Med Chem Res
0
0
0
(
1H, s, H-3), 8.34–7.99 (2H, m, NH ), 7.56–7.30 (4H, m,
2
113.80 (C-1 ), 111.89 (C-3 ), 111.73 (C-5 ); MS m/z
1
3
?
401.0393 [MH ].
ar–H); C NMR (DMSO) d/ppm: 166.07 (C=O-2), 156.25
(
(
(
[
C-4), 152.25 (C-2c), 151.36 (C-11), 140.92 (C-2), 138.09
C-3), 136.08 (C-3c), 130.56 (C-7), 125.13 (C-9), 124.49
C-8), 117.22 (C-6), 116.63 (C-10); MS m/z 245.0149
N-((3-chloro-2-fluorophenyl)carbamoyl)-4-oxo-4H-thieno[3,2-
c]chromene-2-carboxamide (10) This compound was syn-
thesized following the general procedure using 3-chloro-2-
?
MH ].
fluorophenylisocyanate (0.51 ml) to give 10 (111 mg,
1
1
4 %, m.p. = 267–269 °C). H NMR (DMSO-d ) (d/
6
General procedure for the preparation
of compounds 7–11
ppm): 11.73 (1H, s, NH), 10.79 (1H, s, NH), 8.82 (1H, s,
3
1
H-3), 8.17–7.22 (7H, m, ar–H); C NMR (DMSO) d/ppm:
0
1
65.97 (C=O-2), 161.10 (C-2 ), 155.49 (C-4), 151.74 (C-
Isocyanate derivatives (4.0 mmol) were added to a stirred
mixture of 6 (500 mg, 2.0 mmol) in toluene (20 ml). The
solution was then stirred overnight at 110 °C, evaporated
and purified by column chromatography (CH Cl :
2c), 150.12 (C-11), 142.01 (C-3), 141.17 (C-2), 131.23
0
(NH–C=O–NH), 131.06 (C-7), 127.78 (C-5 ), 126.09 (C-
0
4 ), 124.53 (C-3c), 123.01 (C-9), 122.87 (C-8), 121.61 (C-
0
0
2
2
6 ), 120.01 (C-3 ), 117.07 (C-6), 116.02 (C-10), 115.92 (C-
0
?
1 ); MS m/z 417.0403 [MH ].
MeOH = 30: 1) to give 7–11 as white solids.
N-((2-fluorophenyl)carbamoyl)-4-oxo-4H-thieno[3,2-c]chro-
mene-2-carboxamide (7) This compound was synthesized
following the general procedure using 2-difluorophenyliso-
N-((5-chloro-2-phenoxyphenyl)carbamoyl)-4-oxo-4H-thieno[3,2-
c]chromene-2-carboxamide (11) This compound was syn-
thesized following the general procedure using 5-chloro-2-
cyanate (0.46 ml) to give 7 (138 mg, 17 %, m.p. =
1
phenoxyphenylisocyanate (1000 mg) to give 11 (214 mg,
1
2
37–239 °C). H NMR (DMSO-d ) (d/ppm): 11.70 (1H, s,
11 %, m.p. = 279–281 °C). H NMR (DMSO-d ) (d/
6
6
NH), 10.75 (1H, s, NH), 8.83 (1H, s, H-3), 8.22–7.18 (8H, m,
ppm): 11.73 (1H, s, NH), 11.14 (1H, s, NH), 8.77 (1H, s,
13
13
ar–H); C NMR (DMSO) d/ppm: 166.37 (C=O-2), 162.30
H-3), 8.42–7.05 (12H, m, ar–H); C NMR (DMSO) d/
0
0
0
(
C-2 ), 156.09 (C-4), 152.14 (C-2c), 150.32 (C-11), 141.51
ppm: 165.06 (C=O-2), 158.51 (C-2 ), 158.40 (C-7 ), 157.93
0
0
(
C-3 ), 140.87 (C-2), 132.63 (NH–C=O–NH), 130.90 (C-7),
(C-6 ), 156.36 (C-4), 153.91 (C-2c), 152.19 (C-11), 141.85
0
0
0
1
1
1
29.01 (C-5 ), 128.79 (C-4 ), 125.67 (C-3c), 125.31 (C-9),
(C-3), 132.67 (NH–C=O–NH), 132.00 (C-3c), 130.66 (C-
0
24.57 (C-8), 121.70 (C-6 ), 117.97 (C-6), 116.15 (C-10),
7), 124.93 (C-9), 124.63 (C-8), 118.98 (C-4 ), 117.56 (C-6),
0 0 0
0
0
?
