In this study, it was aimed to synthesize (E)-N′-(2hydroxybenzylidene)-substituted benzenesulfonohydrazide (1−7) from the 2-hydroxybenzaldehyde reaction of different substituted fluorinated sulfonyl hydrazides. The structures of the synthesized molecules were characterized by elemental analysis, FTIR, 1 H NMR, 13 C NMR, 19 F NMR, and 2D NMR (HMBC, correlation spectroscopy, and HQSC). The anticholinesterase (AChE and BChE) and antidiabetic (α-glucosidase, α-amylase) inhibition activities of the synthesized compounds were evaluated. According to biological activity test results, (E)-N′-(2-hydroxybenzylidene)-4-(trifluoromethoxy)benzenesulfonohydrazide (compound 7 among hydrazone derivatives 1−7) demonstrated better BChE inhibitor activity than galantamine in anticholinesterase inhibition; and in the α-glucosidase and α-amylase assay, it exhibited more antidiabetic inhibition activity than the reference standard.