Synthesis of N-acetyl-l-aspartyl-l-glutamic acid; [3H]NAAG

³H]NAAG, N-acetyl-l -aspartyl-l -glutamic acid, has been widely used as a substrate in glutamate carboxypeptidase II (GCPII) reactions, either to determine the inhibitory constants at 50% inhibition (IC₅₀) of novel compounds or to measure GCPII activities in different tissues harvested from various disease models. The importance of ³H]NAAG, combined with its current commercial unavailability, prompted the development of a reliable eight-step synthetic procedure towards ³H₂]NAAG starting from commercially available pyroglutamate. Pure ³H]NAAG of high molar activity (49.8 Ci/mmol) and desired stereochemistry was isolated in high radiochemical yield (67 mCi) and radiochemical purity (>99%). The identity was confirmed by mass spectrometry and co-injection with unlabeled reference.