DNA bis-intercalators as new anti-tumour agents: modulation of the anti-tumour activity by the linking chain rigidity in the ditercalinium series.

Ditercalinium (2,2'-([4,4'-bipiperidine-1,1'-diyl] di-2,1-ethane diyl) bis (10-methoxy-7H-pyrido[4,3c] carbazolium) tetra(methyl sulphonate--NSC 366241), a DNA bis-intercalating compound presently under clinical trial, elicits an original mechanism of action and thus appears as the first of a new class of anti-tumour drugs. Previous studies have shown that a reduced flexibility of the linking chain of these dimers is essential for their biological activity. In order to analyze their mechanism of action at the molecular level and to obtain structure-activity relationships in this series, new derivatives with additional methylene groups between the two piperidine rings have been synthesized. Whereas the addition of a single methylene group in the chain preserves the anti-tumour activity of the dimers, the addition of a second methylene diminishes it; the addition of three methylenes completely abolishes it. Lengthening of sonicated DNA and unwinding of supercoiled DNA support a bis-intercalation mechanism for these drugs. In addition, analyses of poly d(A-T) melting curves in the presence of the drugs, and competition experiments with ethidium dimer, show that these compounds bind to DNA with high affinity (10(7)-10(8) M-1). N.m.r. studies of the dimers in aqueous medium show that the introduction of a single methylene group in the linker leads to compounds with a conformationally-induced decrease of intermolecular stacking interactions, which might be related to the DNA affinity enhancement observed for these dimers. Different hypotheses concerning structure-activity relationships in the different series are discussed.