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H.-P. Wei et al. / European Journal of Medicinal Chemistry 44 (2009) 665e669
(3H, m, 10-H, 20-H, 40-H), 5.17e5.13 (2H, d, 50-OH, 20-OH),
24
21
18
15
12
9
3.71e3.67 (2H, d, 6-NH2), 2.29e2.20 (2H, m, 50-CH2), 1.92
(2H, m, 30-CH2); Anal. calcd for C10H13N5O3: C 47.81, H
5.22, N 27.88, O 19.10; found: C 47.75, H 5.16, N27.901,
O19.16.
compound 1
compound 2
compound 3
compound 4
compound 5
Compound 2 was obtained as white flakes from H2O; mp
177e179 ꢁC; Rf 0.37, silica gel-GF254
, EtOAc/MeOH/
NH4OH (99:10:1); Yield 75.5%; UV (MeOH) lmax (log e)
213 (4.05), 260 (3.87) nm; IR (KBr)nmax 3378, 2933, 1741,
1632, 1544, 1478, 1375, 1287, 1099, 839, 722 cmꢂ1 1H
;
NMR (CDCl3, 300 MHz) d 8.62 (1H, m, 6-NHeCOe), 8.44
(1H, s, 2-H), 8.23 (1H, s, 8-H), 5.83e5.65 (3H, m, 10-H, 20-
H, 40-H), 4.36e4.23 (2H, m, 50-OH, 20-OH), 2.39e2.29 (4H,
m, 30-CH2, 50-CH2), 1.59 (2H, m, eCH2), 1.17e1.12 (3H, t,
eCH3, J ¼ 7 Hz); Anal. calcd for C13H17N5O4: C 50.81, H
5.58, N 22.79, O 20.82; found: C 51.05, H 5.51, N 22.81, O
19.95.
6
3
0
0
1
2
3
4
5
6
7
Time (h)
8
9
10 11 12 13 14 15
Compound 3 was obtained as white flakes from EtOAc; mp
139e141 ꢁC; Rf 0.45, silica gel-GF254, EtOAc/MeOH/NH4OH
(99:10:1); Yield 64.3%; UV (MeOH)lmax (log e) 213 (4.12),
260 (3.68) nm; IR (KBr)nmax 3323, 2928, 1742, 1632, 1546,
Fig. 2. Mean blood concentrationetime profiles of transformed cordycepin
from compounds 2e5 as compared with compound 1 after oral administrations
(500 mmol/kg), each point represents the arithmetic mean (n ¼ 5).
1
1456, 1380, 1287, 1096, 842, 722 cmꢂ1; H NMR (CDCl3,
300 MHz) d 8.60 (1H, m, 6-NHeCOe), 8.22 (1H, s, 2-H),
8.01 (1H, s, 8-H), 5.92e5.86 (3H, m, 10-H, 20-H, 40-H),
4.39e4.26 (2H, m, 50-OH, 20-OH), 2.32e1.23 (16H, m, e
CH2e), 0.87e0.85 (3H, t, eCH3, J ¼ 8 Hz); Anal. calcd for
C18H27N5O4: C 57.28, H 7.21, N 16.96, O 18.56; found: C
57.37, H 7.17, N 17.03, O 18.48.
dehydrated pyridine at 60e70 ꢁC under magnetic stirring and ni-
trogen protection. After 10 h of reflux, the reaction solution was
concentrated and dissolved in 50 mL EtOAc with 5% HAc, then
purified by a silica gel column (3 ꢃ 75 cm, 100 mL of 200e300
mesh silica gel, the eluent was 400 mL EtOAc with 5% HAc, fol-
lowed by 200 mL MeOH). The resulting eluent was concentrated
and crystallized at 0 ꢁC to get the corresponding products.
Compound 1 was obtained as white needles or flakes from
H2O; mp 229e230 ꢁC; Rf 0.25, silica gel-GF254, EtOAc/
MeOH/NH4OH (99:10:1); UV (MeOH) lmax (log e) 213
(3.87), 260 (4.09) nm; IR (KBr)nmax 3332, 3142, 1672,
1608, 1479, 1420, 1385, 1340, 1307, 1209, 1110, 1059,
Compound 4 was obtained as ivory white needles or flakes
from EtOAc; mp 105e106 ꢁC; Rf 0.67, silica gel-GF254
,
EtOAc/MeOH/NH4OH (99:10:1); Yield 45.7%; UV
(MeOH)lmax (log e) 213 (4.16), 261 (3.87) nm; IR (KBr)nmax
3396, 2935, 1737, 1632, 1544, 1459, 1378, 1288, 1098, 899,
1
722 cmꢂ1; H NMR (CDCl3, 300 MHz) d 8.73 (1H, m, 6-
NHeCOe), 8.34 (1H, s, 2-H), 8.19 (1H, s, 8-H), 5.87e5.66
(3H, m, 10-H, 20-H, 40-H), 4.39e4.33 (2H, m, 50-OH, 20-
OH), 2.36e1.25 (24H, m, eCH2e), 0.87e0.85 (3H, t, e
CH3, J ¼ 6 Hz); Anal. calcd for C22H35N5O4: C 60.95, H
8.14, N 16.15, O 14.76; found: C 60.88, H 8.11, N 16.05, O
14.55.
1081, 1038, l008, 727 cmꢂ1
;
300 MHz) d 8.37(1H, s, 2-H), 8.15(1H, s, 8-H), 5.96e5.86
1H NMR (DMSO-d6,
90
80
70
60
50
40
30
20
10
0
Compound 5 was obtained as ivory white flakes from
EtOAc; mp 101e103 ꢁC; Rf 0.68, silica gel-GF254, EtOAc/
MeOH/NH4OH (99:10:1); Yield 34.5%; UV (MeOH)lmax
(log e) 212 (4.07), 261 (3.98) nm; IR (KBr)nmax 3424, 2924,
2854, 1745, 1632, 1545, 1454, 1374, 1284, 1096, 795,
721 cmꢂ1 1H NMR (CDCl3, 300 MHz) d 8.66e8.64 (1H,
;
m, 6-NHeCOe), 8.31 (1H, s, 2-H), 8.06 (1H, s, 8-H),
5.91e5.85 (3H, m, 10-H, 20-H, 40-H), 4.39e4.22 (2H, m, 50-
OH, 20-OH), 2.36e1.23 (36H, m, eCH2e), 0.90e0.85 (3H,
t, eCH3, J ¼ 7.5 Hz); Anal. calcd for C28H47N5O4: C 64.96,
H 9.15, N 13.53, O 12.36; found: C 64.55, H 9.11, N 13.50,
O 12.29.
6.3. HPLC analysis
0
2
4
6
8
10
12
14
16
18
Cn
To determine the recovery of compounds 1e5 from mice
blood, 0.5, 1.0 or 1.5 mmol/L of standard compounds 1e5
Fig. 3. Scatter diagram of AUC and Cn.