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E. Kim et al. / Bioorg. Med. Chem. Lett. 18 (2008) 4993–4996
Table 4
In vivo data of compound (S)-5b in db/db micea
Compound
Reduction of plasma glucoseb (%)
D
BWc (g)
0.3
ED30
0.03d
0.1
0.3
1.0
3.0
0.03
0.1
1.0
3.0
(mg/kg)
Rosiglitazone
Muraglitazar
(S)-5b
—
—
14
17
4
37
37
11
63
—
56
54
77
68
78
—
—
—
+11.2
+10.5
+10.5
+12.4
+2.4
+11.1
+12.7
+12.6
+12.5
+14.5
+13.7
+13.9
+16.1
—
0.210
0.411
0.067
—
Lean
db/db
—
+9.0
—
a
Male db/db mice (7-week-old) and lean mice were dosed daily for 28 days by oral with the indicated doses of test compound.
Reduction of plasma glucose was calculated as the percentage reduction with respect to the non-fasting glucose value of control after 28 days of oral dosing.
Body weight change in db/db mice after 28 days of oral dosing.
Dose in mg/kg unit.
b
c
d
PPAR
headpiece,18 compounds 13c and 13d showed remarkably in-
creased PPAR activity and significantly reduced PPAR activity
-ethoxy-b-phenyl propanoic acid headpiece-
a activity. With the 2-dimethyl-phenoxy propanoic acid
References and notes
1. Moller, D. E. Nature 2001, 414, 821.
a
c
2. Han, H. O.; Kim, S. H.; Kim, K.-H.; Hur, G.-C.; Yim, H. J.; Chung, H.-K.; Woo, S. H.;
Koo, K. D.; Lee, C.-S.; Koh, J. S.; Kim, G. T. Bioorg. Med. Chem. Lett. 2007, 17, 937.
3. Willson, T. M.; Brown, P. J.; Sternbach, D. D.; Henke, B. R. J. Med. Chem. 2000, 43,
527.
compared with
a
based 5b.
After determining the in vitro properties of the aryl-tetrahydro-
pyridine compounds, compound (S)-5b was selected for the phar-
macokinetic (PK) study in rats, and the in vivo experiment in male
db/db mice for a further evaluation.19 The PK parameters of (S)-5b
in Sprague–Dawley (SD) rats were satisfactory, as shown in Table 3.
In the PK study, compound (S)-5b exhibited an apparently long-
er half-life than rosiglitazone and muraglitazar (Fig. 1), and had
good water solubility (3.26 mg/mL, pH = 6.8), which may contrib-
ute to the excellent oral bioavailability (ꢂ100%). Because the
aryl-tetrahydropyridine compound (S)-5b has both acid (–COOH)
and amino (–NRR0) functional groups in the molecule, it has possi-
ble zwitterionic character under neutral conditions (pH = 7.0). As a
result, (S)-5b showed good water solubility and stability over all
pH region, which probably is the reason for the satisfactory PK
profile.
Compound (S)-5b was evaluated in the db/db mouse model,
using rosiglitazone and muraglitazar (Fig. 1) for comparison.20
When the db/db mice were treated orally by 0.03, 0.1, 0.3, and
1.0 mg/kg for 28 days, (S)-5b showed dose-dependent decrease
of blood glucose level (Table 4). Treatment with 1.0 mg/kg of (S)-
5b showed a glucose reduction of 77% compared with that of
68% with 3.0 mg/kg of rosiglitazone. A glucose lowering effect
was also observed at a dose of 0.1 mg/kg of (S)-5b. The ED30 of
(S)-5b was determined to be 0.067 mg/kg, which is significantly
lower than the ED30 of rosiglitazone (0.210 mg/kg) and muraglita-
zar (0.411 mg/kg). Consequently, compound (S)-5b displayed a
better glucose lowering effect than rosiglitazone and muraglitazar
based on the oral dose, whereas it also caused a similar increase in
body weight to muraglitazar.
4. Fruchart, J.-C.; Duriez, P.; Staels, B. Curr. Opin. Lipidol. 1999, 10, 245.
5. Bloomgarden, Z. T. Diabetes Care 2005, 28, 2.
6. Murakami, K.; Tobe, K.; Ide, T.; Mochizuki, T.; Ohashi, M.; Akanuma, Y.; Yazaki,
Y.; Kadowaki, T. Diabetes 1998, 47, 1841.
