Radiolabelled CCK₂ R Antagonists Containing PEG Linkers: Design, Synthesis and Evaluation

The cholecystokinin-2/gastrin receptor (CCK₂ R) is considered a suitable target for the development of radiolabelled antagonists, due to its overexpression in various tumours, but no such compounds are available in clinical use. Therefore, we designed novel 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid-conjugated ligands based on CCK₂ R antagonist Z360/nastorazepide. As a proof of concept that CCK₂ R antagonistic activity can be retained by extending the Z360/nastorazepide structure using suitable linker, we present herein three compounds containing various PEG linkers synthesised on solid phase and in solution. The antagonistic properties were measured in a functional assay in the A431-CCK₂ R cell line (in the presence of agonist G17), with IC₅₀ values of 3.31, 4.11 and 10.4 nM for compounds containing PEG₄ , PEG₆ and PEG₁₂ , respectively. All compounds were successfully radiolabelled with indium-111, lutetium-177 and gallium-68 (incorporation of radiometal >95 %). The gallium-68-labelled compounds were stable for up to 2 h (PBS, 37 °C). log D_7.4 values were determined for indium-111- and gallium-68-labelled compounds, showing improved hydrophilicity compared to the reference compound.