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Organic & Biomolecular Chemistry
Page 7 of 8
DOI: 10.1039/C6OB02181F
Journal Name
ARTICLE
Subsequently, the cells were treated with 5 mg / mL MTT (40
Compound
2, 4-((E)-4-(9H-carbazol-9-yl)styryl)-N-(pyridin-3-
µL/well) and incubated for an additional 4 h (37 ℃, 5% CO2). ylmethylene) aniline, yellow powder, 0.46 g, yield: 74 %,
°
Then, DMEM was removed, the formazan crystals were melting point: 186.1-187.2 C. FT- IR (ATR, cm−1): 1625.21 (w),
dissolved in DMSO (150 µL/well), and the absorbance at 570 1595.16 (w), 1514.53 (m), 1478.98 (m), 1451.01 (s), 1335.15
nm was recorded. The cell viability (%) was calculated (m), 1316.23 (m), 1231.16 (m), 1183.27 (m), 1119.07 (m),
1
1016.23 (m), 991.81 (m), 833.42 (m), 747.91 (s), 722.00 (s). H
according to the following Equation:
Cell Viability% = OD570 (sample) / OD570 (control) × 100
NMR (400 MHz, DMSO-d6, δ): 9.09 (1 H, s), 8.81 (1 H, s), 8.73 -
Where OD570 (sample) represents the optical density of the 8.72 (1 H, d, J = 4.0), 8.36 - 8.34 (1 H, d, J = 8.0), 8.28 - 8.26 (2 H,
wells treated with various concentration of the compounds d, J = 8.0), 7.93 - 7.91 (2 H, d, J = 8.0), 7.77 - 7.75 (2 H, d, J =
and OD570 (control) represents that of the wells treated with 8.0), 7.65 - 7.63 (2 H, d, J = 8.0), 7.56 - 7.53 (1 H, m), 7.45 - 7.39
DMEM + 10% fetal calf serum (FCS). Three independent trials (8 H, m), 7.31 -7.29 (2 H, m). 13C NMR (100 MHz, DMSO-d6, δ):
were conducted, and the averages and standard deviations are 158.13, 154.57, 151.94, 150.45, 139.98, 136.37, 135.91, 135.41,
reported. The reported percent cell survival values are relative 134.96, 131.58, 128.01, 127.36, 126.27, 124.08, 122.74, 121.74,
to untreated control cells.
120.53, 120.10, 113.93, 109.70. MS (ESI) m/z: found, [M+H]+,
450.1987; molecular formula C32H23N3 requires [M+H]+,
450.1971.
4 Cell Image
HepG2 cells were seeded and grown in 6 well plates at a
density of 2×105 cells per well for 96 h. For live cell imaging,
cell cultures were incubated with the complexes (PBS : cell
media = 1 : 9) at concentrations 20 μM and maintained at
37 °C in an atmosphere of 5% CO2 and 95% air for incubation
times ranging for 2 h. Then the cells were washed with PBS (3
× 3 mL per well) and 3 mL of PBS was added to each cell. The
cells were imaged using confocal laser scanning microscopy
and water immersion lenses, then the fluorescence emission
measured at 520-560 nm with optimal excitation energy.
5. Synthesis and characterization
Compound 3, 4-((E)-4-(9H-carbazol-9-yl)styryl)-N-benzylide-
-neaniline, deep yellow powder,0.40 g, yield: 64 %, melting
°
point: 233.3-234.6 C. FT- IR (ATR, cm−1): 1594.96 (m), 1512.72
(m), 1478.12 (w), 1449.34 (s), 1408.15 (w), 1334.77 (w),
1315.75 (m), 1227.11 (s), 1102.84 (w), 961.86 (m), 839.33 (s),
1
748.65 (s), 723.67 (s). H NMR (400 MHz, DMSO-d6, δ): 8.71 (1
H, s), 8.27-8.25 (2 H, d, J = 8.0), 8.03 - 7.87 (4 H, m), 7.81 - 7.21
(17 H, m). 13C NMR (100 MHz, DMSO-d6, δ): 160.38, 150.76,
140.05, 128.91, 128.68, 128.69, 128.05, 127.69, 127.17, 126.83,
126.27, 122.91, 121.67, 120.57, 120.06, 109.71. MS (APCI) m/z:
found, [M+H]-, 449.2031; molecular formula C32H23N3 requires
[M+H]-, 449.2018.
5.1. Synthesis of the intermediate 1-4
Intermediate
intermediate , were synthesized according to the methods
reported in the literature 12 with some modifications.
