Synthesis and biological evaluation of novel 2,4-disubstituted quinazoline analogues as GPR119 agonists

Article abstract of DOI:10.1016/j.bmc.2012.12.013

GPR119 agonist has emerged as a promising target for the treatment of type 2 diabetes. A series of novel 2,4-disubstituted quinazoline analogues was prepared and evaluated their agonistic activity against human GPR119. The analogues bearing azabicyclic amine substituents (12a, 12c and 12g) exhibited better EC₅₀ values than that of OEA though they appeared to be partial agonists.

Full text of DOI:10.1016/j.bmc.2012.12.013

Accepted Manuscript
Synthesis and biological evaluation of novel 2,4-disubstituted quinazoline an‐
alogues as GPR119 agonists
Tuan-Anh N. Pham, Zunhua Yang, Yuanying Fang, Jun Luo, Jongkook Lee,
Haeil Park
PII:
S0968-0896(12)00974-1
DOI:
http://dx.doi.org/10.1016/j.bmc.2012.12.013
BMC 10479
Reference:
To appear in:
Bioorganic & Medicinal Chemistry
Received Date:
Revised Date:
Accepted Date:
17 October 2012
2 December 2012
3 December 2012
Please cite this article as: Pham, T.N., Yang, Z., Fang, Y., Luo, J., Lee, J., Park, H., Synthesis and biological
evaluation of novel 2,4-disubstituted quinazoline analogues as GPR119 agonists, Bioorganic & Medicinal
Chemistry (2012), doi: http://dx.doi.org/10.1016/j.bmc.2012.12.013
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Synthesis and biological evaluation of novel 2,4-disubstituted
quinazoline analogues as GPR119 agonists
Tuan-Anh N. Phama, Zunhua Yanga,b, Yuanying Fanga, Jun Luoa, Jongkook Leea, Haeil Parka,*
aCollege of Pharmacy, Kangwon National University, Chunchon 200-701, Korea
bCollege of Pharmacy, Jiangxi University of Traditional Chinese Medicine, Nanchang 330004, China
ABSTRACT
GPR119 agonist has emerged as a promising target for the treatment of type 2 diabetes. A series of novel
2,4-disubstituted quinazoline analogues was prepared and evaluated their agonistic activity against human
GPR119. The analogues bearing azabicyclic amine substituents (12a, 12c and 12g) exhibited better EC50
values than that of OEA though they appeared to be partial agonists.
Keywords: GPR119 agonist, type 2 diabetes, quinazoline analogues, azabicyclic amines
1. Introduction
Type 2 diabetes, formerly known as non-insulin-dependent diabetes mellitus, is a complex metabolic
disorder that affects millions of people worldwide. It is characterized by high blood glucose that caused by
insulin resistance or insufficient insulin secretion, accounting for approximately 90% of all cases of
diabetes.1 Currently, metformin is generally recommended as a first line treatment for type 2 diabetes
beside others such as sulfonyl ureas, thiazolidinediones, α-glucosidase inhibitors,1 and the two most
recently approved classes: glucagon-like-pepetide 1 (GLP-1) receptor agonists,1-3 and dipeptidyl peptidase-
4 (DPP-4) inhibitors.2,3 Although the number of patients with type 2 diabetes that successfully achieve
target glucose levels is steadily improving, a substantial number of subjects continue to fall short of
acceptable treatment goals, leaving them at high risk of development of diabetes-associated complications.
Furthermore, most of these medications remain side effects such as hypoglycemia, weight gain, bone
fractures.1,2 Therefore, new drugs that exhibit improved efficacy and safety relative to current available
medications are clearly needed.
GPR119 is a G-protein-coupled receptor predominantly express on pancreatic beta cells and intestinal
enteroendocrine cells. GPR119 can be activated by oleoylethanolamine (OEA), lysophosphatidylcholine,
N-oleoyldopamin, ovalnil…and OEA most probably represents the endogenous ligand.4-7 GPR119 agonist
stimulates glucose-dependent insulin secretion in vitro and lowers an elevated blood glucose level in
vivo.5,6 Furthermore, they have been demonstrated to stimulate the release of the incretin (GLP-1 and
GIP).5-7 Therefore, GPR119 agonist has emerged as a promising target for the treatment of type 2 diabetes
and obesity by improving glucose homoeostasis while concurrently slowing gastric emptying, reducing
food intake and promoting weight loss.4-6 To date, several candidates were entered clinical trials for
treatment of type 2 diabetes. However, very limited information is yet available from these trials in the
form of scientific publications.6,7
Diverse fused aromatic heterocycle scaffolds were found in the precedent literatures and patents of
GPR119 agonists (Fig. 1).6-14 As a congener of bicyclic heterocycle scaffolds, we were interested in
quinazoline scaffold since quinazoline derivatives were found to possess diverse promising biological
activities including anticancer, antihypertension, anticonvulsant, antidiabetic, antimalarial etc.15-17 Some of
them are used in clinical treatment such as terazosin, alfuzosin, doxazosin,15 gefitinib, erlotinib.16 Also
piperidine moieties were attached to the aromatic heterocycle scaffolds in most compounds under
development.6-13 It was suggested from precedent literatures that piperidine carbamate region makes H-
bond acceptor interactions that plays very important role for GPR119 agonistic activity. We also found that
modifications in this region may affect to bioactivity. Therefore, we designed analogs bearing
conformationally restricted azabicyclic rings instead of piperidines to evaluate effect on GPR119 activation.
Thus we designed novel quinazoline derivatives bearing piperidine and azabicyclic analogues and herein
we report the synthesis, biological evaluation and structural-activity relationship of novel 2,4-disubstituted
quinazoline analogues as GPR119 agonists for the treatment of type 2 diabetes.
