L. Wang et al. / Bioorganic & Medicinal Chemistry xxx (2017) xxx–xxx
7
1
8
28.22, 128.00, 127.14, 122.53, 114.99, 113.29, 106.38, 87.24,
6.59, 75.16, 73.54, 67.86, 64.11, 55.36, 40.39, 38.17. HRMS (ESI-
38.17. HRMS (ESI-TOF, m/z): Calcd. For C44
896.2151. Found: 896.2118.
General procedure of synthesis of phosphoramidite compound
(8a-c)
To a dried flask was added the corresponding compound (1.0
equivalent.) in anhydrous CH Cl under an Ar atmosphere. DIPEA
(6.0 equivalents.), and iPr NPCl(OCH CH CN) (3.0 equivalents.)
were successively added to the mixture at 0 °C. The reaction mixture
was stirred at 0 °C for 30 min, which was diluted by CH Cl , washed
with a sat. NaHCO solution and brine, dried over Na SO and puri-
fied by column chromatography (CHCl /MeOH = 100/0 to 94/6), and
precipitated from hexane at ꢂ78 °C to yield a white powder.
H
43IN
5
O
8
[M+H]+:
+
TOF, m/z): Calcd. For C44
Found: 804.2744, 806.2746.
5 8
H44ClN O [M+H] : 804.2795, 806.2794.
0
0
0
4
.1.14. 2 -Deoxy-5 -O-(4,4 -dimethoxytrityl)-1-methyl-4-N-(5-
bromo-6-phenoxyacetadepyridin-3-yl)pseudocytidine (7b)
Protection of amino group (0.24 g, 0.25 mmol, 71%). IR (cm ):
2
2
2
2
2
ꢂ1
1
2
927.8, 1662.3, 1508.2, 1250.9. H NMR (500 MHz, CDCl
3
) d
2
2
(
ppm) 8.93 (brs, 1H), 8.56 (s, 1H), 8.50 (s, 1H), 8.24 (d, J = 2.5 Hz,
3
2
4
1
6
H), 7.36–7.33 (m, 4H), 7.25–7.18 (m, 7H), 7.07–6.99 (m, 3H),
.78 (dd, J = 9.0, 2.0 Hz, 4H), 5.02 (dd, J = 10.75, 4.75 Hz, 1H), 4.73
3
(
s, 2H), 4.39 (d, J = 5.5 Hz, 1H), 4.08 (m, 1H), 3.75 (s, 3H), 3.74 (s,
0
0
0
0
3
H), 3.43 (s, 3H), 3.30–3.24 (m, 2H), 2.18–2.16 (m, 1H), 2.07 (dd,
4.1.16. 2 -Deoxy-5 -O-(4,4 -dimethoxytrityl)-3 -O-[2-cyanoethoxy
(diisopropylamino)phosphino]-1-methyl-4-N-(5-chloro-6-
phenoxyacetamidepyridin-3-yl)pseudocytidine (8a)
1
3
J = 12.25, 4.75 Hz, 1H), 0.87 (s, 9H), 0.04 (s, 3H), 0.01 (s, 3H).
NMR (125 MHz, CDCl ) d (ppm) 161.71, 158.76, 157.22, 156.22,
3
C
ꢂ1
1
1
1
4
44.40, 143.75, 143.54, 140.75, 135.54, 134.99, 132.80, 130.15,
30.13, 129.99, 128.23, 128.02, 127.19, 122.49, 115.00, 113.32,
13.31, 105.67, 88.02, 86.67, 75.95, 73.65, 67.95, 63.64, 55.39,
0.76, 38.20, 25.90, 18.11, ꢂ4.52, ꢂ4.58. HRMS (ESI-TOF, m/z):
Compound 8a (0.13 g, 0.13 mmol, 47%). IR (cm ): 2967.6,
1662.2, 1507.4, 1249.2, 913.1. H NMR (500 MHz, CDCl
1
3
) d (ppm)
8.92 (brs, 1H), 8.57 (s, 0.5 H), 8.51 (s, 0.5H), 8.46 (s, 0.5H), 8.42
(s, 0.5H), 8.22 (t, J = 2.5 Hz, 1H), 7.36–7.31 (m, 5H), 7.25–7.18 (m,
6H), 7.07–6.99 (m, 3H), 6.79–6.75 (m, 4H), 5.10–5.05 (m, 1H),
4.73 (s, 2H), 4.62–4.56 (m, 1H), 4.31–4.24 (m, 1H), 3.75 (s, 3H),
3.74 (s, 3H), 3.62–3.50 (m, 4H), 3.40 (s, 1.5H), 3.39 (s, 1.5H),
3.36–3.26 (m, 2H), 2.77–2.74 (m, 1H), 2.65–2.58 (m, 1H), 2.48–
+
Calcd. For C50
5
H57BrN O
8
Si [M+H] : 962.3154, 964.3147. Found:
9
62.3140, 964.3106.
