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+ AH SOOH + A
Stage II :
A
+ X
non radical materials
Where: S: is the oxidized substance.
AH: is the phenolic antioxidant.
A
:
is another radical species or the same as A
.
Chart i
Although the first stage is a reversible process, the second stage is irreversible and
must produce stable radical terminated compounds. During the course of the
antioxidant mechanism study of curcumin, it was shown that dimerization was a main
termination process of the radical reaction of curcumin itself
[35]
. On the other hand in
the food system, the antioxidant coexists with a large amount of oxidizable
biomolecules such as poly- unsaturated lipids. These biomolecules produce reactive
peroxyl radicals during their oxidation, which may act as X
C (reported
m.p. = 181
-183
C
[65]
, 184
-185
C
[107]
, 184
-186
C
[122]
, 180
-182
C
[126]
, 175
-177
C
[136]
, 182
-183
C
[148]
). IR of curcumin (1) (cm
-1
): 3468.5 (br, OH), 1627.8 (C=O),
1604.4, 1508.1 (C=C, Ar), 1282.4, 1027.3 (
as
and
s
C-O-C);
1
H-NMR of 1 (DMSO-
103
d
6
) (ppm) (300 MHz) (figure 1): 3.8366 (s, 6H, 2 x OCH
3
), 6.0618 (s, 1H, H
a
),
6.711 (d, 2H, 2 x H
b
, J = 16.6 Hz), 6.8227 (d, 2H, 2 x H
A
, J
AB
= 8.22 Hz, ortho
coupling), 7.1569 (d, 2H, 2 x H
B
, J
AB
= 8.03 Hz, ortho coupling), 7.3273 (s, 2H, 2 x
H
X
), 7.5470 (d, 2H, 2 x H
c
, J = 15.35 Hz), 9.7411 (s, 2H, 2 x phenolic OH), 10.1312
(s, 1H, enol OH);
13
C-NMR of 1 (DMSO-d
6
) ppm (300 MHz): 56.2179 (2 x OCH
3
),
101.4313 (C-4), 111.8205 (2 x C-2'), 116.2327 (2 x C-5'), 121.6219 (2 x C-6'),
123.7059 ( 2 x C-1'), 126.8662 (C-2 + C-6), 141.2783 (C-1 + C-7), 148.5301 (2 x C-
3'), 149.8889 (2 x C-4'), 183.7664 (C-3 + C-5).
Ethyl curcumin (2) was obtained from 4-hydroxy-3-ethoxy- benzaldehyde in 68%
yield; m.p. = 143
-160
C)
[138]
. IR of ethyl curcumin (2) (cm
-
1
): ;
1
H-NMR of 2 (DMSO-d
6
) ppm (500 MHz) (figure 2): 1.358 (t, 6H, 2 x
OCH
2
CH
3
, J = 7 Hz), 4.094 (q, 4H, 2 x CH
2
CH
3
, J = 7 Hz), 6.050 (s, 2H, H
a
), 6.746
(d, 2H, 2 x H
b
, J = 16 Hz), 6.850 (d, 2H, 2 x H
A
, J
AB
= 8 Hz, ortho coupling), 7.1495
(d, 2H, 2 x H
B
, J
AB
= 8.5 Hz, ortho coupling), 7.305 (s, 2H, 2 x H
X
), 7.552 (d, 2H, 2 x
H
c
, J = 16 Hz), 9.607 (s, 2H, 2 x phenolic OH);
13
C-NMR of 2 (DMSO-d
6
) ppm
(500 MHz) (figure 3): 15.16 (2 x OCH
2
CH
3
), 64.36 (2 x OCH
2
CH
3
), 101.37 (C-4),
112.88 (2 x C-2'), 116.25 (2 x C-5'), 121.49 (2 x C-6'), 123.47 (2 x C-1'), 126.79 (C-2
+ C-6), 141.20 (C-1 + C-7), 147.59 (2 x C-3'), 150.05 (2 x C-4'), 183.67 (C-3 + C-5);
MS of 2 (figure 4) m/z (% relative abundance): M
+
396 (23), 378 (17), 349 (11), 300
(8), 246 (10), 231 (14), 231 (14), 217 (11), 206 (11), 205 (26), 204 (47), 191 (70), 178
(11), 177 (43), 176 (42), 165 (15), 163 (78), 162 (9), 159 (24), 148 (17), 147 (47), 146
(16), 145 (69), 143 (8), 136 (41), 134 (39), 131 (56), 123 (38), 119 (14), 117 (48), 115
(24), 110 (30), 107 (33), 105 (22), 103 (44), 95 (7), 94 (7), 91 (65), 90 (20), 89 (100),
81 (12), 79 (37), 78 (41), 77 (88), 69 (41), 67 (12), 66 (13), 65 (57), 43 (57).
104
5.2.2. Di-O-Chloroacetylcurcumin (3a) and di-O-chloropropionyl-
curcumin (3b)
O O
H
3
CO
O
OCH
3
O
O
(CH
2
)n
Cl
Cl
(CH
2
)n
O
(3a) n = 1 (3b) n = 2
5.2.2.1. Method A
Sodium carbonate (2 g) was added to a stirred ice cold solution of curcumin (1)
(3.68g, 0.01 mol) in dry acetone (50 ml) for 15 min. Chloroacetyl chloride or
chloropropionyl chloride (0.025 mol) was dropwise added and the mixture was stirred
for 3 days at RT. The reaction mixture was filtered, evaporated and extracted with
EtOAc (3 x 30 ml). The organic layer was dried over anhydrous MgSO
4
(5 g) and
evaporated under reduced pressure to give an orange oil. Addition of EtOH (20 ml)
gave a yellow precipitate which was filtered off. The product was purified by column
chromatography using toluene/MeOH (97.5: 2.5 v/v) as eluent (Table 1).
IR of di-O-chloroacetylcurcumin (3a) (cm
-1
): 3414.8 (OH, intramolecularly H-
bonded), 1768.7 (C=O, ester), 1635.7 (C=O, ketone), 1599.9, 1507.6 (C=C Ar),
1254.8, 1122.4 (
as
and
s
C-O-C).
1
H-NMR of 3a (CDCl
3
) ppm (300 MHz) (figure
5): 3.876 (s, 6H, 2 x OCH
3
), 4.1003 (s, 4H, 2 x CH
2
), 5.8557(s, 2H, H
a
), 6.56905 (d,
dist, 2H, 2H
b
, J = 15.9 Hz), 7.0754-7.2613 (m, 6H, 2 x 3 Ar-H), 7.6116 (d, dist, 2H,
2H
c
, J = 15.7 Hz).
IR of di-O-chloropropionylcurcumin (3b) (cm
-1
): 3414.2 (OH, intramolecularly
H-bonded), 1746.6, (C=O, ester), 1635.1 (C=O, ketone), 1607.4, 1506.4 (C=C Ar),
1253, 1129.3 (
as
and
s
C-O-C).
1
H-NMR of 3b (DMSO-d
6
) ppm (300 MHz):
105
3.1223 (t, 4H, 2 x CH
2
-Cl, J = 6.1 Hz), 3.8403 (s, 6H, 2 x OCH
3
), 3.8952 (t, 4H, 2 x
COCH
2
, J = 6.33 Hz), 6.2156 (s, 2H, H
a
), 7.0095 (d, dist, 2H, 2 x H
b
, J = 15.9 Hz),
7.1524-7.5351 (m, 6H, 2 x 3 Ar-H), 7.6569 (d, dist, 2H, 2 x H
c
, J = 15.9 Hz); MS of
3b: m/z (% relative abundance): M
+
549.5 (absent), 284 (14.28), 241 (8.92), 227
(3.57), 199 (4.46), 185 (16.07), 171 (6.25), 143 (7.14), 129 (37.5), 111 (14.28), 97
(31.25), 83 (37.5), 73 (100), 69 (57.25).
Table 1: Physicochemical Data of di-O-chloroacetylcurcumin (3a) and
di-O-chloropropionylcurcumin (3b)
M. wt Mol. Formula
m.p.
C
Yield
(%)
Structure
Cpd
No
521
C
25
H
22
Cl
2
O
8
136
43
O O
H3CO OCH3
OCOCH2Cl ClH2COCO
3a
549
C
27
H
26
Cl
2
O
8
143
38
O O
H3CO OCH3
OCOCH2CH2Cl ClH2CH2COCO
3b
5.2.2.2. Method B
a) 3-Alkoxy-4-chloroacyloxybenzaldehyde (4a-d)
CHO
O
O
(CH
2
)n
Cl
R
1
O
(4a) R
1
= CH
3
n = 1; (4b) R
1
= CH
3
n = 2
(4c) R
1
= C
2
H
5
n = 1; (4d) R
1
= C
2
H
5
n = 2
a.1. The appropriate acid chloride (0.01 mol) in chloroform (20 ml) was added
dropwise over a period of 5 min to an ice-cold stirred solution of the aldehyde (0.01
106
mol) and 1N NaOH (2 ml) in chloroform (5 ml). The mixture was stirred at RT for 1
day and then evaporated to dryness. The residue was extracted (3 x 20 ml) with
EtOAc and 0.1 N NaOH. The combined organic layers were washed with water (3 x
20 ml), dried over anhydrous MgSO
4
(5 g) and removed under reduced pressure to
give a colorless oil. Trituration of the crude product with EtOH (20 ml) gave colorless
crystals of 4a-d which were crystallized from EtOH (Table 2).
a.2. The selected acid chloride (0.01 mol) was dropwise added to an ice cold stirred
solution of the appropriate aldehyde (0.01mol) in dry acetone and anhydrous K
2
CO
3
.
The mixture was further stirred in an ice-bath for 3 h, filtered and concentrated.
Dilution with EtOH (20 ml) precipitated colorless crystals of 4a-d which were filtered
and crystallized from EtOH (Table 2).
IR of 4a (cm
-1
): 2933.4, 2847 (CHO), 1765 (C=O ester), 1698 (C=O aldehyde),
1600, 1505 (C=C aromatic), 1180, 1150, 1100, 1020 (
as
and
s
C-O-C);
1
H- NMR of
4a (DMSO-d
6
) ppm (500 MHz) (figure 6): 3.8445 (s, 3H, OCH
3
), 4.7157 (s, 2H,
CH
2
-Cl), 7.3897 (d, 1H, H
A
, J
AB
= 8.40, ortho coupling), 7.5647, 7.5800 (dd, 1 H, H
B
,
J = 1.55 Hz and 7.65 Hz, meta and ortho coupling), 7.6075 (d, 1H, H
X
, J
AX
= 1.55 Hz,
para coupling), 9.9476 (s, 1H, CHO).
IR of 4b (cm
-1
): 3073, 2940 (CHO), 1760 (C=O ester), 1698 (C=O aldehyde),
1601, 1495 (C=C aromatic), 1154, 1120, 1068, 1025 (
as
and
s
C-O-C).
1
H- NMR of 4c (DMSO-d
6
) ppm (300 MHz) (figure 7): 1.3138 (t, 3H,
OCH
2
CH
3
, J = 6.87 Hz), 4.15575 (q, 2H, OCH
2
CH
3
, J = 6.87 Hz), 4.7413 (s, 2H,
CH
2
Cl), 7.4136 (d, 1H, H
A
, J
AB
= 7.98 Hz, ortho coupling), 7.5762, 7.6051 (dd, 1H,
H
B
, J = 1.65 Hz and 8.07 Hz, meta and ortho coupling), 7.6285 (d, 1H, H
X
, J
AX
= 1.63
Hz, para coupling), 9.9709 (s, 1H, CHO).
107
Table 2: Physicochemical Data of the Synthesized Compounds 4a-d
Cpd
No
Structure
Method
A
Reaction
Time(h)
Method
B
Reaction
Time(h)
Yield
%
A
Yield
%
B
m.p.
