898 Devegowda et al.
Asian J. Chem.
m.p.: 154-156 °C. IR (KBr, νmax, cm-1): 3432, 3219, 1595, 1289.
1H NMR (DMSO-d6, 400 MHz) δ: 10.41 (s, 1H, -NH), 9.56
(s, 1H, -NH), 8.49 (s, 1H, Ar-H), 8.22 (s, 1H, Ar-H), 7.95 (s,
1H, Ar-H), 7.80 (m, 1H, Ar-H), 7.61 (m, 2H, Ar-H), 7.52 (d,
J=8.0 Hz, 2H, Ar-H), 7.44 (d, J=8.0 Hz, 2H, Ar-H), 7.18 (d,
1H, Ar-H), 3.97 (s, 3H, -OCH3), 3.95 (s, 3H, -OCH3), 1.32(s,
3.98 (s, 3H, -OCH3), 3.94 (s, 3H, -OCH3). 13C NMR (DMSO-
d6, 100 MHz) δ: 168.4, 158.5, 154.6, 151.3, 146.6, 143.2,
139.7, 137.5, 131.9, 130.3, 129.0, 127.3, 112.8, 109.4, 108.2,
107.8, 102.7, 99.4, 56.2, 56.0. MS (ESI + ion): m/z = 436.7.
Anal. calcd. for C22H20N4O4S: C 60.54, H 4.62, N 12.84. Found:
C 60.59, H 4.66, N 12.89.
13
9H, -(C(CH3)3)). C NMR (DMSO-d6, 100 MHz) δ: 168.4,
N-(3-((6,7-Dimethoxyquinazoline-4yl)amino)phenyl)-
2,3,5-trimethylbenzene sulphonamide (4J): White solid;
m.p.: 166-168 °C. IR (KBr, cm-1): 3466, 3239, 1608, 1277. 1H
NMR (DMSO-d6, 400 MHz) δ: 10.36 (s, 1H, -NH), 9.57 (s,
1H, -NH), 8.46 (s, 1H, Ar-H), 8.27 (s, 1H, Ar-H), 7. 96 (s, 1H,
Ar-H), 7.64 (m, 2H, Ar-H), 7.59 (s, 1H,Ar-H),7.50 (s, 1H, Ar-
H), 7.45 (m, 1H, Ar-H), 7.21 (m, 1H, Ar-H), 2.38-2.42 (s, 9H,
Ar-CH3), 3.96 (s, 3H, -OCH3), 3.93 (s, 3H, -OCH3).13C NMR
(DMSO-d6, 100 MHz) δ: 168.3, 158.4, 154.5, 151.3, 146.6,
143.2, 138.7, 137.5, 137.0, 135.6, 134.0, 132.3, 130.3, 123.6,
112.9, 109.4, 108.2, 107.8, 102.7, 99.4, 56.2, 56.0, 21.6, 19.2,
19.1. MS (ESI + ion): m/z = 479.Anal. calcd. for C25H26N4O4S:
C 62.74, H 5.48, N 11.71. Found: C 62.79, H 5.52, N 11.76.
Experimental procedure for biological assays: Swiss
albino mice were procured from the central animal facility,
Department of studies in Zoology, University of Mysore,
Mysore, India. Ehrlich ascites carcinoma (EAC) cells were
obtained from NCCS (Pune).
158.5, 154.5, 151.3, 146.6, 143.2, 137.5, 136.6, 130.3, 128.1,
125.4, 112.8, 109.3, 107.0, 108.2, 107.8, 102.7, 99.4, 56.2,
56.0, 34.2, 31.3. MS (ESI + ion): m/z = 493.0. Anal. calcd. for
C26H28N4O4S: C 63.40, H 5.73, N 11.37. Found: C 63.45, H
5.78, N 11.42.
N-(3-((6,7-Dimethoxyquinazoline-4-yl)amino)phenyl)-
2-nitrobenzene sulphonamide (4F): White solid; m.p.: 182-
184 °C. IR (KBr, νmax, cm-1): 3441, 3227, 1581, 1283. 1H NMR
(DMSO-d6, 400 MHz) δ: 10.41 (s, 1H, -NH), 9.56 (s, 1H,
-NH), 8.47 (s, 1H, Ar-H), 8.26 (s, 1H, Ar-H), 8.19 (s, 1H, Ar-
H), 7.97 (s, 1H, Ar-H), 7.84 (d, J=8.0 Hz, 2H, Ar-H), 7.76 (s,
1H, Ar-H), 7.62 (m, 1H, Ar-H), 7.46 (m, 2H, Ar-H), 7.39 (m,
1H,Ar-H), 3.99 (s, 3H, -OCH3), 3.96 (s, 3H, -OCH3). 13C NMR
(DMSO-d6, 100 MHz) δ: 168.4, 158.5, 154.6, 151.2, 147.1,
146.5, 143.2, 137.5, 135.1, 134.4, 132.8, 130.3, 128.2, 124.2,
112.9, 109.4, 108.2, 107.8, 102.7, 99.4, 56.2, 56.0. MS (ESI
+ ion): m/z = 481.7. Anal. calcd. for C22H19N5O6S: C 54.88, H
3.98, N 14.55. Found: C 54.93, H 4.04, N 14.61.
Antiproliferative assay: All the newly synthesized
compounds 4(A-J) were preliminarily evaluated for their anti-
proliferative activity on EAC cell lines using MTT assay [25].
