Journal of Chemistry
3
2
H, H-3, H-5, Jm � 1.2, Jo � 8.8 Hz), 7.55–7.57 (m, 2H, H-4′,
of 4-hydroxybenzaldehyde (8), vanillin (9), or isovanillin (10))
H-7′), 7.57 (dd, 2H, H-2, H-6, Jm � 1.2, Jo � 8.8 Hz), 7.67 (s,
and K CO (0.51 mmol, 2 equiv.) in 5 mL of acetonitrile was
2
3
13
°
1
H, �C-H), and 12.68 (s, 1H, N-H) ppm. C NMR
heated at 50 C for 30 min. After that, 2-(chloromethyl)-1H-
benzimidazole (4) (0.275 mmol, 1.1 equiv.) in acetonitrile was
(
100 MHz, DMSO-d ) δ: 64.4 (OCH ), 116.1 (C-3, C-5, C-4′,
6
2
C-7′), 122.4 (C-5′, C-6′), 123.1 (�C-), 127.1 (C-1), 130.7
added drop by drop. ꢀe reaction was heated for 2–9 h.
Acetonitrile was removed using a rotavapor. 10 mL of ice-water
was added and stirred for 10 min. ꢀe emulsion was extracted
with EtAcO (3 ×15 mL), and the organic layer was evaporated
to give a solid, which was recrystallizated from suitable solvent.
(
�C-H), 132.3 (C-2, C-6), 138.6 (C-3a′, C-7a′), 150.1 (C-2′),
1
59.7 (C-4), 168.1 (N-C�O), 171.1 (S-C�O) ppm; MS/EI: m/
+
z (% int. rel). 351 (M , 1%), and 259 (100%). Anal. calcd. for
C H N O S: C 61.53, H 3.73, N 11.96, S 9.13; found C
1
8
13 3 3
6
1.50, H 3.68, N 11.87, S 9.19.
2
.4.1. 4-(1H-Benzimidazol-2-ylmethoxy)benzaldehyde (5).
1
2
.2.2. (5Z)-5-[4-(1H-Benzimidazol-2-ylmethoxy)-3-methox-
Brown powder, ethanol, mp 179°C–181.9°C; yield 95%. H
ybenzylidene]-1,3-thiazolidine-2,4-dione (2). Yellow pow-
NMR (400 MHz, DMSO-d ) δ: 5.45 (s, 2H, OCH ), 7.17–
6
2
1
°
der, mp 245.8–248.7 C; yield 85.5%. H NMR (400 MHz,
DMSO-d ) δ: 3.80 (s, 3H, OCH ), 5.36 (s, 2H, OCH ), 7.14
7.20 (m, 2H, H-5′, H-6′), 7.29 (dd, 2H, H-3, H-5, Jm � 1.2,
Jo � 8.8 Hz), 7.54–7.56 (m, 2H, H-4′, H-7′), 7.88 (dd, 2H,
H-2, H-6, Jm � 1.2, Jo � 8.8 Hz), and 9.87 (s, 1H, CHO) ppm.
6
3
2
(
d, 2H, H-5′, H-6′ Jo � 8.4 Hz), 7.18–7.20 (m, 2H, H-2, H-6),
+
7
7
.30 (d, 1H, H-5, Jo � 8.8 Hz), 7.55–7.57 (m, 2H, H-4′, H-7′),
MS/EI: m/z (% int. rel). 252 (M , 24%) and 131 (100%).
13
.71 (s, 1H, �C-H), and 12.68 (s, 1H, N-H) ppm. C NMR
(
100 MHz, DMSO-d ) δ: 55.7 (OCH ), 64.7 (OCH ), 113.9
6
3
2
2
.4.2. 4-(1H-Benzimidazol-2-ylmethoxy)-3-methoxybenzaldehyde
(
C-5), 114.1 (C-2), 121.17 (�C-), 123.8 (�C-H), 127.1 (C-6),
1
°
(
6). Brown powder, ethanol, mp 71.7–74.7 ; yield 48.7%. H
1
3
28.6 (C-4′, C-7′), 129.1 (C-5′, C-6′), 132.1 (C-1), 138.6 (C-
a′, C-7a′), 149.8 (C-4), 149.8 (C-3), 149.5 (C-2′), 168.1 (N-
NMR (400 MHz, DMSO-d ) δ: 3.82 (s, 3H, OCH ), 5.41 (s,
6
3
2
H, OCH ), 7.16–7.18 (m, 2H, H-5′, H-6′), 7.37 (d, 1H, H-5,
2
C�O), and 168.6 (S-C�O) ppm; MS/EI: m/z (% int. rel). 381
+
Jo � 8.8 Hz), 7.41 (d, 1H, H-2, Jm � 2.1 Hz), 7.53 (dd, 1H, H-6,
(
M , 1%), 265 (80%), and 180 (100%). Anal. calcd. for
Jm � 2.1, Jo � 8.8 Hz), 7.53–7.55 (m, 2H, H-4′, H-7′), and 9.83
C H N O S : C 59.83, H 3.96, N 11.02, S 8.41; found C
1
9
15 3 4
+
(
s, 1H, CHO) ppm. MS/EI: m/z (% int. rel) 282 (M , 1%) and
5
9.90, H 3.89, N 11.00, S 8.45.
