Synthesis and biological evaluation of sulfonylurea and thiourea derivatives substituted with benzenesulfonamide groups as potential hypoglycemic agents

Article abstract of DOI:10.1016/j.bmcl.2009.01.082

A novel class of sulfonylurea and thiourea derivatives substituted with benzenesulfonamide groups were designed and synthesized. The target compounds were assayed for the effects on the insulin release of isolated rat pancreatic islets and the glucose transport in adipocytes of rats. Some of them exhibited high potency. Compound 10 also had potent antiplatelet activity and showed an excellent property to protect collagen-epinephrine-induced mice mortality as well as plasma glucose-lowering activity in vivo. The preliminary pharmacological profile of compound 10 showed that it might be useful in the treatment of diabetics with cardiovascular and nephropathy complications.

Full text of DOI:10.1016/j.bmcl.2009.01.082

Bioorganic & Medicinal Chemistry Letters 19 (2009) 1740–1744
Contents lists available at ScienceDirect
Bioorganic & Medicinal Chemistry Letters
journal homepage: www.elsevier.com/locate/bmcl
Synthesis and biological evaluation of sulfonylurea and thiourea derivatives
substituted with benzenesulfonamide groups as potential hypoglycemic agents
a,
c
Hui-bin Zhang a, Ya-an Zhang a, Guan-zhong Wu b, Jin-pei Zhou a, , Wen-long Huang , Xiao-wen Hu
*
*
a Center of Drug Discovery, China Pharmaceutical University, 24 Tongjiaxiang, Nanjing, Jiangsu 210009, China
b Department of Pharmacology, China Pharmaceutical University, 24 Tongjiaxiang, Nanjing 210009, China
c Department of Math and Computer, Nanjing Medical University, 140 Hanzhonglu, Nanjing 210029, China
a r t i c l e i n f o
a b s t r a c t
Article history:
A novel class of sulfonylurea and thiourea derivatives substituted with benzenesulfonamide groups were
designed and synthesized. The target compounds were assayed for the effects on the insulin release of
isolated rat pancreatic islets and the glucose transport in adipocytes of rats. Some of them exhibited high
potency. Compound 10 also had potent antiplatelet activity and showed an excellent property to protect
collagen–epinephrine-induced mice mortality as well as plasma glucose-lowering activity in vivo. The
preliminary pharmacological profile of compound 10 showed that it might be useful in the treatment
of diabetics with cardiovascular and nephropathy complications.
Received 27 July 2008
Revised 8 January 2009
Accepted 24 January 2009
Available online 29 January 2009
Keywords:
Sulfonylurea derivatives
Hypoglycemic activity
Diabetic complications
Synthesis
Ó 2009 Elsevier Ltd. All rights reserved.
Patients with diabetes mellitus are at high risk for developing
long-term complications including neuropathy, nephropathy, reti-
nopathy, and cardiovascular disease.1 Atherosclerotic macrovascu-
lar disease accounts for the majority of morbidity and mortality
associated with diabetes.2 Diabetic nephropathy, as a serious
chronic diabetic microvascular complication, has become a single
most important cause of end-stage renal disease.3 So a more com-
prehensive approach aimed at reducing cardiovascular and neph-
ropathic complications as well as controlling glucose excursion
may prove more beneficial to diabetic patients.
betic rats suggested that endogenous TXA2 receptor agonists stim-
ulate the development of diabetic nephropathy.13
There were four main pathways proposed for the mechanisms
of hyperglycemia-associated tissue damage: the polyol pathway,
the hexosamine pathway, protein kinase C activation, and the for-
mation of advanced glycation end-products (AGE). But there was
no apparent common element linking these mechanisms to each
other. Clinical trials of inhibitors of these pathways in patients
were also disappointing. A unifying mechanism proposed by Mi-
chael Brownlee14 is the overproduction of superoxide by the mito-
chondrial electron transport chain. Increased production of ROS
and oxidative stress is assumed to play a central role in the path-
ogenesis of diabetes complications.
