(m, 5H), 5.66 (d, 1H, J ) 8.1), 6.06 (br s, 1H), 6.37 (d, 1H, J )
8.7), 6.76 (d, 1H, J ) 8.7), 7.31-7.35 (m, 10H), 7.46-7.60 (m,
3H), 8.03 (s, 1H), 8.15-8.19 (m, 2H), 8.54 (s, 1H), 9.27 (br s,
1H) ppm. 13C NMR (125 MHz, CDCl3) δ 54.9, 60.2, 62.45, 69.5
(d, J C-P ) 6), 72.1, 76.3, 76.5, 86.8, 88.5, 113.0, 124.3, 127.8,
127.9, 128.0, 128.5, 128.6, 128.7, 129.4, 132.7, 133.5, 135.6, 142.7,
149.8, 150.1, 151.4, 159.0, 164.6 ppm. 31P NMR (164 MHz,
CDCl3) δ -4.0 ppm. HRMS calcd for C39H39O9N5P [M + H]+
752.2485, found 752.2490.
2H), 4.31-4.33 (m, 2H), 4.55-4.58 (d, 1H, J ) 11.4), 4.87-4.90
(m, 1H), 5.06-5.14 (m, 4H), 5.30 (dd, 1H, J ) 4.9, 7.2), 6.13 (d,
1H, J ) 7.6), 6.54 (d, 2H, J ) 8.6), 6.93 (d, 2H, J ) 8.6), 7.31-
7.38 (m, 10H), 8.15 (s, 1H), 8.42 (s, 1H) ppm. 13C NMR (125 MHz,
CDCl3) δ 55.5, 65.5, 71.0 (d, J C-P ) 5), 71.1 (d, J C-P ) 5), 73.7,
78.3, 81.7, 85.1, 86.7, 114.4, 120.0, 129.2, 129.6, 129.7, 130.4,
130.5, 137.0, 140.9, 150.6, 153.6, 157.0, 160.6 ppm. 31P NMR
(164 MHz, CDCl3) δ -1.2, 2.0 ppm. MS calcd for C32H36O11N5P2
[M + H]+ 728.2, found (ES (+)) m/z 728.3.
N6-Ben zoyl-5′-O-[[bis(allyloxy)ph osph in yl]oxy]-3′-O-[[bis-
(b en zyloxy)p h osp h in yl]oxy]-2′-O-(4-m et h oxyb en zyl)a d e-
n osin e (7). A suspension of 6 (1.7 g, 2.3 mmol) and tetrazole
(640 mg, 9.1 mmol) in CH2Cl2 (30 mL) was chilled to 0 °C and
diallyl diisopropylphosphoramidite (2.2 mL, 9.1 mmol) was
added dropwise via syringe. The resulting cloudy solution was
stirred for 2 h before the addition of mCPBA (1.8 g, 8.3 mmol)
in portions. After an additional 1 h of stirring on ice, the reaction
was poured into sat. NaHCO3 (100 mL) and extracted with CH2-
Cl2. The combined organic layers were washed with sat. NaHCO3
and brine, dried, and concentrated to an oil that was purified
by silica gel chromatography (9:1 EtOAc:Hex) to provide a clear
glass (1.8 g, 84%). 1H NMR (300 MHz, CDCl3) δ 3.67 (s, 3H),
4.13-4.19 (m, 1H), 4.23-4.28 (m, 1H), 4.32-4.34 (m, 2H), 4.47-
4.53 (m, 4H), 4.58 (d, 1H, J ) 8.7), 4.76 (app t, 1H, J ) 7.2),
5.03-5.13 (m, 5H), 5.22 (app dd, 2H, J ) 7.0, 11.5), 5.29-5.31
(m, 1H), 5.33-5.35 (m, 1H), 5.83-5.92 (m, 2H), 6.03 (d, 1H, J )
8.0), 6.58 (d, 2H, J ) 11.0), 6.93 (d, 2H, J ) 11.0), 7.32-7.35
(m, 10H), 7.50 (app t, 2H, J ) 9.5), 7.56 (app t, 1H, J ) 9.0),
8.03 (s, 1H), 8.04 (d, 2H, J ) 9.0), 8.70 (s, 1H) ppm. 13C NMR
(125 MHz, CDCl3) δ 55.3, 65.9, 68.6, 69.9, 72.6, 74.6, 77.8, 82.3,
86.7, 113.7, 118.9, 123.2, 128.1, 128.1, 128.4, 128.7, 128.8, 128.8,
128.9, 129.0, 129.7, 132.2, 132.2, 133.0, 133.6, 135.7, 141.8, 149.5,
151.6, 152.2, 159.5, 164.8 ppm. 31P NMR (164 MHz, CDCl3) δ
-4.2, -3.6 ppm. HRMS calcd for C45H48O12N5P2 [M + H]+
912.2774, found 912.2757.
