Small Molecule Radiotracers for PET Imaging of PD-L1 with Copper-64

The programmed cell death ligand (PD-L1) is expressed on a number of different tumor entities and inhibits the immune response through binding to PD-1 on T-cells. Immune checkpoint inhibitors (ICI) prevent this blockade and thus can reactivate an immune response. However, only about 30% of the patients respond to an ICI monotherapy. Therefore, clinicians are in need for a non-invasive PET/SPECT radioligand for patient stratification and therapy monitoring.

Based on the structures of non-peptidic PD-L1 inhibitors, six different radiotracers were synthesized and radiolabelled with [64Cu]Cu2+ (HZDR, 30 MeV TR-FLEX cyclotron). Binding affinities were determined on PC3 cells stably overexpressing hPD-L1. For in vivo studies, qualitative PET/CT imaging experiments (nanoSCAN PET/CT, Mediso) were performed in NMRI-FoxN1-nude mice bearing PC3-hPD-L1 and mock xenograted tumors.

Two PD-L1 inhibitors were modified with strongly water-soluble acid groups, hydrophilic linker units and a NODAGA-chelator resulting in six different radioligands. The log(D) values of the copper-64 labelled radiotracers were between –3.17 and –4.15 and binding affinities ranged between 80.5 and 533 nM. Depending on the number and the pattern of sulfonate and phosphonate groups, in vivo experiments showed drastically different pharmacokinetic profiles. The radiotracer with one sulfonate and phosphonate group and the most hydrophobic linker exhibited a short circulation time, renal clearance, good tumor uptake (SUVmax = 3.5) and a distinct contrast between the hPD-L1 and the mock tumor.

In conclusion one PD-L1 radiotracer showed a promising pharmacokinetic profile, which is currently further modified to improve the binding affinity and tumor uptake.