S18886 (terutroban), a benzenesulfonamide-substituted tetra-
hydronaphthalene derivative, is an orally active, high potent, selec-
tive TPr antagonist.15 It exerts potent antiplatelet and
antivasoconstrictory effects via the TPr and antagonizes the actions
of TXA2, as well as of other arachidonic acid products.16 The anti-
atherosclerotic effect of S18886 has also been demonstrated in rab-
bits.17,18 Several articles published recently had reported that
S18886 can significantly inhibit atherogenesis in the non-diabetic
and diabetic ApoEÀ/À mice.19,20 It has also been reported that
S18886 had the nephroprotective action in the diabetic ApoEÀ/À
mice.20 ApoEÀ/À mice are animals genetically deficient in TPr, so
the mechanism of antiatherosclerotic and nephroprotective action
of S18886 could be independent of platelet-derived TXA2.19 Further
studies indicated that S18886 can enhance the antioxidative de-
fense.21 The effect of inhibiting TPr can be attributed to a novel role
The increased atherosclerotic risk in diabetic patients4 was
associated with many factors, mainly including hyperglycemia,
insulin resistance, pro-inflammatory and prothrombotic status.5
In particular, the prothrombotic status is related to endothelial
dysfunction, impaired fibrinolysis, increased coagulation factors,
and importantly, increased platelet reactivity.6,7 Antiplatelet drugs
may have therapeutic potential in prevention or treatment for dia-
betic cardiovascular complications. On the other hand, elevated
glucose in diabetes has long been recognized to increase levels of
arachidonic acid metabolites, many of which stimulate TXA2–
PGH2 receptor (TPr).8,9 The TXA2/PGI2 ratio of diabetics was signif-
icantly higher than that of healthy volunteers.10 Earlier studies
with TXA2 receptor antagonists Bay U340511 and S-145212in dia-
* Corresponding authors. Tel.: +86 25 83471532; fax: +86 25 83271480.
E-mail addresses: jpzhou668@163.com (J. Zhou), ydhuangwenlong@126.com
(W. Huang).
0960-894X/$ - see front matter Ó 2009 Elsevier Ltd. All rights reserved.
doi:10.1016/j.bmcl.2009.01.082

H. Zhang et al. / Bioorg. Med. Chem. Lett. 19 (2009) 1740–1744
1741
of TPr in mediating oxidant stress in the kidney. TP receptors play
an important role in atherogenesis and nephropathy which are clo-
sely related to diabetes.
chloride 3, which was converted into the corresponding benzene-
sulfonamide 4 by stirring overnight in concentrated ammonia
water. Hydrolysis of compound 4 with sodium hydroxide solution
afforded the key intermediate 4-(2-aminoethyl) benzenesulfon-
amide 5. The benzenesulfonamide 5 reacted with substituted ben-
zenesulfonyl chloride to form 4-(2-substituted benzenesulfonyl
aminoethyl) benzenesulfonamide 6. The target compounds 7–25
were prepared by the reaction of compound 6 with isocyanate or
isothiocyanate. It was reported that the reaction between com-
pound 6 and isocyanate should be under the condition of strong
base in anhydrous solvents.24 We treated compound 6 with anhy-
drous potassium carbonate in refluxing anhydrous acetone for 6 h,
to form potassium salt of compound 6, then added the solution of
isocyanate in anhydrous acetone dropwise to the potassium salt
above, refluxed for another 6 h, we got the target compounds in
reasonable yields (56.5–92.4%).25
Although the source of ROS is controversial22 and the most ele-
ments in oxidant stress remains uncertain, the potent therapeutic
potential of TPr antagonist in non-diabetic and diabetic atheroscle-
rosis and early diabetic nephropathy is conceivable. With its
important role in the oxidant stress, TPr antagonism can enhances
the antioxidative defense, thereby effectively reducing ROS-in-
duced downstream tissue damage in the development of diabetic
cardiovascular and nephropathy complications.
Doltroban (BM 13,505) and sulotraban (BM 13,177) are two
early reported platelet TXA2 receptor antagonists23 The benzene-
sulfonamide is the common moiety in their structures and has
been proven to be the pharmacophore of the TPr antagonists.
S18886 also has the similar structural backbone with doltroban
and sulotraban (as seen in Figure 1).
The sulfonylureas, for example, glimepiride and glibenclamide,
were the most widely used oral antidiabetic drugs in clinic. Glim-
epiride also has blood platelet aggregation inhibiting property. So
we hypothesized that TPr antagonistic and hypoglycemic activities
may have some inherent relevancy with compounds with ben-
zenesulfonamide and sulfonylurea structures.
