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C.O.R. Junior et al. / European Journal of Medicinal Chemistry 85 (2014) 615e620
Table 1
Nitric oxide production and cellular viability of J774 A.1 stimulated by LPS/IFN-
g
and treated with genistein derivatives.a
Cellular viabilityd (%)
6.5c
Compoundsb
NO production (
mM)
6.5c
13c
26c
0.7 0.1*
13c
85.5 1.4
26c
30.3 17.1
6
7
8
9
17.7 2.3:
9.8 1.5*,:
9.7 0.5*,:
12.7 1.5*
11.5 0.4*,:
7.2 0.6*,:
8.7 0.6*,:
13.4 0.4*
12.8 0.7*,:
11.8 0.9*,:
10.9 1.7*,:
13.1 0.8*
102.1 4.3
89.7 2.1
93.6 1.9
94.7 2.8
82.1 8.3
68.1 4.9
99.9 0.4
111.7 17.7
12.3 0.7*,:
9.1 0.6*,:
10.4 2.1*,:
11.7 0.2*,:
14.3 0.6*,:
8.5 0.3*,:
14.5 0.6*
87.3 0.9
95.2 1.6
92.7 1.1
86.7 5.8
64.3 6.9
96.6 2.1
103.9 2.9
83.6 8.1
88.9 2.1
87.4 1.5
87.9 2.0
44.9 0.1
94.8 0.4
110.1 1.4
10
11
12
Genistein
12.1 0.2*
11.3 0.5*,:
9.1 0.2*,:
13.8 0.6*
*p < 0.05 to NO production by cells treated versus cells stimulated not treated, (NO production ¼ 16.5 1.4
mM).
:
p < 0.05 of genistein versus genistein derivative.
a
The results represent at least three independent experiments and are presented as the mean SEM.
b
(6): 7-O-(100,200:300,400-di-O-isopropylidene-600-deoxy-
a-
D
-galactopyrane-600-yl)-genistein, (7): 7,40-di-O-(100,200:300,400-di-O-isopropylidene-600-deoxy-
a
-
D
-galactopyrane-600-yl)-
genistein, (8): 7-O-(600-deoxy-
methyl-200,300,400-tri-O-acetyl-600-deoxy-
deoxy-
-glicopyrane-600yl)-genistein.
Concentration of compounds ( M).
Not treated cells viability 100 1.1%.
a-D
-galactopyrane-600-yl)-genistein, (9): 7-O-(100-O-methyl-200,300,400-tri-O-acetyl-600-deoxy-
a-D
-glicopyrane-600-yl)-genistein, (10): 7,40-di-O-(100-O-
a-D
-glicopyrane-600-yl)-genistein, (11): 7-O-(100-O-methyl-600-deoxy- -glicopyrane-600-yl)-genistein, (12): 7,40-di-O-(100-O-methyl-600-
a-D
a-D
c
m
d
Table 2
Nitric oxide production and cellular viability of RAW 264.7 stimulated by LPS/IFN-
g
and treated with genistein derivatives.a
Compoundsb
NO production (
6.5c
m
M)
Cellular viabilityd (%)
6.5c
13c
26c
13c
88.1 4.5
89.4 6.2
98.2 0.8
103.5 2.6
98.4 2.4
88.1 2.4
99.2 1.2
102.9 5.2
26c
71.3 5.5
94.4 2.7
95.8 2.8
102.8 1.2
101.3 0.8
78.3 2.8
92.2 2.5
107.9 2.8
6
7
8
9
2.4 0.3*,:
2.6 1.2:
1.4 1.1*,:
2.1 0.1*,:
1.9 0.3*,:
1.4 0.4*,:
0.0 0.6*,:
6.3 0.8*
2.5 0.2*,:
2.7 1.0:
0.0 0.4*,:
4.2 0.2:
2.5 0.3*,:
1.0 0.1*,:
0.0 1.0*,:
6.3 0.7*
3.1 0.4:
2.4 1.0*,:
0.0 0.4*,:
2.4 0.4*,:
4.1 0.3:
1.7 0.5*,:
0.0 1.0*,:
6.1 0.4*
87.6 1.3
98.9 1.9
95.9 2.9
103.0 3.2
102.8 0.8
91.3 1.0
93.6 1.1
128.3 13.8
10
11
12
Genistein
*p < 0.05 of NO production by cells treated versus cells stimulated not treated, (NO production ¼ 6.4 0.5
mM).
:
p < 0.05 of genistein versus genistein derivative.
a
The results represent at least three independent experiments and are presented as the mean SEM.
b
(6): 7-O-(100,200:300,400-di-O-isopropylidene-600-deoxy-
a-
D
-galactopyrane-600-yl)-genistein, (7): 7,40-di-O-(100,200:300,400-di-O-isopropylidene-600-deoxy-
a
-
D
-galactopyrane-600-yl)-
genistein, (8): 7-O-(600-deoxy-
methyl-200,300,400-tri-O-acetyl-600-deoxy-
deoxy-
-glicopyrane-600yl)-genistein.
Concentration of Compounds ( M).
