214
I.B. Yanachkov et al. / European Journal of Medicinal Chemistry 107 (2016) 204e218
0
0
ꢁ
(
4
4
300 MHz, D
2
O)
d
8.29e7.95 (ms, 2H, H-8), 5.89e5.82 (m, 2H, H-1 ),
(10.3%), 973.0 (8.6%), 973.9 (1.8%); calculated for [MꢁH] ,
e
.81e4.31 (1H, multiple t, CHeCl), 4.67e4.55 (m, 2H, H-2 ),
21 3 10 2
C H26Cl N O16P4S , m/z: 966.9 (89.1%), 967.9 (25.8%), 968.9
0
0
.43e4.33 (m, 2H, H-3 ), 4.24e4.16 (m, 2H, H-4 ), 4.16e4.00 (m, 4H,
(100.0%), 969.9 (28.1%), 970.9 (42.3%), 971.9 (11.4%), 972.9 (8.3%),
973.9 (2.1%); purity, 99.3%, RT RP HPLC: 7.60, 7.88, 8.12, 8.33 min
(gradient from 0 to 30% MeCN in 20 mM aqueous triethylammo-
nium acetate for 15 min); IE HPLC: 6.7, 6.9 min.
0
H-5 ), 2.98e2.81 (m, 4H, SCH
0
4
2
), 1.63e1.42 (m, 4H, SCH
2
CH
2
),
31
.87e0.78 (multiple t, 6H, CH
3
); P NMR (121 MHz, D
2
O), ppm:
1
4
2
3
e
3.8e42.5 (m, P þ P ), 3.1e1.8 (m, P þ P ); MS (ESI ), observed, m/
z: 1047.0 (100%), 1048.0 (36.7%), 1048.9 (55.3%), 1049.9, (18.4%),
ꢁ
1
C
(
050.9, (10.8%), 1051.9 (4.6%); calculated for [MꢁH] ,
1
4
0
1
4
2 3
ꢁ
4.1.11. P ,P -di-(2-bromo-5 -adenosine) P ,P -dithio-P ,P -
27
H40ClN10
O
16
P
4
S
4
, m/z: 1047.0 (100.0%), 1048.0 (37.2%), 1049.0
60.0%), 1050.0 (20.5%), 1051.0 (12.4%), 1052.0 (3.6%); purity, 98.6%,
RT RP HPLC: 9.99, 10:25 min (gradient from 0 to 50% MeCN in
chloromethylenetetraphosphate, sodium salt (11)
0
Prepared by the above procedure from 2-bromoadenosine 5 -
monothiophosphate bis-triethylammonium salt (193 mg,
2
8
0 mM aqueous triethylammonium acetate for 15 min); IE HPLC:
.4, 8.6 min.
0
.30 mmol) and disodium di-(1-imidazolyl)chloromethylene-bis-
phosphonate (35.5 mg, 0.1 mmol). Yield: 76.7 mg, 66%; H NMR
1
1
4
0
(300 MHz, D
2
O)
d
8.48e8.33 (multiple s, 2H, H-8), 6.02e5.96 (m,
4
.1.8. P ,P -di-(2-(3,3,3-trifluoropropylthylthio)-5 -adenosine)
0
0
1
4
2
3
2H, H-1 ), 5.01e4.50 (1H, multiple t, CHCl), 4.78e4.72 (m, 2H, H-2 ),
P ,P -dithio-P ,P -chloromethylenetetraphosphate, sodium salt (8)
Prepared by the above procedure from 2-(3,3,3-
trifluoropropylthio)adenosine
triethylammonium salt (208 mg, 0.30 mmol) and disodium di-(1-
imidazolyl)chloromethylene-bis-phosphonate
.1 mmol). Yield: 76 mg, 61%; H NMR (300 MHz, D
ms, 2H, H-8), 5.98e5.91 (m, 2H, H-1 ), 4.95e4.40 (1H, multiple t,
CHCl), 4.67e4.60 (m, 2H, H-2 ), 4.53e4.43 (m, 2H, H-3 ), 4.33e4.25
m, 2H, H-4 ), 4.25e4.11 (m, 4H, H-5 ), 3.24e3.12 (m, 4H, CF3CH
.70e2.50 (m, 4H, SCH ); P NMR (121 MHz, D O), ppm: 44.2e42.6
0
0
4
.63e4.57 (m, 2H, H-3 ), 4.46e4.37 (m, 2H, H-4 ), 4.37e4.22 (m, 4H,
0
31
1
4
0
H-5 ); P NMR (121 MHz, D
2
O), ppm: 44.05e42.58 (m, P þ P ),
.92e2.02 (m, P þ P ); MS (ESIe), observed, m/z: 1055.1 (42.0%),
056.2 (10.0%), 1057.0 (100.0%), 1058.0 (29.6%), 1058.9 (77.3%),
5 -monothiophosphate
bis-
2
3
2
1
(35.5
O)
mg,
1
1059.9 (20.6%), 1060.8 (24.4%), 1061.8 (6.8%), 1062.8 (2.0%); calcu-
0
(
2
d
8.28e8.10
ꢁ
ꢁ
2
, m/z: 1054.8 (41.2%),
0
lated for [MꢁH] , C21
2
H26Br ClN10
O
16
P
4
S
0
0
1055.8 (11.9%), 1056.8 (100.0%), 1057.8 (28.6%), 1058.8 (80.3%),
1059.8 (22.4%), 1060.8 (24.1%), 1061.8 (6.3%), 1062.8 (2.8%); purity,
97.8%, RT RP HPLC: 7.05, 7.29 min (gradient from 0 to 50% MeCN in
20 mM aqueous triethylammonium acetate for 15 min); IE HPLC:
0
0
(
2
(
(
1
1
(
(
2
),
3
1
2
2
1
4
2
3
e
m, P þ P ), 3.0e2.1 (m, P þ P ); MS (ESI ), observed, m/z: 1155.1
6.8, 7.1 min.