C
Molecular
Formula
M.wt
4a
OCH
3
CHO
ClH
2
COCO
3
24
43
78
68-
71
C
10
H
9
ClO
4
228.5
4b
OCH
3
CHO
ClH
2
CH
2
COCO
24
3
41
72
65-
69
C
11
H
11
ClO
4
242.5
4c
OCH
2
CH
3
CHO
ClH
2
COCO
24
3
42
67
61-
64
C
11
H
11
ClO
4
242.5
4d
OCH
2
CH
3
CHO
ClH
2
CH
2
COCO
24
3
39
61
55-
59
C
12
H
13
ClO
4
256.5
b) Di-O-Chloroacylcurcumin (3a,b) and di-O-chloroacylethyl
curcumin (3c,d)
O
OR
1
O
R
1
O
O O
(CH
2
)n n(H
2
C)
O O
Cl
Cl
(3a-d)
To a stirred solution of the appropriate 3-alkoxy-4-chloroacyloxybenzaldehyde
(4a-d) (0.04 mol) in dry EtOAc (20 ml) was added tri-(n-butyl) borate (21 ml, 0.08
mol) and the mixture was stirred at RT for 10 min. The previously prepared complex
formed by stirring for 1 h acetylacetone (2 g, 0.02 mol) with boric anhydride (1 g,
108
0.014 mol) was then added to this solution. After stirring for 5 min at RT, n-
butylamine (0.1 ml) was dropwise added every 10 min (total amount 0.4 ml) and the
reaction mixture was stirred at RT for an overnight. 0.4 N HCl (30 ml, 60
C) was then
added and the mixture was further stirred for 2 h followed by extraction with EtOAc
(3 x 25 ml). The combined organic layers were washed with H
2
O, dried over
anhydrous MgSO
4
(5 g) and concentrated to ~ 15 ml. MeOH (15 ml) was added and
the mixture was allowed to stand in the refrigerator for 3 h to give a yellow
precipitate of 3a-d which was filtered off, washed with cold MeOH and dried (Table
3).
IR and
1
H-NMR spectra of compounds 3a,b were coinciding with those obtained
from method A.
IR of 3c (cm
-1
): 3435.6 (OH, intramolecularly H-bonded), 1746 (C=O, ester),
1630.9 (C=O, ketone), 1558.1, 1496.3 (C=C aromatic), 1270, 1157 (
as
and
s
C-O-
C);
1
H- NMR of 3c (CDCl
3
) ppm (300 MHz) (figure 8): 1.4164 (t, 6H, 2 x
OCH
2
CH
3
, J = 6.87 Hz), 4.1049 (q, 4H, 2 x OCH
2
CH
3
, J = 6.87 Hz), 4.3329 (s, 4H, 2
x CH
2
-Cl), 5.8457(s, 2H, H
a
), 6.5553 (d, 2H, 2 x H
b
, J = 15.93 Hz), 7.0764-7.1762
(m, 6H, 2 x 3 Ar-H), 7.605 (d, 2H, 2 x H
c
, J = 15.93 Hz); MS of 3c (figure 9) m/z (%
relative abundance): M
+
548 (0.24), M
+
+2 550 (0.21), M
+
+4 552, 533 (0.23), 531
(0.28), 529 (0.29), 482 (0.24), 471 (0.21), 412 (0.15), 377 (0.13), 373 (0.3), 343
(0.21), 323 (0.12), 296 (0.14), 274 (0.14), 244 (0.26), 242 (0.6), 227 (0.15), 216
(0.24), 191 (0.17), 166 (2), 137 (2), 120 (2), 87 (12), 85 (69), 83 (100).
1
H- NMR of 3d (CDCl
3
) ppm (500 MHz) (figure 10): 1.442 (t, 6H, 2 x
OCH
2
CH
3
, J = 7 Hz), 3.105 (t, 4H, 2 x COCH
2
, J = 7Hz), 3.904 (t, 4H, 2 x CH
2
-Cl, J
= 7 Hz), 4.131 (q, 4H, 2 x OCH
2
CH
3
, J = 7 Hz), 5.871 (s, 1H, H
a
), 6.577 (d, 2H, 2 x
H
b
, J = 16 Hz), 7.098 (d, 2H, 2 x H
A
, J
AB
= 8 Hz, ortho coupling), 7.1735 (d, 2H, 2 x
109
H
B
, J
AB
= 8.5 Hz, ortho coupling), 7.286 (s, 2H, 2 x H
X
), 7.559 (d, 2H, 2 x H
c
, J = 16
Hz), 9.965 (s, 1H, enol OH).
Table 3: Physicochemical Data of the Synthesized Compounds 3a-d
O
OR
1
O
R
1
O
O O
(CH
2
)n n(H
2
C)
O O
Cl
Cl
(3a-d)
M. wt M. Formula m.p.
C
Yield
%
n R
1
No.
521
C
25
H
22
Cl
2
O
8
136
61
1
CH
3
3a
549
C
27
H
26
Cl
2
O
8
143
56
2
CH
3
3b
549
C
27
H
26
Cl
2
O
8
130
59
1
C
2
H
5
3c
577
C
29
H
30
Cl
2
O
8
145
54
2
C
2
H
5
3d
5.2.3. 2-Chloroethylamine monohydrochloride and bis(2-chloroethyl)amine
hydrochloride
NH
2
CH
2
CH
2
Cl . HCl
NH
CH
2
CH
2
Cl
CH
2
CH
2
Cl
. HCl
Thionyl chloride (30 ml) was added dropwise to a solution of ethanolamine or
diethanolamine (10 ml) in dry benzene (70 ml) in an ice bath. The mixture was heated
under reflux for h and cooled to RT. The oily residue obtained was separated and
crystallized from EtOH and diethyl ether (charcoal). m.p. = 143-146
C as reported
[160]
110
for 2-chloroethylamine monohydrochloride and 212-214
C as reported
[160]
for bis(2-
chloroethyl)amine hydrochloride.
5.2.4. 1,7-Bis(4-Alkyl(cycloalkyl or heteroaryl)aminoacyloxy)-3-
(methoxyphenyl)-
1,6-heptadiene-3,5-dione (5a-n) and 1,7-bis(4-alkyl (cycloalkyl or
heteroaryl)-
aminoacyloxy)-3-(ethoxyphenyl)-1,6-hepta- diene-3,5-dione (6a-n)
OR
1
O
R
1
O
O O
n(H
2
C)
O O
(CH
2
)n
R
3
R
2
N
NR
2
R
3
(5a-n, 6a-n)
Triethylamine (5 drops) were added to a solution of di-O-chloroacylcurcumin
3a,b or di-O-chloroacylethyl curcumin 3c,d (0.005 mol) in EtOH (30 ml). The
appropriate amine (0.01 mol) was then added and the mixture was heated under reflux
for the specified time (Table 4). EtOH was evaporated under reduced pressure and the
residue was crystallized from CHCl
3
(15ml) giving compounds 5a-n and 6a-n (Table
4).
1
H-NMR of 5a (DMSO-d
6
) ppm (300 MHz) (figure 11): 1.1609-1.9951 (hump,
br, 30H, 2 x Ad-H), 2.0858 (s, 4H, 2 x CO-CH
2
), 3.8311 (s, 6H, 2 x OCH
3
), 6.0334 (s,
br, 2H, H
a
), 6.7444 (d, 2H, 2x H
b
, J = 15.66 Hz), 6.8016 (d, 2H, 2 x H
A
, J
AB
= 8.25
Hz, ortho coupling), 7.1295, 7.1569 (dd, dist, 2H, 2 x H
B
, J = 1.5 Hz and 8.22 Hz,
meta and ortho coupling), 7.3099 (s, dist, 2H, 2 x NH), 7.1734 ( d, 2H, 2 x H
X
, J
AX
=
0.24 Hz, para coupling), 7.5315 (d, 2H, 2 x H
c
, J = 15.66 Hz); MS of 5a m/z (%
abundance): M
+
750 (absent), 135 (100), 107 (33.03), 93 (35.71), 83 (30.35), 81
(39.28), 79 (42.85) 77 (35.71).
111
IR of 5b (cm
-1
): 3550, 3414.4 (NH + OH intramolecular H-bonded), 1733.7
(C=O, ester), 1637.4 (C=O, ketone), 1618 (C=N mixed with C=C aromatic), 1510.2
(C=C, aromatic), 1272.1 1122.9 (
as
and
s
C-O-C);
1
H-NMR of 5b (DMSO-d
6
)
ppm (300 MHz): 2.0876 (s, 4H, 2 x CH
2
), 3.8384 (s, 12H, 4 x OCH
3
), 6.0599 (s, 2H,
H
a
), 6.7646 (d, 2H, 2 x H
b
, J = 15.66 Hz), 6.8236 (d, 2H, 2 x H
A
, J
AB
= 8.25 Hz, ortho
coupling), 7.1413, 7.1688 (dd, 2H, 2 x H
B
, J = 1.65 Hz and 8.25 Hz, meta and ortho
coupling), 7.32815 (d, 2H, H
X
, J
AX
= 1.65 Hz, para coupling), 7.5466 (d, 2H, 2 x H
c
,
J = 15.66 Hz, overlapping with benzothiazole multiplet), 7.525-7.610 (m, 6H, 2 x 3
benzothiazole-H), 9.6925 (s, 2H, 2 x NH, D
2
O-exchangeable); MS of 5b m/z (%
abundance): M
+
808 (absent), 163 (53.57), 161 (35.71), 127 (41.07), 125 (100), 97
(62.5), 83 (37.5), 79 (67.85), 77 (71.42).
IR of 5c (cm
-1
): 3425 (NH + OH intramolecularly H-bonded), 1727.3 (C=O,
ester), 1630 (C=O, ketone mixed with C=N), 1575, 1506 (C=C aromatic), 1272.5,
1120.7 (
as
and
s
C-O-C);
1
H-NMR of 5c (DMSO-d
6
) ppm (300 MHz): 2.5033 (s,
4H, 2 x CH
2
), 3.3522 (s, 6H, 2 x CH
3
), 3.835 (s, 6H, 2 x OCH
3
), 6.0590 (s, 2H, H
a
),
6.7631 (d, 2H, 2 x H
b
, J = 15.93 Hz), 6.8218 (d, 2H, 2 x H
A
, J
AB
= 8.22 Hz, ortho
coupling), 7.1409, 7.1676 (dd, 2H, 2 x H
B
, J = 1.65 Hz and 8.01 Hz, meta and ortho
coupling), 7.3272 (d, 2H, 2 x H
X
, J
AX
= 1.65 Hz), 7.5457 (d, 2H, 2 x H
c
, J = 15.66
Hz), 9.6852 (s, 2H, 2 x NH, D
2
O-exchangeable).
1
H-NMR of 5d (CD
3
OD) ppm (500 MHz): 2.292 (s, 2H, 2 x NH), 2.305 (t, dist,
4H, 2 x CH
2
-N), 3.230 (t, 4H, 2 x CH
2
-Cl, J = 3.5 Hz), 3.331 (s, 4H, 2 x COCH
2
-
N), 3.918 (s, 6H, 2 x OCH
3
), 4.903 (s, 2H, 2 x H
a
), 6.643 (d, 2H, 2 x H
b
, J = 15.5 Hz),
6.839 (d, 2H, 2 x H
A
, J
AB
= 8 Hz, ortho coupling), 7.119 (d, 2H, 2 x H
B
, 7.7 Hz, ortho
coupling), 7.227 (s, 2H, 2 x H
X
), 7.584 (d, 2H, 2 x H
c
, J = 15.5 Hz).
