Synthesis and antiproliferative evaluation of novel 3,5,8-trisubstituted coumarins against breast cancer

Abstract

Aim: This study focused on designing and synthesizing novel derivatives of 3,5,8-trisubstituted coumarin. Results: The synthesized compounds, particularly compound 5, exhibited significant cytotoxic effects on MCF-7 cells, surpassing staurosporine, and reduced toxicity toward MCF-10A cells, highlighting potential pharmacological advantages. Further, compound 5 altered the cell cycle and significantly increased apoptosis in MCF-7 cells, involving both early (41.7-fold) and late stages (33-fold), while moderately affecting necrotic signaling. The antitumor activity was linked to a notable reduction (4.78-fold) in topoisomerase IIβ expression. Molecular modeling indicated compound 5‘s strong affinity for EGFR, human EGF2 and topoisomerase II proteins. Conclusion: These findings highlight compound 5 as a multifaceted antitumor agent for breast cancer.

Summary points

• Novel derivatives of 3,5,8-trisubstituted coumarin and azacoumarin were synthesized and comprehensively characterized.

• Evaluation of the synthesized compounds against MCF-7 breast cancer cells through diphenyltetrazolium bromide assay revealed significant cytotoxic activity.

• Compound 5 demonstrated potent antiproliferative activity (IC₅₀ = 11.36 ± 0.55 μM) against MCF-7 cells, exceeding staurosporin, and exhibited lower toxicity (IC₅₀ = 39.70 μM) toward normal breast MCF-10A cells, suggesting pharmacological advantages.

• Flow cytometric analysis of the MCF-7 cell cycle highlighted alterations in cell population distribution, reducing cells in the G₀/G₁ and S phases while significantly increasing those in the G₂/M phase.

• Annexin V-FITC/propidium iodide analysis revealed substantial increases in both early and late-stage apoptosis in MCF-7 cells induced by compound 5, with a moderate impact on necrosis.

• Molecular mechanism investigation identified a significant reduction (4.78-fold) in the expression levels of topoisomerase IIβ in MCF-7 cells, associating it with compound 5's antitumor activity.

• Molecular modeling and dynamic studies showcased compound 5's considerable affinity for the binding pockets of EGFR, human EGF2 and topoisomerase II proteins.

Keywords: :

• breast cancer
• cell cycle distribution
• cytotoxic activity
• flow cytometric analysis
• molecular dynamics
• molecular modeling
• programmed cell death
• substituted coumarins
• topoisomerase IIβ

Author contributions

Conceptualization: MG Salem, SN Mali, EM Saied and MF Youssef; methodology: MG Salem, SN Mali, RD Jawarkar, SZ Alshawwa, E Al-Olayan, EM Saied and MF Youssef; software: AM Alqahtani, SN Mali, HA Alshwyeh, RD Jawarkar, AS Altamimi, EM Saied and MF Youssef; validation: MG Salem, AM Alqahtani, SN Mali, RD Jawarkar, AS Altamimi, SZ Alshawwa, E Al-Olayan, EM Saied and MF Youssef; formal analysis: MG Salem, SN Mali, HA Alshwyeh, RD Jawarkar, AS Altamimi, E Al-Olayan and MF Youssef; investigation: AM Alqahtani, HA Alshwyeh, RD Jawarkar, AS Altamimi, E Al-Olayan, EM Saied and MF Youssef; data curation: MG Salem, AM Alqahtani, SN Mali, HA Alshwyeh, RD Jawarkar, SZ Alshawwa and MF Youssef; writing – original draft preparation: MG Salem, AM Alqahtani, SN Mali, RD Jawarkar, EM Saied and MF Youssef; writing – review and editing: MG Salem, AM Alqahtani, SN Mali, HA Alshwyeh, RD Jawarkar, AS Altamimi, SZ Alshawwa, E Al-Olayan, EM Saied and MF Youssef; visualization: MG Salem, AM Alqahtani, HA Alshwyeh, AS Altamimi, EM Saied and MF Youssef; supervision: MG Salem, SN Mali, EM Saied and MF Youssef; project administration: MG Salem, SZ Alshawwa, E Al-Olayan, EM Saied and MF Youssef; funding acquisition: MG Salem, SZ Alshawwa, E Al-Olayan and EM Saied. All authors have read and agreed to the published version of the manuscript.

Financial disclosure

This research was funded by the Faculty of Pharmacy, Port-Said University, and Faculty of Science, Suez Canal University, Egypt. The authors also extend their appreciation to Princess Nourah bint Abdulrahman University for funding this work through the Researchers Supporting Project number PNURSP2023R165, Princess Nourah bint Abdulrahman University, Riyadh, Saudi Arabia. This study was also supported by Researchers Supporting Project number RSP2024R111, King Saud University, Riyadh, Saudi Arabia.

Competing interests disclosure

The authors have no competing interests or relevant affiliations with any organization or entity with the subject matter or materials discussed in the manuscript. This includes employment, consultancies, honoraria, stock ownership or options, expert testimony, grants or patents received or pending or royalties.

Writing disclosure

No writing assistance was utilized in the production of this manuscript.