Trimethyl Phosphate → 5-(4-(N-trifluoroacetyl-2-aminoethoxy)but-2-yn-1-yl)-2',3'-dideoxyuridine-5'-monophosphate

반응식

[F:1][C:2]([F:29])([F:28])[C:3]([NH:5][CH2:6][CH2:7][O:8][CH2:9][C:10]#[C:11][CH2:12][C:13]1[C:14](=[O:27])[NH:15][C:16](=[O:26])[N:17]([CH:25]=1)[C@@H:18]1[O:24][C@H:21]([CH2:22][OH:23])[CH2:20][CH2:19]1)=[O:4].O=P(Cl)(Cl)Cl.[P:35](OC)([O:39]C)([O:37]C)=[O:36]>>[P:35]([O:23][CH2:22][C@H:21]1[O:24][C@@H:18]([N:17]2[CH:25]=[C:13]([CH2:12][C:11]#[C:10][CH2:9][O:8][CH2:7][CH2:6][NH:5][C:3](=[O:4])[C:2]([F:28])([F:1])[F:29])[C:14](=[O:27])[NH:15][C:16]2=[O:26])[CH2:19][CH2:20]1)([OH:39])([OH:37])=[O:36]

참가 물질

반응물 3

HCID 10541Trimethyl Phosphate

HCID 24813Phosphorus oxychloride

HCID 600251125-(4-(n-Trifluoroacetyl-2-aminoethoxy)but-2-yn-1-yl)-2',3'dideoxyuridine

생성물 1

HCID 600251115-(4-(N-trifluoroacetyl-2-aminoethoxy)but-2-yn-1-yl)-2',3'-dideoxyuridine-5'-monophosphate

실험 절차

To a solution of 5 (30 mg, 0.0738 mmol) in P(O)(OCH3)3 (0.5 mL) was treated with POCl3 (70 μL, 0.73 mmol) at 0° C., followed by stirring at r.t for 2-3 h and then evaporating solvent under reduced pressure and purification by preparative reverse phase HPLC to afford the corresponding mono phosphate 6. 31P NMR (CD3OD, 121.4 MHz) δ 0.79 (s); 19F NMR (CDCl3), 282.2 MHz) δ −77.37 (s, CF3). To a dried solution of 6 in DMF (1 mL) were added 1,1′-carbonyldiimidazole (CDI, 33 mg, 0.21 mmol), followed by stirring at r.t. for 5 h and then quenching with MeOH (8 μL, 0.21 mmol). A solution of tetrabutylammonium pyrophosphate (TBAPP, 143.0 mg, 0.33 mmol) and tributyl amine (0.15 mL, 0.66 mmol) in DMF (1.5 mL) was added to the above solution at r.t. under Ar, followed by stirring overnight and then quenched with 2.0 M TEAB. After evaporating solvent, the organic residue was purified by preparative reverse phase HPLC with 100 mM TEAA/AcCN (0 to 50% gradient) to give the corresponding triphosphate 7. 1H NMR (D2O, 300 MHz) δ 7.74 (s, 1H, H6), 5.95 (dd, 1H, H1′, J=3.3 and 6.3 Hz), 4.29-4.20 (m, 1H, H4′), 4.18-4.07 (m, 3H, CH2 and H5′), 4.02-3.92 (m, 1H, H5′), 3.58 (t, 2H, CH2, J=5.1 Hz), 3.38 (t, 2H, CH2, J=5.1 Hz), 3.20 (s, 2H, CH2), 2.35-2.22 (m, 1H, H2), 2.10-1.92 (m, 3H, H2, and 2H3′). 31P NMR (D2O, 121.4 MHz) δ −10.52 (d, J=19.2 Hz), −10.95 (d, J=20.3 Hz), −22.96 (t, J=20.3 Hz); 19F NMR (D2O, 282.2 MHz) δ −76.37 (s, CF3). Removal of trifluoro acetate (TFA) protecting group was accomplished by treatment with NH4OH (33%, 500 μL) at 60° C. for 20 min and then at r.t for 1 h to give the corresponding triphosphate 8 (17 mM, 500 μL).