6808
Y. Taniguchi et al. / Bioorg. Med. Chem. 17 (2009) 6803–6810
(125 MHz, CDCl3): 169.8, 164.0, 154.9, 140.1, 135.7, 135.6, 133.8,
133.2, 133.0, 130.5, 129.7, 129.5, 128.8, 128.6, 128.4, 127.7,
127.6, 125.6, 107.7, 92.7, 91.7, 86.6, 80.3, 72.9, 62.9, 48.5, 26.8,
20.7, 19.2, 7.4. FTIR (neat): 1746, 1600 cmꢀ1. HRMS (ESI) m/z: calcd
for C42H45N2O7Si (M+H)+ 717.2991, found 717.3032.
34.6. FTIR (neat): 3368, 1633 cmꢀ1. HRMS (ESI) m/z: calcd for
C19H25N4O4 (M+H)+ 373.1870, found 373.1898.
4.3.3. (10S,30R,40R,50R,70R)-2-(40-Hydroxy-30-hydroxymethyl-10-
phenyl-20,60-dioxabicyclo[3.3.0]oct-70-yl)-4-methyl-3-pyrazolin-
5-one (9a)
4.2.5. (10S,30R,40R,50R,70R)-N2-Benzoyl-1-{40-acetoxy-10-phenyl-
30-(t-butyldiphenylsilyloxymethyl)-20,60-dioxabicyclo[3.3.0]oct-
70-yl}-4-methyl-3-pyrazolin-5-one (8a)
(WNA-2a
MP) 1H NMR (400 MHz, CD3OD): d 7.60 (2H, d,
J = 8.2 Hz), 7.47 (1H, s), 7.37 (2H, t, J = 7.3 Hz), 7.27 (1H, t,
J = 7.3 Hz), 6.38 (1H, t, J = 6.6 Hz), 4.45 (1H, d, J = 4.3 Hz), 4.15–4.12
(1H, m), 3.87–3.84 (2H, m), 3.69 (1H, dd, J = 6.4, 11.9 Hz), 2.87 (1H,
d, J = 6.6 Hz), 1.86 (3H, s). 13C NMR (125 MHz, CD3OD): 162.5,
143.6, 131.0, 129.4, 128.5, 126.2, 103.6, 93.0, 90.5, 83.2, 73.1, 63.4,
48.3, 7.0. FTIR (neat): 3354, 1708, 1603, 1528 cmꢀ1. HRMS (ESI) m/
z: calcd for C17H21N2O5 (M+H)+ 333.1445, found 333.1473.
(WNA-1a
MP) 1H NMR (400 MHz, CDCl3): d 8.19 (2H, dd, J = 1.2,
7.9 Hz), 7.67–7.21 (19H, m), 6.17 (1H, t, J = 6.5 Hz), 4.98 (1H, dd,
J = 4.5, 9.1 Hz), 4.71 (1H, d, J = 4.2 Hz), 4.21 (1H, d, J = 9.1 Hz),
4.00 (1H, dd, J = 2.4, 11.8 Hz), 3.65 (1H, dd, J = 5.5 Hz, 14.9 Hz),
3.59 (1H, dd, J = 2.4, 11.8 Hz), 2.88 (1H, dd, J = 7.9, 14.9 Hz), 1.92
(3H, s), 1.90 (3H, s), 0.97 (9H, s). FTIR (neat): 1750, 1600,
1239 cmꢀ1
.
HRMS (ESI) m/z: calcd for C42H45N2O7Si (M+H)+
4.3.4. (10S,30R,40R,50R,70S)-2-(40-Hydroxy-30-hydroxymethyl-10-
phenyl-20,60-dioxabicyclo[3.3.0]oct-70-yl)-4-methyl-3-pyrazolin-
5-one (9b)
717.2991, found 717.3028.