1
31 (100%).
2
.2.3. (5Z)-5-[3-(1H-Benzimidazol-2-ylmethoxy)-4-methox-
2
.4.3. 3-(1H-Benzimidazol-2-ylmethoxy)-4-methoxybenzaldehyde
ybenzylidene]-1,3-thiazolidine-2,4-dione (3). Yellow pow-
1
1
°
°
(7). Brown powder, ethanol, mp 80.9 C (dec); yield 59%. H
der, mp 246.9 C (dec); yield 31%. H NMR (400 MHz,
DMSO-d ) δ: 3.85 (s, 3H, OCH ), 5.34 (s, 2H, OCH ), 7.17 (s,
NMR (400 MHz, DMSO-d ) δ: 3.89 (s, 3H, OCH ), 5.37 (s,
2
6
3
6
3
2
H, OCH ), 7.13 (d, 1H, H-5, Jo � 8.7 Hz), 7.19 (m, 2H, H-4′,
1
H, H-5, Jo � 8.6 Hz), 7.18–7.20 (m, 2H, H-5′, H-6′), 7.23
2
H-7′), 7.23 (d, 1H, H-6 Jo � 8.7 Hz), 7.37 (s, 1H, H-2), and
(
dd, 1H, H-6, Jm � 1.02, Jo � 8.6 Hz), 7.37 (d, 1H, H-2,
+
9
.82 (s, 1H, CHO) ppm. MS/EI: m/z (% int. rel) 282 (M , 2%)
Jm � 1.02 Hz), 7.56–7.58 (m, 2H, H-4′, H-7′), 7.69 (s, 1H,
13
and 131 (100%).
�
C-H), and 12.68 (s, 1H, N-H) ppm. C NMR (100 MHz,
DMSO-d ) δ: 55.7 (OCH ), 64.4 (OCH ), 112.5 (C-5), 115.2
6
3
2
(
C-2), 122.1 (�C-), 124.6 (C-6), 125.7 (�C-H), 128.5 (C-4′,
2
2
.5. Biological Activity
C-7′), 129.2 (C-5′, C-6′), 131.3 (C-1), 138.6 (C-3a′, C-7a′),
1
1
47.5 (C-3), 149.5 (C-4), 150.1 (C-2′), 168.4 (N-C�O), and
.5.1. PPARc and GLUT-4 Determination. Assays were
+
68.8 (S-C�O) ppm; MS/EI: m/z (% int. rel). 381 (M , 1%),
performed as described elsewhere [3, 12, 13]. Briefly, 3T3-
L1 murine fibroblasts were cultured, and cell viability was
measured using the MTT assay at three increasing con-
centrations (1, 10, and 100 μM) of hybrids 1–3. Confluent
cultures of fibroblasts were differentiated to the adipocyte
phenotype for 48 h employing 0.5 mM of 3-isobutyl-1-
methylxanthine, 0.25 μM of dexamethasone acetate, and
anal. calcd. for C H N O S : C 59.83, H 3.96, N 11.02, S
1
9
15 3 4
8
.41; found C 59.76, H 3.89, N 11.00, S 8.36.
2
1
.3. Preparation of 2-(Chloromethyl)-1H-benzimidazole (4).
,2-Phenylenediamine (1 g, 0. 8 mmol), 1.3 equivalents of
°
chloroacetic acid, and 10 mL of 1 M HCl were heated at 90 C
in a nitrogen atmosphere for 10 h. ꢀe cooled mixture was
0
.8 μM of murine insulin, enriched by additional insulin
charge. After 8 to 10 days, the cells acquire the mature
adipocyte phenotype and were treated for 24 h to observe
the effects of heterocycles 1–3 on PPARc and GLUT-4
mRNA expression. Total mRNA was isolated from adi-
pocytes, and 2 μg was reverse transcribed. ꢀe cDNA was
amplified using SYBR Green Master Mix (Roche) con-
taining 0.5 mM of customized primers for PPARc (Gen-
Bank accession no.: NM 011146.1) and GLUT-4 (GenBank
accession no.: NM009204.2). PCR was used for individually
sample calculating the threshold cycles (C ) and the ΔC
basified with saturated NaHCO solution, and the crude
3
product was extracted with AcOEt. Solvent was removed
°
and the solids filtrated. Yellow powder, mp 147.8–148.2 C
1
°
°
(
Lit. 142 C–144 C); yield 53.2%. H NMR (400 MHz,
DMSO-d ) δ: 4.90 (s, 2H, ClCH ), 7.18–7.20 (m, 2H, H-5′,
6
2
H-6′), and 7.54–7.55 (m, 2H, H-4′, H-7′) ppm. MS/EI: m/z
+
(
% int. rel). 165 (M , 45%), 131 (100%), and 180 (100%).
2
.4. General Preparation of Precursors 5–7. A solution of
t
t
generally recognized as safe hydroxybenzaldehydes (0.25 mmol
values. ꢀe amount of mRNA for each gene was normalized