100%), 1156.1 (35.2%), 1157.0 (54.9%), 1158.0 (18.4%), 1159.0 (10.2%),
ꢁ
ꢁ
159.9 (3.2%); calculated for [MꢁH] , C27
6 10 16 4
H34ClF N O P S , m/z:
4
155.0 (100.0%),1156.0 (37.1%),1157.0 (60.0%),1158.0 (20.4%),1159.0
1
4
0
1
4
2 3
4
.1.12. P ,P -di-(2-iodo-5 -adenosine) P ,P -dithio-P ,P -
11.1%), 1160.0 (3.3%); purity, 96.1%, RT RP HPLC: 11.59, 11.87 min
gradient from 0 to 50% MeCN in 20 mM triethylammonium acetate
for 15 min); IE HPLC: 8.2, 8.5 min.
chloromethylenetetraphosphate, sodium salt (12)
Prepared by the above procedure from 2-iodoadenosine 5 -
monothiophosphate bis-triethylammonium salt (208 mg,
0
0
.30 mmol) and disodium di-(1-imidazolyl)chloromethylene-bis-
1
4
0
1
4
2 3
4
.1.9. P ,P -di-(2-pentylthio-5 -adenosine) P ,P -dithio-P ,P -
1
phosphonate (35.5 mg, 0.1 mmol). Yield: 90.6 mg, 73%; H NMR
chloromethylenetetraphosphate, sodium salt (9)
Prepared by the above procedure from 2-pentylthioadenosine
-monothiophosphate bis-triethylammonium salt (200 mg,
.30 mmol) and disodium di-(1-imidazolyl)chloromethylene-bis-
phosphonate (35.5 mg, 0.1 mmol). Yield: 75 mg, 64%; H NMR
(300 MHz, D
2
O)
d
8.38e8.16 (multiple s, 2H, H-8), 5.89e5.84 (m,
0
2
2
4
3
H, H-1 ), 4.93e4.39 (1H, multiple t, CHeCl), 4.64e4.57 (m, 2H, H-
), 4.52e4.45 (m, 2H, H-3 ), 4.34e4.27 (m, 2H, H-4 ), 4.27e4.14 (m,
0
5
0
0
0
0
0
31
1
4
H, H-5 ); P NMR (121 MHz, D
.13e2.02 (m, P þ P ); MS (ESI ), observed, m/z: 1151.0 (100.0%),
2
O), ppm: 43.95e42.45 (m, P þ P ),
1
2
3
e
(
300 MHz, D
2
O)
d
8.26e8.12 (multiple s, 2H, H-8), 5.98e5.93 (m,
1152.0 (29.7%), 1152.9 (46.2%), 1153.9 (13.7%), 1154.9 (5.8%), 1155.9
0
2
2
4
1
(
H, H-1 ), 4.89e4.39 (1H, multiple t, CHeCl), 4.73e4.64 (m, 2H, H-
ꢁ
e
(
1.3, %); calculated for [MꢁH] , C21
100.0%),1151.8 (28.9%),1152.8 (48.3%),1153.8 (13.1%),1154.8 (6.3%),
155.8 (1.4%); purity, 96.4%, RT RP HPLC: 9.32, 9.75, 9.95, 10.12 min
gradient from 0 to 30% MeCN in 20 mM aqueous triethylammo-
nium acetate for 15 min); IE HPLC: 6.8, 7.1 min.
