Published Paper Details:

Chlorambucil, a well-established alkylating agent, has long been used in the treatment of chronic lymphocytic leukemia (CLL); however, its clinical efficacy is often limited by drug resistance mechanisms, including DNA repair and glutathione-mediated detoxification. To overcome these limitations and enhance therapeutic potential, we report the rational design, synthesis, and biological evaluation of a novel series of chlorambucil-based hybrid molecules capable of dual inhibition of DNA and histone deacetylases (HDACs). These bifunctional compounds are engineered to retain the DNA-alkylating ability of chlorambucil while incorporating HDAC inhibitory moieties to induce epigenetic modulation and sensitize tumor cells to DNA damage.The hybrids were synthesized via a modular approach and structurally characterized by NMR and mass spectrometry. Their cytotoxicity was evaluated in vitro using leukemia cell lines and primary CLL cells. HDAC inhibition was confirmed by increased acetylation of histone H3 and H4, while DNA alkylation and cross-linking were quantified using comet assays and ?-H2AX staining. Furthermore, glutathione (GSH) levels and glutathione S-transferase (GST) activity were measured to assess cellular detoxification responses. Notably, the novel hybrids induced robust DNA damage and apoptosis even in samples with elevated GSH and GST, indicating partial circumvention of resistance pathways.These findings support the therapeutic promise of dual-targeting chlorambucil derivatives as more effective anticancer agents, especially in hematologic malignancies with established resistance to monofunctional drugs. Further in vivo studies are warranted to evaluate pharmacokinetics and therapeutic efficacy.