282
S. Sestito et al. / European Journal of Medicinal Chemistry 105 (2015) 274e288
Fig. 4. ANGM-CSS-derived GSCs were incubated in Neural Stem Cells (NSC) medium with the indicated concentrations of 3 for three (A) or seven days (A, B). At the end of the
treatment periods, cell proliferation was measured using the MTS assay. The data are expressed as a percentage with respect to that of untreated cells (control), and are the mean
values ± SEM of three independent experiments, each performed in duplicate. IC50 values after 7 days of treatment were calculated from sigmoid doseeresponse curve (B). The
significance of the differences was determined with a one-way ANOVA with Bonferroni post-test: ***p < 0.001 vs. control.
4.2.1. (3Z)-2-((3,4-dimethoxybenzyl)amino)-N-(2-oxo-3-
(C21
H
19
N
5
O
2
S) Calc%: C 62.21, H 4.72, N 17.27, S 7.91. Found%: C
(
thiophen-2-ylmethylene) indolin-5-yl)acetamide (4)
62.35, H 4.89, N 17.03, S 8.03.
The residual material was purified by crystallization from
AcOEt/n-hexane, affording 4 as the Z-isomer (36 mg, 0.08 mmol,
0
4.2.4. (3Z)-N-(4-(methylthio)phenyl)-N -(2-oxo-(3-(1H-imidazol-
5-yl)methylene)indolin-5-yl)urea (7)
ꢃ
1
2
1
CH
CH
0% yield): mp 167e169 C. H NMR (CDCl3):
d
3.37 (dd, 1H, J ¼ 1.8;
5.1 Hz, CH
2
); 3.55 (d, 1H, J ¼ 12.9 Hz, CH
2
); 3.75 (d, 1H, J ¼ 15.1 Hz,
The residual material was purified by crystallization from EtOH,
affording 7 as the Z-isomer (98 mg, 0.25 mmol, 63% yield): mp
2
); 3.86 (s, 3H, OMe); 3.89 (s, 3H, OMe); 4.00 (d, 1H, J ¼ 12.9 Hz,
ꢃ
1
2
); 6.66 (d, 1H, J ¼ 8.2 Hz, Ar); 6.79e6.96 (m, 3H, Ar); 7.11 (dd,1H,
215e217 C. H NMR:
d
2.44 (s, 3H, SMe); 6.82 (d, 1H, J ¼ 8.3 Hz, Ar);
J ¼ 3.6, 5.1 Hz, Ar); 7.32e7.35 (m, 1H, Ar); 7.58 (s, 1H, H-vinyl); 7.62
7.12 (dd, 1H, J ¼ 1.3, 8.3 Hz, Ar); 7.22 (d, 2H, J ¼ 8.6 Hz, Ar); 7.44 (d,
2H, J ¼ 8.6 Hz, Ar); 7.71 (s, 1H, H-vinyl); 7.79 (s, 1H, Ar); 7.86 (d, 1H,
J ¼ 1.3 Hz, Ar); 8.02 (s, 1H, Ar); 8.54 (br s, 1H); 8.70 (br s, 1H); 10.90
(
d, 1H, J ¼ 5.1 Hz); 7.76 (d, 1H, J ¼ 3.6 Hz, Ar); 8.00 (s, 1H, Ar) ppm.
1
3
C NMR (CDCl3):
37.03, 133.48, 128.95, 128.94, 128.93, 128.16, 126.94, 126.09, 124.97,
20.22, 116.49, 111.23, 110.63, 109.22, 78.63, 55.49, 54.44 ppm. Anal.
S) Calc%: C 64.13, H 5.16, N 9.35, S 7.13. Found%: C
4.25, H 5.27, N 9.58, S 7.25.
d 169.84, 167.85, 148.76, 148.14, 141.84, 137.67,
13
1
1
(br s, 1H, NH) ppm. C NMR: d 167.58,153.00,138.60,137.01,135.82,
134.00, 133.27, 128.54, 127.41, 127.03 124.93, 122.07, 120.49, 120.01,
118.58, 111.35, 109.15; 17.18 ppm. Anal. (C20H17N O S) Calc%: C
24 23 3 4
(C H N O
5
2
6
61.37, H 4.38, N 17.89, S 8.19. Found%: C 61.45, H 4.30, N 18.01, S 8.23.
