In this work, three new molecules 4-hydroxy salicyaldehyde ethane sulfonylhydrazone (4-OH-salesh), 4-hydroxy salicylaldehyde propane sulfonylhydrazone (4-OH-salpsh) and 4-hydroxy salicylaldehyde butane sulfonylhydrazone (4-OH-salbsh) are synthesized by combining C2H5/C3H7/C4H9 sulfonyl chlorides and hydrazine hydrate and 4-hydroxysalicylaldehyde. The synthesized compounds were characterized by using FT-IR spectroscopic technique. In addition, DFT analyzes of these compounds were performed.Also, three alkyl sulfonylhydrazon series and choline esterase enzymes tested to determine their inhibition potential against the acetylcholinesterase (AChE) and butyrylcholinesterase BChE enzymes. in vitro results showed that the compounds could act as potent inhibitors of AChE and BChE. The compound (4-OH-salbsh) showed the most potent inhibitory property against three of the enzymes with IC50 values of 12 µM for AChE, 23 µM for BChE.