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Development and Preclinical Evaluation of 99mTc- and 186Re-Labeled NOTA and NODAGA Bioconjugates Demonstrating Matched Pair Targeting of GRPR-Expressing Tumors

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Abstract

Purpose

The goal of this work was to develop hydrophilic gastrin-releasing peptide receptor (GRPR)-targeting complexes of the general formula fac-[M(CO)3(L)]+ [M = natRe, 99mTc, 186Re; L: NOTA for 1, NODAGA for 2] conjugated to a powerful GRPR peptide antagonist (DPhe-Gln-Trp-Ala-Val-Gly-His-Sta-Leu-NH2) via a 6-aminohexanoic acid linker.

Procedures

Metallated-peptides were prepared employing the [M(OH2)3(CO)3]+ [M = Re, 99mTc, 186Re] precursors. Re-1/2 complexes were characterized with HR-MS. IC50 studies were performed for peptides 1/2 and their respective Re-1/2 complexes in a binding assay utilizing GRPR-expressing human PC-3 prostate cancer cells and [125I]I-Tyr4-BBN as the competing ligand. The 99mTc/186Re-complexes were identified by HPLC co-injection with their Re-analogues. All tracers were challenged in vitro at 37 °C against cysteine/histidine (phosphate-buffered saline 10 mM, pH 7.4) and rat serum. Biodistribution and micro-SPECT/CT imaging of [99mTc]Tc-1/2 and [186Re]Re-2 were performed in PC-3 tumor-bearing ICR SCID mice.

Results

High in vitro receptor affinity (IC50 2–3 nM) was demonstrated for all compounds. The 99mTc/186Re-tracers were found to be hydrophilic (log D7.4 ≤ − 1.35) and highly stable. Biodistribution in PC-3 xenografted mice revealed good tumor uptake (%ID/g at 1 h: 4.3 ± 0.7 for [99mTc]Tc-1, 8.3 ± 0.9 for [99mTc]Tc-2 and 4.2 ± 0.8 for [186Re]Re-2) with moderate retention over 24 h. Rapid renal clearance was observed for [99mTc]Tc-2 and [186Re]Re-2 (> 84 % at 4 h), indicating favorable pharmacokinetics. Micro-SPECT/CT images for the 99mTc-tracers clearly visualized PC-3 tumors in agreement with the biodistribution data and with superior imaging properties found for [99mTc]Tc-2.

Conclusions

[99mTc]Tc-2 shows promise for further development as a GRPR-imaging agent. [186Re]Re-2 demonstrated very similar in vivo behavior to [99mTc]Tc-2, and further studies are therefore justified to explore the theranostic potential of our approach for targeting of GRPR-positive cancers.

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Acknowledgments

This study was funded by the University of Missouri Research Reactor (Hennkens, internal support) and the US Department of Veterans’ Affairs (Smith, VA MERIT Application 1I01BX003392). We gratefully acknowledge Mid-America Isotopes, Inc. (Ashland, MO) for the generous donation of 99Mo/99mTc generators and the support provided by the VA Biomolecular Imaging Center at the Harry S. Truman VA Hospital and the University of Missouri-Columbia. For technical expertise and contributions, we thank Mary Embree and Daniel Ampong-O’Connor (186Re production), Dr. Fabio Gallazzi (LC-ESI-MS), Dr. Brian P. Mooney (HR-ESI-MS), Susan Rottinghaus (cell culture), Lisa Watkinson and Terry Carmack (biodistribution studies), and Ashley Berendzen (micro-SPECT/CT studies).

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Authors and Affiliations

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

    George Makris, Marina Kuchuk, Charles J. Smith & Heather M. Hennkens

  2. Research Service, Harry S. Truman Memorial Veterans’ Hospital, Columbia, MO, 65201, USA

    Rajendra P. Bandari & Charles J. Smith

  3. Department of Chemistry, University of Missouri, Columbia, MO, 65211, USA

    Silvia S. Jurisson & Heather M. Hennkens

  4. Department of Radiology, University of Missouri School of Medicine, Columbia, MO, 65212, USA

    Charles J. Smith

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  1. George Makris
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Correspondence to Heather M. Hennkens.

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Conflict of Interest

Drs. Hennkens and Makris have a patent US Application 62/932,530, “Metal Chelators and Complexes” pending to The Curators of the University of Missouri. All other authors declare that they have no conflict of interest.

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Makris, G., Bandari, R.P., Kuchuk, M. et al. Development and Preclinical Evaluation of 99mTc- and 186Re-Labeled NOTA and NODAGA Bioconjugates Demonstrating Matched Pair Targeting of GRPR-Expressing Tumors. Mol Imaging Biol 23, 52–61 (2021). https://doi.org/10.1007/s11307-020-01537-1

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  • DOI: https://doi.org/10.1007/s11307-020-01537-1

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