20.500.12556/RUL-184316 Mitochondria-targeting moieties based on N-tethered pyridinium cations Mitochondria-targeting moieties (MTMs) are molecular fragments designed to deliver covalently tethered functional cargo to mitochondria, providing a modular strategy for chemical biology tools, imaging agents, and mitochondria-targeted therapies. Phosphonium- or nitrogen cation-based MTMs are not inert vectors and exhibit intrinsic bioactivity on mitochondrial and cellular levels to various extents. Here, we systematically evaluated a panel of N+-based cations to determine how structural features influence subcellular distribution and inherent bioactivity. Live-cell imaging of fluorescent dye conjugates revealed that 3,5-diphenylpyridinium (DPPy+) exhibits cellular uptake and mitochondrial targeting comparable to the benchmark triphenylphosphonium (TPP+), whereas conjugates with unsubstituted pyridinium preferentially accumulate in lysosomes. Profiling of inert cargo derivatives showed that DPPy+ has lower intrinsic activity on mitochondrial membrane potential and oxidative phosphorylation, as well as on cellular respiration and viability than TPP+. The combination of efficient mitochondrial delivery and low intrinsic bioactivity translated to bioactive cargo: a Kv1.3 inhibitor conjugate with DPPy+ induced apoptosis in cancer cell lines and demonstrated improved cancer selectivity relative to the TPP+ conjugate in pancreatic organoid models. These results position lipophilic pyridinium cations as effective TPP+ surrogates with enhanced biocompatibility for mitochondria-targeted therapeutic and diagnostic agents, while revealing the structure-dependent competing lysosomal accumulation of permanent nitrogen cations. cancer cations fluorescentprobes medicinal chemistry mitochondria rak kationi fluorescentne sonde medicinska kemija mitohondriji true false true Angleški jezik Slovenski jezik Članek v reviji 2026-07-03 16:38:57 2026-07-03 16:39:07 2026-07-29 13:36:53 0000-00-00 00:00:00 2026 0 0 13 str. iss. 29 , art. e7158257 Vol. 65 2026 0000-00-00 Zaloznikova Objavljeno NiDoloceno 0000-00-00 0000-00-00 0000-00-00 615.4:54:616-006 1521-3773 10.1002/anie.7158257 279001347 RAZ_Dzajic_Ivan_2026.pdf RAZ_Dzajic_Ivan_2026.pdf 1 7F94C1804BD9E71E43918A01BF954840 77e00068d1923dee70f55589f745f8a607b58ddc369abd58de00d25a87a579d3 cd79dc56-80cf-11f1-9b0d-0050569b8976 https://repozitorij.uni-lj.si/Dokument.php?lang=slv&id=240134 https://onlinelibrary.wiley.com/doi/10.1002/anie.7158257 1 d856c247-76ec-11f1-9b0d-0050569b8976 https://repozitorij.uni-lj.si/Dokument.php?lang=slv&id=236990 Medicinska fakulteta Fakulteta za farmacijo Biotehniška fakulteta 0 0 0