title: 18F-FNDP for PET Imaging of Soluble Epoxide Hydrolase.
abstract: Soluble epoxide hydrolase (sEH) is a bifunctional enzyme located within cytosol and peroxisomes that converts epoxides to the corresponding diols and hydrolyzes phosphate monoesters. It serves to inactivate epoxyeicosatrienoic acids (EETs), which are generated in the brain to couple neuronal activity and cerebral blood flow in normal and pathologic states. Altered regulation of sEH was observed previously in various neuropathologic disorders including vascular dementia and stroke. Inhibitors of sEH are pursued as agents to mitigate neuronal damage after stroke. We developed N-(3,3-diphenylpropyl)-6-18F-fluoronicotinamide (18F-FNDP), which proved highly specific for imaging of sEH in the mouse and nonhuman primate brain with PET.
  
  
  METHODS
  18F-FNDP was synthesized from the corresponding bromo precursor. sEH inhibitory activity of18F-FNDP was measured using an sEH inhibitor screening assay kit. Biodistribution was undertaken in CD-1 mice. Binding specificity was assayed in CD-1 and sEH knock-out mice and Papio anubis (baboon) through pretreatment with an sEH inhibitor to block sEH binding. Dynamic PET imaging with arterial blood sampling was performed in 3 baboons, with regional tracer binding quantified using distribution volume. The metabolism of18F-FNDP in baboons was assessed using high-performance liquid chromatography.
  
  
  RESULTS
  18F-FNDP (inhibition binding affinity constant, 1.73 nM) was prepared in 1 step in a radiochemical yield of 14% ± 7%, specific radioactivity in the range of 888-3,774 GBq/μmol, and a radiochemical purity greater than 99% using an automatic radiosynthesis module. The time of preparation was about 75 min. In CD-1 mice, regional uptake followed the pattern of striatum > cortex > hippocampus > cerebellum, consistent with the known brain distribution of sEH, with 5.2% injected dose per gram of tissue at peak uptake. Blockade of 80%-90% was demonstrated in all brain regions. Minimal radiotracer uptake was present in sEH knock-out mice. PET baboon brain distribution paralleled that seen in mouse, with a marked blockade (95%) noted in all regions indicating sEH-mediated uptake of18F-FNDP. Two hydrophilic metabolites were identified, with 20% parent compound present at 90 min after injection in baboon plasma.
  
  
  CONCLUSIONS
  18F-FNDP can be synthesized in suitable radiochemical yield and high specific radioactivity and purity. In vivo imaging experiments demonstrated that18F-FNDP targeted sEH in murine and nonhuman primate brain specifically.18F-FNDP is a promising PET radiotracer likely to be useful for understanding the role of sEH in a variety of conditions affecting the central nervous system.
authors.affiliation: Russell H. Morgan Department of Radiology and Radiological Science, Johns Hopkins Medical Institutions, Baltimore, Maryland mpomper1@jh.edu ahorti1@jhmi.edu.
authors.forename: Andrew G
authors.initials: AG
authors.lastname: Horti
authors.affiliation: Russell H. Morgan Department of Radiology and Radiological Science, Johns Hopkins Medical Institutions, Baltimore, Maryland.
authors.forename: Yuchuan
authors.initials: Y
authors.lastname: Wang
authors.affiliation: Russell H. Morgan Department of Radiology and Radiological Science, Johns Hopkins Medical Institutions, Baltimore, Maryland.
authors.forename: Il
authors.initials: I
authors.lastname: Minn
authors.affiliation: Department of Neurology, Anesthesiology & Critical Care Medicine, Johns Hopkins Medical Institutions, Baltimore, Maryland; and.
authors.forename: Xi
authors.initials: X
authors.lastname: Lan
authors.affiliation: Department of Neurology, Anesthesiology & Critical Care Medicine, Johns Hopkins Medical Institutions, Baltimore, Maryland; and.
authors.forename: Jian
authors.initials: J
authors.lastname: Wang
authors.affiliation: Department of Neurology, Anesthesiology & Critical Care Medicine, Johns Hopkins Medical Institutions, Baltimore, Maryland; and.
authors.forename: Raymond C
authors.initials: RC
authors.lastname: Koehler
authors.affiliation: Department of Anesthesiology & Perioperative Medicine, Knight Cardiovascular Institute, Oregon Health & Science University, Portland, Oregon.
authors.forename: Nabil J
authors.initials: NJ
authors.lastname: Alkayed
authors.affiliation: Russell H. Morgan Department of Radiology and Radiological Science, Johns Hopkins Medical Institutions, Baltimore, Maryland.
authors.forename: Robert F
authors.initials: RF
authors.lastname: Dannals
authors.affiliation: Russell H. Morgan Department of Radiology and Radiological Science, Johns Hopkins Medical Institutions, Baltimore, Maryland mpomper1@jh.edu ahorti1@jhmi.edu.
authors.forename: Martin G
authors.initials: MG
authors.lastname: Pomper
mesh_terms: D000818:Animals; D001921:Brain / Q000201:enzymology*; D004851:Epoxide Hydrolases / Q000032:analysis* / Q000037:antagonists & inhibitors; D005462:Fluorine Radioisotopes; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D010215:Papio; D049268:Positron-Emission Tomography / Q000379:methods*; D019275:Radiopharmaceuticals; D014018:Tissue Distribution
chemical_list: D005462:Fluorine Radioisotopes; D019275:Radiopharmaceuticals; D004851:Epoxide Hydrolases
keywords: baboon; epoxyeicosatrienoic acid; molecular neuroimaging; positron emission tomography; soluble epoxide hydrolase; stroke; vascular cognitive impairment