Diethylethanolamine → Deoxyuridine Triphosphate

HRID 1943539Notes

Reaction equation

HRID 1943539 reaction equation
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]

Participants

7

Reactant 3

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

Product 2

HCID 65070Deoxyuridine TriphosphateHCID 1385867Triethylammonium

Reagents, catalysts & solvents 2

HCID 962WaterSolventHCID 6228DimethylformamideSolvent

Procedure

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, J 20.1) and −11.30 (αP, d, J 20.1), due to two diastereoisomers] and −22.57 (βP, m). MS[(ES(−)], ink 1369.1 (M−).

Workup

  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

Sources & evidence

ORD record
ord-935fbdc5a2a841bba86ab77d8cba3951
Source dataset
uspto-grants-2013_03
Patent number
US08394586B2

Reaction notes

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