L.B. Salum et al. / European Journal of Medicinal Chemistry 63 (2013) 501e510
503
NMR spectra revealed that structures were geometrically pure and
configured E (JH
¼ w16 Hz), except 3a, which is a mixture of E
2.1.2.5. 3q e (2E)-1-(30,40,50-trimethoxyphenyl)-3-(3-trifluoromethyl-
4-chlorophenyl)-2-propen-1-one. Light yellow solid, m.p. 146e
a
ꢁH
b
(JH
¼ 16 Hz) 2:1 Z (JH
¼ 12 Hz) isomers.
147 ꢂC; 1H NMR (CDCl3)
d 3.95 (s, 3H, p-OCH3), 3.95 (s, 6H, m-
a
ꢁHb
a
ꢁHb
OCH3), 7.26 (2, 2H, H20, H60), 7.32 (dd, 1H, J ¼ 8.0/1.0 Hz, H5), 7.38 (d,
2.1.2.1. 2c e (2E)-1-(30,40,50-trimethoxyphenyl)-3-(2-naphthyl)-2-
propen-1-one. Light yellow solid, m.p. 131e132 ꢂC; 1H NMR
1H, J ¼ 16.0 Hz, H ), 7.48 (s, 1H, H2), 7.68 (d, 1H, J ¼ 8.0 Hz, H6), 8.08
a
(d, 1H, J ¼ 16.0 Hz, H
OCH3), 106.49 (C20, C60), 125.32 (C
(C3), 130.38 (C6), 132.13 (C4), 133.17 (C10), 136.19 (CF3), 136.70 (C1),
b
). 13C NMR (CDCl3)
), 127.80 (C2), 128.78 (C5), 130.34
d 56.61 (m-OCH3), 61.16 (p-
(CDCl3)
7.54 (m, 2H, H3, H6), 7.60 (d, 1H, J ¼ 15.6 Hz, H
J ¼ 8.0 Hz, H7), 7.87 (m, 2H, H5, H8), 7.89 (m, 1H, H4), 7.99 (d, 1H,
J ¼ 15.6 Hz, H 56.66 (m-OCH3),
), 8.05 (s, 1H, H1). 13C NMR (CDCl3)
61.28 (p-OCH3), 106.29 (C20, C60), 122.04 (C3), 123.86 (C
), 127.07
(C1), 127.68 (C6), 128.07 (C7), 128.90 (C5), 128.98 (C8), 130.99 (C4),
d
3.96 (s, 6H, OCH3), 3.98 (s, 3H, OCH3), 7.32 (s, 2H, H20, H60),
a
a
), 7.83 (d, 1H,
139.53 (Cb /
), 141.59 (C40), 153.41 (C30, C50), 189.40 (C]O). IR nmax
b
d
cmꢁ1 1658 (C]O), 1574 (C]C), 1230, 1001 (CeO), 1161 (CeF), 1125
(CeCl), 3080, 2938, 2838, 1598, 1505, 1451, 1413, 1343, 1314, 1104,
816, 648 (Ar) (KBr). Anal. Calcd for C19H16ClF3O4: C 56.94, H 4.02.
Found: C 57.20, H 4.16. Yield: 90%.
a
132.59 (C10), 133.59 (C2),133.84 (C4a), 134.61 (C8a), 145.14 (C40, C
b),
153.40 (C30, C50),189.47 (C]O). IR nmax/cmꢁ11656 (C]O),1582 (C]
C), 1230 (CeO), 2834, 1502, 1458, 1411, 996, 813 (Ar) (KBr). Anal.
Calcd for C22H20O4: C 75.84, H 5.79. Found: C 75.53, H 5.81.
Yield ¼ 97%.
