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Trapping of DNA-reactive Metabolites of Therapeutic or Carcinogenic Agents by Carbon-14-labeled Synthetic Polynucleotides¹

Division of Oncology, Department of Medicine, Neil Hellman Medical Research Building, Albany Medical College, Albany, New York 12208

Many substances which do not react with DNA directly are metabolized into important DNA-modifying intermediates. We have devised a method for trapping these intermediates with ¹⁴C-labeled nucleosides contained in a synthetic polynucleotide. The polynucleotide structure protects the labeled nucleoside from metabolism; thus, it is unaltered when the polymer is incubated with a drug-metabolizing system. However, when the polymer is incubated with this system and a compound which can be metabolized into a reactive species, these intermediates are trapped by the ¹⁴C-labeled nucleoside and subsequently are detected as new peaks of radioactivity in a digest of the labeled polynucleotide. This system has been used to detect reactive intermediates of cyclophosphamide generated by a liver homogenate.

¹ Supported by Grants CA 20292 from the National Cancer Institute and CH 188 from the American Cancer Society.

² To whom requests for reprints should be addressed.

Received 4/15/81. Accepted 4/21/82.