Design and synthesis of novel multi-target tetrabromophthalimides as CBS and Topo-II inhibitors and DNA intercalatorsElectronic supplementary information (ESI) available: GI% of tetrabromophthalimide derivatives (2a–2k) utilizing fifteen cancer and two normal cell lines (Table S1); some reported β-tubulin CBS inhibitors, Topo-II poisons (doxorubicin and mitoxantrone), and Topo-II catalytic inhibitors (etoposide) (Fig. S1); FT-IR, 1H and 13C NMR, and mass spectra of all candidates (Fig. S2–S31); biological data (Fig. S32–S41) and materials and methods (SI1–SI6). See DOI: https://doi.org/10.1039/d4md00585f

Microtubules are highly dynamic structures and constitute a crucial component of the cellular cytoskeleton. Besides, topoisomerases (Topo) play a fundamental role in maintaining the appropriate structure and organization of DNA. On the other hand, dual mechanism drug candidates for cancer treatment primarily aim to enhance the efficacy of cancer treatment and potentially overcome drug resistance. Hence, this work was tailored to design and synthesize new multi-target tetrabromophthalimide derivatives (2a–2k) that are capable of inhibiting the colchicine binding site (CBS) and topoisomerase II (Topo-II). The conducted in vitrostudies showed that compound 2fshowed the lowest IC50value (6.7 μg mL−1) against the MDA-MB-468 cancer cell line. Additionally, compound 2fprompted upregulation of pro-apoptotic markers (caspases 3, 7, 8, and 9, Bax and p53). Moreover, some anti-apoptotic proteins (MMP2, MMP9, and BCL-2) were downregulated by compound 2ftreatment. Besides, the colchicine binding assay showed that compounds 2fand 2kdisplayed promising inhibitory potential with IC50values of 1.92 and 4.84 μg mL−1, respectively, in comparison with colchicine (1.55 μg mL−1). Furthermore, the Topo-II inhibition assay displayed the prominent inhibitory potential of compound 2fwith an IC50value of 15.75 μg mL−1, surpassing the IC50of etoposide (20.82 μg mL−1). Cell cycle analysis revealed that compound 2finduced cell cycle arrest at both the G0–G1 and G2–M phases. The new candidates were docked against both the CBS (PDB ID: 5XIW) and Topo-II (PDB ID: 5CDP) targets to investigate their binding interactions and affinities as well. Accordingly, the synthesized compounds could serve as promising multi-target anticancer candidates with eligible apoptotic activity.