N. F. Zakirova et al. / Bioorg. Med. Chem. 20 (2012) 5802–5809
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400 MHz): d 7.78 (s, 1H, H-8), 5.37 (s, 2H, H-10), 3.86–3.88 (m, 2H,
H-30), 3.71–3.73 (m, 2H, H-20), 3.43–3.45 (m, 8H, CH2OCH2), 2.77–
2.79 (m, 8H, CH2NCH2). 31P-NMR (D2O, 162 MHz): d 15.92. 13C-
NMR (D2O, 101 MHz): d 159.67 (C-4), 154.64 (C-2), 152.08 (C-6),
138.66 (C-8), 117.44 (C-5), 73.04 (C-10), 70.60 (d, J = 8.04 Hz, C-
30), 66.44 (d, J = 4.92 Hz, C-20), 64.39 (d, J = 4.01 Hz, CH2OCH2),
45.21 (d, J = 3.86 Hz, CH2NCH2). MS (ESI+) m/z 443,1711 (M).
0.66 mg). Purification and freeze-drying gave 8.8 mg (10%) of (10)
as a white lyophilisate. UV (H2O): kmax 250 nm (9700). 1H-NMR
(D2O, 400 MHz): d 7.79 (s, 1H, H-8), 5.34 (s, 2H, H-10), 3.92–3.96
(m, 2H, H-30), 3.68–3.72 (m, 2H, H-20), 3.22 (t, 2H, J = 6.08 Hz,
(CH3)2NCH2CH2NH), 2.80 (t, 2H, J = 6.08 Hz, (CH3)2NCH2CH2NH),
2.68 (s, 6H, CH3N). 31P-NMR (D2O, 162 MHz): 10.86. 13C-NMR
(D2O, 101 MHz): d 160.94 (C-4), 157.11 (C-2), 153.18 (C-6), 143.10
(C-8), 119.01 (C-5), 74.91 (C-10), 69.35 (d, J = 5.27 Hz, C-30), 65.42
(d, J = 6.99 Hz, C-20), 58.56 (d, J = 4.21 Hz, (CH3)2 NCH2CH2), 45.72
((CH3)N), 42.25 (d, J = 2.51 Hz, (CH3)2NC H2CH2). HRMS (ESIꢀ) calcd
for C12H18N6O5P (M-NH4)ꢀ 374,1342 (M-NH4)ꢀ; found 374,1322.
4.2.7. 9-(2-Hydroxyethoxymethyl)guanine, phosphoromonomo
rpholidate (7)
Product (7) was prepared according to standard procedure A
from ACV (50.0 mg, 0.22 mmol), phosphorus oxychloride (51.0
mg, 0.33 mmol) and morpholine (57.5 mg, 0.66 mmol). Purification
and freeze-drying gave 40 mg (45%) of (7) as a white lyophilisate.
UV (H2O): kmax 250 nm (10200). 1H-NMR (D2O, 400 MHz): d 7.90
(s, 1H, H-8), 5.47 (s, 2H, H-10), 3.83–3.87 (m, 2H, H-30), 3.70–3.73
(m, 2H, H-20), 3.52 (t, 4H, J = 4.40 Hz, CH2OCH2), 2.84 (dt, 4H,
J = 4.40 Hz, J = 4.80 Hz, CH2NCH2). 31P-NMR (D2O, 162 MHz): d
8.18. 13C-NMR (D2O, 101 MHz): d 160.21 (C-4), 155.07 (C-2),
152.94 (C-6), 140.28 (C-8), 118.01 (C-5), 72.81 (C-10), 70.05 (d,
J = 8.21 Hz, C-30), 65.84 (d, J = 5.08 Hz, C-20), 63.51 (d, J = 4.11 Hz,
CH2OCH2), 46.51 (d, J = 3.79 Hz, CH2NCH2). MS(ESI+) m/z 374,1119
(M+H+), 396,0944 (M+Na+), 412,0685 (M+K+); HRMS (ESIꢀ) calcd
for C12H18N6O6P (M-NH4)ꢀ:373,1026 (M-NH4)ꢀ; found 373,0085.
