Synthesis and preclinical evaluation of two osimertinib isotopologues labeled with carbon-11 as PET tracers targeting the tyrosine kinase domain of the epidermal growth factor receptor.

Introduction: Osimertinib is a third-generation tyrosine kinase inhibitor (TKI) that is able to inhibit the EGFR treatment resistance mutation T790M and primary EGFR mutations Del19 and L858R. The aim of the study was to evaluate the potential of carbon-11 labeled osimertinib to be used as a tracer for the PET imaging of tumors bearing the T790M mutation.

Methods: Osimertinib was labeled with carbon-11 at two positions, and the effect of the labeling position on the metabolism and biodistribution was studied in female nu/nu mice. The mutation status specificity of osimertinib was confirmed in vitro in a cell growth inhibition experiment, and the tumor-targeting potential of the carbon-11 isotopologues was evaluated using female nu/nu mice xenografted with NSCLC cell lines; the wild-type EGFR expressing A549, the primary Del19 EGFR mutated HCC827 and the resistance T790M/L858R mutated H1975. One of the osimertinib tracers was selected based on the results acquired and evaluated for tracer specificity and selectivity by assessment of tumor uptake in a PET study where HCC827 tumor-bearing mice were pretreated with osimertinib or afatinib.

Results: [Methylindole-C]- and [dimethylamine-C]osimertinib were synthesized by C-methylation of precursors AZ5104 and AZ7550, respectively. Rapid metabolism of both analogs of [C]osimertinib was observed. Although the tumor uptake and retention of [methylindole-C]- and [dimethylamine-C]osimertinib in tumors were similar, the tumor-to-muscle ratios appeared to be higher for [methylindole-C]osimertinib. The highest uptake, tumor-to-blood, and tumor-to-muscle ratio were observed in the Del19 EGFR mutated HCC827 tumors. However, the specificity and selectivity of [methylindole-C]osimertinib PET could not be demonstrated in HCC827 tumors. The uptake of [methylindole-C]osimertinib was not significantly higher in T790M resistance mutated H1975 xenografts compared to the negative control cell line A549.

Conclusions: Osimertinib was successfully labeled at two positions with carbon-11, yielding two EGFR PET tracers, [methylindole-C]osimertinib and [dimethylamine-C]osimertinib. The preclinical evaluation demonstrated uptake and retention in three NSCLC xenografts; A549, HCC827, and H1975. The highest uptake was observed in the primary Del19 EGFR mutated HCC827. The ability of [methylindole-C]osimertinib to distinguish between the T790M resistance mutated H1975 xenografts and the wild-type EGFR expressing A549 could not be confirmed in the ex vivo study.