compounds <b>7</b> and <b>8</b> acted as dual topoisomerase I/IIβ inhibitors, inducing G2/M cell cycle arrest and intrinsic apoptosis associated with caspase-9 activation and downregulation of topoisomerase II protein expression. Selected derivatives were further evaluated for antiviral activity against fowlpox virus using in ovo and in vivo SPF embryonated chicken egg models, where compounds <b>2</b> and <b>9a</b> exhibited the highest therapeutic indices, comparable to ribavirin, and compound <b>9a</b> markedly suppressed viral replication and titers in vivo. ADMET profiling, molecular docking, molecular dynamics simulations, and DFT calculations supported the experimental findings and identified compound <b>10a</b> as the most favorable theoretical candidate. Overall, this integrated experimental-computational approach establishes naphthyridine derivatives as a rationally designed multifunctional chemotype for simultaneous anticancer and antiviral drug development.