Synthesis of N-(5-chloro-6-(quinolin-3-yloxy)pyridin-3-yl) benzenesulfonamide derivatives as non-TZD peroxisome proliferator-activated receptor γ (PPARγ) agonist

Article abstract of DOI:10.1016/j.ejmech.2012.10.027

The thiazolidinediones (TZDs) are a class of oral antidiabetic drugs that improve insulin sensitivity in patients with type 2 diabetes. Although the mechanism by which the TZDs lower insulin resistance is unclear, they are known to target the peroxisome p

Full text of DOI:10.1016/j.ejmech.2012.10.027

Accepted Manuscript
Synthesis of N-(5-chloro-6-(quinolin-3-yloxy)pyridin-3-yl) benzenesulfonamide
derivatives as Non-TZD Peroxisome Proliferator-Activated Receptor γ (PPARγ)
agonist
Swapnil Bajare, Jessy Anthony, Amrutha Nair, Rosalind Marita, Anagha Damre,
Dharmeshkumar Patel, Chandrika Rao, H. Sivaramakrishnan, Nabajyoti Deka
PII:
S0223-5234(12)00631-9
10.1016/j.ejmech.2012.10.027
EJMECH 5796
DOI:
Reference:
To appear in: European Journal of Medicinal Chemistry
Received Date: 23 July 2012
Revised Date: 9 October 2012
Accepted Date: 16 October 2012
Please cite this article as: S. Bajare, J. Anthony, A. Nair, R. Marita, A. Damre, D. Patel, C.
Rao, H Sivaramakrishnan, N. Deka, Synthesis of N-(5-chloro-6-(quinolin-3-yloxy)pyridin-3-yl)
benzenesulfonamide derivatives as Non-TZD Peroxisome Proliferator-Activated Receptor γ (PPARγ)
agonist, European Journal of Medicinal Chemistry (2012), doi: 10.1016/j.ejmech.2012.10.027.
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Graphical Abstract:

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Research Highlight
ꢀ N-(5-chloro-6-(quinolin-3-yloxy)pyridin-3-yl)benzenesulfonamides were
synthesized
ꢀ Synthesized compounds exhibited adipogenesis activity
ꢀ PPARγ agonism of the series translated into in vivo efficacy in db/db mice.

ACCEPTED MANUSCRIPT
Synthesis of N-(5-chloro-6-(quinolin-3-yloxy)pyridin-3-yl) benzenesulfonamide derivatives as
Non-TZD Peroxisome Proliferator-Activated Receptor γ (PPARγ) agonist
Swapnil Bajare,a Jessy Anthony,b Amrutha Nair,b Rosalind Marita,b Anagha Damre,d Dharmeshkumar
Patel,c Chandrika Rao,c H Sivaramakrishnan,a and Nabajyoti Deka*a
Piramal Healthcare Limited, Goregaon (E), Mumbai 400 063, Maharashtra, India
* - To whom correspondence should be addressed, Address: 1 Nirlon Complex, Off Western Express
Highway, Goregaon (East), Mumbai-400063, India; Phone: +91-22-30818320; Fax: +91-22-30818036;
e-mail: nabajyoti.deka@piramal.com
a - Department of Medicinal Chemistry, Piramal Healthcare Limited
b - Department of Pharmacology, Piramal Healthcare Limited
c - Department of Discovery Informatics, Piramal Healthcare Limited
d – Department of DMPK, Piramal Healthcare Limited
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Abstract
The thiazolidinediones (TZDs) are a class of oral antidiabetic drugs that improve insulin sensitivity in
patients with type 2 diabetes. Although the mechanism by which the TZDs lower insulin resistance is
unclear, they are known to target the peroxisome proliferator–activated receptor-γ (PPARγ)-a nuclear
hormone receptor. Ligands for PPARγ regulate adipocyte production and secretion of fatty acids as
well as glucose metabolism, resulting in increased insulin sensitivity in adipose tissue, liver, and
skeletal muscle. However, TZDs have several adverse effects, including weight gain and liver toxicity.
Herein we report identification of non-TZD PPARγ agonists which exhibits beneficial effects similar to
that of TZDs in animal models, but without the associated adverse effects.
Key Words: Thiazolidinediones; adipocyte; PPARγ; type 2 diabetes mellitus; adipogenesis
Introduction
The Thiazolidinediones (TZDs) class of drugs improves insulin sensitivity [1], glucose tolerance, and
lipid homeostasis in vivo. TZDs decrease circulating levels of insulin [2], free fatty acids, and
triglycerides and increase insulin-stimulated glucose uptake and utilization by activating the nuclear
receptor PPARγ [3, 4]. A potential mechanism for these beneficial metabolic effects is the net
partitioning of fatty acids from elsewhere in the body to adipose tissue for storage [5]. They bind to
PPARγ with high affinity and specificity, promote interaction of PPARγ with transcriptional
coactivators or corepressors, increase PPARγ mediated transcription regulation, and promote PPARγ
mediated cellular effects such as adipogenesis. PPARγ and TZDs regulate the expression of several
dozens of genes involved in a variety of cellular functions [6], including the metabolism of
carbohydrates, fatty acids, triglycerides, and cholesterol. It is not clear which, if any, of these genes
may be critical for insulin sensitization. It is also unknown which of the insulin-responsive tissues is
the key target of TZDs, though adipose tissue is the leading candidate [7]. PPARγ is expressed at high
levels in the adipose tissue but at much lower levels in muscle and liver tissues. PPARγ agonists
regulate adipocyte differentiation and metabolism [8].
