Optimization of a Collagen-Targeted PET Probe for Molecular Imaging of Pulmonary Fibrosis

There is a large unmet need for a simple, accurate, noninvasive, quantitative, and high-resolution imaging modality to detect lung fibrosis at early stage and to monitor disease progression. Overexpression of collagen is a hallmark of organ fibrosis. Here, we describe the optimization of a collagen-targeted PET probe for staging pulmonary fibrosis. Methods: Six peptides were synthesized, conjugated to a copper chelator, and radiolabeled with ⁶⁴Cu. The collagen affinity of each probe was measured in a plate-based assay. The pharmacokinetics and metabolic stability of the probes were studied in healthy rats. The capacity of these probes to detect and stage pulmonary fibrosis in vivo was assessed in a mouse model of bleomycin-induced fibrosis using PET imaging. Results: All probes exhibited affinities in the low micromolar range (1.6 μM < Kd < 14.6 μM) and had rapid blood clearance. The probes showed 2- to 8-fold-greater uptake in the lungs of bleomycin-treated mice than sham-treated mice, whereas the distribution in other organs was similar between bleomycin-treated and sham mice. The probe ⁶⁴Cu-CBP7 showed the highest uptake in fibrotic lungs and the highest target-to-background ratios. The superiority of ⁶⁴Cu-CBP7 was traced to a much higher metabolic stability compared with the other probes. The specificity of ⁶⁴Cu-CBP7 for collagen was confirmed by comparison with a nonbinding isomer. Conclusion:⁶⁴Cu-CBP7 is a promising candidate for in vivo imaging of pulmonary fibrosis.