Journal of Medicinal Chemistry
Article
D2O) δ 15.6, 0.31. HRMS (ESI) calcd for C15H24N3O13P2 [M − H]−,
516.0784; found, 516.0779.
1H), 2.08−2.02 (m, 1H), 1.92 (s, 3H), 1.92−1.89 (m, 1H), 1.58 (dt, J
= 13.2, 9.6 Hz, 1H). 13C NMR (151 MHz, D2O) δ 174.1, 165.7, 156.9,
142.6, 138.2, 129.4, 129.1, 128.9, 97.0, 90.3, 84.5, 83.4 (d, J = 9.1 Hz),
76.4 (dd, J = 155.5, 7.6 Hz), 75.0, 71.8, 69.8, 65.2 (d, J = 4.5 Hz), 56.4,
37.5, 34.0 (d, J = 10.6 Hz), 32.3 (d, J = 4.5 Hz), 22.7. 31P NMR (162
MHz, D2O) δ 15.52, 0.48. HRMS (ESI) calcd for C24H33N4O13P2 [M
− H]−, 647.1519; found, 647.1525.
Trisodium Cytidin-5′-yl-[(1S,3R,4R)-3-hydroxy-4-acetamido-1-cy-
clopentyl-phosphonato-methyl]-phosphate (4S). This compound
was prepared in the same manner as described in the preparation of 2.
Quantities: 47c (53.3 mg, 0.04 mmol), Pd/C (53.3 mg, 10%), 1,4-
cyclohexadiene (198.2 mg, 234 μL, 2.40 mmol, 97%), DMF (1.0 mL),
and eluent (H2O), affording crude product 4S (17.7 mg, 79% yield).
The diastereoisomers were further separated by preparative RP-HPLC,
converted to their sodium salt form by ion-exchange (IR 120 Na+) and
lyophilized from water to give 4Ss and 4Sl as white powder. 4Ss: RP-
HPLC, column A, 0−5 min linear gradient 0−1.0% CH3CN, H2O
(0.1% TFA), 15 mL/min flow, tR = 4.28 min; [α]3D0 = +4.0 (c = 1.9,
H2O). 1H NMR (600 MHz, D2O) δ 8.07 (d, J = 7.8 Hz, 1H), 6.20 (d,
J = 7.8 Hz, 1H), 5.94 (d, J = 3.0 Hz, 1H), 4.32−4.18 (m, 6H), 4.05−
4.00 (m, 1H), 3.97−3.94 (m, 1H), 2.69−2.67 (m, 1H), 2.24 (dt, J =
13.8, 7.2 Hz, 1H), 2.06 (dt, J = 13.8, 7.8 Hz, 1H), 1.96 (s, 3H), 1.80
(ddd, J = 13.8, 9.0, 4.8 Hz, 1H), 1.61 (dt, J = 13.2, 9.6 Hz, 1H). 13C
NMR (151 MHz, D2O) δ 174.6, 163.6, 154.0, 143.5, 96.6, 90.3, 83.7
(d, J = 9.1 Hz), 77.0, 76.6 (dd, J = 158.6, 9.1 Hz), 75.0, 69.8, 65.1 (d, J
= 4.5 Hz), 58.2, 37.0, 35.7 (d, J = 9.1 Hz), 31.9 (d, J = 4.5 Hz), 22.7
ppm. 31P NMR (162 MHz, D2O) δ 15.66, 0.56. HRMS (ESI) calcd for
C17H27N4O13P2 [M − H]−, 557.1050; found, 557.1055. 4Sl: RP-
HPLC, column A, 0−5 min linear gradient 0−1.0% CH3CN, H2O
(0.1% TFA), 15 mL/min flow, tR = 4.80 min; [α]3D0 = −3.9 (c = 1.0,
H2O). 1H NMR (600 MHz, D2O) δ 7.97 (d, J = 7.8 Hz, 1H), 6.12 (d,
J = 7.2 Hz, 1H), 5.99 (d, J = 4.2 Hz, 1H), 4.36 (t, J = 5.4 Hz, 1H), 4.30
(t, J = 4.2 Hz, 1H), 4.27−4.23 (m, 3H), 4.19 (ddd, J = 11.4, 4.2, 2.4
Hz, 1H), 4.03 (dt, J = 7.8, 5.4 Hz, 1H), 3.92 (dd, J = 15.0, 7.2 Hz, 1H),
2.71−2.65 (m, 1H), 2.20 (dt, J = 13.2, 7.8 Hz, 1H), 2.16 (dt, J = 14.4,
8.4 Hz, 1H), 1.93 (s, 3H), 1.75 (ddd, J = 13.8, 9.0, 4.8 Hz, 1H), 1.54
(dt, J = 12.6, 9.6 Hz, 1H). 13C NMR (151 MHz, D2O) δ 174.6, 166.2,
157.6, 142.4, 97.2, 89.9, 83.5 (d, J = 9.1 Hz), 77.1, 76.4 (dd, J = 158.6,
9.1 Hz), 74.9, 69.9, 64.9 (d, J = 6.0 Hz), 58.1, 36.9, 33.8 (d, J = 10.6
Hz), 33.3 (d, J = 3.0 Hz), 22.6. 31P NMR (162 MHz, D2O) δ 15.65,
0.20. HRMS (ESI) calcd for C17H27N4O13P2 [M − H]−, 557.1050;
found, 557.1069.
