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Identification of bicyclic compounds that act as dual inhibitors of Bcl-2 and Mcl-1

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Abstract

Elevated expression of anti-apoptotic proteins, such as Bcl-2 and Mcl-1 contributes to poor prognosis and resistance to current treatment modalities in multiple cancers. Here, we report the design, synthesis and characterization of benzimidazole chalcone and flavonoid scaffold-derived bicyclic compounds targeting both Bcl-2 and Mcl-1 by optimizing the structural differences in the binding sites of both these proteins. Initial docking screen of Bcl-2 and Mcl-1 with pro-apoptotic protein Bim revealed possible hits with optimal binding energies. All the optimized bicyclic compounds were screened for their in vitro cytotoxic activity against two oral cancer cell lines (AW8507 and AW13516) which express high levels of Bcl-2 and Mcl-1. Compound 4d from the benzimidazole chalcone series and compound 6d from the flavonoid series exhibited significant cytotoxic activity (IC50 7.12 μM and 17.18 μM, respectively) against AW13516 cell line. Time Resolved-Fluorescence Resonance Energy Transfer (TR-FRET) analysis further demonstrated that compound 4d and compound 6d could effectively inhibit the Bcl-2 and Mcl-1 proteins by displacing their BH3 binding partners. Both compounds exhibited potent activation of canonical pathway of apoptosis evident from appearance of cleaved Caspase-3 and PARP. Further, treatment of oral cancer cells with the inhibitors induced dissociation of the BH3 only protein Bim from Mcl-1 and Bak from Bcl-2 but failed to release Bax from Bcl-xL thereby confirming the nature of compounds as BH3-mimetics selectively targeting Bcl-2 and Mcl-1. Our study thus identifies bicyclic compounds as promising candidates for anti-apoptotic Bcl-2/Mcl-1 dual inhibitors with a potential for further development.

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Acknowledgements

The authors would like to thank the following core facilities for their contributions to this work: Drug design and synthesis experiments were performed in the Institute of Chemical Technology, Department of Pharmaceutical Sciences and Technology; NMR and Computer Aided Drug Design Facility for instrumentation. This facility receives support from TEQIP Grant (TEQIP II/TEQIP-II/MH/MH2G01/386/347).

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  1. Abhay Uthale and Aarti Anantram have contributed equally to this work.

Authors and Affiliations

  1. Advanced Centre for Treatment, Research & Education in Cancer (ACTREC), Tata Memorial Centre, Kharghar, Navi Mumbai, 410 210, India

    Abhay Uthale, Prasad Sulkshane & Tanuja Teni

  2. Department of Pharmaceutical Sciences and Technology, Institute of Chemical Technology, Mumbai, 400 019, India

    Aarti Anantram & Mariam Degani

  3. Homi Bhabha National Institute, Training School Complex, Anushakti Nagar, Mumbai, 400085, India

    Abhay Uthale, Prasad Sulkshane & Tanuja Teni

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Uthale, A., Anantram, A., Sulkshane, P. et al. Identification of bicyclic compounds that act as dual inhibitors of Bcl-2 and Mcl-1. Mol Divers 27, 1359–1374 (2023). https://doi.org/10.1007/s11030-022-10494-6

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