Water → 2'-(N-Lysyl)amino-2'-deoxy-cytidine Triphosphate

HRID 1018820Notes

Reaction equation

HRID 1018820 reaction equation
[NH2:1][C@H:2]([C:8]([NH:10][C@@H:11]1[C@H:15]([OH:16])[C@@H:14]([CH2:17][OH:18])[O:13][C@H:12]1[N:19]1[CH:26]=[CH:25][C:23]([NH2:24])=[N:22][C:20]1=[O:21])=[O:9])[CH2:3][CH2:4][CH2:5][CH2:6][NH2:7].[P:27](Cl)(Cl)(Cl)=[O:28].[O-:32][P:33]([O:36][P:37]([O-:40])([O-:39])=[O:38])(=O)[O-:34].C([NH+](CCCC)CCCC)CCC.C([NH+](CCCC)CCCC)CCC.C([NH+](CCCC)CCCC)CCC.C([NH+](CCCC)CCCC)CCC.[O-]P(OP([O-])([O-])=O)(=O)[O-].CN.[NH4+].[OH-:105].[O-]P(OP(OP([O-])([O-])=O)([O-])=O)(=O)[O-].[OH2:119]>P(OCC)(OCC)(OCC)=O.C(#N)C.CC(O)C>[P:33]([O:18][CH2:17][C@H:14]1[O:13][C@@H:12]([N:19]2[CH:26]=[CH:25][C:23]([NH2:24])=[N:22][C:20]2=[O:21])[C@H:11]([NH:10][C:8](=[O:9])[C@H:2]([CH2:3][CH2:4][CH2:5][CH2:6][NH2:7])[NH2:1])[C@@H:15]1[OH:16])([O:36][P:37]([O:40][P:27]([OH:28])([OH:119])=[O:105])([OH:39])=[O:38])(=[O:34])[OH:32]

Participants

12

Reactant 8

HCID 962WaterHCID 6329MethylamineHCID 14923Ammonium HydroxideHCID 24813Phosphorus oxychlorideHCID 644102PyrophosphateHCID 3440921TriphosphateHCID 21910867Tributyl ammonium pyrophosphateHCID 228991812'-(N-lysyl)amino 2'-deoxy cytidine

Product 1

HCID 228991862'-(N-Lysyl)amino-2'-deoxy-cytidine Triphosphate

Reagents, catalysts & solvents 3

HCID 3776IsopropanolSolventHCID 6342AcetonitrileSolvent2×HCID 6535Triethyl PhosphateSolvent

Conditions

Temperature
0 °C

Procedure

2′-(N-lysyl)amino 2′-deoxy cytidine (0.180 g, 0.22 mmol) was dissolved in triethyl phosphate (2.00 ml) under Ar. The solution was cooled to 0° C. in an ice bath. Phosphorus oxychloride (99.999%, 3 eq., 0.0672 mL) was added to the solution and the reaction was stirred for two hours at 0° C. Tributylammonium pyrophosphate (4 eq., 0.400 g) was dissolved in 3.42 mL of acetonitrile and tribuytylamine (0.165 mL). Acetonitrile (1 mL) was added to the monophosphate solution followed by the pyrophosphate solution which was added dropwise. The solution was clear. The reaction was allowed to warm up to room temperature. After stirring for 45 minutes, methyl amine (5 mL) was added and the reaction and stirred at room temperature for 2 hours. A two phase mixture appeared (little beads at the bottom of the flask). TLC (7:1:2 iPrOH:NH4OH:H2O) showed the appearance of a triphosphate material. The solution was concentrated, dissolved in water and loaded on a newly prepared DEAE Sephadex A-25 column. The column was washed with a gradiant up to 0.6 M TEAB buffer and the product eluted off in fractions 90-95. The fractions were analyzed by ion exchange HPLC. Each fraction showed one triphosphate peak that eluted at ˜4.000 minutes. The fractions were combined and pumped down from methanol to remove buffer salt to yield 15.7 mgs of product.

Workup

  1. additionwas added dropwise
  2. temperatureto warm up to room temperature
  3. stirringAfter stirring for 45 minutes
  4. customthe reaction
  5. stirringstirred at room temperature for 2 hours
  6. concentrationThe solution was concentrated
  7. dissolutiondissolved in water
  8. washThe column was washed with a gradiant up to 0.6 M TEAB buffer
  9. washthe product eluted off in fractions 90-95
  10. washthat eluted at ˜4.000 minutes
  11. customto remove buffer salt

Sources & evidence

ORD record
ord-b00396d83aa6400e9dba41e38b97947e
Source dataset
uspto-grants-2002_11
Patent number
US06482932B1

Reaction notes

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