title: Radiolabeling and in vitro and in vivo evaluation of [18F]-FE-DABP688 as a PET radioligand for the metabotropic glutamate receptor subtype 5.
abstract: BACKGROUND
  Fluoroethyl-desmethyl-ABP688 (FE-DABP688) is a novel derivative of the previously described positron emission tomography (PET) ligand 3-(6-methyl-pyridin-2-ylethynyl)-cyclohex-2-enone-O-[11C]-methyl-oxime. FE-DABP688 was radiolabeled with fluorine-18 and characterized as a PET imaging agent for the metabotropic glutamate receptor subtype 5 (mGluR5).
  
  
  METHODS
  FE-DABP688 was radiolabeled by reacting 2-[18F]-fluoroethyl tosylate with the sodium salt of 3-(pyridin-2-ylethynyl)-cyclohex-2-enone-oxime in dry DMF. The in vitro affinity of [18F]-FE-DABP688 for mGluR5 was determined by Scatchard analysis of saturation binding data using rat whole-brain membranes (without cerebellum). Further in vitro characterization of the tracer involved plasma stability and lipophilicity testing. In vivo evaluation of [18F]-FE-DABP688 was performed by postmortem biodistribution experiments and PET studies in rats using the dedicated small-animal PET tomograph quad-HIDAC.
  
  
  RESULTS
  The radiotracer was obtained in good radiochemical yields in an overall synthesis time of 150 min. The radiochemical yield after semipreparative HPLC was 25+/-8% (n>7, decay corrected), and specific activity was 30+/-5 GBq/micromol (n>7). [18F]-FE-DABP688 exhibited optimal lipophilicity with a logD value of 2.1+/-0.1 and high plasma stability. Saturation assays of [(18)F]-FE-DABP688 revealed a single high-affinity binding site with a dissociation constant (Kd) of 1.6+/-0.4 nM and a Bmax value of 119+/-24 fmol/mg protein. PET scanning indicated radioactivity uptake in mGluR5-rich regions such as the hippocampus, striatum and cortex, while radioactivity accumulation in the cerebellum, a region with negligible mGluR5 density, was significantly lower. Biodistribution studies showed a similar distribution pattern of [18F]-FE-DABP688 binding in the brain. The hippocampus-to-cerebellum and striatum-to-cerebellum ratios were 1.81+/-0.16 and 1.93+/-0.36, respectively. Blocking studies using coinjection of [18F]-FE-DABP688 and unlabeled 2-methyl-6-((3-methoxyphenyl)ethynyl)-pyridine (1 mg/kg) revealed more than 45% specific binding in the hippocampus and striatum, thus demonstrating the in vivo specificity of tracer binding.
  
  
  CONCLUSIONS
  [18F]-FE-DABP688 may be a useful PET tracer for imaging mGluR5 in rodents.
authors.affiliation: Animal Imaging Center - PET, Center for Radiopharmaceutical Science of ETH, PSI and USZ, ETH Hoenggerberg, CH-8093 Zurich, Switzerland. michael.honer@pharma.ethz.ch
authors.forename: Michael
authors.initials: M
authors.lastname: Honer
authors.forename: Anja
authors.initials: A
authors.lastname: Stoffel
authors.forename: Lea J
authors.initials: LJ
authors.lastname: Kessler
authors.forename: P August
authors.initials: PA
authors.lastname: Schubiger
authors.forename: Simon M
authors.initials: SM
authors.lastname: Ametamey
mesh_terms: D000818:Animals; D001921:Brain / Q000000981:diagnostic imaging* / Q000378:metabolism*; D007553:Isotope Labeling / Q000379:methods; D008297:Male; D008657:Metabolic Clearance Rate; D009928:Organ Specificity; D010091:Oximes / Q000737:chemistry / Q000493:pharmacokinetics*; D049268:Positron-Emission Tomography / Q000379:methods*; D011725:Pyridines / Q000737:chemistry / Q000493:pharmacokinetics*; D019275:Radiopharmaceuticals / Q000138:chemical synthesis / Q000493:pharmacokinetics; D051381:Rats; D017207:Rats, Sprague-Dawley; D064529:Receptor, Metabotropic Glutamate 5; D018094:Receptors, Metabotropic Glutamate / Q000378:metabolism*; D014018:Tissue Distribution
chemical_list: C578937:Grm5 protein, rat; D010091:Oximes; D011725:Pyridines; D019275:Radiopharmaceuticals; D064529:Receptor, Metabotropic Glutamate 5; D018094:Receptors, Metabotropic Glutamate; C526997:fluoroethyl-desmethyl-ABP688