Click Chemistry Enables [89Zr]Zr-DOTA Radioimmunoconjugation for Theranostic 89Zr-immunoPET

Bioconjug Chem. 2024 Nov 20;35(11):1744-1754. doi: 10.1021/acs.bioconjchem.4c00274. Epub 2024 Aug 16.

Abstract

There have been predictions that the use of the macrocyclic chelating agent 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid (DOTA) in zirconium-89 (89Zr) immuno-positron emission tomography (89Zr-immunoPET) could enhance the in vivo stability of 89Zr radioimmunoconjugates. However, conjugating [89Zr]Zr-DOTA to a monoclonal antibody (mAb) remains a challenge as the heat treatment required for [89Zr]Zr-DOTA chelation can lead to thermal denaturation of the mAb moieties. We developed a method for synthesizing [89Zr]Zr-DOTA-mAb based on a tetrazine (Tz)-conjugated bifunctional DOTA derivative 2,2',2″-(10-(1-(4-(1,2,4,5-tetrazin-3-yl)phenyl)-3,21,26-trioxo-6,9,12,15,18-pentaoxa-29-carboxy-2,22,25-triazanonacosane-29-yl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (DOTAGA-Tz) and the inverse electron-demand Diels-Alder (IEDDA) click chemistry reaction where trans-cyclooctene-modified mAbs are conjugated to [89Zr]Zr-DOTAGA without being exposed to heat. The stability of IEDDA-derived [89Zr]Zr-DOTAGA-trastuzumab was confirmed by in vitro, ex vivo, and in vivo testing and comparative analysis against the conventional deferoxamine (DFO) counterpart [89Zr]Zr-DFO-trastuzumab. The in vivo immunoPET imaging using [89Zr]Zr-DOTAGA-trastuzumab clearly visualized human epidermal growth factor receptor 2-positive malignancies in murine xenograft models. Greater tumor contrast was observed from [89Zr]Zr-DOTAGA-trastuzumab at a 72-h delayed scan compared with [89Zr]Zr-DFO-trastuzumab. These findings suggest that our IEDDA ligation approach can be an effective means of synthesizing [89Zr]Zr-DOTA-mAb and can enhance the theranostic potential of 89Zr-immunoPET in DOTA-mediated radioimmunotherapy.

MeSH terms

  • Animals
  • Antibodies, Monoclonal / chemistry
  • Cell Line, Tumor
  • Click Chemistry*
  • Heterocyclic Compounds, 1-Ring* / chemistry
  • Humans
  • Immunoconjugates* / chemistry
  • Mice
  • Mice, Nude
  • Positron-Emission Tomography* / methods
  • Radioimmunotherapy / methods
  • Radioisotopes* / chemistry
  • Radiopharmaceuticals / chemistry
  • Theranostic Nanomedicine / methods
  • Zirconium* / chemistry

Substances

  • Zirconium
  • Immunoconjugates
  • Zirconium-89
  • Radioisotopes
  • Heterocyclic Compounds, 1-Ring
  • 1,4,7,10-tetraazacyclododecane- 1,4,7,10-tetraacetic acid
  • Antibodies, Monoclonal
  • Radiopharmaceuticals