DESIGN, SYNTHESIS, DNA-SEQUENCE PREFERENTIAL ALKYLATION AND BIOLOGICAL EVALUATION OF N-MUSTARD DERIVATIVES OF HOECHST-33258 ANALOGS

The design, synthesis and biological evaluation of a series of bis-ben zimidazole analogues of Hoechst 33258 bearing nitrogen mustard moietie s is described. The novel compounds show clear evidence of interstrand cross-linking of linear lambda DNA, in contrast to their distamycin n itrogen mustard counterparts. Interference of the cross-linking reacti on by the minor groove-selective distamycin suggests that this process takes place in the minor groove of DNA. Sequence preferential alkylat ion is revealed by high-resolution polyacrylamide gel electrophoresis and autoradiography. Aikylation occurs predominantly at the 5'-A or 5' -G termini of mixed sequences, determined largely by the sequence-reco gnizing properties of the bis-benzimidazole carrier moiety. An analysi s of the frequency of bases around the alkylation sites reveals marked individual base preferences, especially for A at -3 and +3 positions. All of the compounds tested showed cytotoxic properties against the h uman tumor cell line KB in culture.