Hypoxia-selective activation of 5-fluorodeoxyuridine prodrug possessing indolequinone structure: radiolytic reduction and cytotoxicity characteristics.

We synthesized a 5-fluorodeoxyuridine (5-FdUrd) derivative possessing an indolequinone structure (IQ-FdUrd) to characterize the radiolytic reduction in aqueous solution and the radiation-activated cytotoxicity against EMT6/KU cells under hypoxic conditions. IQ-FdUrd released antitumor agent 5-FdUrd upon hypoxic, but not aerobic, irradiation with the G value of 0.38 x 10(-7) mol J(-1). Laser flash photolysis of IQ-FdUrd in Ar-purged aqueous solution with dimethylaniline as an electron donor gave rise to a transient absorption spectrum characteristic of semiquinone radical anion, which decayed via second order kinetics. It is most likely that bimolecular disproportionation of intermediate semiquinone radicals occurs to release 5-FdUrd. IQ-FdUrd showed enhanced cytotoxicity against EMT6/KU cells in a radiation dose-dependent manner upon hypoxic irradiation. IQ-FdUrd is potentially a prototype compound for new class of radiation-activated antitumor prodrugs that are useful for radiation treatment of hypoxic tumors.

In vitro and in vivo evaluation of novel antitumor prodrugs of 5-fluoro-2'-deoxyuridine activated by hypoxic irradiation.

PURPOSE: We previously developed a novel antitumor prodrug that has a 2-oxopropyl substituent at the N(1) position of 5-fluorouracil (5-FU) and releases 5-FU via one-electron reduction on hypoxic irradiation. Although the compound was effective in vivo, its activity against murine tumors was not high enough to warrant clinical studies. Therefore, we developed a similar family of radiation-activated prodrugs of 5-fluoro-2'-deoxyuridine (FdUrd), which is generally more potent than 5-FU, and investigated their radiation chemical reactivity and in vitro and in vivo effects. METHODS AND MATERIALS: Compounds bearing various 2-oxoalkyl substituents at the N(3) position of FdUrd were synthesized and investigated. After aerobic or hypoxic irradiation to the prodrugs dissolved in water or culture medium, release of FdUrd was measured using high-performance liquid chromatography. To investigate in vitro cytotoxicity, SCCVII and EMT6 cells in culture were irradiated in the presence of the prodrug under aerobic or hypoxic conditions, and then kept with the compound for 24 h. Cell survival was then measured using a colony assay. To investigate in vivo effects, the drug was injected intraperitoneally at a dose of 100 or 300 mg/kg into Balb/c mice bearing EMT6 tumors 30 min before irradiation. The tumor growth delay-time was then assessed. RESULTS: In vitro, the prodrugs released FdUrd at G-values (molar numbers of molecules produced by 1 J of radiation energy) of 1.6-2.0 x 10(-7) mol/J after hypoxic irradiation. The G-values for FdUrd release with hypoxic irradiation were about 100-fold greater than those with aerobic irradiation. Among the prodrugs tested, OFU106 bearing a 2-oxocyclopentyl substituent released the highest amount of FdUrd in the culture medium, and it was subjected to further in vitro and in vivo assays. Although OFU106 administered alone showed no cytotoxicity up to a concentration of 0.2 mM, it produced an enhanced cytotoxic effect when administered before hypoxic irradiation and kept with the cells for 24 h. The enhancement ratios calculated at the surviving fraction of 1% were 1.35-1.4 at 0.04 mM and 1.45-1.5 at 0.2 mM. In vivo, however, administration of OFU106 (100 or 300 mg/kg) before 20 Gy of irradiation did not produce marked growth delays compared with 20 Gy of radiation alone. CONCLUSION: On hypoxic irradiation in vitro, the prodrugs of FdUrd were activated as efficiently as were the prodrugs of 5-FU, but marked in vivo effects could not be detected. This strategy of prodrug design should be used in further development of radiation-activated prodrugs of more potent anticancer agents.

One-electron reduction characteristics of N(3)-substituted 5-fluorodeoxyuridines synthesized as radiation-activated prodrugs.

We designed and synthesized N(3)-substituted 5-fluorodeoxyuridines as radiation-activated prodrugs of the antitumor agent, 5-fluorodeoxyuridine (5-FdUrd). A series of 5-FdUrd derivatives possessing a 2-oxoalkyl group at the N(3)-position released 5-FdUrd in good yield via one-electron reduction initiated by hypoxic irradiation. Cytotoxicity of the 5-FdUrd derivative possessing the 2-oxocyclopentyl group (3d) was low, but was enhanced by hypoxic irradiation resulting in 5-FdUrd release.