Design and synthesis of novel indole-chalcone fibrates as lipid lowering agents

Article abstract of DOI:10.1016/j.ejmech.2014.04.085

A series of novel indole-chalcone fibrates were synthesized and their hypolipidemic activity was evaluated in triton WR-1339 induced hyperlipidemic rat model. Preliminary studies indicated that the hybrids 19, 24 and 29 exhibited potent in vitro antioxidant and significant in vivo antidyslipidemic effects. Our results suggest that these new hybrid architectures may serve as promising leads for the development of next generation lipid lowering agents.

Full text of DOI:10.1016/j.ejmech.2014.04.085

Accepted Manuscript
Design and synthesis of novel indole-chalcone fibrates as lipid lowering agents
Koneni V. Sashidhara , Ranga Prasad Dodda , Ravi Sonkar , Gopala Reddy Palnati ,
Gitika Bhatia
PII:
S0223-5234(14)00456-5
10.1016/j.ejmech.2014.04.085
EJMECH 6998
DOI:
Reference:
To appear in: European Journal of Medicinal Chemistry
Received Date: 22 January 2014
Revised Date: 2 April 2014
Accepted Date: 2 April 2014
Please cite this article as: K.V. Sashidhara, R.P. Dodda, R. Sonkar, G.R. Palnati, G. Bhatia, Design
and synthesis of novel indole-chalcone fibrates as lipid lowering agents, European Journal of Medicinal
Chemistry (2014), doi: 10.1016/j.ejmech.2014.04.085.
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ACCEPTED MANUSCRIPT
Graphical abstract
Design and synthesis of novel indole-chalcone fibrates as lipid
lowering agents
Koneni V. Sashidhara *, Ranga Prasad Dodda, Ravi Sonkar, Gopala Reddy Palnati, Gitika
Bhatia
The design and synthesis of new hybrid architecture in the search of novel lipid lowering
agents are described.

ACCEPTED MANUSCRIPT
Design and synthesis of novel indole-chalcone fibrates as lipid
lowering agents*
a,
a
b
a
Koneni V. Sashidhara *, Ranga Prasad Dodda , Ravi Sonkar , Gopala Reddy Palnati ,
Gitika Bhatia
b
a
Medicinal and Process Chemistry Division , CSIR-Central Drug Research Institute, BS-
1
0/1, Sector 10, Jankipuram extension, Sitapur Road, Lucknow 226031, India.
b
Biochemistry Division , CSIR-Central Drug Research Institute, BS-10/1, Sector 10,
Jankipuram extension, Sitapur Road, Lucknow 226031, India.
Abstract
A series of novel indole-chalcone fibrates were synthesized and their hypolipidemic activity
was evaluated in triton WR-1339 induced hyperlipidemic rat model. Preliminary studies
indicated that the hybrids 19, 24 and 29 exhibited potent in vitro antioxidant and significant
in vivo antidyslipidemic effects. Our results suggest that these new hybrid architectures may
serve as promising leads for the development of next generation lipid lowering agents.
Keywords: Indole-chalcone fibrates, antidyslipidemic activity, triton model, LPL activity,
antioxidant activity.
*
*
Part XXVIII in the series, “Advances in drug design and discovery”.
Corresponding author. Tel.: +91 9919317940; Fax: +91-0522-2771942.
Email: sashidhar123@gmail.com, kv_sashidhara@cdri.res.in.
1

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1
. Introduction
Coronary heart disease (CHD) is the dominant cause of morbidity and mortality in the
industrialized nations [1-2]. Among the predominant risk factors for CHD are high levels of
low-density lipoprotein cholesterol (LDL-C), triglycerides and low levels of high-density
lipoprotein cholesterol (HDL-C) [3]. An improvement in the levels of these risk factors
would reduce the risk for developing CHD. Currently, the most common method to treat
dyslipidemia is the use of statins, which are HMG-CoA reductase inhibitors. The widespread
clinical use of the statins is accompanied by potential dose-limiting hepatotoxicity and
myotoxicity, which may be the consequence of reduced levels of essential isoprenoid
precursors, the antioxidant ubiquinone, or dolichols [4]. Cerivastatin, one of the second
generation statins, was withdrawn from the world market in 2001, due to its adverse effects
[
5]. The fibrate class of lipid-lowering drugs such as fenofibrate and gemfibrozil (Figure. 1)
are selective activators of alpha-isotype of the receptors peroxisome proliferator activated
receptor (PPAR) [6, 7]. These drugs lower triglyceride levels and increase HDL cholesterol
levels in hyperlipidemic patients [8] and reduce the risk of coronary heart disease in patients
with low HDL cholesterol levels [9]. In recent years the design of dual-target hypoglycemic
agents that activate glucokinase and PPARγ have also been reported for treating Type 2
diabetes mellitus [10]. Recent studies outline the role of oxidative stress in the pathogenesis
and development of various diseases including atherosclerosis, diabetes mellitus,
hypertension, and coronary heart disease [11]. Oxidative stress is characterized by an
imbalance between reactive oxygen species (ROS) and antioxidant systems [12]. So a single-
targeted therapeutic approach in multifactorial disorders such as those mentioned above is
mostly considered inadequate [13, 14]. Thus molecules endowed with both antidyslipidemic
and antioxidant properties are in great demand.
Molecular hybridization is a strategy of rational design of new ligands or prototypes based on
the recognition of pharmacophoric subunits in the molecular structure of two or more known
bioactive derivatives [15]. Owning to its wide range of pharmacological activities, indole and
their derivatives has been a subject of intense investigations [16-20]. Fluvastatin (Figure 1),
which includes an indole moiety, is a synthetic member of the statin class of drug, used to
lower cholesterol and prevent cardiovascular disease. Our group recently reported a series of
coumarin-bisindole hybrids, which have shown significant antihyperlipidemic activity [21]
2