15.82 (C-1 ), 115.11 (C-3 ); MS m/z 383.0504 [MH ].
117.46 (C-2), 113.38 (C-1 ), 117.19 (C-9 ), 118.08 (C-11 ),
0 0 0 0
12.60 (C-6 ), 112.34 (C-10 ), 112.21 (C5 ), 111.77 (C-3 ),
1
0
0
?
108.17 (C-8 ), 108.59 (C-12 ); MS m/z 491.0312 [MH ].
N-((2,4-difluorophenyl)carbamoyl)-4-oxo-4H-thieno[3,2-c]chro-
mene-2-carboxamide (8) This compound was synthesized
following the general procedure using 2,4-difluorophenyliso-
Antitumour activity assays
cyanate (0.49 ml) to give 8 (124 mg, 15 %, m.p. =
1
2
59–261 °C). H NMR (DMSO-d ) (d/ppm): 11.72 (1H, s,
Antiproliferative effects of new hydroxamic acid (4 and 5)
and urea (7–11) derivatives containing coumarin[3,2-
c]thiophene moiety were evaluated on human tumour cell
lines including cervical carcinoma (HeLa), colorectal
adenocarcinoma, metastatic (SW620), breast epithelial
adenocarcinoma, metastatic (MCF-7), pancreatic carci-
noma (MiaPaCa-2) and hepatocellular carcinoma (HepG2)
as well as on normal diploid human fibroblasts (BJ). Cells
were cultured as monolayers and maintained in Dulbecco’s
modified Eagle medium (DMEM) supplemented with 10 %
foetal bovine serum (FBS), 2 mM L-glutamine, 100 U/mL
penicillin and 100 lg/mL streptomycin in a humidified
6
NH), 9.75 (1H, s, NH), 8.83 (1H, s, H-3), 8.19–7.03 (7H, m,
13
ar–H); C NMR (DMSO) d/ppm: 166.80 (C=O-2), 160.04
0 0
C-2 ), 158.13 (C-6 ), 156.17 (C-4), 152.61 (C-2c), 150.99 (C-
(
1
1
1
1
1), 142.57 (C-3), 142.00 (C-2), 132.05 (NH–C=O–NH),
31.45 (C-7), 125.35 (C-3c), 125.26 (C-9), 124.16 (C-8),
0
0
23.80 (C-6 ), 121.57 (C-1 ), 117.97 (C-6), 116.60 (C-10),
0
0
?
10.83 (C-5 ), 104.23 (C-3 ); MS m/z 401.0398 [MH ].
N-((2,6-difluorophenyl)carbamoyl)-4-oxo-4H-thieno[3,2-c]chro-
mene-2-carboxamide (9) This compound was synthesized
following the general procedure using 2,6-difluo-
rophenylisocyanate (0.51 ml) to give 9 (117 mg, 14 %,
1
atmosphere with 5 % CO at 37 °C. Cells were plated into
2
m.p. = 263–265 °C). H NMR (DMSO-d ) (d/ppm): 11.68
96-well microtiter plates on day 0 at seeding density of
3000–6000 cells per well depending upon their specific
doubling times. Freshly prepared dilutions of test com-
pounds in culture medium in the concentration range from
6
(
1H, s, NH), 9.89 (1H, s, NH), 8.89 (1H, s, H-3), 8.17–7.20
1
3
(
7H, m, ar–H); C NMR (DMSO) d/ppm: 166.80 (C=O-2),
0
0
1
1
58.78 (C-2 ), 157.13 (C-6 ), 156.08 (C-4), 152.18 (C-2c),
-
8
-4
50.82 (C-11), 141.48 (C-3), 140.90 (C-2), 132.70 (NH–
0
10 to 10 M were added to the wells, and the cells were
grown for further 72 h. The solvent (DMSO) was also
tested for its potential inhibitory activity by adjusting its
C=O–NH), 130.93 (C-7), 128.42 (C-4 ), 125.64 (C-3c),
1
24.67 (C-9), 124.62 (C-8), 117.53 (C-6), 116.21 (C-10),
1
23