The immediate medicinal chemistry strategy became the
replacement of the benzoyl or heterocyclic formacyl groups, which
are commonly present in sulfonylurea drugs, with benzenesulfon-
amide group which is the functional group of the blood platelet
TXA2 receptor antagonists. We also changed the urea group with
its bioisosterist thiourea to check the change of activity. From these
results, we designed a fused skeleton of doltroban and glimepiride
structures (Scheme 1) with the expectation that the target com-
pounds will show effects on diabetic complications such as
nephropathy and atherosclerosis as well as hypoglycemic activity.
We reported here the synthesis of new sulfonylurea and thiourea
derivatives substituted with benzenesulfonamide groups and their
in vitro and in vivo pharmacological characterization of hypoglyce-
mic and antithrombotic properties related to TPr antagonism.
The target compounds were prepared as presented in Scheme 1.
The synthesis started with the acetylation of phenylethylamine 1
with acetic anhydride in triethylamine under room temperature,
to form N-acetylphenylethylamine 2. Chlorosulfonation of 2 with
chlorosulfonyl acid gave 4-(2-acetylaminoethyl) benzenesulfonyl
The target compounds described in Scheme 1 were tested for
in vitro insulin secretion activity with isolated pancreatic cells.
The method was described in Ref. 26. The results were showed in
Table 1.
The test demonstrated that insulin release of pancreatic cells in-
creased by 5.49 times after glucose treatment and by 2.03 times
after glimepiride (100 mol/L) compared with basal. The sulfourea
derivatives with aryl substituents (17–25) showed very little activ-
ity. The urea compounds with phenylsulfonylamide groups (7–16)
retained insulin release activity. But the amino- or methoxyl-
substituted phenylsulfonylamides (7, 8, and 11) were less active,
showed moderate activity. The halo-substituted compounds (9,
10, 15, and 16) exhibited high potent. The compounds 10 and 9
with bromo or chloro atoms on the para-position of phenylsulfo-
nylamide have the highest insulin release property, increased by
3.64 and 3.05 times, respectively, and higher than that of glimepi-
ride at the same concentration (2.03 times).
Insulin can stimulate the glucose transport in adipocytes.27 Sul-
fonylurea drugs have the effect to increase the glucose transport.
The target compounds were also tested for the effect on glucose
transport in adipocytes of rats in vitro (without/with 1.2 nM insu-
lin) following the method described.28 The results were showed in
Table 2.
Glimepiride has the highest activity to increase the glucose
transport in all sulfonylurea drugs. Treated with glimepiride
(100
lg/ml), glucose absorption of adipocytes increased by 2.5
H3C
N CONHCH2CH2
SO2NHCONH
Glimepiride
SO2NHCONH
10
C2H5
O
hypoglycemic moirty
Br
SO2NHCH2CH2
Cl
SO2NHCH2CH2
CH2COOH
Doltroban
antagonistic
moicty
SO2NHCH2CH2
OCH2COOH
Sulotraban
Cl
SO2NH
H
CH2COOH
S18886
Figure 1. The structures of Glimepiride, compound 10, Doltroban, Sulotraban, and S1886.

1742
H. Zhang et al. / Bioorg. Med. Chem. Lett. 19 (2009) 1740–1744
a
b
CH2CH2NH2
CH2CH2NHCOCH3
CH3CONHCH2CH2
SO2Cl
3
1
2
c
d
e
NH2CH2CH2
SO2NH2
CH3CONHCH2CH2
SO2Cl
5
4
O
SO2NHCNH
f
R1
R2
SO2NH2CH2CH2
SO2NH2
R1
R2
SO2NH2CH2CH2
R3
6
7-16
g
7: R1=NHCOCH3; R2=H; R3=CH3
8: R1=NHCOCH3; R2=CH3; R3=CH3
9: R1=Cl; R2=H; R3=CH3
S
R1
SO2NH2CH2CH2
SO2NHCNH
R2
10: R1=Br; R2=H; R3=CH3
11: R1=NHCOCH3; R2=OCH3; R3=CH3
12: R1=H; R2=H; R3=H
17-25
17: R1=H; R2=Cl
22: R1=Br; R2=OCH3
23: R1=CH3; R2=Cl
18: R1=H; R2=CH3
19: R1=H; R2=OCH3
20: R1=Cl; R2=Cl
13: R1=CH3; R2=H; R3=H
24: R1=CH3; R2=CH3
25: R1=CH3; R2=OCH3
14: R1=H; R2=H; R3=CH3
15: R1=Cl; R2=H; R3=H
21: R1=Cl; R2=OCH3
16: R1=Br; R2=H; R3=H
Scheme 1. Reagents and conditions: (a) (CH3CO)2O, Et3N, rt; (b) ClSO3H, 0 °C, 6 h; (c) NH3ÁH2O, rt, 12 h, 89.9%; (d) NaOH, H2O, reflux, 3 h, 78%; (e) ArSO2Cl, K2CO3, DCM–H2O;
(f) K2CO3, anhydrous acetone, reflux 6 h then add isocyanate to reflux another 6 h, 56.5–92.4%; (g) K2CO3, anhydrous acetone, isothiocyanate as (f).
