Inhibition of class A and C beta-lactamases by diaroyl phosphates.

A series of diaroyl phosphates was employed to assess the general reactivity of this class of molecule against classical class A and class C beta-lactamases. The compounds were found, in general, to be inhibitory substrates of both classes of enzyme. In each case, they reacted rapidly with the enzyme (10(4) to 10(6) s(-1) M(-1)) to yield transiently stable intermediates, most likely acyl-enzymes, which slowly (10(-3) to 10(-1) s(-1)) regenerated free enzyme. In certain cases, side branches from direct turnover produced EII complexes ("substrate" inhibition), more inert EI' complexes, and, in one case, a completely inactive EI' complex. Deacylation, but not acylation, was enhanced by electron-withdrawing substituents. Acylation rates were enhanced by hydrophobic substitution, both in the diaroyl phosphate and at the enzyme active site. The latter factor led to the general order of beta-lactamase acylation rates: class D (previous results) > class C > class A. It is likely that nanomolar inhibitors of all serine beta-lactamases could be achieved by rational exploitation of diacyl phosphates.