Autocatalytic inactivation of cytochrome p-450 and chloroperoxidase by 1-aminobenzotriazole and other aryne precursors

Abstract

Cytochrome P-450, as reported previously is inactivated during catalytic turnover of 1-aminobenzotriazole due to alkylation of its prosthetic heme group. NMR analysis of the heme adduct after removal of the iron atom identifies it unequivocally as a derivative of protoporphyrin IX in which two of the nitrogens are bridged by a benzene ring. Cytochrome P-450 destructive activity is retained by analogues with Me or Ac substituents on the exocyclic N but is lost when the N itself is removed or is replaced by a hydroxyl or nitro function. Prosthetic heme alkylation also occurs with 1-amino-1H-naphtho (2,3-d)triazole, the analogue with one additional benzene ring. In vivo studies suggest that 1-aminobenzotriazole is relatively nontoxic. Catalytic turnover of 1-aminobenzotriazole by chloro-peroxidase results in the formation of phenol and in inactivation of the enzyme. The phenol obtained in deuterated water incorporates one deuterium into the aromatic ring. The data indicate that benzyne, formed by enzymatic oxidation of 1-aminobenzotriazole, is responsible for inactivation of cytochrome P-450 and chloroperoxidase.