Trisodium Cytidin-5′-yl-[(1S,3R,4R)-3-benzyloxy-4-acetamido-1-
cyclopentyl-phosphonatomethyl]-phosphate (5S). To a solution of
47c (117.7 mg, 0.089 mmol) in EtOH (2.7 mL) were added Pd/C
(235.4 mg, 10%) and 1,4-cyclohexadiene (847.0 mg, 1.0 mL, 10.68
mmol, 97%) under argon atmosphere. The reaction mixture was
stirred at room temperature until TLC showed complete consumption
of the starting material. Then the mixture was filtered and
concentrated. The residue was purified by column chromatography
on RP-18 silica gel (H2O → MeOH/H2O = 1:1) to give crude product
5S (25.2 mg, 44% yield). The diastereoisomers were further separated
by preparative RP-HPLC, converted to the form of sodium salt by ion-
exchange (IR 120 Na+) and lyophilized from water to give 5Ss and 5Sl
as white powder. 5Ss: RP-HPLC, column B, 0−18 min linear gradient
7.0−8.8% CH3CN, 100 mM NH4HCO3 buffer (pH = 8.4), 15 mL/
min flow; tR = 15.72 min; [α]3D0 = −1.7 (c = 0.7, H2O). 1H NMR (600
MHz, D2O) δ 7.94 (d, J = 7.2 Hz, 1H), 7.42−7.35 (m, 5H), 6.08 (d, J
= 7.8 Hz, 1H), 5.97 (d, J = 4.2 Hz, 1H), 4.59 (d, J = 11.4 Hz, 1H), 4.51
(d, J = 12.0 Hz, 1H), 4.32 (t, J = 5.4 Hz, 1H), 4.27−4.07 (m, 6H), 3.87
(dt, J = 7.2, 4.8 Hz, 1H), 2.68−2.60 (m, 1H), 2.20 (dt, J = 12.6, 7.8 Hz,
1H), 2.12 (dt, J = 13.8, 9.0 Hz, 1H), 1.94−1.90 (m, 1H), 1.89 (s, 3H),
1.50 (dt, J = 12.6, 10.2 Hz, 1H). 13C NMR (151 MHz, D2O) δ 174.0,
166.5, 157.9, 142.2, 138.1, 129.4, 129.1, 128.9, 97.1, 89.9, 84.2, 83.3 (d,
J = 9.1 Hz), 76.7 (dd, J = 157.0, 7.6 Hz), 75.0, 71.8, 69.7, 64.7 (d, J =
4.5 Hz), 56.4, 37.6, 34.3 (d, J = 9.1 Hz), 32.3 (d, J = 4.5 Hz), 22.6
ppm. 31P NMR (162 MHz, D2O) δ 15.55, 0.11. HRMS (ESI) calcd for
C24H33N4O13P2 [M − H]−, 647.1519; found, 647.1513. 5Sl: RP-
HPLC, column B, 0−18 min linear gradient 7.0−8.8% CH3CN, 100
mM NH4HCO3 buffer (pH = 8.4), 15 mL/min flow; tR = 16.70 min;
[α]3D0 = +0.5 (c = 0.4, H2O). 1H NMR (600 MHz, D2O) δ 7.97 (d, J =
7.8 Hz, 1H), 7.42−7.36 (m, 5H), 6.11 (d, J = 7.2 Hz, 1H), 5.94 (d, J =
3.6 Hz, 1H), 4.61 (d, J = 12.0 Hz, 1H), 4.55 (d, J = 12.0 Hz, 1H),
4.29−4.24 (m, 5H), 4.19−4.16 (m, 1H), 4.14−4.10 (m, 1H), 3.91 (dt,
J = 6.6, 4.8 Hz, 1H), 2.70−2.62 (m, 1H), 2.25 (dt, J = 13.8, 7.2 Hz,
Trisodium Cytidin-5′-yl-[(1R,3R,4R)-3-benzyloxy-4-acetamido-1-
cyclopentyl-phosphonatomethyl]-phosphate (5R). This compound
