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Evaluating Ga-68 Peptide Conjugates for Targeting VPAC Receptors: Stability and Pharmacokinetics

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Abstract

Purpose

In recent years, considerable progress has been made in the use of gallium-68 labeled receptor-specific peptides for imaging oncologic diseases. The objective was to examine the stability and pharmacokinetics of [68Ga]NODAGA and DOTA-peptide conjugate targeting VPAC [combined for vasoactive intestinal peptide (VIP) and pituitary adenylate cyclase-activating peptide (PACAP)] receptors on tumor cells.

Procedures

A VPAC receptor-specific peptide was chosen as a model peptide and conjugated to NODAGA and DOTA via solid-phase synthesis. The conjugates were characterized by HPLC and MALDI-TOF. Following Ga-68 chelation, the radiochemical purity of Ga-68 labeled peptide conjugate was determined by radio-HPLC. The stability was tested against transmetallation using 100 nM Fe3+/Zn2+/Ca2+ ionic solution and against transchelation using 200 μM DTPA solution. The ex vivo and in vivo stability of the Ga-68 labeled peptide conjugate was tested in mouse plasma and urine. Receptor specificity was determined ex vivo by cell binding assays using human breast cancer BT474 cells. Positron emission tomography (PET)/X-ray computed tomography (CT) imaging, tissue distribution, and blocking studies were performed in mice bearing BT474 xenografts.

Results

The chemical and radiochemical purity was greater than 95 % and both conjugates were stable against transchelation and transmetallation. Ex vivo stability at 60 min showed that the NODAGA-peptide-bound Ga-68 reduced to 42.1 ± 3.7 % (in plasma) and 37.4 ± 2.9 % (in urine), whereas the DOTA-peptide-bound Ga-68 was reduced to 1.2 ± 0.3 % (in plasma) and 4.2 ± 0.4 % (in urine) at 60 min. Similarly, the in vivo stability for [68Ga]NODAGA-peptide was decreased to 2.1 ± 0.2 % (in plasma) and 2.2 ± 0.4 % (in urine). For [68Ga]DOTA-peptide, it was decreased to 1.4 ± 0.3 % (in plasma) and 1.2 ± 0.4 % (in urine) at 60 min. The specific BT474 cell binding was 53.9 ± 0.8 % for [68Ga]NODAGA-peptide, 25.8 ± 1.4 % for [68Ga]-DOTA-peptide, and 18.8 ± 2.5 % for [68Ga]GaCl3 at 60 min. Inveon microPET/CT imaging at 1 h post-injection showed significantly (p < 0.05) higher tumor to muscle (T/M) ratio for [68Ga]NODAGA-peptide (3.4 ± 0.3) as compared to [68Ga]DOTA-peptide (1.8 ± 0.6). For [68Ga]GaCl3 and blocked mice, their ratios were 1.5 ± 0.6 and 1.5 ± 0.3 respectively. The tissue distributions data were similar to the PET imaging data.

Conclusion

NODAGA is superior to DOTA in terms of radiolabeling kinetics. The method of radiolabeling was reproducible and yielded higher specific activity. Although both agents have relatively low in vivo stability, PET/CT imaging studies delineated BC tumors with [68Ga]NODAGA-peptide, but not with [68Ga]DOTA-peptide.

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Acknowledgements

We thank Kim Lee for providing technical assistance on this project.

Funding

The research, in part, was supported by NIH/NCI RO1CA157372 (MLT), NIH/NCI 1S10OD012406 (MLT), and NIH/NCI S10RR23709 (MLT).

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Authors and Affiliations

  1. Departments of Radiology, Thomas Jefferson University, Philadelphia, PA, USA

    Pardeep Kumar, Sushil K. Tripathi & Mathew L. Thakur

  2. School of Chemistry and Chemical Engineering, Henan Normal University, Xinxiang, China

    C. P. Chen

  3. Departments of Biochemistry & Molecular Biology, Thomas Jefferson University, Philadelphia, PA, USA

    Eric Wickstrom

  4. Sidney Kimmel Cancer Center, Thomas Jefferson University, Philadelphia, PA, USA

    Eric Wickstrom & Mathew L. Thakur

  5. Departments of Radiation Oncology, Thomas Jefferson University, 1020 Locust Street, JAH Suite 359, Philadelphia, PA, 19107, USA

    Mathew L. Thakur

  6. Departments of Urology, Thomas Jefferson University, Philadelphia, PA, USA

    Mathew L. Thakur

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  1. Pardeep Kumar
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Correspondence to Mathew L. Thakur.

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Conflict of Interest

Mathew L. Thakur is consultant to NuView and Zevacor, Inc. No other potential conflicts of interest relevant to this article are reported.

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Kumar, P., Tripathi, S.K., Chen, C.P. et al. Evaluating Ga-68 Peptide Conjugates for Targeting VPAC Receptors: Stability and Pharmacokinetics. Mol Imaging Biol 21, 130–139 (2019). https://doi.org/10.1007/s11307-018-1207-x

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