title: Evaluation of an integrin αvβ6-specific peptide labeled with [18F]fluorine by copper-free, strain-promoted click chemistry.
abstract: BACKGROUND
  Click chemistry, particularly the Huisgen 1,3-dipolar cycloaddition of an alkyne with an azide, has quickly become popular for site-specific radiolabeling. Recently, strain-promoted click chemistries have been developed, eliminating the need for potentially toxic copper catalysts. This study presents radiolabeling of an α(v)β(6) integrin targeting peptide (A20FMDV2) via strain-promoted click using a fluorine-18 prosthetic group, and in vitro and in vivo evaluation.
  
  
  METHODS
  The radiotracer was prepared from and N(3)-PEG(7)-A20FMDV2 (ethanol; 10 min; 35-45 °C). HPLC-purified and formulated radiotracer 1 was evaluated in vitro by cell binding (DX3puroβ6, α(v)β(6)-positive; and DX3puro, α(v)β(6)-negative control) and serum stability, and in vivo using PET/CT imaging and biodistribution studies in mice.
  
  
  RESULTS
  The radiotracer 1 was readily prepared and purified (from 2: 40±4 min including HPLC, 11.9±3.2% decay corrected isolated radiochemical yield, >99% radiochemical purity, n=4) and displayed good stability (1 h: >99%, saline; 94.6%, serum). Strong α(v)β(6)-targeted binding was observed in vitro (DX3puroβ6 cells, 15 min: 43.2% binding, >6:1 for DX3puroβ6:DX3puro). In the mouse model DX3puroβ6-tumor binding was low (1 h: 0.47±0.28% ID/g, 4h: 0.14±0.09% ID/g) and clearing from the bloodstream was via the renal and hepatobiliary routes (urine: 167±84% ID/g at 1 h, 10.3±4.8% ID/g at 4 h; gall bladder: 95±33% ID/g at 1 h, 63±11% ID/g at 4 h).
  
  
  CONCLUSIONS
  Copper-free, strain-promoted click chemistry is an attractive, straightforward approach to radiolabeling. Although the [(18)F]FBA-C(6)-ADBIO-based prosthetic group did not interfere with α(v)β(6)-targeted binding in vitro, it did influence the pharmacokinetics, possibly due to its size and lipophilic nature.
authors.affiliation: Department of Biomedical Engineering, University of California Davis, Davis, CA 95616, USA.
authors.forename: Sven H
authors.initials: SH
authors.lastname: Hausner
authors.forename: Richard D
authors.initials: RD
authors.lastname: Carpenter
authors.forename: Nadine
authors.initials: N
authors.lastname: Bauer
authors.forename: Julie L
authors.initials: JL
authors.lastname: Sutcliffe
mesh_terms: D000818:Animals; D000951:Antigens, Neoplasm / Q000378:metabolism*; D002460:Cell Line; D057930:Click Chemistry / Q000379:methods*; D005260:Female; D005462:Fluorine Radioisotopes / Q000737:chemistry*; D016023:Integrins / Q000378:metabolism*; D007553:Isotope Labeling; D008297:Male; D051379:Mice; D010455:Peptides / Q000737:chemistry* / Q000378:metabolism* / Q000493:pharmacokinetics; D011092:Polyethylene Glycols / Q000737:chemistry; D011854:Radiochemistry / Q000379:methods*; D051381:Rats
chemical_list: D000951:Antigens, Neoplasm; D005462:Fluorine Radioisotopes; D016023:Integrins; D010455:Peptides; C073878:integrin alphavbeta6; D011092:Polyethylene Glycols