(68)Ga-NOTA-c(NGR)

Elevated levels of gelatinases (matrix metalloproteinases 2/9, i.e., MMP2 and MMP9) are associated with tumor progression, invasion and metastasis, so these enzymes are potential targets for tumor imaging. The peptide c(KAHWGFTLD)NH2 (herein, C6) is a selective gelatinase inhibitor. Cy5.5-C6 has been visualized in many tumor models in vivo. However, the sensitivity and penetrance of optical imaging are poor. It is well known that positron emission tomography (PET) has a high detection sensitivity and Gallium-68 ((68)Ga) is an optimal PET radioisotope. Thus, in the present study, we developed a novel ligand, (68)Ga-NOTA-C6, to image MMP2 activity in tumors.

C6 was conjugated with the bifunctional chelator NOTA (1,4,7-triazacyclononanetriacetic acid) and labeled with (68)Ga. In vitro uptake and binding analyses were performed by using SKOV3 cell lines, coincubating with or without the MMP inhibitor doxycycline. The biodistribution and PET imaging were conducted on SKOV3 ovarian tumor models. MMP2 expression in tumors was analyzed by immunohistochemistry (IHC).

The non-decay corrected yield of (68)Ga-NOTA-C6 was 61.8%-63.3%. (68)Ga-NOTA-C6 was stable in both physiological saline and human serum. The uptake of (68)Ga-NOTA-C6 in SKOV3 cells increased with time, and could be blocked by doxycycline in a dose dependent manner. The results of biodistribution and PET imaging showed that high radioactivity concentrations of (68)Ga-NOTA-C6 occurred in tumors. The ratios of tumor to blood, muscle and ovary and oviduct at 30, 60 and 120min p.i. were 2.78±0.54, 3.86±0.65, 0.48±0.14, and 1.73±0.36, 10.31±3.12, 1.22±0.10, and 2.50±0.78, 7.03±1.85, 0.97±0.25, respectively. The tracer was excreted mainly through the renal system, as evidenced by high levels of radioactivity in the kidneys. These data support the possibility of using (68)Ga-NOTA-C6 in PET to visualize tumors that overexpress MMP2.

(68)Ga-NOTA-C6 is a potential radiopharmaceutical for the imaging of in vivo MMP2 activity in tumors.