H₃TPAN-Triazole-Bn-NH₂: Tripicolinate Clicked-Bifunctional Chelate for [²²⁵Ac]/[¹¹¹In] Theranostics

A new, high-denticity, bifunctional ligand─H₃TPAN-triazole-Bn-NH₂─has been synthesized and studied in complexation with [²²⁵Ac]Ac³⁺ and [¹¹¹In]In³⁺ for radiopharmaceutical applications. The bifunctional chelator is readily synthesized, using a high-yielding four-step prep, which is highly adaptable and allows for straightforward incorporation of different covalent linkers using Cu^I-catalyzed alkyne-azide cycloaddition (click) chemistry. Nuclear magnetic resonance (NMR) studies of H₃TPAN-triazole-Bn-NH₂ with La³⁺ and In³⁺ metal ions show the formation of a single, asymmetric complex with each ion in solution, corroborated by density functional theory (DFT) calculations. Radiolabeling studies with [²²⁵Ac]Ac³⁺ and [¹¹¹In]In³⁺ showed highly effective complexation, achieving quantitative radiochemical conversions at low ligand concentrations (<10^-6 M) under mild conditions (RT, 10 min), which is further accompanied by high stability in human serum. The bioconjugate─H₃TPAN-triazole-Bn-Aoc-Pip-Nle-CycMSH_hex─was prepared for targeting of MC1R-positive tumors, and the corresponding ¹¹¹In-radiolabeled tracer was studied in vivo. SPECT/CT and biodistribution studies in C57BL/6J mice bearing B16-F10 tumors were performed, with the radiotracer showing good in vivo stability; tumor uptake was achieved. This work highlights a new promising and versatile bifunctional chelator, easily prepared and encouraging for ²²⁵Ac/¹¹¹In theranostics.