Stability Estimation of Gallium Complexes of DOTA Derivatives for Radiotheranostic Applications

1,4,7,10-Tetraazacyclododecane-1,4,7-triacetic acid (DO3A) has been used to prepare ⁶⁸Ga-labeled probes for the diagnostic counterpart of radiotheranostic applications. While DO3A provides stable complexes with therapeutic radionuclides such as ⁹⁰Y, ¹⁷⁷Lu, and ²²⁵Ac, further improvement of the in vivo stability of the Ga-DO3A complex is required. Considering the high stability of an intact Ga-DOTA complex, the stability of Ga complexes of DOTA and DO3A derivatives, including benzyl-DOTA (Bn-DOTA), was evaluated to gain fundamental knowledge for developing the next-generation radiotheranostic probes using ⁶⁸Ga as a diagnostic counterpart. Following the complexation reaction to prepare ⁶⁷Ga-labeled DOTA and DO3A derivatives, the stability of the resulting ⁶⁷Ga-labeled compounds was evaluated in murine plasma and apo-transferrin challenge. [⁶⁷Ga]Ga-Bn-DOTA produced two isomers, and one of the isomers exhibited the highest stability among the tested complexes. The X-ray crystallography showed that the less stable isomer of Ga-Bn-DOTA suggested an N₃O₃ coordination geometry, while Ga-DOTA and Ga-Bn-DO3A show N₄O₂ coordination. To further evaluate the stability, a synthetic somatostatin analogue, [Tyr³]octreotide (TOC), was used as a model peptide, and p-COOH-Bn-DOTA and DO3A were conjugated with TOC to prepare DOTA-Bn-TOC and DOTATOC. [⁶⁷Ga]Ga-DOTA-Bn-TOC also yielded two isomers with varying stability, and one isomer exhibited significantly higher stability than [⁶⁷Ga]Ga-DOTATOC both in vitro and in vivo. These findings indicate that para-substituted Bn-DOTA would constitute a suitable chelating agent for developing next-generation radiotheranostic probes, although high-performance liquid chromatography purification is needed. Thus, further chemical modification on the Bn-DOTA molecule is also needed to avoid the formation of a Ga complex with the N₃O₃ configuration.