ꢂ1
Desilylation: Compound 7b (0.20 g, 0.24 mmol, 95%). IR (cm ):
1
3
3
321.0, 1660.4, 1508.1, 1248.9, 912.0. H NMR (500 MHz, CDCl ) d
1
3
(
ppm) 8.97 (brs, 1H), 8.56 (d, J = 2.0 Hz, 1H), 8.49 (s, 1H), 8.33 (d,
J = 2.5 Hz, 1H), 7.36–7.32 (m, 4H), 7.25–7.15 (m, 7H), 7.06–6.99
m, 3H), 6.77 (d, J = 8.0 Hz, 4H), 5.11 (dd, J = 10.5, 5.0 Hz, 1H),
3
2.30 (m, 2H), 1.20–1.11 (m, 12H). C NMR (125 MHz, CDCl ) d
(ppm) 158.76, 157.30, 156.31, 144.51, 144.13, 143.83, 139.82,
135.58, 131.72, 130.19, 130.16, 130.13, 128.28, 128.21, 128.01,
127.17, 122.52, 115.00, 113.31, 86.63, 75.58, 67.96, 63.72, 58.48,
(
4
3
.68 (s, 2H), 4.49 (d, J = 5.5 Hz, 1H), 4.16–4.13 (m, 1H), 3.74 (s,
H), 3.73 (s, 3H), 3.36 (s, 3H), 3.29–3.28 (m, 2H), 2.29 (dd,
58.33, 55.38, 45.50, 45.45, 43.44, 43.35, 38.22, 38.15, 31.73,
1
3
31
J = 10.0, 5.0 Hz, 1H), 2.24–2.17 (m, 1H), 2.00 (s, 1H). C NMR
125 MHz, CDCl ) d (ppm) 161.40, 158.73, 157.15, 156.38, 144.52,
43.77, 143.27, 140.57, 135.65, 135.63, 134.15, 133.15, 130.16,
3
24.71, 22.79. P-NMR (202 MHz, CDCl ) d (ppm) 148.96, 148.13.
+
(
3
HRMS (ESI-TOF, m/z): Calcd. For
53 7 9
C H59ClN O PNa [M+Na] :
1
1
1
4
1026.3693, 1028.3699. Found: 1026.3682, 1028.3697.
30.13, 129.99, 128.22, 128.01, 127.16, 122.53, 114.99, 113.21,
06.26, 87.24, 86.60, 77.36, 75.24, 73.54, 67.86, 64.07, 55.37,
0.39, 38.18. HRMS (ESI-TOF, m/z): Calcd. For C44
0
0
0
0
4.1.17. 2 -Deoxy-5 -O-(4,4 -dimethoxytrityl)-3 -O-[2-cyanoethoxy
(diisopropylamino)phosphino]-1-methyl-4-N-(5-bromo-6-
phenoxyacetamidepyridin-3-yl)pseudocytidine (8b)
5 8
H43BrN O [M
+
+H] : 848.2290, 850.2280. Found: 848.2269, 850.2250.
ꢂ1
Compound 8b (0.20 g, 0.19 mmol, 81%). IR (cm ): 2974.9,
1662.4, 1507.4, 1248.9, 912.9. H NMR (500 MHz, CDCl
0
0
0
1
4
6
.1.15. 2 -Deoxy-5 -O-(4,4 -dimethoxytrityl)-1-methyl-4-N-(5-iodo-
-phenoxyacetadepyridin-3-yl)pseudocytidine (7c)
3
) d (ppm)
8.97 (brs, 1H), 8.67 (s, 0.5 H), 8.60 (s, 0.5H), 8.43 (s, 0.5 H), 8.39
(s, 0.5H), 8.27–8.26 (m, 1H), 7.59 (s, 0.5H), 7.37–7.31 (m, 5H),
7.25–7.18 (m, 6H), 7.07–7.00 (m, 3H), 6.79–6.75 (m, 4H), 5.10–
5.05 (m, 1H), 4.72 (s, 2H), 4.62–4.54 (m, 1H), 4.31–4.23 (m, 1H),
3.74 (s, 3H), 3.73 (s, 3H), 3.56–3.47 (m, 4H), 3.40 (s, 1.5H), 3.39
(s, 1.5H), 3.30–3.27 (m, 2H), 2.65–2.58 (m, 1H), 2.49–2.46 (m,
ꢂ1
Protection of amino group (0.32 g, 0.31 mmol, 75%). IR (cm ):
1
2
927.2, 1665.3, 1507.4, 1249.5. H NMR (500 MHz, CDCl
3
) d
(
ppm) 8.74 (brs, 1H), 8.51 (s, 1H), 8.34 (brs, 1H), 8.21 (d,
J = 2.5 Hz, 1H), 7.26–7.21 (m, 4H), 7.15–7.07 (m, 7H), 6.97–6.91
m, 3H), 6.68 (dd, J = 8.75, 1.25 Hz, 4H), 4.91 (dd, J = 11.0, 5.0 Hz,
(
1
3
1
3
H), 4.61 (s, 2H), 4.29 (d, J = 5.5 Hz, 1H), 3.97 (m, 1H), 3.65 (s,
H), 3.64 (s, 3H), 3.32 (s, 3H), 3.21 (dd, J = 10.5, 4.5 Hz, 1H), 3.16
