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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30. Blood was collected from the cubitus artery of healthy volunteers, who were
limosis and avoiding treatment with antithrombotic drugs 2 weeks before test,
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
10 min. Platelet-poor plasma (PPP) was prepared from the remaining blood
after removing PRP, by centrifugation at 3000 rpm for 10 min. PRP was
adjusted to a concentration of 6.0 Â 10À5 cells/
ll by using PPP. By using ADP
(5 lmol/L) and AA (200 lmol/L) as an aggregating inducer, respectively,
platelet aggregation of Aspirin (ASP) solution or the test compound solution
(final concentration of ASP and compound 10: 100, 10, 1, and 0.1 mol/L) was
l
assayed on a four-channel aggregometer (LBY-NJ2 type, Plysion Co., Ltd).
31. Chinese Kunming mice weighing about 30 g were grouped after overnight
fasting (n = 11). Mice were dosed once daily by oral gavage for 7 days.
Treatments were compound 10 (40, 20, and 10 mg/kg), ASP (60 mg/kg) or
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lg
collagen and epinephrine were injected intravenously for 20 s. The
8 lg
protective effects were calculated by measuring the incidence of death within
15 min of injection. Results are expressed as percent survival.
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