Synthesis, radiolabeling, in vitro and in vivo evaluation of [18F]-FPECMO as a positron emission tomography radioligand for imaging the metabotropic glutamate receptor subtype 5

Nucl Med Biol. 2009 Aug;36(6):613-22. doi: 10.1016/j.nucmedbio.2009.03.005.

Abstract

Introduction: [18F]-(E)-3-((6-Fluoropyridin-2-yl)ethynyl)cyclohex-2-enone O-methyl oxime ([18F]-FPECMO) is a novel derivative of [11C]-ABP688. [18F]-FPECMO was characterized as a PET imaging agent for the metabotropic glutamate receptor subtype 5 (mGluR5).

Methods: [18F]-FPECMO was synthesized in a one-step reaction sequence by reacting [(18)F]-KF-K(222) complex with (E)-3-((6-bromopyridin-2-yl)ethynyl)cyclohex-2-enone O-methyl oxime in dry DMSO. The in vitro affinity of FPECMO was determined by displacement assays using rat whole brain homogenates (without cerebellum) and the mGluR5-specific radioligand [(3)H]-M-MPEP. Further in vitro characterization involved metabolite studies, lipophilicity determination and autoradiographical analyses of brain slices. In vivo evaluation was performed by postmortem biodistribution studies and PET experiments using Sprague-Dawley rats.

Results: The radiochemical yield after semipreparative HPLC was 35+/-7% and specific activity was >240 GBq/micromol. [(18)F]-FPECMO exhibited optimal lipophilicity (logD=2.1) and high metabolic stability in vitro. Displacement studies revealed a K(i) value of 3.6+/-0.7 nM for FPECMO. Biodistribution studies and ex vivo autoradiography showed highest radioactivity accumulation in mGluR5-rich brain regions such as the striatum and hippocampus. Co-injection of [18F]-FPECMO and ABP688 (1 mg/kg body weight), an mGluR5 antagonist, showed 40% specific binding in the striatum, hippocampus and cortex, regions known to contain high densities of the mGluR5. PET imaging, however, did not allow the visualization of mGluR5-rich brain regions in the rat brain due to a fast washout of [18F]-FPECMO from mGluR5-expressing tissues and rapid defluorination.

Conclusions: [18F]-FPECMO showed significant potential for the detection of mGluR5 in vitro; however, its in vivo characteristics are not optimal for a clear-cut visualization of the mGluR5 in rats.

MeSH terms

  • Animals
  • Autoradiography
  • Humans
  • Hydrophobic and Hydrophilic Interactions
  • Isotope Labeling
  • Ligands
  • Male
  • Oximes / chemical synthesis
  • Oximes / chemistry*
  • Oximes / metabolism
  • Positron-Emission Tomography
  • Pyridines / chemical synthesis
  • Pyridines / chemistry*
  • Pyridines / metabolism
  • Rats
  • Receptor, Metabotropic Glutamate 5
  • Receptors, Metabotropic Glutamate / metabolism*
  • Tissue Distribution

Substances

  • 3-((6-Fluoropyridin-2-yl)ethynyl)cyclohex-2-enone O-methyl oxime
  • 3-(6-methylpyridin-2-ylethynyl)cyclohex-2-enone-O-methyloxime
  • GRM5 protein, human
  • Grm5 protein, rat
  • Ligands
  • Oximes
  • Pyridines
  • Receptor, Metabotropic Glutamate 5
  • Receptors, Metabotropic Glutamate