1H), 2.31–2.04 (m, 2H), 1.20–1.10 (m, 12H). C NMR (125 MHz,
CDCl ) d (ppm) 161.41, 158.75, 157.19, 156.24, 144.45, 144.11,
3
(
7
dd, J = 10.5, 4.5 Hz, 1H), 2.13–2.07 (m, 1H), 1.97 (dd, J = 12.0,
143.80, 143.49, 140.73, 140.56, 135.56, 134.91, 130.18, 130.15,
130.12, 128.27, 128.21, 128.04, 128.01, 127.19, 122.50, 117.02,
114.99, 113.30, 86.77, 86.41, 75.51, 67.93, 63.75, 63.68, 58.33,
58.29, 55.37, 45.50, 45.45, 43.44, 43.36, 38.20, 38.13, 31.72,
1
3
.0 Hz, 1H), 0.77 (s, 9H), ꢂ0.07 (s, 3H), ꢂ0.10 (s, 3H). C NMR
(
125 MHz, CDCl
3
) d (ppm) 161.71, 158.75, 157.24, 156.23, 146.01,
1
1
8
2
44.36, 143.79, 141.97, 141.38, 135.53, 132.60, 130.14, 130.12,
29.98, 128.22, 128.02, 127.20, 115.02, 113.33, 113.31, 105.57,
8.00, 86.65, 76.02, 73.61, 67.87, 63.56, 55.40, 40.79, 38.18,
3
1
24.76, 24.70, 23.12, 23.04. P-NMR (202 MHz, CDCl
148.92, 148.14. HRMS (ESI-TOF, m/z): Calcd. For: C53 59BrN
[M+Na] : 1070.3187, 1072.3181. Found:1070.3201, 1072.3183.
3
) d (ppm)
H
7 9
O
PNa
+
5.89, 18.09, ꢂ4.53, ꢂ4.59. HRMS (ESI-TOF, m/z): Calcd. For C50
57
H -
+
IN
5
O
8
Si [M+H] : 1010.3016. Found: 1010.3017.
Desilylation: Compound 7c (0.23 g, 0.26 mmol, 81%). IR (cm ):
362.2, 2932.0, 1660.1, 1508.2, 1248.8, 912.5. 1H NMR (500 MHz,
CDCl ) d (ppm) 8.89 (brs, 1H), 8.62 (d, J = 2.0 Hz, 1H), 8.44 (s,
ꢂ1
0
0
0
0
4.1.18. 2 -Deoxy-5 -O-(4,4 -dimethoxytrityl)-3 -O-[2-cyanoethoxy
(diisopropylamino)phosphino]-1-methyl-4-N-(5-iodo-6-
phenoxyacetamidepyridin-3-yl)pseudocytidine (8c)
3
3
ꢂ1
1
7
5
1
H), 8.42 (d, J = 2.5 Hz, 1H), 7.36–7.31 (m, 4H), 7.26–7.15 (m,
H), 7.06–6.99 (m, 3H), 6.78 (d, J = 8.2 Hz, 4H), 5.10 (dd, J = 10.5,
.0 Hz, 1H), 4.66 (s, 2H), 4.49 (d, J = 5.5 Hz, 1H), 4.15–4.13 (m,
H), 3.74 (s, 6H), 3.35 (s, 3H), 3.29 (d, J = 4.5 Hz, 2H), 2.29 (dd,
Compound 8c (0.20 g, 0.18 mmol, 71%). IR (cm ): 2968.2,
1662.4, 1507.5, 1248.7, 913.0. H NMR (500 MHz, CDCl
1
3
) d (ppm)
8.87 (brs, 1H), 8.73 (s, 0.5 H), 8.66 (s, 0.5H), 8.33–8.32 (m, 1H),
7.59 (s, 1H), 7.37–7.30 (m, 5H), 7.25–7.18 (m, 6H), 7.07–7.02 (m,
3H), 6.79–6.75 (m, 4H), 5.10–5.05 (m, 1H), 4.71 (s, 2H), 4.62–4.56
(m, 1H), 4.31–4.24 (m, 1H), 3.75 (s, 3H), 3.74 (s, 3H), 3.61–3.47
(m, 4H), 3.40 (s, 1.5H), 3.39 (s, 1.5H), 3.38–3.27 (m, 2H), 2.65–
2.60 (m, 1H), 2.54–2.46 (m, 1H), 2.32–2.23 (m, 2H), 1.20–1.10
1
3
J = 12.5, 5.5 Hz, 1H), 2.21–2.15 (m, 1H), 2.00 (s, 1H). C NMR
125 MHz, CDCl ) d (ppm) 161.34, 158.72, 157.17, 156.46, 145.70,
44.54, 143.81, 141.67, 140.92, 135.68, 135.66, 133.04, 130.17,
(
3
1
1
1
30.13, 129.99, 128.24, 128.02, 127.16, 122.50, 115.02, 113.31,
06.38, 87.20, 86.56, 75.16, 73.54, 67.79, 64.07, 55.39, 40.48,
1
3
3
(m, 12H). C NMR (125 MHz, CDCl ) d (ppm) 161.45, 158.76,