AMINOOXYPHOSPHONATES AS SLOW BINDING INHIBITORS OF ASPARTATE AND ALANINE AMINOTRANSFERASES FROM PORCINE HEART

Aminooxymethylphosphonic (AOMP), 1-aminooxyethylphosphonic (1-AOEP) and 2-aminooxyethyl-phosphonic (2-AOEP) acids have been synthesised and were found to be potent slow binding inhibitors of aspartate- and alanine-aminotransferases with K-i ranging from nanomolar to micromolar values. The half-life of the inhibited complexes varied from 8 min (AspAT-2-AOEP) to 11 h (AspAT-AOMP). Kinetic analysis of the interaction of both enzymes with AOMP suggested the formation of an E-I complex in a single slow binding process. In the case of other compounds, attempt to discriminate between a single- or a double-step mechanism, consistent with an EI intermediate followed by a slow E-I to E-I* isomerisation process, could not be clearly resolved. Spectral studies of the complex formed between PLP-bound enzyme and the aminooxy compound resulted in a shift from 362 nm, the absorption maximum of the native enzyme, to 380 nm, characteristic of the oxime produced. The kinetic parameters for aminooxyphosphonates were compared to those for their carboxylic and aminophosphonic analogues.