Diethylethanolamine → Deoxyuridine Triphosphate

HRID 230076

반응식

HRID 230076 반응식
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]

참가 물질

7

반응물 3

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

생성물 2

HCID 65070Deoxyuridine TriphosphateHCID 1385867Triethylammonium

시약·촉매·용매 2

HCID 962Water용매HCID 6228Dimethylformamide용매

실험 절차

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. 1HNMR [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). 31P [D2O]: −9.91 (γP, d, J 19.2), [−11.08(αP, d, J20.1) and −11.30 (αP, d, J 20.1), due to two diastereoisomers] and −22.57 (βP, m). MS[(ES(−)], m/z 1369.1 (M−).

후처리

  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

출처 및 근거

ORD 기록
ord-6a86064c14e54ad9983a362b9d1e9d40
출처 데이터셋
uspto-grants-2009_07
특허 번호
US07566537B2

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