112
IR of 5e (cm
-1
): 3414.8 (OH intramolecularly H- bonded), 1728.4 (C=O, ester),
1637.6 (C=O, ketone), 1618.1, 1511 (C=C aromatic), 1272.6, 1122.9 (
as
and
s
C-O-
C);
1
H-NMR of 5e (DMSO-d
6
) ppm (300 MHz) (figure 12): 2.0867 (s, 4H, 2 x
COCH
2
-N), 2.5042 (t, 8H, 4 x N-CH
2
CH
2
-Cl, J = 3.57 Hz), 3.4841 (t, 8H, 4 x
NCH
2
CH
2
-Cl, overlapping with solvent signal), 3.8384 (s, 6H, 2 x OCH
3
), 6.0599 (s,
1H, H
a
), 6.7718 (d, 2H, 2 x H
b
, J = 15.9 Hz), 6.8245 (d, 2H, 2 x H
A
, J
AB
= 8.25 Hz,
ortho coupling), 7.1409, 7.1688 (dd, 2H, 2 x H
B
, J = 1.65 Hz and 8.37 Hz, meta and
ortho coupling), 7.3277 (d, 2H, 2 x H
X
, J
AX
= 1.38 Hz, para coupling), 7.5736 (d, 2H,
2 x H
c
, J = 15.9 Hz), 9.6953 (s, br, 1H, enol OH).
1
H-NMR of 5f (DMSO-d
6
) ppm (300 MHz): 3.1748 (s, 12H, 2 x N(CH
3
)
2
),
3.5387 (t, dist, 4H, 2 x CH
2
-N), 3.6657 (t, 4H, 2 x CH
2
-NH, J = 5.49 Hz), 3.8012 (s,
dist, 2H, 2 x NH, overlapping with OCH
2
), 3.8343 (s, 4H, 2 x COCH
2
-N), 4.0602 (s,
6H, 2 x OCH
3
), 6.0595 (s, 1H, H
a
), 6.7586 (d, 2H, 2 x H
b
, J = 15.75 Hz), 6.8337 (d,
2H, 2 x H
A
, J
AB
= 8.07 Hz, ortho coupling), 7.1513 (d, 2H, 2 x H
B
, J = 8.07 Hz, ortho
coupling), 7.3235 (s, 2H, 2 x H
X
), 7.5427 (d, 2H, 2 x H
c
, J = 15.75 Hz), 9.7368 (s, 1H,
enol H).
1
H-NMR of 5h (CD
3
OD) ppm (500 MHz) (figure 13): 1.184-2.172 (hump, br,
30H, 2 x Ad-H), 2.631 (t, dist, 4H, 2 x CH
2
-N, J = 6.2 Hz), 3.064 (t, 4H, 2 x CO-
CH
2
, J = 6.5 Hz), 3.882 (s, 2H, 2 x NH overlapping with OCH
3
signal), 3.921 (s, 6H,
2 x OCH
3
), 4.936 (s, 2H, H
a
), 6.632 (d, 2H, 2x H
b
, J = 15.7 Hz), 6.835 (d, 2H, 2 x H
A
,
J
AB
= 8 Hz, ortho coupling), 7.111 (d, 2H, 2 x H
B
, J = 7.8 Hz, ortho coupling), 7.218 (
d, 2H, 2 x H
X
), 7.577 (d, 2H, 2 x H
c
, J = 15.8 Hz);
13
C-NMR of 5h (CD
3
OD) ppm
(500 MHz): 39.51 (2 x CH
2
N), 40.48 (2 x CH
2
CO), 47.10, 47.27, 47.44, 47.61, 47.78,
47.90, 47.95, 48.07, 48.12, 48.24 (2 x 10 Ad-C), 55.07 (2 x OCH
3
), 101.37 (C-4),
110.27 (2 x C-2'), 115.04 (2 x C-5'), 120.79 (2 x C-6'), 122.77 (2 x C-1'), 127.07 (C-2
113
+ C-6), 129.77 (C-1 + C-7), 140.94 (2 x C-3'), 148.06 (2 x C-4'), 149.25 (C-3 + C-5),
183.23 (2 x CO); MS of 5h (Figure 14) m/z (% relative abundance): M
+
at 778
(absent), 719 (2.4), 705 (1.8), 698 (2.6), 692 (2), 646 (2.3), 616 (3.1), 611 (2.7), 581
(3), 575 (3.6), 562 (2.8), 530 (2.3), 463 (10.3), 442 (1.3), 430 (2.2), 422 (1.8), 392
(1.4), 340 (1.2), 338 (1.7), 318 (1.2), 305 (1.4), 289 (1), 281 (1.4), 273 (1.4), 251
(9.7), 237 (1.1), 195 (10.2), 194 (33.6), 180 (5), 164 (27), 151 (6.8), 135 (25.6), 121
(7), 119 (2.7), 116 (6.2), 107 (11.3), 106 (74.4), 94 (49.7), 93 (15.2), 91 (13.8), 81 (8),
79 (12.9), 77 (20.8), 67 (13.9), 65 (8.8), 57 (22), 55 (34.8), 53 (19), 46 (44.8), 45
(100), 44 (30.5), 43 (72), 41 (80.5).
IR of 5j (cm
-1
): 3435.7 (NH + OH intramolecularly H-bonded), 1790.6 (C=O,
ester), 1658.9 (C=O, ketone), 1558 (C=N mixed with C=C aromatic), 1498.8 (C=C
aromatic), 1273.2, 1169.9 (
as
and
s
C-O-C);
1
H-NMR of 5j (CD
3
COCD
3
) ppm
(500 MHz): 3.612 (s, 6H, 2 x CH
3
), 3.908 (s, 6H, 2 x OCH
3
), 4.084 (t, dist, 4H, 2 x
CH
2
-N), 4.609 (t, dist, 4H, 2 x CH
3
-CO), 5.791 (s, br, 2H, H
a
), 6.701 (d, 2H, 2x H
b
, J
= 15.5 Hz), 6.898-7.676 (m, 6H, 2 x Ar-H), 7.744 (d, 2H, 2 x H
c
, J = 15.5 Hz), 8.317
(s, 2H, 2 x NH);
13
C-NMR of 5j (CD
3
COCD
3
) ppm (500 MHz): 45.35 (2 x CH
2
N),
47.25 (2 x CH
2
CO), 60.85 (2 x OCH
3
), 105.75 (C-4), 111.80 (2 x C-2'), 116.85 (2 x
C-5'), 120.0 (2 x C-6'), 123.75 (2 x C-1'), 128.82 (C-2 + C-6), 134.93 (C-1 + C-7),
142.00 (2 x C-3'), 147.85 (2 x C-4'), 148.95 (C-3 + C-5), 170.40 (2 x thiazole C-2),
180.85 (2 x thiazole C-5), 192.75 (2 x CO); MS of 5j (figure 15) m/z (% relative
abundance): M
+
+1 707 (0.7), 664 (0.9), 658 (1.1), 618 (0.6), 561 (0.7), 496 (0.6), 491
(0.9), 387 (0.7), 307 (2.5), 293 (3), 268 (2), 242 (11), 171 (4), 167 (7), 150 (5), 149
(35), 142 (10), 115 (5), 100 (5.5), 99 (4.1), 83 (6.4), 73 (17.5), 63 (12), 55 (72.4), 45
(18.1), 43 (100).
114
1
H-NMR of 6a (CDCl
3
) ppm (300 MHz): 1.2516-2.3596 ((hump, br, 30H, 2 x Ad-
H), 1.4805 (t, 6H, 2 x OCH
2
CH
3
, J = 6.87 Hz), 2.3504 (s, 4H, 2 x CH
2
CO), 2.6306 (s,
2H, 2 x NH), 4.1516 (q, 4H, 2 x OCH
2
CH
3
, J = 6.87 Hz), 5.7825 (s, 1H, H
a
), 6.4541
(d, 2H, 2 x H
b
, J = 15.66 Hz), 6.9307 (d, 2H, 2 x H
A
, J
AB
= 8.25 Hz, ortho coupling),
7.0315 (s, 2H, 2 x H
X
), 7.10655 (d, 2H, 2 x H
B
, J = 8.25 Hz, ortho coupling), 7.5722
(d, 2H, 2 x H
c
, J = 15.66 Hz), 9.8134 (s, 1H, enol OH).
Table 4: Physicochemical Data of the Synthesized Compounds 5a-n and 6a-n.
OR
1
O
R
1
O
O O
n(H
2
C)
O O
(CH
2
)n
R
3
R
2
N
NR
2
R
3
(5a-n, 6a-n)
Cp
d
No
R
1
n
R
2
R
3
Reactio
n
Time
h
Yiel
d
%
m.p.
C
M. Formula
M.
wt
5a
-CH
3
1
NH
12
86
110
-15
C
45
H
54
N
2
O
8
75
0
5b
-CH
3
1
N
S
NH
H
3
CO
12
59
160
-
63
C
41
H
36
N
4
O
10
S
2
80
8
5c
-CH
3
1
N N
S
NH
H
3
C
24
63
170
-73
C
31
H
30
N
6
O
8
S
2
67
8
5d
-CH
3
1
HN
Cl
6
83
87-
93
C
29
H
32
Cl
2
N
2
O
8
60
7
115
5e -CH
3
1
N
Cl
Cl
4-5
95 85-
90
C
33
H
38
Cl
4
N
2
O
8
73
2
5f
-CH
3
1
NCH
2
CH
2
NH
H
3
C
H
3
C
1
69
160
-65
C
33
H
44
N
4
O
8
62
4
5g
-CH
3
1
NCH
2
CH
2
NH
C
2
H
5
C
2
H
5
1
62
150
-55
C
37
H
52
N
4
O
8
68
0
5h
-CH
3
2
NH
12
69
140
C
47
H
58
N
2
O
8
77
8
5i
-CH
3
2
N
S
NH
H
3
CO
36
47
170
-74
C
43
H
40
N
4
O
10
S
2
83
6
5j
-CH
3
2
N N
S
NH
H
3
C
36
43
120
-23
C
33
H
34
N
6
O
8
S
2
70
6
5k
-CH
3
2
HN
Cl
36
71
90-
95
C
31
H
36
Cl
2
N
2
O
8
63
5
5l
-CH
3
2
N
Cl
Cl
36
74
87-
93
C
35
H
42
Cl
4
N
2
O
8
76
0
5m
-CH
3
2
NCH
2
CH
2
NH
H
3
C
H
3
C
3-4
66
160
-65
C
35
H
48
N
4
O
8
65
2
5n
-CH
3
2
NCH
2
CH
2
NH
C
2
H
5
C
2
H
5
3-4
69
155
-60
C
39
H
56
N
4
O
8
70
8
116
6a
CH
2
CH
3
1
NH
4-5
71.5
115
-20
C
47
H
58
N
2
O
8
77
8
6b
CH
2
CH
3
1
N
S
NH
H
3
CO
12
66
137
-40
C
43
H
40
N
4
O
10
S
2
83
6
6c
CH
2
CH
3
1
N N
S
NH
H
3
C
12
72
105
-10
C
33
H
34
N
6
O
8
S
2
70
6
6d
CH
2
CH
3
1
HN
Cl
9
56
107
-10
C
31
H
36
Cl
2
N
2
O
8
63
5
6e
CH
2
CH
3
1
N
Cl
Cl
9
65
95-
100
C
35
H
42
Cl
4
N
2
O
8
76
0
6f
CH
2
CH
3
1
NCH
2
CH
2
NH
H
3
C
H
3
C
6
67
115
-20
C
35
H
48
N
4
O
8
65
2
6g
CH
2
CH
3
1
NCH
2
CH
2
NH
C
2
H
5
C
2
H
5
6
69
105
-
110
C
39
H
56
N
4
O
8
70
8
6h
CH
2
CH
3
2
NH
6
67
85-
90
C
49
H
62
N
2
O
8
80
6
6i
CH
2
CH
3
2
N
S
NH
H
3
CO
16
58
165
-70
C
45
H
44
N
4
O
10
S
2
86
4
6j
CH
2
CH
3
2
N N
S
NH
H
3
C
36
79
105
-
110
C
35
H
38
N
6
O
8
S
2
73
4
117
6k
CH
2
CH
3
2
HN
Cl
12
91
115
-
120
C
33
H
40
Cl
2
N
2
O
8
66
3
6l
CH
2
CH
3
2
N
Cl
Cl
12
64
125
-
130
C
37
H
46
Cl
4
N
2
O
8
78
8
6m
CH
2
CH
3
2
NCH
2
CH
2
NH
H
3
C
H
3
C
3-4
89
155
-60
C
37
H
52
N
4
O
8
68
0
6n
CH
2
CH
3
2
NCH
2
CH
2
NH
C
2
H
5
C
2
H
5
3-4
81
155
-60
C
41
H
60
N
4
O
8
73
6
5.2.5. 1,7-Bis(4-(4-substituted sulfanilamido)acyloxy)-3-(methoxyphenyl)-1,6-
heptadiene-3,5-dione (7a,b) and 1,7-bis(4-(4-substituted sulfanilamido)-
acyloxy)-3-(ethoxyphenyl)-1,6-heptadiene-3,5-dione (7c-f)
O
OR
1
O
R
1
O
O O
(CH
2
)n
O O
(CH
2
)n
HN
NH
R
4
HNO
2
S
SO
2
NHR
4
(7a-f)
Following the same procedure as for the preparation of compounds 5a-n and 6a-n,
the appropriate sulfonamide derivative (0.01 mol) was added to a solution of di-O-
chloroacylcurcumin 3a,b or di-O-chloroacylethyl curcumin 3c,d (0.005 mol) in EtOH
(30 ml) containing triethylamine (5 drops). The mixture was heated under reflux for
the specified time (Table 5), EtOH was evaporated under reduced pressure and the
residue was purified by elution on a column of silica gel using a mixture of toluene-
EtOH (97:3 v/v) to give compounds 7a-f (Table 5).
