NC-182 is a novel anti-tumour compound having a benzo[a]phenazine ring. Fluorescence, absorption and c.d. spectroscopy, as well as viscometric titrations, were systematically performed to investigate the interaction mode of this drug with DNA and its effect on DNA conformation, based on comparative measurements with distamycin (DNA minor-groove binder) and daunomycin (DNA-base intercalator). NC-182 was found to be a potent intercalator of DNA, especially the B-form DNA, although no specificity was observed against the base-pair. The binding of NC-182 to B-DNA behaves biphasically, depending on the molar ratio (r) of drug to DNA: NC-182 acts to render the B-form structure rigid at relatively low r value and to promote the transformation of B- to non-B forms at high r values. It was also shown that NC-182 promotes the unwinding of Z-form DNA to B-form. Viscometric, u.v. 'melting' and c.d. experiments further showed that (1) the DNA duplex structure is thermally stabilized by intercalation with NC-182 and (2) the intercalation of NC-182 into a poly(dA).2poly(dT) DNA structure thermally stabilizes the triplex structure, resulting in a melting point close to that of the duplex structure; the melting curves of triplex and duplex structures coincide at r > 0.06. These observations make a significant contribution to our understanding of the biological properties of this novel benzo[a]phenazine derivative, a new anti-tumour tumour agent against multidrug-resistant and sensitive tumours.