Synthetic inhibitors of carboxypeptidase N.

Abstract

Further to explore the functions of carboxypeptidase N (CPN) in vivo, we undertook two studies to find CPN inhibitors of high potency and relatively long duration of action. In each study we examined for inhibition of hydrolysis of [3H]benzoyl-Ala-Arg using pure bovine serum CPN or human serum. In the first such study we synthesized a series of acyl amino acids and acyl di - and tripeptides containing arginine, lysine or both. All proved to be weak inhibitors (Ki = 10(-3) to 10(-4) M). N alpha-carbamoyl-Arg was the strongest: Ki = 3.5 X 10(-5) M. In the second study we prepared S-acyl (thio ester) derivatives of the highly potent CPN inhibitor 2-mercaptomethyl-3-guanidinoethylthiopropionic acid (2-MGP), as certain S-acyl groups markedly increase the duration of captopril, another mercapto-containing compound. Acetyl-, Boc-phenylalanyl-, phenylalanyl-, benzoyl-alanyl-, alanyl-, and Boc-alanyl-2-MGP retained the high potency of 2-MGP in vitro. Although Ala-2-MGP exerted maximum effects in vivo, like those of 2-MGP, the duration of action of Ala-2-MGP was slightly shorter than that of 2-MGP. These results indicate that the mercapto group of 2-MGP can be taken up in some forms of thioester linkage and still remain virtually the full potency of 2-MGP itself. Thus, it appears that a free mercapto function is not essential for the action of 2-MGP.