- 1 -

F
H CO S
3
2
N
N
O
O
N
N
N
O
N
O
N
SO CH
N
O
2
3
N
N
H
F
N
NO
2
N
N
N
O
N
O
O
AR231453
PSN632408
APD668
N
O
N
N
N
N
H
H CO S
O
O
3
2
N
N
F
N
N
OH
N
H
N
OH
Cl
F
GSK1292263
Fig. 1. Structures of selected synthetic GPR119 agonists
2. Results and discussion
2.1. Chemistry
The synthesis of 2-arylamino-4-alkyloxy-6,7-dimethoxyquinazoline 3a-d are outlined in Scheme 1. N-Boc-
4-hydroxypiperidine, N-Boc-4-hydroxymethylpiperidine and 4-methylsulfonylaniline are commercial
available products. 2-Fluoro-4-methylsulfonylaniline was prepared according to previously reported
procedure.18 Incorporating N-Boc piperidine alcohols into 2,4-dichloro-6,7-dimethoxyquinazoline in the
presence of NaH yielded 4-alkoxy-2-chloro-6,7-dimethoxyquinazoline (1 and 2).11 Coupling reactions of
these intermediates (1 and 2) with 4-methylsulfonylaniline and 2-fluoro-4-methylsulfonylaniline under acid
catalyst19 followed by treatment with Boc2O in the presence of TEA gave target products 3a-d.20
SO CH
2
3
O
O
N
O
Cl
X
NH
2
N
OH
n
O
O
N
O
NH
X
O
O
N
Cl
b or c
a
N
+
+
N
n
N
Boc
SO CH
2
3a (n = 0, X = H)
3b (n = 1, X = H)
3c (n = 0, X = F)
3d (n = 1, X = F)
Cl
3
n
(n=0,1)
N
1 (n = 0)
Boc 2 (n = 1)
(X=H,F)
N
Boc
Scheme 1. Synthesis of 2-arylamino-4-alkyloxy-6,7-dimethoxyquinazolines (3a-d). Reagents and
conditions: (a) NaH, THF, rt, overnight (b) (i) HCl, n-BuOH, reflux, overnight (ii) Boc2O, TEA, CH2Cl2, rt,
2h (for 3a, 3b) (c) (i) HCl, AcOH, reflux, overnight (ii) Boc2O, TEA, CH2Cl2, rt, 2h (for 3c, 3d).
Quinazoline analogues 6a-d were synthesized according to Scheme 2. 2,4-Dichloro-6,7-
dimethoxyquinazoline was reacted with 4-methylsulfonylaniline under acid catalyst to afford a key
intermediate 4.21 By contrast, another key intermediate 5 was achieved with 2,4-dichloro-6,7-
dimethoxyquinazoline and 2-fluoro-4-methylsulfonylanilne in the presence of NaH. Finally, these
intermediates were converted into desire products 6a-d by treatment with N-Boc piperidine alcohols using
NaOH as a base.
Boc
N
n
O
O
O
N
Cl
NH
2
OH
n
O
O
N
O
O
N
Cl
X
N
a or b
c
+
+
N
N
NH
N
Boc
NH
X
Cl
SO CH
2
3
6a (n = 0, X = H)
6b (n = 1, X = H)
6c (n = 0, X = F)
6d (n = 1, X = F)
H CO S
X
3
2
H CO S
(X=H,F)
3
2
4 (X = H)
5 (X = F)
(n=0,1)
- 2 -

Scheme 2. Synthesis of 2-alkyloxy-4-arylamino-6,7-dimethoxyquinazolines (6a-d). Reagents and
o
conditions: (a) HCl, EtOH, 60 C, overnight (for 6a, 6b) (b) NaH, THF, reflux, overnight (for 6c, 6d) (c)
NaOH, 140 oC, overnight.
Azabicyclic amines (7a-c) were generated following the previously reported procedures and 1H NMR data
were consistent with that reported in the literature.22 4-Alkylamino-2-chloro-6,7-dimethoxyquinazolines (8-
10), obtained from 2,4-dichloro-6,7-dimethoxyquinazoline and azabicyclic amines (7a-c)23 were reacted
with 4-methylsulfonylaniline to afford product 11a-c19 while the reactions between 4-Alkylamino-2-
chloro-6,7-dimethoxyquinazolines (8-10) and 2-fluoro-4-methylsulfonylanilne did not yield target products.
Hydrogenolysis of compound 11c followed by reaction with Boc2O yielded compound 11d (Scheme 3).
SO CH
2
3
NH
2
O
O
N
R
Cl
O
O
N
R
NH
O
O
N
Cl
a
b
N
+
N
N
7a, 7b, 7c
SO CH
Cl
2
3
11a (R = 7a)
11b (R = 7b)
11c (R = 7c)
11d (R = 7d)
8
9
(R = 7a)
(R = 7b)
10 (R = 7c)
c
X
N
NH
2
H N
2
N
N
Boc
HN
R-H =
Boc
7a
7b
7c (X = Bn)
7d (X = Boc)
Scheme 3. Synthesis of 4-alkylamino-2-(4-methylsulfonylphenylamino)-6,7-dimethoxyquinazolines (11a-
d). Reagents and conditions: (a) Hunig’s base, i-PrOH, rt, overnight (b) (i) HCl, n-BuOH, reflux, overnight
(ii) Boc2O, TEA, CH2Cl2, rt, 2h (for 11a, 11b) (c) (i) H2, Pd/C, MeOH, rt, 5h (ii) Boc2O, TEA, CH2Cl2, rt,
2h.
2-Alkylamino-4-arylamino-6,7-dimethoxyquinazolines 12a-e were synthesized from intermediates (4 and
5) and azabicyclic amines (7a-c) in the presence of Hunig’s base as a catalyst.21 Hydrogenolysis of
compounds (12d and 12e) followed by treatment with Boc2O yielded compounds (12f and 12g) (Scheme 4).
O
O
N
R
O
O
N
Cl
a
N
N
7a, 7b, 7c
NH
X
NH
X
H CO S
H CO S
3
2
3
2
4
5
(X = H)
(X = F)
12a (R = 7a, X = F)
12b (R = 7b, X = H)
12c (R = 7b, X = F)
12d (R = 7c, X = H); 12e (R = 7c, X = F)
12f (R = 7d, X = H); 12g (R = 7d, X = F)
b
Scheme 4. Synthesis of 2-alkylamino-4-arylamino-6,7-dimethoxyquinazolines (12a-g). Reagents and
conditions: (a) Hunig’s base, n-BuOH, reflux, 1 day (b) (i) H2, Pd/C, MeOH, rt, 5h (ii) Boc2O, TEA,
CH2Cl2, rt, 2h.
