Diethylethanolamine → Deoxyuridine Triphosphate

HRID 230076

Reaktionsgleichung

Reaktionsgleichung von HRID 230076
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]

Komponenten

7

Edukt 3

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

Produkt 2

HCID 65070Deoxyuridine TriphosphateHCID 1385867Triethylammonium

Reagenzien, Katalysatoren, Lösungsmittel 2

HCID 962WaterLösungsmittelHCID 6228DimethylformamideLösungsmittel

Versuchsvorschrift

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−).

Aufarbeitung

  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

Quellen und Nachweise

ORD-Eintrag
ord-6a86064c14e54ad9983a362b9d1e9d40
Quelldatensatz
uspto-grants-2009_07
Patentnummer
US07566537B2

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