Table 1
fasted C57BL/6J mice were used to assess the ability of the cho-
Effects of compounds on insulin release of cells isolated from pancreatic islets of rats
sen compound to lower plasma glucose level.29The results were
Compound
Insulin secretiona (uIU/ml) Concn. (lmol/L)
shown in Figure 2. Three doses (20, 7, and 2.5 mg/kg) of com-
pound 10 obviously reduced the blood glucose level of the mice
and showed the remarkable dose relevance. At the dose of
20 mg/kg, the glucose-lowering property was about to 50%, the
duration is more than 6 h. The potency was similar to that of
glimepiride.
The compound 10 was evaluated for its ability to inhibit platelet
aggregation of human platelet-rich plasma (PRP) induced by ara-
chidonic acid or adenosine diphosphate with the method de-
scribed.30 The concentrations that caused 50% inhibition of
platelet aggregation (IC50) was obtained by regression analysis of
the concentration versus inhibition of aggregation curve. The re-
sults were shown in Table 3.
1
10
100
Control
Vehicle (0.25%
DMSO)
Glucose
(25 mmol/L)
Glimepiride
7
8
25.12 1.92
23.65 1.89
129.85 11.15c
48.14 3.13c
30.18 2.94b 36.67 3.24c 28.52 2.27a
23.57 1.45
27.67 1.21a 37.32 2.22b
9
38.15 2.44c 41.29 3.80c 72.23 5.38c
40.79 3.08c 65.21 4.26c 86.08 6.45c
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
24.57 2.14
22.86 2.25
22.45 2.04
24.47 1.33
28.27 1.03a 31.29 2.15b
22.70 1.31 25.93 1.75
24.83 1.64
32.15 2.44b 41.29 3.80c 52.23 5.38c
33.95 2.04b 41.18 3.16c 55.03 5.31c
32.09 2.68b 45.01 4.07c 65.98 6.25c
28.94 2.04a 32.10 2.16b 26.06 1.60
27.99 1.35a 27.71 1.84a 24.97 1.79
IC50 of compound 10 and Asp to AA were 0.21 and 0.35
lmol/L,
and IC50 to ADP were 3.00 and 9.34 mol/L, respectively. The re-
l
sults showed that compound 10 had potent antiplatelet activity
and was higher than Asp.
Protective effect of compound 10 against collagen–epinephrine-
induced mice mortality was performed on Chinese Kunming mice
following the method described.31 The effects of compound 10 or
Asp were evaluated by measuring the incidence of death within
15 min of injection. Results are expressed as percent survival. Each
group was comprised of 11 mice.
22.86 2.53
24.98 1.23
23.42 2.18
26.07 2.36
26.09 2.13
26.17 2.21
22.25 2.01
20.87 1.91
23.69 1.06
29.46 2.29a 26.37 1.64
24.05 1.34
22.25 1.54
23.89 2.57
21.99 2.03
24.48 1.43
24.36 1.12
24.67 2.02
23.01 2.19
24.24 1.67
23.50 1.45
The results were shown in Table 4. Three groups (40, 20, and
10 mg/kg) of compound 10 obviously reduced mice mortality and
showed the remarkable dosage relevance. At the dose of 40 mg/
kg of compound 10, the percentage of protection was 72.7% while
that of ASP (60 mg/kg) was 63.6%. So compound 10 showed
remarkable protective effect against collagen–epinephrine-in-
duced mice mortality.