During the development of PPARγ agonists, many of these agents have been plagued with safety
issues. There are few TZDs that have come to clinical use - troglitazone, rosiglitazone, and
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pioglitazone (Figure-1) as well as several others that have been limited to pre-clinical study. The first
TZD PPARγ agonist for type 2 diabetes, troglitazone was launched in the US in 1997 and withdrawn in
2000 due to serious hepatotoxicity [9] seen in a small number of patients during the postmarketing
period. The currently approved PPARγ agonists, pioglitazone and rosiglitazone, do not display this
hepatotoxicity. However, since weight gain and edema have been reported as side effects of these
drugs, improvement of PPARγ agonists as antidiabetic agents is still required. INT-131 (Figure-1), a
novel, non-thiazolidinedione (TZD), selective peroxisome proliferator-activated receptor gamma
(PPARγ) modulator, is in development by InteKrin Therapeutics Inc for the treatment of type 2
diabetes mellitus (non-insulin dependent diabetes) [10]. In our current research program towards
development of non-TZD PPARγ agonist to overcome the well-known shortcomings of
thiazolidinediones, we have made efforts to synthesize ligand for PPARγ that is structurally and
pharmacologically distinct from glitazone agonists. The concept of selective modulation involves
targeting and activating specific genes to minimize side effects while maintaining therapeutic benefits.
Modification of the structural name leads and their effects on in vitro measures of adipogenesis,
hPPARγ agonism and in vivo efficacy in db/db mice are disclosed in this communication.
In this connection with the attempt of developing non-TZD selective PPARγ ligands, we have designed
and synthesized N-(5-chloro-6-(quinolin-3-yloxy)pyridin-3-yl)benzenesulfonamide derivatives based
on the chemical structure of INT-131 and screen them for adipogenesis in 3T3-1 adipocytes and
PPARγ transactivation activity. The nuclear receptor, PPARγ, is the master regulator of adipogenesis
and PPARγ active compounds exhibits adipogenic activity. Since the adipogenesis assay is an intricate
phenotype-based assay, a direct structure activity relationship may not be imperative.
In vitro assays showed that N-(5-chloro-6-(quinolin-3-yloxy)pyridin-3-yl)benzenesulfonamide
derivatives activate PPARγ. In adipocytes, they showed stimulation of adipocyte differentiation
indicating that these compounds may activate PPARγ target genes involved in adipogenesis. The
synthesis
of
novel
non
TZD
ligands,
N-(5-chloro-6-(quinolin-3-yloxy)pyridin-3-
yl)benzenesulfonamide derivatives (7-20) is depicted in scheme-1.
Synthesis of the target componds involved straightforward chemical transformations and
commenced with the preparation of 2-hydroxy-3-chloro-5-nitropyridine (2). Commercially
available 2-hydroxy-5-nitropyridine (1) was treated with concentrated HCl at 50 ºC followed by
aqueous solution of sodium chlorate to afforded 2-hydroxy-3-chloro-5-nitropyridine (2) in
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quantitative yield [11]. Compound 2 was converted to required intermediate 2,3-dichloro-5-
nitropyridine (3) by using POCl3 at 120 ºC. Coupling reaction of 2,3-dichloro-5-nitropyridine (3)
and Quinolin-3-ol (4) was carried out in presence of Cs2CO3 in DMF. Other reagents like
K2CO3/THF also used to afford 3-((3-chloro-5-nitropyridin-2-yl)oxy)quinoline (5) in quantitative
yield. Subsequent reduction of the compound 5 was carried out by using SnCl2.H2O in ethyl
acetate. Target compounds (5-chloro-6-(quinolin-3-yloxy)pyridin-3-yl)benzenesulfonamide
derivatives (7-20) were synthesized by treating the amine 6 with different sulfonyl chlorides in
DCM / pyridine at r.t. Purifications were carried out by either coloumn chromatography or by
recrystallization.
The synthesized compounds exhibited stimulation of adipocyte differentiation and activate PPARγ
which indicated that these compounds may activate PPARγ target genes involved in adipogenesis.
A representative compound 7 was docked and compared with INT131 and exhibited comparable
similarities and interactions in the active site of PPARγ pocket.
Compound 7 superposes very well on INT131 and Both the compounds hug the H3 helix with
hydrophobic interactions and form water-mediated H-bonds with AF2 helix (Figure-2).
Figure-3 shows that N-atom of pyridine ring in compound 7 is differently placed in a hydrophobic
region of the active site. This may have additional effects on the activity of the molecule. INT131
wraps around the H3 helix without getting close to the AF2 helix. While direct interaction with
AF2 helix plays a major role in the binding affinity of PPARγ agonists, INT131 has water-
mediated interactions with AF2 helix [12], which may reduce the strength of binding. 2,4 dichloro
benzene of INT131 has dense hydrophobic interactions with hydrophobic cavity of Phe360,
Phe363, Ile281, Phe282, Cys285,Leu356. In compound 7 dichlorobenzene is replaced by
dimethoxybenzene. While hydrophobic interactions of aromatic ring are maintained as in INT131,
interactions of Cl are lost and oxygen atoms have no hydrophobic interactions.
Gold Score for compound 7 is 90.65 which is equivalent to that of INT131 while Hydrogen Bonding
Score is 6.8 for compound 7 and 7.6 for INT131. These docking data indicated that compound 7 and
INT131 exhibited comparable affinity for PPARγ and similar ligand protein interactions.
All new compounds were evaluated as ligands for the human PPARγ, in HEK -293 cell-based
PPARγ transactivation assay for agonist activity . Active compounds were also evaluated for their
in vitro adipogenic potential in 3T3-L1 adipocytes. In vitro profiling of the compounds have been
summarized in detail in Table 1 .
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Following the identification of compounds with good intrinsic potency in vitro, it was necessary to
determine whether these compounds would retain useful in vivo efficacy in diabetes models while
exhibiting an enhanced safety profile with respect to known PPARγ mechanism-based liabilities. Easily
monitored liabilities include weight gain. In vivo testing in rodent models was used to ascertain
whether this objective could be attained. Db/db mice, a genetic model of obese, insulin resistant
T2DM, was used to evaluate the in vivo efficacy of analogs with adequate pharmacokinetic profiles.