Not treated cells viability 100 1.8%.
a-D
-galactopyrane-600-yl)-genistein, (9): 7-O-(100-O-methyl-200,300,400-tri-O-acetyl-600-deoxy-
a-D
-glicopyrane-600-yl)-genistein, (10): 7,40-di-O-(100-O-
a-D
-glicopyrane-600-yl)-genistein, (11): 7-O-(100-O-methyl-600-deoxy- -glicopyrane-600-yl)-genistein, (12): 7,40-di-O-(100-O-methyl-600-
a-D
a-D
c
m
d
4.3. Synthesis of 7-O-(600-deoxy-
genistein (8)
a
-D
-galactopyrane-600-yl)-
mixture was stirred for 1 h, and methyl 2,3,4-tri-O-acetyl-6-deoxy-
6-iodo- -glucopyranoside (5, 2.25 mmol) was added to the so-
a-D
lution. The mixture was stirred at 80 ꢀC for 48 h, DMF was removed
under reduced pressure, and the residue was extracted with diethyl
ether and water. The organic phase was dried with sodium sulfate,
and the solvent was concentrated under reduced pressure. The
residue was purified by column chromatography (hexane/AcOEt) to
afford 9 and 10.
One milliliter of trifluoroacetic acid at room temperature was
added to a solution of 6 (0.2 mmol) in 2 mL of CH2Cl2 and 1 mL of
H2O. The mixture was stirred at room temperature for 24 h and
evaporated under reduced pressure. The residue was purified by
column chromatography (CH2Cl2/MeOH) to afford 8 (70% yield).
mp: 175e178 ꢀC; 1H NMR (300 MHz, C5D5N):
d
4.24e4.40 (m,
9 (55% yield); mp: 89e92 ꢀC; [
NMR (300 MHz, CDCl3): d 2.00 (s, 3H), 2.02 (s, 3H), 2.06 (s, 3H), 3.42
a
]
D
¼ þ108 (c 0.1, CH2Cl2); 1H
2H), 4.55e4.84 (m, 4H), 5.05 (s, 0.5H), 5.34 (d, 0.5H, J ¼ 7.3 Hz), 6.15
(br s, OHaliphatic), 6.69 (s, 1H), 6.74 (s, 1H), 7.30 (d, 2H, J ¼ 8.4 Hz),
7.72 (d, 2H, J ¼ 8.4 Hz), 8.18 (s, 1H), and 13.56 (s, 1H); 13C NMR
(s, 3H), 4.06e4.14 (m, 3H), 4.88e4.97 (m, 2H), 5.14 (t,1H, J ¼ 9.0 Hz),
5.51 (t,1H, J ¼ 9.0 Hz), 6.32 (d,1H, J ¼ 3.0 Hz), 6.67 (d,1H, J ¼ 3.0 Hz),
6.84 (d, 2H, J ¼ 9.0 Hz), 7.31 (d, 2H, J ¼ 9.0 Hz), 7.81 (s, 1H) and 8.80
(75 MHz, C5D5N):
d 56.3, 66.0, 70.8, 71.0, 71.2, 71.7, 72.1, 72.3, 72.5,
75.3, 76.5, 85.2, 94.4, 95.8, 100.6, 108.0, 117.6, 129.9, 130.7, 132.2,
155.0, 159.6, 160.6, 164.4, 166.7 and 182.6; IR (KBr): 3118e3380,
2921e2986,1669 and 1055 cmꢁ1; HRMS calculated for [MþH]þ was
433.1129, the value found was 433.1122.
(s, 1H); 13C NMR (75 MHz, CDCl3):
d 20.9, 55.8, 67.4, 67.6, 69.4, 70.3,
71.0, 93.3, 96.9, 98.7, 106.8, 115.8, 122.0, 122.6, 124.0, 130.4, 153.0,
156.7, 158.0, 162.9, 164.3, 170.1, 170.4 and 181.1; IR (KBr):
3154e3446, 2852e2956, 1751, 1656 and 1045 cmꢁ1; HRMS calcu-
lated for [MþNa]þ was 595.1430, the value found was 595.1409.
10 (20% yield); mp: 102e103 ꢀC; [
NMR (300 MHz, CDCl3): d 1.99 (s, 6H), 2.01 (s, 6H), 2.06 (s, 6H), 3.42
a
]
D
¼ þ28 (c 0.1, CH2Cl2); 1H
4.4. Synthesis of 7-O-(100-O-methyl-200,300,400-tri-O-acetyl-600-deoxy-
a-D
-glicopyrane-600-yl)-genistein (9) and 7,40-di-O-(100-O-methyl-
200,300,400-tri-O-acetyl-600-deoxy- -glicopyrane-600-yl)-genistein (10)
a-D
(s, 6H), 4.04e4.15 (m, 6H), 4.89e4.96 (m, 4H), 5.15 (m, 2H), 5.51 (t,
2H, J ¼ 9.0 Hz), 6.33 (d, 1H, J ¼ 3.0 Hz), 6.38 (d, 1H, J ¼ 3.0 Hz), 6.94
(d, 2H, J ¼ 9.0 Hz), 7.41 (d, 2H, J ¼ 9.0 Hz), 7.84 (s, 1H) and 8.81 (s,
Potassium carbonate (1.33 mmol) at room temperature was
added to a solution of genistein 1 (0.75 mmol) in 10 mL of DMF. The
1H); 13C NMR (75 MHz, CDCl3):
d 20.9, 55.7, 67.2, 67.4, 67.6, 67.9,