Using computational methods of molecular modelling conformational changes in the active site of acetylcholinesterase (AChE; EC 3.1.1.7) upon binding of various ligands were investigated and structural characteristics of efficient oxime reactivators of AChE inhibited with warfare nerve agents and gudelines for their synthesis were defined. Four new chiral oxime reactivators from N-substituted 2-hydroxiiminoacetamide group were prepared starting from racemic 1-phenyl-allylamine prepared from cinnamyl alcohol. Enantiomers of oximes were separated using high performance liquid chromatography on polysaharidic chiral stationary phases. New oximes were tested for inhibition of AChE and butyrylcholinesterase (BChE; EC 3.1.1.8) and reactivation of cholinesterases inhibited with tabun, cyclosarin, sarin and VX. New oximes reversibly inhibit both enzymes with inhibition constant (KI) in the micromolar range. Both enzymes showe greatest affinity toward 2-hydroxyimino-N-(3-(4-((2- methylimidazol-1-yl)methyl)-1,2,3-triazol-1-yl)-1-phenylpropyl)acetamide towards which BChE displays significant selectivity and stereoselectivity. Only this oxime showed greater reactivation efficiency than the reference oximes described in literature in the case of BChE inhibited with cyclosarin. Molecular docking studies concluded that differences in binding of new oximes in AChE and BChE largely result from differences in the amino acids at the position of Tyr72, Tyr124, Phe297 and Tyr337 in the AChE active site.