Compound 7 showed a Cmax of 33.9 µg/mL at 0.25 h. The area under the plasma concentration-time
curve (AUC0-8h) was 146.44 µg*h/mL and the plasma half-life was 2.99 h. Based on the in vitro
activity and pharmacokinetic profile (Table 2), we selected compound 7 and 16 for further evaluation.
A number of compounds were tested in the db/db mouse model and efficacy data of the representative
compound 7 is described in detail.
Compound 7 effectively reduces hyperglycemia in the db/db model after a treatment of 50
mg/kg/b.i.d for 10 days (40 % plasma glucose reduction) . In the same model the known anti
diabetic agent ,rosiglitazone was effective as well in its glucose reduction activity (5 mg/kg/b.i.d,
52% glucose reduction).
Body weight gain measured in the db/db mouse amounted to only 9% at even at a high dose of 50
mg/kg/b.i.d of compound 7, while a 14 % body weight gain was noted with rosiglitazone at a dose
of 5 mg/kg/b.i.d. Therefore the efficacy study with compound 7 displayed significant glucose
lowering activity (Figure-4) and a reduced effect on body weight gain as compared to
rosiglitazone (Figure-5) indicating that compound 7, a non-TZD PPARγ agonist possesses
important pharmacological advantages relative to the TZD PPARγ agonist rosiglitazone in this
animal model.
Conclusion
Here in this study we report design and synthesis of N-(5-chloro-6-(quinolin-3-yloxy)pyridin-3-yl)
benzene-sulfonamide derivatives as novel non TZD PPARγ agonist for the develpement of safer
antidiabetic agent. The synthesized molecules exhibited adipogenesis activity and PPARγ agonism
leading to anti-diabetic effect in db/db mice. In comparison with the PPARγ full agonist
Rosiglitazone, adverse effects such as body weight gain was attenuated. Further investigations to
enhance these desirable profiles are ongoing.
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Acknowledgements
We thank the Department of Analytical Chemistry for providing us with NMR, mass, HRMS and
HPLC data.
Experimentals
All reagents and solvents were obtained from commercial sources and used as received.
1
13
H-NMR and C-NMR spectra were obtained on a ‘Bruker 300 MHz’ instrument equipped with a 5 mm
1H/13C/X (BBO) probe and the solvent indicated with tetramethylsilane as an internal standard. The data
obtained so, were processed and analyzed by using Bruker software, XWIN NMR version 3.5.
Analytical HPLC was run using a Zorbax Eclipse XDB-C8 3.5 ꢀm 4.6x75 mm column eluting with a
mixture of acetonitrile and water containing 0.1% trifluoroacetic acid with a 5 minute gradient of 10-100%.
MS results were obtained on ‘ESI-QTOF’ instruments of Bruker Daltonics (model MicrotofQ). 10 µl of
each sample (fraction) was injected. The sample was ionized using Electron Spray Ionisation technique and
analyzed using Quadruple Time of Flight. The mobile phase used was Acetonitrile and of 0.1 % formic
acid (50:50) with a flow rate of 0.2 ml/min. The samples were analyzed both in the positive mode and
negative mode by Direct Injection Mode.
Liquid chromatography/ mass spectroscopy studies have been carried out using ‘Agilent 1100 Series/
esquire 4000’ instrument of Bruker Daltonics. Same analytical HPLC method was used with ‘Phenomenex
Luna C18’ column. 0.1% Formic Acid and Acetonitrile (80:20) were used as mobile phase. 10µl of sample
was injected on ESI source. Ion source parameters were employed as required. Nebulizer pressure = 45 psi,
Dry Gas = 12 Lit/min, Dry Temp = 3500C, scan = 50 to 2200, Capillary Voltage = 4500, Polarity was
checked for both positive and negative mode. The data obtained so, were processed and analyzed by using
Hystar software.
Automated column chromatography was performed on a CombiFlash Rf 200 (Teledyne Isco Inc.). Melting
points were taken on a Mel-Temp apparatus and are uncorrected.
The protein structure of PPARg (PDB ID-3FUR), which was used for docking, was downloaded from
ProteinData Bank (www.rcsb.org/pdb). This is a co-crystal structure of PPARg with INT131. Compounds
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were docked to the binding site by means of CCDC’s GOLD (Genetic Optimization for Ligand Docking)
software, version 5.1. The binding region for the docking study was defined as all atoms within 6Å radius
sphere centered on the centroid of the INT131. Thirty genetic algorithm (GA) runs were performed with
automatic settings for each compound. The scoring function, GoldScore, implemented in GOLD was used to
rank the docking positions of compound.
The compounds were tested for adipogenic activity in the presence of insulin in 3T3-L1 cells at a
concentration of 20µg/ml. The adipogenic activity in the presence of potent PPARγ full agonist (1 µM) was
defined as 100%, the maximum adipogenic activity in the presence of the test compound was defined as Emax
(%). The compounds were tested for agonist activity on PPAR in transiently transfected HEK 293 cells at a
concentration of 10µM. The transcriptional activity in the presence of potent PPARγ full agonist (1µM) was
defined as 100%, the maximum transcriptional activity in the presence of the test compound was defined as
Emax (%).