2.2. Biological activity
Compounds 3a-d, 6a-d, 11a-d and 12a-g were evaluated their abilities to active the human GPR119
receptor in a cell-based cAMP assay and expressed in EC50 and %max values. The EC50 values represent
- 3 -

the concentration of the tested compounds for 50% cAMP of OEA, while the %max values present the
relative response (%) of the tested compounds compared to the maximal effect of OEA.
As shown in Table 1, the quinazoline analogues (3a, 6a), possessing N-Boc-4-piperidinol and 4-
methylsulfonylphenylamino groups either at C-2 or C-4 positions, were identified as weak GPR119
agonists (EC50> 10 µM and %max=20.6, 17.9, respectively) and their agonistic activities were much less
potent than OEA. Replacement of N-Boc-4-piperidinol with N-Boc-4-piperidinemethanol also gave similar
results. The compounds 3b and 6b exhibited weak to negligible GPR119 activation activities. The
derivatives with (2-fluoro-4-methylsulfonyl)phenylamino group (3c, 3d, 6c, 6d) showed similar GPR119
activation activities. These results indicated that the position of (2-fluoro-4-methylsulfonyl)phenylamino
and heterocyclic alcohol groups did not significantly affect to GPR119 activation activity.
Table 1
In vitro GPR119 agonistic activities of 3a-d and 6a-d
SO CH
2
3
X
Boc
N
O
O
N
O
NH
N
n
O
O
N
O
n
N
NH
N
Boc
H CO S
X
3
2
3a-d
6a-d
Compound
X
n
hGPR119 activity
EC50a (µM) %maxb
>10 20.6
H
H
H
H
F
F
F
F
0
0
1
1
0
0
1
1
3a
6a
3b
6b
3c
6c
3d
6d
OEA
>10
>10
NEc
>10
>10
>10
>10
2.2
17.9
17.4
NEc
19.7
19.6
21.3
33.2
100
a concentration for 50% cAMP stimulation of OEA
b cAMP stimulation % compared to maximal effect of OEA
c Not effective at 10 µM
To explore the steric influence on bioactivity, analogues bearing azabicyclic amines such as 7a-d instead of
4-piperidinol and piperidine-4-methanol were prepared and evaluated their hGPR119 agonistic activities as
shown in table 2. The quinazoline analogues (11a-d, 12b, 12d and 12f), possessing azabicyclic amines and
4-methylsulfonylphenylamino groups either at C-2 or C-4 positions, did not improve agonistic activity
compared to those of analogues with heterocyclic alcohols. However, the analogues (12a, 12c and 12g)
bearing both (2-fluoro-4-methylsulfonyl)phenylamino and azabicyclic amine groups showed strong
agonistic activities and exhibited better EC50 values than that of OEA but they were proved as partial
agonists as shown by their relatively low %max values. These results suggest that the combination of
fluorine atom on benzene ring and azabicyclic amines proved to have significantly synergistic effect. These
analogues (12a, 12c and 12g) possessed equi-potential for agonistic activities as shown by similar values of
EC50 and %max. From these findings, we speculated that while the sterically bulky of azabicyclic
carbamate would likely be favorable for agonistic activity, the length of this region (one or two carbon
elongation) as well as the configuration (endo, exo) were not essential for the in vitro hGPR119 agonistic
- 4 -

activity. Also it was observed that the analogue with N-Boc exhibited better agonistic activity than that of
the analogue with N-Bn (12g vs. 12e) suggesting that hydrogen-bond acceptor interactions in this region
was important for showing potent agonistic activity.
Table 2
In vitro GPR119 agonistic activities of 11a-d and 12a-g
SO CH
2
3
O
O
N
R
N
O
O
N
R
NH
NH
X
N
H CO S
3
2
11a-d
12a-g
Compound
X
R
hGPR119 activity
EC50a (µM)
%maxb
17.0
-
>10
11a
12a
NH
F
1.0
46.2
N
Boc
-
>10
>10
1.0
15.1
19.9
53.7
11b
12b
12c
H
N
H
F
N
Boc
Bn
N
-
>10
>10
>10
32.8
23.4
44.3
11c
12d
12e
H
F
N
Boc
N
-
>10
>10
1.0
37.5
32.7
56.9
100
11d
12f
H
F
N
12g
2.2
OEA
a concentration for 50% cAMP stimulation of OEA
b cAMP stimulation % compared to maximal effect of OEA
3. Conclusion
In summary, we have synthesized and characterized a series of novel 2,4-disubstituted quinazolines with
heterocyclic alcohols and azabicyclic amine groups as GPR119 agonists for the treatment of type 2 diabetes.
We found that the analogues bearing (2-fluoro-4-methylsulfonyl)phenylamino and azabicyclic amine
groups with N-Boc (12a, 12c and 12g) exhibited better EC50 values than that of OEA even though they
appeared to be partial agonists. Our present results suggest that the steric bulkyness of introduced groups
seems to influence on bioactivity (3c, 3d, 6c, 6d vs. 12a, 12c, 12g) while the configuration (endo vs. exo)
affects not much on bioactivity (12a vs. 12c). The electronic effects of the substituted groups at 2-position
of benzene ring (11a vs. 12a; 12b vs. 12c; 12f vs. 12g) as well as N-Boc group of azabicyclic amines (12e
- 5 -

vs. 12g) may play a very important role for better bioactivity. Further SAR research to discover more potent
GPR119 agonists with diverse heterocyclic scaffolds is under progress.
4. Experimental section
1
All starting materials were obtained from commercial suppliers and used without further purification. H
NMR spectra were recorded on a Bruker DPX 300 (300 MHz) spectrometer. Chemical shifts are reported
in parts per million (ppm) downfield relative to tetramethylsilane as an internal standard. Peak splitting
patterns are abbreviated as s (singlet), brs (broad singlet), d (doublet), t (triplet), dd (doublet of doublet) and
m (multiplet). MALDI-TOF mass spectra were recorded on a Voyager-DE STR. Melting points were
recorded on a Fisher-Johns microscopic scale melting point apparatus. TLC was performed on silica F254
and detected by UV light at 254 nm or by charring with either ninhydrin or iodine. Column
chromatography was performed on Kieselgel 60 (230~400 mesh, Merck).
