68Ga-NOTA-m-SNA006: A Next-Generation CD8-Targeting Nanobody Probe for
Enhanced Renal and Hepatic Clearance in Noninvasive ImmunoPET Imaging
Abstract
Background and purpose The 68Ga-labeled nanobody SNA006 marks a
significant advancement in noninvasive ImmunoPET imaging of CD8+ T
cells, facilitating real-time tracking of cellular immune responses in
cancer, yet its pharmacokinetic properties remain suboptimal. This study
aimed to develop a next-generation CD8-targeting immunoPET nanobody
probe by incorporating a PEGylated brush border membrane
enzyme-cleavable linker to improve pharmacokinetics and to evaluate its
characterization in CD8-positive intrapulmonary tumors. Experimental
approach A precursor based on SNA006, containing a PEGylated brush
border membrane enzyme-cleavable linker, was designed, synthesized, and
radiolabeled with gallium-68 to yield 68Ga-NOTA-m-SNA006. The probe was
subsequently assessed both in vitro and in vivo. Key results The probe
exhibited high radiochemical yield, purity, and favorable stability, and
demonstrated binding to the CD8 protein with high affinity. PET/CT
imaging and biodistribution studies revealed that 68Ga-NOTA-m-SNA006
exhibited favorable pharmacokinetic properties, including rapid
clearance from the kidneys, reduced liver uptake, and sustained
retention in the tumor, compared with 68Ga-NODAGA-SNA006.
68Ga-NOTA-m-SNA006 exhibited high uptake in lung lesions during in vivo
PET imaging, reflecting CD8 expression in an intrapulmonary tumor model.
Conclusion and implication In summary, we present a novel 68Ga-labeled
SNA006 radiotracer with an optimized linker moiety, 68Ga-NOTA-m-SNA006,
which effectively decreases renal and hepatic uptake while maintaining
tumor uptake, thereby enhancing the tumor-to-background ratio. This
approach represents a critical advancement in addressing the persistent
challenge of radioactivity in the kidneys and liver associated with
nanobody-based radiopharmaceuticals.