Pharmacokinetic parameters were assessed following oral dosing (100 mg/kg) using a suspension
formulation (using 0.5% CMC and Tween 80; dosing volume: 10 mL/kg). Female db/db mice were
weighed and the compounds were administered orally (n = 4 per time point). Blood samples were
withdrawn at 0.08, 0.25, 0.5, 0.75, 1.0, 2.0, 6.0 and 8.0 h after dosing. Plasma samples were maintained
on ice before being centrifuged (4 ºC for 5 min at 1411g), and aliquots were stored at -80 ºC pending
the assay. Concentrations of the compounds were determined using an HPLC method developed at
Piramal Healthcare Limited. Pharmacokinetic parameters were determined by non-compartmental
analysis using WinNonlin Professional (Version 4.1). Cmax and Tmax were taken directly from the
plasma concentration-time profile. The area under the curve from time 0 to the last blood sampling
time (AUC0-t) was calculated using the linear trapezoidal rule. The area under the curve extrapolated to
infinity (AUCinf) was calculated by using the plasma concentration at time t divided by slope λz, where
λz is estimated by linear regression of the terminal log-linear phase of the plasma concentration-time
curve. Terminal plasma elimination half life (T1/2) was calculated as 0.693/ λz.
All animal experiments were performed according to procedures approved by the CPCSEA and as per the
IAEC guidelines. In brief, 5 to 7 week-old male db/db mice, bred at Piramal Healthcare Ltd were fed a chow
diet. The mice were treated with the respective compounds, body weight and biochemical parameters were
evaluated at the end of the study.
Analytical Data:
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2-Hydroxy-3-chloro-5-nitro pyridine (2).
2-Hydroxy-5-nitro pyridine (200 g, 1.42 mol) was added portion wise to 800 ml of concentrated HCl under
constant stirring and then heated to 50o C. To this was slowly added a solution of sodium chlorate (80.0 g,
0.75 mol) in water. The reaction was maintained at the same temperature for an additional hour, and then
cooled to 0o C. The precipitate obtained was filtered, washed thoroughly with water and dried to give 2-
Hydroxy-3-chloro-5-nitro pyridine. Yield: 240 g, 1.37 mmol (96.0%)
M.P.: 195° - 197°C
1H NMR (300 MHz, DMSO-d6): δ 8.65 (d, 1H, J = 2.5 Hz), 8.36 (d, 1H, J = 2.5 Hz).
13C NMR (300 MHz, DMSO-d6): δ 153.40, 144.29, 143.59, 135.08, 130.44
MS (ES): 175 (M+H)+
2, 3-Dichloro-5-nitro pyridine (3).
Quinoline (48.5 ml, 0.41 mol) was added to POCl3 (164.0 ml 1.75 mol) at 0o C under nitrogen. To this
stirred mixture, 2-hydroxy-3-chloro-5-nitro pyridine (204 mg, 1.17 mol) was added. The reaction mixture
was heated at 120o C for 2 hours, cooled to 0o C followed by addition of ice-cold water. The precipitate
obtained was filtered, washed thoroughly with water and dried to give 2,3-Dichloro-5-nitro pyridine.
Yield: 220 g, (97%)
M.P.: 53° C.
1H NMR (300 MHz, DMSO-d6): δ 9.16 (d, 1H, J = 2.4 Hz), 8.94 (d, 1H, J = 2.4 Hz).
13C NMR (300 MHz, DMSO-d6): δ 158.69, 137.36, 132.51, 130.13, 124.39
MS (ES): 193 (M+1)
3-((3-chloro-5-nitropyridin-2-yl)oxy)quinoline (5)
In a 250 ml of round bottom flask, 14.5 g (0.1 mol) of quinolin-3-ol was placed and 100 ml of dry dimethyl
formamide was added. To the stirred solution, 39.0 g (0.12 mol) of Cs2CO3 was added at 0 0C. Stirring was
continued and after 30 minutes 1.9 g (0.1 mol) of 2,3-dichloro-5-nitro pyridine (3) was added. Stirring was
continued further for 3-4 hours and the reaction was monitored by TLC. After completion, the reaction
mixture was poured into ice-water and extracted with ethyl acetate. Organic layer was separated and dried
over sodium sulfate (Na2SO4) solvent was removed under vacuum and to the resulting mass was added
water (50 ml), extracted with ethyl acetate, dried over sodium sulfate and concentrated under vacuum to
obtain crude 3-((3-chloro-5-nitropyridin-2-yl)oxy)quinoline that was purified using flash chromatography.
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1H NMR (300 MHz, DMSO-d6): δ 9.03 (d, J=2.4Hz, 2H), 8.95 (d, J=2.4Hz, 1H), 8.35 (d, J=2.7Hz, 1H),
8.09 (d, J=8.1Hz, 1H), 7.98 (d, J=8.1Hz, 1H), 7.82 (t, J=6.9Hz, 1H), 7.69 (t, J=8.1Hz, 1H).
HRMS (m/z): [M]+ calcd for C14H8ClN3O3, 301.0323; found, 301.1109
5-chloro-6-(quinolin-3-yloxy)pyridin-3-amine (6):
In a 250 ml of round bottom flask, 7.5 g (0.025 mol) of 3-((3-chloro-5-nitropyridin-2-yl)oxy)quinoline was
placed and 100 ml of ethyl acetate was added. To the stirred solution 220 g (0.10 mol) of SnCl2.2H2O was
added at room temperature and stirring was continued for 8 hours. After completion (monitored by TLC),
evaporated the solvent and added 25 ml of water followed by 1 N NaOH solution (pH was maintained
between 9 and 10). Ethyl acetate (50 ml x 3) was used for extraction. The combined ethyl acetate layers
was dried over sodium sulfate and concentrated under reduced pressure. The crude compound obtained
was purified by column chromatography using 30% ethyl acetate in petroleum ether solvent system which
gave 4.75 g (0.175 mol) of 5-chloro-6-(quinolin-3-yloxy)pyridin-3-amine. Yield: 70%
1H NMR (300 MHz, DMSO-d6): δ 8.75 (d, J=2.7Hz, 1H), 8.01 (d, J=2.7Hz, 1H), 7.92 (dd, J=7.5, 1.2Hz,
1H), 7.85 (d, J=2.7Hz, 1H), 7.70 (td, J=6.9, 1.5Hz, 1H), 7.60 (td, J=8.0, 1.2Hz, 1H), 7.49 (d, J=2.4Hz, 1H),
7.26 (d, J=2.4Hz, 1H), 5.49 (s, 2H)
HRMS (m/z): [M]+ calcd for C14H10ClN3O, 271.0511; found, 271.6109
General
Procedure
for
the
synthesis
of
N-(5-chloro-6-(quinolin-3-yloxy)pyridin-3-
yl)benzenesulfonamide (7-19) derivatives.
