Development of a 99mTc-labeled tetrazine for pretargeted SPECT imaging using an alendronic acid-based bone targeting model

PLoS One. 2024 Apr 16;19(4):e0300466. doi: 10.1371/journal.pone.0300466. eCollection 2024.

Abstract

Pretargeting, which is the separation of target accumulation and the administration of a secondary imaging agent into two sequential steps, offers the potential to improve image contrast and reduce radiation burden for nuclear imaging. In recent years, the tetrazine ligation has emerged as a promising approach to facilitate covalent pretargeted imaging due to its unprecedented kinetics and bioorthogonality. Pretargeted bone imaging with TCO-modified alendronic acid (Aln-TCO) is an attractive model that allows the evaluation of tetrazines in healthy animals without the need for complex disease models or targeting regimens. Recent structure-activity relationship studies of tetrazines evaluated important parameters for the design of potent tetrazine-radiotracers for pretargeted imaging. However, limited information is available for 99mTc-labeled tetrazines. In this study, four tetrazines intended for labeling with fac-[99mTc(OH2)3 (CO)3]+ were synthesized and evaluated using an Aln-TCO mouse model. 3,6-bis(2-pyridyl)-1,2,4,5-Tz without additional linker showed higher pretargeted bone uptake and less background activity compared to the same scaffold with a PEG8 linker or 3-phenyl-1,2,4,5-Tz-based compounds. Additionally, improved bone/blood ratios were observed in pretargeted animals compared to animals receiving directly labeled Aln-TCO. The results of this study implicate 3,6-bis(2-pyridyl)-1,2,4,5-Tz as a promising scaffold for potential 99mTc-labeled tetrazines.

MeSH terms

  • Animals
  • Cell Line, Tumor
  • Heterocyclic Compounds*
  • Mice
  • Positron-Emission Tomography / methods
  • Radiopharmaceuticals
  • Tomography, Emission-Computed, Single-Photon / methods
  • Tomography, X-Ray Computed*

Substances

  • Heterocyclic Compounds
  • Radiopharmaceuticals

Grants and funding

This research was made possible by grants from the Lundbeck Foundation of Denmark (Lundbeck Foundation Professorship to UOH, No. 2014-4176), the Natural Sciences and Engineering Research Council of Canada (NSERC, Discovery Grant to UOH, No. 2018-04958) and the Canada Foundation for Innovation (Project No. 25413) for its support of the imaging facility (http://invivoimaging.ca/). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.