IODO AND DIIODOTYROSINE EPOXYSUCCINYL DERIVATIVES AS SELECTIVE INHIBITORS OF CATHEPSIN-B

Eight new analogs of -trans-epoxysuccinyl-L-leucylamido(3-methyl)butan e (E-64-c) containing Phe, Tyr, Tyr(I) or Tyr(I-2) in place of Leu, we re synthesized and tested as inhibitors of papain, bovine spleen cathe psin B, calpain I and II from porcine red cells and porcine kidney, re spectively. By use of kinetic methods, the new E-64 analogs proved to irreversibly inactivate both papain and cathepsin B via reversible enz yme-inhibitor intermediates EI. Second-order rate constants for inacti vation were in the range 3500-55 100 M(-1)s(-1) for papain and 650-105 000 M(-1)s(-1) for cathepsin B. For the inactivation of calpain I and II they ranged between 250 and 2000 M(-1)s(-1) and were similar to th ose of the known E-64-c. The effectiveness of the amino acid contained in the inhibitors tested increased in the order Tyr(I) approximate to Tyr(I-2) < Tyr < Phe < Leu for papain and Phe < Tyr < Tyr(I)e Leu < T yr(I-2) for cathepsin B inactivation. Replacement of the L with the D- trans-epoxysuccinyl unit caused a 10-100-fold decrease in inhibitor po tencies.