Radiosynthesis, in vitro cellular uptake and in vivo biodistribution of 3′- O-(3-benzenesulfonylfuroxan-4-yl)-5-[ 125I]iodo-2′-deoxyuridine, a nucleoside-based nitric oxide donor

3′- O-(3-Benzenesulfonylfuroxan-4-yl)-5-iodo-2′-deoxyuridine ( 1) is a cytotoxic nitric oxide (NO) donor-nucleoside dual action prodrug designed to exploit both NO and an antimetabolite nucleoside for cancer therapy. 1 was radiolabeled by radioiodide exchange and purified by HPLC in 16% overall radiochemical yield. The specific activity of [ 125I] 1 was 31.8 μCi/μg (680 MBq/μM). Protein binding, deiodination, cellular uptake and incorporation of 1 into cellular nucleic acids were measured by standard methods, and its in vivo biodistribution was determined in Balb/c mice bearing implanted EMT-6 tumors following intravenous injection. [ 125I] 1 degraded rapidly during the in vitro tests, thus impeding unequivocal assessment but indicating that it was only weakly protein bound and that it was resistant to deiodination under test conditions. Uptake of [ 125I] 1 by murine tumor cells (KBALB and KBALB-STK) in vitro was low (∼17 fmol/μg protein over 2 h) with only ∼0.3% (0.04–0.06 fmol/μg protein) of total uptake present in the DNA fraction. In the murine tumor model, liver, kidney, intestine and tumor showed the greatest uptake, with liver, intestine and blood all containing >5 injected dose per gram of tissue (%ID/g) during the 15-min to 2-h postinjection period. Maximum tissue/blood level ratios were 3.6 (2 h) for tumor and 6.4 (24 h) for liver. Low uptake in thyroid and stomach was indicative of minimal in vivo deiodination. [ 125I] 1 undergoes only minimal deoiodination under both in vitro and in vivo conditions. Under conditions of the in vitro NO release assay, 1 reacts to produce a single, major, unstable adduct that decomposes upon workup. Protein binding of [ 125I] 1 could not be assessed because of similar chemical reaction with albumin. Incorporation of radioactivity into the cellular nucleic acid fraction was low, and in vivo distribution of [ 125I] 1 was consistent with nonspecific reactivity towards tissue nucleophiles. The chemical reactivity of [ 125I] 1 mitigates against its use as a NO donor and as a tracer for this class of compounds.