4.1. General procedure to generate 4-alkoxy-2-chloro-6,7-dimethoxyquinazoline (1-2)
To a stirred solution of sodium hydride (4.0 mmol) in dry THF (3 ml) was added solution of alcohols (2.4
mmol) in dry THF (6 ml) and the resulting reaction mixture was stirred for 1 hour under N2 gas. The
reaction mixture was cooled to 0 oC and a solution of 2,4-dichloro-6,7-dimethoxyquinazoline (2.0 mmol) in
dry THF (6 ml) was added and stirred for overnight. The reaction was quenched with water and extracted
with dichloromethane. The combined organic layer was dried over anhydrous MgSO4, filtered,
concentrated under reduced pressure. The crude residue was purified by silica gel chromatography (hexane-
acetone 20:1).
4.1.1. tert-Butyl 4-(2-chloro-6,7-dimethoxyquinazolin-4-yloxy)piperidine-1-carboxylate (1)
Following the general procedure, the title compound was prepared from 2,4-dichloro-6,7-
dimethoxyquinazoline and N-Boc-4-hydroxypiperidine in 64% yield; white solid; mp 190-191 oC; 1H NMR
(300MHz, CDCl3): δ 7.28 (s, 1H), 7.20 (s, 1H), 5.61-5.53 (m, 1H), 4.02 (s, 6H), 3.88-3.84 (m, 2H), 3.40-
3.32 (m, 2H), 2.16-2.07 (m, 2H), 1.93-1.82 (m, 2H), 1.47 (s, 9H).
4.1.2. tert-Butyl 4-((2-chloro-6,7-dimethoxyquinazolin-4-yloxy)methyl)piperidine-1-carboxylate (2)
Following the general procedure, the title compound was prepared from 2,4-dichloro-6,7-
o
1
dimethoxyquinazoline and N-Boc-4-hydroxmethylypiperidine in 73% yield; white solid; mp 176 C ; H
NMR (300MHz, CDCl3): δ 7.27 (s, 1H), 7.21 (s, 1H), 4.45 (d, J = 6.6 Hz, 2H), 4.21 (brs, 2H), 4.02 (s, 6H),
2.79 (t, J = 2.2 Hz, 2H), 2.14-2.10 (m, 1H), 1.86 (d, J = 2.5 Hz, 2H), 1.47 (s, 9H), 1.42-1.29 (m, 2H).
4.2. General procedure to generate 2-arylamino-4-alkyloxy-6,7-dimethoxyquinazoline (3a-d)
To the solution of 4-alkyloxy-2-dichloro-6,7-dimethoxyquinazoline (1 or 2, 1.0 mmol) in n-BuOH (for 3a
and 3b) or AcOH (for 3c and 3d) were added arylamine (1.5 mmol) and 1N HCl aqueous solution (2.0
o
mmol) and the reaction mixture was stirred at 130 C for overnight. The reaction mixture was cooled to
room temperature then neutralized with 1N NaOH aqueous solution and concentrated under reduced
pressure. To the solution of crude product in CH2Cl2 were added TEA (2.0 mmol) and Boc2O (1.5 mmol)
and the reaction mixture was stirred for 2 hours. After extraction with CH2Cl2, the combined organic layer
was dried over anhydrous MgSO4, filtered, concentrated under reduced pressure. Purification of the crude
product by silica gel chromatography (CHCl3-MeOH 200:1) yielded product.
4.2.1. tert-Butyl 4-(6,7-dimethoxy-2-(4-(methylsulfonyl)phenylamino)quinazolin-4-yloxy)piperidine-1-
carboxylate (3a)
Following the general procedure, the title compound was prepared from compound 1 and 4-
o
1
methylsulfonylaniline in 65% yield; white solid; mp 159-161 C; H NMR (300MHz, CDCl3): δ 7.99-7.96
(m, 2H), 7.91-7.88 (m, 2H), 7.37 (s, 1H), 7.26 (s, 1H), 7.08 (s, 1H), 5.50-5.45 (m, 1H), 4.03 (s, 3H), 4.00 (s,
3H), 3.90-3.85 (m, 2H), 3.42-3.33 (m, 2H), 3.06 (s, 3H), 2.15-2.08 (m, 2H), 1.96-1.86 (m, 2H), 1.5 (s, 9H);
13C NMR (100 MHz, CDCl3): δ 165.70, 156.30, 155.18, 154.85, 149.97, 148.01, 145.65, 132.54, 129.12,
118.27, 107.20, 106.01, 102.30, 80.23, 72.49, 56.65, 56.55, 45.30, 31.06, 28.85; MS (MALDI-TOF, 2,5-
dihydroxybenzoic acid as matrix): m/z (100%): 558.2002 M+.
- 6 -

4.2.2. tert-Butyl 4-((6,7-dimethoxy-2-(4-(methylsulfonyl)phenylamino)quinazolin-4-
yloxy)methyl)piperidine-1-carboxylate (3b)
Following the general procedure, the title compound was prepared from compound 2 and 4-
methylsulfonylaniline in 60% yield; white solid; mp 140-142 oC; 1H NMR (300MHz, CDCl3): δ 8.00-7.97
(m, 2H), 7.91-7.88 (m, 2H), 7.39 (s, 1H), 7.26 (s, 1H), 7.09 (s, 1H), 4.40 (d, J = 6.5 Hz, 2H), 4.22 (brs, 2H),
4.04 (s, 3H), 4.00 (s, 3H), 3.06 (s, 3H), 2.83-2.76 (m, 2H), 2.14-2.09 (m, 1H), 1.89-1.82 (m, 2H), 1.48 (s,
9H), 1.43-1.26 (m, 2H); MS (MALDI-TOF, 2,5-dihydroxybenzoic acid as matrix): m/z (100%): 572.2022
M+.
