Uridine → rUrd.rUrd.P-P-P-P

HRID 1266538Notes

Reaction equation

HRID 1266538 reaction equation
[C@@H:1]1([N:10]2[CH:17]=[CH:16][C:14](=[O:15])[NH:13][C:11]2=[O:12])[O:9][C@H:6]([CH2:7][OH:8])[C@@H:4]([OH:5])[C@H:2]1[OH:3].C(N1C=CN=C1)(N1C=CN=C1)=O.[P:30]([O:42][CH2:43][C@H:44]1[O:48][C@@H:47]([N:49]2[CH:56]=[CH:55][C:53](=[O:54])[NH:52][C:50]2=[O:51])[C@H:46]([OH:57])[C@@H:45]1[OH:58])([O:33][P:34]([O:37][P:38]([OH:41])([OH:40])=[O:39])([OH:36])=[O:35])(=[O:32])[OH:31].C([NH+](CCCC)CCCC)CCC>CN(C=O)C.C(N(CCCC)CCCC)CCC>[OH:40][P:38]([O:37][P:34]([O:33][P:30]([O:42][P:30]([OH:33])([OH:32])=[O:31])([OH:31])=[O:32])([OH:36])=[O:35])(=[O:39])[OH:41].[C@@H:1]1([N:10]2[CH:17]=[CH:16][C:14](=[O:15])[NH:13][C:11]2=[O:12])[O:9][C@H:6]([CH2:7][OH:8])[C@@H:4]([OH:5])[C@H:2]1[OH:3].[C@@H:47]1([N:49]2[CH:56]=[CH:55][C:53](=[O:54])[NH:52][C:50]2=[O:51])[O:48][C@H:44]([CH2:43][OH:42])[C@@H:45]([OH:58])[C@H:46]1[OH:57]

Participants

7

Reactant 4

HCID 6029UridineHCID 6133Uridine 5'-triphosphateHCID 682631,1'-CarbonyldiimidazoleHCID 3724478Tributylammonium

Product 1

HCID 131723853rUrd.rUrd.P-P-P-P

Reagents, catalysts & solvents 2

HCID 6228DimethylformamideSolvent2×HCID 7622TributylamineSolvent2×

Conditions

Temperature
50 °C

Procedure

Uridine 5′-monrophosphate (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. Carbonyldiimidazole (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→0.3 M NH4HCO3 gradient). The pure fractions were concentrated in vacuo at 35° C., and H2O added and reevaporated 5 times to obtain diuridine tetraphosphate tetrammonium 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 tetrammonium 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].

Workup

  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. additionwas added to the reaction solution
  5. customwas evaporated in vacuo to an oil
  6. dissolutionredissolved in water (5 mL)
  7. custompurified 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→0.3 M NH4HCO3 gradient)
  8. concentrationThe pure fractions were concentrated in vacuo at 35° C.
  9. additionH2O added
  10. customreevaporated 5 times

Sources & evidence

ORD record
ord-f959edc9103745df9b7640f15fb7f7ba
Source dataset
uspto-grants-2003_04
Patent number
US06548658B2

Reaction notes

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