4.2.11. 9-(2-Hydroxyethoxymethyl)guanine, phosphoromono-
3-dimethylamino-1-propylamidate (11)
Product (11) was prepared according to standard procedure A
from ACV (50.0 mg, 0.22 mmol), phosphorus oxychloride (51.0
mg, 0.33 mmol) and 3-dimethylamino-1-propylamine (67.5 mg,
0.66 mmol). Purification and freeze-drying yielded 9.8 mg (11%) of
(11) as a white lyophilisate. UV (H2O): kmax 250 nm (9600). 1H-
NMR (D2O, 400 MHz): d 7.79 (s, 1H, H-8), 5.34 (s, 2H, H-10), 3.92–
3.96 (m, 2H, H-30), 3.68–3.72 (m, 2H, H-20), 3.65-3.69 (m, 2H,
CH2CH2CH2NH), 3.00 (t, 2H, J = 6.78 Hz, (CH3)2NCH2CH2CH2), 2.73
(s, 6H, (CH3)2N), 1.71–1.79 (m, 2H, CH2CH2CH2NH). 31P-NMR (D2O,
162 MHz): d 10.55. 13C-NMR (D2O, 101 MHz): d 162.01 (C-4),
156.83 (C-2), 154.74 (C-6), 142.83 (C-8), 120.01 (C-5), 75.02 (C-10),
71.49 (d, J = 4.76 Hz, C-30), 65.89 (d, J = 8.03 Hz, C-20), 54.61 (d,
J = 4.12 Hz, (CH3)2NCH2CH2CH2), 45.17 ((CH3)2N), 43.24 (d, J =
2.57 Hz, (CH3)2NCH2CH2CH2), 26.97 (d, J = 2.29 Hz, (CH3)2NCH2
CH2CH2). HRMS (ESI-) calcd for C13H20N6O5P (M-NH4)ꢀ 388,1499
(M-NH4)ꢀ; found 388,1465.
4.2.8. 9-(2-Hydroxyethoxymethyl)guanine, phosphoromono-N-
morpholinoethylamidate (8)
Product (8) was prepared according to standard procedure A
from ACV (50.0 mg, 0.22 mmol), phosphorus oxychloride (51.0 mg,
0.33 mmol) and 2-morpholinoethylamine (86.0 mg, 0.66 mmol).
Purification and freeze-drying gave 11.5 mg (12%) of (8) as a white
lyophilisate. UV (H2O): kmax 250 nm (9800). 1H-NMR (D2O,
400 MHz): d 7.79 (s, 1H, H-8), 5.34 (s, 2H, H-10), 3.86–3.89 (m, 2H,
H-30), 3.81 (br s, 4H, CH2OCH2), 3.73–3.75 (m, 2H, H-20), 2.92–2.98
(m, 6H, CH2CH2NH + CH2NCH2), 2.81–2.87 (m, 2H, CH2CH2NH).
31P-NMR (D2O, 162 MHz): d 9.26. 13C-NMR (D2O, 101 MHz): d
159.08 (C-4), 154.39 (C-2), 153.38 (C-6), 140.99 (C-8), 118.14 (C-
5), 74.81 (C-10), 69.34 (d, J = 5.56 Hz, C-30), 66.71 (d, J = 7.64 Hz, C-
20), 66.37 (2s, CH2OCH2), 53.61 (d, J = 2.58 Hz, CH2CH2NH), 53.07
(2s, CH2NCH2), 43.11 (d, J = 4.04 Hz, CH2CH2NH). HRMS (ESIꢀ) calcd
for C14H20N6O6P (M-NH4)ꢀ 416,1448 (M-NH4)ꢀ; found 416,1419.
4.2.12. 9-(2-Hydroxyethoxymethyl)guanine, phosphorodihexyla
midate (12)
Product (12) was prepared according to standard procedure B
from ACV (50.0 mg, 0.22 mmol), phosphorus oxychloride
(51.0 mg, 0.33 mmol) and hexylamine (223 mg, 2.2 mmol). Accord-
ing to NMR data product (12) contained ACV admixture (1) in the
quantity of 30%. UV (H2O): kmax 250 nm (9600). 1H-NMR (D2O,
400 MHz): d 7.93 (s, 1H, H-8), 5.56 (s, 2H, H-10), 3.87–3.91 (m,
2H, H-30), 3.72–3.76 (m, 2H, H-20), 2.46–2.53 (m, 4H, CH2NH),
1.07–1.24 (m, 16H, CH3(CH2)4CH2NH), 0.78 (t, 3H, J = 7.32, CH3).
31P-NMR (D2O, 162 MHz): d 16.05.
