4
RAHMANNEJADI ET AL.
2
1
1
1
7
8.1, 28.3, 30.5, 34.4, 45.4, 48.2, 69.3, 106.3, 114.3, 115.9,
17.9, 121.7, 127.2, 128.5, 128.6, 135.1, 136.6, 145.2,
46.7, 156.0, 160.0, 160.9 ppm; IR (KBr): = 3232 (N–H),
1.70–1.72 (m, 2H, CH ), 1.78–1.81 (m, 2H, CH ), 2.57–
2
2
2.61 (m, 2H, CH ), 6.60 (d, J = 8.5 Hz, 1H, H–Ar), 6.91
2
(s, 1H, NH), 7.31 (d of d, J = 2.0, 9.0 Hz, 1H, H–Ar),
7.54 (d, J = 8.5 Hz, 1H, H–Ar), 7.59 (d, J = 2.0 Hz, 1H,
H–Ar), 7.93 (d of d, J = 2.0, 9.0 Hz, 1H, H–Ar), 8.12 (s,
1H, NH), 8.15 (d, J = 2.0 Hz, 1H. H‐Ar), 12.3 (br s, 1H,
−
1
676 (C=O), 1610 (C=N), 1249 (C–N) cm ; MS (EI,
+
0 eV): m/z (%) = 534 (M , 4), 279 (27), 239 (52), 198
(38), 161 (29), 77 (100).
13
NH) ppm; C NMR (125 MHz, DMSO‐d ): δ = 20.9,
6
2
1
1
1
7.3, 33.3, 39.6, 68.5, 106.4, 114.6, 115.9, 118.2, 121.7,
24.7, 127.4, 128.3, 128.6, 135.1, 136.8, 145.5, 158.3,
59.8, 161.2 ppm; IR (KBr): = 3317 (N–H), 1672 (C=O),
3
.3 | 6,8‐Dibromo‐2‐[3‐(6,8‐dibromo‐2‐
methyl‐4‐oxo‐1,2,3,4‐tetrahydro‐quinazolin‐
2
one (3c)
‐yl)‐2,2‐dimethyl propyl]‐3H‐quinazolin‐4‐
−1
607 (C=N), 1254 (C–N) cm .
Pale yellow solid; Yield 0.387 g (56%); M.p.: 300°C
3.6 | 6,8‐Dibromo‐2‐[3‐(6,8‐dibromo‐2‐
methyl‐4‐oxo‐1,2,3,4‐tetrahydro‐quinazolin‐
2‐yl)‐propyl]‐3H‐quinazolin‐4‐one (3f)
1
(
(
decomposed); H NMR (500 MHz, DMSO‐d ): δ = 1.03
6
s, 3H, Me), 1.12 (s, 3H, Me), 1.53 (s, 3H, Me), 1.77 (AB‐
2
quartet, ΔvAB = 55 Hz, J = 15.0 Hz, 2H, CH ), 2.64
AB
2
2
Pale yellow solid; Yield 0.526 g (79%); M.p.: 300°C
(AB‐quartet, ΔvAB = 34 Hz, J = 13.0 Hz, 2H, CH2),
AB
1
(
(
decomposed); H NMR (500 MHz, DMSO‐d ): δ = 1.45
6
8
1
.66 (s, 1H, N–H), 7.63 (s, 1H, H–Ar), 7.69 (s, 1H, H–Ar),
6
s, 3H, Me), 1.75–1.77 (m, 2H, CH ), 1.89–1.94 (m, 2H,
.17 (s, 1H, H–Ar), 8.32 (s, 1H, H–N), 8.44 (s, 1H, H–Ar),
2
13
CH ), 2.58–2.60 (m, 2H, CH ), 6.23 (s, 1H, NH), 7.65 (d
2.63 (s, 1H, H–N) ppm; C NMR (125 MHz, DMSO‐d6):
2
2
of d, J = 2.0, 8.0 Hz, 2H, H–Ar), 8.12 (d, J = 2.5 Hz, 1H,
H–Ar), 8.26 (d, J = 2.5 Hz,1H, H–Ar), 8.37 (br s, 1H,
δ = 29.7, 30.0, 32.0, 35,0, 44.7, 46.6, 71,0, 107.2, 108.6,
1
1
1
16.4, 118.8, 123.1, 123.9, 128.2, 129.5, 138.2, 140.1, 143.3,
13
NH), 12.57 (br s, 1H, NH) ppm; C NMR (125 MHz,
45.5, 158.3, 160.6, 160.9 ppm; IR (KBr): = 3337 (N–H),
−
1
DMSO‐d ): δ = 20.7, 27.2, 33.8, 39.5, 69.2, 106.5, 107.6,
646 (C=O), 1605 (C=N), 1241 (C–N) cm .
