4-Nitrophenyl-N-substituted carbamates (1) are characterized as pseudosubstrate inhibitors of acetylcholinesterase. The first step is formation of the enzyme-inhibitor tetrahedral intermediate with the inhibition constant (K i ), the second step is formation of the carbamyl enzyme with the carbamylation constant (k c ), and the third step is hydrolysis of the carbamyl enzyme with decarbamylation constant (k d ).
According to pre-steady state kinetics the K i step is divided further into two steps: (1) formation of the enzyme-inhibitor complex with the dissociation constant (K S ) and (2) formation of the enzyme-inhibitor tetrahedral intermediate from the complex with the equilibrium constant (k 2 /k -2 ). Since the inhibitors are protonated in pH 7.0 buffer solution, the virtual dissociation constant (K S ) of the enzyme-protonated inhibitor complex can be calculated from the equation, -log K S = -log K S -pK a + 14. The -log K S , -log K S , log k 2 , and log k -2 values are multiply linearly correlated with the Jäve equation (log(k/k 0 ) = * * + ␦E s + ). For -log K S - * -E s --correlation, the * value of -0.4 indicates that the enzymeprotonated inhibitor complexes have more positive charges than the protonated inhibitors, the ␦ value of 0.44 suggests that the bulkily substituted inhibitors lessen the reaction due to the difficulty of the inhibitors to enter the narrow enzyme active site gorge, and the value of 0.27 implies that the inhibitors with hydrophobic substituents accelerate the inhibitors entering the active site gorge of the enzyme. For log k 2 /k -2 - * -E s --correlation, the * value of 1.1 indicates that the enzyme-protonated inhibitor tetrahedral intermediates have more negative charges than the enzyme-protonated inhibitor complexes, the ␦ value of 0.15 suggests that the bulkily substituted inhibitors are difficult to bind into a small acyl binding site of