<i>p</i>-NO₂–Bn–H₄neunpa and H₄neunpa–Trastuzumab: Bifunctional Chelator for Radiometalpharmaceuticals and ¹¹¹In Immuno-Single Photon Emission Computed Tomography Imaging

Abstract

Potentially nonadentate (N₅O₄) bifunctional chelator p-SCN-Bn-H₄neunpa and its immunoconjugate H₄neunpa-trastuzumab for ¹¹¹In radiolabeling are synthesized. The ability of p-SCN-Bn-H₄neunpa and H₄neunpa-trastuzumab to quantitatively radiolabel ¹¹¹InCl₃ at an ambient temperature within 15 or 30 min, respectively, is presented. Thermodynamic stability determination with In³⁺, Bi³⁺, and La³⁺ resulted in high conditional stability constant (pM) values. In vitro human serum stability assays have demonstrated both ¹¹¹In complexes to have high stability over 5 days. Mouse biodistribution of [¹¹¹In][In(p-NO₂-Bn-neunpa)]^-, compared to that of [¹¹¹In][In(p-NH₂-Bn-CHX-A″-diethylenetriamine pentaacetic acid (DTPA))]^2-, at 1, 4, and 24 h shows fast clearance of both complexes from the mice within 24 h. In a second mouse biodistribution study, the immunoconjugates ¹¹¹In-neunpa-trastuzumab and ¹¹¹In-CHX-A″-DTPA-trastuzumab demonstrate a similar distribution profile but with slightly lower tumor uptake of ¹¹¹In-neunpa-trastuzumab compared to that of ¹¹¹In-CHX-A″-DTPA-trastuzumab. These results were also confirmed by immuno-single photon emission computed tomography (immuno-SPECT) imaging in vivo. These initial investigations reveal the acyclic bifunctional chelator p-SCN-Bn-H₄neunpa to be a promising chelator for ¹¹¹In (and other radiometals) with high in vitro stability and also show H₄neunpa-trastuzumab to be an excellent ¹¹¹In chelator with promising biodistribution in mice.