4.2.9. 9-(2-Hydroxyethoxymethyl)guanine, phosphoromono-
dipropylamidate (9)
4.2.13. 9-(2-Hydroxyethoxymethyl)guanine, phosphoromonohe
xylamidate (13)
Product (9) was prepared according to standard procedure A
from ACV (50.0 mg, 0.22 mmol), phosphorus oxychloride
(51.0 mg, 0.33 mmol) and dipropylamine (67 mg, 0.66 mmol).
Purification and freeze-drying yielded 36.5 mg (41%) of (9) as a
white lyophilisate. UV (H2O): kmax 249 nm (10000). 1H-NMR
(D2O, 400 MHz): d 7.91 (s, 1H, H-8), 5.48 (s, 2H, H-10), 3.77–3.81
(m, 2H, H-30), 3.71–3.73 (m, 2H, H-20), 2.63–2.70 (m, 4H,
CH2CH2CH3), 1.26–1.36 (m, 4H, CH2CH2CH3), 0.71 (t, 6H,
J = 7.48 Hz, CH2CH2CH3). 31P-NMR (D2O, 162 MHz): d 10.76. 13C-
NMR (D2O, 101 MHz): d 161.67 (C-4), 156.80 (C-2), 154.47 (C-6),
142.78 (C-8), 118.92 (C-5), 75.70 (C-10), 71.88 (d, J = 7.68 Hz, C-
30), 65.89 (d, J = 5.45 Hz, C-20), 50.73 (d, J = 3.43 Hz, CH2CH2CH3),
24.24 (d, J = 2.22 Hz, CH2CH2CH3), 13.45 (CH2CH2CH3). MS (ESI+)
m/z 388,1624 (M-NH3), 410,1443 (M-NH4+Na), 426,1183 (M-
NH4+K); HRMS (ESIꢀ) calcd for C14H24N6O5P (M-NH4)ꢀ: 387,1546
(M-NH4)ꢀ; found 387,1508.
Product (13) was prepared according to standard procedure A
from ACV (50.0 mg, 0.22 mmol), phosphorus oxychloride (51.0
mg, 0.33 mmol) and hexylamine (67.0 mg, 0.66 mmol). Purification
and freeze-drying yielded 15 mg (17%) of (13) as a white lyophili-
sate. UV (H2O): kmax 250 nm (9700). 1H-NMR (D2O, 400 MHz): d
7.92 (s, 1H, H-8), 5.49 (s, 2H, H-10), 3.84–3.89 (m, 2H, H-30), 3.71–
3.74 (m, 2H, H-20), 2.56–2.62 (m, 2H, CH2NH), 1.22–1.29 (m, 4H,
CH3(CH2)2CH2CH2CH2NH), 1.07–1.13 (m, 4H, CH3CH2CH2(CH2)3NH),
0.77 (t, 3H, J = 7.32 Hz, CH3). 31P-NMR (D2O, 162 MHz): d 8.56. 13C-
NMR (D2O, 101 MHz): d 164.08 (C-4), 156.37 (C-2), 155.01 (C-6),
141.14 (C-8), 119.92 (C-5), 74.78 (C-10), 69.88 (d, J = 5.11 Hz, C-30),
65.03 (d, J = 8.04 Hz, C-20), 45.17 (d, J = 3.33 Hz, CH2(CH2)4CH3),
31.75 ((CH2)3CH2CH2CH3), 28.82 (d, J = 2.2 Hz, CH2CH2(CH2)3CH3),
25.79 (d, J = 2.5 Hz, (CH2)2CH2(CH2)2CH3), 22.37 ((CH2)4CH2CH3),
14.14 (CH3). HRMS (ESIꢀ) calcd for C14H24N6O5P (M-NH4)ꢀ:
387,1546 (M-NH4)ꢀ; found 387,1578.
4.2.10. 9-(2-Hydroxyethoxymethyl)guanine, phosphoromono-
N,N-dimethylaminoethylamidate (10)
4.3. Chemical stability
Product (10) was prepared according to standard procedure A
from ACV (50.0 mg, 0.22 mmol), phosphorus oxychloride
(51.0 mg, 0.33 mmol) and dimethylaminoethylamine (58 mg,
Hydrolysis of the compounds (3-13) was performed in acidic
(pH 2, KCl and HCl, 25 ml of 0.2 M KCl and 6.5 ml of 0.2 M HCl