4.3. General procedure for the synthesis of compounds 8-10
A solution of oxone (215 mg, 0.35 mmol) in water was added to a
0
.2.2. (Z)-N-(4-(methylthio)phenyl)-N -(2-oxo-3-(thiophen-2-
ylmethylene)indolin-5-yl)urea (5)
The residual material was purified by crystallization from EtOH,
affording 5 as the Z-isomer (94 mg, 0.23 mmol, 60% yield): mp
4
ꢃ
cooled (0 C) solution of the appropriate derivative 8e10
ꢃ
1
2
46e248 C. H NMR:
Ar); 7.11 (d, 1H, J ¼ 8.2 Hz, Ar); 7.22 (d, 2H, J ¼ 8.4 Hz, Ar); 7.44 (d,
H, J ¼ 8.4 Hz, Ar); 7.84e7.92 (m, 3H, Ar, H-vinyl); 7.96e8.01 (m, 1H,
Ar); 8.03 (s, 1H, Ar); 8.48 (br s, 1H); 8.68 (br s, 1H); 10.49 (br s, 1H,
NH) ppm. 13C NMR:
167.16, 152.65, 137.53, 137.11, 135.69, 133.93,
33.16, 129.54, 127.81, 127.16, 124.93, 121.87, 120.71, 119.91, 118.78,
11.17, 110.27, 109.27, 16.10 ppm. Anal. (C21 ) Calc%: C
1.89, H 4.20, N 10.31, S 15.74. Found%: C 61.74, H 4.27, N 10.02, S
d
2.44 (s, 3H, SMe); 6.78 (d, 1H, J ¼ 8.2 Hz,
(0.29 mmol) in 1:1 MeOH-THF (2 mL). The resulting mixture was
stirred at room temperature for 12 h. Then the solid was filtered off,
and the solution was evaporated. The residual material was diluted
with MeOH and the solid product was separated out from the
2
d
mixture. It was filtered, washed with Et
2
O, and air dried.
1
1
6
1
0
H
17
N
3
O
2
S
2
4.3.1. (3Z)-N-(4-(methylsulfonyl)phenyl)-N -(2-oxo-3-(thiophen-2-
ylmethylene)indolin-5-yl)urea (8)
ꢃ
1
5.55.
(76 mg, 0.17 mmol, 60% yield): mp 213e215 C. H NMR: d 3.16
(
s, 3H, SMe); 6.79 (d, 1H, J ¼ 8.3 Hz, Ar); 7.14 (d, 1H, J ¼ 8.3 Hz, Ar);
0
4
.2.3. (3E)-N-(4-(methylthio)phenyl)-N -(2-oxo-(3-(1-methyl-1H-
7.23 (t, 1H, J ¼ 4.3 Hz, Ar); 7.60e7.92 (m, 6H, Ar); 7.98e8.10 (m, 2H,
imidazol-2-yl)methylene)-indolin-5-yl)urea (6)
The residual material was purified by crystallization from EtOH,
affording 6 as the E-isomer (101 mg, 0.25 mmol, 64% yield): mp
Ar, H-vinyl); 8.75 (br s,1H); 9.29 (br s,1H); 10.52 (br s,1H, NH) ppm.
1
3
C NMR:
129.54, 127.75, 127.14, 125.03, 121.87, 120.98, 118.94, 118.75, 111.17,
110.27, 109.27; 52.70 ppm. Anal. (C21 ) Calc%: C 57.39, H
d 168.40, 152.56, 148.00, 137.71, 136.09, 134.12, 133.18,
ꢃ
1
2
35e237 C. H NMR:
d
2.43 (s, 3H, SMe); 3.90 (s, 3H, NMe); 6.78 (d,
17 3 4 2
H N O S
1
H, J ¼ 8.3 Hz, Ar); 7.21 (d, 2H, J ¼ 8.8 Hz, Ar); 7.36 (s, 2H, Ar); 7.42
3.90, N 9.56, S 14.59. Found%: C 57.54, H 4.01, N 10.10, S 14.78.
(
8
1
d, 2H, J ¼ 8.8 Hz, Ar); 7.53 (s, 1H, H-vinyl); 7.65 (dd, 1H, J ¼ 2.0,
.3 Hz, Ar); 8.49 (br s,1H); 8.64 (br s,1H); 9.12 (d,1H, J ¼ 2.0 Hz, Ar);
0
4.3.2. (3E)-N-(4-(methylsulfonyl)phenyl)-N -(2-oxo-3-((1-methyl-
1
3
0.47 (br s, 1H, NH) ppm. C NMR:
37.00, 138.86, 133.66, 130.76, 129.42, 128.27, 127.52, 123.70, 122.84,
21.55, 119.78, 112.57, 108.97; 34.25, 14.20 ppm. Anal.
d
168.27, 157.45, 140.26, 139.14,
1H-imidazol-2-yl)methylene)indolin-5-yl)urea (9)
ꢃ
1
1
1
(79 mg, 0.18 mmol, 62% yield): mp 229e231 C. H NMR:
d 3.15 (s,
3H, SMe); 3.90 (s, 3H, NMe); 6.81 (d, 1H, J ¼ 8.2 Hz, Ar); 7.37 (s, 2H,