Each value represents means SD of four determinations. aP < 0.05, bP < 0.01,
cP < 0.001 compared to Vehicle (0.25% DMSO). P < 0.05 was taken as the level of
significance.
times compared to basic absorption. Some of the compounds
tested showed relatively high activity. The compounds 10, 23, 11,
The preliminary pharmacokinetic properties of compound 10
was determined in rats (Table 5). It had a small volume of distribu-
tion (Vd = 0.78 0.36 L/kg), which suggested high plasma protein-
binding rate, similar to that of other sulfonylurea drugs (e.g., Glim-
epiride).32 The oral half-life was 1.77 0.21 h, and the oral bioavail-
ability was determined to be 52.96%. Compound 10 showed similar
pharmacokinetic properties with other sulfonylurea drugs.33
and 9 (100 lg/ml) increased glucose transport by 1.73, 1.59, 1.58,
and 1.57 times, respectively. Some compounds (8, 10, 16, 19, and
23) showed synergistic effect with insulin to increase the glucose
transport. The SAR was somewhat different to that of insulin re-
lease activity.
After determining the in vitro properties of compounds, com-
pound 10 was chosen to in vivo experiments. In the first, the

H. Zhang et al. / Bioorg. Med. Chem. Lett. 19 (2009) 1740–1744
1743
Table 2
Effects of compounds on glucose transport in adipocytes of rats in vitro (without/with insulin)
Compound
D
-1-3H-Glucose (dpm) Concn (
l
M, without/with 1.2 nM insulin)
1
10
100
Control
4501.2 278.44/
5379.33 227.19b
4427.1 179.05/
5477.12 136.44b
16239.2 1131.40c
Vehicle (0.25% DMSO)
Insulin (10 nM)
Glimepiride
5061.2 579.10a/
6365.67 378.57dg
4215.36 343.88/
5087.78 403.47
4302.19 328.31/
6123.02 202.43dh
5869.85 380.34a/
5942.68 226.19
4871.74 171.54a/
5996.43 229.52
5294.78 473.18a/
5587.26 307.42
7476.6 816.5c/
8593.72 382.70fh
5616.15 218.29b/
6200.35 159.650
4573.80 368.89/
6516.90 505.34d,h
6411.70 252.69b/
6358.77 467.25d
6512.65 288.59b/
7979.94 183.76e,h
6547.93 511.95b/
6341.17 470.32d
Not tested
11068.51 792.67c/
13255.41 916.73f,h
6736.82 545.69b/
6812.33 428.54e
5207.81 389.64a/
7244.87 322.08eh
6960.26 545.75b/
7013.28 327.48e
7674.84 220.06c/
8486.79 303.85fh
7037.78 409.57c/
7447.72 486.62e
7
8
9
10
11
12
13
4281.30 19.56/
5374.21 436.29
4174.59 394.56/
5533.15 346.28
Not tested
4458.36 256.10/
5297.11 401.64
14
15
4576.35 400.97/
5217.31 325.19
4421.72 151.54/
5523.89 425.52
5878.83 420.22a/
5933.89 296.52
4861.54 169.52a/
6002.46 245.55d
Not tested
6632.70 251.62b/
6521.12 362.28e
6556.61 284.52b/
7896.14 273.72eh
16
17
18
4377.95 266.81/
5319.50 354.27
4179.50 322.49/
5708.43 159.63
4587.26 156.24/
5179.30 441.02
4385.12 199.67/
5644.89 410.65
4118.78 411.45/
7555.71 316.32f,i
4432.15 355.65/
5436.58 284.56
Not tested
4026.52 387.24/
5234.61 358.10
6514.04 371.32b/
5512.62 454.58
4401.01 155.02/
5115.32 223.52
4381.35 369.15/
5377.15 246.51
4663.52 284.63/
5761.23 333.33
4366.76 231.86/
6987.19 609.10eh
4763.58 284.30/
5764.25 333.16
19
20
21
22
4125.36 225.49/
5177.84 334.94
5687.31 214.66b/
5115.18 201.45
4428.46 295.45/
5627.67 347.18
4425.16 223.31/
5026.34 198.74
4335.26 189.61/
5069.25 423.67
7061.15 175.07c/
7832.17 282.40eg
4513.06 197.33/
5624.51 300.39
4550.21 292.13/
5573.24 462.15
23
24
25
Each value represents means SD of four determinations.
aP < 0.05, bP < 0.01, cP < 0.001 compared to vehicle (0.25% DMSO).
dP < 0.05, eP < 0.01, fP < 0.001 compared to vehicle (0.25% DMSO) with 1.2 nM insulin.
i
gP < 0.05, hP < 0.01, P < 0.001 compared to the data without insulin.