was prepared in the same manner as described in the preparation of
5S. Quantities: 47d (210.7 mg, 0.16 mmol), Pd/C (421.4 mg, 10%),
1,4-cyclohexadiene (1.61 g, 1.9 mL, 19.08 mmol, 97%), EtOH (4.8
mL), and eluent (H2O → MeOH/H2O = 1:1), affording crude
product 5R (51.8 mg, 50% yield). The diastereoisomers were further
separated by preparative RP-HPLC, converted to the form of sodium
salt by ion-exchange (IR 120 Na+), and lyophilized from water to give
5Rs and 5Rl as white powder. 5Rs: RP-HPLC, column B, 0−16 min
linear gradient 8−9.5% CH3CN, 100 mM NH4HCO3 buffer (pH =
8.4), 15 mL/min flow; tR = 11.95 min; [α]3D0 = +0.7 (c = 0.3, H2O). 1H
NMR (600 MHz, D2O) δ 7.96 (d, J = 7.8 Hz, 1H), 7.41−7.35 (m,
5H), 6.09 (d, J = 7.8 Hz, 1H), 5.93 (d, J = 3.6 Hz, 1H), 4.61 (d, J =
12.0 Hz, 1H), 4.55 (d, J = 11.4 Hz, 1H), 4.31 (t, J = 5.4 Hz, 1H),
4.28−4.17 (m, 5H), 4.10−4.07 (m, 1H), 3.91 (dt, J = 7.2, 6.6 Hz, 1H),
2.52−2.48 (m, 1H), 2.38 (dt, J = 12.6, 6.6 Hz, 1H), 2.12 (dt, J = 13.8,
9.6 Hz, 1H), 1.92 (s, 3H), 1.68−1.62 (m, 2H). 13C NMR (151 MHz,
D2O) δ 174.0, 166.9, 158.3, 142.1, 138.2, 129.4, 129.2, 128.9, 97.1,
90.4, 85.3, 83.3 (d, J = 9.1 Hz), 76.4 (dd, J = 152.5, 15.1 Hz), 75.2,
72.1, 69.4, 64.8, 55.4, 37.6, 34.2 (d, J = 7.6 Hz), 33.8 (d, J = 4.5 Hz),
22.7. 31P NMR (162 MHz, D2O) δ 14.68, 0.56. HRMS (ESI) calcd for
C24H33N4O13P2 [M − H]−, 647.1519; found, 647.1537. 5Rl: RP-
HPLC, column B, 0−16 min linear gradient 8−9.5% CH3CN, 100
mM NH4HCO3 buffer (pH = 8.4), 15 mL/min flow; tR = 12.58 min;
[α]3D0 = −3.7 (c = 0.4, H2O). 1H NMR (600 MHz, D2O) δ 7.98 (d, J =
7.8 Hz, 1H), 7.41−7.34 (m, 5H), 6.09 (d, J = 7.8 Hz, 1H), 5.90 (d, J =
3.0 Hz, 1H), 4.60 (d, J = 12.0 Hz, 1H), 4.53 (d, J = 11.4 Hz, 1H), 4.32
(t, J = 5.4 Hz, 1H), 4.28−4.19 (m, 5H), 4.05 (dt, J = 9.6, 4.8 Hz, 1H),
3.91 (dt, J = 8.4, 6.6 Hz, 1H), 2.50 (brs, 1H), 2.27 (dt, J = 12.6, 6.0 Hz,
1H), 2.17 (dt, J = 13.8, 10.2 Hz, 1H), 1.89 (s, 3H), 1.68−1.62 (m,
2H). 13C NMR (151 MHz, D2O) δ 173.9, 166.9, 158.3, 142.0, 138.2,
129.3, 129.1, 128.9, 97.0, 90.3, 85.0, 83.0 (d, J = 9.1 Hz), 77.2 (d, J =
157.0 Hz), 75.2, 72.1, 69.2 (d, J = 7.6 Hz), 64.3, 55.3, 37.3, 35.0 (d, J =
9.1 Hz), 32.6 (d, J = 6.0 Hz), 22.6. 31P NMR (162 MHz, D2O) δ
14.95, 0.36. HRMS (ESI) calcd for C24H33N4O13P2 [M − H]−,
647.1519; found, 647.1525.
