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49949094097
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note
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Chemical synthesis and characterization data.(a) General methods.All chemicals, solvents, and reagents were purchased from Sigma-Aldrich (St. Louis, MO) and used directly without further purification unless otherwise noted. 1,4,7,10-Tetraazacyclododecane-1,4,7-tris-(t-butyl acetate) (DO3A-t-Bu-ester) was obtained from Macrocylic Inc. (Dallas, TX). Silica gel 60 (70-230 mesh) used for column chromatography was obtained from Sigma-Aldrich. Analytical thin-layer chromatography (TLC) was performed using Merck 60 F254 silica gel (precoated sheets, 0.2 mm thick) (Lawrence, KS) and Whatman MCK 18F reversed-phase plates (Maidstone, Kent, UK).
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49949088125
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note
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111In-DO3A-BP was determined by adding 5 μL of the complex to a solution containing 500 μL of Milli-Q water and 500 μL of octanol (obtained from a saturated octanol/water solution) (n = 10). The sample vials were shaken for 1 h at room temperature. From each vial, aliquots of 100 μL each were removed from the octanol phase and the water phase, respectively, and counted separately. The partition coefficient was calculated as the ratio of counts in the octanol fraction to the counts in the water fraction. An average of log P value was obtained from the 10 samples.
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49949093597
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note
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111In-DO3A-BP was measured by counting the cell lysates in a γ-counter. The protein concentration in each lysate was determined using the Bio-Rad Protein Assay. The assay was repeated 5 times, and all data were pooled for analysis.
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33
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49949091992
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note
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111In-DO3A-BP was diluted with PBS (10 mM) to prepare the injection doses. Each mouse was injected with ca. 5 μCi of radioactivity in 100 μL via the tail vein. The mice were anesthetized prior to sacrifice at each time point (1, 4, 24, and 48 h; n = 4 per time point). Organs of interest were removed, weighed, and counted by a γ-counter. Standards were prepared and counted along with the samples to calculate the percent injected dose per gram (%ID/g) and percent injected dose per organ (%ID/organ). The animals of the last time point groups were housed in metabolic cages (4 mice per cage) to collect urine and feces at 1, 4, 24, and 48 h pi to evaluate the excretion route and stability of the compound.
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