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(Figure 1). On the other hand, chalcones are important compounds that are present in many
naturally occurring products. Their pharmaceutical potential is due to their radical-
scavenging [22, 23], antitumor [24, 25], anti-inflammatory [26] and neuro-protective
properties [27]. Furthermore, recent literature survey revealed that synthetic chalcone
'
derivatives, DE027 and 4 , 4-dichlorochalcone (Figure 1), exhibited potential hypolipidemic
activities [28]. In our previous reports, we disclosed the indole based fibrates and coumarin-
chalcone fibrates as potent antidyslipidemic agents [29, 30] (Figure 2).
These above interesting findings and our continuous quest to identify more potent analogues,
led to the molecular hybridization of indole, chalcone and fibrate to integrate them in one
molecular platform to generate a new hybrid architecture with the aim of exploring the
impact of such modification on the hypolipidemic profile [31, 32] (Figure 2). In this paper we
report the synthesis and evaluation of novel indole-chalcone fibrates as new class of
antidyslipidemic agents. Figure 1 shows some representative chemical structures of
compounds possessing indole, fibrate and chalcone framework with lipid lowering activities.
2
. Chemistry
The synthesis of the designed compounds are outlined in Scheme 1. The commercially
available indole and substituted indoles (1-4) underwent Vilsmeier Haack reaction [33] in
presence of POCl and DMF to afford corresponding indole-3-carbaldehydes (5-8). On the
3
other hand, the reaction of 4-hydroxyacetophenone with the appropriate bromo esters in the
presence of K CO3 in acetonitrile under reflux conditions furnished substituted
2
acetophenones (9-12) in quantitative yield. The subsequent Claisen-Schmidt condensation of
these acetophenones (9-12) with indole-3- carbaldehydes (5-8) in the presence of catalytic
amount of piperidine in MeOH or EtOH under reflux conditions [34] yielded the desired
indole-chalcone fibrates 13-21 and 22-30, respectively and the trans double bond was
obtained exclusively (J= 15-16Hz). When using methanol as the solvent and piperdine as a
base a significant amount of transesterification product leading to compounds 13-21 were
1
13
obtained [35].The structures of the compounds were substantiated by H NMR, C NMR,
mass spectrometry and IR spectroscopy.
3
. Pharmacology
3
.1. Animals used.
3

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Rats (Charles Foster strain, male, adult, body weight 200–225g) were kept in a room with
o
controlled temperature (25–26 C), humidity (60–80%) and12/12 h light/dark cycle (light on
from 8.00 A.M. to 8.00 P.M.) under hygienic conditions. Animals, which were acclimatized
for one week before starting the experiment, had free access to the normal diet and water.
3
.2. Lipid lowering and post heparin lipolytic activity
Rats were divided into twelve groups control, triton induced, triton plus 13-30 and
gemfibrozil (100 mg/kg) treated groups, containing six rats in each group. In this experiment
of 18h, hyperlipidemia was developed by administration of triton WR-1339 (Sigma chemical
company, St. Louis, MO, USA) at a dose of 400 mg/kg body weight intraperitoneally to
animals of all the groups except the control. These derivatives were macerated with gum
acacia (0.2% w/v), suspended in water and fed simultaneously with triton at a dose of 100
mg/kg p.o. to the animals of treated group and the diet being withdrawn. Animals of control
and triton group without treatment with hybrid compounds were given same amount of gum
acacia suspension (vehicle). After 18h of treatment the animals were anaesthetized with
thiopentone solution (50 mg/kg b.w.) prepared in normal saline and then 1.0 ml blood was
withdrawn from retro-orbital sinus using glass capillary in EDTA coated eppendorf tube (3.0
o
mg/ml blood). The blood was centrifuged (at 2500g) at 4 C for 10 min and plasma was
separated. Plasma was diluted with normal saline (ratio of 1:3) and used for analysis of total
cholesterol (TC), triglycerides (TG) and phospholipids (PL) by standard enzymatic methods
[
36] and post heparin lipolytic activity (PHLA) were assayed [37] using spectrophotometer,
Beckmann auto-analyzer and standard kits purchased from Beckmann Coulter International,
USA.
3
.3. Lipoprotein measurement in blood plasma of triton induced hyperlipidemic rats.
Plasma was fractionated into very low density lipoprotein (VLDL), low density lipoprotein
LDL) and high density lipoprotein (HDL) by poly anionic precipitation methods. Plasma and
(
lipoproteins were analyzed for their total cholesterol (TC), phospholipid (PL), and
triglyceride (TG) by standard procedures reported earlier [38].
3
.4. Lipoprotein lipase activity in liver of triton induced hyperlipidemic rats.
Liver was homogenized (10%, w/v) in cold 100 mM phosphate buffer pH 7.2 and used for
the assay of total lipolytic activity of lipoprotein lipase (LPL) [39].
3
.5. Antioxidant activity (generation of free radicals).
4

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Superoxide anions (O2-) were generated enzymatically [40] by xanthine (160 mM), xanthine
oxidase (0.04 U) and nitroblue tetrazolium (320 µM) in absence or presence of compounds
(200µg/ml) in 100mM phosphate buffer (pH 8.2). Fractions were sonicated well in phosphate
o
buffer before use. The reaction mixtures were incubated at 37 C and after 30 min the
reaction was stopped by adding 0.5 mL glacial acetic acid. The amount of formazone formed
was measured at 560 nm on a spectrophotometer. Percentage inhibition was calculated taking
3
-1
-1
absorption coefficient of formazone as 7.2 x10 M 1cm . In another set of experiment, an
•
effect of compounds on generation of hydroxyl radicals ( OH) was also studied by non-
•
enzymic reactants [41]. Briefly ( OH) were generated in a non-enzymic system comprised of
deoxy ribose (2.8 mM), FeSO .7H O (2 mM), sodium ascorbate (2.0 mM) and H O (2.8
4
2
2
2
mM) in 50 µM KH PO buffer, pH 7.4 to a final volume of 2.5 mL. The above reaction
2
4
o
mixtures in the absence or presence of compounds (200 µg/mL) were incubated at 37 C for
0 min. Reference samples and reagent blanks were also run simultaneously.
9
Malondialdehyde (MDA) content in both experimental and reference samples were estimated
spectrophotometrically by thiobarbituric acid method as mentioned above [42].
3
.6. Statistical evaluation
Data were analyzed using student’s t-test. The hyperlipidemic groups were compared with
control drug treated groups. P<0.05 was considered to be significant.
4
. Results and Discussion
4
.1. The lipid lowering activity of indole-chalcone fibrates
The antidyslipidemic activity of novel indole-chalcone fibrates was evaluated in vivo in
Triton model. Hyperlipidemia was established after Triton WR-1339 administration,
(liberation of cholesterol from the liver induced by Triton WR-1339 triggers the hepatic
synthesis of cholesterol) as evidenced by an increase in the plasma level of total cholesterol
TC (4.0 fold), phospholipids PL (3.3 fold) and triglyceride TG (2.6 fold) and plasma post
heparin lipolytic activity (PHLA) (30%) as compared to control). Hyperlipidemic rats were
treated with indole chalcone fibrates at a dose of 100 mg/kg p.o. reversed the plasma levels of
lipids with varying extents. Compounds 19, 24 and 29 exhibited excellent antidyslipidemic
effect. Specifically, compounds 24 and 29 significantly lowered the TC, PL, and TG by 31%,
3
2%, and 30%, and 32%, 33%, and 30% respectively, where as compound 19 lowered them
by 26%, 24%, and 27% respectively. Compounds 15, 23 and 28 showed good hypolipidemic
5

