The 2,2'-bipyridyl-6-carbothioamide copper (II) complex differs from the iron (II) complex in its biochemical effects in tumor cells, suggesting possible differences in the mechanism leading to cytotoxicity.
2,2'-bipyridyl-6-carbothioamide (BPYTA) is an antitumor agent with chelating properties. It has previously been shown that the R2 subunit of ribonucleotide reductase (RR) is its major cellular target, but RR inhibition is observed only in the presence of ferrous iron (BPYTA-Fe, molar ratio 2:1). Because the copper (II) complex of BPYTA (BPYTA-Cu, molar ratio 1:1)) has in vitro antitumor activity comparable to that of BPYTA-Fe, we studied the mechanism of action of this new metal complex. Spectorphotometric and HPLC studies demonstrated that, at pH 7.5, BPYTA-Cu is stable at molar ratio 2:1 and copper is in its favored oxidized form [BPYTA-Cu(II)]. Electron paramagnetic resonance (EPR) studies with mouse recombinant R2 demonstrated that BPYTA-Cu destroys the R2 tyrosyl radical at the same concentration at which BPYTA-Fe does (78% vs 73% destruction at 200 microM, with 5 min of contact), but R2 inhibition is not time-dependent. Studies of the metabolism of [14C] cytidine suggest that the cytotoxic activity of BPYTA-Cu can be explained in terms of RR inhibition. However, the significant inhibition of RNA synthesis and the lack of cross-resistance to BPYTA-Cu of cell lines resistant to other RR inhibitors suggest that BPYTA-Cu may have more than one cellular target. Moreover, cell proliferation studies suggest that, unlike BPYTA-Fe, BPYTA-Cu displays its activity immediately after contact with the target cells. Our study demonstrates that significant differences in the biochemical effects of BPYTA and, perhaps, also its mechanism of action are due solely to the bonded transition metalloelement. This might also be the case with other chelators that demonstrate cytotoxic activity following metalloelement chelation.
Nocentini
G
G
Department of Clinical Medicine, Pathology and Pharmacology, University of Perugia, Italy.
Barzi
A
A
D015082:2,2'-Dipyridyl / Q000031:analogs & derivatives* / Q000494:pharmacology; D000818:Animals; D000970:Antineoplastic Agents / Q000378:metabolism / Q000494:pharmacology*; D005296:Ferrous Compounds / Q000494:pharmacology*; D051379:Mice; D009942:Organometallic Compounds / Q000494:pharmacology*; D012313:RNA / Q000235:genetics; D012264:Ribonucleotide Reductases / Q000037:antagonists & inhibitors / Q000235:genetics; D014407:Tumor Cells, Cultured
C099973:2,2'-bipyridyl-6-carbothioamide copper(II) complex; C078811:2,2'-bipyridyl-6-carbothioamide-ferrous complex; D000970:Antineoplastic Agents; D005296:Ferrous Compounds; D009942:Organometallic Compounds; D015082:2,2'-Dipyridyl; D012313:RNA; D012264:Ribonucleotide Reductases