4.2.3.
tert-Butyl
4-(2-(2-fluoro-4-(methylsulfonyl)phenylamino)-6,7-dimethoxyquinazolin-4-
yloxy)piperidine-1-carboxylate (3c)
Following the general procedure, the title compound was prepared from compound 1 and 2-fluoro-4-
o
1
methylsulfonylaniline in 15% yield; yellow solid; mp 204 C; H NMR (300MHz, CDCl3): δ 9.14 (t, J =
8.2Hz, 1H), 7.79 (dd, J = 1.3 Hz, 8.7 Hz, 1H), 7.69 (dd, J = 2.0 Hz, 10.6 Hz, 1 H), 7.42 (d, J = 3.0 Hz, 1H),
7.27 (s, 1H), 7.11 (s, 1H), 5.53-5.48 (m, 1H), 4.06 (s, 3H), 4.00 (s, 3H), 3.86 (brs, 2H), 3.43-3.34 (m, 2H),
3.08 (s, 3H), 2.15-2.10 (m, 2H), 1.96-1.86 (m, 2H), 1.50 (s, 9H); MS (MALDI-TOF, 2,5-dihydroxybenzoic
acid as matrix): m/z (100%): 599.2000 [M+Na]+.
4.2.4. tert-Butyl 4-((2-(2-fluoro-4-(methylsulfonyl)phenylamino)-6,7-dimethoxyquinazolin-4-
yloxy)methyl)piperidine-1-carboxylate (3d)
Following the general procedure, the title compound was prepared from compound 2 and 2-fluoro-4-
methylsulfonylaniline in 14% yield; yellow solid; mp 120-123 oC; 1H NMR (300MHz, CDCl3): δ 9.14 (t, J
= 8.2 Hz, 1H), 7.79 (dd, J = 1.2 Hz, 8.7 Hz, 1H), 7.68 (dd, J = 2.0 Hz, 10.6 Hz, 1H), 7.40 (d, J = 3.9 Hz,
1H), 7.27 (s, 1H), 7.11 (s, 1H), 4.42 (d, J = 6.6 Hz, 2H), 4.22-4.19 (m, 2H), 4.05 (s, 3H), 4.04 (s, 3H), 3.07
(s, 3H), 2.80 (t, J = 11.5 Hz, 2H), 2.14-2.12 (m, 1H), 1.87 (d, J = 12.6 Hz, 2H), 1.48 (s, 9H), 1.46-1.31 (m,
2H); MS (MALDI-TOF, 2,5-dihydroxybenzoic acid as matrix): m/z (100%): 613.2001 [M+Na]+.
4.3. 2-Chloro-4-(4-methylsulfonylphenylamino)-6,7-dimethoxyquinazoline (4)
To the solution of 2,4-dichloro-6,7-dimethoxyquinazoline (0.52 g, 2.0 mmol) in EtOH (20 ml) were added
4-methylsulfonylaniline (0.34 g, 2.0 mmol) and 1N HCl aqueous solution (2 ml) and the reaction mixture
o
was stirred at 60 C for overnight. The reaction mixture was neutralized with 1N NaOH aqueous solution
and the precipitate was filtered and washed with CH2Cl2 to obtain product as a white solid in 75% yield; mp
o
1
240 C; H NMR (300MHz, DMSO): δ 10.17 (s, 1H), 8.3-8.1 (m, 2H), 8.05-8.02 (m, 2H), 7.94 (s, 1H),
7.27 (s, 1H), 4.03 (s, 3H), 4.00 (s, 3H), 3.30 (s, 3H).
4.4. 2-Chloro-4-(2-fluoro-4-methylsulfonylphenylamino)-6,7-dimethoxyquinazoline (5)
To a stirred solution of sodium hydride (0.05 g, 2.1 mmol) in dry THF (2 ml) was added a solution of 2-
fluoro-4-methylsulfonylaniline (0.2 g, 1.1 mmol) in dry THF (5 ml). The reaction mixture was stirred for 1
hour under N2 gas. The solution of 2,4-dichloro-6,7-dimethoxyquinazoline (0.3 g, 1.2 mmol) in dry THF (2
ml) was added to the reaction mixture at 0 oC and refluxed for overnight. The reaction was cooled to room
temperature, quenched with water and extracted with EtOAc. The combined organic layer was dried over
anhydrous MgSO4, filtered and concentrated under reduced pressure. Purification of the crude residue by
silica gel chromatography (CHCl3-MeOH 200:1) yielded product as a yellow solid in 75% yield; mp 270
oC; 1H NMR (300MHz, CDCl3+MeOD): δ 8.71 (t, J = 7.5 Hz, 1H), 7.83 (d, J = 5.7 Hz, 1H), 7.75 (dd, J =
1.3 Hz, 10.1 Hz, 1H), 7.25-7.32 (m, 2H), 4.09 (s, 3H), 4.04 (s, 3H), 3.12 (s, 3H).
4.5. General procedure to generate 2-alkyloxy-4-arylamino-6,7-dimethoxyquinazoline (6a-d)
NaOH (5 mmol) and alcohol (3.0 mmol) were added to a flask containing 2-chloro-4-arylamino-6,7-
dimethoxyquinazoline (4 or 5, 1.0 mmol) and the resulting mixture was heated at 140 oC for overnight then
cooled to room temperature. The residue was purified by silica gel chromatography (CHCl3-MeOH 200:1)
to obtain product.
4.5.1. tert-Butyl 4-(6,7-dimethoxy-4-(4-(methylsulfonyl)phenylamino)quinazolin-2-yloxy)piperidine-1-
carboxylate (6a)
Following the general procedure, the title compound was prepared from compound 4 and N-Boc-4-
1
hydroxypiperidine in 22% yield; white solid; mp 182 oC; H NMR (300MHz, CDCl3): δ 7.95 (s, 1H), 7.88-
- 7 -

7.80 (m, 4H), 7.22 (s, 1H), 7.09 (s, 1H), 5.25-5.20 (m, 1H), 4.02 (s, 6H), 3.88-3.83 (m, 2H), 3.31-3.23 (m,
2H), 3.07 (s, 3H), 2.05-2.04 (m, 2H), 1.85-1.77 (m, 2H), 1.48 (s, 9H); MS (MALDI-TOF, 2,5-
dihydroxybenzoic acid as matrix): m/z (100%): 581.2015 [M+Na]+.
