Recent data suggest that expression of the membrane P170-glycoprotein (P-gp) may confer resistance to the
topoisomerase-I-interactive agent
topotecan. The present study describes the cellular effects of a new
dihydropyridine analogue,
PAK-200S, on P-gp-mediated resistance to
topotecan in human breast and ovarian tumour cells.
PAK-200S at a non-cytotoxic concentration of 2.0 microM completely reversed resistance to
topotecan in P-gp-expressing MCF-7/adr (breast) and A2780/Dx5 (ovarian) tumour cells, respectively, with no effects on parental cells. Cellular pharmacokinetic studies by reversed-phase high-performance liquid chromatography analysis showed significantly lower cellular
drug concentrations of the pharmacologically active closed-ring
lactone of
topotecan in multidrug-resistant cells than in parental cells.
PAK-200S was effective in restoring the cellular
lactone concentrations of
topotecan in resistant MCF-7/adr cells to levels comparable to those obtained in parental cells. Furthermore, exposure of MCF-7/adr cells to
topotecan in the presence of
PAK-200S significantly increased the induction of
protein-linked DNA breaks.
PAK-200S did not alter nuclear
topoisomerase I-mediated ex vivo pBR322
DNA plasmid unwinding activity and
topoisomerase-I protein expression. These results suggest that reversal of P-gp-mediated resistance to
topotecan by
PAK-200S was related to the restoration of cellular
drug concentrations of the active
lactone form of
topotecan rather than a direct effect on
topoisomerase-I function.