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activity. Under the same experimental conditions and at the same dose 100 mg/kg
gemfibrozil (standard hypolipidemic drug) decreased levels of TC, PL and TG in plasma by
3
1%, 33% and 33%, respectively. Compounds 19, 24 and 29 showed significant reversal of
PHLA in plasma of hyperlipidemic rats by 16%, 19% and 20% respectively, comparable to
gemfibrozil, which caused 19% reversal of activity of this enzyme as compared to control
group and the results are summarised in Table 1.
These results strongly demonstrate that novel indole-chalcone fibrates displayed enhanced
antidyslipidemic activity than our previously developed multifunctional hybrids. The rational
designing and preliminary structure activity relationship (SAR) studies of new synthesized
compounds are depicted in Figure 2.
4
.2. Effect of compounds 19, 24 and 29 on lipid lowering at different doses in triton induced
hyperlipidemic rat.
After the confirmation of most active compounds in primary screening the efficacy of active
compounds 19, 24 and 29 at different doses in 50 to 150 mg/kg body weight was studied.
Compound 19 lowered the TC by 14% to 29%, PL by 17% to 24% and TG by 15% to 29%.
Compound 24 lowered TC by 22% to 31%, PL by 20% to 33% and TG by 20% to 30%.
Similarly compound 29 lowered the TC by 24% to 35%, PL by 26% to 34% and TG by 25%
to 33% respectively. The results are summarised in Figure 3.
4
.3. Effect of compounds 19, 24 and 29 on plasma lipoprotein lipids in triton induced
hyperlipidemic rats.
Having identified compounds 19, 24 and 29 as most potent in the primary screening, we
further tested for their effect on plasma lipoprotein lipids. As shown in Figure 4, the analysis
of plasma of triton administered hyperlipidemic rats showed marked increase in the level of
lipoprotein lipids and these effects were pronounced for VLDL and LDL followed by a
decrease in HDL as compared to control rats. Interestingly, the treatment with compounds 19,
2
4 and 29 significantly reversed the levels of VLDL, LDL and HDL, the complete lipid
profile results are shown in Figure 4.
4
.4. Effect of compounds 19, 24 and 29 on liver-LPL activity in triton induced hyperlipidemic
rats.
Additionally the compounds 19, 24 and 29 were studied for liver-LPL activity in triton
induced hyperlipidemic rats. Administration of triton in rats markedly decreases the LPL
6

ACCEPTED MANUSCRIPT
activity in liver as shown in Figure 5. However, the administration of compounds 19, 24 and
2
9 significantly increased the LPL activity. (Figure 5). The results were comparable with the
standard drug gemfibrozil, which at the same dose (100mg/kg) caused marked increase in the
LPL activity.
4
.5. Effect of compounds 13-30 on superoxide anions, hydroxyl radicals and lipid-
peroxidation
Further, the antioxidant activity of indole-chalcone fibrates 13-30 at 200µg/mL were
•
evaluated by generating free radicals (superoxide ions (O2-), hydroxyl radicals ( OH),
microsomal lipid peroxidation) in vitro in the absence and presence of these compounds. As
shown in Figure 6, compounds 15, 19, 23, 24 and 29 significantly suppressed the superoxide
ions by 26%, 27%, 33%, 31% and 36%, hydroxyl radicals by 32%, 29%, 38%, 37% and 41%
and microsomal lipid peroxidation by 30%, 28%, 36%, 35% and 40% respectively.
Therefore, it is evident that these compounds have strong antioxidant properties. The standard
drug alloperinol, showed 49% inhibition in superoxide ions, whereas mannitol and α-
tocopherol, showed 53% and 46% inhibition of hydroxyl ions and microsomal lipid
peroxidation, respectively at same experimental conditions. The scavenging potential of the
remaining compounds was moderate.
5
. Conclusion
In summary, a novel class of indole-chalcone fibrates were synthesized and evaluated for
their hypolipidemic activity. Compounds 19, 24 and 29 decreased the concentrations of TC,
PL and TG of hyperlipidemic rats. In addition, these compounds significantly reversed the
levels of VLDL, LDL and HDL and also increased the LPL activity. Furthermore, the active
hybrids displayed excellent in vitro antioxidant properties. From these results, it can be
concluded that this class of hybrids have great potential as a new class of antidyslipidemic
agents. Further studies to demonstrate its molecular mechanism and optimization of these
hybrids are required and our research efforts in this direction are in progress.
6
. Experimental
6
.1. General Information:
7

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All reagents were commercial and were used without further purification. Chromatography
was carried on silica gel (60-120 and 100-200 mesh). All reactions were monitored by TLC;
1
silica gel plates with fluorescence F254 were used. Melting points were uncorrected. The H
1
3
NMR and C NMR spectra were determined on 200, 300, 400 MHz and 50, 75, 100 MHz,
respectively, using CDCl , CD OD and DMSO-d as solvents and TMS as internal standard.
3
3
6
All chemical shifts were given in ppm and multiplicity (s = singlet, brs = broad singlet, d =
doublet, brd = broad doublet, dd = double doublet, t = triplet, q = quartet, m = multiplet). IR
-1
spectra were recorded on in the range of 500~4000 cm .
6
.2. General synthetic procedure for preparation of different indole-3-carbaldehydes (5-8):
Different indole-3-carbaldehyde substrates were prepared according to the reported method
32].
[
6
.3. General synthetic procedure for preparation of substituted acetophenone compounds (9-
3):
1
An oven dried 50 mL round bottom flask was charged with 4-hydroxy acetophenone (1.0
equiv.), substituted bromoester compound (1.1 equiv.) and acetonitrile (25 mL). K CO (10.0
2
3
0
equiv.) was then added, and the reaction mixture was heated to 90 C for 4 h. The acetonitrile
was evaporated in vacuum, and the residue was purified by silica gel (100-200) column
chromatography (20% EtoAc/Hexane) to afford ester compounds 5-8 in good yields.
6
.3.1. Ethyl 2-(4-acetylphenoxy) propionate (9):
–
1
White crystalline solid; yield: 92%; mp: 147-149 ºC; IR(KBr,cm ): 2925, 2352, 1677, 1566,
1
1
2
3
207, 1073, 760; H NMR (CDCl , 300 MHz) δ :7.93 (d, J = 8.7 Hz, 2H ), 6.91(d, J = 8.8 Hz,
3
H), 4.83 (q , J = 6.6 Hz, 1H), 4.23 (q , J = 7.1 Hz, 2H), 2.55 (s, 3H), 1.66 (d, J = 6.7 Hz,
+
H), 1.26 (t , J = 7.1 Hz, 3H); ESI-MS (m/z): 237 (M+H) .
6
.3.2. Ethyl 2-(4-acetylphenoxy) butanoate (10):
–
1
White crystalline solid; yield: 93%; mp: 139-141 ºC; IR(KBr,cm ): 2954, 2356, 1679, 1568,
1
1
2
207, 1069, 770; H NMR (CDCl , 400 MHz) δ :7.92 (d, J = 8.8 Hz, 2H ) 6.92(d, J = 8.8 Hz,
3
H) 4.63 (t , J = 6.1 Hz, 1H) 4.22 (q , J = 7.1 Hz, 2H) 2.55 (s, 3H) 2.05-1.98 (m, 2H) 1.24 (t,
+
J = 7.1 Hz, 3H); ESI-MS (m/z): 251 (M+H) .
6
.3.3. Ethyl 2-(4-acetylphenoxy)-2-methylpropanoate (11):
8