868 mg (3.2 mmol) of the amine, 5-chloro-6-(quinolin-3-yloxy)pyridin-3-amine was dissolve in dry DCM
and 2 ml of pyridine was added. To the stirred solution, 3.2 mmol of desired sulfonyl chloride was added.
The reaction mixture was stirred at room temperature 5-6 hours. After completion, evaporated the solvent
and the resulting crude product was purified using ethyl acetate and pet-ether solvent system. All purified
compounds (7-15) were characterized by spectral analysis
N-(5-chloro-6-(quinolin-3-yloxy)pyridin-3-yl)-3,4-dimethoxybenzenesulfonamide (7).
1H NMR (300 MHz, DMSO-d6): δ 10.40 (s, 1H), 8.77 (d, J=2.4Hz, 1H), 8.14 (d, J=2.1Hz, 1H), 8.04 (d,
J=7.8Hz, 1H), 7.94 (d, J=7.8Hz, 1H), 7.75 (s, 1H), 7.73 (d, J=8.1Hz, 1H), 7.71 (d, J=2.4Hz, 1H), 7.62 (t,
J=7.5Hz, 1H), 7.31 (dd, J=1.8, 7.8Hz, 1H), 7.23 (d, J=1.8Hz, 1H), 7.08 (d, J=8.4Hz, 1H), 3.79 (s, 3H), 3.76
(s, 3H).
13C NMR (300 MHz, DMSO-d6): δ 154.11, 153.43, 151.31, 149.17, 144.81, 143.01, 141.53, 134.10,
133.63, 130.42, 129.61, 128.54 (2C), 127.88, 122.95, 119.43, 118.31, 116.76, 115.21, 115.10, 57.21 (2C)
HRMS (m/z): [M]+ calcd for C22H18ClN3O5S, 471.0721; found, 471.5124
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3-chloro-N-(5-chloro-6-(quinolin-3-yloxy)pyridin-3-yl)-4-fluorobenzenesulfonamide (8).
1H NMR (300 MHz, DMSO-d6): δ 10.67 (s, 1H), 8.79 (d, J=2.4Hz, 1H), 8.17 (d, J=2.4Hz, 1H), 8.05 (d,
J=8.1Hz, 1H), 7.97 (d, J=2.1Hz, 1H), 7.95 (s, 1H), 7.94 (d, J=7.5Hz, 1H), 7.79 (d, J=2.4Hz, 1H), 7.77-7.73
(m, 2H), 7.72 (d, J=2.1Hz, 1H), 7.60 (d, J=9.0Hz, 1H).
13C NMR (300 MHz, DMSO-d6): δ 163.71, 153.64, 149.97, 144,83, 143.03, 141.57, 137.65, 133.67,
130.73, 129.41, 129.30, 129.08, 128.57 (2C), 127.91, 122.32, 122.11, 118.56, 118.27, 115.12
HRMS (m/z): [M]+ calcd for C20H12Cl2FN3O3S, 463.0532; found, 463.6125
N-(5-chloro-6-(quinolin-3-yloxy)pyridin-3-yl)-2,4-dimethylbenzenesulfonamide (9)
1H NMR (300 MHz, DMSO-d6): δ 10.67 (s, 1H), 8.76 (d, J=2.7Hz, 1H), 8.13 (d, J=2.7Hz, 1H), 8.04 (d,
J=8.1Hz, 1H), 7.94 (d, J=8.1Hz, 1H), 7.75-7.59 (m, 5H), 7.23 (d, J=7.5Hz, 1H), 7.18 (d, J=8.1Hz, 1H),
2.25 (s, 3H), 2.29 (s, 3H).
13C NMR (300 MHz, DMSO-d6): δ 153.61, 149.11, 144.87, 143.03, 142.57, 141.62, 137.45, 136.91,
134.46, 133.64, 130.52, 129.11, 128.53 (2C), 128.29, 127.89, 127.77, 123.04, 118.52, 115.13, 23.42, 22.61
HRMS (m/z): [M]+ calcd for C22H18ClN3O3S, 439.0821; found, 439.2824
N-(5-chloro-6-(quinolin-3-yloxy)pyridin-3-yl)-3-methylbenzenesulfonamide (10).
1H NMR (300 MHz, DMSO-d6): δ 10.53 (s, 1H), 8.77 (d, J=2.4Hz, 1H), 8.14 (d, J=2.4Hz, 1H), 8.04 (d,
J=8.1Hz, 1H), 7.94 (d, J=8.1Hz, 1H), 7.76-7.66 (m, 3H), 7.64 (d, J=7.5Hz, 1H), 7.55 (m, 1H), 7.52 (d,
J=5.4Hz, 1H), 7.44 (m, 1H), 2.25 (s, 3H).
13C NMR (300 MHz, DMSO-d6): δ 153.62, 149.13, 144.87, 142.65, 141.63, 140.53, 138.97, 133.65,
133.21, 130.54, 129.91, 129.80, 128.56 (2C), 128.41, 127.91, 127.79, 125.31, 123.03, 118.51, 115.11,
22.31
HRMS (m/z): [M]+ calcd for C21H16ClN3O3S, 425.0611; found, 425.3214
N-(5-chloro-6-(quinolin-3-yloxy)pyridin-3-yl)-2-methylbenzenesulfonamide (11).