4.2.6. (10S,30R,40R,50R,70S)-N2-Benzoyl-1-{40-acetoxy-10-phenyl-
30-(t-butyldiphenylsilyloxymethyl)-20,60-dioxabicyclo[3.3.0]oct-
70-yl}-4-methyl-3-pyrazolin-5-one (8b)
(WNA-2bMP) 1H NMR (400 MHz, CD3OD): d 7.73 (2H, d,
J = 7.6 Hz), 7.38 (1H, s), 7.34 (2H, t, J = 7.3 Hz), 7.24 (1H, t,
J = 7.3 Hz), 6.00 (1H, t, J = 6.8 Hz), 4.66 (1H, d, J = 3.6 Hz), 4.05–
4.00 (1H, m), 3.88 (1H, dd, J = 2.1, 11.9 Hz), 3.81 (1H, dd, J = 3.6,
10.7 Hz), 3.68 (1H, dd, J = 6.1, 10.7 Hz), 3.12 (1H, dd, J = 7.9,
14.0 Hz), 2.69 (1H, dd, J = 6.1, 14.0 Hz), 1.92 (3H, s). 13C NMR
(125 MHz, CD3OD): 162.7, 142.5, 131.9, 129.3, 128.4, 126.6,
102.9, 93.2, 90.0, 84.5, 73.6, 63.7, 49.1, 6.9. FTIR (neat): 3386,
1609, 1530 cmꢀ1. HRMS (ESI) m/z: calcd for C17H21N2O5 (M+H)+
333.1445, found 333.1420.
(WNA-1bMP) 1H NMR (400 MHz, CDCl3): d 8.21 (2H, dd, J = 1.2,
7.9 Hz), 7.70–7.21 (19H, m), 6.14 (1H, t, J = 6.7 Hz), 5.04 (1H, dd,
J = 3.6, 9.2 Hz), 4.98 (1H, d, J = 3.9 Hz), 4.20 (1H, dt, J = 3.3,
9.4 Hz), 3.93 (1H, dd, J = 3.0, 11.6 Hz), 3.68 (1H, dd, J = 3.6,
11.6 Hz), 3.39 (1H, dd, J = 7.0, 14.3 Hz), 2.87 (1H, dd, J = 6.7,
14.3 Hz), 1.92 (3H, s), 1.90 (3H, s), 0.97 (9H, s). FTIR (neat): 1751,
1600, 1240 cmꢀ1. HRMS (ESI) m/z: calcd for C42H45N2O7Si (M+H)+
717.2991, found 717.2979.
4.4. General procedure of synthesis for the WNA-AP
phosphoramidite
4.3. General procedure for deprotection reaction
A THF solution of the above compound and TBAF (1.0 M THF
solution, 2 equiv) was stirred for 3.5 h at room temperature. MeOH
and a 0.2 M aqueous NaOH solution (2 equiv) were then added to
this mixture at 0 °C. After stirring for 2.5 h at 0 °C, the reaction
was quenched with acetic acid and diluted with MeOH. The sol-
vents were removed under reduced pressure. The residue was
purified by flash silica gel column chromatography (CHCl3/
CH3OH = 10:1) to give the corresponding diol compound.
DMTrCl (3 equiv) was added to a solution of the diol compound
in pyridine. After stirring for 1 h, the reaction mixture was diluted
with EtOAc, and then washed with water and brine. The organic
layer was washed with water and brine, dried over Na2SO4, evap-
orated, and then the residue was purified by flash silica gel column
chromatography (CHCl3/CH3OH = 99:1 containing 0.5% pyridine) to
give the corresponding DMTr-protected WNA. iP2NEt (6 equiv) was
added to
a solution of the above DMTr-WNA and iPr2NP(-
Cl)OC2H4CN (3 equiv) in CH2Cl2 at 0 °C. After stirring for 1 h, the
reaction mixture was diluted with an aqueous saturated NaHCO3
solution, and extracted with EtOAc. The organic layer was dried
over Na2SO4, evaporated, then the residue was purified by flash sil-
ica gel column chromatography (hexane/EtOAc = 1:1) to give the
purified materials.
