In vitro inhibition of Pseudomonas aeruginosa elastase by metal-chelating peptide derivatives

Efrat Kessler, Mary Israel, Nahum Landshman, Aaron Chechick, Shmaryahu Blumberg

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Pseudomonas aeruginosa elastase is a zinc metalloendopeptidase, probably responsible for the tissue destruction observed during infections with this organism. The elastase of a virulent Pseudomonas aeruginosa strain (Habs serotype 1) was isolated and found to have a molecular weight of 35,000; it readily degraded elastin and cartilage proteoglycans. A series of amino acid and peptide derivatives containing the metal-chelating moieties hydroxamate, phosphoryl, or thiol were synthesized and tested as potential inhibitors of the enzyme. Inhibition constants (K(iS)) for the compounds were determined with the chromophoric substrate furylacryloyl-glycyl-L-leucyl-L-alanine. The hydroxamic acid derivatives of benzyloxycarbonyl-glycine, benzyloxycarbonyl-L-leucine and benzyloxycarbonyl-L-phenylalanine had inhibition constants in the range of 11 to 28 μM. The 2-mercaptoacetyl derivatives of L-leucyl-D-phenylalanine and L-leucyl-L-phenylalanine had K(i) values of 34 and 1.5 μM, respectively, demonstrating the stereospecificity of the inhibition. The most potent inhibitors tested were 2-mercaptoacetyl-L-phenylalanyl-L-leucine and phosphoryl-L-leucyl-L-phenylalanine (K(i) = 0.2 μM). Similar compounds lacking the metal-chelating moiety were about 3 orders of magnitude poorer inhibitors. When the inhibition of the enzyme activity towards azocasein, elastin, or cartilage was examined, inhibitor concentrations approximately 50-fold higher than the respective K(iS) were required to obtain 60 to 90% inhibition. Virtually complete inhibition was achieved with these substrates at inhibitor concentrations 500-fold higher than the respective K(iS) (0.1 to 14 mM). Although, 2-mercaptoacetyl-L-phenylalanyl-L-leucine and phosphoryl-L-leucyl-L-phenylalanine exhibited the same affinity to the enzyme, the latter was inferior in inhibiting cartilage proteolglycan degradation. 2-Mercaptoacetyl-L-phenylalanyl-L-leucine represents a class of potent elastase inhibitors that might prove useful in the management of P. aeruginosa infections.

Original languageEnglish
Pages (from-to)716-723
Number of pages8
JournalUnknown Journal
Issue number2
StatePublished - 1982

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