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Synthesis and Application of a Long-Circulating Radiolabeled Peptide for Targeting of Osteosarcoma

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The small peptide TMTP1 (NVVRQ) has been proved to target a series of highly metastatic tumor cells. The aim of this study was to develop a new agent based on TMTP1 conjugated with Evans blue (EB), to increase tumor uptake and modify the pharmacokinetic characteristics of the resulting radiolabeled agent.


DOTA-EB-TMTP1 was prepared through conventional solid-phase peptide synthesis chemistry. Then, it was successfully labeled with Cu-64 to obtain [64Cu]DOTA-EB-TMTP1. The tumor targeting properties were evaluated in vivo using 143B xenografts.


DOTA-EB-TMTP1 was successfully labeled with Cu-64 in a yield of 87.3 ± 5.2 %. In a small animal positron emission tomography/X-ray computed tomography (PET/CT) study in osteosarcoma 143B xenograft mice, [64Cu]DOTA-EB-TMTP1 was found to rapidly accumulate in the tumor tissue. The tumor uptake increased over time and reached a plateau of 6.50 ± 0.88 % ID/g 8 h after tail vein injection. The radioactivity remained in the tumor tissue 48 h postinjection with a negligible decrease.


Overall, the introduction of the EB motif to TMTP1 significantly changed its pharmacokinetics in vivo, and this strategy fulfills the purpose of prolonging the blood circulation and enhancing the tumor uptake. [64Cu]DOTA-EB-TMTP1 is a promising agent for osteosarcoma targeting. Moreover, our study highlights that DOTA-EB-TMTP1 is a good candidate for labeling with different radionuclides for potential theranostic applications.

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Yesen Li was funded by the China Scholarship Council (CSC). We thank Drs. Yanjiang Han, Bulin Du, and Frezghi G. Habte for their assistance in the imaging study.


This study was funded by the National Natural Science Foundation of China (81701762) and Natural Science Foundation of Fujian Province of China (No. 2016J01642). Funding support was provided in part by the Stanford University, Department of Radiology.

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Correspondence to Jinxiong Huang or Zhen Cheng.

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All animal procedures were performed in accordance with the Guide for the Care and Use of Laboratory Animals (Xiamen University Institutional Animal Care and Use Committee).

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Li, Y., Li, D., Wu, H. et al. Synthesis and Application of a Long-Circulating Radiolabeled Peptide for Targeting of Osteosarcoma. Mol Imaging Biol (2020).

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Key words

  • TMTP1
  • Evans blue
  • Cu-64
  • Osteosarcoma
  • PET