4.5.2. tert-Butyl 4-((6,7-dimethoxy-4-(4-(methylsulfonyl)phenylamino)quinazolin-2-
yloxy)methyl)piperidine-1-carboxylate (6b)
Following the general procedure, the title compound was prepared from compound 4 and N-Boc-4-
hydroxymethylpiperidine in 26% yield; white solid; mp 156 oC; 1H NMR (300MHz, CDCl3): δ 8.14 (s, 1H),
7.88-7.86 (m, 2H), 7.80-7.78 (m, 2H), 7.30 (s, 1H), 7.10 (s, 1H), 4.25 (d, J = 6.6 Hz, 2H), 4.13 (brs, 2H),
4.01 (s, 3H), 4.00 (s, 3H), 3.05 (s, 3H), 2.82-2.72 (m, 2H), 2.02-1.97 (m, 1H), 1.87-1.82 (m, 2H), 1.46 (s,
9H), 1.34-1.20 (m, 2H); 13C NMR (100 MHz, CDCl3): δ 166.50, 156.23, 155.23, 154.85, 149.82, 148.00,
145.67, 132.49, 129.11, 118.30, 106.97, 106.04, 102.25, 79.91, 71.35, 56.64, 56.57, 45.30, 36.13, 29.31,
28.86; MS (MALDI-TOF, 2,5-dihydroxybenzoic acid as matrix): m/z (100%): 595.2114 [M+Na]+.
4.5.3.
tert-Butyl
4-(4-(2-fluoro-4-(methylsulfonyl)phenylamino)-6,7-dimethoxyquinazolin-2-
yloxy)piperidine-1-carboxylate (6c)
Following the general procedure, the title compound was prepared from compound 5 and N-Boc-4-
o
1
hydroxypiperidine in 29% yield; yellow solid; mp 114 C; H NMR (300MHz, CDCl3): δ 8.90 (t, J = 7.8
Hz, 1H), 7.82 (dd, J = 1.1 Hz, 8.6 Hz, 1H), 7.75 (dd, J = 2.0 Hz, 10.3 Hz, 1H), 7.62 (d, J = 4.3 Hz, 1H),
7.24 (s, 1H), 6.99 (s, 1H), 5.31-5.23 (m, 1H), 4.05 (s, 3H), 4.04 (s, 3H), 3.92-3.81 (m, 2H), 3.34-3.26 (m,
2H), 3.10 (s, 3H), 2.08 (brs, 2H), 1.88-1.84 (m, 2H), 1.49 (s, 9H); MS (MALDI-TOF, 2,5-
dihydroxybenzoic acid as matrix): m/z (100%): 599.2029 [M+Na]+.
4.5.4. tert-Butyl 4-((4-(2-fluoro-4-(methylsulfonyl)phenylamino)-6,7-dimethoxyquinazolin-2-
yloxy)methyl)piperidine-1-carboxylate (6d)
Following the general procedure, the title compound was prepared from compound 5 and N-Boc-4-
hydroxymethylpiperidine in 14% yield; yellow solid; mp 162 oC; 1H NMR (300MHz, CDCl3): δ 8.98 (t, J =
8.4 Hz, 1H), 7.84-7.81 (m, 1H), 7.76 (dd, J = 2.0 Hz, 10.4 Hz, 1H), 7.57 (d, J = 4.6 Hz, 1H), 7.14 (s, 1H),
6.96 (s, 1H), 4.30 (d, J = 6.5 Hz, 2H), 4.19 (brs, 2H), 4.04 (s, 3H), 4.03 (s, 3H), 3.09 (s, 3H), 2.78 (t, J =
12.1 Hz, 2H), 2.08-2.05 (m, 1H), 1.92-1.87 (m, 2H), 1.47 (s, 9H), 1.36-1.30 (m, 2H); MS (MALDI-TOF,
2,5-dihydroxybenzoic acid as matrix): m/z (100%): 613.1971 [M+Na]+.
4.6. General procedure to generate 4-alkylamino-2-chloro-6,7-dimethoxyquinazoline (8-10)
Azabicyclic amine (7a-c, 1.2 mmol) and Hunig’s base (2.0 mmol) were added to a solution of 2,4-dichloro-
6,7-dimethoxyquinazoline (1.0 mmol) in i-PrOH (4 ml) and the resulting mixture was stirred at room
temperature for overnight and then concentrated in vacuo. The residue was purified by column
chromatography (CHCl3-MeOH 200:1) to obtain product.
4.6.1. tert-Butyl endo-3-(2-chloro-6,7-dimethoxyquinazolin-4-ylamino)-8-azabicyclo[3.2.1]octane-8-
carboxylate (8)
Following the general procedure, the title compound was prepared from 2,4-dichloro-6,7-
o
dimethoxyquinazoline and compound 7a in 80% yield; white solid; mp 125 C; 1H NMR (300 MHz,
CDCl3): δ 7.12 (s, 1H), 6.75 (s, 1H), 5.92 (d, J = 6.4 Hz, 1H), 4.54 (m, 1H), 4.30 (brs, 2H), 3.98 (s, 6H),
2.50-1.80 (m, 8H), 1.50 (s, 9H).
4.6.2. tert-Butyl exo-3-((2-chloro-6,7-dimethoxyquinazolin-4-ylamino)methyl)-8-
azabicyclo[3.2.1]octane-8-carboxylate (9)
Following the general procedure, the title compound was prepared from 2,4-dichloro-6,7-
o
1
dimethoxyquinazoline and compound 7b in 82% yield; yellow solid; mp 242 C; H NMR (300 MHz,
CDCl3): δ 7.20-7.05 (m, 3H), 4.20 (brs, 2H), 3.90 (s, 6H), 3.10 (brs, 1H), 2.42 (m, 1H), 2.00-1.20 (m, 18H).
4.6.3. (±)-4-(9-benzyl-3,9-diazabicyclo[4.2.1]nonan-3-yl)-2-chloro-6,7-dimethoxyquinazoline (10)
Following the general procedure, the title compound was prepared from 2,4-dichloro-6,7-
o
1
dimethoxyquinazoline and compound 7c in 81% yield; yellow solid; mp 134 C; H NMR (300 MHz,
CDCl3): δ 7.40-7.25 (m, 6H), 7.16 (s, 1H), 4.42 (brs, 1H), 4.00-3.90 (m, 8H), 3.80 (s, 2H), 3.60-3.30 (m,
3H), 2.40-1.50 (m, 6H).
