Synthesis of [1-¹³ C-5-¹² C]-alpha-ketoglutarate enables noninvasive detection of 2-hydroxyglutarate

Isocitrate dehydrogenase 1 (IDH1) mutations that generate the oncometabolite 2-hydroxyglutarate (2-HG) from α-ketoglutarate (α-KG) have been identified in many types of tumors and are an important prognostic factor in gliomas. 2-HG production can be determined by hyperpolarized carbon-13 magnetic resonance spectroscopy (HP-¹³ C-MRS) using [1-¹³ C]-α-KG as a probe, but peak contamination from naturally occurring [5-¹³ C]-α-KG overlaps with the [1-¹³ C]-2-HG peak. Via a newly developed oxidative-Stetter reaction, [1-¹³ C-5-¹² C]-α-KG was synthesized. α-KG metabolism was measured via HP-¹³ C-MRS using [1-¹³ C-5-¹² C]-α-KG as a probe. [1-¹³ C-5-¹² C]-α-KG was synthesized in high yields, and successfully eliminated the signal from C5 of α-KG in the HP-¹³ C-MRS spectra. In HCT116 IDH1 R132H cells, [1-¹³ C-5-¹² C]-α-KG allowed for unimpeded detection of [1-¹³ C]-2-HG. ¹² C-enrichment represents a novel method to circumvent spectral overlap, and [1-¹³ C-5-¹² C]-α-KG shows promise as a probe to study IDH1 mutant tumors and α-KG metabolism.