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CPT1A
Vehicle 2 uM 0.2 uM 2 uM 20 uM
6,90
7
6
5
4
3
2
1
0
3,59
2,36
1,68
0,96
DMSO
GW7647 GW7647 + 4g GW7647+ 4g GW7647+ 4g
Figure 5. Gene repression profile of 4g. HepG2 cells were treated with 2
lM of GW7647, without or with the addition of 4g from 0.2 to 20
lM. RTqPCR was performed to
measure CPT1A mRNA levels.
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the impact of selected compounds 4c, 4e, and 4g on PPARa expres-
sion, and measurement of mRNA concentration was done as a
quantitative analysis real-time reverse transcription polymerase
chain reaction (RTqPCR). When applied alone (150 lM), the three
compounds were not able to produce gene activation; however,
when the cells were treated with a combination of GW7647
(2 lM) and 4c, 4e, and 4g, the levels of mRNA quantified were con-
siderably reduced, so showing an interesting inhibition of CPT1A
activation (Fig. 4). This trend was studied also at lower concentra-
tions of compound 4g, which was able to decrease gene induction
from 0.2 to 20 lM in a dose-dependent manner, even if the inhib-
itory effect results quite lower with regarding higher concentra-
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In conclusion, the purpose of the present study was to deter-
mine the effect of some N-(phenylsulfonyl)amides on PPARa activ-
ity, by using a transient transfection system in human embryonic
kidney 293 cells. We have identified three compounds that inhibit
in a dose-dependent manner the activation of the receptor in the
19. Data for selected N-(phenylsulfonyl)amides. 2-[(5-Chloro-1,3-benzothiazol-2-
yl)thio]-N-(phenylsulfonyl)pentanamide (4c): white needles, 24% yield, mp 123–
124 °C. IR (KBr) 1699, 1354 and 1186 cmÀ1 1H NMR (CDCl3) d 0.91 (t, 3H,
;
presence of a full agonist. The repressive effect on PPARa-respon-
J = 7.5 Hz, CH3CH2), 1.38–1.65 (m, 2H, CH2CH3), 1.85–2.22 (m, 2H, CH2CH), 4.16
(t, 1H, J = 7.5 Hz, CH), 7.39 (dd, 1H, J = 8.7 Hz, J = 2.1 Hz, CH arom), 7.45–7.62
(m, 3H, CH arom), 7.70 (d, 1H, J = 8.7 Hz, CH arom), 7.90 (d, 1H, J = 2.1 Hz, CH
arom), 7.92–7.97 (m, 2H, CH arom), 11.68 (b s, 1H, NH); 13C NMR (CDCl3) d 13.7
(CH3CH2), 20.5 (CH2CH3), 31.2 (CH2CH), 50.2 (CH), 121.6, 122.1, 126.2, 128.3
and 129.0 (CH arom), 133.2 and 133.4 (C arom), 134.0 (CH arom), 138.6, 152.5
and 168.4 (C arom), 169.1 (C@O). Anal. CHN: Calcd for C18H17ClN2O3S3: C,
49.02; H, 3.89; N, 6.35. Found: C, 49.04; H, 3.87; N, 6.32. 2-[(5-Chloro-1,3-
benzothiazol-2-yl)thio]-N-(phenylsulfonyl) hexanamide (4e): white solid, 71%
sive gene CPT1A was examined in detail for compound 4g.
Acknowledgment
Support for this study was provided by University ‘G. d’Annun-
zio’ of Chieti local grants. The authors gratefully acknowledge Dr.
Antonio Moschetta for his precious scientific help and Dr. David
J. Mangeldorf (Dallas, TX) for the PPAR plasmids.
yield, mp 128–129 °C. IR (KBr): 3432, 1706, 1451, 1361, 1187 cmÀ1 1H NMR
;
(CDCl3) d 0.86 (t, 3H, J = 7.2 Hz, CH3CH2), 1.24–1.49 (m, 4H, CH2), 1.63–2.17 (m,
2H, CH2), 4.14 (t, 1H, J = 7.2 Hz, CHS), 7.37 (dd, 1H, J = 8.7 Hz, J = 2.1 Hz, CH Ar),
7.41–7.62 (m, 3H, CH Ar), 7.71 (d, 1H, J = 8.7 Hz, CH Ar), 7.89–7.92 (m, 2H, CH
Ar), 7.95 (d, 1H, J = 2.1 Hz, CH Ar), 11.67 (b s, 1H, NH); 13C NMR (CDCl3) d 13.9
(CH3CH2), 22.4, 29.0 and 29.3 (CH2), 50.4 (CH), 121.7, 122.1, 126.2, 128.3 and
129.0 (CH Ar), 133.2 and 133.4 (C Ar), 134.0 (CH Ar), 138.6, 152.4 and 168.4 (C
Ar), 169.1 (C@O). Anal. CHN: Calcd for C19H19ClN2O3S3: C, 50.15; H, 4.21; N,
6.16. Found: C, 50.17; H, 4.22; N, 6.13. 2-[(5-Chloro-1,3-benzothiazol-2-yl)thio]-
2-phenyl-N-(phenylsulfonyl)acetamide (4g): white solid, 30% yield, mp 140–
Supplementary data
Supplementary data associated with this article can be found, in
142 °C. IR (KBr) 3256, 1709, 1449, 1422, 1363, 1183 cmÀ1 1H NMR (CDCl3) d
;
References and notes
5.43 (s, 1H, CHS), 7.33–7.60 (m, 8H, CH arom), 7.67 (d, 1H, J = 8.4 Hz, CH arom),
7.78 (d, 1H, J = 1.8 Hz, CH arom), 7.91–7.97 (m, 3H, CH arom), 11.05 (b s, 1H,
NH); 13C NMR (CDCl3) d 54.6 (CHS), 121.8, 122.1, 126.0, 126.6, 128.5, 129.0,
129.1 and 129.3 (CH arom), 132.2, 133.0 and 133.5 (C arom), 134.1 (CH arom),
138.3, 152.6 and 166.7 (C arom), 167.9 (C@O). Anal. CHN: Calcd for
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