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Benzimidazole-linked pyrazolo[1,5-a]pyrimidine conjugates: synthesis and detail evaluation as potential anticancer agents

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Abstract

A library of benzimidazole briged pyrazolo[1,5-a]pyrimidine (6a-q) was designed, synthesized and subjected for evaluation for cytotoxic potential. Antiproliferative activity, ranging from 3.1–51.5 μM, was observed against a panel of cancer cell lines which included MCF-7 (breast cancer), A549 (lung cancer), HeLa (cervical cancer) and SiHa (cervical cancer). Among them, 6k, 6l, 6n and 6o have shown significant cytotoxicity and were investigated further to study their probable mechanism of action against MCF-7 cell line. Accumulation of cells at sub-G1 phase was observed in flow cytometric analysis. The detachment of cells from substratum and membrane blebbing seen under bright field microscopy supports the ability of these conjugates to induce apoptosis. Immunostaining and western blot analysis showed EGFR, p-EGFR, STAT3, and p-STAT3 significant downregulation. Western blot analysis demonstrated an elevated level of apoptotic proteins such as p53, p21, Bax, whereas a decrease in the antiapoptotic protein Bcl-2 and procaspase-9, confirming the ability of these conjugates to trigger cell death by apoptosis. EGFR kinase assay confirms the specific activity of conjugates. Molecular docking simulation study disclosed that these molecules fit well in ATP-binding pocket of EGFR. The analysis of docking poses and the atomic interactions of different conjugates rationalize the structural–activity relationship in this series.

Graphical abstract

Benzimidazole-linked pyrazolo[1,5-a]pyrimidine conjugates were synthesized and evaluated for their anticancer potential. All the conjugates have significant anticancer potential. Further mechanistic studies revealed that these conjugates arrest cancer cell growth by EGFR/STAT3 inhibition.

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Acknowledgements

The author C.B acknowledges the Department of Pharmaceuticals (Ministry of Chemicals and Fertilizers, Govt. of India) for the financial support & the CSIR-IICT, Hyderabad for providing facilities. K.R.G thanks UGC for his fellowship. This work was financially supported by the CSIR 12th FYP CSC0111 (SMiLE). The authors thank Y. Suresh for the Flow Cytometry and Confocal Microscopy studies.

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Authors and Affiliations

  1. Department of Medicinal Chemistry, National Institute of Pharmaceutical Education and Research (NIPER), Hyderabad, 500 037, India

    Chandrakant Bagul, Ravikumar Akunuri & Ahmed Kamal

  2. Medicinal Chemistry and Pharmacology, CSIR-Indian Institute of Chemical Technology, Hyderabad, 500 007, India

    Chandrakant Bagul, Jaki R. Tamboli, Siddiq Pasha Shaik & Ahmed Kamal

  3. Department of Pharmaceutical Chemistry, Amrita School of Pharmacy, Amrita Vishwa Vidyapeetham, AIMS Health Sciences Campus, Kochi, 682 041, India

    Chandrakant Bagul

  4. Chemical Biology, CSIR-Indian Institute of Chemical Technology, Hyderabad, 500 007, India

    Garikapati Koteswara Rao, Immadi Veena & Manika Pal-Bhadra

  5. Bharati Vidyapeeth’s Poona College of Pharmacy, Paud Road, Erandawane, Pune, 411038, India

    Ravindra Kulkarni

  6. School of Pharmaceutical Education and Research (SPER), Jamia Hadard, New Delhi, 110062, India

    Ahmed Kamal

  7. Birla Institute of Technology and Science, Hyderabad Campus, Pilani, 500078, India

    Ahmed Kamal

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Bagul, C., Rao, G.K., Veena, I. et al. Benzimidazole-linked pyrazolo[1,5-a]pyrimidine conjugates: synthesis and detail evaluation as potential anticancer agents. Mol Divers 27, 1185–1202 (2023). https://doi.org/10.1007/s11030-022-10481-x

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