118
Table 5: Physicochemical Data of the Synthesized Compounds (7a-f)
O
OR
1
O
R
1
O
O O
(CH
2
)n
O O
(CH
2
)n
HN
NH
R
4
HNO
2
S
SO
2
NHR
4
(7a-f)
Cpd
No
R
1
n R
4
Reaction
Time
h
Yield
%
m.p.
C
M. Formula M. wt
7a
CH
3
1
H
16
69
155-60
C
37
H
36
N
4
O
12
S
2
792
7b
CH
3
1
N
N
12
50
175-78
C
45
H
40
N
8
O
12
S
2
948
7c
C
2
H
5
1
H
18
63
143-48
C
39
H
40
N
4
O
12
S
2
820
7d
C
2
H
5
1
N
N
24
60
185-90
C
47
H
44
N
8
O
12
S
2
976
7e
C
2
H
5
2
H
18
78
115-20
C
41
H
44
N
4
O
12
S
2
848
7f
C
2
H
5
2
N
N
24
57
115-20
C
49
H
48
N
8
O
12
S
2
1004
5.2.6. Di-O-Adamantoylcurcumin (8a) and di-O-adamantoylethyl curcumin (8b)
-OCO
O O
OCO-
R
1
O OR
1
(8a) R
1
= CH
3
(8b) R
1
= C
2
H
5
119
Adamantoyl chloride (0.03 mol) dissolved in the least amount of EtOH (10 ml)
was added to a mixture of curcumin (1) or ethyl curcumin (2) (0.01 mol) and Na
2
CO
3
(2 g) in acetone (50 ml). The mixture was heated under reflux for 9-19 hrs, filtered
and evaporated to dryness. The residue was extracted with EtOAc (3 x 25 ml). The
combined organic layer was dried over anhydrous MgSO
4
(5 g), filtered, evaporated
and crystallized from EtOH (15 ml) to give a yellow precipitate of 8a,b ( Table 6).
IR of 8a (cm
-1
): 3435.8 (OH intramolecularly H-bonded), 1750.4 (C=O, ester),
1630.9 (C=O, ketone), 1599, 1507.4 (C=C aromatic), 1255.2, 1122.8 (
as
and
s
C-O-
C);
1
H-NMR of 8a (CDCl
3
) ppm (300 MHz): 1.2342-2.1929 (hump, br, 30H, 2 x
Ad-H), 3.8467 (s, 6H, 2 x OCH
3
), 5.8475 (s, 2H, H
a
), 6.5539 (d, 2H, 2 x H
b
, J = 15.9
Hz), 7.0076 (d, 2H, 2 x H
A
, J
AB
= 8.25 Hz, ortho coupling), 7.0974 (s, 2H, 2 x H
X
),
7.1340, 7.1608 (dd, 2H, 2 x H
B
, J = 1.65 Hz and 8.04 Hz, meta and ortho coupling),
7.6111 (d, 2H, 2 x H
c
, J = 15.9 Hz);
13
C-NMR of 8a (CDCl
3
) ppm (500 MHz):
56.01 (2 x OCH
3
), 27.88, 27.94, 36.48, 38.73, 38.81, 41.14, 76.79, 77.04, 77.29 (2 x
Ad-C), 101.75 (C-4), 111.48 (2 x C-2'), 121.16 (2 x C-5'), 123.32 (2 x C-6'), 124.03 (2
x C-1'), 129.18 (C-2 + C-6), 140.13 (C-1 + C-7), 141.95 (2 x C-3'), 151.52 (2 x C-4'),
175.60 (C-3 + C-5), 183.16 (2 x CO); MS of 8a (figure 16) m/z (% relative
abundance): M
+
692 (2.1), 673 (3.7), 657 (2.4), 648 (2.3), 632 (3.2), 626 (2.1), 618
(2.3), 567 (2), 532 (2.17), 518 (2.75), 513 (2), 509 (2.5), 502 (2.4), 497 (1.7), 475
(2.1), 474 (2.2), 459 (2.3), 453 (4.3), 437 (2), 429 (1.34), 408 (1.3), 394 (1.6), 386
(1.35), 364 (1.26), 359 (3), 329 (1.2), 305 (1), 278 (1.2), 264 (1.5), 262 (2), 220 (1.7),
196 (3.4), 180 (4.8), 177 (1.5), 163 (5.6), 152 (2), 135 (100), 123 (5), 119 (2.9), 107
(9.1), 105 (5), 95 (3.2), 93 (29), 91 (15), 81 (9.1), 79 (36), 77 (16.2), 69 (8.7), 65
(9.1), 57 (22), 55 (29), 45 (31.1), 44 (26.4), 43 (50), 41 (92).
120
1
H-NMR of 8b (CDCl
3
) ppm (300 MHz): 1.2516-2.3596 (hump, br, 30H, 2 x Ad-
H overlapping with t of OCH
2
CH
3
), 1.4805 (t, 6H, 2 x OCH
2
CH
3
, J = 6.87 Hz),
4.1611 (q, 4H, 2 x OCH
2
CH
3
, J = 6.97 Hz), 5.7825 (s, 1H, H
a
), 6.4541 (d, 2H, 2 x H
b
,
J = 15.66 Hz), 6.9307 (d, 2H, 2 x H
A
, J
AB
= 8.25 Hz, ortho coupling), 7.0315 (s, 2H, 2
x H
X
), 7.1065 (d, 2H, 2 x H
B
, J
AB
= 8.25 Hz, ortho coupling), 7.5722 (d, 2H, 2 x H
c
, J
= 15.66 Hz); 9.8134 (s, 1H, enol OH).
5.2.7. Di-O-Heptanoylcurcumin (8c) and di-O-heptanoylethyl curcumin (8d)
O
OR
1
O
R
1
O
O O
O O
(8c) R
1
= CH
3
(8d) R
1
= C
2
H
5
Heptanoyl chloride (0.03 mol) was added dropwise to an ice cold mixture of
curcumin (1) or ethyl curcumin (2) (0.015 mol) and Na
2
CO
3
(2 g) in acetone (50 ml).
The mixture was stirred at room temperature for 4-6 hrs, evaporated to dryness and
the residue was extracted with EtOAc (2 x 20 ml). The combined organic extracts
were dried over anhydrous MgSO
4
, evaporated and crystallized from EtOH to give a
yellow precipitate of 8c,d (Table 6).
IR of 8c (cm
-1
): 1768 (C=O, ester), 1629 (C=O, ketone), 1598, 1512 (C=C
aromatic), 1120, 1010 (
as
and
s
C-O-C);
1
H-NMR of 8c (CDCl
3
) ppm (300 MHz):
0.9068 (t, 6H, 2 x (CH
2
)
5
-CH
3
, J = 6.96 Hz), 1.3049-1.7861 (m, 16H, 2 x (CH
2
)
4
),
2.5849 (t, 4H, 2 x CH
2
CO, J = 7.32 Hz), 3.8672 (s, 6H, 2 x OCH
3
), 5.8543 (s, 2H,
H
a
), 6.5468 (d, 2H, 2 x H
b
, J = 15.75 Hz), 7.0475 (d, 2H, 2 x H
A
, J
AB
= 8.04 Hz, ortho
coupling), 7.11405, 7.146 (dd, 2H, 2 x H
B
, J = 1.8 Hz and 9 Hz, meta and ortho
coupling), 7.2563 (s, 2H, 2 x H
X
), 7.6184 (d, 2H, 2 x H
c
, J = 15.75 Hz).
121
IR of 8d (cm
-1
): 1769 (C=O, ester), 1629 (C=O, ketone), 1590, 1500 (C=C
aromatic), 1115, 1020 (
as
and
s
C-O-C);
1
H-NMR of 8d (CDCl
3
) ppm (300 MHz)
(figure 17): 0.90441 (t, 6H, 2 x (CH
2
)
5
-CH
3
, J = 6.96 Hz), 1.3269-1.5724 (m, 16H, 2
x (CH
2
)
4
), 1.7666 (t, 6H, 2 x OCH
2
CH
3
, J = 7.3 Hz), 2.5727 (t, 4H, 2 x CH
2
-CO, J =
7.3 Hz), 4.0865 (q, 4H, 2 x OCH
2
CH
3
, J = 6.96 Hz), 5.8384 (s, 2H, H
a
), 6.5400 (d,
2H, 2 x H
b
, J = 15.75 Hz), 7.0383 (d, 2H, 2 x H
A
, J
AB
= 8.4 Hz, ortho coupling),
7.1134 (d, 2H, 2 x H
B
, J
AB
= 7.5 Hz, ortho coupling), 7.2551 (s, 2H, 2 x H
X
), 7.6013
(d, 2H, 2 x H
c
, J = 15.75 Hz);
13
C-NMR of 8d (CDCl
3
) ppm (300 MHz) (figure 18):
14.164, 14.813, 22.592, 25.157, 28.874 (5 peaks for 5Cs of side chain), 31.569 (2 x
OCH
2
CH
3
), 34.157 (2 x -OCOCH
2
C
5
H
11
), 64.454 (2 x OCH
2
CH
3
), 101.874 (C-4),
112.477 (2 x C-2'), 121.034 (2 x C-5'), 123.316 (2 x C-6'), 124.133 (2 x C-1'),
133.812 (C-2 + C-6), 140.171 (C-1 + C-7), 141.728 (2 x C-3'), 150.911 (2 x C-4'),
171.743 (C-3 + C-5), 183.216 (2 x CO); Examination of 2D
1
H-NMR (COSY)
(CDCl
3
) (figure 19) and
1
H,
13
C-NMR (C,H correlation) (CDCl
3
) (figure 20)
provided further confirmation for the structure of compound 8d.
5.2.8. Di-O-(2-Thienoyl)curcumin (8e) and di-O-(2-thienoyl)ethyl curcumin (8f)
-OCO
O O
OCO-
R
1
O OR
1
S S
(8e) R
1
= CH
3
(8f) R
1
= C
2
H
5
Compounds 8e,f (Table 6) were prepared by adding thiophene-2-carbonyl
chloride (0.03 mol) adopting the previously mentioned procedure.
