Iodine → ap-dUTP

HRID 1168688

反応式

HRID 1168688 の反応式
C([O:4][C@@H:5]1[C@@H:9]([CH2:10][OH:11])[O:8][C@@H:7]([N:12]2[CH:19]=[C:18]([C:20]#[C:21][CH2:22][NH:23]C(=O)C(F)(F)F)[C:16](=[O:17])[NH:15][C:13]2=[O:14])[CH2:6]1)(=O)C.Cl[P:31]1[O:36]C(=O)C2C=CC=CC=2[O:32]1.[O-:42][P:43]([O:46][P:47]([O-])([O-:49])=[O:48])(=[O:45])[O-:44].C([NH+](CCCC)CCCC)CCC.C([NH+](CCCC)CCCC)CCC.C([NH+](CCCC)CCCC)CCC.C([NH+](CCCC)CCCC)CCC.C(N(CCCC)CCCC)CCC.II.[O-:118]S([O-])=O.[Na+].[Na+]>N1C=CC=CC=1.O1CCOCC1.CN(C=O)C.N1C=CC=CC=1.O>[P:31]([O:11][CH2:10][C@H:9]1[O:8][C@@H:7]([N:12]2[CH:19]=[C:18]([C:20]#[C:21][CH2:22][NH2:23])[C:16](=[O:17])[NH:15][C:13]2=[O:14])[CH2:6][C@@H:5]1[OH:4])([O:36][P:47]([O:46][P:43]([OH:45])([OH:44])=[O:42])([OH:49])=[O:48])(=[O:118])[OH:32]

参加物

11

反応物 6

HCID 807IodineHCID 7622TributylamineHCID 24437Sodium SulfiteHCID 2370102-Chloro-4H-1,3,2-benzodioxaphosphorin-4-oneHCID 213622903'-O-Acetyl-5-(3-trifluoroacetamidopropyn-1-yl)-2'-deoxyuridineHCID 21910867Tributyl ammonium pyrophosphate

生成物 1

HCID 18664765ap-dUTP

試薬・触媒・溶媒 4

HCID 1049Pyridine溶媒2 回HCID 6228Dimethylformamide溶媒HCID 312751,4-Dioxane溶媒HCID 66926435Pyridine water溶媒

実験手順

Triphosphates were synthesized by the method of Ludwig and Eckstein (J. Ludwig and F. Eckstein (1989) J. Org. Chem. 54, 631). 3′-O-Acetyl-5-(3-trifluoroacetamidopropyn-1-yl)-2′-deoxyuridine (0.1 mmol, 41.9 mg) was coevaporated with pyridine and dissolved in dry pyridine (0.1 mL)/dry 1,4-dioxane (0.3 mL). 2-Chloro-4H-1,3,2-benzodioxaphosphorin-4-one (1.1 equiv., 0.11 mL 1 M solution) was added. A precipitate was formed and the mixture was stirred for 10 min. Then, tributylammonium pyrophosphate (70 mg) in dry DMF (0.35 mL)/tributylamine (0.1 mL) was added, giving a clear solution which was stirred for 10 min. An 1% solution of iodine in pyridine/water 98:2 (2.2 mL) was added and the immediately decolorized solution was stirred for another 15 min. After addition of a few drops of a 5% Na2SO3 solution it was evaporated at room temperature, the residue taken up in water (5 mL) and stirred at room temperature for 30 min. Then, concentrated aqueous ammonia (20 mL) was added and the mixture stirred at 50° C. for 4 h. The organic solvents were removed by rotary evaporation at room temperature and the residue purified by column chromatography (Sephadex DEAE cellulose, previously equilibrated with TEAB buffer) using a 0.2 M-0.6 M gradient of triethylammonium bicarbonate (TEAB) buffer solution. The UV active fractions were evaporated at room temperature to yield 5-(3-aminopropyn-1-yl)-2′-deoxyuridine triphosphate, contaminated with some diphosphate.

後処理

  1. customA precipitate was formed
  2. customgiving a clear solution which
  3. stirringwas stirred for 10 min
  4. stirringthe immediately decolorized solution was stirred for another 15 min
  5. customwas evaporated at room temperature
  6. stirringstirred at room temperature for 30 min
  7. additionThen, concentrated aqueous ammonia (20 mL) was added
  8. stirringthe mixture stirred at 50° C. for 4 h
  9. customThe organic solvents were removed by rotary evaporation at room temperature
  10. customthe residue purified by column chromatography (Sephadex DEAE cellulose
  11. customThe UV active fractions were evaporated at room temperature

出典と根拠

ORDレコード
ord-b5ac65873cc64cab9e0ffa4745966171
出典データセット
uspto-grants-2003_09
特許番号
US06627456B1

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