In this study, a series of novel sulfonamide derivatives (6–10) bearing an aryl sulfonate moiety was designed, and synthesized for the first time, and their inhibitory activities against human carbonic anhydrase isoenzymes (hCA II and I), acetylcholinesterase (AChE), and butyrylcholinesterase (BChE) were investigated by in vitro and silico studies. The chemical structures of the derivatives were elucidated by elemental analysis and spectral techniques. All tested hybrids showed remarkably low nanomolar inhibition with Kı values of in the range of 8.98 ± 0.10 to 14.56 ± 0.41 nM against hCA I, 7.73 ± 0.10 to 13.85 ± 0.32 nM against hCA II, 12.90 ± 0.29 to 16.38 ± 0.41 nM against AChE, and 6.21 ± 0.09 to 14.52 ± 0.38 nM against BChE. Overall, the majority of these molecules inhibited these metabolic enzymes significantly more than acetazolamide (AZA) and neostigmine. Moreover, the biological effects of the compounds produced were assessed through computational studies of target enzymes. In this regard, in silico ADMET properties were evaluated to determine the physicochemical properties of target compounds. The calculated results support the biological activity results.