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Preliminary evaluation of 177Lu-labeled knottin peptides for integrin receptor-targeted radionuclide therapy

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European Journal of Nuclear Medicine and Molecular Imaging Aims and scope Submit manuscript

Abstract

Purpose

Cystine knot peptides (knottins) 2.5D and 2.5F were recently engineered to bind integrin receptors with high affinity and specificity. These receptors are overexpressed on the surface of a variety of malignant human tumor cells and tumor neovasculature. In this study, 2.5D and 2.5F were labeled with a therapeutic radionuclide, 177Lu, and the resulting radiopeptides were then evaluated as potential radiotherapeutic agents in a murine model of human glioma xenografts.

Methods

Knottins 2.5D and 2.5F were synthesized using solid phase peptide synthesis, folded in vitro, and site-specifically coupled with 1,4,7,10-tetraazacyclododecane-N,N′,N′′,N′′′-tetraacetic acid (DOTA) at their N terminus for 177Lu radiolabeling. The stability of the radiopeptides 177Lu-DOTA-2.5D and 177Lu-DOTA-2.5F was tested in both phosphate-buffered saline (PBS) and mouse serum. Cell uptake assays of the radiolabeled peptides were performed in U87MG integrin-expressing human glioma cells. The biodistribution studies of both 177Lu-DOTA-2.5D and 177Lu-DOTA-2.5F were examined in U87MG tumor-bearing athymic nu/nu mice. Radiation absorbed doses for the major tissues of a human adult male were calculated based on the mouse biodistribution results.

Results

DOTA-2.5D and DOTA-2.5F were labeled with 177Lu at over 55% efficiency. High radiochemical purity for both radiocomplexes (> 95%) could be achieved after high performance liquid chromatography (HPLC) purification. Both radiopeptides were stable in PBS and mouse serum. Compared to 177Lu-DOTA-2.5D (0.39 and 0.26 %ID/g at 2 and 24 h, respectively), 177Lu-DOTA-2.5F showed much higher tumor uptake (2.16 and 0.78 %ID/g at 2 and 24 h, respectively). It also displayed higher tumor to blood ratios than that of 177Lu-DOTA-2.5D (31.8 vs 18.7 at 24 h and 52.6 vs 20.6 at 72 h). Calculation of radiodosimetry for 177Lu-DOTA-2.5D and 177Lu-DOTA-2.5F suggested that tumor and kidney were tissues with the highest radiation absorbed doses. Moreover, 177Lu-DOTA-2.5F had a higher tumor to kidney radiation absorbed dose ratio than that of 177Lu-DOTA-2.5D.

Conclusion

Cystine knot peptides can be successfully radiolabeled with 177Lu for potential therapeutic applications. Knottin 2.5F labeled with 177Lu exhibits favorable distribution in murine U87MG xenograft model; thus, it is a promising agent for radionuclide therapy of integrin-positive tumors.

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Acknowledgements

This work was supported, in part, by the National Cancer Institute (NCI) 5R01 CA119053 (ZC), and the In Vivo Cellular Molecular Imaging Center (ICMIC) grant P50 CA114747 (SSG), the Edward Mallinckrodt Jr. Foundation (JRC), as well as a Ph.D. student fellowship from the China Scholarship Council (LJ).

Conflicts of interest

None.

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Authors and Affiliations

  1. Department of Nuclear Medicine, Shanghai Ruijin Hospital, Shanghai Jiaotong University, 2nd Ruijin Rd, Shanghai, 200025, People’s Republic of China

    Lei Jiang & Peiyong Li

  2. Molecular Imaging Program at Stanford (MIPS), Department of Radiology, Stanford Cancer Center, Bio-X Program, Stanford University, 1201 Welch Road, Lucas Expansion, P020A, Stanford, CA, 94305, USA

    Lei Jiang, Zheng Miao, Hongguang Liu & Zhen Cheng

  3. Department of Bioengineering, Stanford Cancer Center, Bio-X Program, Stanford University, Stanford, CA, 94305, USA

    Richard H. Kimura & Jennifer R. Cochran

  4. Research Reactor Center, University of Missouri, Columbia, MO, 65211, USA

    Cathy S. Culter

  5. Departments of Radiology and Otolaryngology – Head and Neck Surgery, University of Texas Health Science Center at San Antonio, San Antonio, TX, 78229, USA

    Ande Bao

Authors
  1. Lei Jiang
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  2. Zheng Miao
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  3. Richard H. Kimura
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  4. Hongguang Liu
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  5. Jennifer R. Cochran
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  7. Ande Bao
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  8. Peiyong Li
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  9. Zhen Cheng
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Corresponding authors

Correspondence to Peiyong Li or Zhen Cheng.

Additional information

Lei Jiang and Zheng Miao contributed equally to the work.

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Jiang, L., Miao, Z., Kimura, R.H. et al. Preliminary evaluation of 177Lu-labeled knottin peptides for integrin receptor-targeted radionuclide therapy. Eur J Nucl Med Mol Imaging 38, 613–622 (2011). https://doi.org/10.1007/s00259-010-1684-x

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  • DOI: https://doi.org/10.1007/s00259-010-1684-x

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