3
70
Vol. 54, No. 3
5
ꢀ-Chloro-2ꢀ-hydroxy-4-methoxychalcone (7): Yield, 38.0%. mp 188—
13)
1
90 °C (in the literature, 191—192 °C).
ꢀ-Bromo-2ꢀ-hydroxy-4-methoxychalcone (8): Yield, 81.0%. mp 102—
5
16)
1
04 °C (in the literature, 104 °C).
ꢀ-Hydroxy-4,6ꢀ-dimethoxychalcone (9): Yield, 48.5%. mp 96—98 °C (in
2
13)
the literature, 98—100 °C).
ꢀ-Hydroxy-4-methoxy-4ꢀ,5ꢀ-dimethylchalcone (10): H-NMR (DMSO-
1
2
d ) d: 12.63 (1H, s), 7.88 (1H, d, Jꢁ15.3 Hz), 7.51 (1H, d, Jꢁ15.3 Hz), 7.66
6
(
1H, s), 6.95 (1H, s), 7.62 (2H, d, Jꢁ8.7 Hz), 6.93 (2H, d, Jꢁ8.7 Hz), 3.86
(
1
n
3H, s), 2.27 (3H, s), 2.25 (3H, s). Tlc, Rf 0.57 (5 : 3 hexane/EtOAc). mp
ꢂ
42—144 °C. MS m/z: 282 (M ). Yield: 67.6%. Brown needles. IR (KBr)
ꢃ1
cm : 3403, 1623.
max
4
,2ꢀ,5ꢀ-Trihydroxychalcone (11): Yield, 21.1%. mp 191—192 °C (in the
17)
literature, 191—192 °C).
ꢀ,4ꢀ-Dihydroxy-4-methoxychalcone (12): Yield, 24.1%. mp 188—190 °C
(
2
18)
in the literature, 186 °C).
2
ꢀ,3,4,4ꢀ-Tetrahydroxychalcone (13): Yield, 21.5%. mp 250—252 °C (in
19)
the literature, 251—252 °C).
,2ꢀ,4,4ꢀ-Tetrahydroxychalcone (14): Yield, 28.6%. mp 138—140 °C (in
2
20)
the literature, 140 °C).
ꢀ,4ꢀ-Dihydroxy-4-methylchalcone (15): Yield, 16.0%. mp 102—104 °C
2
21)
(
in the literature, 104—106 °C).
ꢀ-Chloro-2ꢀ,3-dihydroxychalcone (16): H-NMR (DMSO-d ) d: 12.86
1
5
6
(
1H, s), 8.28 (1H, d, Jꢁ2.7 Hz), 7.99 (1H, d, Jꢁ15.3 Hz), 7.86 (1H, d,
Jꢁ15.3 Hz), 8.63 (1H, s) 7.01 (1H, dd, Jꢁ2.1, 8.7 Hz), 7.2—7.4 (2H, m),
.53 (1H, dd, Jꢁ2.4, 9.0 Hz), 6.96 (1H, m). Tlc, Rf 0.41 (5 : 3
7
ꢂ
hexane/EtOAc). mp 149—150 °C. MS m/z: 274 (M ). Yield: 21.8%. Yellow
needles. IR (KBr) n cm : 3344, 1641.
ꢃ1
max
5
ꢀ-Chloro-2ꢀ-hydroxy-4-dimethylaminochalcone (17): Yield, 52.0%. mp
22)
132—134 °C (in the literature, 134—135 °C).
General Procedure for Synthesis of TZD Chalcone Derivatives
A
mixture of 2ꢀ-hydroxyacetophenone and 50% aqueous potassium hydroxide
solution in 95% ethanol was added to 5-(4-formylphenylmethylene)-2,4-thi-
azolidinedione. After stirring for 5 h at room temperature, the reaction was
quenched with distilled water. The solution was acidified with 1 N HCl and
diluted with ethyl acetate. The organic phase was washed with brine, dried
over anhydrous magnesium sulfate, and concentrated. The residue was puri-
fied on column chromatography (silica gel with 20% ethyl acetate in hexane)
to give a yellow powder.
Fig. 1. PPAR-g Transient Transfection Assay
Activity was determined using transient transfection assays. Luciferase activity was
determined and plotted as fold activation relative to untreated cells. PGJ2: 15-deoxy-
1
2,14
-PGJ2.
D
5
-{4-[3-(2ꢀ-Hydroxyphenyl)-3-oxo-propenyl]-benzylidene}-thiazolidine-
1
a more detailed structure–activity relationship study of the 2,4-dione (18): H-NMR (DMSO-d ) d: 12.8 (1H, s), 8.18 (1H, d,
6
present series of chalconylidene-TZD derivatives are in Jꢁ15.6 Hz), 7.67 (1H, d, Jꢁ15.6 Hz), 8.01 (1H, d, Jꢁ8.1 Hz), 7.02 (1H, d,
Jꢁ8.1 Hz), 7.6—7.7 (1H, m), 6.99 (1H, d, Jꢁ8.1 Hz), 7.71 (2H, d,
progress.
