Inhibition of topoisomerase II by antitumor agents bis(2,6-dioxopiperazine) derivatives.

ABSTRACT

Several recently developed derivatives of bis(2,6-dioxopiperazine) have been shown to be new antitumor agents and are currently under clinical trials. We found that the mother compound of the bis(2,6-dioxopiperazine)s, ICRF-154, and its derivatives, ICRF-159, ICRF-193, and MST-16, are all inhibitors of mammalian type II DNA topoisomerase. By decatenation assay using kinetoplast DNA from Crithidia fasciculata, inhibition of purified calf thymus topoisomerase II by these compounds was investigated. Potency of inhibition was in the following order ICRF-193 greater than ICRF-154 = ICRF-159 greater than MST-16. The doses giving 50% inhibition were 2, 13, 30 and 300 microM, respectively, for these compounds. ICRF-193, the most potent inhibitor, however, did not inhibit topoisomerase I at concentrations up to 300 microM. Addition of excess enzyme, but not of the substrate DNA, overcame the inhibition by ICRF-193. The drug did not stimulate the formation of cleavable complex between DNA and the enzyme. Furthermore, ICRF-193 even inhibited the formation of enzyme-mediated DNA cleavage induced by etoposide or 4'-[9-acridinylamino)methanesulfon-m-anisidide. These observations, together with the finding that ICRF-193 did not intercalate into DNA, suggest that ICRF-154 and related compounds are specific inhibitors of topoisomerase II with different modes of action i.e., they interfere with some step(s) before the formation of the intermediate cleavable complex in the catalytic cycle. This is a property quite distinct from previously known cleavable complex-forming type topoisomerase II-targeting antitumor agents such as acridines, anthracyclines, and epipodophyllotoxins, but rather, mechanistically similar to the recently reported group of inhibitors that includes merbarone, aclarubicin, and fostriecin.