DNA crosslinking, sister chromatid exchange and cytotoxicity of N-2-chloroethylnitrosoureas tethered to minor groove binding peptides.

Abstract

Chloroethylnitrosoureas (CENU) are clinically important chemotherapeutic agents whose mechanism of action involves the formation of interstrand DNA crosslinks via an ethane bridge between N1-G and N3-C. CENU generally alkylate G at the N7- and O6-positions, with the latter lesion being the precursor to the interstrand crosslink. In previous studies, we reported the synthesis of CENU appended by a C2H4 linker to the N-terminus of DNA minor groove binding dipeptides (lex, information reading peptides) based on N-methylpyrrole-carboxamide subunits. Because of the dipeptide structure, these CENU-lex's react with DNA at adenines associated with lex equilibrium binding sites. No other CENU has been reported to yield A adducts. The biological evaluation of these CENU-lex's show that they are somewhat less cytotoxic than their simpler counterparts. In addition, in vitro studies show that the minor groove binding CENU-lex's afford a lower level of sister chromatid exchange (SCE) in 9L cells that are sensitive to CENU. There is no difference between CENU-lex in SCE induction in 9L-2 cells that are resistant to CENU. Formation of DNA interstrand crosslinks from the CENU-lex's is lower than for their nonaffinity binding analogs in low ionic strength buffer, but similar in the same buffer containing 200 mM NaCl. Salt inhibits crosslinking for all CENU, but distamycin, a competitive inhibitor of lex minor groove binding, uniquely enhances crosslinks for the CENU-lex's. These results are consistent with the novel minor groove adduction being a 'detoxification' pathway for the CENU-lex's since this lesion is formed at the expense of the cytotoxic major groove interstrand crosslink.