3′-[[Bis(ben zyloxy)p h osp h in yl]oxy]a d en osin e 5′-Dip h o-
p h o-(3-estr on e) (9). To a solution of 2 (50 mg, 0.14 mmol) in
DMF (400 µL) was added triethylamine (22 µL, 0.15 mmol)
followed by carbonyldiimidazole (74 mg, 0.45 mmol). The result-
ing suspension was stirred at room temperature for 3 h and
several drops of MeOH were added. After 5 min, the reaction
mixture was concentrated and coevaporated with DMF. Com-
pound 3 (30 mg, 0.09 mmol) was added along with DMF (400
µL) and the suspension was stirred at room temperature for 2
d. The reaction mixture was then diluted into 25 mM NH4OAc
(10 mL), washed with CH2Cl2 (2 mL), and purified by reversed-
phase HPLC (10 to 100% CH3CN in 25 mM NH4OAc) to provide
33 mg (43%) of 9 as an amorphous solid. 1H NMR (500 MHz,
CD3OD) δ 0.79 (s, 3H), 1.22-1.39 (m, 6H), 1.51-1.58 (m, 1H),
1.58-1.85 (m, 2H), 1.94-2.20 (m, 4H), 2.37-2.47 (m, 1H), 2.66-
2.71 (m, 2H), 3.62 (s, 3H), 4.20-4.32 (m, 4H), 4.50 (d, 1H, J )
11.4), 5.06-5.13 (m, 4H), 5.35 (dd, 1H, J ) 4.8, 7.1), 6.08 (d,
1H, J ) 7.5), 6.50 (d, 2H, J ) 8.7), 6.89 (d, 2H, J ) 8.7), 6.92-
7.06 (m, 3H), 7.32-7.48 (m, 10H), 8.12 (s, 1H), 8.60 (s, 1H) ppm.
13C NMR (125 MHz, CD3OD) δ 12.9, 21.0, 25.4, 26.0, 28.8, 31.2,
35.2, 38.0, 43.7, 48.4, 50.1, 54.2, 64.9, 69.6, 72.3, 77.0, 80.4, 83.8,
85.4, 113.0, 117.7, 118.4, 118.9, 120.3, 125.6, 127.8, 128.2, 128.3,
129.2, 134.0, 134.7, 135.7, 137.3, 140.8, 148.8, 150.5, 152.9, 159.2,
222.1 ppm. 31P NMR (164 MHz, CD3OD) δ -14.3 (d, J P-P ) 21),
-10.2 (d, J P-P ) 21), -1.2 ppm. HRMS calcd for C50H55N5O15P3
[M]- 1058.2913, found 1058.2904.
5′-O-[[Bis(allyloxy)ph osph in yl]oxy]-3′-O-[[bis(ben zyloxy)-
p h osp h in yl]oxy]-2′-O-(4-m eth oxyben zyl)a d en osin e (8). To
a solution of 7 (1.0 g, 1.1 mmol) in dry THF (8 mL) was added
n-PrNH2 (1.2 mL, 11 mmol) slowly via syringe. The solution was
stirred for 3 h and concentrated, and the resulting oil was
purified by silica gel chromatography (5% MeOH in CH2Cl2) to
provide 630 mg (68%) of an amorphous glass. 1H NMR (500 MHz,
CDCl3) δ 3.68 (s, 3H), 4.14-4.18 (m, 1H), 4.24-4.35 (m, 2H),
4.39 (d, 1H, J ) 11.6), 4.46-4.51 (m, 4H), 4.59 (d, 1H, J ) 11.6),
4.76 (app t, 1H, J ) 5.5), 5.03-5.08 (m, 4H), 5.12-5.13 (m, 1H),
5.19 (app t, 2H, J ) 10.2), 5.30 (ddd, 2H, J ) 1.4, 8.4, 9.8), 5.82-
5.89 (m, 2H), 6.01 (d, 1H, J ) 6.2), 6.61-6.63 (m, 4H), 6.98 (d,
2H, J ) 8.6), 7.28-7.34 (m, 10H), 7.80 (s, 1H), 8.26 (s, 1H) ppm.
