The radioprotective agent WR 2721 has been shown to prevent deterioration of renal function in a rodent model of chronic renal failure, and it has been suggested that WR 2721 may exert these protective effects by scavenging free radicals. To test this hypothesis, we investigated the effects of WR 2721 and its dephosphorylated, active metabolite, WR 1065, in an in vitro model of oxidative injury. By using the classical assay for superoxide production (reduction of ferricytochrome c), we first demonstrated that WR 1065 may act as a reducing agent. To establish the biological relevance of this reducing ability, U937 cells (a human premonocytic line) were incubated with or without WR 1065 or 2721, then exposed to increasing concentrations of H2O2. Cell viability was measured by Trypan blue exclusion and [3H]-thymidine incorporation into DNA, and cytosolic free calcium determined by fura-2 fluorescence. WR 1065 protected U937 cells from H2O2-induced cell death in a dose-dependent manner, and more efficiently than WR 2721. WR 1065 also prevented the rise in cytosolic free calcium induced in these cells by H2O2. These in vitro molecular and cellular events may account, at least in part, for the in vivo protective effects of WR 2721.