The interaction between the immune checkpoint PD-1 and PD-L1 promotes T-cell deactivation and cancer proliferation. Therefore, immune checkpoint inhibition therapy, which relies on prior assessment of the target, has been widely used for many cancers. As a non-invasive molecular imaging tool, radiotracers bring novel information on the in vivo expression of biomarkers (e. g., PD-L1), enabling a personalized treatment of patients. Our work aimed at the development of a PD-L1-specific, peptide-based PET radiotracer. We synthesized and evaluated a radiolabeled macrocyclic peptide adapted from a patent by Bristol Myers Squibb. Synthesis of [68 Ga]Ga-NJMP1 yielded a product with a radiochemical purity>95 % that was evaluated in vitro. However, experiments on CHO-K1 hPD-L1 cells showed very low cell binding and internalization rates of [68 Ga]Ga-NJMP1 in comparison to a control radiopeptide (WL12). Non-radioactive cellular assays using time-resolved fluorescence energy transfer confirmed the low affinity of the reported parent peptide and the DOTA-derivatives towards PD-L1. The results of our studies indicate that the macrocyclic peptide scaffold reported in the patent literature is not suitable for radiotracer development due to insufficient affinity towards PD-L1 and that C-terminal modifications of the macrocyclic peptide interfere with important ligand/receptor interactions.
Keywords: PD-1/PD−L1; PET; immune checkpoint blockade; peptides; radiopharmaceuticals.
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