- 8 -

4.7. General procedure to generate 4-alkylamino-2-(4-methylsulfonylphenylamino)-6,7-
dimethoxyquinazoline (11a-c)
To the solution of 4-alkylamino-2-chloro-6,7-dimethoxyquinazoline (8-10, 1.0 mmol) in n-BuOH were
added 4-methylsulfonylaniline (1.5 mmol) and 1N HCl aqueous solution (2 mmol) and the reaction mixture
was stirred at 130 oC for overnight. The reaction mixture was cooled to room temperature, neutralized with
1N NaOH aqueous solution and concentrated under reduced pressure. To the solution of crude product in
CH2Cl2 were added TEA (2.0 mmol) and Boc2O (1.5 mmol) and the reaction mixture was stirred for 2
hours. After extraction with CH2Cl2, the combined organic layer was dried over anhydrous MgSO4, filtered,
concentrated under reduced pressure. Purification of the crude product by silica gel chromatography
(CHCl3-MeOH 200:1) yielded product.
4.7.1. tert-Butyl endo-3-(6,7-dimethoxy-2-(4-(methylsulfonyl)phenylamino)quinazolin-4-ylamino)-8-
azabicyclo[3.2.1]octane-8-carboxylate (11a)
Following the general procedure, the title compound was prepared from compound 8 and 4-
methylsulfonylaniline in 77% yield; yellow solid; mp 168-170 oC; 1H NMR (300 MHz, CDCl3): δ 7.90 (m,
4H), 7.70 (s, 1H), 7.00 (s, 1H), 6.80 (s, 1H), 5.90 (s, 1H), 4.50-4.20 (m, 3H), 4.00 (s, 6H), 3.06 (s, 3H),
2.50-1.90 (m, 8H), 1.50 (s, 9H); 13C NMR (100 MHz, CDCl3): δ 158.77, 155.52, 155.39, 153.75, 147.58,
145.98, 132.02, 128.97, 118.20, 106.68, 105.18, 100.73, 80.03, 56.73, 55.57, 45.27, 44.33, 28.94; MS
(MALDI-TOF, 2,5-dihydroxybenzoic acid as matrix): m/z (100%): 606.2369 [M+Na]+.
4.7.2. tert-Butyl exo-3-((6,7-dimethoxy-4-(4-(methylsulfonyl)phenylamino)quinazolin-2-
ylamino)methyl)-8-azabicyclo[3.2.1]octane-8-carboxylate (11b)
Following the general procedure, the title compound was prepared from compound 9 and 4-
o
1
methylsulfonylaniline in 65% yield; yellow solid; mp 156-158 C; H NMR (300 MHz, CDCl3): δ 8.00-
7.80 (m, 4H), 7.65 (s, 1H), 7.10 (s, 1H), 6.90 (s, 1H), 6.66 (brs, 1H), 4.25 (brs, 2H), 3.90 (m, 7H), 3.20 (m,
4H), 2.50 (m, 1H), 2.10-1.40 (m, 17H); MS (MALDI-TOF, 2,5-dihydroxybenzoic acid as matrix): m/z
(100%): 597.2533 M+.
4.7.3. (±)-4-(9-Benzyl-3,9-diazabicyclo[4.2.1]nonan-3-yl)-6,7-dimethoxy-N-(4-(methyl sulfonyl)phenyl)
quinazolin-2-amine (11c)
Following the general procedure, the title compound was prepared from compound 10 and 4-
methylsulfonylaniline in 54% yield; yellow solid; mp 171 oC; 1H NMR (300 MHz, CDCl3): δ 7.90 (m, 4H),
7.45-7.20 (m, 7H), 7.05 (s, 1H), 4.30 (s, 1H), 4.05-3.75 (m, 10H), 3.60-3.30 (m, 3H), 3.05 (s, 3H), 2.40-
1.50 (m, 6H).
4.7.4.
tert-Butyl
(±)-3-(6,7-dimethoxy-2-(4-(methylsulfonyl)phenylamino)quinazolin-4-yl)-3,9-
diazabicyclo[4.2.1]nonane-9-carboxylate (11d)
Reaction mixture of compound 11c (0.15 mmol) and 10% Pd/C (0.1 mmol) in dry MeOH (3 ml) was
hydrogenated for 5 hours at atmospheric pressure. The reaction mixture was filtered through celite and
concentrated under reduced pressure. To the solution of crude product in CH2Cl2 were added TEA (0.3
mmol) and Boc2O (0.2 mmol) and the reaction mixture was stirred for 2 hours. After extraction with
CH2Cl2, the combined organic layer was dried over anhydrous MgSO4, filtered, concentrated under reduced
pressure. Purification of the crude product by silica gel chromatography (CHCl3-MeOH 200:1) yielded
o
product as a yellow solid in 54% yield; mp 162 C; 1HNMR (300 MHz, CDCl3): δ 7.90 (m, 4H), 7.31 (s,
1H), 7.24 (d, J = 7.4 Hz, 1H), 7.06 (d, J = 7.4 Hz, 1H), 4.50-4.25 (m, 3H), 4.10-3.30 (m, 8H), 3.06 (s, 3H),
2.70-1.40 (m, 16H); MS (MALDI-TOF, α-cyano-4-hydroxycinnamic acid as matrix): m/z (100%):
584.2455 [M+H]+.
4.8. General procedure to generate 2-alkylamino-4-arylamino-6,7-dimethoxyquinazoline (12a-e)
Azabicyclic amine (7a-c, 1.2 mmol) and Hunig’s base (4.0 mmol) were added to a solution of compound (4
or 5, 1.0 mmol) in n-BuOH (2 ml) and the resulting mixture was stirred at reflux for 1 day. The reaction
mixture was cooled to room temperature, concentrated under reduced pressure, and the residue was purified
by column chromatography (CHCl3-MeOH 200:1) to obtain product.
- 9 -

4.8.1. tert-Butyl endo-3-(4-(2-fluoro-4-(methylsulfonyl)phenylamino)-6,7-dimethoxyquinazolin-2-
ylamino)-8-azabicyclo[3.2.1]octane-8-carboxylate (12a)
Following the general procedure, the title compound was prepared from azabicyclic amine 7a and
compound 5 in 14% yield; yellow solid; mp 178-181 oC; 1H NMR (300MHz, CDCl3): δ 8.87 (s, 1H), 7.82-
7.74 (m, 2H), 7.56 (brs, 1H), 6.94-6.91 (m, 2H), 4.27-4.25 (m, 4H), 4.01 (s, 3H), 4.00 (s, 3H), 3.11 (s, 3H),
2.36-2.31 (m, 3H), 1.90 (d, J = 4.5, 2H), 1.65 (brs, 3H), 1.50 (s, 9H); MS (MALDI-TOF, 2,5-
dihydroxybenzoic acid as matrix): m/z (100%): 602.2225 [M+H]+.
