Synthesis and Evaluation of Nicotine alpha4beta2 Receptor Radioligand, 5-(3'-18F-fluoropropyl)-3-(2-(S)-pyrrolidinylmethoxy)pyridine, in Rodents and PET in Nonhuman Primate

J Nucl Med. 2005 Jan;46(1):130-40.


Nicotine alpha(4)beta(2) receptor subtypes are implicated in the study of Alzheimer's disease, schizophrenia, substance abuse, lung cancer, and other disorders. We report the development and evaluation of a putative antagonist, 5-(3'-fluoropropyl)-3-(2-(S)-pyrrolidinylmethoxy)pyridine (nifrolidine) as a PET agent for nicotine alpha(4)beta(2) receptors.

Methods: In vitro binding affinity of nifrolidine was measured in rat brain slices labeled with (125)I-iodoepibatidine or (125)I-bungaratoxin. Selectivity of binding was measured in the presence of cytisine. (18)F radiolabeling was performed by reacting the tosylate precursor with (18)F-fluoride followed by deprotection. In vitro autoradiographic studies in rat brain slices with 5-(3'-(18)F-fluoropropyl)-3-(2-(S)-pyrrolidinylmethoxy)pyridine ((18)F-nifrolidine) were read on a phosphor imager. Rats were injected with (18)F-nifrolidine (3.7 MBq each), and brain regions were counted at various times (2-120 min). Blocking studies were performed by subcutaneous injection of nicotine (10 mg/kg). A PET study of (18)F-nifrolidine (approximately 148 MBq) was performed on an anesthetized rhesus monkey using a high-resolution scanner.

Results: In vitro binding affinity of nifrolidine exhibited an inhibition constant of 2.89 nmol/L for the alpha(4)beta(2) sites. Radiosynthesis and high-performance liquid chromatography purifications yielded the product in approximately 20%-40% decay-corrected radiochemical yield to provide (18)F-nifrolidine specific activities of approximately 111-185 GBq/mumol. In vitro autoradiography in rat brain slices revealed selective binding of (18)F-nifrolidine to the anteroventral thalamic nucleus, ventral posteriomedial thalamus, dorsolateral geniculate, and, to a lesser extent, cortex and striata, which are known to contain alpha(4)beta(2) sites. This specific binding was completely abolished by 300 mumol/L nicotine. Ex vivo rat brain distribution studies indicated selective binding in the thalamus with a maximal thalamus-to-cerebellum ratio of approximately 3. The PET study revealed selective maximal uptake (0.01% injected dose/mL) in regions of the thalamus (anteroventral and anteromedial thalamus, ventrolateral thalamus) and extrathalamic regions such as cingulate gyrus, lateral geniculate, temporal cortex, and frontal cortex.

Conclusion: Binding of (18)F-nifrolidine to alpha(4)beta(2) receptor-rich regions in rats and monkeys indicates promise as a PET agent. Additionally, the thalamus-to-cerebellum ratio approached a plateau of 1.7 in 120 min, indicating relatively faster kinetics compared with previously reported imaging agents.

Publication types

  • Evaluation Study
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Brain / diagnostic imaging*
  • Brain / metabolism*
  • Bridged Bicyclo Compounds, Heterocyclic / chemistry
  • Bridged Bicyclo Compounds, Heterocyclic / pharmacokinetics*
  • Isotope Labeling / methods
  • Macaca mulatta
  • Male
  • Metabolic Clearance Rate
  • Positron-Emission Tomography
  • Protein Binding
  • Pyridines / chemistry
  • Pyridines / pharmacokinetics*
  • Pyrrolidines / chemistry
  • Pyrrolidines / pharmacokinetics*
  • Radiopharmaceuticals / chemical synthesis
  • Radiopharmaceuticals / pharmacokinetics
  • Rats
  • Rats, Sprague-Dawley
  • Receptors, Nicotinic / metabolism*
  • Tissue Distribution


  • 2-(6-iodo-3-pyridinyl)-7-azabicyclo(2.2.1)heptane
  • 5-(3'-fluoropropyl)-3-(2-pyrrolidinylmethoxy)pyridine
  • Bridged Bicyclo Compounds, Heterocyclic
  • Pyridines
  • Pyrrolidines
  • Radiopharmaceuticals
  • Receptors, Nicotinic
  • nicotinic receptor alpha4beta2