Correlation of cytotoxicity and protein-associated DNA strand breaks for 2-(arylmethylamino)-1,3-propanediols.

Abstract

A mechanism of action study was performed with 14 novel DNA binding agents characterized structurally as 2-(arylmethylamino)-1,3-propanediols (AMAPs). Correlations between 8226 myeloma cell colony formation and DNA damage were performed using soft agar colony-forming assays and alkaline elution filter techniques respectively. The frequency of double-stranded breaks (DSBs), single-stranded breaks (SSBs) and DNA-protein cross-links were compared with cell growth inhibitory potency. Highly potent AMAPs in the colony formation assays included 91U86, an N-methyl-5-benzo(c)carbazole derivative, 773U82, a 3-substituted fluoranthene derivative, and crisnatol (770U82), the 6-substituted chrysene derivative. There was a high frequency of SSBs and DSBs with many analogues, but only SSBs occurred in a concentration-dependent fashion. Using regression analysis, the degree of single-strand damage correlated with cytotoxic potency for the AMAPs, with an R-value of 0.57 (P = 0.04). By gel electrophoresis assays, three clinically tested AMAPs, crisnatol BW 770U82, BW 502U83 and BW 773U82, were shown to inhibit the decatenation of pBR 322 DNA by purified topoisomerase-II (TOPO-II) enzymes. These results suggest that while some active AMAPs, such as crisnatol (BW 770U82), BW 502U83 and BW 773U82, inhibit TOPO-II enzymes, leading to protein-associated SSBs, other mechanisms, which do not involve DNA strand damage, must also contribute to the cytotoxic effects of this class of antitumor compounds. Intercalation has been well documented for these drugs and this may explain some of the growth inhibitory activity of the AMAPs.