4.3.1. (10S,30R,40R,50R,70R)-N3-Dimethylaminomethylidine-1-(40-
hydroxy-30-hydroxymethyl-10-phenyl-20,60-
dioxabicyclo[3.3.0]oct-70-yl)-3-aminopyrazole (4a)
(WNA-a
AP) 1H NMR (400 MHz, CDCl3): d 7.73 (1H, s), 7.50 (2H,
d, J = 7.3 Hz), 7.44 (1H, s), 7.38 (2H, t, J = 7.6 Hz), 7.28 (1H, t,
J = 7.3 Hz), 6.63 (1H, t, J = 7.1 Hz), 5.71 (1H, s), 4.53 (1H, d,
J = 6.7 Hz), 4.34–4.32 (1H, m), 4.04 (1H, d, J = 7.3 Hz), 4.00 (1H,
dd, J = 1.8, 10.8 Hz), 3.80 (1H, dd, J = 4.6, 10.8 Hz), 3.23 (1H, dd,
J = 6.7, 14.6 Hz), 3.03 (6H, s), 2.79 (1H, dd, J = 7.3, 14.6 Hz). 13C
NMR (125 MHz, CDCl3): 154.9, 151.7, 144.0, 140.1, 128.6, 127.4,
127.6, 124.3, 92.6, 89.9, 87.9, 86.8, 83.2, 72.1, 62.7, 46.0, 40.4,
34.6. FTIR (neat): 3351, 1633 cmꢀ1. HRMS (ESI) m/z: calcd for
C19H25N4O4 (M+H)+ 373.1870, found 373.1908.
4.4.1. (10S,30R,40R,50R,70R)-N3-Dimethylaminomethylidine-1-{30-
dimethoxytrithyloxymethyl-40-O-(N,N-diisopropyl-b-
cyanoethylphosphoramidyl)-10-phenyl-20,60-
dioxabicyclo[3.3.0]oct-70-yl}-3-aminopyrazole (5a)
(WNA-a
AP) 1H NMR (400 MHz, CDCl3): d 7.74 (1H, s), 7.67 (1H,
d, J = 7.3 Hz), 7.50–7.15 (12H, m), 6.78 (4H, dd, J = 1.5, 8.8 Hz), 6.60
(1H, t, J = 7.0 Hz), 5.67 (1H, s), 4.69–4.65 (1H, m), 4.50 (1H, d,
J = 4.8 Hz), 4.38–4.37 (1H, m), 3.99–3.87 (1H, m), 3.76 (6H, s),
3.77 (6H, s), 3.76–3.17 (7H, m), 3.05–3.01 (6H, m), 2.88–2.82
(1H, m), 1.06–0.78 (9H, m). 31P NMR (162 MHz, CDCl3): 149.32,
146.96. FTIR (neat): 2836, 1635, 1508, 1251 cmꢀ1. ESI-MS (m/z)
876.56 [M+H]+.
4.3.2. (10S,30R,40R,50R,70S)-N3-Dimethylaminomethylidine-1-(40-
hydroxy-30-hydroxymethyl-10-phenyl-20,60-dioxabicyclo[3.3.0]-
oct-70-yl)-3-aminopyrazole (4b)
(WNA-bAP) 1H NMR (400 MHz, CDCl3): d 7.75 (1H, s), 7.65 (2H,
d, J = 7.6 Hz), 7.46 (1H, s), 7.36 (2H, t, J = 7.6 Hz), 7.26 (1H, t,
J = 7.3 Hz), 6.66 (1H, t, J = 7.0 Hz), 5.73 (1H, s), 4.86 (1H, d,
J = 3.6 Hz), 4.07 (1H, d, J = 2.4 Hz), 4.01 (1H, d, J = 3.9 Hz), 3.99
(1H, dd, J = 3.3, 12.0 Hz), 3.80 (1H, dd, J = 4.9, 12.0 Hz), 3.45 (1H,
dd, J = 7.9, 13.9 Hz), 3.06 (6H, s), 2.82 (1H, dd, J = 6.1, 13.9 Hz).
13C NMR (125 MHz, CDCl3): 154.9, 151.9, 140.8, 140.3, 128.5,
127.6, 125.3, 92.4, 89.9, 87.9, 85.5, 84.0, 73.1, 63.2, 47.5, 40.4,
4.4.2. (10S,30R,40R,50R,70S)-N3-Dimethylaminomethylidine-1-{30-
dimethoxytrithyloxymethyl-40-O-(N,N-diisopropyl-b-
cyanoethylphosphoramidyl)-10-phenyl-20,60-
dioxabicyclo[3.3.0]oct-70-yl}-3-aminopyrazole (5b)
(WNA-bAP) 1H NMR (400 MHz, CDCl3): d 7.89 (1H, d, J = 7.0 Hz),
7.82 (1H, d, J = 7.0 Hz), 7.75 (0.5H, s), 7.68 (0.5H, s), 7.50–7.44