Energetics, conformation, and recognition of DNA duplexes containing a major adduct of an anticancer azolato-bridged dinuclear Pt(II) complex.
BACKGROUND
The design of anticancer metallodrugs is currently focused on platinum complexes which form on DNA major adducts that cannot readily be removed by DNA repair systems. Hence, antitumor azolato-bridged dinuclear Pt(II) complexes, such as [{cis-Pt(NH(3))(2)}(2)(μ-OH)(μ-pyrazolate)](2+) (AMPZ), have been designed and synthesized. These complexes exhibit markedly higher toxic effects in tumor cell lines than mononuclear conventional cisplatin.


METHODS
Biophysical and biochemical aspects of the alterations induced in short DNA duplexes uniquely and site-specifically modified by the major DNA adduct of AMPZ, namely 1,2-GG intrastrand cross-links, were examined. Attention was also paid to conformational distortions induced in DNA by the adducts of AMPZ and cisplatin, associated alterations in the thermodynamic stability of the duplexes, and recognition of these adducts by high-mobility-group (HMG) domain proteins.


RESULTS
Chemical probing of DNA conformation, DNA bending studies and translesion synthesis by DNA polymerase across the platinum adduct revealed that the distortion induced in DNA by the major adduct of AMPZ was significantly less pronounced than that induced by similar cross-links from cisplatin. Concomitantly, the cross-link from AMPZ reduced the thermodynamic stability of the modified duplex considerably less. In addition, HMGB1 protein recognizes major DNA adducts of AMPZ markedly less than those of cisplatin.


CONCLUSIONS
The experimental evidence demonstrates why the major DNA adducts of the new anticancer azolato-bridged dinuclear Pt(II) complexes are poor substrates for DNA repair observed in a previously published report. The relative resistance to DNA repair explains why these platinum complexes show major pharmacological advantages over cisplatin in tumor cells.
Department of Biophysics, Faculty of Science, Palacky University, 17. listopadu 12, CZ-77146 Olomouc, Czech Republic. sandrock@centrum.cz
Jarmila
J
Mlcouskova
Jaroslav
J
Malina
Vojtech
V
Novohradsky
Jana
J
Kasparkova
Seiji
S
Komeda
Viktor
V
Brabec
D000818:Animals; D000970:Antineoplastic Agents / Q000737:chemistry* / Q000378:metabolism* / Q000494:pharmacology; D001483:Base Sequence; D002152:Calorimetry, Differential Scanning; D002945:Cisplatin / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology; D018736:DNA Adducts / Q000737:chemistry* / Q000378:metabolism*; D004734:Energy Metabolism / Q000502:physiology*; D006801:Humans; D008954:Models, Biological; D008968:Molecular Conformation; D015335:Molecular Probes / Q000737:chemistry; D009690:Nucleic Acid Conformation; D009691:Nucleic Acid Denaturation / Q000187:drug effects; D009944:Organoplatinum Compounds / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology; D010984:Platinum / Q000737:chemistry* / Q000378:metabolism; D058105:Polymerization / Q000187:drug effects; D051381:Rats
C578704:((cis-Pt(NH(3))(2))(2)(mu-OH)(mu-pyrazolate))(2+); D000970:Antineoplastic Agents; D018736:DNA Adducts; D015335:Molecular Probes; D009944:Organoplatinum Compounds; D010984:Platinum; D002945:Cisplatin