Assessment of DNA binding of platinum-radiosensitizer complexes by inhibition of restriction enzymes.
A simple and rapid method has been used to compare the binding of platinum complexes to DNA, in a relatively qualitative manner. A compound bound at or near the restriction site inhibits enzymatic cleavage of DNA; inhibition of BamHI and EcoRI activity by complexes was assessed in this study using linearized pSV2-gpt plasmid. Our particular interest was in DNA binding by complexes of platinum (Pt) with known organic radiosensitizers (RS), to determine whether the Pt was able to target the RS to the DNA. Although the Pt-RS complexes investigated themselves have moderate radiosensitizing ability (like the inorganic complexes, cis- or trans-diamminedichloroplatinum(II), c- or t-DDP) none of the Pt-RS inhibit to the same extent as c- or t-DDP. However, there appears to be some correlation between enhanced radiosensitization by Pt-RS over Pt(RS)2, with the degree of Pt binding (as assessed by our assay). Our results using isolated DNA suggest that not all complexes bind well (e.g. Pt with two RS ligands), but that in certain cases (e.g. Pt with only one RS), it is possible to target the drug to the DNA. An ammine or amine ligand may be required in order to target a radiosensitizer to DNA using platinum.
D001483:Base Sequence; D004247:DNA / Q000378:metabolism*; D004249:DNA Damage; D004262:DNA Restriction Enzymes; D009944:Organoplatinum Compounds / Q000378:metabolism*; D011838:Radiation-Sensitizing Agents / Q000378:metabolism*; D013054:Spectrophotometry, Atomic; D013329:Structure-Activity Relationship
D009944:Organoplatinum Compounds; D011838:Radiation-Sensitizing Agents; D004247:DNA; D004262:DNA Restriction Enzymes