2.1.2.6. 3s e (2E)-1-(30,40,50-trimethoxyphenyl)-3-(3,5-
dichlorophenyl)-2-propen-1-one. Cream solid, m.p. 136e138 ꢂC; 1H
NMR (CDCl3)
d 3.95 (s, 3H, OCH3), 3.96 (s, 6H, OCH3), 7.27 (2, 2H,
H20, H60), 7.40 (m, 1H, H4), 7.46 (d, 1H, J ¼ 16.0 Hz, H
a
), 7.50e7.51 (s,
56.74
), 127.29
(C4), 132.38 (C10, C2, C6), 133.16 (C3, C5), 134.10 (C1), 141.79 (C
b),
2.1.2.2. 2d e (2E)-1-(10,30-benzodioxol-5-yl)-3-(2,3,4-
trimethoxyphenyl)-2-propen-1-one. Light yellow solid, m.p. 100e
2H, H2, H6), 7.66 (d, 1H, J ¼ 16.0 Hz, H
b d
). 13C NMR (CDCl3)
(m-OCH3), 61.74 (p-OCH3), 106.44 (C20, C60), 123.85 (C
a
101 ꢂC; 1H NMR (CDCl3)
d
3.89 (s, 3H, OCH3), 3.90 (s, 3H, OCH3),
3.94 (s, 3H, OCH3), 6.05 (s, 2H, eOCH2Oe), 6.71 (d, 1H, J ¼ 8.0 Hz,
143.16 (C40), 153.47 (C30, C50), 188.66 (C]O). IR nmax/cmꢁ1 1664 (C]
O), 1584 (C]C), 1230, 1003 (CeO), 1132 (CeCl), 3069, 2945, 2831,
1610, 1564, 1508, 1467, 1418, 1340, 1164, 973, 838, 803, 660 (Ar)
(KBr). Anal. Calcd for C18H16Cl2O4: C 58.87, H 4.39. Found: C 58.61, H
5.00. Yield: 92%.
H5), 6.88 (d, 1H, J ¼ 8.0 Hz, H50), 7.36 (d, 1H, J ¼ 8.0 Hz, H6), 7.51 (d,
1H, J ¼ 16.0 Hz, H
a
), 7.53 (s, 1H, H20), 7.64 (dd, 1H, J ¼ 8.0/1.0 Hz,
H60), 7.97 (d, 1H, J ¼ 16.0 Hz, H
b d 56.07 (OCH3),
). 13C NMR (CDCl3)
60.92 (OCH3), 61.39 (OCH3), 101.78 (eOCH2Oe), 107.56 (C5), 107.84
(C20),108.45 (C50),121.00 (C6),122.09 (C1),123.88 (C60),124.49 (C
a
),
133.32 (C10), 139.58 (C
b
), 142.48 (C3), 148.19 (C30), 151.45 (C2),
2.2. Antiproliferative assays
153.75 (C40), 156.67 (C4), 188.65 (C]O). IR nmax/cmꢁ1 1652 (C]O),
1583 (C]C), 1249, 1041 (CeO), 3079, 2976, 2942, 2901, 2837, 1492,
1484, 1464, 1446, 1413, 1329, 1301, 1282, 1115, 1095, 988, 942, 916,
815, 697, 506 (Ar) (KBr). Anal. Calcd for C19H18O6: C 66.66, H 5.30.
Found: C 66.91, H 5.27. Yield: 94%.
2.2.1. L-1210 mouse leukemia cell assays
2.2.1.1. Cell culture. Murine lymphoblastic leukemia (L1210) cells
were obtained from American Type Culture Cell (ATCC). The cells
were cultured in RPMI (Roswell Park Memorial Institute medium)
supplemented with 10% heat-inactivated fetal bovine serum (FBS),
2.1.2.3. 3a e (2E,Z)-1-(30,40,50-trimethoxyphenyl)-3-(4-
dimethylaminophenyl)-2-propen-1-one. Golden yellow solid, m.p.
100 U/mL penicillin, 100 mg/mL streptomycin and 10 mM HEPES.
The cell culture was maintained at 37 ꢂC in a 5% CO2 humidified
atmosphere and pH 7.4. Every 2e3 days, cells were passaged by
removing 90% of the supernatant and replacing it with fresh me-
dium. In all experiments, viable cells were checked in the beginning
of the experiment by the Trypan Blue exclusion test.