1H NMR (300 MHz, DMSO-d6): δ 10.74 (s, 1H), 8.76 (d, J=2.7Hz, 1H), 8.14 (d, J=2.7Hz, 1H), 8.04 (d,
J=8.4Hz, 1H), 7.94 (d, J=7.8Hz, 1H), 7.75 (m, 1H), 7.73 (d, J=2.4Hz, 1H), 7.70 (d, J=2.4Hz, 1H), 7.61 (t,
J=7.8Hz, 1H), 7.53 (t, J=7.8Hz, 1H), 7.39 (t, J=7.5Hz, 1H), 7.36 (d, J=5.4Hz, 1H), 2.55 (s, 3H).
13C NMR (300 MHz, DMSO-d6): δ 153.61, 149.12, 144.87, 142.59, 141.81, 141.01, 137.43, 133.61, 12.80,
132.63, 130.71, 130.71, 130.54, 129.11, 128.52, 128.39, 127.88, 123.03, 121.79, 118.52, 115.12, 23.11
HRMS (m/z): [M]+ calcd for C21H16ClN3O3S, 425.0611; found, 425.3214
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2-chloro-N-(5-chloro-6-(quinolin-3-yloxy)pyridin-3-yl)-4-fluorobenzenesulfonamide. (12)
1H NMR (300 MHz, DMSO-d6): δ 11.01 (s, 1H), 8.77 (d, J=2.7Hz, 1H), 8.15 (d, J=2.4Hz, 1H), 8.12 (dd,
J=6.0, 9.0Hz, 1H), 8.04 (d, J=8.1Hz, 1H), 7.94 (d, J=8.1Hz, 1H), 7.76-7.71 (m, 4H), 7.62 (t, J=7.8Hz, 1H),
7.34 (td, J=8.7, 2.4Hz, 1H).
13C NMR (300 MHz, DMSO-d6): δ 169.53, 153.62, 149.12, 144.85, 143.01, 141.63, 136.32, 134.11,
133.61, 131.43, 130.52, 129.61, 128.52 (2C), 127.63, 123.02, 119.12, 118.53, 115.91, 115.21
HRMS (m/z): [M]+ calcd for C20H12Cl2FN3O3S, 463.0031; found, 463.0834
N-(5-chloro-6-(quinolin-3-yloxy)pyridin-3-yl)-2,4-difluorobenzenesulfonamide (13)
1H NMR (300 MHz, DMSO-d6): δ 10.99 (s, 1H), 8.78 (d, J=2.4Hz, 1H), 8.17 (d, J=2.4Hz, 1H), 8.04 (d,
J=8.1Hz, 1H), 7.94 (d, J=8.1Hz, 1H), 7.89-7.86 (m, 2H), 7.78-7.70 (m, 2H), 7.62 (t, J=7.2Hz, 1H), 7.52
(m, 2H), 7.29 (td, J=8.4, 2.1Hz, 1H).
13C NMR (300 MHz, DMSO-d6): δ 165.34, 160.86, 153.61, 149.19, 144.86, 143.03, 141.65, 133.27,
131.53, 130.61, 129.62, 128.54 (2C), 127.68, 123.08 (2C), 118.59, 115.96, 112.56, 106.14
HRMS (m/z): [M]+ calcd for C20H12ClF2N3O3S, 447.0612; found, 447.1304
2-chloro-N-(5-chloro-6-(quinolin-3-yloxy)pyridin-3-yl)-4-(trifluoromethyl)benzene-sulfonamide (14)
1H NMR (300 MHz, DMSO-d6): δ 10.66 (s, 1H), 8.79 (d, J=2.7Hz, 1H), 8.15 (d, J=2.7Hz, 1H), 8.05 (d,
J=8.1Hz, 1H), 7.97 (d, J=8.1Hz, 1H), 7.78-7.57 (m, 5H), 7.25 (d, J=7.5Hz, 1H), 7.19 (d, J=8.1Hz, 1H).
13C NMR (300 MHz, DMSO-d6): δ 153.62, 149.13, 144.87, 144.65, 143.03, 141.67, 141.35, 133.29,
132.83, 130.54, 130.11, 129.62, 128.55 (2C), 127.64, 126.20 (2C), 124.61, 123.10, 118.58, 115.93
HRMS (m/z): [M]+ calcd for C21H12Cl2F3N3O3S, 512.9912; found, 513.1206
N-(5-chloro-6-(quinolin-3-yloxy)pyridin-3-yl)-4-fluoro-2-methylbenzenesulfonamide (15).
1H NMR (300 MHz, DMSO-d6): δ 10.76 (s, 1H), 8.76 (d, J=2.7Hz, 1H), 8.14 (d, J=2.4Hz, 1H), 8.04 (d,
J=8.1Hz, 1H), 7.94-7.87 (m, 2H), 7.79-7.70 (m, 3H), 7.61 (td, J=7.8, 1.5Hz, 1H), 7.33 (dd, J=2.4, 9.6Hz,
1H), 7.22 (td, J=2.7, 8.4Hz, 1H), 2.58 (s, 3H).
13C NMR (300 MHz, DMSO-d6): δ 167.15, 153.61, 149.20, 144.82, 143.02, 141.66, 139.27, 135.54,
133.28, 130.53, 129.80, 129.60, 128.54 (2C), 127.63, 123.01, 118.57, 117.38, 115.92, 115.18, 23.03
HRMS (m/z): [M]+ calcd for C21H15ClFN3O3S, 443.0531; found, 443.0732
N-(5-chloro-6-(quinolin-3-yloxy)pyridin-3-yl)-3,4-difluorobenzenesulfonamide (16).
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1H NMR (300 MHz, DMSO-d6): δ 10.71 (s, 1H), 8.81 (d, J=2.4Hz, 1H), 8.19 (d, J=2.4Hz, 1H), 8.06 (d,
J=8.4Hz, 1H), 7.97 (d, J=8.1Hz, 1H), 7.87 (t, J=7.8Hz, 1H), 7.81-7.74 (m, 3H), 7.69-7.61 (m, 3H).