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–
1
1
Colorless oil; yield: 95%; IR(KBr,cm ): 2935, 2359, 1665, 1576, 1207, 1073, 778; H NMR
(
CDCl , 400 MHz) δ :7.87 (d, J = 8.9 Hz, 2H ), 6.83(d, J = 8.9 Hz, 2H), 4.22 (q , J = 7.1 Hz,
3
+
2
H), 2.54 (s, 3H), 1.65 (s, 3H), 1.26 (t , J = 7.1 Hz, 3H); ESI-MS (m/z): 251 (M+H) .
6
.3.4. Ethyl 4-(4-acetylphenoxy)butanoate (12):
–
1
white crystalline solid; yield: 90%; mp: 157-159 ºC; IR(KBr,cm ): 2935, 2352, 1675, 1566,
1
1
2
1
207, 1073, 770; H NMR (CDCl , 300 MHz) δ :7.93 (d, J = 8.7 Hz, 2H ) 6.92(d, J = 8.7 Hz,
3
H) 4.16 (q , J = 7.1 Hz, 2H) 4.09 (t , J = 6.1 Hz, 2H) 2.56-2.50 (m, 5H) 2.19-2.10 (m, 2H)
+
.27 (t, J = 7.1 Hz, 3H); ESI-MS (m/z): 251 (M+H) .
6
.4 General synthetic procedure for preparation of indole chalcone fibrates (13-30):
To a solution of appropriate indole-3-carbaldehyde (5-8) (1.0 eqiv.) in anhydrous MeOH,
substituted acetophenone (1.0 eqiv.) (9-12) and piperdine (0.01 eqiv.) were added. The
reaction mixture was refluxed for 24h. After completion of the reaction (monitored by TLC),
the reaction mixture was cooled to room temperature. The MeOH was evaporated in vacuum,
and the residue was diluted with water and the solution was extracted with CHCl (3 × 20
3
mL). The combined organic layers were dried over anhydrous Na SO and concentrated to
2
4
dryness to give a crude product that was purified by silica gel (100-200) column
chromatography (40% EtOAc/Hexane) to afford desired compounds (13-21) in moderate
yields. Similarly, compounds (22-30) were synthesised by using EtOH as solvent, in
moderate to good yields.
6
.4.1 (E)-methyl 2-(4-(3-(1H-indol-3-yl)acryloyl)phenoxy)propanoate (13):
–
1
Yellow solid, yield: 72%; mp: 125-127 ºC; IR (KBr, cm ): 3463, 3019, 1747, 1647, 1103,
1
7
61; H NMR (CDCl , 300 MHz) δ: 8.90 (s, 1H), 8.12-8.00 (m, 4H), 7.61-7.56 (m, 2H), 7.43
3
(s, 1H), 7.31-7.25 (m, 2H), 6.96 (d, J = 8.1 Hz, 2H), 4.88 (q, J = 6.7 Hz, 1H), 3.78 (s, 3H),
1
3
1
1
.67 (d, J = 6.7 Hz, 3H); C NMR (CDCl , 100 MHz) δ: 189.6, 172.3, 161.0, 138.8, 137.5,
3
32.7, 131.0, 130.7, 125.5, 123.4, 121.7, 120.7, 117.3, 114.8, 114.4, 112.3, 72.6, 52.6, 18.6;
+
ESI-MS (m/z): 350 (M+H) .
6
.4.2 (E)-methyl 2-(4-(3-(1H-indol-3-yl)acryloyl)phenoxy)butanoate (14):
–
1
Yellow solid, yield: 75%; mp: 141-143 ºC; IR (KBr, cm ): 3464, 3286, 1745, 1648, 1108,
1
7
61; H NMR (CDCl , 300 MHz) δ: 9.00 (s, 1H), 8.12-8.02 (m, 4H), 7.61-7.57 (m, 2H), 7.43
3
(s, 1H), 7.31-7.25 (m, 2H), 6.96 (d, J = 8.3 Hz, 2H), 4.7 (t, J = 6.0 Hz, 1H), 3.78 (s, 3H),
9

