[²¹³Bi]Bi³⁺/[¹¹¹In]In³⁺-neunpa-cycMSH: Theranostic Radiopharmaceutical Targeting Melanoma─Structural, Radiochemical, and Biological Evaluation

Abstract

With the emergence of [²²⁵Ac]Ac³⁺ as a therapeutic radionuclide for targeted α therapy (TAT), access to clinical quantities of the potent, short-lived α-emitter [²¹³Bi]Bi³⁺ (<i>t</i>_1/2 = 45.6 min) will increase over the next decade. With this in mind, the nonadentate chelator, H₄neunpa-NH₂, has been investigated as a ligand for chelation of [²¹³Bi]Bi³⁺ in combination with [¹¹¹In]In³⁺ as a suitable radionuclidic pair for TAT and single photon emission computed tomography (SPECT) diagnostics. Nuclear magnetic resonance (NMR) spectroscopy was utilized to assess the coordination characteristics of H₄neunpa-NH₂ on complexation of [^natBi]Bi³⁺, while the solid-state structure of [^natBi][Bi(neunpa-NH₃)] was characterized via X-ray diffraction (XRD) studies, and density functional theory (DFT) calculations were performed to elucidate the conformational geometries of the metal complex in solution. H₄neunpa-NH₂ exhibited fast complexation kinetics with [²¹³Bi]Bi³⁺ at RT achieving quantitative radiolabeling within 5 min at 10^-8 M ligand concentration, which was accompanied by the formation of a kinetically inert complex. Two bioconjugates incorporating the melanocortin 1 receptor (MC1R) targeting peptide Nle-CycMSH_hex were synthesized featuring two different covalent linkers for <i>in vivo</i> evaluation with [²¹³Bi]Bi³⁺ and [¹¹¹In]In³⁺. High molar activities of 7.47 and 21.0 GBq/μmol were achieved for each of the bioconjugates with [²¹³Bi]Bi³⁺. SPECT/CT scans of the [¹¹¹In]In³⁺-labeled tracer showed accumulation in the tumor over time, which was accompanied by high liver uptake and clearance via the hepatic pathway due to the high lipophilicity of the covalent linker. <i>In vivo</i> biodistribution studies in C57Bl/6J mice bearing B16-F10 tumor xenografts showed good tumor uptake (5.91% ID/g) at 1 h post-administration with [²¹³Bi][Bi(neunpa-Ph-Pip-Nle-CycMSH_hex)]. This study demonstrates H₄neunpa-NH₂ to be an effective chelating ligand for [²¹³Bi]Bi³⁺ and [¹¹¹In]In³⁺, with promising characteristics for further development toward theranostic applications.