Improved Radiolytic Stability of a 68Ga-labelled Collagelin Analogue for the Imaging of Fibrosis

Irina Velikyan; Ulrika Rosenström; Maria Rosestedt; Olof Eriksson; Gunnar Antoni

doi:10.3390/ph14100990

Improved Radiolytic Stability of a ⁶⁸Ga-labelled Collagelin Analogue for the Imaging of Fibrosis

Pharmaceuticals (Basel). 2021 Sep 28;14(10):990. doi: 10.3390/ph14100990.

Authors

Irina Velikyan^{1

2}, Ulrika Rosenström³, Maria Rosestedt¹, Olof Eriksson¹, Gunnar Antoni^{1

2}

Affiliations

¹ Science for Life Laboratory, Department of Medicinal Chemistry, Uppsala University, SE-75183 Uppsala, Sweden.
² PET-Centre, Centre for Medical Imaging, Uppsala University Hospital, SE-75185 Uppsala, Sweden.
³ Department of Medicinal Chemistry, Uppsala University, SE-75123 Uppsala, Sweden.

Abstract

There is an unmet medical need for non-invasive, sensitive, and quantitative methods for the assessment of fibrosis. Herein, an improved collagelin analogue labelled with gallium-68 for use with positron emission tomography (PET) is presented. A cyclic peptide, c[CPGRVNleHGLHLGDDEGPC], was synthesized by solid-phase peptide synthesis, conjugated to 2-(4,7-bis(2-(tert-butoxy)-2-oxoethyl)-1,4,7-triazonan-1-yl)acetic acid, and labelled with gallium-68. High performance liquid chromatography (HPLC) was used for the quality and stability assessment of the collagelin analogue. Non-specific organ distribution, blood clearance, and excretion rates were investigated in healthy mice and rats using ex vivo organ distribution analysis and dynamic in vivo PET/CT. Mice with carbon tetrachloride (CCl₄) induced liver fibrosis were used for the investigation of specific binding via in vitro frozen section autoradiography, ex vivo organ distribution, and in vivo PET/CT. A non-decay corrected radiochemical yield (48 ± 6%) of [⁶⁸Ga]Ga-NOTA-PEG₂-c[CPGRVNleHGLHLGDDEGPC] ([⁶⁸Ga]Ga-NO2A-[Nle¹³]-Col) with a radiochemical purity of 98 ± 2% was achieved without radical scavengers. The ⁶⁸Ga-labelling was regioselective and stable at ambient temperature for at least 3 h. The autoradiography of the cryosections of fibrotic mouse liver tissue demonstrated a distinct heterogeneous radioactivity uptake that correlated with the fibrosis scores estimated after Sirius Red staining. The blood clearance and tissue washout from the [⁶⁸Ga]Ga-NO2A-[Nle¹³]-Col was fast in both normal and diseased mice. Dosimetry investigation in rats indicated the possibility for 4-5 PET/CT examinations per year. Radiolytic stability of the collagelin analogue was achieved by the substitution of methionine with norleucine amino acid residue without a deterioration of its binding capability. [⁶⁸Ga]Ga-NO2A-[Nle¹³]-Col demonstrated a safe dosimetry profile suitable for repeated scanning.

Keywords: collagelin; collagen; fibrosis; gallium-68; molecular imaging; positron emission tomography; radiochemistry.