P < 0.05 was taken as the level of significance.
Table 3
control
9.00
Effect of compound 10 on human platelet aggregation induced by AA and ADP
Glimepiride(20mg/kg)
8.50
Compound
IC50 (l
mol/L)a
Compound 10(2.5mg/kg)
8.00
Compound 10(7mg/kg)
AA
ADP
7.50
Compound 10(20mg/kg)
7.00
10
Asp
0.21
0.35
3.00
9.34
6.50
6.00
a
IC50 values were determined from direct regression curve analysis.
*
5.50
*
*
5.00
4.50
4.00
3.50
3.00
2.50
2.00
**
**
**
**
**
In summary, we have developed a novel class of sulfonylurea
and thiourea derivatives substituted with benzenesulfonamide
groups as potential hypoglycemic agents. These compounds in-
creased the insulin release of isolated rat pancreatic islets, im-
proved the insulin-stimulated glucose transport in adipocytes of
rats in vitro. Compound 10 with fused structure of glimepiride
and doltroban or S18886, showed remarkable activities to the
two in vitro models above. In vivo compound 10 has also shown
efficacy to lower plasma glucose level of fasted C57BL/6J mice. Fur-
thermore compound 10 had potent antiplatelet activity and
showed an excellent property to protect collagen–epinephrine-in-
**
**
**
**
**
**
**
**
**
**
**
0
0.5
1
2
4
6
time(h)
Figure 2. Effect of compound 10 on fasted plasma glucose in C57BL/6 J mice,
means SEM, n = 10, *P < 0.05,**P < 0.01, versus control.

1744
H. Zhang et al. / Bioorg. Med. Chem. Lett. 19 (2009) 1740–1744
17. Worth, N. F.; Berry, C. L.; Thomas, A. C.; Campbell, J. H. Atherosclerosis 2005,
Table 4
183, 65.
Effect of compound 10 in the collagen–epinephrine-induced cerebral thrombosis
18. Viles-Gonzalez, J. F.; Fuster, V.; Roberto, C.; Carolina, V.; Randolph, H.; Stefano,
C.; Anand, S. X.; Badimon, J. J. Eur. Heart J. 2005, 26, 1557.
19. Cayatte, A. J.; Du, Y.; Oliver-Krasinski, J.; Lavielle, G.; Verbeuren, T. J.; Cohen, R.
A. Arterioscler. Thromb. Vasc. Biol. 2000, 20, 1724.
20. Zuccollo, A.; Shi, C.; Mastroiann, R.; Maitland-Toolan, K. A.; Weisbrod, R. M.;
Zang, M.; Xu, S.; Jiang, B.; Oliver-Krasinski, J. M.; Cayatte, A. J.; Corda, S.;
Lavielle, G.; Verbeuren, T. J.; Cohen, R. A. Circulation 2005, 112, 3001.
21. Šebeková, K.; Eifert, Timo; Klassen, A.; Heidland, A.; Amann, K. Diabetes 2007,
56, 968.
mice model
Compound
Asp
10
Dose (mg/kg)
Survival rate (%)
60
10
20
40
63.6b
27.3a
45.5b
72.7b
Data represent the survival percent in a group of 11 mice.
aP < 0.05, bP < 0.01, compared with control.
22. Choi, S. W.; Benzie, F. F.; Ma, S. W.; Strain, J. J.; Hannigan, B. M. Free Radical Biol.
Med. 2008, 44, 1217.
