Diethylethanolamine → Deoxyuridine Triphosphate

HRID 1052870

反応式

HRID 1052870 の反応式
C(=O)(ON1C(=O)CCC1=O)ON1C(=O)CCC1=O.[P:19]([O:31][CH2:32][C@H:33]1[O:37][C@@H:36]([N:38]2[CH:45]=[C:44](C#CCN)[C:42](=[O:43])[NH:41][C:39]2=[O:40])[CH2:35][C@@H:34]1[OH:50])([O:22][P:23]([O:26][P:27]([OH:30])([OH:29])=[O:28])([OH:25])=[O:24])(=[O:21])[OH:20].[CH3:51][CH2:52][N:53]([CH2:56][CH2:57]O)[CH2:54][CH3:55]>CN(C=O)C.O>[CH:44]1[C:42](=[O:43])[NH:41][C:39](=[O:40])[N:38]([C@@H:36]2[O:37][C@H:33]([CH2:32][O:31][P:19]([O:22][P:23]([O:26][P:27]([OH:30])([OH:29])=[O:28])([OH:25])=[O:24])([OH:21])=[O:20])[C@@H:34]([OH:50])[CH2:35]2)[CH:45]=1.[CH2:52]([NH+:53]([CH2:56][CH3:57])[CH2:54][CH3:55])[CH3:51]

参加物

7

反応物 3

HCID 7497DiethylethanolamineHCID 676246Di(succinimido) carbonateHCID 18664765ap-dUTP

生成物 2

HCID 65070Deoxyuridine TriphosphateHCID 1385867Triethylammonium

試薬・触媒・溶媒 2

HCID 962Water溶媒HCID 6228Dimethylformamide溶媒

実験手順

TMR-sieber linker free acid (4.34 mg, 5 μmol) was stirred with disuccinimidyl carbonate (1.74 mg, 7.5 μmol) and N,N-dimethyl aminopyridine (0.92 mg, 7.5 μmol) in DMF (1 mL) at room temperature. After 10 minutes, all the reaction mixture was added to tetra-(tri-butylammonium) salt of 5-(3-aminopropynyl)-2′-deoxyuridine-5′-triphosphate (10 μmol). The reaction was stirred at room temperature for 4 hrs and stored in the fridge overnight. The reaction mixture was then diluted with chilled water (10 mL) and all the resulting solution was applied onto a short column of DEAE A-25. The column was initially eluted with 0.1 M TEAB buffer and then 0.7 M TEAB buffer. The 0.7 M TEAB eluents were collected and evaporated under reduced pressure. The residue was co-evaporated with MeOH (2×10 mL) and then purified by preparative HPLC. The title compound was obtained as triethylammonium salt in 31% yield (based on the quantification of TMR at 555 nm in water (pH 7)). 1HNMR in D2O indicated two diastereoisomers, due to the sieber linker moiety and there were approximately three triethylammonium count ions. 1H NMR [D2O]: 8.18 (1H, m), 8.06 (1H, m), 7.76 (0.55H, s), 7.74 (0.45H, s), 7.36-7.09 (5H, m), 6.89-6.72 (3H, m), 6.59-6.37 (5H, m), 6.12 (0.55H, t, J 6.6), 6.05 (0.45H, t, J 6.6), 5.99 (0.45H, d, J 2.5), 5.91 (1.1H, m), 5.88 (0.45H, s), 4.49 (0.55H, m), 4.43 (0.45H, m), 4.00-3.35 (9H, m), 3.30-2.95 (32H, m), 2.65-2.52 (4H, m), 2.25-2.05 (4H, m), 1.62-1.42 (4H, m) and 1.23 (27H, t, J 7.3). −P [D2O]: −9.91 (γP, d, J 19.2), [−11.08 (αP, d, J 20.1) and −11.30 (αP, d, J 20.1), due to two diastereoisomers] and −22.57 (βP, m). MS[(ES(−)], m/z 1369.1 (M−).

後処理

  1. customstored in the fridge overnight
  2. washThe column was initially eluted with 0.1 M TEAB buffer
  3. customThe 0.7 M TEAB eluents were collected
  4. customevaporated under reduced pressure
  5. custompurified by preparative HPLC

出典と根拠

ORDレコード
ord-7e2b840d8ca94d0c87c0d809659bdbc6
出典データセット
uspto-grants-2016_07
特許番号
US09388463B2

関連ノート

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