148e149 ꢂC; 1H NMR (CDCl3)
d
2.98 (s, 3H, Z-CH3), 3.06 (s, 3H, E-
CH3), 3.88 (s, 6H, Z-m-OCH3), 33.90 (s, 3H, Z-p-OCH3), 3.93 (s, 3H, E-
p-OCH3), 3.95 (s, 6H, E-m-OCH3), 6.44 (d,1H, J ¼ 12.0 Hz, Z-H ), 6.58
(d, 1H, J ¼ 8.0 Hz, Z-H3, Z-H5), 6.71 (d, 1H, J ¼ 8.0 Hz, E-H3, E-H5),
a
6.90 (d, 1H, J ¼ 12.0 Hz, Z-H
b
), 7.19 (2, 2H, Z-H20, Z-H60), 7.27 (2, 2H,
E-H20, E-H60), 7.32 (d, 1H, J ¼ 16.0 Hz, E-H
a
), 7.56 (m, 2H, E-H2, E-
2.2.1.2. Cytotoxicity. Cytotoxicity was evaluated with the MTT ((3-
(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay
[37]. 1 ꢃ 105 cells/well were incubated for 24 h in triplicate with
compounds (solubilized in a maximum of 1% DMSO), at different
H6), 7.5e (m, 2H, Z-H2, Z-H6), 7.80 (d, 1H, J ¼ 16.0 Hz, E-H
b
). 13C
NMR (CDCl3)
(C20, C60), 112.04 (C3, C5), 116.77 (C1), 122.84 (C
134.71 (C10), 146.04 (C
d
40.36 (CH3), 56.57 (m-OCH3), 60.80 (p-OCH3), 106.06
), 130.66 (C2, C6),
), 141.20 (C40), 152.29 (C4), 153.27 (C30, C50),
a
b
concentrations (1e100
at 37 ꢂC, cells were washed with fresh culture medium, and 10
MTT (5 mg/mL) were added followed by incubation at 37 ꢂC for 2 h.
The precipitated formazan was dissolved in 100 L of DMSO, and the
mM) in 96-well microplates. After incubation
189.70 (C]O); in the RMN 13C was possible observe only the E-
isomer. IR nmax/cmꢁ1 1650 (C]O), 1563 (C]C), 1228, 1006 (CeO),
1153 (CeN), 2942, 2845, 1613, 1549, 1524, 1506, 1432, 1413, 1377,
1347, 1302, 1191, 1126, 1073, 984, 817, 669 (Ar) (KBr). Anal. Calcd for
mL of
m
absorbance was measured at 540 nm using a micro-well system
reader. The IC50 values were calculated from a four parameter logistic
equation. The chalcones were dissolved in DMSO, and in order to
verify if the solvent itself could affect the cells, in all experiments,
control curves without chalcones and in the presence of the cells and
thesolventwerecarriedoutinparallel.Thecontrolswithsolventwere
not statistically different from control cells alone (data not shown).
C
20H23NO4: C 70.36, H 6.79, N 4.10. Found: C 70.05, H 6.83, N 4.08.
Yield: 45%.
2.1.2.4. 3p e (2E)-1-(30,40,50-trimethoxyphenyl)-3-(5-methylfuran-2-
yl)-2-propen-1-one. Dark yellow solid, m.p. 89e90 ꢂC; 1H NMR
(CDCl3)
d 2.41 (s, 3H, CH3), 3.94 (s, 3H, p-OCH3), 3.95 (s, 6H, m-
OCH3), 6.14 (d, 1H, J ¼ 4.0 Hz, H4), 6.44 (d, 1H, J ¼ 4.0 Hz, H5), 7.29
(2, 2H, H20, H60), 7.32 (d, 1H, J ¼ 16.0 Hz, H
a
), 7.55 (d, 1H, J ¼ 16.0 Hz,
2.2.2. Cytotoxicity in tumorigenic and non-tumorigenic cell lines
2.2.2.1. Cell culture. REH and JURKAT human acute lymphoblastic
leukemia cell lines of precursor B-cell and T-cell immunopheno-
types, respectively, were cultured in RPMI-1640 medium supple-
mented with 10% FBS (RPMI-10), 100 U/mL penicillin and 100 pg/
mL streptomycin. Cultures were maintained at 37 ꢂC in a 5% CO2
humidified atmosphere. NIH3T3 and HUVEC were cultured under
the same conditions in RPMI-1640 and DMEM, respectively. Normal
Hb d 14.30 (CH3), 56.66 (m-OCH3), 61.17 (p-
). 13C NMR (CDCl3)
OCH3), 106.17 (C20, C60), 109.64 (C4), 117.23 (C5), 118.67 (C
a), 131.07
(Cb
), 133.96 (C10), 142.48 (C40), 150.65 (C1), 153.32 (C30, C50), 156.20
(C3), 188.85 (C]O). IR nmax/cmꢁ1 1653 (C]O), 1565 (C]C), 1227,
1012 (CeO), 2941, 2839, 1606, 1582, 1503, 1414, 1335, 1158, 1126,
796, 698, 625 (Ar) (KBr). Anal. Calcd for C17H18O5: C 67.54, H 6.00.
Found: C 67.76, H 6.42. Yield: 78%.