13C NMR (300 MHz, DMSO-d6): δ 155.78, 147.11, 146.81, 145.42, 138.74, 136.31, 134.87, 131.02,
129.46, 129.10, 128.22 (2C), 127.81, 125.82, 125.28, 119.58, 119.33, 118.23, 117.33, 117.07
HRMS (m/z): [M]+ calcd for C20H12ClF2N3O3S, 447.0331; found, 447.0735
N-(5-chloro-6-(quinolin-3-yloxy)pyridin-3-yl)-2,5-dimethoxybenzenesulfonamide (17).
1H NMR (300 MHz, DMSO-d6): δ 10.35 (s, 1H), 8.76 (d, J=2.7Hz, 1H), 8.14 (d, J=2.4Hz, 1H), 8.03 (d,
J=8.4Hz, 1H), 7.94 (d, J=7.5Hz, 1H), 7.76-7.70 (m, 3H), 7.64 (td, J=7.5, 1.2Hz, 1H), 7.24 (m, 1H), 7.19-
7.15 (m, 2H), 3.78 (s, 3H), 3.70 (s, 3H).
13C NMR (300 MHz, DMSO-d6): δ 152.51, 146.91, 146.79, 145.56, 136.13, 134.17, 131.09, 129.65,
129.31, 128.24 (2C), 127.72, 125.62, 125.44, 121.95, 119.58, 119.33, 118.34, 117.23, 117.11, 57.21, 57.08
HRMS (m/z): [M]+ calcd for C22H18ClN3O5S, 471.0712; found, 471.1316
N-(5-chloro-6-(quinolin-3-yloxy)pyridin-3-yl)-3-fluorobenzenesulfonamide (18).
1H NMR (300 MHz, DMSO-d6): δ 10.69 (s, 1H), 8.78 (d, J=2.7Hz, 1H), 8.17 (d, J=2.4Hz, 1H), 8.04 (d,
J=8.4Hz, 1H), 7.95 (d, J=8.1Hz, 1H), 7.76-7.71 (m, 3H), 7.68-7.50 (m, 5H).
13C NMR (300 MHz, DMSO-d6): δ 162.17, 153.66, 149.19, 145.81, 142.34, 142.21, 133.63, 132.57,
130.64, 129.87, 128.23 (2C), 127.41, 127.03, 123.81, 123.04, 118.76, 118.28, 115.43, 115.18
HRMS (m/z): [M]+ calcd for C20H13ClFN3O3S, 429.0421; found, 429.3152
N-(5-chloro-6-(quinolin-3-yloxy)pyridin-3-yl)-3-(trifluoromethyl)benzenesulfonamide (19)
1H NMR (300 MHz, DMSO-d6): δ 10.71 (s, 1H), 8.77 (d, J=2.4Hz, 1H), 8.16 (d, J=2.4Hz, 1H), 8.07 (d,
J=8.4Hz, 1H), 8.01 (m, 3H), 7.95 (d, J=8.1Hz, 1H), 7.83 (t, J=8.4Hz, 1H), 7.77-7.67 (m, 3H) 7.62 (t,
J=7.5Hz, 1H).
13C NMR (300 MHz, DMSO-d6): δ 152.63, 149.11, 143.86, 141.28, 141.03, 133.34, 132.27, 131.12,
130.53, 130.21, 129.62, 129.40, 128.24 (2C), 127.39, 125.15, 123.67, 122.86, 118.97, 115.54
HRMS (m/z): [M]+ calcd for C21H13ClF3N3O3S, 479.0311; found, 479.7215
3,4-dichloro-N-(5-chloro-6-(quinolin-3-yloxy)pyridin-3-yl)benzenesulfonamide (20).
1H NMR (300 MHz, DMSO-d6): δ 10.81 (s, 1H), 8.75 (d, J=2.4Hz, 1H), 8.17 (d, J=2.4Hz, 1H), 8.04 (d,
J=7.5Hz, 1H), 7.95-7.93 (m, 2H), 7.86 (d, J=7.5Hz, 1H), 7.80 (d, J=2.1Hz, 1H), 7.74 (d, J=7.5Hz, 1H),
7.72 (d, J=2.1Hz, 1H), 7.68-7.66 (dd, J=8.4, 2.1Hz, 1H), 7.62 (t, J=7.5Hz, 1H).
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13C NMR (300 MHz, DMSO-d6): δ 153.61, 149.19, 144.84, 142.97, 141.63, 140.22, 137.76, 134.73,
133.46, 131.51, 130.71, 129.33, 129.10, 128.56 (2C), 127.89, 127.78, 122.10, 117.42, 115.21
HRMS (m/z): [M]+ calcd for C22H20Cl3N3O3S, 480.9632; found, 480.9113
References:
[1] P. D. G. Miles, Y. Barak, R. M. Evans, J. M. Olefsky, Am J Physiol Endocrinol Metab 284
(2003) E618-E626
[2] G. Jiang, Q. Dallas-Yang, Z. Li, D. Szalkowski, F. Liu, X. Shen, M. Wu, G. Zhou, T. Doebber, J.
Berger, D. E. Moller, B. B. Zhang, Diabetes 51(2002) 2412–2419
[3] E. D. Rosen, P. Sarraf, A. E. Troy, G. Bradwin, K Moore, D. S. Milstone, B. M. Spiegelman, R. M.
Mortensen, Mol. Cell. 4 (1999) 611-617
[4] C. Day, Diabet. Med. 16 (1999) 179-92
[5] E. E. Kershaw, M. Schupp, H-P. Guan, N. P. Gardner, M. A. Lazar, J. S. Flier, American Journal of
Physiology - Endocrinology and Metabolism 293 (2007) E1736-E1745.
