In vivo assessment of aminopeptidase N (APN/CD13) specificity of different 68Ga-labelled NGR derivatives using PET/MRI imaging

Int J Pharm. 2020 Nov 15:589:119881. doi: 10.1016/j.ijpharm.2020.119881. Epub 2020 Sep 16.


Aminopeptidase N (APN/CD13) plays an important role in neoangiogenic process in malignancies. Our previous studies have already shown that 68Ga-labelled NOTA conjugated asparagine-glycine-arginine peptide (c[KNGRE]-NH2) specifically bind to APN/CD13 expressing tumors. The aim of this study was to evaluate and compare the APN/CD13 specificity of newly synthesized 68Ga-labelled NGR derivatives in vivo by PET/MRI imaging using hepatocellular carcinoma (He/De) and mesoblastic nephroma (Ne/De) tumor models. PET/MRI and ex vivo biodistribution studies were performed 11 ± 1 days after subcutaneous injection of tumor cells and 90 min after intravenous injection of 68Ga-NOTA-c(NGR), 68Ga-NODAGA-c(NGR), 68Ga-NODAGA-c(NGR) (MG1) or 68Ga-NODAGA-c(NGR) (MG2). The APN/CD13 selectivity was confirmed by blocking experiments and the APN/CD13 expression was verified by immunohistochemistry. 68Ga-labelled c(NGR) derivatives were produced with high specific activity and radiochemical purity. In control animals, low radiotracer accumulation was found in abdominal and thoracic organs. Using tumor-bearing animals we found that the 68Ga-NOTA-c(NGR), 68Ga-NODAGA-c(NGR), and 68Ga-NODAGA-c(NGR) (MG1) derivatives showed higher uptake in He/De and Ne/De tumors, than that of the accumulation of 68Ga-NODAGA-c(NGR) (MG2). APN/CD13 is a very promising target in PET imaging, however, the selection of the appropriate 68Ga-labelled NGR-based radiopharmaceutical is critical for the precise detection of tumor neo-angiogenesis and for monitoring the efficacy of anticancer therapy.

Keywords: (68)Ga; Aminopeptidase N; Angiogenesis; CD13; CID: 2796029, 1-hydroxybenzotriazole (HOBt); CID: 3036142, 1,4,7-triazacyclononane-1,4,7-triacetic acid (NOTA); CID: 33032, L-Glutamic acid; CID: 5962, L-Lysine; CID: 6228, N,N-dimethylformamide (DMF); CID: 6267, L-Asparagine; CID: 6322, L-Arginine; CID: 6422, triflouroacetic acid (TFA); CID: 750, Glyicine; NGR; PET/MRI imaging.

MeSH terms

  • Animals
  • CD13 Antigens* / metabolism
  • Cell Line, Tumor
  • Gallium Radioisotopes
  • Liver Neoplasms*
  • Magnetic Resonance Imaging
  • Male
  • Positron-Emission Tomography
  • Tissue Distribution


  • Gallium Radioisotopes
  • CD13 Antigens