Characteristic Evaluation of a C-Labeled Leucine Analog, l-α-[5-C]methylleucine, as a Tracer for Brain Tumor Imaging by Positron Emission Tomography.

Amino acid transporters are upregulated in many cancer cells, and system L amino acid transporters (LAT1-4), in particular, LAT1, which preferentially transports large, neutral, and branched side-chain amino acids, are considered a primary target for cancer positron emission tomography (PET) tracer development. Recently, we developed a C-labeled leucine analog, l-α-[5-C]methylleucine ([5-C]MeLeu), via a continuous two-step reaction of Pd-mediated C-methylation and microfluidic hydrogenation. In this study, we evaluated the characteristics of [5-C]MeLeu and also compared the sensitivity to brain tumors and inflammation with l-[C]methionine ([C]Met) to determine its potential for brain tumor imaging. Competitive inhibition experiments, protein incorporation, and cytotoxicity experiments of [5-C]MeLeu were performed in vitro. Further, metabolic analyses of [5-C]MeLeu were performed using a thin-layer chromatogram. The accumulation of [5-C]MeLeu in tumor and inflamed regions of the brain was compared with [C]Met and C-labeled ()-ketoprofen methyl ester by PET imaging, respectively. Transporter assay with various inhibitors revealed that [5-C]MeLeu is mainly transported via system L amino acid transporters, especially LAT1, into A431 cells. The protein incorporation assay and metabolic assay in vivo demonstrated that [5-C]MeLeu was neither used for protein synthesis nor metabolized. These results indicate that MeLeu is very stable in vivo. Furthermore, the treatment of A431 cells with various concentrations of MeLeu did not change their viability, even at high concentrations (∼10 mM). In brain tumors, the tumor-to-normal ratio of [5-C]MeLeu was more elevated than that of [C]Met. However, the accumulation levels of [5-C]MeLeu were lower than those of [C]Met (the standardized uptake value (SUV) of [5-C]MeLeu and [C]Met was 0.48 ± 0.08 and 0.63 ± 0.06, respectively). In brain inflammation, no significant accumulation of [5-C]MeLeu was observed at the inflamed brain area. These data suggested that [5-C]MeLeu was identified as a stable and safe agent for PET tracers and could help detect brain tumors, which overexpress the LAT1 transporter.