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1
3
2
1
1
.09-2.0 (m, 2H), 1.10 (t, J = 7.3 Hz, 3H); C NMR (CDCl , 100 MHz) δ: 189.6, 171.8,
3
61.3, 138.9, 137.5, 132.6, 131.0, 130.7, 125.4, 123.4, 121.6, 120.6, 117.1, 114.8, 114.2,
+
12.3, 77.6, 52.4, 26.1, 9.6; ESI-MS (m/z): 364 (M+H) .
6
.4.3 (E)-methyl 4-(4-(3-(1H-indol-3-yl)acryloyl)phenoxy)butanoate (15):
–
1
Yellow solid, yield: 78%; mp: 135-137 ºC; IR (KBr, cm ): 3387, 3020, 1732, 1648, 1104,
1
8
23; H NMR (CDCl , 300 MHz) δ: 9.02 (s, 1H), 8.13-8.05 (m, 4H), 7.63-7.58 (m, 2H), 7.44
3
(s, 1H), 7.31-7.30 (m, 2H), 6.97 (d, J = 8.2 Hz, 2H), 4.09 (t, J = 5.6 Hz, 2H), 3.70 (s, 3H),
1
3
2
1
1
.56 (t, J = 6.8 Hz, 2H), 2.20-2.11 (m, 2H); C NMR (CDCl +CD OD, 100 MHz) δ: 190.2,
3 3
73.9, 162.2, 139.4, 137.6, 131.6, 131.4, 130.3, 125.2, 122.8, 121.1, 120.1, 116.0, 114.0,
+
13.5, 112.1, 66.7, 51.3, 30.2, 24.2; ESI-MS (m/z): 364 (M+H) .
6
.4.4 (E)-methyl 4-(4-(3-(5-methoxy-1H-indol-3-yl)acryloyl)phenoxy)butanoate (16):
–
1
1
Yellow solid, yield: 80%; mp: 137-139 ºC; IR (KBr, cm ): 3280, 1729, 1641, 1170, 764; H
NMR (CDCl , 300 MHz) δ: 8.93 (s, 1H), 8.11-8.03 (m, 3H), 7.57-7.47 (m, 3H), 7.33 (d, J =
3
8
.7 Hz, 1H), 6.98-6.93 (m, 3H), 4.09 (t, J = 5.9 Hz, 2H), 3.91 (s, 3H), 3.70 (s, 3H), 2.55 (t, J
1
3
=
7.09 Hz, 2H), 2.19-2.11 (m, 2H); C NMR (CDCl +CD OD, 75 MHz) δ: 190.0, 173.9,
3 3
1
1
62.3, 155.3, 139.2, 132.5, 131.7, 131.3, 130.5, 126.2, 116.1, 114.2, 113.6, 112.8, 112.4,
+
03.2, 66.92, 56.0, 51.7, 30.5, 24.4; ESI-MS (m/z): 394 (M+H) .
6
.4.5 (E)-methyl 2-(4-(3-(5-chloro-1H-indol-3-yl)acryloyl)phenoxy)propanoate (17):
–
1
1
Yellow solid, yield: 78%; mp: 176-178 ºC; IR (KBr, cm ): 3287, 1748, 1654, 1154, 765; H
NMR (CDCl , 300 MHz) δ: 9.17 (s), 1H, 8.04-7.92 (m, 2H), 7.56-7.45 (m, 2H), 7.34 (d, J =
3
6
3
1
.4 Hz, 1H), 7.24-7.22 (m, 1H), 6.96 (d, J = 6.5 Hz, 2H), 4.89 (q, J = 5.0 Hz, 1H), 3.79 (s,
1
3
H), 1.67 (d, J = 5.0 Hz, 3H); C NMR (CDCl , 100 MHz) δ: 189.6, 172.3, 161.0, 138.8,
3
37.5, 132.7, 130.7, 125.5, 123.4, 121.7, 120.7, 114.8, 114.4, 112.3, 72.6, 52.6, 18.6; ESI-MS
+
(
m/z): 384 (M+H) .
6
.4.6 (E)-methyl 4-(4-(3-(5-chloro-1H-indol-3-yl)acryloyl)phenoxy)butanoate (18):
–
1
1
Yellow solid, yield: 82%; mp: 179-181 ºC; IR (KBr, cm ): 3190, 1731, 1647, 1169, 770; H
NMR (CDCl , 300 MHz) δ: 9.04 (s, 1H), 8.07-7.94 (m, 4H), 7.59 (d, J = 2.7 Hz, 1H), 7.51 (d,
3
J = 15.5 Hz, 1H), 7.36 (d, J = 8.6 Hz, 1H), 7.23 (d, J = 1.83 Hz, 1H), 6.98 (d, J = 8.7 Hz,
1
3
2
H), 4.10 (t, J = 6.1 Hz, 2H), 3.70 (s, 3H), 2.56 (t, J = 7.2 Hz, 2H), 2.20-2.21 (m, 2H); C
1
0

ACCEPTED MANUSCRIPT
NMR (CDCl +CD OD, 75 MHz) δ: 190.1, 173.9, 162.4, 138.6, 136.0, 132.3, 131.3, 130.5,
3
3
1
26.8,126.3, 123.1, 119.7, 116.5, 114.1, 113.1, 66.8, 51.5, 30.3, 24.3; ESI-MS (m/z): 398
+
(
M+H) .
6
.4.7 (E)-methyl 2-(4-(3-(2-methyl-1H-indol-3-yl)acryloyl)phenoxy)butanoate (19):
–
1
Yellow solid, yield: 82%; mp: 127-129 ºC; IR(KBr,cm ): 3456, 3020, 1745, 1644, 1172,
1
7
62; H NMR (CDCl , 300 MHz) δ: 8.83 (s, 1H), 8.15 (d, J = 15.3 Hz, 1H), 8.06 (d, J = 8.7
3
Hz, 2H), 7.93 (d, J = 7.4 Hz, 1H), 7.56 (d, J = 15.3 Hz, 1H), 7.35-7.32 (m, 1H), 7.28-7.19 (m,
H), 6.96 (d, J = 8.7 Hz, 2H), 4.69 (t, J = 6.1 Hz, 1H), 3.78 (s, 3H), 2.56 (s, 3H), 2.09-1.99
2
13
(
m, 2H), 1.10 (t, J = 7.4 Hz, 3H); C NMR (CDCl , 75 MHz) δ: 189.5, 171.8, 161.3, 142.7,
3
1
5
38.1, 136.2, 132.9, 130.6, 126.4, 122.4, 121.6, 120.2, 115.77, 114.8, 111.4, 110.6, 77.5,
+
2.4, 26.2, 12.3, 9.6; ESI-MS (m/z): 378 (M+H) .
6
.4.8 (E)-methyl 2-methyl-2-(4-(3-(2-methyl-1H-indol-3-yl)acryloyl)phenoxy)propanoate
(20):
–
1
Yellow solid, yield: 76%; mp: 132-134 ºC; IR(KBr,cm ): 3467, 3034, 1738, 1649, 1117,
1
7
2
2
67; H NMR (CDCl , 400 MHz) δ: 9.00 (s, 1H), 8.15 (d, J = 15.3 Hz, 1H), 8.04-8.02 (m,
3
H), 7.94 (d, J = 7.6 Hz, 1H), 7.56 (d, J = 15.3 Hz, 1H), 7.34-7.32 (m, 1H),7.24- 7.18 (m,
13
H), 6.92-6.88 (m, 2H), 3.78 (s, 1H), 2.54 (s, 3H), 1.68 (s, 6H); C NMR (CDCl , 100 MHz)
3
δ: 189.6, 174.6, 159.2, 142.6, 138.1, 136.2, 132.9, 130.2, 126.4, 122.6, 121.6, 120.2, 117.7,
+
1
15.8, 111.4, 110.7, 79.4, 52.8, 25.5, 12.4; ESI-MS (m/z): 378 (M+H) .
6
.4.9 (E)-methyl 4-(4-(3-(2-methyl-1H-indol-3-yl)acryloyl)phenoxy)butanoate (21):
–
1
Yellow solid, yield: 79%; mp: 137-139 ºC; IR (KBr, cm ): 3464, 3150, 1739, 1654, 1121,
1
7
86; H NMR (CDCl , 300 MHz) δ: 9.01 (s, 1H), 8.16 (d, J = 15.3 Hz, 1H), 8.06 (d, J = 8.4
3
Hz, 2H), 7.96 (d, J = 7.4 Hz, 1H), 7.60 (d, J = 15.3 Hz, 1H), 7.35 (d, J = 7.4 Hz, 1H), 7.29-
7
5
1
3
.19 (m, 2H), 6.98 (d, J = 8.4 Hz, 2H), 4.09 (t, J = 5.9 Hz, 2H), 3.70 (s, 3H), 2.59-2.55 (m,
1
3
H), 2.20-2.11 (m, 2H); C NMR (CDCl , 100 MHz) δ: 189.4, 173.7, 162.3, 142.1, 137.7,
3
36.1, 132.1, 130.6, 126.5, 122.5, 122.7, 121.6, 120.3, 116.2, 114.3, 111.3, 110.8, 67.0, 51.8,
+
0.6, 24.6, 12.5; ESI-MS (m/z): 378 (M+H) .
6
.4.10 (E)-ethyl 2-(4-(3-(1H-indol-3-yl)acryloyl)phenoxy)propanoate (22):
1
1