6
1
1
16.2, 117.4, 122.4, 122.9, 127.1, 128.5, 137.1, 138.8,
42.8, 144.9, 158.3, 159.8, 160.2 ppm; IR (KBr): = 3397
−
1
3
.4 | 2‐[3‐(2‐Methyl‐4‐oxo‐1,2,3,4‐
(N–H), 1678 (C=O), 1614 (C=N), 1261 (C–N) cm ; MS
(EI, 70 eV): m/z (%) = 665 [(M + 1) , 4], 664 (M , 1),
+
+
tetrahydro‐quinazolin‐2‐yl)‐propyl]‐3H‐
quinazolin‐4‐one (3d)
331 (25.6), 319 (100), 277 (26), 238 (16).
1
Pale yellow solid; Yield 0.245 g (70%); M.p.: 210–212°C; H
3.7 | MTT assay
NMR (500 MHz, DMSO‐d ): δ = 1.36 (s, 3H, Me), 1.71–1.74
6
(
m, 2H, CH ), 1.84–1.90 (m, 2H, CH ), 2.59–2.62 (m, 2H,
2
2
The cytotoxic effects of bis‐quinazoline compounds were
CH ), 6.58–6.61 (t, J = 7.5 Hz, 1H, H–Ar), 6.64 (s, 1H,
NH), 6.66 (s, 1H. NH), 7.11–7.14 (m, 1H, H–Ar), 7.48 (t,
J = 7.5 Hz, 1H, H–Ar), 7.57 (d, J = 8.0 Hz, 1H, H–Ar),
2
investigated using a standard MTT assay. Briefly the cells
3
were plated at the 8 × 10 cells concentration for A549
3
and 10 × 10 cells concentration for SKOV3 and MCF‐7
7
.61 (d, J = 8.0 Hz, 1H, H–Ar), 7.79 (t, J = 7.5 Hz, 1H, H–
3
in 100 mm complete culture media per well. Then, the
Ar), 7.96 (s, 1H, NH), 8.10 (d, J = 7.5 Hz, 1H, H–Ar) ppm;
1
3
cells were incubated for 24 h to reattach. After attachment
the cells were treated with different concentrations of each
compound (1–500 μM). All compound were prepared in
DMSO (Sigma, Germany) as 400 μM as stock solution.
The desired concentrations were then obtained by serial
dilution of stocks in culture media. The final concentration
of DMSO was kept to less than 10% to avoid solvent cyto-
toxic effect. Three wells were left without treatment as
cell‐based negative controls, and three wells containing
cell culture medium alone were considered as blanks. After
72 hours incubation, the culture media were completely
removed and 100 μL of MTT solution with 0.5 mg/mL con-
centration were added to the wells including controls. The
plate was incubated for 3–4 h at 37°C and checked period-
ically for the appearance of purple precipitate. Then, after
C NMR (125 MHz, DMSO‐d ): δ = 21.2, 27.4, 33.3, 39.7,
6
6
1
=
8.5, 113.0, 113.5, 115.7, 120.0, 125.4, 125.9, 126.6, 127.6,
28.3, 132.7, 134.1, 146.6, 157.9, 160.8, 162.6 ppm; IR (KBr):
−
1
3242 (N–H), 1680 (C=O), 1612 (C=N), 1277 (C–N) cm ;
+
+
MS (EI, 70 eV): m/z (%) = 349 [(M + 1) , 3], 348 (M , 1),
3
6
31 (33), 247 (5), 173.1 (35), 161 (100), 128 (30),77 (19),
9 (58).
3
.5 | 6‐Bromo‐2‐[3‐(6‐bromo‐2‐methyl‐4‐
oxo‐1,2,3,4‐tetrahydro‐quinazolin‐2‐yl)‐
propyl]‐3H‐quinazolin‐4‐one (3e)
Pale yellow solid; Yield 0.313 g (62%); M.p.: 230–232°C;
1
H NMR (500 MHz, DMSO‐d ): δ = 1.35 (s, 3H, Me),
6