This study investigates the synthesis and biological evaluation of 1,2,4-triazine-sulfonamide hybrids (3a–r) as potential inhibitors of acetylcholinesterase (AChE) and glutathione S-transferase (GST). The hybrids were synthesized through a one-step procedure involving the reaction of methylsulfonyl-5,6-diphenyl-1,2,4-triazine with various sulfonamides. The synthesized compounds were evaluated for their inhibitory effects on AChE and GST activities, both in silico and in vitro. The IC50 values for AChE inhibition ranged from 1.674 to 5.059 μM, with some hybrids showing enhanced inhibitory activity compared to the standard compound tacrine. For GST inhibition, the IC50 values ranged from 1.608 to 4.880 μM, with specific hybrids demonstrating favorable inhibition results. Molecular docking studies were conducted to understand the binding modes and interactions of the hybrids with AChE and GST, revealing key amino acid residues involved in the binding process. ADMET predictions were also performed to assess the pharmacokinetic properties of the hybrids, indicating their potential for oral administration and low toxicity. The study highlights the potential of these 1,2,4-triazine-sulfonamide hybrids as effective inhibitors of AChE and GST, warranting further development as therapeutic agents.This study investigates the synthesis and biological evaluation of 1,2,4-triazine-sulfonamide hybrids (3a–r) as potential inhibitors of acetylcholinesterase (AChE) and glutathione S-transferase (GST). The hybrids were synthesized through a one-step procedure involving the reaction of methylsulfonyl-5,6-diphenyl-1,2,4-triazine with various sulfonamides. The synthesized compounds were evaluated for their inhibitory effects on AChE and GST activities, both in silico and in vitro. The IC50 values for AChE inhibition ranged from 1.674 to 5.059 μM, with some hybrids showing enhanced inhibitory activity compared to the standard compound tacrine. For GST inhibition, the IC50 values ranged from 1.608 to 4.880 μM, with specific hybrids demonstrating favorable inhibition results. Molecular docking studies were conducted to understand the binding modes and interactions of the hybrids with AChE and GST, revealing key amino acid residues involved in the binding process. ADMET predictions were also performed to assess the pharmacokinetic properties of the hybrids, indicating their potential for oral administration and low toxicity. The study highlights the potential of these 1,2,4-triazine-sulfonamide hybrids as effective inhibitors of AChE and GST, warranting further development as therapeutic agents.