68Ga labeled Olmutinib: Design, synthesis, and evaluation of a novel PET EGFR probe.

Abstract

Radiolabeled tyrosine kinase inhibitors (TKIs) offer a promising approach for molecular imaging of EGFR-positive cancers. Despite the development of various EGFR small-molecule probes, none of the ⁶⁸Ga-labeled small-molecule probes based on the chelator DOTA have shown tumor-specific uptake. To address this challenge, we selected Olmutinib, a third-generation EGFR covalent inhibitor, as a PET imaging tracer for EGFR-positive tumors. We synthesized the precursor DOTA-Olmutinib through a five-step process and subsequently radiolabeled it with ⁶⁸Ga to prepare ⁶⁸Ga-DOTA-Olmutinib. ⁶⁸Ga-DOTA-Olmutinib displayed moderate lipophilicity (log P = 0.85) and exhibited high stability in vitro and in vivo. Western blot analysis was used to detect the level of EGFR in multiple tumor cells. In cell uptake experiments, ⁶⁸Ga-DOTA-Olmutinib exhibited enhanced uptake specifically in tumor cells with a higher level of EGFR supporting it as an EGFR-specific tracer. Additionally, PET/CT imaging with ⁶⁸Ga-DOTA-Olmutinib showed significant tumor uptake at 60 min with 4 % ID/g post-injection, marking a breakthrough, though the uptake is not yet ideal. Overall, our results suggest that ⁶⁸Ga-labeled Olmutinib holds promise as a potential PET tracer for detecting EGFR-positive cancers.