2 10 16 4 2
H26ClI N O P S , m/z: 1150.8
0
0
0
), 4.52e4.45 (m, 2H, H-3 ), 4.32e4.23 (m, 2H, H-4 ), 4.23e4.11 (m,
(
1
(
0
H, H-5 ), 3.03e2.92 (m, 4H, SCH
2
), 1.63e1.50 (m, 4H, SCH
2
CH
2
),
); P NMR
O), ppm: 44.05e42.83 (m, P þ P ), 2.96e2.13 (m,
3
1
.34e1.12 (m, 8H, CH
121 MHz, D
2 2 3 3
CH CH ), 0.82e0.72 (m, 6H, CH
1
4
2
2
3
e
P þ P ); MS (ESI ), observed, m/z: 1103.3 (100%), 1104.3 (43.1%),
1
105.1 (61.8%), 1106.1 (21.8%), 1107.0 (11.0%), 1108.0 (3.2%); calcu-
ꢁ
ꢁ
1
4
6
0
1
4
2 3
lated for [MꢁH] , C31
41.6%), 1105.1 (61.8%), 1106.1 (23.1%), 1107.1 (13.3%), 1108.1 (4.1%);
purity, 95.2%, RT RP HPLC: 12.08, 12.50, 12.67 min (gradient from
to 50% MeCN in 20 mM aqueous triethylammonium acetate for
H
48ClN10
O
16
P
4
S
4
, m/z: 1103.1 (100.0%), 1104.1
4.1.13. P ,P -di-(N -methyl-5 -adenosine) P ,P -dithio-P ,P -
chloromethylenetetraphosphate, sodium salt (13)
Prepared by the above procedure from N -methyladenosine 5 -
monothiophosphate bis-triethylammonium salt (174 mg,
0.30 mmol) and disodium di-(1-imidazolyl)chloromethylene-bis-
phosphonate (35.5 mg, 0.1 mmol). Yield: 67.1 mg, 66%; H NMR
(300 MHz, D
5.98e5.91 (2H, m, H-1 ), 5.03e4.42 (1H, mt, CHeCl), 4.77e4.68 (2H,
m, H-2 ), 4.55e4.47 (2H, m, H-3 ), 4.37e4.29 (2H, m, H-4 ),
(
6
0
0
1
5 min); IE HPLC: 8.4, 8.6 min.
1
1
4
0
1
4
2 3
4
.1.10. P ,P -di-(2-chloro-5 -adenosine) P ,P -dithio-P ,P -
chloromethylenetetraphosphate, sodium salt (10)
Prepared by the above procedure from 2-chloroadenosine 5 -
monothiophosphate bis-triethylammonium salt (180 mg,
.30 mmol) and disodium di-(1-imidazolyl)chloromethylene-bis-
2
O)
d
8.38e8.16 (2H, ms, H-8), 7.98e7.92 (ms, 2H, H-2),
0
0
0
0
0
0
31
3
4.29e4.08 (4H, m, H-5 ), 2.90e2.82 (6H, ms, NCH ); P NMR
1
4
0
(121 MHz, D
2
O), ppm: 43.97e42.43 (m, P þ P ), 3.03e1.92 (m,
1
2
3
e
phosphonate (35.5 mg, 0.1 mmol). Yield: 71.5 mg, 69%; H NMR
P þ P ); MS (ESI ), observed, m/z: 927.1 (100.0%), 928.1 (31.9%),
ꢁ
(
300 MHz, D
2
O)
d
8.39e8.26 (multiple s, 2H, H-8), 5.91e5.86 (m,
929.1 (47.0%), 930.1 (19.3%), 931.0 (8.1%); calculated for [MꢁH] ,
0
e
2
2
2
4
2
9
H, H-1 ), 4.90e4.39 (1H, multiple t, CHeCl), 4.70e4.61 (m, 2H, H-
C
23
H32ClN10
O
16
P
4
S
, m/z: 927.0 (100.0%), 929.0 (49.0%), 928.0
0
0
0
), 4.53e4.46 (m, 2H, H-3 ), 4.36e4.27 (m, 2H, H-4 ), 4.27e4.11 (m,
(31.1%), 930.0 (14.2%), 931.0 (6.6%); purity, 99.2%, RT RP HPLC: 11.13,
11.63, 11.84, 12.28 min (gradient from 0 to 15% MeCN in 20 mM
aqueous triethylammonium acetate for 15 min); IE HPLC: 6.6,
6.8 min.
0
31
1
4
H, H-5 ); P NMR (121 MHz, D
2
O), ppm: 43.75e42.42 (m, P þ P ),
2
3
e
.63e1.95 (m, P þ P ); MS (ESI ), observed, m/z: 967.0 (92.2%),
68.0 (27.0%), 969.0 (100.0%), 970.0 (28.9%), 971.0 (42.0%), 972.0