Development of (68)Ga-labeled multivalent nitroimidazole derivatives for hypoxia imaging

Radiolabeled nitroimidazole (NI) derivatives have been extensively studied for imaging hypoxia. To increase the hypoxic tissue uptake, we developed (68)Ga-labeled agents based on mono-, bis-, and trisnitroimidazole conjugates with the chelating agent 1,4,7-triazacyclononane-1,4,7-tris[methyl(2-carboxyethyl)phosphinic acid] (TRAP). All the three agents showed high radiolabeling yields (>96%) and were found to be stable up to 4h in prepared medium at room temperature and in human serum at 37°C. The trivalent agent showed a significant increase in hypoxic to normoxic uptake ratio (p <0.005) according to the in vitro cell uptake experiments. Immunohistochemical analysis confirmed the presence of hypoxia in xenografted CT26 tumor tissue. The trivalent derivative ((68)Ga-3: 0.17±0.04, (68)Ga-4: 0.33±0.04, (68)Ga-5: 0.45±0.09, and (68)Ga-6: 0.47±0.05% ID/g) showed the highest uptake by tumor cells according to the biodistribution studies in CT-26 xenografted mice. All the nitroimidazole derivatives showed significantly higher uptake by tumor cells than the control agent (p <0.05) at 1h post-injection. The trivalent derivative ((68)Ga-3: 0.10±0.06; (68)Ga-4: 0.20±0.06; (68)Ga-5: 0.33±0.08; (68)Ga-6: 0.59±0.09) also showed the highest standard uptake value for tumor cells at 1h post-injection in animal PET studies using CT-26 xenografted mice. In conclusion, we successfully synthesized multivalent (68)Ga-labeled NI derivatives for imaging hypoxia. Among them, the trivalent agent showed the highest tumor uptake in biodistribution and animal PET studies.