Mechlorethamine-induced enhancement of radiation sensitivity of guanine.

Abstract

This study describes and characterizes the interactions of nitrogen mustard mechlorethamine (HN2) with guanine and the radiation sensitivity of guanine in the presence of HN2. Briefly, in an equimolar solution (0.5 mmol dm-3) the pH-dependence (pH 3.0-12.0) and time-dependence (0-36 h) of alkylation of guanine at room temperature were determined using a reverse-phase high-performance liquid chromatography (hplc) column. Based on the hplc peak areas of the product and intact guanine, the optimal pH for alkylation was determined to be 8.0. Similarly, the optimal time required for alkylation was 10 h. Two products, i.e. alkylated guanines, were detected (10:1, peak areas measured at 260 nm) and purified. Structural studies of the products were performed by direct insertion probe-electron impact mass spectrometry. These products were identified as N-(2-chloroethyl)-N-[2-(7-guanyl)ethyl]-methylamine (product 2). At optimal conditions, samples of either guanine or an equimolar solution of guanine and HN2 were 60Co irradiated (gamma-ray) at 25 Gy min-1 at doses up to 400 Gy. Both sets of samples were analysed by hplc. In each case, the sole radiation product observed and characterized was 8-hydroxy-guanine. Dose-yield plots were linear and showed that HN2 enhanced the radiation sensitivity of guanine. This increase in radiation sensitivity is attributed to the differences in electrophilic properties between nitrogen mustard and guanine.