A combi-nitrosourea prodrug releasing DNA cross-linking agent and AGT inhibitor was synthesized.
The combi-nitrosourea exhibits higher cytotoxicity against mer+ glioma cells than ACNU.
The prodrug induces higher levels of dG–dC crosslink than traditional CENU chemotherapies.
AGT activity is efficiently inhibited by the O6-benzylguanine analog released from the prodrug.