7. Khare, A. B.; McKenna, C. E. Synthesis 1991, 5, 405.
8. Haigh, D.; Birrell, H. C.; Cantello, B. C. C.; Hindley, R. M.; Ramaswamy, A.; Rami,
H. K.; Stevens, N. C. Tetrahedron: Asymmetry 1999, 10, 1335.
9. Gotteland, J.-P.; Loubat, C.; Planty, B.; Junquéro, D.; Delhon, A.; Halazy, S. Bioorg.
Med. Chem. Lett. 1998, 8, 1337.
10. Wustrow, D. J.; Wise, L. D. Synthesis 1991, 11, 993.
11. Eastwood, P. R. Tetrahedron Lett. 2000, 41, 3705.
12. In vitro transient transactivation assays: the ligand binding domains (LBD) of
the human PPARa/c receptors were fused to the DNA binding domain (DBD) of
the yeast transcription factor GAL4. The CV-1 cells were transiently transfected
with an expression vector for the respective PPAR chimera along with a
reporter construct containing five copies of the GAL4DNA binding site and pRL-
TK as a control vector (Promega, WI). The test compounds were dissolved in
DMSO and diluted 1:1000 in DMEM supplemented with 10% fetal bovine
serum (FBS) and 0.1 mM MEM non-essential amino acid (MEAA). The cells
were treated with the test compounds in triplicate for 24 h and subjected to a
dual luciferase assay using the Dual-Glo luciferase reagent (Promega, WI). The
increases in luminescence were measured using a microplate reader (Lmax
II384, Molecular Devices, CA). EC50 values were calculated using the Hill 4-
parametric equation in SigmaPlo t 4.0 (SPSS, IL).
13. Kasuga, J.-I.; Hashimoto, Y.; Miyachi, H. Bioorg. Med. Chem. Lett. 2006, 16, 771.
14. Haigh, D.; Allen, G.; Birrell, H. C.; Buckle, D. R.; Cantello, B. C. C.; Eggleston, D.
S.; Haltiwanger, R. C.; Holder, J. C.; Lister, C. A.; Pinto, I. L.; Rami, H. K.; Sime, J.
T.; Smith, S. A.; Sweeney, J. D. Bioorg. Med. Chem. 1999, 7, 821.
15. Haigh, D.; Birrell, H. C.; Cantello, B. C. C.; Eggleston, D. S.; Haltiwanger, R. C.;
Hindley, R. M.; Ramaswamy, A.; Stevens, N. C. Tetrahedron: Asymmetry 1999,
10, 1353.
16. The difference values of the PPARa/c agonistic activity between the racemate
5r and the (S)-isomer, (S)-5r were not enough significative to conclude as the
latter was a better one. Even though we do not describe in this letter, another
(S)-isomers of compounds 5 showed a tendency to have a better activity than
the comparative racemates.
17. Brooks, D. A. W.O. 02/16331 A1, 2002.
18. Desai, R. C.; Metzger, E.; Santini, C.; Meinke, P. T.; Heck, J. V.; Berger, J. P.;
Macnaul, K. L.; Cai, T.-Q.; Wright, S. D.; Agrawal, A.; Moller, D. E.; Sahoo, S. P.
Bioorg. Med. Chem. Lett. 2006, 16, 1673.
In summary, this letter reported the synthesis and in vitro char-
acterization of a novel series of potent and selective PPARa/c dual
agonists. Compound (S)-5b showed an excellent in vivo profile in
the db/db mouse model of type II diabetes, and also shows desir-
able pharmacokinetic properties. A further evaluation of com-
pound (S)-5b is currently underway.
19. In vivo study: It was evaluated using two reference substances, rosiglitazone,
which is marketed, and muraglitazar, which has a different acid headpiece
from compound (S)-5b. Tesaglitazar was used not in in vivo study but in in
vitro assay as a reference substance.
20. In vivo db/db mice study: male homozygous db/db mice were obtained from
Japan SLC, Inc. (Shizuoka Prefecture, Japan). In order to evaluate the effect of
the test compound, 7-week-old animals were acclimated for 1 week, and
allocated to the corresponding groups (n = 7/group) according to the plasma
glucose level and weight. The mice were treated once daily by an oral gavage
with the test compound, muraglitazar, rosiglitazone, or an equivalent amount
of 0.5% MC for 28 days. The decreases in the plasma glucose levels were
calculated as the percentage change from those of the controls.
Acknowledgment
This study was supported by a grant of the Korea Health 21 R&D
Project, Republic of Korea (A 020600).