1
3
Jꢁ8.4 Hz), 7.36 (2H, d, Jꢁ8.4 Hz), 6.98 (1H, s). C-NMR (DMSO-d ) d:
27.0, 162.0, 117.9, 136.6, 121.1, 130.0, 193.6, 123.5, 147.8, 135.8, 131.1,
30.5, 135.9, 143.8, 119.4, 168.8, 169.5. Tlc, Rf 0.36 (5 : 3 MC/MeOH). mp
86—189 °C. MS m/z: 351 (M ). Yield: 36.0%. Yellow powder. IR (KBr)
6
1
1
1
n
Experimental
1
13
Chemistry H- (300 MHz) and C-NMR (75 MHz) spectra were ob-
tained with a Varian Gemini-2000 spectometer. Chemical shifts (d) are re-
ported in ppm relative to tetramethylenesilane as internal standard. Melting
points were determined on a Mitamura-Riken melting point apparatus
ꢂ
ꢃ1
cm : 3287, 1651.
max
5
-{4-[3-(2ꢀ-Hydroxy-5ꢀ-methylphenyl)-3-oxo-propenyl]-benzylidene}-
1
thiazolidine-2,4-dione (19): H-NMR (DMSO-d ) d: 13.1 (1H, s), 8.08 (1H,
d, Jꢁ15.3 Hz), 7.81 (1H, d, Jꢁ15.3 Hz), 7.67 (1H, d, Jꢁ9.0 Hz), 7.54 (1H, d,
Jꢁ9.0 Hz), 7.46 (1H, d, Jꢁ9.0 Hz), 7.62 (2H, d, Jꢁ9.0 Hz), 6.94 (2H, d,
Jꢁ9.0 Hz), 6.88 (1H, s), 3.81 (3H s). C-NMR (DMSO-d ) d: 124.2, 152.0,
18.9, 121.0, 156.1, 113.5, 193.1, 123.7, 143.5, 131.5, 130.5, 130.0, 134.8,
37.8, 168.8, 169.5, 55.9. Tlc, Rf 0.46 (5 : 3 MC/MeOH). mp 123—126 °C;
6
(Japan) and are uncorrected. IR spectra were recorded on a JASCO FT/IR-
5300 spectrometer, and UV spectra were obtained with a Hitachi U-3210
UV–VIS spectrophotometer. Mass spectra were recorded on a Hewlett
Packard model 5989 B GC/MS system and VG70-VSEQ (VG analytical,
U.K.).
General Procedure for Synthesis of Chalcone Derivatives To a mix-
ture of 2ꢀ-hydroxyacetophenone (3.70 mmol) and potassium hydroxide
13
6
1
1
ꢂ
ꢃ1
MS m/z: 381 (M ). Yield: 35.0%. Yellow powder. IR (KBr) n cm
:
max
3
269, 1641.
-{4-[3-(2ꢀ-Hydroxy-5ꢀ-methoxyphenyl)-3-oxo-propenyl]-benzylidene}-
(
(
18.5 mmol) in 20 ml of 95% ethanol was added 4-methoxybenzaldehyde
3.70 mmol). After stirring for 5 h at room temperature, the reaction was
5
1
thiazolidine-2,4-dione (20): H-NMR (DMSO-d ) d: 12.3 (1H, s), 8.08 (1H,
d, Jꢁ15.3 Hz), 7.83 (1H, d, Jꢁ15.3 Hz), 7.76 (1H, d, Jꢁ9.0 Hz), 7.47 (1H, d,
Jꢁ8.7 Hz), 7.33 (1H, d, Jꢁ8.7 Hz), 7.62 (2H, d, Jꢁ8.7 Hz), 7.01 (2H, d,
Jꢁ8.7 Hz), 6.90 (1H, s), 2.49 (3H, s). C-NMR (DMSO-d ) d: 123.2,
60.1, 117.6, 137.3, 130.3, 131.2, 193.5, 126.9, 143.5, 135.8, 128.2, 127.9,
36.0, 143.5, 120.5, 168.8, 169.4, 20.9. Tlc, Rf 0.56 (5 : 3 MC/MeOH). mp
54—157 °C; MS m/z: 365 (M ). Yield: 25.0%. Yellow powder. IR (KBr)
nmax cm : 3273, 1651.
In Vitro Biological Activity The PPAR-g ligand-binding activity of the
chalcones (3—17) and chalconylidene-TZD (18—20) was determined using
transient transfection assays. The CV-1 cells were plated in 96-well plates
at 1.2ꢄ10 cells/well and cultured until 70—80% confluency for 16 h. The
cells were transiently transfected with pCDM8-mPPARg, pcDNA3.1-
mRXRa, pPPRE (ꢄ3) tk-luc, and pcDNA3.1-lacZ by liposomal delivery
using a GenePORTER2 transfection kit (GTS). After 16-h incubation, the
cells were washed and changed to fresh complete medium with the indicated
6
quenched with distilled water. The resulting solution was acidified with 1 N
HCl and extracted with ethyl acetate. The organic phase was washed with
brine, dried over anhydrous magnesium sulfate, filtered, and concentrated.
The residue was purified on column chromatography (silica gel, eluted with
13
6
1
1
1
33% ethyl acetate in hexane). In the case of synthesis of compounds with a
2
ꢀ,4ꢀ- or 3,4-dihydroxyl group, the hydroxyl group of the component was
ꢂ
protected with methoxymethyl using chloro methylmethyl ether.
ꢀ-Hydroxy-4-methoxychalcone (3): Yield, 33.0%. mp 87—90 °C (in the
ꢃ1
2
13)
literature, 95 °C).
ꢀ-Hydroxy-4,5ꢀ-dimethoxychalcone (4): Yield, 43.0%. mp 85—86 °C (in
2
23)
14)
the literature, 87—89 °C).
ꢀ,5ꢀ-Dihydroxy-4-methoxychalcone (5): Yield, 33.0%. mp 176—178 °C
4
2
1
5)
(in the literature, 178—80 °C).
2
ꢀ-Hydroxy-4-methoxy-5ꢀ-methylchalcone (6): Yield, 62.1%. mp 94—
16)
96 °C (in the literature, 92—94 °C).