IR of 8e (cm
-1
): 3339.6 (OH intramolecularly H-bonded), 1746.7 (C=O, ester),
1672.9 (C=O, ketone), 1606.8, 1494.5 (C=C aromatic), 1278.3, 1122 (
as
and
s
C-O-
122
C);
1
H-NMR of 8e (DMSO) ppm (500 MHz) (figure 21): 3.861 (s, 6H, 2 x OCH
3
),
6.232 (s, 2H, H
a
), 7.051 (d, 2H, 2 x H
b
, J = 16 Hz), 7.32, 7.336 (dd, 2H, 2 x H
B
, J =
3.5 Hz and 7.8 Hz, meta and ortho coupling), 7.396 (d, 2H, 2 x H
A
, J
AB
= 7.5 Hz,
ortho coupling), 7.588 (s, 2H, 2 x H
X
), 7.697 (d, 2H, 2 x H
c
, J = 16 Hz), 7.856 (d,
2H, 2 x thienyl-5-H, J = 8 Hz), 8.037, 8.059 (dd, 2H, 2 x thienyl-4-H), J = 1 Hz and 4
Hz ortho and meta coupling), 8.112 (d, 2H, 2 x thienyl-3-H, J = 4 Hz).
IR of 8f (cm
-1
): 3431.1 (OH intramolecularly H-bonded), 1727 (C=O, ester),
1626.5 (C=O, ketone), 1596.7, 1508.7 (C=C aromatic), 1250.7, 1117.2 (
as
and
s
C-
O-C);
1
H-NMR of 8d (DMSO-d
6
) ppm (300 MHz): 1.2351 (t, 6H, 2 x 3 OCH
2
CH
3
,
J = 7.00 Hz), 4.1412 (q, 4H, 2 x OCH
2
CH
3
, J = 7.04 Hz), 6.2138 (s, 2H, H
a
), 7.0269
(d, 2H, 2 x H
b
, J = 15.93 Hz), 7.3222 (d, 2H, 2 x H
A
, J
AB
= 8.52 Hz, ortho coupling),
7.3364 (s, 2H, 2 x H
X
), 7.67525 (d, 2H, 2 x H
c
, J = 15.93 Hz), 7.3845 (d, dist, 2H, 2 x
H
B
, J
AB
= 8.52 Hz, ortho coupling), 7.5690 (d, dist, 2H, 2 x thienyl-H-4), 8.0264 (br,
2H, 2 x thienyl-H-5, J = 3.57 Hz), 8.1015 (d, 2H, 2 x thienyl-H-3, J = 3.57 Hz);
13
C-
NMR of 8f (DMSO) ppm (500 MHz): 40.13 (2 x CH
3
), 64.95 (2 X OCH
2
), 102.31
(C-4), 113.79 (2 x C-2'), 121.94 (2 x C-5'), 123.94 (2 x C-6'), 125.22 (2 x C-1'),
129.26 (C-2 + C-6), 131.84 (thienyl-C-4), 134.46 (thienyl-C-3), 135.72 (thienyl-C-5),
135.83 (thienyl-C-2), 140.30 (C-1 + C-7), 141.37 (2 x C-3'), 151.02 (2 x C-4'),
159.88 (C-3 + C-5), 183.69 (2 x CO); MS of 8f (figure 22) m/z (% relative
abundance): M
+
616 (0.7), 604 (1.6), 588 (1.3), 561 (0.8), 510 (1.3), 490 (0.6), 458
(0.9), 431 (0.9), 417 (2.6), 399 (0.9), 371 (0.5), 344 (1.2), 320 (1.6), 314 (3.5), 295
(0.9), 262 (0.6), 247 (1.3), 219 (1), 203 (2.3), 193 (1.7), 173 (1.7), 163 (3), 161 (3),
143 (57), 138 (11), 125 (8), 121 (6), 119 (7), 111 (13), 107 (9), 105 (8.5), 99 (4), 97
(3), 95 (13), 93 (11.5), 91 (11.5), 91 (13), 85 (29), 83 (26.6), 81 (8.5), 79 (10), 77
(6.3), 69 (15), 67 (16.2), 45 (18.4), 44 (68.6), 43 (100).
123
Table 6: Physicochemical Data of the Synthesized Compounds 8a-f
O
OR
1
O
R
1
O
O O
R
2
R
2
O O
(8a-f)
Cpd
No
R
1
R
2
Reaction
Time (h)
Yield
(%)
m.p. M. Formula M.wt
8a
CH
3
9
47
215
C
43
H
48
O
8
692
8b
C
2
H
5
16
36
220
C
45
H
52
O
8
720
8c
CH
3
C
6
H
13
4-5
63
99.5
C
35
H
44
O
8
606
8d
C
2
H
5
C
6
H
13
4-5
77
128-30
C
37
H
48
O
8
620
8e
CH
3
S
4
81.5
199-203
C
31
H
24
O
8
S
2
588
8f
C
2
H
5
S
6
51
217-19
C
33
H
28
O
8
S
2
616
124
5.3. Anticancer Screening
5.3.1. Materials and Methods
5.3.1.1. Cytotoxicity to Mammalian Cells
The cytotoxic activity of the tested compounds 1, 2, 3a, 3c, 3d, 5a-k, 6a-h, 6m,n,
7a, 8a, 8c-f was determined against human cancer cell line SK-MEL (malignant,
melanoma), KB (epidermal carcinoma, oral), BT-459 (ductal carcinoma, breast), and
SK-OV-3 (ovary carcinoma). Vero cells, derived from monkey kidney fibroblasts,
and LLC-PK1, from pig kidney epithelial tissue, were used representing noncancerous
cells. The assay was performed in 96-well tissue culture treated microplates according
to the neutral red staining procedure as modified by Borenfreund and Peurner
[193]
. The
results are reported in Table 7 and represented in Charts 8-11.
125
Table 7: Cytotoxicity of the Synthesized Compounds to Mammalian
Cells
IC 50 value (M)*
Compd No
CANCER CELLS
NONCANCER
CELLS
SK-MEL KB BT-549 SK-OV-3 VERO LLC-PK1
1 13.75 >25 10.5 NA NC NC
2 16 >25 18.0 NA NC NC
3a 12.5 >25 10.0 NA NC NC
3c NA NA NA NA NC NC
3d NA NA NA NA NC NC
5a 15.0 >25 19.4 NA NC NC
5b 15.0 23 18.1 NA NC 22.5
5c 7.5 22.5 8.75 NA 23.0 22.5
5d NA NA NA NA NC NC
5e 7.0 19.5 6.75 NA >25 20.0
5f 15.0 NA 20.0 22.5 NA NC
5g 8.5 24.0 16.3 15.5 19.5 NC
5h 18.5 >25 20.50 NA NC NC
5i 23.8 >25 23.75 NA NC NC
5j 7.5 16.3 4.25 NA 15.0 15.5
5k 15.0 11.5 11.00 23.8 12.5 16.8
6a NA NA NA NA NC NC
6b 4.75 NA NA 2.8 NC NC
6c 12.5 11.5 12.00 15.8 11.5 13.0
6d 13.3 12.5 9.00 NA 11.0 6.8
6e 14.5 21.5 11.0 13.5 15.0 13.8
6f NA NA 11.50 NA NC 22.0
6g 11.5 11.5 10.00 14.8 5.0 5.8
6h NA 15 NA NA NC NC
6m NA NA NA NA NC NC
6n NA NA NA NA NC NC
7a 18.8 14 10.5 22.5 11 15.75
8a NA NA NA NA NC NC
8c NA NA NA NA NC NC
8d NA NA NA NA NC NC
8e NA NA NA NA NC NC
8f NA NA NA NA NC NC
Doxorubicin
0.55 <0.55 <0.55 0.8 >5.0 0.75
*Stock solution = 5mM in DMSO; Test concentrations = 25, 8.33, 2.78 M
NA = not active up to 25 M.
NC = not cytotoxic up to 25 M
126
SK-MEL = Human malignant, Melanoma
KB = Human Epidermal Carcinoma, Oral
BT-549 = Ductal Carcinoma, Breast
SK-OV -3 = Human Ovary carcinoma
Vero = Monkey Kidney Fibroblasts
LLC-PK1 = Pig Kidney Epithelial
127
0
5
10
15
20
25
30
d
o
x
o
r
u
b
i
c
i
n 1 2
3
a
3
c
3
d
5
a
5
b
5
c
5
d
5
e
5
f
5
g
5
h 5
i
5
j
5
k
6
a
6
b
6
c
6
d
6
e
6
f
6
g
6
h
6
m
6
n 7
8
a
8
c
8
d
8
e
8
f
SK-MEL
Compound number
Chart 8: Cytotoxic Effect of the Synthesized Compounds to SK-Mel Cells
: Most Cytotoxic Compounds
: Less cytotoxic Compounds
: Inactive Compounds
IC
50
128
0
5
10
15
20
25
30
d
o
x
o
r
u
b
ic
in 1 2
3
a
3
c
3
d
5
a
5
b
5
c
5
d
5
e
5
f
5
g
5
h 5
i
5
j
5
k
6
a
6
b
6
c
6
d
6
e
6
f
6
g
6
h
6
m
6
n 7
8
a
8
c
8
d
8
e
8
f
KB
Compound number
Chart 9: Cytotoxic Effect of the Synthesized Compounds to KB Cells
: Most Cytotoxic Compounds
: Less cytotoxic Compounds
: Inactive Compounds
IC
50
129
0
5
10
15
20
25
30
d
o
x
o
r
u
b
ic
in 1 2
3
a
3
c
3
d
5
a
5
b
5
c
5
d
5
e
5
f
5
g
5
h 5
i
5
j
5
k
6
a
6
b
6
c
6
d
6
e
6
f
6
g
6
h
6
m
6
n 7
8
a
8
c
8
d
8
e
8
f
BT-549
Compound number
Chart 10: Cytotoxic Effect of the Synthesized Compounds to BT-549 Cells
: Most Cytotoxic Compounds
: Less cytotoxic Compounds
: Inactive Compounds
IC
50
130
0
5
10
15
20
25
30
d
o
x
o
r
u
b
ic
in 1 2
3
a
3
c
3
d
5
a
5
b
5
c
5
d
5
e
5
f
5
g
5
h
5
i
5
j
5
k
6
a
6
b
6
c
6
d
6
e
6
f
6
g
6
h
6
m
6
n 7
8
a
8
c
8
d
8
e
8
f
SK-OV-3
Compound number
Chart 11: Cytotoxic Effect of the Synthesized Compounds to SK-OV-3 Cells
: Most Cytotoxic Compounds
: Less cytotoxic Compounds
: Inactive Compounds
IC
50
131
5.3.1.2. Interaction with Topoisomerases
Measurement of the catalytic activity of topoisomerase I (topo I) was based on the
conversion of supercoiled DNA to relaxed DNA. Cleavage complex stabilization was
assayed by measuring the nicked DNA produced in the presence of the tested
compounds. Human topo I and the topo I drug kit were purchased from Topogen Inc.
(Columbus, Ohio). A supercoiled plasmid DNA (pHOT1) with a high affinity topo I
recognition element was used as substrate. Enzyme activity was assayed in a total
volume of 20 l containing 250 ng of DNA, test compound, 2 to 4 U of purified
enzyme, 10 mM EDTA, 0.15 mM NaCl, 0.1% bovine serum albumin (BSA), 0.1 mM
spermidine, and 5% glycerol. The reaction mixture was incubated at 37
o
C for 30 min.
Reactions were then terminated by the addition of 1% sodium dodecyl sulfate (SDS)
followed by treatment with proteinase K (50 g/ml) at 37
o
C for 30 min. DNA was
extracted with chloroform-isoamyl alcohol (24:1 v/v) and analyzed by electrophoresis
on a 1% agarose gel in TAE buffer (40 mM Tris acetate, 2 mM EDTA, pH 8.5). The
gel was stained with ethidium bromide, destained in water, and photographed on a
UV transilluminator followed by a densitometric analysis using NIH image software
1.52. Enzyme activity was measured as a percentage of substrate DNA converted to
product. The concentration of test compound that prevented 50% of the substrate
from being converted into product (IC
50
) was calculated.
