The present work depicts synthesizing an innovative sequence of polyfunctionalized Pyrazolo[3,4-b]pyridine congeners 2-12 and assessed for antimicrobial, antitumor, telomerase inhibition, and anti-hepatitis B efficacies applying an array of techniques. Fortunately, most of the compounds demonstrated auspicious bio efficiencies. Additionally, in silico simulation was executed for determining the synthetic Pyrazolo[3,4-b]pyridines anticipated mode of action. Compounds 4, 5, and 6 were picked as the most potent and efficient candidates versus the telomerase enzyme [PDB id: 2B2A]. The consequences exposed that compound 5 is the extremely credible operative in contradiction of telomerase enzyme as it disclosed the best binding score and interaction pose. These initial outcomes may improve in introducing more research strategies converged on pyrazolo[3,4-b]pyridines, exclusively those with anticipated bioefficacy, ultimate ADMET parameters, and constructive prospects.