Design and Preclinical Evaluation of an Albumin-Binding PSMA Ligand for ⁶⁴Cu-Based PET Imaging

Recently, we developed an albumin-binding radioligand (177Lu–PSMA–ALB-56), which showed higher PSMA-specific tumor uptake in mice than the previously developed 177Lu–PSMA-617 under the same experimental conditions. Such a radioligand may be of interest also for PET imaging, possibly enabling better visualization of even small metastases at late time-points after injection. The aim of this study was, therefore, to modify PSMA–ALB-56 by exchanging the DOTA chelator with a NODAGA chelator for stable coordination of 64Cu (T1/2 = 12.7 h; Eβ+av = 278 keV). The resulting NODAGA-functionalized PSMA−ALB-89 ligand, and the previously establish DOTA-functionalized PSMA–ALB-56 ligand were labeled with 64Cu and evaluated in vitro and in vivo. Both radioligands showed plasma protein-binding properties in vitro and PSMA-specific uptake in PC-3 PIP cells. Biodistribution studies, performed in tumor-bearing mice, revealed high accumulation of 64Cu–PSMA–ALB-89 in PSMA-positive PC-3 PIP tumor xenografts (25.9 ± 3.41% IA/g at 1 h p.i.), which was further increased at later time-points (65.1 ± 7.82% IA/g at 4 h p.i. and 97.1 ± 7.01% IA/g at 24 h p.i.). High uptake of 64Cu–PSMA–ALB-89 was also seen in the kidneys, however, 64Cu–PSMA–ALB-89 was efficiently excreted over time. Mice injected with 64Cu–PSMA–ALB-56 showed increased accumulation of radioactivity in the liver (25.3 ± 4.20% IA/g) when compared to the liver uptake of 64Cu–PSMA–ALB-89 (4.88 ± 0.21% IA/g, at 4 h p.i.). This was most probably due to in vivo instability of the 64Cu–DOTA complex, which was also the reason for lower tumor uptake (49.7 ± 16.1% IA/g at 4 h p.i. and 28.3 ± 3.59% IA/g at 24 h p.i.). PET/CT imaging studies confirmed these findings and enabled excellent visualization of the PSMA-positive tumor xenografts in vivo after injection of 64Cu–PSMA–ALB-89. These data indicate that 64Cu–PSMA–ALB-89 is favorable over 64Cu–PSMA–ALB-56 with regard to the in vivo stability and tissue distribution profile. Moreover, 64Cu–PSMA–ALB-89 outperformed previously developed 64Cu-labeled PSMA ligands. Further optimization of long-circulating PSMA-targeting PET radioligands will be necessary before translating this concept to the clinics