In vitro and in vivo evaluation of a (18)F-labeled high affinity NOTA conjugated bombesin antagonist as a PET ligand for GRPR-targeted tumor imaging

PLoS One. 2013 Dec 3;8(12):e81932. doi: 10.1371/journal.pone.0081932. eCollection 2013.

Abstract

Expression of the gastrin-releasing peptide receptor (GRPR) in prostate cancer suggests that this receptor can be used as a potential molecular target to visualize and treat these tumors. We have previously investigated an antagonist analog of bombesin (D-Phe-Gln-Trp-Ala-Val-Gly-His-Sta-Leu-NH2, RM26) conjugated to 1,4,7-triazacyclononane-N,N',N''-triacetic acid (NOTA) via a diethylene glycol (PEG2) spacer (NOTA-P2-RM26) labeled with (68)Ga and (111)In. We found that this conjugate has favorable properties for in vivo imaging of GRPR-expression. The focus of this study was to develop a (18)F-labelled PET agent to visualize GRPR. NOTA-P2-RM26 was labeled with (18)F using aluminum-fluoride chelation. Stability, in vitro binding specificity and cellular processing tests were performed. The inhibition efficiency (IC50) of the [(nat)F]AlF-NOTA-P2-RM26 was compared to that of the (nat)Ga-loaded peptide using (125)I-Tyr(4)-BBN as the displacement radioligand. The pharmacokinetics and in vivo binding specificity of the compound were studied. NOTA-P2-RM26 was labeled with (18)F within 1 h (60-65% decay corrected radiochemical yield, 55 GBq/µmol). The radiopeptide was stable in murine serum and showed high specific binding to PC-3 cells. [(nat)F]AlF-NOTA-P2-RM26 showed a low nanomolar inhibition efficiency (IC50=4.4±0.8 nM). The internalization rate of the tracer was low. Less than 14% of the cell-bound radioactivity was internalized after 4 h. The biodistribution of [(18)F]AlF-NOTA-P2-RM26 demonstrated rapid blood clearance, low liver uptake and low kidney retention. The tumor uptake at 3 h p.i. was 5.5±0.7 %ID/g, and the tumor-to-blood, -muscle and -bone ratios were 87±42, 159±47, 38±16, respectively. The uptake in tumors, pancreas and other GRPR-expressing organs was significantly reduced when excess amount of non-labeled peptide was co-injected. The low uptake in bone suggests a high in vivo stability of the Al-F bond. High contrast PET image was obtained 3 h p.i. The initial biological results suggest that [(18)F]AlF-NOTA-P2-RM26 is a promising candidate for PET imaging of GRPR in vivo.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Bombesin / antagonists & inhibitors*
  • Cell Line, Tumor
  • Cell Transformation, Neoplastic
  • Drug Stability
  • Ethylene Glycols / chemistry
  • Fluorine Radioisotopes*
  • Heterocyclic Compounds / chemistry*
  • Heterocyclic Compounds, 1-Ring
  • Humans
  • Ligands
  • Male
  • Mice
  • Oligopeptides* / chemistry
  • Oligopeptides* / pharmacokinetics
  • Oligopeptides* / pharmacology
  • Positron-Emission Tomography / methods*
  • Prostatic Neoplasms / diagnostic imaging*
  • Prostatic Neoplasms / pathology
  • Receptors, Bombesin / metabolism*

Substances

  • Ethylene Glycols
  • Fluorine Radioisotopes
  • Heterocyclic Compounds
  • Heterocyclic Compounds, 1-Ring
  • Ligands
  • Oligopeptides
  • Receptors, Bombesin
  • 1,4,7-triazacyclononane-N,N',N''-triacetic acid
  • diethylene glycol
  • Bombesin

Grants and funding

This research was financially supported by the Swedish Cancer Society (Cancerfonden) and the Swedish Research Council (Vetenskapsradet). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.