ACCEPTED MANUSCRIPT
–
1
Yellow solid, yield: 78%; mp: 129-131 ºC; IR (KBr, cm ): 3459, 3286, 1742, 1648, 1165,
1
7
7
61; H NMR (CDCl , 400 MHz) δ: 8.88 (s, 1H), 8.11-8.06 (m, 2H), 8.04-8.00 (m, 2H), 7.60-
3
.56 (m, 2H), 7.45-7.43 (m, 1H), 7.31-7.28 (m, 2H), 6.96 (d, J = 8.8 Hz, 2H), 4.85 (q, J = 6.8
1
3
Hz, 1H), 4.24 (q, J = 7.0 Hz, 2H), 1.67 (d, J = 6.6 Hz, 3H0, 1.26 (t, J = 7.1 Hz, 3H); C
NMR (CDCl , 100 MHz) δ: 189.8, 172.13, 161.2, 139.2, 137.7, 131.2, 130.8, 125.6, 123.6,
3
+
1
21.8, 120.8, 117.2, 114.9, 114.4, 112.5, 72.8, 61.9, 18.7, 14.4; ESI-MS (m/z): 364 (M+H) .
6
.4.11 (E)-ethyl 2-(4-(3-(1H-indol-3-yl)acryloyl)phenoxy)-2-methylpropanoate (23)
–
1
Yellow solid, yield: 85%; mp: 121-123 ºC; IR (KBr, cm ): 3461, 3283, 1731, 1646, 1173,
1
7
59; H NMR (CDCl , 300 MHz) δ: 8.97 (s, 1H), 8.12-8.01 (m, 4H), 7.62-7.58 (m, 2H), 7.44
3
(s, 1H), 7.31-7.30 (m, 2H), 6.90 (d, J = 8.1 Hz, 2H), 4.25 (q, J = 6.8 Hz, 2H), 1.68 (s, 6H),
1
3
1
1
1
.24 (t, J = 6.8 Hz, 3H); C NMR (CDCl , 75 MHz) δ: 189.7, 174.0, 159.3, 138.9, 137.5,
3
32.5, 130.8, 130.2, 125.4, 123.4, 121.7, 120.7, 117.7, 117.2, 114.3, 112.3, 79.4, 61.8, 25.5,
+
4.1; ESI-MS (m/z): 378 (M+H) .
6
.4.12 (E)-ethyl 4-(4-(3-(1H-indol-3-yl)acryloyl)phenoxy)butanoate (24)
–
1
Yellow solid, yield: 72%; mp: 119-121 ºC; IR (KBr, cm ): 3461, 3282, 1725, 1645, 1111,
1
7
59; H NMR (CDCl , 400 MHz) δ: 8.82 (s, 1H), 8.11-8.01 (m, 4H), 7.62-7.59 (m, 2H), 7.45
3
(m, 1H), 7.32-7.28 (m, 2H), 6.98 (d, J = 8.8 Hz, 2H), 4.16 (q, J = 7.1 Hz, 2H), 4.10 (t, J = 6.1
1
3
Hz, 2H), 2.54 (t, J = 7.2 Hz, 2H), 2.19-2.12 (m, 2H), 1.27 (t, J = 7.1 Hz, 3H); C NMR
(
DMSO-d +CD OD, 100 MHz) δ: 188.2,173.2, 162.6, 138.8, 138.0, 132.8, 131.8, 130.9,
6 3
1
25.7, 123.2, 121.6, 120.7, 115.9, 114.8, 113.4, 112.8, 67.4, 60.5, 30.6, 24.7, 14.3; ESI-MS
+
(
m/z): 378 (M+H) .
6
.4.13 (E)-ethyl 2-(4-(3-(5-methoxy-1H-indol-3-yl)acryloyl)phenoxy)propanoate (25):
–
1
Yellow solid, yield: 69%; mp: 131-133 ºC; IR (KBr, cm ): 3456, 3279, 1741, 1651, 1105,
1
7
7
4
84; H NMR (CDCl , 400 MHz) δ: 8.71 (s, 1H), 8.09-8.02 (m, 3H), 7.57 (d, J = 2.8 Hz, 1H),
3
.47 (d, J = 15.5 Hz, 1H), 7.42 (d, J= 2.4 Hz, 1H), 7.34-7.31 (m, 2H), 6.97-6.93 (m, 3H),
.85 (q, J= 6.8 Hz, 1H), 4.24 (q, J = 7.1 Hz, 2H), 3.91 (s, 3H), 1.66 (d, J = 6.8 Hz, 3H), 1.26
1
3
(
t, J = 7.1 Hz, 3H); C NMR (CDCl , 100 MHz) δ: 189.6, 171.8, 161.0, 155.6, 138.7, 132.6,
3
1
1
32.4, 130.7,130.6, 126.3, 116.9, 114.8, 114.1, 112.9, 112.8, 103.3, 72.6, 61.6, 56.1, 18.5,
+
4.2; ESI-MS (m/z): 394.2 (M+H) .
1
2