23. Smith, E. F., 3rd; McDonald, J. Pharmacology 1988, 36, 340.
24. Englert, H. C.; Gerlanch, U.; Goegelein, H. J. Med. Chem. 2001, 44, 1085.
25. Selected spectroscopic data: Compound 10: IR (KBr): 3259, 2927, 1690, 1336,
Table 5
ꢀ
Main PK parameters of compound 10 in rats (x s, n = 6)
1161, 1090 cmÀ1 1H NMR (DMSO-d6, 300 MHz,d ppm): 7.37–7.88 (m, 8H, Ar-
;
Parameters
Value
H), 6.29 (d, 1H, NH-C6H10CH3), 3.32 (m, 1H,), 3.30 (q, 2H, CH2), 2.78 (t, 2H, CH2),
0.81–1.69(m, 12H,); MS (ESI, m/z): 558 ([M+H]+, base peak), 580 ([M+Na]+);
Anal. Calcd for C22H28BrN3O5S2: C 47.31, H 5.05, N 7.52; found: C 47.20, H 5.04,
N 7.65.
Dose iv/op (mg/kg)
t1/2 (iv, h)
t1/2 (op, h)
Vd (L/kg)
CL (L/h)
Tmax (h)
Cmax (mg/L)
AUCop (mg/L/h)
F (%)
2.5/5.0
1.91 0.65
1.77 0.21
0.78 0.36
0. 36 0. 22
0.83 0.13
3.33 0.80
10.04 2.43
52.69
26. The pancreatic tissue was removed from rats and digested with collagenase I
for 5–8 min at 38 °C for 4–5 times. Dispersed pancreatic cells were washed
with Hanks liquid (containing 10% fetal bovine serum). After centrifugation,
the isolated cells were cultured in RPMI 1640 medium with the addition of 10%
fetal bovine serum. Fibroblasts were taken out with iodoacetic acid incubated
for 48 h. Final incubation were carried out for 6 h at 37 °C with glucose
(25 mmol/L), test and standard compounds at different concentrations. The
insulin concentration in 1640 medium was measured by radioimmunoassay
method.
duced mice mortality. Although the mechanism by which com-
pound 10 acts is not well established, the preliminary pharmaco-
logical profile of compound 10 showed that it may be related to
TPr and might be useful in the treatment of diabetics with cardio-
vascular and nephropathy complications. Further evaluation of
compound 10 is underway.
27. Jerrold, M. O. Biochem. J. 1978, 172, 137.
28. Fat cells were separated from SD rats and identified by oil red O-staining.
0.5 ml adipocyte suspension (5 Â 105 cells/ml) was added into 2-ml
polypropylene plastic tube and incubated for 30 min at 37 °C in 5% CO2
incubator. Different concentrations of target compounds (0.25% DMSO) were
added in different experimental groups and incubated at 37 °C for 30 min. In
the presence or absence of 1.2 nM insulin the cells were incubated at 37 °C for
4 h. According to the different experimental groups, 2-Deoxy-
was added (final concentration 0.2 Ci/ml) and after incubated for 1.5 h, ice-
D
-[1-3H]glucose
l
cold 4 °C dimethyl silicone was added immediately. After centrifugation at
10,000 rpm for 2 min, lower layer culture was removed. One hundred
microliters 5% SDS was added to crack the upper fat clumps. After 0.5 h, the
lysis was added to scintillation vial with 5 ml scintillation liquid (4.0 g PPO,
0.4 g POPOP dissolved in 1 L xylene), standed in dark for 12 h and the radiation
intensity (Bq) of 3H was measured by liquid scintillation counting.
Acknowledgment
The work was supported by the Hi-tech Research and Develop-
ment Program of China (No. 2003AA2Z3530).
29. Fasted C57BL/6 J mice were dosed daily by oral gavage for 3 days. Treatments
were compound 10 (20, 7, and 2.5 mg/kg), glimepiride (20 mg/kg) or vehicle
(0.5% carboxymethylcellulose). On day 5, mice were weighed and the blood
samples were collected from the cut tip of the tail vein at 0, 0.5, 1, 2, 4, and 6 h
after dosing. Samples were assayed for glucose. Plasma glucose was measured
using the glucose oxidase Trinder method.
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into plastic vessels containing 0.1 volume of 3.8% aqueous sodium citrate.
Platelet-rich plasma (PRP) was then prepared by centrifugation at 1000 rpm for
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ll by using PPP. By using ADP
(5 lmol/L) and AA (200 lmol/L) as an aggregating inducer, respectively,
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l
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lg
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8 lg
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