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A new 68Ga-labeled BBN peptide with a hydrophilic linker for GRPR-targeted tumor imaging

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Abstract

Bombesin (BBN) is a peptide exhibiting high affinity for the gastrin-releasing peptide receptor (GRPR), which is overexpressed on several types of cancers. Various GRPR antagonists and agonists have been labeled with radiometals for positron emission tomography (PET) imaging of GRPR-positive tumors. However, unfavorable hepatobiliary excretion such as high intestinal activity may prohibit their clinical utility for imaging abdominal cancer. In this study, the modified BBN peptide with a new hydrophilic linker was labeled with 68Ga for PET imaging of GRPR-expressing PC-3 prostate cancer xenograft model. GRPR antagonists, MATBBN (Gly-Gly-Gly-Arg-Asp-Asn-d-Phe-Gln-Trp-Ala-Val-Gly-His-Leu-NHCH2CH3) and ATBBN (d-Phe-Gln-Trp-Ala-Val-Gly-His-Leu-NHCH2CH3), were conjugated with 1,4,7-triazacyclononanetriacetic acid (NOTA) and labeled with 68Ga. Partition coefficient and in vitro stability were also determined. GRPR binding affinity of both tracers was investigated by competitive radioligand binding assay. The in vivo receptor targeting potential and pharmacokinetic of 68Ga-NOTA-MATBBN were also evaluated in PC-3 prostate tumor model and compared with those of 68Ga-NOTA-ATBBN. NOTA-conjugated BBN analogs were labeled with 68Ga within 20 min with a decay-corrected yield ranging from 90 to 95 % and a radiochemical purity of more than 98 %. The specific activity of 68Ga-NOTA-MATBBN and 68Ga-NOTA-ATBBN was at least 16.5 and 11.9 GBq/μmol, respectively. The radiotracers were stable in phosphate-buffered saline and human serum. 68Ga-NOTA-MATBBN was more hydrophilic than 68Ga-NOTA-ATBBN, as indicated by their log P values (−2.73 ± 0.02 vs. −1.20 ± 0.03). The IC50 values of NOTA-ATBBN and NOTA-MATBBN were similar (102.7 ± 1.18 and 124.6 ± 1.21 nM). The accumulation of 68Ga-labeled GRPR antagonists in the subcutaneous PC-3 tumors could be visualized via small animal PET. The tumors were clearly visible, and the tumor uptakes of 68Ga-NOTA-MATBBN and 68Ga-NOTA-ATBBN were determined to be 4.19 ± 0.32, 4.00 ± 0.41, 2.93 ± 0.35 and 4.70 ± 0.40, 4.10 ± 0.30, 3.14 ± 0.30 %ID/g at 30, 60, and 120 min, respectively. There was considerable accumulation and retention of 68Ga-NOTA-ATBBN in the liver and intestines. In contrast, the abdominal area does not have much retention of 68Ga-NOTA-MATBBN. Biodistribution data were in accordance with the PET results, showing that 68Ga-NOTA-MATBBN had more favorable pharmacokinetics and higher tumor to background ratios than those of 68Ga-NOTA-ATBBN. At 1 h postinjection, the tumor to liver and intestine of 68Ga-NOTA-MATBBN were 8.05 ± 0.56 and 21.72 ± 3.47 and the corresponding values of unmodified counterpart were 0.85 ± 0.23 and 3.45 ± 0.43, respectively. GRPR binding specificity was demonstrated by reduced tumor uptake of radiolabeled tracers after coinjection of an excess of unlabeled BBN peptides. 68Ga-NOTA-MATBBN exhibited GRPR-targeting properties both in vitro and in vivo. The favorable characterizations of 68Ga-NOTA-MATBBN such as convenient synthesis, specific GRPR targeting, high tumor uptake, and satisfactory pharmacokinetics warrant its further investigation for clinical cancer imaging.

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Acknowledgments

This work was partially supported by National Natural Science Foundation (81171399 and 81101077), CSC Foundation (2011832173), National Significant New Drugs Creation Program (2012ZX09505-001-001), Jiangsu Province Science and Technology Foundation (BE2012622, BK2011166 and BL2012031), Health Ministry of Jiangsu Province Fund (RC2011095 and H201028), Public Service Platform for Science and Technology Infrastructure Construction Project of Jiangsu Province (BM2012066), and University of Wisconsin-Madison Department of Medical Physics and Department of Radiology (Radiology R&D Award 1105-002).

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The authors declare that they have no conflict of interest.

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Correspondence to Min Yang or Yongjun Yan.

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Pan, D., Xu, Y.P., Yang, R.H. et al. A new 68Ga-labeled BBN peptide with a hydrophilic linker for GRPR-targeted tumor imaging. Amino Acids 46, 1481–1489 (2014). https://doi.org/10.1007/s00726-014-1718-y

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  • DOI: https://doi.org/10.1007/s00726-014-1718-y

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