Trisodium Cytidin-5′-yl-[(1R,3R,4S)-3-benzyloxy-4-acetamido-
methyl-1-cyclopentyl-phosphonatomethyl]-phosphate (6). This
compound was prepared in the same manner as described in the
preparation of 5S. Quantities: 47e (68.4 mg, 0.051 mmol), Pd/C
(136.8 mg, 10%), 1,4-cyclohexadiene (508.2 mg, 0.60 mL, 6.120
mmol, 97%), EtOH (2.2 mL), and eluent (H2O → MeOH/H2O =
1:1), affording crude product 6 (17.6 mg, 52% yield). The
diastereoisomers were further separated by preparative RP-HPLC,
converted to the form of sodium salt by ion-exchange (IR 120 Na+)
and lyophilized from water to give 6s and 6l as white powder. 6s: RP-
HPLC, column B, 0−16 min linear gradient 8−11.5% CH3CN, 100
mM NH4HCO3 buffer (pH = 8.4), 15 mL/min flow; tR = 12.65 min;
[α]3D0 = −3.6 (c = 0.2, H2O). 1H NMR (600 MHz, D2O) δ 8.01 (d, J =
7.8 Hz, 1H), 7.41−7.37 (m, 5H), 6.08 (d, J = 7.8 Hz, 1H), 5.93 (d, J =
3.6 Hz, 1H), 4.59 (d, J = 11.4 Hz, 1H), 4.46 (d, J = 11.4 Hz, 1H), 4.33
(t, J = 6.0 Hz,, 1H), 4.26−4.17 (m, 5H), 3.75 (dt, J = 8.4, 6.6 Hz, 1H),
3.20 (dd, J = 13.8, 7.2 Hz, 5H), 3.12 (dd, J = 13.8, 7.2 Hz, 1H), 2.47
(dt, J = 12.6, 6.6 Hz, 1H), 2.41 (brs, 1H), 2.18−2.12 (m, 1H), 1.99−
1.93 (m, 1H), 1.92 (s, 3H), 1.64 (td, J = 11.4, 9.0 Hz, 1H), 1.54 (ddd,
J = 12.6, 7.8, 4.8 Hz, 1H). 13C NMR (151 MHz, D2O) δ 174.7, 166.9,
158.3, 142.2, 138.2, 129.4, 129.3, 128.9, 97.1, 90.5, 84.1, 83.4 (d, J =
9.1 Hz), 79.0 (d, J = 149.5 Hz), 75.3, 71.9, 69.2, 64.5 (d, J = 4.5 Hz),
43.6, 43.1, 38.4, 35.1 (d, J = 4.5 Hz), 31.5 (d, J = 7.6 Hz), 22.6. 31P
NMR (162 MHz, D2O) δ 14.38, 0.70. HRMS (ESI) calcd for
C25H35N4O13P2 [M − H]−, 661.1676; found, 661.1675. 6l: RP-HPLC,
column B, 0−16 min linear gradient 8−11.5% CH3CN, 100 mM
P
J. Med. Chem. XXXX, XXX, XXX−XXX