Evaluation of copper-64 labeled AmBaSar conjugated cyclic RGD peptide for improved microPET imaging of integrin alphavbeta3 expression

Bioconjug Chem. 2010 Aug 18;21(8):1417-24. doi: 10.1021/bc900537f.


Recently, we have developed a new cage-like bifunctional chelator 4-((8-amino-3,6,10,13,16,19-hexaazabicyclo [6.6.6] icosane-1-ylamino) methyl) benzoic acid (AmBaSar) for copper-64 labeling and synthesized the positron emission tomography (PET) tracer (64)Cu-AmBaSar-RGD. In this study, we further evaluate the biological property of this new AmBaSar chelator by using (64)Cu-AmBaSar-RGD as the model compound. In vitro and in vivo stability, lipophilicity, cell binding and uptake, microPET imaging, receptor blocking experiments, and biodistribution studies of (64)Cu-AmBaSar-RGD were investigated, and the results were directly compared with the established radiotracer (64)Cu-DOTA-RGD. The (64)Cu-AmBaSar-RGD was obtained with high radiochemical yield (> or =95%) and purity (> or =99%) under mild conditions (pH 5.0-5.5 and 23-37 degrees C) in less than 30 min. For in vitro studies, the radiochemical purity of (64)Cu-AmBaSar-RGD was more than 97% in PBS or FBS and 95% in mouse serum after 24 h of incubation. The log P value of (64)Cu-AmBaSar-RGD was -2.44 +/- 0.12. For in vivo studies, (64)Cu-AmBaSar-RGD and (64)Cu-DOTA-RGD have demonstrated comparable tumor uptake at selected time points on the basis of microPET imaging. The integrin alpha(v)beta(3) receptor specificity was confirmed by blocking experiments for both tracers. Compared with (64)Cu-DOTA-RGD, (64)Cu-AmBaSar-RGD demonstrated much lower liver accumulation in both microPET imaging and biodistribution studies. Metabolic studies also directly supported the observation that (64)Cu-AmBaSar-RGD was more stable in vivo than (64)Cu-DOTA-RGD. In summary, the in vitro and in vivo evaluations of the (64)Cu-AmBaSar-RGD have demonstrated its improved Cu-chelation stability compared with that of the established tracer (64)Cu-DOTA-RGD. The AmBaSar chelator will also have general applications for (64)Cu labeling of various bioactive molecules in high radiochemical yield and high in vivo stability.

Publication types

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

MeSH terms

  • Animals
  • Benzoates* / chemistry
  • Benzoates* / pharmacokinetics
  • Bridged Bicyclo Compounds, Heterocyclic* / chemistry
  • Bridged Bicyclo Compounds, Heterocyclic* / pharmacokinetics
  • Cell Line, Tumor
  • Chelating Agents* / chemistry
  • Chelating Agents* / pharmacokinetics
  • Copper Radioisotopes* / chemistry
  • Copper Radioisotopes* / pharmacokinetics
  • Disease Models, Animal
  • Heterocyclic Compounds, 1-Ring / chemistry
  • Heterocyclic Compounds, 1-Ring / pharmacokinetics
  • Humans
  • Integrin alphaVbeta3 / biosynthesis*
  • Integrin alphaVbeta3 / metabolism
  • Mice
  • Mice, Nude
  • Molecular Conformation
  • Neoplasms, Experimental / diagnosis
  • Peptides, Cyclic* / chemistry
  • Peptides, Cyclic* / pharmacokinetics
  • Positron-Emission Tomography / methods*
  • Tissue Distribution


  • 4-((8-amino-3,6,10,13,16,19-hexaazabicyclo(6.6.6)icosane-1-ylamino)methyl)benzoic acid
  • Benzoates
  • Bridged Bicyclo Compounds, Heterocyclic
  • Chelating Agents
  • Copper Radioisotopes
  • Heterocyclic Compounds, 1-Ring
  • Integrin alphaVbeta3
  • Peptides, Cyclic
  • cyclic arginine-glycine-aspartic acid peptide
  • 1,4,7,10-tetraazacyclododecane- 1,4,7,10-tetraacetic acid