For the determination of cleavage complex formation activity with topo I, the
assay was performed with a minimum of 4U of purified enzyme as described above,
except that ethidium bromide was included in both the agarose gel and running buffer
to resolve nicked DNA from supercoiled or relaxed species. Drug-induced
stabilization of the cleaved complex was determined in terms of the percent nicked
DNA produced.
132
The relaxation activity of Topo II was analyzed in the same manner, except that
the reaction mixture contained 10 mM Tris-HCl (pH 7.9), 50 mM NaCl, 50 mM KCl,
5 mM MgCl
2
, 0.1 mM EDTA, 15 g/mL bovine serum albumin (BSA), 1 mM ATP
and 4 units Topo IIa. Decatenation of kDNA (TopoGEN Inc.) was performed in the
same condition except that supercoiled plasmid DNA was substituted by 400 ng
kDNA.
All compounds tested 1, 2, 3a, 3c, 3d, 5a-k, 6a-e, 6m,n, 7a, 8a, 8c-f were
dissolved in DMSO provided that the DMSO concentration did not exceed 2.5% in all
the assays and a DMSO control was always included. The results are reported in
Table 8.
133
Table 8: Anticancer Activity of the Synthesized Compound (Inhibition
of Topoisomerase Activity)
IC 50 value (M)
Sample
Topoisomerase I Topoisomerase II
cleavage* catalytic** cleavage* catalytic**
1 NA NA ND 15
2 NA NA ND 7.5
3a NA NA ND 20
3c NA NA ND 12
3d NA NA ND 20
5a NA NA ND 4
5b NA NA ND 15
5c NA NA ND 15
5d NA NA ND 0.35
5e NA NA ND 5
5f NA NA ND 16
5g NA NA ND 3
5h NA NA ND 18
5i NA NA ND 19
5j NA NA ND 3
5k NA NA ND 2.9
6a NA NA ND 3.4
6b NA NA ND 12
6c NA NA ND 2.7
6d NA NA ND 4.1
6e NA NA ND 3.4
6f NA NA ND 4
6g NA NA ND 10
6h NA NA ND 2.1
6m NA NA ND NA
6n NA NA ND 7.1
7a NA NA ND 3
8a NA NA ND 4.5
8c NA NA ND 2.9
8d NA NA ND 3
8e NA NA ND 8
8f NA NA ND 23
ND = not determined NA = not active up to 25 M.
*Cleavage complex stabilization activity (similar to camptothecin for topo I and
etoposide for topo II).
**Inhibition of catalytic activity of topoisomerase i.e. relaxation of
supercoiled DNA by topoisomerase I and decatenation of KDNA by
topoisomerase II.
134
8. References
1. Anderson, A.M.; Mitchell, M.S. and Mohan, R.S.; J. Chem. Ed.(2000),77: 359-
360.
2. Lampe,V; Chem. Ber. (1918), 51: 1347.
3. Ammon, H.P and Wahl, M.A; Planta Med. (1991), 57: 1-7.
4. Kunchandy, E. and Rao, M.N.A.; Int. J. Pharm. (1990), 58, 237-240.
5. Soudamini, K.K. and Kuttan R.; J. Ethnopharmacol. (1989), 27: 227-233.
6. Toda, S.; Miyase, T.; Arichi, H.; Tanizawa, H. and Takino, Y.; Chem.Pharm. Bull.
(1985), 33: 1725-1728.
7. Jovanovic, S.V.; Boone, C.W.; Steenken, S.; Trinoga, M. and Kaskey, R.B.; J. Am.
Chem. Soc. (2001),123: 3064-3068.
8. Mukhopadhyay, A.; Basu, N.; Ghatak, N. and Gujral P.K.; Agents Actions (1982),
12: 508-115.
9. Satoskar, R.R.; Shah, S.J. and Shonoy, S.G; Int. J. Clin. Pharm. Ther. Toxicol.
(1986), 24: 651-654.
10. Sharma, O.P.; Biochem. Pharmacol. (1976), 25: 1811-1812.
11. Srimal, R. and Dhawan, B.N., J. Pharm. Pharmacol. (1973), 25: 447-452.
12. Rao, T.S.; Basu, N. and Siddiqui, H.H.; Indian J. Med. Res. (1982), 75: 574-578.
13. Mehta, K.; Pantazis, P.; McQueen, T. and Aggarwal, B.B.; Anticance Drugs
(1997),
8: 470-481.
14. Huang, M.T.; Smart, R.C.; Wong, C.Q. and Conney, A.H.; Cancer Res.(1988),
48:
5941-5946.
15. Huang, M.T.; Newmark, H.L. and Frenkel, K.; J. Cell Biochem. (1997), 27: 26-34.
16. Huang, M.T.; Lou, Y.R.; Ma, W.; Newmark, H.L.; Reuhl, K.R. and Conney,
A.H.;
Cancer Res. (1994), 54: 5841-5847.
17. Kelloff, G.J.; Boone, C.W.; Crowell, J.A.; Steele, V.E.; Lubet, R. and Sigman,
C.C.;
Biomarkers Prev. (1994), 3: 85-98.
18. Kumar, S; Narian, U.; Tripathi, S. and Misra, K.; Bioconjugate Chem.(2001), 12:
135
464-469.
19. Mazumder, A.; Raghavan, K.; Weinstein, J.; Kohn, K.W. and Pommier, Y.;
Biochem. Pharmacol. (1995), 49: 1165-1170.
20. Lin, L.I.; Ke, Y.F.; Ko, Y.C. and Lin, J.K.; Oncology (1998), 55: 349-353.
21. Kiso, Y.; Suzuki, Y. and Watanabe, N. et al; Planta Med. (1983), 49: 185-187.
22. Rajakrishnan, V.; Viswanathan, P.; Rajasekharan, K.N. and Menon, V.P.;
Phytother. Res. (1999), 13: 571-574.
23. Masuda, T.; Maekawa, T.; Hidaka, K.; Bando, H; Takeda, Y. and Yamaguchi, H.;
J.
Agric. Food Chem. (2001), 49: 2539-2547.
24. Ruby, A.J.; Kuttan, G. and Babu K.D.; Cancer Lett. (1995), 94: 79-83.
25. Sreejayan, R.M.N.; J. Pharm. Pharmacol. (1994), 46: 1013-1016.
26. Kelloff, G.J.; Crowell, J.A.; Hawk, E.T.; Steele, V.E.; Lubet, R.A.; Boone, C.W.;
Covey, J.M.; Doody, L.A.; Omenn, G.S.; Greenwald, P.; Hong, W.K.; Parkinson,
D.R.; Bagheri, D.; Baxter, G.T.; Blunden, M.; Doelts, M.K.; Eisenhauer, K.M.;
Johnson, K.; Knapp, G.G.; Longfellow, D.G.; Malone, W.F.; Nayfield, S.G.;
Sefried, H.E.; Swall, L.M. and Sigman C.C.; J. Cell Biochem. (1996), 26S: 54-71.
27. Noguchi, N; Komuro, E.; Niki, E. and Willson, R.L.; J. Jpn. Oil Chem.Soc.
(1994),
43: 1045-1051.
28. Khopde, S.M.; Priyadarsini, K.I.; Venkatesan, P. and Rao, M.N.A.; Biophys.
Chem.
(1999), 80: 85-91.
29. Jovanoic, S.V.; Steenken, S.; Boone, C.V. and Simic, G.; J. Am. Chem. Soc.
(1999),
121: 9677-9681.
30. Anto, R.J.; Kuttan, G.; Babu, K.V.D. and Rajasekharan, K.N.; Int. J.Pharmaceut.
(1996), 131: 1.
31. Gorman, A.A.; Hamblett, I.; Srinavasan, V.S. and Wood, P.D.; Photochem.
Photobiol. (1994), 59: 389.
32. Priyadarsini, K.I.; Free Radicals Biol. Med. (1997), 23: 1997.
33. Priyadarsini, K.I.; Devasagayam, T.P.A.; Rao, M.N.A. and Guha, S.N.; Radiat.
Phys. Chem. (1999), 54: 551.
136
34. Frankel, E.N.; Lipid Oxidation; The Oily Press: Dundee, Scotland, (1998):129-
160.
35. Masuda, T; Hidaka, K.; Shinohara, A.; Maekawa, T.; Takeda, Y. and Yamaguchi,
H.; J. Agric. Food Chem. (1999), 47: 71-77.
36. Sugiyama, Y.; Kawakishi, S. and Osawa, T.; Biochem. Pharmacol. (1996), 52:
519.
37. Flynn, D.L.; Rafferty, M.F. and Boctor, A.M.; Leukotrienes Med. (1996),22: 357-
360.
38. Huang, M.T.; Lysz, T.; Ferrado, T.; Abidi, T.F.; Laskin, J.D. and Conney, A.H.;
Cancer Res. (1991), 51: 813-819.
39. Ammon, H.P.T.; Anazodo, M.I.; Safayhi, H.; Dhawan, B.N. and Srimal, R.;
Planta
Medica (1992), 58: 226.
40. Kiuchi, F.; Iwakami, S.; Shibuya, M.; Hanaoka, F. and Sankawa, U.; Chem.
Pharm.
Bull. (1992), 40: 387-391.
41. Cipriani, B.; Borsellino, G.; Knowles, H.; Tramonti, D.; Cavaliere, F.; Bernardi,
G.;
Battistini, L. and Brosnan, C.F.; J. Immunol. (2001), 167: 3454-3462.
42. Rajakrishnan, V. and Menon, V.P.; Cell Biol. Toxicol. (2001), 17: 11-22.
43. Joe, B. and Lokesh, B.R.; Mol. Cell Biochem.(2000), 203: 153-161.
44. Li, M.S.; Rafiee, P; Johnson, C.P.; Lamirand, T.H.; Fisher, P.J.; Olds, C.; Roza,
A.M.; Adams, M.B. and Binion, D.G.; Surg. Forum (2000), 51: 68-70.
45. Pan, M.H.; Lin-Shiau, S.Y. and Lin, J.K.; Biochem. Pharmacol. (2000), 60:
1665-
1676.
46. Graham, J.G.; Quinn, M.L.; Fabricant, D.S. and Farnsworth, N.R.; J.
Ethanopharmacol. (2000), 73: 347-377.
47. Pal, S.; Choudhuri, T.; Chattopadhyay, S.; Bhattacharya, A.; Datta, G.K.; Das, T.
and Sa, G. Biochem. Biophys. Res. Commun. (2001), 288: 658-655.
48. Azuine, M.A. and Bhide, S.V.; Nutr. Cancer (1992), 17: 77-83.
49. Huang, M.T.; Wang, Z.Y.; Georgiadis, C.A.; Laskin, J.D. and Conney, A.H.;
Carcinogenesis (1992), 13: 2183-2186.
50. Nagabhushan, M. and Bhide, S.V.; J. Am. Coll. Nutr. (1992), 11: 192-198.
137
51. Matthes, H.W.D.; Luu, B. and Ourisson, G.; Phytochemistry (1980), 19: 2643.
52. Syu, W.J.; Shen, C.C.; Don, M.J.; Ou, J.C.; Lee, G.H. and Sun, C.M.; J.Nat. Prod.
(1998), 61: 1531-1534.
53. Kuo, M.L.; Huang, T.S. and Lin, J.K.; Biochem. Biophys. Acta (1996),1317: 95.
54. Nogaki, A; Satoh, K.; Iwasaka, K.; Takano, H.; Takahama, M.; Ida, Y. and
Sakagami, H.; Anticancer Res. (1998), 18: 3487-3491.
55. Rao, C.V.; Rivenson, A.; Simi, B. and Reddy, B.S.; Cancer Res. (1995), 55: 259.
56. Huang, M.T.; Ma, W.; Lu, Y.P.; Chang, R.L.; Fisher, C.; Manchand, P.S.;
Newmark, H.L. and Conney, A.H.; Carcinogenesis (1995), 16: 2493.
