Inactivation of O 6 ‐methylguanine‐DNA methyltransferase by glucose‐conjugated inhibitors

Abstract The DNA‐repair protein O 6 ‐methylguanine‐DNA methyltransferase (MGMT) is a decisive determinant of resistance of tumor cells to methylating and chloroethylating anti‐cancer drugs. Therefore, selective inhibition of MGMT in tumors is expected to cause tumor sensitization. Several inhibitors of MGMT have been developed which function in both tumors and normal tissue. To deplete MGMT preferentially in tumors, strategies to target the inhibitor to the tumor tissue need to be developed. Here, we report on the properties of glucose‐conjugated MGMT inhibitors that might be useful for tumor targeting since tumor cells frequently over‐express glucose transporter. O 6 ‐Benzylguanine (O6BG), 8‐aza‐ O 6 ‐benzylguanine, O 6 ‐(4‐bromothenyl)‐guanine (O6BTG) and the corresponding spacer‐linked β‐ D ‐glucose conjugates were analyzed comparatively for MGMT‐inhibitory activity. Substitution at the N9 position of the purine moiety resulted generally in a reduction in the efficiency with which the inhibitors blocked MGMT. However, the inhibitory activity of the O6BTG conjugates increased with increasing spacer length, and O6BTG conjugated with a C8 spacer with β‐ D ‐glucose was nearly as effective as O6BTG on its own. MGMT was inhibited by the conjugates both in crude cell extracts and upon treatment of intact HeLa cells, indicating efficient uptake of the glucose conjugates into cells. Since the O6BTG‐C8‐ D ‐glucose conjugate 8‐[ O 6 ‐(4‐bromothenyl)‐guan‐9‐yl]‐octyl‐β‐ D ‐glucoside was highly efficient at MGMT inhibition in a non‐toxic concentration range, the drug might be a useful tool for specific tumor sensitization. © 2001 Wiley‐Liss, Inc.