Inorganic Chemistry, Vol.59, No.7, 4895-4908, 2020
H(2)CHXhox: Rigid Cyclohexane-Reinforced Nonmacrocyclic Chelating Ligand for [Ga-nat/67/68]Ga3+
A rigid chiral acyclic chelator H(2)CHXhox was synthesized and evaluated for Ga3+-based radiopharmaceutical applications; it was compared to the previously reported hexadentate H-2 hox to determine the effect of a backbone reinforced from adding a chiral 1S,2S-trans-cyclohexane on metal complex stability, kinetic inertness, and in vivo pharmacokinetics. NMR spectroscopy and theoretical calculation revealed that [Ga(CHXhox)](+) showed a very similar coordination geometry to that of [Ga(hox)](+), and only one isomer in solution was observed by NMR spectroscopy. Solution studies showed that the modification results in a significant improvement in the exceptionally high thermodynamic stability of [Ga(hox)](+) with a 1.56 log unit increase in stability constant (logK(ML) = 35.91(1)). More importantly, H(2)CHXhox showed very fast Ga3+ complexation at physiological pH 7.4, and acid-assisted Ga3+ complex dissociation kinetic studies (pH 1) in comparison with H(2)hox revealed a 50-fold increase of the dissociation half-life time from 73 min to 58 h. Fluorescence microscopy imaging study confirmed its cellular uptake and accumulation in endoplasmic reticulum and mitochondria. MTT studies indicated a quite low cytotoxicity of [Ga(CHXhox)](+) over a large concentration range. Dynamic PET imaging studies showed no accumulation in muscle, lungs, bone, and brain, suggesting no release of free Ga3+ ions. [Ga-68][Ga(CHXhox)](+) is cleared from the mouse via hepatobiliary and renal pathways. Compared to [Ga-68][Ga(hox)](+), the increased lipophilicity of [Ga-68][Ga(CHXhox)](+) enhanced heart and liver uptake and decreased kidney clearance. [Ga-67][Ga(CHXhox)](+) SPECT/CT imaging and biodistribution study revealed good clearance from liver to gallbladder after 90 min and finally into feces after 5 h. No decomposition or transchelation was observed over the 5 h study. These results confirmed H(2)CHXhox to be an obvious improvement over H(2)hox and an excellent candidate in this new "ox" family for the development of radiopharmaceutical compounds.