57. Conney, A.H.; Lysz, T.; Ferraro, T.; Abidi, T.F.; Manchand, P.S.; Laskin, J.D. and
Huang, M.T.; Adv. Enzyme Regul. (1991), 31: 385.
58. Sindhwani,P.; Hampton, J.A.; Baig, M.M.; Keck, R.W. and Selman, S.H.; Surg.
Forum (2000), 51: 625-626.
59. Mukhopadhyay, A.; Bueso-Ramos, C.; Chatterjee, D.; Pantazis, P. and Aggarwal,
B.B.; Oncogene (2001), 20: 7597-7609.
60. Dorai, T.; Cao, Y.C.; Dorai, B.; Buttyan, R. and Katz, A.E.; Prostate (NY.,
U.S.)(2001), 47:293-303.
61. Pal, S.; Choudhuri, T.; Chattopadhyay, S.; Bhattacharya, A.; Datta, G.K.; Das, T.
and Sa, G.; Biochem. Biophys. Res. Commu. (2001), 288: 658-665.
62. Arbiser, J.L.; Klauber, N.; Rohan, R.; Van-Leeuwen, R.; Huang, M.T.; Fisher, C.;
Flynn, E. and Byers, H.R.; Mol. Med. (1998), 4: 376.
63. Mohan, R.; Sivak, J.; Ashton, P.; Russo, L.A.; Pham, B.Q.; Kasahara, N.;
Raizman,
M.B. and Fini, M.E.; J. Biol. Chem. (2000), 275: 10405.
64. Thaloor, D.; Singh, A.K.; Sidhu, G.S.; Prasad, P.V.; Kleinman, H.K. and
Maheshwari, R.K.; Cell Growth Differ. (1998), 9: 305.
65. Ishida, J.; Ohtsu, H.; Tachibana, Y.; Nakanishi, Y.; Bastow, K.F.; Nagai, M.;
Wang,
H.K.; Itokawa, H. and Lee, K.H.; Biorg. Med. Chem. (2002), 10: 3481-3487.
66. Simon, A.; Allais, D.P.; Duroux, J.L.; Basly, J.P.; Durand-Fontanier, S. and
Delage,
C.; Cancer Lett. (1998), 129: 111-116.
67. Mukundan, M.A.; Chacko, M.; Annapurna, V.V. and Krishnaswamy, K.;
Carcinogenesis (1993), 14: 493-496.
138
68. Kawamori, T.; Lubet, R.; Steele, V.E.; Kelloff, G.J.; Kaskey, R.B.; Rao, C.V. and
Reddy, B.S.; Cancer Res. (1999), 59: 597-601.
69. Bae, J.H.; Park, J.W. and Kwon, T.K.; Biochem. Biophys. Res. Commu. (2003),
303: 1073-1079.
70. Chen, Y.K.; Duniec, Z.M.; Liu, B.; Hagmann, W.; Gao, X.; Shimoji, K.I.;
Marnett,
L.J.; Johnson, C.R. and Honn, K.V.; Cancer Res. (1994), 54: 1574-1579.
71. Honn, K.V. and Dunn, J.R.; FEBS Lett. (1982), 139: 65-68.
72. Werner, E.J.; Waltenga, R.W.; Dubowy, R.L.; Boone, S. and Suart, M.J.; Cancer
Res. (1985), 45: 561-563.
73. Najid, A.; Beneytout, J.L. and Tixier, M.; Cancer Lett. (1989), 46: 137-141.
74. Lin, J.K.; Pan, M.H. and Lin-Shiau, S.Y.; Biofactors (2000), 13: 153-158.
75. Shim, J.S.; Kim, D.H.; Jung, H.J.; Kim, J.H.; Lim, D.; Lee, S.K.; Kim, K.W.;
Ahn,
J.W.; Yoo, J.S.; Rho, J.R.; Shin, J. and Kwon, H.J.; Bioorg. Med. Chem. (2002),
10:
2439-2444.
76. Sharma, R.A.; Ireson, C.R.; Verschoyle, R.D.; Hill, K.A.; Williams, M.L.;
Leuratti,
C.; Manson, M.M.; Marnett, L.J.; Steward, W.P. and Gescher, A.; Clin. Cancer
Res.
(2001), 7:1452-1458.
77. Xu, K. and Thornalley, P.J.; Biochem. Pharmacol. (2001), 61:165-177.
78. Onoda, M. and Inano, H.; Nitric Oxide (2000), 4: 505-515.
79. Churchill, M.; Chadburn, A.; Bilinski, R.T. and Bertagnolli, M.M.; J. Surg. Res.
(2000), 89: 169-175.
80. Hadi, S.M.; Asad, S.F.; Singh, S.; Ahmad, A.; IUBMB Life (2000), 50: 167-171.
81. Venkatesan, N.; Br. J. Pharmacol. (1998), 124: 425.
82. Rajakrishnan, V.; Viswanathan, P.; Rajasekaran, K.N.; Gunashekaran, G. and
Menon, V.P.; Med. Sci. Res. (1998), 26:715.
83. Thapliyal, R.; Deshpande, S.S. and Maru, G.B.; J. Environ. Pathol. Toxicol.
Oncol.
(2001), 20: 59-63.
139
84. Inano, H.; Onoda, M.; Inafuka, N.; Kubota, M.; Kamada, Y.; Osawa, T.;
Kobayashi,
H. and Wakabayashi, K.; Carcinogenesis (2000), 21: 1835-1841.
85. Ahmad, N.; Katiyar, S.K. and Mukhtar, H.; Curr. Probl. Dermatol. (2001), 29:
128-
139.
86. Donatus, I.A.; Argo, S. and Vermeulen, N.P.E.; Biochem. Pharmacol. (1990), 39:
1869-1875.
87. Soni, K.B.; Lahiri, M.; Chackradeo, P.; Bhide, S.V. and Kuttan, R.; Cancer Lett.
(1997), 115: 129-133.
88. Negi, P.S.; Jayaprakasha, G.K.; Rao, L.J.M. and Sakariah, K.K.; J. Agric. Food
Chem. (1999), 47: 4297-4300.
89. Jayaprakasha, G.K.; Negi, P.S.; Anandharamkrishnan, C. and Sakariah, K.K.; J.
Biosci. (2001), 56: 40-44.
90. Tsao, S.M. and Yin, M.C.; Yaowu shipin fenxi (2000), 8: 208-212.
91. Nose, M.; Koide, T.; Ogihara, Y.; Yabu, Y. and Ohta, N.; Biol. Pharm. Bull.
(1998),
21: 643-645.
92. F. Kiuchi, S. Nishizawa, A. Uchitani, H. Ohshima, K. Kondo and Y. Tsuda;
Chem.
Pharm. Bull. (1992),40: 3234
93. F. Kiuchi, Y. Goto, N. Sugimoto, N. Akao and Y. Tsuda; Chem. Pharm. Bull. 41:
1640-1643 (1993).
94. Yano, S.; Terai, M.; Shimizu, K.L.; Horie, S.; Futagami, Y.; Tsuchiya, S.;
Ikegami,
F.; Sekine, T.; Yamamoto, Y.; Fujimori, H.; Takamoto, K.; Saito, K.; Ueno, K.
and
Watanabe, K.; Nat. Med. (2000), 54: 318-324.
95. Yano, S.; Terai, M.; Shimizu, K.L.; Futagami, Y.; Horie, S.; Tsuchiya, S.;
Ikegami,
F.; Sekine, T.; Takamoto, K.; Saito, K.; Ueno, K. and Watanabe, K.; Nat. Med.
(2000), 54: 325-329.
96. Arbiser, J.L.; PCT Int. Appl. WO (2001), A 14 Jan. 200 1000 201.
97. Anderson, G.T.; Ptchelintsev, D. and Traudt, M.; Eur. Patent Appl. (2001)
1108419
A1 200 10 620.
98. Vogel and Pelletier; J. Pharm. (1815), 2: 50.
140
99. Nurfina, A.N.; Reksohadiprodjo, M.S.; Timmerman, H.; Jeni, U.A.; Sugiyanto, D.
and van der Goot, H.; Eur. J. Med. Chem. (1997), 32: 321-328.
100. Daube, F.V.; Ber. (1870), 3: 609.
101. Lampe, V.; Milobedzka, J. and Kostanecki, St. v.; Ber. (1910), 43: 2163.
102. Lampe, V. and Milobedzka, J.; Ber. (1913), 46: 2235.
103. Srinivasan, K.R.; J. Pharm. Pharmacol. (1953), 5: 448.
104. Suga, T.; Asakawa, Y. and Iwata, N.; Chem. Ind. (1971), 766.
105. Asakawa, Y.; Genjida, F; Hayashi, S. and Matsuura, T.; Tetrahedron Letters
(1969), 3235.
106. Asakawa, Y.; Bull. Chem. Soc. Jpn. (1970), 43: 575, 2223.
107. Roughley, P.J. and Whiting, D.A.; J. Chem. Soc. Trans I (1973), 2379-2388.
108. Huang, X.S.; Chinese Condiment (1997), 8: 2-4.
109. Ma, J.; Jin, X.; Yang, L. and Liu, Z.L.; Phytochem. (2004), 65: 1137-1143.
110. Morikawa, T.; Tao, J.; Ueda, K.; Matsuda, H. and Yoshikawa, M.; Chem.
Pharm.
Bull. (2003), 51: 62-67.
111. Inoue, T.; Yakugaku Zasshi (1993), 113: 181-197.
112. Nagumo, S.; Kaji, N.; Inoue, T. and Nagai, M.; Chem. Pharm. Bull. (1993), 41:
1225-1257.
113. Shiratori, S.; Nagumo, S.; Inoue, T. Nagai, M. and Chi, H.J.; Chem. Pharm. Bull.
(1994), 42: 960-962.
114. Nagumo, S.; Ishizawa, S.; Nagai, M. and Inoue, T.; Chem. Pharm. Bull. (1996),
44: 1086-1089.
115. Li, G.; Xu, M.L.; Choi, H.G.; Lee, S.H.; Jahng, Y.D.; Lee, C.S.; Moon, D.C.;
Woo,
M.H. and Son, J.K.; Chem. Pharm. Bull. (2003), 51: 262-264.
116. Joe, Y.K.; Son, J.K.; Park, S.H.; Lee, I.J. and Moon, D.C.; J. Nat. Prod. (1996),
59: 159-160.
117. Son, J.K.; Arch. Pharm. Res. (1995), 18: 203-205.
141
118. Kim, S.H.; Lee, K. S.; Son, J.K.; Je, G. H.; Lee, J.S.; Lee, C.H. and Cheong,
C.J.;
J. Nat. prod. (1998), 61: 643-645.
119. Lee, S.W.; Lee, K.S. and Son, J.K.; Planta Med. (2000), 66: 184-186.
120. Lee, K.S.; Li, G.; Lee, C.S.; Woo, M.H.; Lee, S.H.; Jhang, Y.D. and Son, J.K.; J.
Nat. Prod. (2002), 65: 1707-1708.
121. Uehara, S.I.; Yasuda, I.; Akiyama, K.; Morita, H.; Takeya, K. and Itokawa, H.;
Chem. Pharm. Bull. (1987), 35: 3298-3304.
122. Kuroyanagi, M. and Natori, S.; Yakugaku Zasshi (1970), 90: 1467.
123. Flynn, D.L.; Belliotti, T.; Boctor, A.M.; Conner, D.T.; Kostlan, C.R.; Nies, D.E.;
Ortwine, D.F.; Schrier, D.J. and Sircar, J.C.; J. Med.Chem. (1991), 34: 518-525.
124. Ciamician, G. and Silber, P.; Ber. (1897), 30: 192.
125. Jackson, C.L. and Clarke, L., Amer. Chem. J. (1908), 39: 696; (1911), 45: 48.
126. Mazumder, A.; Neamati, N.; Sunder, S.; Schulz, J.; Pertz, H.; Eich, E. and
Pommier, Y.; J. Med. Chem. (1997), 40: 3057-3063.
