PubMed1999
Alkyl-N-phenyl carbamates (2-8) (see Figure 1), alkyl-N-phenyl thiocarbamates (9-15), 2,2'-biphenyl-2-ol-2'-N-substituted carbamates (16-23), and 2, 2'-biphenyl-2-N-octadecylcarbamate-2'-N-substituted carbamates (24-31) are prepared and evaluated for their inhibition effects on porcine pancreatic cholesterol esterase and Pseudomona species lipase. All inhibitors are characterized as transient or pseudo substrate inhibitors for both enzymes. Both enzymes are not protected from inhibition and further inactivated by carbamates 2-8 and thiocarbamates 9-15 in the presence of trifluoroacetophenone. Therefore, carbamates 2-8 and thiocarbamates 9-15 are exceptions for active site binding inhibitors and are probably the second alkyl-chain binding-site-directed inhibitors for both enzymes. The inhibition data for carbamates 2-8 and thiocarbamates 9-15 are correlated with the steric constant, E(s), and the hydrophobicity constant, pi; however, the inhibition data are not correlated with the Taft substituent constant, sigma. A comparison of the inhibition data for carbamates 2-8 and thiocarbamates 9-15 toward both enzymes indicates that thiocarbamates 9-15 are more potent inhibitors than carbamates 2-8. A comparison of the inhibition data for cholesterol esterase and Pseudomona species lipase by carbamates 2-8 or thiocarbamates 9-15 indicates that cholesterol esterase is more sensitive to the E(s) and pi values than Pseudomona species lipase. The negative slope values for the logarithms of inhibition data for Pseudomona species lipase by carbamates 2-8 and thiocarbamates 9-15 versus E(s) and pi indicate that the second alkyl-chain-binding site of Pseudomona species lipase is huge, hydrophilic, compared to that of cholesterol esterase, and prefers to interact with a bulky, hydrophilic inhibitor rather than a small, hydrophobic one. On the contrary, the second alkyl-chain-binding site of cholesterol esterase prefers to bind to a small, hydrophobic inhibitor. Both enzymes are protected from inhibition by carbamates 16-23 in the presence of trifluoroacetophenone. Therefore, carbamates 16-23 are characterized as the alkyl chain binding site, esteratic site oxyanion active site directed pseudo substrate inhibitors for both enzymes. Both enzyme inhibition data for carbamates 16-22 are well-correlated with sigma alone. The negative rho values for these correlations indicate that the serine residue of both enzymes and carbamates 16-22 forms the tetrahedral species with more positive charges than inhibitors and the enzymes and follow the formation of the carbamyl enzymes with more positive charges than the tetrahedral species. Carbamates 24-31 are also exceptions for active site binding inhibitors and probably the second alkyl chain binding site-directed inhibitors for both enzymes. However, the enzyme inhibition constants for carbamates 24-31 are correlated with values of sigma, E(s), and pi. The negative rho values for these correlations indicate that both enzymes and carbamates 24-31 form the tetrahedral species with more positive charges than inhibitors and the enzymes and follow the formation of the carbamyl enzymes with more positive charges than those tetrahedral species. Therefore, carbamates 24-31 may bind to both the active sites and the second alkyl chain binding site and follow the evacuation of the active sites. A comparison of the rho values for cholesterol esterase and Pseudomona species lipase by carbamates 24-31 indicates that cholesterol esterase is much more sensitive to the sigma values than Pseudomona species lipase. The negative sensitivity values, delta, for the cholesterol esterase inhibitions by carbamates 24-31 indicate that the enzyme prefers to bind to a bulky carbamyl group rather than bind to a small one. The hydrophobicity of carbamates 24-31 does not play a major role in both enzyme inhibitions.