Evaluation of 3-hydroxypyridin-2-one (2,3-HOPO) based macrocyclic chelator for 89Zr4+ and its use for immunoPET imaging of HER2 positive model of ovarian carcinoma in mice.

Objectives A novel octadentate 3-hydroxypyridin-2-one (2,3-HOPO) based di-macrocyclic ligand was evaluated for chelation of ⁸⁹Zr; subsequently, it was used as a bi-functional chelator for preparation of ⁸⁹Zr-labeled antibodies.

Methods Quantitative chelation of ⁸⁹Zr⁴⁺ with the octadentate ligand forming ⁸⁹ZrL complex was achieved under mild conditions within 15 minutes. The ⁸⁹Zr-complex was stable in vitro in presence of DTPA, but a slow degradation was observed in serum. In vivo, the hydrophilic ⁸⁹Zr-complex showed prevalently renal excretion; and an elevated bone uptake of radioactivity suggested a partial release of ⁸⁹Zr⁴⁺ from the complex.

Results The 2,3-HOPO based ligand was conjugated to the monoclonal antibodies, HER2-specific trastuzumab and an isotypic anti-gD antibody, using a p-phenylene bis-isothiocyanate linker to yield products with an average loading of less than 2 chelates per antibody. Conjugated antibodies were labeled with ⁸⁹Zr under mild conditions providing the PET tracers in 60-69% yield. Despite the limited stability in mouse serum; the PET tracers performed very well in vivo. The PET imaging in mouse model of HER2 positive ovarian carcinoma showed tumor uptake of ⁸⁹Zr-trastuzumab (29.2 ± 12.9 %ID/g) indistinguishable (p = 0.488) from the uptake of positive control ⁸⁹Zr-DFO-trastuzumab (26.1 ± 3.3 %ID/g).

Conclusions In conclusion, the newly developed 3-hydroxypyridin-2-one based di-macrocyclic chelator provides a viable alternative to DFO-based heterobifunctional ligands for preparation of ⁸⁹Zr-labeled monoclonal antibodies for immunoPET studies.