127. Ishigami, Y., Suzuki, S. and Takizawa, Y.; J. Jpn. Soc. Colour Material (1995),
68: 331-336.
128. Hergenhahn, M.; Bertram, B.; Wiessler, M.; Sorg, B.L.; Ger. Patent DE 2337
(2001).
129. Mohri, K.; Watanabe, Y.; Yoshida, Y.; Satoh, M. ; Isobe, K.; Sugimoto, N. and
Tsuda, Y.; Chem. Pharm. Bull. (2003), 51: 1268-1272.
130. Begley, M.J.; Campbell, R.V.M.; Crombie, L.; Tuck, B. and Whiting, D.A.; J.
Chem. Soc. (C) (1971), 3634.
131. Yasue, M.; J. Jpn. Wood Res. Soc. (1965), 11: 146.
132. Morikawa, T.; Matsuda, H.; Sakamoto, Y.; Ueda, K. and Yoshikawa, M.; Chem.
Pharm. Bull. (2002), 50: 1045-1049.
133. Morohara M.; Sakurai, N.; Inoue T.; Kawai, K. and Nagai, M.; Chem. Pharm.
Bull.
(1997), 45: 820-823.
134. Jiang, Z.; Tanaka, T.; Inutsuka, C. and Kouno, I.; Chem. Pharm. Bull.(2001), 49:
737-740.
135. Sundaryono, A.; Nourmamode, A.; Gardat, C.; Fritsch, A. and Castellan, A.; J.
Mol. Structure (2003), 649: 177-190.
142
136. Rojas, J.; Dominguez, J.N.; Charris, J.E.; Lobo, G.; Paya, M. and Ferrandiz,
M.L.;
Eur. J. Med. Chem. (2002), 37: 699-705.
137. Nogueira, M.A.; Magalhaes, E.G.; Magalhaes, A.F.; Biloti, D.N.; Laverde, A.,
Jr.;
Pessine, F.B.T.; Carvalho, J.E.; Kohn, L.K.; Antonio, M.A. and Marsaioli, A.J.;
Il
Farmaco (2003), 58: 1163-1169.
138. K.M. Youssef, M.A. El-Sherbiny, F.S. El-Shafie, H.A. Farag, O.A. Al-Deeb and
S.A. Awadalla; Arch. Pharm. Pharm. Med. Chem. (2004), 337: 42-54.
139. Ross, L.; Barclay, C. and Vinqvist, M.R.; Org. Lett. (2000),10: 404-418.
140. Roth, G.N.; Chandra, A. and Nair, M.G.; J. Nat. Prod. (1998), 61: 542-545.
141. Snyder, R.D. and Arnone, M.R.; Mutation Res. (2002), 503: 21-35.
142. Martin-Cordero, C.; Lopez-Lazaro, M.; Galvez, M. and Avuso, M.J.; J. Enzyme
Inhib. Med. Chem. (2003), 18: 505-509.
143. Pabon, H.J.J; Rec. Tr. Chim. Pays-Bas (1964), 83: 379-386.
144. (a) Nielson, A.T. and Houliban, W.J.; Org. React. (1968), 16: 1; (b) Claisen, L.
and Claparede, A.; Ber. (1881), 14: 2460; (c) Schmidt, J.G.; Ber.(1881), 14:
1459;
In: Smith M.B.; Organic Synthesis (1994), McGraw Hill, Inc., NY, London,
Torento; Chapter 9: p. 857.
145. Robinson, T.P.; Ehlers, T.; Hubbard IV, R.B.; Bai, X.; Arbiser, J.L.; Goldsmith,
D.J. and Bowen, J.P.; Bioorg. Med. Chem. Letters (2003),13: 115-117.
146. Smith M.B.; Organic Synthesis (1994), McGraw Hill, Inc., NY, London,
Toronto;
Chapter 9: p.886.
147. Rumpel, W; Austrian Pat. Appl. 180, 258 (1954).
148. Sardjiman, S.S.; Reksohadiprodjo, M.S.; Hakim, L.; van der Goot, H. and
Timmerman, H.; Eur. J. Med. Chem. (1997), 32: 625-630.
149. Pedersen, U.; Rasmussen, P.B. and Lawesson, S.O.; Liebigs Ann.Chem. (1985),
1557.
150. Breitmaier, E.; In: Structure Elucidation by NMR in Organic Chemistry: A
Practical Guide (1999), John Wiley & Sons, NY,Toronto; Chapter 2: pp. 24-26.
143
151. Dyke, S.F.; Flyod, A.J.; Sainsbury, M. and Theobald, R.S.; In: Organic
Spectroscopy: An Introduction (1985), 2nd Edition, Longman: London, NY;
Chapter 4: pp. 129-130.
152. Oruc, E.E.; Rollas, S.; Kandemirli, F.; Shvets, N. and Dimoglo, A.S.; J. Med.
Chem. (2004), 48: 6760-6767.
153. Crabbe, P.; Maldonado, L.A. and Sanchez, I; Tetrahedron (1971), 27: 711-725.
154. Lin, A.J and Hoch, J.M.; Arzneim.-Forsch. (1984), 34: 640-642.
155. Omar, A.M.M.E..; Ahmed, IC; AboulWafa, O.M.; Hassan, A.M.; Ismail, K.A.;
El-
Din, M.M.M. and Mansour, N.; Eur. J. Med. Chem.(1994) 29: 25-32.
156. Luo, Q.; Li, J.; Liu, Z.; Chen, L.; Li, J.; Qian, Z.; Shen, Q.; Li, Y.; Lushington,
G.
H.; Ye, Q. and Nan, F.; J. Med. Chem. (2003), 46: 2631-2640.
157. Sebille, S.; de Tullio, P.; Becker, B.; Antoine, M.H.; Boverie, S.; Pirotte, B. and
Lebrun, P.; J.Med.Chem. (2005), 48: 614-621.
158. Clayton, A.; J. Chem. Soc. Part I (1908), 2016.
159. Smith M.B. ; In: Organic Synthesis (1994), McGraw Hill, Inc., NY, London,
Toronto; Chapter 2: pp.150-155.
160. Aldrich Handbookof Fine Chemicals and Laboratory Equipment: 2003-2004.
161. Smith M.B. ; In: Organic Synthesis (1994), McGraw Hill, Inc., NY, London,
Toronto; Chapter 2: pp.130-147.
162. Lopez-Rodriguez, M.L.; Porras, E.; Morcillo, M.J.; Benhamu, B.; Soto, L.J.;
Lavandera, J.L.; Ramos, J.A.; Olivella, M.; Campillo, M. and Pardo, L.; J. Med.
Chem. (2003), 46: 5638-5650.
163. Omar, A. M. M. E. and AboulWafa, O.M.; J. Pharm. Sci. (1982), 71: 983-986.
164. Leonardi, A.; Barlocco, D.; Montesano, F.; Cignarella, G.; Motta, G.; Testa, R.;
Poggesi, E.; Seeber, M.; De Benedetti, P.G. and Fanelli, F.; J. Med. Chem.
(2004),
47: 1900-1918.
165. Baraldi, P.G.; Tabrizi, M.A.; Preti, D.; Bovero, A.; Romagnoli, R.; Fruttarolo, F.;
Abdel Zaid, N.; Moorman, A.R.; Varani, K.; Gessi, S.; Merighi, S. and Borea,
P.A.; J. Med. Chem. (2004), 47: 1434-1447.
166. Wang, S.; Folkes, A.; Chuckowree, I.; Cockcroft, X.; Sohal, S.; Miller, W.;
Milton,
144
J.; Wren, S.P.; Vicker, N.; Depledge, P.; Scott, J.; Smith, L.; Jones, H.; Mistry,
P.;
Faint, P.; Thompson, D. and Cocks, S.; J. Med. Chem. (2004), 47: 1329-1338.
167. Romeo, G.; Materia, L.; Manetti, F.; Cagnotto, A.; Mennini, T.; Nicoletti, F.;
Botta, M.; Russo, F. and Minneman, K.P.; J. Med. Chem. (2003), 46: 2877-2894.
168. Dardonville, C. and Brun, R.; J. Med. Chem. (2004), 47: 2296-2307.
169. Silverstein, R.M. and Webster, F.X.; In: Spectrometric Identification of Organic
Compounds (1998), 6
th
Edition, John Wiley & Sons, Inc. NY, Toronto; Chapter
4:
p. 210.
170. Li, T. and Liu, L.; Annual Rev. Pharmacol. Toxicol. (2001) 41: 5377.
171. Roninson, I.B.; Broude, E.V. and Chang, B.D.; Drug Resis. Upd. (2001) 4: 303
313.
172. Lock, R. and Stribinskiene, L.; Cancer Res. (1996) 56: 40064012.
173. Pindur, U. Curr. Med. Chem., (2001), 8: 465-581.
174. Bischoll, G. and Hoffmann, Curr. Med. Chem. (2002), 9: 321-348.
175. Arcamone, F., Penco, S., Orezzi, P., Nicollela, V. and Pirelli, A.; Nature (1964),
203: 1064.
176. Finlay, A.C., Hochstein, F.A., Sobin, B.A. and Murphay, F.X.; J. Am.Chem. Soc.
(1951), 73: 341-344
177. Pindur, U. and Fischer, G.; Curr. Med. Chem. (1996), 3: 379.
178. Bailly, C. In: Advances in DNA sequence-specific Agents (1998), 3: 97-115.
179. Holtzel, C., Marotto, A. and Pindur, U.; Eur. J. Med. Chem. (2003), 38: 189-197.
180. Staker, B.L.; Feese, M.D.; Cushman, M.; Pommier, Y. ; Zembower, D.; Stewart,
L.
and Burgin, A.B.; J. Med. Chem. (2005), 48: 2336-2345.
181. Wall, M.E.et al; J.Am.Chem.Soc. (1966) 88: 3888-3890.
182. Hsiang, Y.H., Hertzberg, R.; Hecht, S.; Liu, L.F.; J. Biol. Chem.(1985), 260:
14873-14878.
183. Bjornsti, M.A.; Beneditti, P., Viglianti, G.A.and Wang, J.C.; Cancer Res. (1989),
49: 6318-6323.
145
184. Pommier,Y.; Kohlhagen, G.; Kohn, K.W.; Leteurtre, F.; Wani, M.C. et al.; Proc.
Natl. Acad. Sci. USA (1995), 92: 8861-8865.
185. Nitiss, J.L. and Wang, J.C.; Proc. Natl. Acad. Sci. USA (1988), 85: 7501-7505.
186. Chen, A.Y. and Liu, L.F.; Rev. Pharmacol. Toxicol. (1994), 34: 191-218.
187. Tanizawa, A. Kohn, K.W.; Kohlhagen, G.; Leteurtre, F. and Pommier, Y.;
Biochemistry (1995), 34: 7200-7206.
188. Sordet, O.; Khan, Q. A.; Kohn, K.W. and Pommier, Y.; Curr. Med.Chem. Anti-
Cancer Agents (2003), 3: 271-290.
189. Burke, P.J. and Koch, T.H.; J. Med. Chem. (2004), 47: 1193-1206.
190. Foye, W.O. and Sengupta, S.K.: Cancer Chemotherapy. In: Principles of
Medicinal Chemistry.; Foye, W.O.; Lemke, T.L. and Williams, D.A. (eds).
(1995),
Lippincott Williams and Wilkins, Chapter 37, p 839.
191. Doroshow, D.A. In: Anthracyclines and Anthracenediones; Lippincott-Raven
Press, Philadelphia, PA (1996), pp 409-434.
192. Kato, S.; Burke, P.J.; Fenick, D.J.; Taatjes, D.J.; Bierbaum, V.M.; et al.
Chem.Res.
Toxicol. (2000), 13: 509-516.
193. Borenfreud, E., and Peurner, J.; Toxicol. Lett. (1985), 24:119-124.
146
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