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[68Ga]NODAGA-RGD for imaging αvβ3 integrin expression

Abstract

Purpose

A molecular target involved in the angiogenic process is the αvβ3 integrin. It has been demonstrated in preclinical as well as in clinical studies that radiolabelled RGD peptides and positron emission tomography (PET) allow noninvasive monitoring of αvβ3 expression. Here we introduce a 68Ga-labelled NOTA-conjugated RGD peptide ([68Ga]NODAGA-RGD) and compare its imaging properties with [68Ga]DOTA-RGD using small animal PET.

Methods

Synthesis of c(RGDfK(NODAGA)) was based on solid phase peptide synthesis protocols using the Fmoc strategy. The 68Ga labelling protocol was optimized concerning temperature, peptide concentration and reaction time. For in vitro characterization, partition coefficient, protein binding properties, serum stability, αvβ3 binding affinity and cell uptake were determined. To characterize the in vivo properties, biodistribution studies and microPET imaging were carried out. For both in vitro and in vivo evaluation, αvβ3-positive human melanoma M21 and αvβ3-negative M21-L cells were used.

Results

[68Ga]NODAGA-RGD can be produced within 5 min at room temperature with high radiochemical yield and purity (> 96%). In vitro evaluation showed high αvβ3 binding affinity (IC50 = 4.7 ± 1.6 nM) and receptor-specific uptake. The radiotracer was stable in phosphate-buffered saline, pH 7.4, FeCl3 solution, and human serum. Protein-bound activity after 180 min incubation was found to be 12-fold lower than for [68Ga]DOTA-RGD. Biodistribution data 60 min post-injection confirmed receptor-specific tumour accumulation. The activity concentration of [68Ga]NODAGA-RGD was lower than [68Ga]DOTA-RGD in all organs and tissues investigated, leading to an improved tumour to blood ratio ([68Ga]NODAGA-RGD: 11, [68Ga]DOTA-RGD: 4). MicroPET imaging confirmed the improved imaging properties of [68Ga]NODAGA-RGD compared to [68Ga]DOTA-RGD.

Conclusion

The introduced [68Ga]NODAGA-RGD combines easy accessibility with high stability and good imaging properties making it an interesting alternative to the 18F-labelled RGD peptides currently used for imaging αvβ3 expression.

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Acknowledgement

Bettina Sarg and Sabine Hofer, Protein Micro Analysis Facility, Biocenter, Innsbruck Medical University are acknowledged for carrying out the LC-MS analysis. We thank Alexander Staaf, Department of Pharmaceutical Chemistry, University of Innsbruck for providing analytical data concerning synthesis of the chelator. David A. Cheresh, The Scripps Research Institute, La Jolla, CA is acknowledged for providing the human melanoma M21 and M21-L cells. Parts of the studies were financially supported by the BMBF-MoBiMed grant. This work was part of COST Action BM0607 “Targeted Radionuclide Therapy”.

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Correspondence to Roland Haubner.

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Knetsch, P.A., Petrik, M., Griessinger, C.M. et al. [68Ga]NODAGA-RGD for imaging αvβ3 integrin expression. Eur J Nucl Med Mol Imaging 38, 1303–1312 (2011). https://doi.org/10.1007/s00259-011-1778-0

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  • DOI: https://doi.org/10.1007/s00259-011-1778-0

Keywords

  • 68Ga
  • NODAGA
  • RGD peptides
  • αvβ3
  • Molecular imaging
  • Angiogenesis