反应详情

温度

50 °C

Uridine 5′-monophosphate (Sigma, Milwaukee, 3.0 g, 9.26 mmol) was dissolved in dry DMF (10 mL) and tributylamine (Aldrich, 2 mL). The solution was evaporated in vacuo at 40° C. to an oil. The residue was dissolved in dry DMF (Aldrich, 8 mL) to form a solution. Carbonyldilmidazole (Aldrich, 1.65 g, 10.18 mmol) was added to this solution. The reaction was heated at 50° C. for one hour. Uridine 5′-triphosphate (Yamasa, 5.60 g, 10.18 mmol) prepared as the anhydrous tributylammonium salt in DMF (5 mL) and tributylamine (2 mL), as described in Example 3 below, was added to the reaction solution. The mixture was allowed to stir at 50° C. for three days when the solution was evaporated in vacuo to an oil, redissolved in water (5 mL) and purified by column (300×50 mm) chromatography (Sephadex DEAE-A25, 40-120μ, Aldrich, pre-swollen in 1.0 M NaHCO3 and washed with 2 column volumes of deionized H2O (H2O→0.03 M NH4HCO3 gradient). The pure fractions were concentrated in vacuo at 35° C., and H2O added and reevaporated 5 times to obtain diuridine tetraphosphate tetraammonium salt as a white solid (2.37 g, 30% yield): 92.11% pure by HPLC with the same retention time as the standard. In addition, the tetraammonium salt was analyzed by FABMS to give a mass of [C18H25N4O23P4 (M−H+)−: calculated 788.9860] 788.9857, confirming a parent formula of C18H26N4O23P4 for the free acid].

1. customThe solution was evaporated in vacuo at 40° C. to an oil
2. dissolutionThe residue was dissolved in dry DMF (Aldrich, 8 mL)
3. customto form a solution
4. additionCarbonyldilmidazole (Aldrich, 1.65 g, 10.18 mmol) was added to this solution
5. additionwas added to the reaction solution
6. customwas evaporated in vacuo to an oil
7. dissolutionredissolved in water (5 mL)
8. custompurified by column (300×50 mm) chromatography (Sephadex DEAE-A25, 40-120μ, Aldrich
9. washpre-swollen in 1.0 M NaHCO3 and washed with 2 column volumes of deionized H2O (H2O→0.03 M NH4HCO3 gradient)
10. concentrationThe pure fractions were concentrated in vacuo at 35° C.
11. additionH2O added
12. customreevaporated 5 times