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Characterization and Evaluation of 64Cu-Labeled A20FMDV2 Conjugates for Imaging the Integrin αvβ6

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Abstract

Purpose

The integrin αvβ6 is overexpressed in a variety of aggressive cancers and serves as a prognosis marker. This study describes the conjugation, radiolabeling, and in vitro and in vivo evaluation of four chelators to determine the best candidate for 64Cu radiolabeling of A20FMDV2, an αvβ6 targeting peptide.

Procedures

Four chelators were conjugated onto PEG28-A20FMDV2 (1): 11-carboxymethyl-1,4,8,11-tetraazabicyclo[6.6.2]hexadecane-4-methanephosphonic acid (CB-TE1A1P), 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid (DOTA), 1,4,7-triazacyclononane-1,4,7-triacetic acid (NOTA), and 4,4′-((3,6,10,13,16,19-hexazazbicyclo[6.6.6]ico-sane-1,8-diylbis(aza-nediyl))bis(methylene)dibenzoic acid (BaBaSar). All peptides were radiolabeled with 64Cu in ammonium acetate buffer at pH 6 and formulated to pH 7.2 in PBS for use. The radiotracers were evaluated using in vitro cell binding and internalization assays and serum stability assays. In vivo studies conducted include blocking, biodistribution, and small animal PET imaging. Autoradiography and histology were also conducted.

Results

All radiotracers were radiolabeled in good radiochemical purity (>95 %) under mild conditions (37–50 °C for 15 min) with high specific activity (0.58–0.60 Ci/μmol). All radiotracers demonstrated αvβ6-directed cell binding (>46 %) with similar internalization levels (>23 %). The radiotracers 64Cu-CB-TE1A1P-1 and 64Cu-BaBaSar-1 showed improved specificity for the αvβ6 positive tumor in vivo over 64Cu-DOTA-1 and 64Cu-NOTA-1 (+/− tumor uptake ratios—3.82 +/- 0.44, 3.82 ± 0.41, 2.58 ± 0.58, and 1.29 ± 0.14, respectively). Of the four radiotracers, 64Cu-NOTA-1 exhibited the highest liver uptake (10.83 ± 0.1 % ID/g at 4 h).

Conclusions

We have successfully conjugated, radiolabeled, and assessed the four chelates CB-TE1A1P, DOTA, NOTA, and BaBaSar both in vitro and in vivo. However, the data suggests no clear “best candidate” for the 64Cu-radiolabeling of A20FMDV2, but instead a trade-off between the different properties (e.g., stability, selectivity, pharmacokinetics, etc.) with no obvious effects of the individual chelators.

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Acknowledgments

The authors would like to acknowledge D.L. Kukis, L. Planutyte, J.Y. Fung, M. Jiang, and S.H. Hausner for their advice and technical support.

Grant Support

This research was supported by the Office of Science, United States Department of Energy, DE-SC0002061 and NCI 5R01 CA093375.

Conflict of Interest

No conflicts of interest exist.

Author information

Authors and Affiliations

  1. Department of Biomedical Engineering, University of California Davis, Davis, CA, USA

    Lina Y. Hu, Nadine Bauer, Leah M. Knight & Julie L. Sutcliffe

  2. Department of Radiology, University of Southern California, Los Angeles, CA, USA

    Zibo Li, Shuanglong Liu & Peter S. Conti

  3. Department of Radiology, University of Pittsburgh, Pittsburgh, PA, USA

    Carolyn J. Anderson

  4. Department of Internal Medicine, Division of Hematology/Oncology, University of California Davis, School of Medicine, Research II, 4625 2nd Ave, Sacramento, CA, 95817, USA

    Julie L. Sutcliffe

  5. Center for Molecular and Genomic Imaging, University of California Davis, Davis, CA, USA

    Julie L. Sutcliffe

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  1. Lina Y. Hu
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Correspondence to Julie L. Sutcliffe.

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Hu, L.Y., Bauer, N., Knight, L.M. et al. Characterization and Evaluation of 64Cu-Labeled A20FMDV2 Conjugates for Imaging the Integrin αvβ6 . Mol Imaging Biol 16, 567–577 (2014). https://doi.org/10.1007/s11307-013-0717-9

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  • DOI: https://doi.org/10.1007/s11307-013-0717-9

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