Synthesis and In Vitro Comparison of DOTA, NODAGA and 15-5 Macrocycles as Chelators for the <sup>64</sup>Cu-Labelling of Immunoconjugates

oleh: Aurélie Maisonial-Besset, Tiffany Witkowski, Mercedes Quintana, Sophie Besse, Vincent Gaumet, Axel Cordonnier, Cyrille Alliot, Aurélien Vidal, Caroline Denevault-Sabourin, Sébastien Tarrit, Sophie Levesque, Elisabeth Miot-Noirault, Jean-Michel Chezal

Format: Article
Diterbitkan: MDPI AG 2022-12-01

Deskripsi

The development of <sup>64</sup>Cu-based immuno-PET radiotracers requires the use of copper-specific bifunctional chelators (BFCs) that contain functional groups allowing both convenient bioconjugation and stable copper complexes to limit in vivo bioreduction, transmetallation and/or transchelation. The excellent in vivo kinetic inertness of the pentaazamacrocyclic [<sup>64</sup>Cu]Cu-15-5 complex prompted us to investigate its potential for the <sup>64</sup>Cu-labelling of monoclonal antibodies (mAbs), compared with the well-known NODAGA and DOTA chelators. To this end, three NODAGA, DOTA and 15-5-derived BFCs, containing a pendant azadibenzocyclooctyne moiety, were synthesised and a robust methodology was determined to form covalent bonds between them and azide-functionalised trastuzumab, an anti-HER2 mAb, using strain-promoted azide-alkyne cycloaddition. Unlike the DOTA derivative, the NODAGA- and 15-5-mAb conjugates were radiolabelled with <sup>64</sup>Cu, obtaining excellent radiochemical yields, under mild conditions. Although all the radioimmunoconjugates showed excellent stability in PBS or mouse serum, [<sup>64</sup>Cu]Cu-15-5- and [<sup>64</sup>Cu]Cu-NODAGA-trastuzumab presented higher resistance to transchelation when challenged by EDTA. Finally, the immunoreactive fraction of the radioimmunoconjugates (88–94%) was determined in HER-2 positive BT474 human breast cancer cells, confirming that the bioconjugation and radiolabelling processes implemented had no significant impact on antigen recognition.