Ehrlich ascites carcinoma cell lines (Mouse mammary carcinoma)
were cultured in modified eagle’s medium (MEM), supple-
mented with 10 % FBS at 37 °C and humidified atmosphere
containing 5 % CO2. Cells were seeded at a density of 1 × 104
cells/well in a 96 well plate and allowed to attach for 24 h.
The media was removed and treated with different concen-
trations (1-100 µg) of compound for 24 h. Then, 10 µL of
MTT solution (5 mg/mL) was added to each well and incubated
at 37 °C for 4 h. Then, medium was removed and 200 µL of
dimethyl sulfoxide was added to each well in order to solubilize
formazan crystals. The absorbance was measured at 570 nm by
micro plate reader. The IC50 value was defined as the concen-
tration that caused 50 % inhibition of cell proliferation.
in vivoTreatment of compound 4G: Ehrlich ascites carci-
noma (5 × 106 cells/mouse) cells were injected intraperitonially
into six to eight weeks old Swiss albino mice and the animals
showed a significant increase in the body weight over the
growth period. To determine, whether the compound inhibit
tumor growth, compound 4G (100 mg/kg) was administered
into the EAC bearing mice on every alternate day starting from
6th day of inoculation. The growth of tumor was monitored by
taking the body weight every day. The animals were sacrificed
on 11th day. The volume of ascites from both treated and control
were noted. The other group animals were used to study the
survivability after treatment until their death [26].
N-(3-((6,7-Dimethoxyquinazoline-4-yl)amino)phenyl)-
4-nitrobenzene sulphonamide (4G): White solid; m.p.: 196-
198 °C. IR (KBr, νmax, cm-1): 3449, 3235, 1585, 1288. 1H NMR
(DMSO-d6, 400 MHz) δ: 10.43 (s, 1H, -NH), 9.60 (s, 1H,
-NH), 8.53 (s, 1H, Ar-H), 8.22 (s, 1H, Ar-H), 7.94 (s, 1H, Ar-
H), 7.82 (m, 1H, Ar-H), 7.63 (d, J=8.0 Hz, 2H, Ar-H), 7.54
(m, 2H, Ar-H), 7.45 (d, J=8.0 Hz, 2H, Ar-H), 7.23 (m, 1H, Ar-
H), 3.96 (s, 3H, -OCH3), 3.94 (s, 3H, -OCH3). 13C NMR
(DMSO-d6, 100 MHz) δ: 168.4, 158.5, 154.6, 151.3, 151.1,
146.6, 145.8, 143.2, 137.5, 130.3, 128.2, 124.2, 112.9, 109.4,
108.2, 107.8, 102.7, 99.4, 56.2, 56.0. MS (ESI + ion): m/z =
481.7.Anal. calcd. for C22H19N5O6S: C 54.88, H 3.98, N 14.55.
Found: C 54.94, H 4.06, N 14.60.
N-(3-((6,7-Dimethoxyquinazoline-4-yl)amino)phenyl)-
3-nitrobenzene sulfoamide (4H): White solid; m.p.: 190-192
°C. IR (KBr, cm-1): 3452, 3231, 1596, 1293. 1H NMR (DMSO-
d6, 400 MHz) δ: 10.39 (s, 1H, -NH), 9.57 (s, 1H, -NH), 8.46
(s, 1H, Ar-H), 8.23 (s, 1H, Ar-H), 8.11 (s, 1H, Ar-H), 7.93 (s,
1H, Ar-H), 7.81 (m, 2H, Ar-H), 7.77 (s, 1H, Ar-H), 7.56 (m,
1H, Ar-H), 7.43 (m, 2H, Ar-H), 7.49 (m, 1H, Ar-H), 3.97 (s,
3H, -OCH3), 3.94 (s, 3H, -OCH3). 13C NMR (DMSO-d6, 100
MHz) δ: 168.4, 158.5, 154.6, 151.3, 148.3, 146.6, 143.2, 140.6,
137.5, 133.4, 130.3, 129.9, 127.1, 123.1, 112.9, 109.4, 108.2,
107.8, 102.7, 99.4, 56.2, 56.0. MS (ESI + ion): m/z = 481.7.
Anal. calcd. for C22H19N5O6S: C 54.88, H 3.98, N 14.55. Found:
C 54.92, H 4.02, N 14.58.
N-(3-((6,7-Dimethoxyquinazoline-4-yl)amino)phenyl)-
benzenesulfonamide (4I): White solid; m.p.: 148-150 °C. IR
(KBr, νmax, cm-1): 3456, 3241, 1602, 1282. 1H NMR (DMSO-
d6, 400 MHz) δ: 10.33 (s, 1H, -NH), 9.53 (s, 1H, -NH), 8.44
(s, 1H, Ar-H), 8.21 (s, 1H, Ar-H), 8.12 (s, 1H, Ar-H), 7.93 (s,
1H, Ar-H), 7.82 (d, J=8.0 Hz, 2H, Ar-H), 7.74 (s, 1H, Ar-H),
7.60 (m, 2H, Ar-H), 7.40 (m, 2H, Ar-H), 7.33 (m, 1H, Ar-H),
Studies on cell morphology: Giemsa and nuclear staining
were performed and visualized using light and fluorescent
(Leitz-DIAPLAN) microscope. Both compound 4G treated
and untreated cells were harvested from mice, fixed in methanol:
acetic acid (3:1) and were smeared on glass slide and air-dried
in humidified chamber. The cells were hydrated with PBS and