ACCEPTED MANUSCRIPT
6
.4.14 (E)-ethyl 2-(4-(3-(5-chloro-1H-indol-3-yl)acryloyl)phenoxy)propanoate (26):
–
1
Yellow solid, yield: 72%; mp: 183-185 ºC; IR (KBr,cm ): 3364, 3087, 1795, 1674, 1130,
1
7
7
2
7
1
94; H NMR (CDCl , 400 MHz) δ: 9.15 (s, 1H), 8.04-7.92 (m, 4H), 7.56 (d, J = 2.7 Hz, 1H),
3
.48 (d, J = 15.6 Hz, 1H), 7.34 (d, J = 8.6 Hz, 1H), 7.24-7.22 (m, 1H), 6.96 (d, J = 8.8 Hz,
H), 4.86 (q, J = 6.7 Hz, 1H), 4.25 (q, J = 7.1 Hz, 2H), 1.67 (d, J = 6.8 Hz, 3H), 1.27 (t, J =
13
.1 Hz, 3H); C NMR (CDCl , 100 MHz) δ: 189.9, 171.8, 160.8, 138.8, 135.8, 132.3, 131.8,
3
30.2, 126.6, 126.1, 122.8, 119.3, 116.0, 114.3, 112.9, 112.8, 72.0, 61.3, 17.7, 13.4; ESI-MS
+
(
m/z): 398 (M+H) .
6
.4.15 (E)-ethyl 2-(4-(3-(5-chloro-1H-indol-3-yl)acryloyl)phenoxy)butanoate (27):
–
1
Yellow solid, yield: 74%; mp: 182-184 ºC; IR(KBr,cm ): 3396, 3176, 1756, 1697, 1205,
1
7
7
2
3
1
2
98; H NMR (CDCl , 400 MHz) δ: 9.09 (s, 1H), 8.05-7.93 (m, 4H), 7.57 (d, J = 1.8 Hz, 1H),
3
.49 (d, J = 11.6 Hz, 1H), 7.35 (d, J = 6.4 Hz, 1H), 7.26-7.23 (m, 1H), 6.97 (d, J = 6.4 Hz,
H), 4.67 (t, J = 6.1 Hz, 1H), 4.25 (q, J = 7.0 Hz, 2H), 2.08-2.01 (m, 2H), 1.27 (t, J = 7.0 Hz,
13
H), 1.11 (t, J = 7.3 Hz, 3H); C NMR (CDCl , 75 MHz) δ: 189.6, 171.4, 161.5, 138.2,
3
35.9, 132.3, 131.8, 130.7, 127.3, 126.5, 123.6, 120.1, 117.4, 114.8, 113.7, 113.3, 77.6, 61.6,
+
6.1, 14.2, 9.6; ESI-MS (m/z): 412 (M+H) .
6
.4.16 (E)-ethyl 2-(4-(3-(2-methyl-1H-indol-3-yl)acryloyl)phenoxy)butanoate (28):
–
1
Yellow solid, yield: 74%; mp: 142-144 ºC; IR (KBr, cm ): 3389, 3256, 1759, 1681, 1131,
1
7
83; H NMR (CDCl , 400 MHz) δ: 8.67 (s, 1H), 8.15 (d, J = 15.3 Hz, 1H), 8.06 (d, J = 8.7
3
Hz, 2H), 7.95 (d, J = 7.5 Hz, 1H), 7.57 (d, J = 15.3 Hz, 1H), 7.35-7.26 (m, 2H), 7.24-7.20 (m,
1
2
H), 6.97 (d, J = 8.7 Hz, 2H), 4.66 (t, J= 6.1 Hz, 1H), 4.24 (q, J = 7.1 Hz, 2H), 2.58 (s, 3H),
13
.07-2.00 (m, 2H), 1.26 (t, J = 7.1 Hz, 3H), 1.11 (t, J = 7.4 Hz, 3H); C NMR (CDCl , 100
3
MHz) δ: 189.4, 171.4, 161.3, 142.6, 138.1, 136.1, 132.7, 126.4, 122.6, 121.6, 120.2, 115.8,
+
1
14.7, 111.4, 110.6, 77.7, 61.5, 26.2, 14.3, 12.4, 9.7; ESI-MS (m/z): 392 (M+H) .
6
.4.17 (E)-ethyl 2-methyl-2-(4-(3-(2-methyl-1H-indol-3-yl)acryloyl)phenoxy)propanoate
(29):
–
1
Yellow solid, yield: 71%; mp: 137-139 ºC; IR (KBr, cm ): 3489, 3179, 1759, 1639, 1123,
1
7
71; H NMR (CDCl , 400 MHz) δ: 8.67 (s, 1H), 8.15 (d, J = 15.3 Hz, 1H), 8.03 (d, J = 8.8
3
Hz, 2H), 7.95 (d, J = 7.6 Hz, 1H), 7.57 (d, J = 15.3 Hz, 1H), 7.35-7.28 (m, 2H), 7.27-7.20 (m,
1
3

ACCEPTED MANUSCRIPT
2
7
1
H), 6.90 (d, J = 8.8 Hz, 2H), 4.25 (q, J = 7.1 Hz, 2H), 2.57 (s, 3H), 1.67 (s, 6H), 1.24 (t, J =
1
3
.1 Hz, 3H); C NMR (CDCl , 75 MHz) δ: 189.7, 174.0, 159.3, 142.7, 138.2, 136.2, 132.7,
3
30.2, 126.4, 122.7, 121.7, 120.2, 117.7, 115.7, 111.4, 110.6, 79.4, 61.8, 25.5, 14.1, 12.4;
+
ESI-MS (m/z): 392 (M+H) .
6
.4.18 (E)-ethyl 4-(4-(3-(2-methyl-1H-indol-3-yl)acryloyl)phenoxy)butanoate (30):
–
1
Yellow solid, yield: 78%; mp: 135-137 ºC; IR (KBr, cm ): 3440, 3296, 1789, 1656, 1134,
1
7
97; H NMR (CDCl , 300 MHz) δ: 8.82 (s, 1H), 8.16 (d, J = 15.4 Hz, 1H), 8.08 (d, J = 8.3
3
Hz, 2H), 7.96 (d, J = 7.2 Hz, 1H), 7.59 (d, J = 15.4 Hz, 1H), 7.35 (d, J = 7.2 Hz, 1H), 7.29-
.21 (m, 2H), 6.98 (d, J = 8.4 Hz, 2H), 4.19-4.08 (m, 4H), 2.57-2.52 (m, 5H), 2.15 (t, J = 6.4
7
1
3
Hz, 2H), 1.29-1.24 (m, 3H); C NMR (CDCl +CD OD, 75 MHz) δ: 190.1, 173.5, 162.3,
3
3
1
3
43.3, 138.5, 136.3, 132.0, 126.2, 122.3, 121.3, 120.0, 115.0, 114.2, 11.3, 110.2, 66.9, 60.6,
+
0.7, 24.4, 14.0, 11.8; ESI-MS (m/z): 392 (M+H) .
Acknowledgements
Research work is supported by CSIR-CDRI Network project: "Towards holistic
understanding of complex diseases: Unraveling the threads of complex disease (THUNDER)
Project No: BSC0102. We wish to thank Dr.S.P. Singh for technical support, SAIF for NMR,
IR, and Mass spectral data. R. P. D., G.R.P. and R.S. are thankful to CSIR, New Delhi, India
for financial support. This is CSIR-CDRI communication number 69/2014/KVS.
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Captions to illustrations.
Table 1. The lipid lowering activity of indole-chalcone fibrates (100 mg/kg) in triton induced
hyperlipidemic rats. Triton treated group is compared with control and drug treated group is
compared with triton group.
Figure 1. Chemical structures of some representative indoles, fibrates and chalcones
possessing lipid lowering activities.
Figure 2. Rational designing and preliminary SAR of indole-chalcone fibrates.
Figure 3. Percentage (%) change of plasma lipids with the treatment of compounds 19, 24
and 29 at different doses (50, 100 & 150mg/kg b.wt) in triton-induced hyperlipidemic rats at
the different doses. Each parameter represents pooled data from 6 rats/group and values are
NS
expressed as mean±S.D. ***P < 0.001; non significant, comparison b/w compounds treated
different doses. (50,100 and 150 mg/kg b.wt).
Figure 4. Effect of compounds 19, 24 and 29 on lipoprotein metabolism in triton induced
hyperlipidemic rats. Each parameter represents pooled data from 6 rats/group and values are
expressed as mean±S.D. ***P < 0.001 compared between control and triton treated rats group
only, bP < 0.01; cP < 0.001between triton and triton plus compounds treated groups,
fenofibrate was taken as standard drug.
Figure 5. Compounds 19, 24 and 29 at dose of 100mg/kg p.o re-activate LPL activity in
triton induced hyperlipidemic rats. Each parameter represents pooled data from 6 rats/group
and values are expressed as mean±S.D. ***P < 0.001 between control and triton treated rats
group only, aP < 0.05; bP < 0.01 between triton and triton plus compounds treated groups,
fenofibrate was taken as standard drug.
Figure 6. The effect of indole-chalcone fibrate compounds (200ꢀg/ml) on superoxide ions
(nmol.formazone formed/min), hydroxyl ion (nmol. MDA formed/h) and lipid peroxidation
in microsomes (nmol. MDA formed/mg protein) was shown (standard drugs for superoxide
anions-alloperinol (200ꢀg/ml), hydroxyl ions-manitol and for microsomal lipid peroxidation-
α-tocopherol (200ꢀg/ml) were used. Each value is mean±SD of six values, *P < 0.05; **P <
0
.01; ***P < 0.001 experimental values compared with control values.
Scheme 1. Synthesis of novel indole-chalcone fibrates. Reagents and conditions: (a) POCl3,
o
o
DMF, 0 C, 20% aq NaOH, reflux, 6h; (b) K CO , CH CN, 100 C, 4h; (c) piperdine, MeOH,
2
3
3
2
0h; (d) piperdine, EtOH, 20h.
2
0

