Comparison of the octadentate bifunctional chelator DFO*-pPhe-NCS and the clinically used hexadentate bifunctional chelator DFO-pPhe-NCS for 89Zr-immuno-PET

All clinical Zr-89-immuno-PET studies are currently performed with the chelator desferrioxamine (DFO). This chelator provides hexadentate coordination to zirconium, leaving two coordination sites available for coordination with, e.g., water molecules, which are relatively labile ligands. The unsaturated coordination of DFO to zirconium has been suggested to result in impaired stability of the complex in vivo and consequently in unwanted bone uptake of Zr-89. Aiming at clinical improvements, we report here on a bifunctional isothiocyanate variant of the octadentate chelator DFO* and the in vitro and in vivo comparison of its Zr-89-DFO*-mAb complex with Zr-89-DFO-mAb. The bifunctional chelator DFO*-pPhe-NCS was prepared from previously reported DFO* and p-phenylenediisothiocyanate. Subsequently, trastuzumab was conjugated with either DFO*-pPhe-NCS or commercial DFO-pPhe-NCS and radiolabeled with Zr-89 according to published procedures. In vitro stability experiments were carried out in saline, a histidine/sucrose buffer, and blood serum. The in vivo performance of the chelators was compared in N87 tumor-bearing mice by biodistribution studies and PET imaging. In 0.9 % NaCl Zr-89-DFO*-trastuzumab was more stable than Zr-89-DFO-trastuzumab; after 72 h incubation at 2-8 A degrees C 95 % and 58 % intact tracer were left, respectively, while in a histidine-sucrose buffer no difference was observed, both products were ae 92 % intact. In vivo uptake at 144 h post injection (p.i.) in tumors, blood, and most normal organs was similar for both conjugates, except for skin, liver, spleen, ileum, and bone. Tumor uptake was 32.59 +/- 11.95 and 29.06 +/- 8.66 % ID/g for Zr-89-DFO*-trastuzumab and Zr-89-DFO-trastuzumab, respectively. The bone uptake was significantly lower for Zr-89-DFO*-trastuzumab compared to Zr-89-DFO-trastuzumab. At 144 h p.i. for Zr-89-DFO*-trastuzumab and Zr-89-DFO-trastuzumab, the uptake in sternum was 0.92 +/- 0.16 and 3.33 +/- 0.32 % ID/g, in femur 0.78 +/- 0.11 and 3.85, +/- 0.80 and in knee 1.38 +/- 0.23 and 8.20 +/- 2.94 % ID/g, respectively. The uptake in bone decreased from 24 h to 144 h p.i. about two fold for the DFO* conjugate, while it increased about two fold for the DFO conjugate. Zr-DFO*-trastuzumab showed superior in vitro stability and in vivo performance when compared to Zr-89-DFO-trastuzumab. This makes the new octadentate DFO* chelator a candidate successor of DFO for future clinical Zr-89-immuno-PET.