Synthesis and evaluation of bifunctional DFO2K: a modular chelator with ideal properties for zirconium-89 chelation

Abstract

The synthesis and evaluation of the newest generation of our DFO2 chelator family—DFO2K—is described. DFO2K was designed with a simple synthetic route to access different bifunctional derivatives, with each derivative having similar metal ion coordination spheres and high denticity (up to 12 coordinate) to ensure stable coordination of zirconium-89. The high denticity could potentially enhance stability with other large oxophilic radiometals. Zirconium-89 is the most popular radionuclide to pair with large macromolecules such as antibodies (immunoPET) for positron emission tomography applications. Although clinically successful, the stability of the “gold standard” chelator desferrioxamine B (DFO) can be improved as significant bone uptake is observed in animal models, despite no obvious stability issues in humans. Following the synthesis of DFO2K we assessed its radiolabeling efficiency with zirconium-89 and compared with DFO, which revealed rapid and nearly identical radiolabeling kinetics to DFO. The resultant [⁸⁹Zr]Zr–DFO2K complex showed improved stability over [⁸⁹Zr]Zr–DFO in different in vitro stability assays such as hydroxyapatite and 1000-fold molar excess EDTA challenges. Furthermore, biodistribution studies of the non-bifunctional chelators in healthy mice showed that [⁸⁹Zr]Zr–DFO2K had a similar distribution profile and clearance to [⁸⁹Zr]Zr–DFO. The bifunctional derivative p-SCN–Ph–DFO2K was conjugated to a non-specific human IgG antibody and evaluated after 2 weeks circulating in healthy female CD1 mice. Mice administered [⁸⁹Zr]Zr–DFO2K–IgG showed substantially lower bone uptake in PET-CT images than [⁸⁹Zr]Zr–DFO–IgG, with PET ROI data and ex vivo biodistribution revealing a statistically significantly lower bone uptake for DFO2K. Overall, owing to its high denticity, ease of synthesis, improved solubility over DFO2 and DFO2p, and stable chelation of zirconium-89, DFO2K appears to be an improved alternative chelator to DFO for zirconium-89 chelation.