Benzodioxin-Annulated Naphthalimides as Potent DNA Replication Stress Inducers with Dual p53-Dependent and Independent Antitumor Activity

TL;DR

This study introduces a new compound that causes DNA replication stress and shows strong antitumor activity in cancer cells.

Contribution

A novel naphthalimide derivative (compound 5a) is identified as a potent DNA replication stress inducer with dual p53-dependent and independent antitumor activity.

Findings

Compound 5a shows strong cytotoxic effects in tumor cells with favorable selectivity over non-malignant cells.

Treatment with 5a causes DNA synthesis inhibition and induces DNA damage markers like γH2AX.

Compound 5a leads to cell-cycle arrest and apoptosis, indicating irreversible long-term cellular damage.

Abstract

Background/Objectives: The development of small-molecule agents that selectively target DNA replication remains a central strategy in anticancer drug discovery. In this study, we report the biological characterization of a novel 6-nitro-benzodioxin-naphthalimide (NI) derivative (compound 5a), evaluated as a potential DNA-targeted anticancer lead. Methods/Results: The antiproliferative activity of 5a was assessed in a small panel of human lung carcinoma cell models (A549, H1299) and a non-malignant control (MRC-5), revealing pronounced cytotoxic effects in tumor cells, accompanied by favorable selectivity indices. Mechanistic investigations demonstrated that treatment with 5a results in strong inhibition of DNA synthesis, as evidenced by a marked reduction in EdU incorporation and a robust induction of the DNA damage marker γH2AX. These effects were associated with cell-cycle perturbations characterized by accumulation in G1 and G2/M phases, followed by activation of apoptotic pathways. Importantly, clonogenic survival assays confirmed that even transient exposure to 5a leads to a sustained loss of proliferative capacity, indicating irreversible long-term cellular damage. These results support a replication stress-driven mechanism of action for compound 5a, consistent with interference in DNA-associated processes during S phase. Conclusions: While the precise molecular initiating event remains to be elucidated, the observed biological profile positions 5a as a promising DNA-targeted lead structure with potential for further pharmaceutical optimization. These findings provide a solid foundation for the continued development of naphthalimide-based compounds as anticancer agents within a pharmaceutically relevant framework.

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