Comparison of biological properties of 99mTc-labeled cyclic RGD Peptide trimer and dimer useful as SPECT radiotracers for tumor imaging

Nucl Med Biol. 2016 Nov;43(11):661-669. doi: 10.1016/j.nucmedbio.2016.02.006. Epub 2016 Feb 27.


Introduction: This study sought to evaluate a 99mTc-labeled trimeric cyclic RGD peptide (99mTc-4P-RGD3) as the new radiotracer for tumor imaging. The objective was to compare its biological properties with those of 99mTc-3P-RGD2 in the same animal model.

Methods: HYNIC-4P-RGD3 was prepared by reacting 4P-RGD3 with excess HYNIC-OSu in the presence of diisopropylethylamine. 99mTc-4P-RGD3 was prepared using a kit formulation, and evaluated for its tumor-targeting capability and biodistribution properties in the BALB/c nude mice with U87MG human glioma xenografts. Planar and SPECT imaging studies were performed in athymic nude mice with U87MG glioma xenografts. For comparison purpose, 99mTc-3P-RGD2 (a αvβ3-targeted radiotracer currently under clinical evaluation for tumor imaging in cancer patients) was also evaluated in the same animal models. Blocking experiments were used to demonstrate the αvβ3 specificity of 99mTc-4P-RGD3.

Results: 99mTc-4P-RGD3 was prepared with >95% RCP and high specific activity (~200GBq/μmol). 99mTc-4P-RGD3 and 99mTc-3P-RGD2 shared almost identical tumor uptake and similar biodistribution properties. 99mTc-4P-RGD3 had higher uptake than 99mTc-3P-RGD2 in the intestines and kidneys; but it showed better metabolic stability. The U87MG tumors were clearly visualized by SPECT with excellent contrast with 99mTc-4P-RGD3 and 99mTc-3P-RGD2.

Conclusion: Increasing peptide multiplicity from 3P-RGD2 to 4P-RGD3 offers no advantages with respect to the tumor-targeting capability. 99mTc-4P-RGD3 is as good a SPECT radiotracer as 99mTc-3P-RGD2 for imaging αvβ3-positive tumors.

Keywords: (99m)Tc-trimeric cyclic RGD peptide; SPECT; integrin α(v)β(3); tumor imaging.

Publication types

  • Comparative Study

MeSH terms

  • Animals
  • Cell Line, Tumor
  • Dimerization*
  • Drug Stability
  • Gene Expression Regulation, Neoplastic
  • Humans
  • Hydrazines / chemistry
  • Integrin alphaVbeta3 / metabolism
  • Isotope Labeling
  • Mice
  • Nicotinic Acids / chemistry
  • Peptides, Cyclic / chemistry*
  • Peptides, Cyclic / pharmacokinetics
  • Polymerization
  • Radioactive Tracers
  • Radiochemistry
  • Single Photon Emission Computed Tomography Computed Tomography / methods*
  • Technetium / chemistry*
  • Tissue Distribution


  • 6-hydrazinopyridine-3-carboxylic acid
  • Hydrazines
  • Integrin alphaVbeta3
  • Nicotinic Acids
  • Peptides, Cyclic
  • Radioactive Tracers
  • cyclic arginine-glycine-aspartic acid peptide
  • Technetium