.2. Synthesis of Compounds 1 - 6
Intermediate (0.5 g, 1.4 mmol) were dissolved in 60 mL of
1, intermediate 2, intermediate 3 and
Compound 4, 2-(((4-((E)-4-(9H-carbazol-9-yl)styryl)phenyl)i-
4
-mino)methyl) phenol, yellow powder, 0.43 g, yield: 67 %,
°
melting point: 251.2-252.2 C. FT- IR (ATR, cm−1): 3023.25 (w),
5
1594.96 (m), 1512.72 (m), 1478.12 (m), 1449.34 (s), 1334.77
(m), 1227.11 (m), 1183.33 (w), 961.86 (m), 839.33 (s), 748.65
(s), 723.67 (s). (0.43 g, yield: 66.7 %,): 1H NMR (400 MHz,
DMSO-d6, δ): 13.13 (1H, s), 9.04 (1H, s), 8.27-8.25 (2H, d, J =
8.0), 7.93-7.91 (2H, d, J = 8.0), 7.79-7.77 (2H, d, J = 8.0), 7.69-
7.66 (3H, dd, J = 12.0), 7.55 - 7.37 (9H, m), 7.41 -7.30 (2H, m),
7.00 - 6.98 (2H, m). 13C NMR (100 MHz, DMSO-d6, δ): 163.45,
160.82, 156.94, 147.76, 140.52, 132.97, 130.54, 129.10, 128.61,
128.21, 127.25, 126.69, 123.23, 122.40, 121.06, 120.59, 119.95,
119.61, 117.09, 110.11. MS (ESI) m/z: found, [M+H]+, 465.1980;
molecular formula C33H24N2O requires [M+H]+, 465.1968.
4
methanol and stirred for 10 min. Then glacial acetic acid was
added in catalytic amount (3 drops) and corresponding
aldehyde (1.8 mmol) was added dropwise to the reaction
°
system, the reaction mixture was refluxed at 65 C for 2 h.
After the reaction finished, the mixture was immediately
filtered under reduced pressure and recrystallization used
methanol to obtain pure product.
Compound 1, 4-((E)-4-(9H-carbazol-9-yl)styryl)-N-(pyridin-2-
ylmethylene) aniline, yellow powder, 0.41 g, yield: 59 %,
°
melting point: 206.5-207.4 C. FT- IR (ATR, cm−1): 1595.48 (w),
Compound 5, 4-(((4-((E)-4-(9H-carbazol-9-yl)styryl)phenyl)i-
1515.26 (m), 1479.07 (w), 1451.52 (s), 1435.11 (m), 1365.23
(m), 1335.10 (m), 1316.28 (m), 1230.84 (s), 1184.19 (w),
-mino)methyl)-N,N-diethylaniline, yellow powder, 0.50 g, yield:
69 %, melting point: 209.1-211.4 °C. FT- IR (ATR, cm−1): 3023.25
(w), 1616.36 (m), 1596.10 (m), 1514.46 (m), 1450.03 (s),
1359.65 (m), 1334.89 (m), 1279.58 (m), 1226.06 (m), 1170.21
(m), 1152.14 (m), 964.96 (m), 835.66 (m), 746.93 (s), 721.95 (s).
1H NMR (400 MHz, DMSO-d6, δ): 8.45 (1 H, s), 8.27 - 8.25 (2 H,
d, J = 8.0), 7.91-7.89 (2 H, d, J = 8.0), 7.75-7.73 (2 H, d, J = 8.0),
7.69-7.63 (4 H, dd, J = 8.0, 8.0), 7.74 - 7.24 (10 H, m), 6.77-6.75
(2 H, d, J = 8.0), 3.46-3.41 (4 H, m), 1.16-1.13 (6 H, t, J = 8.0).
13C NMR (100 MHz, CD3COCD3, δ): 159.31, 131.33, 130.21,
129.65, 128.24, 127.87, 127.23, 126.83, 126.23, 121.62, 114.58,
110.81, 110.11, 44.19, 11.65. MS (ESI) m/z: found, [M+H]+,
520.2767; molecular formula C37H33N3 requires [M+H]+,
520.2753.
1
950.82 (w), 832.65 (m), 746.57 (s), 723.20 (s), 623.81 (m). H
NMR (400 MHz, DMSO-d6, δ): 8.75 - 8.74 (1 H, d, J = 4.0), 8.68
(1 H, s), 8.27 - 8.26 (2 H, d, J = 4.0), 8.20 -8.18 (1 H, d, J = 8.0),
8.00 - 7.96 (1 H, t, J = 8.0, 8.0), 7.94 - 7.91 (2 H, d, J = 12.0),
7.77 - 7.75 (2 H, d, J = 8.0), 7.67 - 7.65 (2 H, d, J = 8.0), 7.56 -
7.54 (1 H, m), 7.45 - 7.43 (8 H, m), 7.32 - 7.29 (2 H, m). 13C
NMR (100 MHz, DMSO-d6, δ): 160.33, 154.06, 149.72, 139.98,
137.07, 135.94, 135.76, 127.62, 127.52, 126.81, 126.27, 125.65,
121.84, 121.23, 120.53, 120.10, 113.83, 109.70. MS (ESI) m/z:
found, [M+H]+, 450.1981; molecular formula C32H23N3 requires
[M+H]+, 450.1971.
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