Using cell-fractionation and photochemical crosslinking methods to determine the cellular binding site(s) of the antitumor drug DMP 840.
In order to understand its mechanism of action we have begun an effort to better define the cellular target of action of the experimental antitumor agent DMP 840 (NSC D640430; (R,R)-2,2'-(1,2-ethanediylbis(imino-(1-methyl-2,1-ethanediyl)))-bi s(5- nitro-1H-benz(de)isoquinoline-1,3-(2H)-dione) dimethanesulfonate). Using a combination of gentle cell fractionation procedures and a previously unidentified photochemical crosslinking reaction, we have shown that after the drug is added to cultured Clone A cells, more than 80% of the drug that is found associated with cells partitions to the chromatin-containing structural framework of the cell and that the primary target after crosslinking with 360 nm light is DNA. While DMP 840 photoreacts quite efficiently with purified RNA in vitro, no photoattachment of the drug to RNA was observed in cells. In vitro photochemical studies also reveal that while GC-rich DNA is a preferred target for drug interaction, AT-rich DNA is more active in the photochemical crosslinking reaction. These results suggest that DMP 840 probably kills cells by interfering with DNA-metabolic processes, and that the drug and its derivatives are likely to be useful photoactive molecular probes for investigating higher order chromatin structures in cells.
Chatterjee
P K
PK
Cancer Research Group, DuPont Merck Pharmaceutical Company, Glenolden, PA 19036, USA.
Sternberg
N L
NL
D000818:Animals; D000970:Antineoplastic Agents / Q000378:metabolism* / Q000494:pharmacology / Q000528:radiation effects; D001665:Binding Sites; D002417:Cattle; D002458:Cell Fractionation; D003432:Cross-Linking Reagents; D004247:DNA / Q000378:metabolism; D006801:Humans; D066298:In Vitro Techniques; D007546:Isoquinolines / Q000378:metabolism* / Q000494:pharmacology / Q000528:radiation effects; D008698:Mesylates / Q000378:metabolism* / Q000494:pharmacology / Q000528:radiation effects; D010777:Photochemistry; D011119:Polynucleotides / Q000378:metabolism; D014407:Tumor Cells, Cultured
D000970:Antineoplastic Agents; D003432:Cross-Linking Reagents; D007546:Isoquinolines; D008698:Mesylates; D011119:Polynucleotides; C084899:bisnafide; D004247:DNA