[6] Y. Barak and S. Kim, PPAR Research (2007) 1-12
[7] D. Patsouris, S. Mandard, P. J. Voshol, P. Escher, N. S. Tan, L. M. Havekes, W. Koenig, W. März, S.
Tafuri, W. Wahli, M. Müller, S. Kersten, J. Clin. Invest. 114 (2004) 94–103.
[8] D. L. Gerhold, F. Liu, G. Jiang, Z. Li, J. Xu, M. Lu, J.R. Sachs, A. Bagchi, A. Fridman, D.J.
Holder, T. W. Doebber, J. Berger, A. Elbrecht, D. E. Moller, B. B. Zhang, Endocrinology 143 (2002)
2106-2118.
[9] K. G. Tolman, J. Chandramouli, Clin Liver Dis. 7 (2003) 369-379.
[10] A.Motani, Z. Wang, J. Weiszmann, L. R. McGee, G. Lee, Q. Liu, J. Staunton, Z. Fang, H.
Fuentes, M. Lindstrom, J. Liu, D. H. Biermann, J. Jaen, N. P. Walker, R. M. Learned, J. L. Chen, Y.
Li, J Mol Biol 386 (2009) 1301-1311.
[11] V. Koch, S. Schnatterer, Synthesis 6 (1990) 499 – 501
[12] A. Motani, Z. Wang, J. Weiszmann, L. R. McGee, G. Lee, Q. Liu, J. Staunton, Z. Fang, H.
Fuentes, M. Lindstrom, J. Liu, D. H. T. Biermann, J. Jaen, N. P. C. Walker, R. M. Learned, J-L. Chen
and Y. Li, J. Mol. Biol. 386 (2009) 1301–1311
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Table 1:
Compound Ar=Substituted phenyl
Adipogenesis Activity
PPAR-γ Transactivation
Emax(% ) a
Emax (%) b
7
8
9
3,4-dimethoxybenzene
3-chloro-4-fluorobenzene
2,4-dimethylbenzene
3-methylbenzene
2-methylbenzene
2-chloro-4-fluorobenzene
2,4-difluorolbenzene
2-chloro-4-(trifluoromethyl)benzene
2-methyl-4-fluorobenzene
3,4-difluorobenzene
2,5-dimethoxybenzene
3-fluorobenzene
60
56
67
90
95
51
56
57
73
95
54
80
75
93
28
31
36
43
39
32
86
83
86
71
62
50
10
11
12
13
14
15
16
17
18
19
3-(trifluoromethyl)benzene
20
3,4-dichlorobenzene
35
56
a The compounds were tested for adipogenic activity in the presence of insulin in 3T3-L1 cells at
a concentration of 20µg/ml.
b The compounds were tested for agonist activity on PPAR-γ in transiently transfected HEK 293
cells at a concentration of 10µM.

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Table 2
Parameters
Compound 7
0.25
Compound 16
0.50
Tmax (h)
Cmax (µg/mL)
33.90
146.44
0.23
8.33
AUC0-8 (h*µg/mL)
Elimination rate constant(1/h)
Half life (h)
25.44
0.39
2.99
1.76
Volume of distribution (l/kg)
Clearance (L/h/kg)
1.23
4.78
0.28
1.87

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Caption page
Fig-1: Structure of known PPARγ agonist
Scheme-1 Synthesis of N-(5-chloro-6-(quinolin-3-yloxy)pyridin-3-yl)benzenesulfonamide
derivatives:
7, Ar = 3,4-dimethoxybenzene
8, Ar = 3-chloro-4-fluorobenzene
9, Ar = 2,4-dimethylbenzene
10, Ar = 3-methylbenzene
14, Ar = 2-chloro-4-(trifluoromethyl)benzene
15, Ar = 2-methyl-4-fluorobenzene
16, Ar = 3,4-difluorobenzene
17, Ar = 2,5-dimethoxybenzene
18, Ar = 3-fluorobenzene
11, Ar = 2-methylbenzene
12, Ar = 2-chloro-4-fluorobenzene
13, Ar = 2,4-difluorobenzene
19, Ar = 3-(trifluoromethyl)benzene
20, Ar = 3,4-dichlorobenzene
Reagents:
a. HCl / NaClO3 (aqueous), 50 oC b. POCl3 / Quinoline, 120 oC, 2 hrs
c. Cs2CO3, DMF, heating at 70-80 oC, 2-4 hrs d. SnCl2.H2O, Ethylacetate, r.t, 15 hrs
e. Pyridine, DCM, r.t, 10-15 hrs
Figure-2: Superimposition of INT131 (Brown) and compound 7 (Blue) inside the PPARγ active
pocket.
Figure-3: View from the back-side of H3 helix
Table 1: Compounds with their biological activities
Table 2: Pharmacokinetic profile of compound 7 and 16
Fig-4: Effect of compound 7 on plasma glucose in db/db mice
Fig-5: Effect of compound 7 on body weight in db/db mice

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O
N
N
S
HO
O
NH
O
O
S
NH
O
O
O
O
Rosiglitazone
Troglitazone
H
Cl
O
S
N
N
O
N
O
NH
Cl
S
Pioglitazone
O
INT-131 (Non TZD PPAR modulator)
O
Fig-1

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Figure-2

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Figure-3

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Fig 4
600
500
400
300
200
100
0
***
***
Day 0
Day 10
Control Rosiglitazone (5mg/kg b.i.d) Compound 7 (50mg/kg b.i.d)

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Fig 5
46
44
42
40
38
36
34
***
***
1
2
3
4
5
6
7
8
9
10
11
Days
Rosiglitazone (5mg/ kg b.i.d)
C ompo und 7 (50mg/ kg b.i.d)
vehicle

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Scheme-1