The meta-vinylhalide fluoroalkyl ester nortropanes 1-4 were synthesized as ligands of the serotonin transporter (SERT) for use as positron emission tomography (PET) imaging agents. In vitro competition binding assays demonstrated that 1-4 have a high affinity for the SERT (Ki's = 0.3 - 0.4 nM) and are selective for the SERT over the dopamine and norepinephrine transporters (DAT and NET). MicroPET imaging in anesthetized cynomolgus monkeys with [18F]1-[18F]4 demonstrated that all four tracers behave similarly with peak uptake in the SERT-rich brain regions achieved after 45-55 min followed by a steady washout. An awake monkey study was performed with [18F]1 which demonstrated that the uptake of [18F]1 was not influenced by anesthesia. Chase studies with the SERT ligand 15 displaced [18F]1-[18F]4 but chase studies with the DAT ligand 16 did not displace [18F]1-[18F]4 thus indicating that the tracers were binding specifically to the SERT.
The non-natural amino acids (R)- and (S)-2-amino-3-fluoro-2-methylpropanoic acid 5 and (R)- and (S)-3-fluoro-2-methyl-2-N-(methylamino)propanoic acid 8 were synthesized in shorter reaction sequences than in the original report starting from enantiomerically pure (S)- and (R)-α-methyl-serine, respectively. The reaction sequence provided the cyclic sulfamidate precursors for radiosynthesis of (R)- and (S)-[18F]5 and (R)- and (S)-[18F]8 in fewer steps than in the original report. (R)- and (S)-[18F]5 and(R)- and (S)-[18F]8 were synthesized by no-carrier-added nucleophilic [18F]fluorination in 52–66% decay-corrected-yields with radiochemical purity over 99%. The cell assays showed that all four compounds were substrates for amino acid transport and enter 9L rat gliosarcoma cells in vitro at least in part by system-A amino acid transport. The biodistribution studies demonstrated that in vivo tumor to normal brain ratios for all compounds were high with ratios of 20:1 to115:1 in rats with intracranial 9L tumors. The (R)- enantiomers of [18F]5 and [18F]8 demonstrated higher tumor uptake in vivo compared to the (S)- enantiomers.
99mTc-mercaptoacetyltriglycine (99mTc-MAG3), 99mTc-dd- and ll-ethylene-di-cysteine (99mTc-EC) and 99mTc-mercaptoacetamide-ethylene-cysteine (99mTc-MAEC) contain N3S or N2S2 ligands designed to accommodate the four ligating sites of the {99mTcO}3+ core; they are all excellent renal imaging agents but have renal clearances less than that of 131I-orthoiodohippurate (131I-OIH). To explore the potential of the newly accessible but less polar {99mTc(CO)3}+ core having three ligating sites, we decided to build on the success of 99mTc-EC with its N2S2 ligand and two dangling carboxylates and have chosen an N2S ligand that also has two dangling carboxyls, lanthionine (LANH2), to form 99mTc(CO)3(LAN), a new renal radiopharmaceutical.Methods
Biodistribution studies were performed on Sprague-Dawley rats by using 99mTc(CO)3(LAN) isomers, meso-LAN and dd,ll-LAN (an enantiomeric mixture), coinjected with 131I-OIH. Human studies were also performed by coinjecting each 99mTc product (~74 MBq [~2 mCi]) and 131I-OIH (~ 7.4 MBq [~ 0.2 mCi]) into 3 normal volunteers with dual-isotope imaging performed by using a camera system fitted with a high-energy collimator. Blood samples were obtained from 3 to 90 min after injection, and urine samples were obtained at 30, 90 and 180 min.
Results
Biodistribution studies in rats revealed a rapid blood clearance as well as rapid renal extraction for both preparations, with the dose in urine at 60 min averaging 88% that of 131I-OIH. In humans, both agents provided excellent renal images, with the plasma clearance averaging 228 mL/min for 99mTc(CO)3(meso-LAN) and 176 mL/min for 99mTc(CO)3(dd,ll-LAN), respectively. At 3 hours, both 99mTc(CO)3(meso-LAN) and 99mTc(CO)3(dd,ll-LAN), showed good renal excretion, averaging 85% and 77% that of 131I-OIH, respectively. Plasma protein binding was minimal (10% and 2%), and red cell uptake was similar (24% and 21%) for 99mTc(CO)3(meso-LAN) and 99mTc(CO)3(dd,ll-LAN), respectively.
Conclusion
Although the plasma clearance and the rate of renal excretion of the 99mTc(CO)3(LAN) complexes are still less than those of 131I-OIH, the results of this first application of a 99mTc tricarbonyl complex as a renal radiopharmaceutical in humans demonstrate that 99mTc(CO)3(LAN) complexes are excellent renal imaging agents and support continued renal radiopharmaceutical development based on the 99mTc tricarbonyl core.