In silico and in vitro evaluation of newly synthesized pyrazolo-pyridine fused tetrazolo-pyrimidines derivatives as potential anticancer and antimicrobial agents

Abstract

Diversely functionalized pyrazolo-pyridine fused tetrazolo-pyrimidines 10aa-am and 10ba-bn were successfully synthesized via a catalyst-free synthetic protocol with moderate to very good yields. The compounds were evaluated for cytotoxicity against MCF-7 and HEK-293 cells using MTT assay. Among the tested compounds, 10ab (IC₅₀- 23.83 µM) and 10ah (IC₅₀- 23.30 µM) demonstrated the highest potency against MCF-7 cells, while 10bc (IC₅₀- 14.46 µM) and 10bh (IC₅₀- 2.53 µM) exhibited excellent cytotoxicity against HEK-293 cells. Additionally, antibacterial screening was performed against three Gram-negative bacteria (E. coli, P. aeruginosa, and S. enterica) and three Gram-positive bacteria (S. aureus, B. megaterium, and B. subtilis) using broth dilution method, while antifungal activity was assessed against three fungal strains (A. niger, Penicillium, and S. cerevisiae) using agar well diffusion method. In antimicrobial screening, the majority of the compounds demonstrated significant antibacterial efficacy compared to antifungal activity. We also conducted comprehensive computational studies, including DFT calculations, molecular docking and dynamics, and drug-likeness assessments. In the DFT study, compounds 10ac and 10bc displayed stable conformations, indicating their potential for higher therapeutic activity. Molecular docking analyses revealed compelling interactions, with compound 10ah demonstrating docking score −7.42 kcal/mol against catalytical domain PARP1 (PDB ID: 7KK4) and 10bh exhibiting a best docking score −10.77 kcal/mol against human corticotropin-releasing factor receptor 1 (PDB ID: 4Z9G). A 100 ns molecular dynamics (MD) simulation study of compounds 10ah and 10bh revealed the stable conformation and binding energy in a stimulating environment. In drug-likeness assessments, both the compounds 10ah and 10bh adhere all the established guidelines.

Communicated by Ramaswamy H. Sarma

Keywords:

• Pyrazolo-pyridine fused tetrazolo-pyrimidine
• catalyst-free synthesis
• cytotoxicity
• antimicrobial
• density functional calculations
• molecular docking and dynamics

Acknowledgements

Authors HCP, MSP, JNP, DDC and PD are indebted to Knowledge Consortium of Gujarat (KCG), Department of Education, Government of Gujarat, India for SHODH-Scheme fellowship. Authors are thankful to Centre with Potential for Excellence in Particular Area (CPEPA, Phase-II) with award letter No. F.No.1-14/2002-2016 (NS/PE) dated 28^th April, 2016 and Centre for Advance Study (CAS, Phase-II) with award letter No. F-540/5/CAS-II/2018 (SAP-I), dated 25^th January, 2018 for providing essential chemicals and basic instrumental facility. Authors are also thankful to the Head, Department of Chemistry, Sardar Patel University for providing basic amenities for carrying out this work. The authors are thankful to Dr. Alan for providing computational resources by the e-INFRA CZ project (ID:90140), supported by the Ministry of Education, Youth and Sports of the Czech Republic.

Disclosure statement

Authors declare no competing financial interest.

Additional information

Funding

The author(s) reported there is no funding associated with the work featured in this article.