13C NMR (125 MHz, CDCl3) δ 55.1, 65.8 (d, J C-P ) 5), 68.4 (d,
3′-P h osp h oa d en osin e 5′-Dip h osp h o-(3-estr on e) (1). Com-
pound 9 was dissolved in H2O (5 mL), 10% Pd/C (25 mg) was
added, and the suspension was shaken under 3 atm of H2 for 6
h. The mixture was then filtered through Celite, concentrated,
and purified by DEAE-sepharose anion exchange chromatogra-
phy (0 to 1 M NH4HCO3 over 400 mL) followed by reversed-
phase HPLC (0 to 30% CH3CN in NH4OAc). Lyophilization
provided 12 mg (52%) of a white solid. 1H NMR (500 MHz, D2O)
δ 0.74 (s, 3H), 1.07-1.12 (m, 3H), 1.27-1.32 (m, 2H), 1.49-1.52
(m, 1H), 1.68-1.74 (m, 3H), 1.94-2.03 (m, 8 H), 2.13-2.17 (m,
1H), 2.47-2.52 (m, 2H), 2.59-2.61 (m, 1H), 4.08-4.14 (m, 2H),
4.51 (app s, 1H), 4.72-4.47 (m, 2H), 4.85-4.89 (m, 2H), 6.04 (d,
1H, J ) 6.5), 6.70 (app s, 1H), 6.76-6.83 (m, 2H), 8.15 (s, 1H),
8.25 (s, 1H) ppm. 13C NMR (125 MHz, D2O) δ 13.2, 20.9, 21.5,
24.8, 25.6, 28.4, 30.7, 35.9, 37.3, 42.9, 48.4, 49.6, 65.1, 73.8, 74.3,
83.3, 86.2, 117.2, 118.1, 120.0, 125.7, 135.2, 137.8, 139.9, 148.8,
149.3, 151.1, 154.1, 229.1 ppm. 31P NMR (164 MHz, D2O) δ -16.4
(d, J P-P ) 16), -11.8 (d, J P-P ) 16), -0.4 ppm. HRMS calcd for
C28H35N5O14P3 [M]- 758.1399, found 758.1384.
J C-P ) 5), 68.4 (d, J C-P ) 5), 69.7 (d, J C-P ) 9), 69.7 (d, J C-P
9), 72.4, 74.5 (d, J C-P ) 5), 77.7, 81.8, 86.5, 113.6, 118.6, 120.0,
127.9, 128.0, 128.5, 128.6, 128.6, 128.7, 129.6, 132.1 (d, J C-P
)
)
6), 132.1 (d, J C-P ) 6), 135.5 (d, J C-P ) 4), 135.5 (d, J C-P ) 4),
139.1, 149.5, 152.7, 155.6, 159.4 ppm. 31P NMR (164 MHz,
CDCl3) δ -4.3, -3.8 ppm. HRMS calcd for C38H44O11N5P2 [M +
H]+ 808.2512, found 808.2516.
Ack n ow led gm en t. The Center for New Directions
in Organic Synthesis is supported by Bristol-Myers
Squibb as Sponsoring Member and Novartis as Sup-
porting Member. This research was supported by grants
from Dupont and the National Institutes of Health
(GM59907). D.E.V. was supported by NIH Biotechnol-
ogy Training Grant No. GM08352.
3′-O-[[Bis(ben zyloxy)p h osp h in yl]oxy]-2′-O-(4-m eth oxy-
ben zyl)-5′-O-p h osp h a te Ad en osin e (3). A solution of 8 (230
mg, 0.29 mmol) in dry THF (2.5 mL) was treated with nBuNH2
(170 µL, 1.7 mmol), formic acid (260 µL, 6.9 mmol), PPh3 (45
mg, 0.17 mmol), and Pd(PPh3)4 (31 mg, 0.027 mmol). The orange
solution was heated to 40 °C for 4 h, then concentrated to an oil
that was purified by silica gel chromatography (10:2:1 EtOAc:
1
MeOH:H2O) to provide 140 mg (65%) of a clear glass. H NMR
Su p p or t in g In for m a t ion Ava ila b le: 1H and 13C NMR
spectra for all obtained compounds, procedures for the syn-
thesis of compound 2 and for kinetic assays, and general
experimental details. This material is available free of charge
(500 MHz, CDCl3) δ 3.35 (s, 6H), 3.66 (s, 3H), 4.06-4.07 (m,
(26) Ehrhardt, D. W.; Atkinson, E. M.; Faull, K. F.; Freedberg, D.
I.; Sutherlin, D. P.; Armstrong, R.; Long, S. R. J . Bacteriol. 1995, 177,
6237.
(27) Anderson, C.; Freeman, J .; Luca, L. H.; Farley, M.; Dalhoumi,
H.; Widlanski, T. S. Biochemistry 1997, 36, 2586.
J O0260443
J . Org. Chem, Vol. 68, No. 1, 2003 173