Protection of enkephalins from enzymatic degradation utilizing selective metal-chelating inhibitors

Metal ion-chelating agents inhibited enkephalin degradation by a rat striatal membrane-associated endopeptidase termed 'enkephalinase'. The combination of a hydrophobic dipeptidyl moiety and a transition metal-chelating moiety in the same molecule resulted in very efficient and selective inhibitors of enkephalinase. The mercaptoacetyl dipeptides (2-mercaptoacetyl-Leu-Phe and 2-mercaptoacetyl-Phe-Leu) and the N-phosphorylated dipeptides (phosphoryl-Leu-Phe and phosphoramidon) inhibited enkephalinase with IC50 values of 15, 70, 0.3 and 1 nM respectively, but were much less potent against the aminopeptidase and angiotensin converting enzyme, two other metalloenzymes implicated in the degradation of the enkephalins in brain. The inhibition of enkephalinase, using phosphoryl-Leu-Phe as a selective inhibitor, resulted in a 4 fold increase in the amount of enkephalin recovered following K+ depolarization of rat striatal slices.