ACCEPTED MANUSCRIPT
Table 1. Percentage (%) change of plasma lipids with the treatment of indole-chalcone
fibrates (13-30) in triton-induced hyperlipidemic rats at the dose of 100 mg/kg body weight.
Compound
no.
Lipid profile
Phospholipids Triglyceride
b
Structure
Total cholesterol
PHLA
a
a
a
(TC)
(PL)
(TG)
O
O
O
O
1
1
3
4
NS
-
13*
-14*
-12*
+6
+3
N
H
O
O
O
NS
NS
NS
NS
O
-8
-8
-9
N
H
O
O
O
O
1
1
5
6
-20**
-22**
-11*
-22**
-14*
+14*
N
H
O
O
O
O
O
NS
-13*
+7
N
H
O
O
O
Cl
1
1
7
8
NS
O
-
12*
10*
-14*
-11*
-17*
-10*
+3
N
H
O
O
Cl
O
NS
-
+4
O
N
H
O
O
O
O
1
2
9
0
-
26***
-24***
-12*
-27***
-10*
+16*
N
H
O
O
O
NS
O
-10*
+6
N
H
O
O
O
2
1
NS
-
15*
-18*
-16*
-12*
+9
O
N
H
O
O
O
O
2
2
2
3
NS
-15*
-13*
+8
N
H
O
O
O
O
-20**
-22***
-22***
+14*
N
H
2
1

ACCEPTED MANUSCRIPT
O
O
O
2
4
O
-31***
-32***
-13*
-30***
-10*
+19*
N
H
O
O
O
O
2
5
6
O
NS
-11*
+6
N
H
O
O
O
O
Cl
2
NS
-
11*
18*
-15*
-16*
-13*
-16*
+7
N
H
O
O
O
O
Cl
2
7
-
+13*
N
H
O
O
O
O
2
2
8
9
-
20**
-22**
-33***
-16*
-22**
-30***
-14*
+14*
N
H
O
O
O
O
-
32***
+20**
N
H
3
0
NS
-13*
+7
…
…
Fenofibrate
-32***
-30***
-33***
+19*
a
b
:
mg/dl; : n mol of free fatty acids formed/h/ml of plasma; Values are mean ± SD of six rats, ***p <0.001; **p
<
0.01; *p <0.05; NS = Non significant. The lipid lowering activity of indole-chalcone fibrates (100 mg/kg) in
triton induced hyperlipidemic rats. Triton treated group is compared with control and drug treated group is
compared with triton group.
2
2

ACCEPTED MANUSCRIPT
Figure 1. Chemical structures of some representative indoles, fibrates and chalcones
possessing lipid lowering activities.
2
3

ACCEPTED MANUSCRIPT
Figure 2. Rational designing and preliminary SAR of indole-chalcone fibrates.
2
4

ACCEPTED MANUSCRIPT
Figure 3. Percentage (%) change of plasma lipids with the treatment of compounds 19, 24
and 29 at different doses (50, 100 & 150mg/kg b.wt) in triton-induced hyperlipidemic rats at
the different doses. Each parameter represents pooled data from 6 rats/group and values are
NS
expressed as mean±S.D. ***P < 0.001; non significant, comparison b/w compounds treated
different doses. (50,100 and 150 mg/kg b.wt)
2
5

ACCEPTED MANUSCRIPT
Figure 4. Effect of compounds 19, 24 and 29 on lipoprotein metabolism in triton induced
hyperlipidemic rats. Each parameter represents pooled data from 6 rats/group and values are
expressed as mean±S.D. ***P < 0.001 compared between control and triton treated rats group
b
c
only, P < 0.01; P < 0.001between triton and triton plus compounds treated groups,
fenofibrate was taken as standard drug.
2
6

ACCEPTED MANUSCRIPT
Figure 5. Compounds 19, 24 and 29 at dose of 100mg/kg p.o re-activate LPL activity in
triton induced hyperlipidemic rats. Each parameter represents pooled data from 6 rats/group
and values are expressed as mean±S.D. ***P < 0.001 between control and triton treated rats
a
b
group only, P < 0.05; P < 0.01 between triton and triton plus compounds treated groups,
fenofibrate was taken as standard drug.
2
7

ACCEPTED MANUSCRIPT
Figure 6. The effect of indole-chalcone fibrate Compounds (200ꢀg/ml) on superoxide ions
(nmol.formazone formed/min), hydroxyl ion (nmol. MDA formed/h) and lipid peroxidation
in microsomes (nmol. MDA formed/mg protein) was shown (standard drugs for superoxide
anions-alloperinol (200ꢀg/ml), hydroxyl ions-manitol and for microsomal lipid peroxidation-
α-tocopherol (200ꢀg/ml) were used. Each value is mean±SD of six values, *P < 0.05; **P <
0
.01; ***P < 0.001 experimental values compared with control values.
2
8