The synthesis of triazinthions and their reactions with some nucleophilic reagents have been investigated during this scientific study. Thus, thiourea with a single component has been synthesized as a result of concomitant reactions of aldehyde and amines trials. The structure of the synthesized compounds was confirmed by 1H, 13C NMR spectroscopy methods. The inhibitory effects of novel N‐substituted triazinane‐2‐thione derivatives on acetylcholinesterase (AChE) activity were performed according to the spectrophotometric method of Ellman et al. These novel N‐substituted triazinane‐2‐thiones derivatives were effective inhibitors of the α‐glycosidase, cytosolic carbonic anhydrase I and II isoforms (hCA I and II), and Acetylcholinesterase (AChE) enzymes with Ki values in the range of 1.01 ± 0.28 to 2.12 ± 0.37 nm for α‐glycosidase, 13.44 ± 4.39 to 74.98 ± 6.25 nm for hCA I, 10.41 ± 4.8 to 72.6 ± 17.66 nm for hCA II, 36.82 ± 9.95 to 108.48 ± 1.17 nm for AChE, and 624.62 ± 100.34 to 1124.16 ± 205.14 nm for α‐glycosidase, respectively.