4.8.2. tert-Butyl exo-3-((6,7-dimethoxy-4-(4-(methylsulfonyl)phenylamino)quinazolin-2-
ylamino)methyl)-8-azabicyclo[3.2.1]octane-8-carboxylate (12b)
Following the general procedure, the title compound was prepared from azabicyclic amine 7b and
compound 4 in 27% yield; yellow solid; mp 181 oC; 1H NMR (300 MHz, CDCl3 + MeOD): δ 7.95 (m, 4H),
6.98 (s, 1H), 6.90 (s, 1H), 4.25 (brs, 2H), 4.00 (s, 6H), 3.35 (m, 2H), 3.09 (s, 3H), 2.20 (m, 1H), 2.00-1.40
(m, 17H); MS (MALDI-TOF, 2,5-dihydroxybenzoic acid as matrix): m/z (100%): 598.2514 [M+H]+.
4.8.3. tert-Butyl exo-3-((4-(2-fluoro-4-(methylsulfonyl)phenylamino)-6,7-dimethoxyquinazolin-2-
ylamino)methyl)-8-azabicyclo[3.2.1]octane-8-carboxylate (12c)
Following the general procedure, the title compound was prepared from azabicyclic amine 7b and
compound 5 in 43% yield; white solid; mp 220 oC; 1H NMR (300MHz, CDCl3): δ 8.93 (t, J = 8.2 HZ, 1H),
7.78-7.73 (m, 2H), 7.51 (s, 1H), 6.95 (s, 1H), 6.89 (s, 1H), 5.02 (brs, 1H), 4.29-4.22 (m, 2H), 4.01 (s, 6H),
3.37 (t, J = 6.1 Hz, 2H) , 3.10 (s, 3H), 2.26-2.18 (m, 1H), 1.96-1.95 (m, 2H), 1.71-1.45 (m, 6H), 1.47 (s,
9H); MS (MALDI-TOF, 2,5-dihydroxybenzoic acid as matrix): m/z (100%): 616.2614 [M+H]+.
4.8.4. (±)-2-(9-Benzyl-3,9-diazabicyclo[4.2.1]nonan-3-yl)-6,7-dimethoxy-N-(4-(methylsulfonyl)phenyl)
quinazolin-4-amine (12d)
Following the general procedure, the title compound was prepared from azabicyclic amine 7c and
o
1
compound 4 in 57% yield; yellow solid; mp 208 C; H NMR (300MHz, CDCl3): δ 7.92-7.87 (m, 2H),
7.84-7.79 (m, 2H), 7.55 (s, 1H), 7.42-7.40 (m, 2H), 7.34-7.21 (m, 3H), 7.02 (s, 1H), 6.93 (s, 1H), 4.52 (brs,
1H), 4.26 (brs, 1H), 3.97 (s, 6H), 3.77 (s, 2H), 3.74-3.62 (m, 1H), 3.39-3.31 (m, 3H), 3.07 (s, 3H), 2.02-
1.75 (m, 4H), 1.34-1.28 (m, 2H).
4.8.5. (±)-2-(9-Benzyl-3,9-diazabicyclo[4.2.1]nonan-3-yl)-N-(2-fluoro-4-(methylsulfonyl)phenyl)-6,7-
dimethoxyquinazolin-4-amine (12e)
Following the general procedure, the title compound was prepared from azabicyclic amine 7c and
compound 5 in 50% yield; yellow solid; mp 218 oC; 1H NMR (300MHz, CDCl3): δ 8.93 (s, 1H), 7.75-7.72
(d, 2H), 7.44-7.40 (m, 3H), 7.34-7.24 (m, 3H), 6.93 (s, 1H), 6.85 (s, 1H), 4.51 (brs, 1H), 4.23 (brs, 1 H),
4.00 (s, 3H), 3.99 (s, 3H), 3.78-3.68 (m, 3H), 3.41-3.35 (m, 3H), 3.08 (s, 3H), 2.22-2.10 (m, 1H), 2.05-1.95
(m, 2H), 1.80-1.75 (m, 2H), 1.37-1.26 (m, 1H); MS (MALDI-TOF, 2,5-dihydroxybenzoic acid as matrix):
m/z (100%): 592.2611 [M+H]+.
4.8.6.
tert-Butyl
(±)-3-(6,7-dimethoxy-4-(4-(methylsulfonyl)phenylamino)quinazolin-2-yl)-3,9-
diazabicyclo[4.2.1]nonane-9-carboxylate (12f)
The title compound was prepared from compound 12d according to the procedure for compound 11d.
o
Yield: 40%; yellow solid; mp 173 C; 1H NMR (300 MHz, CDCl3): δ 7.90 (m, 4H), 7.55 (s, 1H), 7.00 (s,
2H), 4.50-4.20 (m, 3H), 4.00 (s, 6H), 3.10 (m, 5H), 2.30-1.30 (m, 16H); MS (MALDI-TOF, α-cyano-4-
hydroxycinnamic acid as matrix): m/z (100%): 584.2572 [M+H]+.
4.8.7. tert-Butyl (±)-3-(4-(2-fluoro-4-(methylsulfonyl)phenylamino)-6,7-dimethoxyquinazolin-2-yl)-
3,9-diazabicyclo[4.2.1]nonane-9-carboxylate (12g)
The title compound was prepared from compound 12e according to the procedure for compound 11d.
o
1
Yield: 50%; yellow solid; mp 170-172 C; H NMR (300MHz, CDCl3): δ 8.92 (s, 1H), 7.80-7.73 (m, 2H),
7.47 (s, 1H), 6.93 (s, 1H), 6.86 (s, 1H), 4.46-4.23 (m, 3H), 4.00 (s, 6H), 3.37-3.33 (m, 1H), 3.10 (m, 4H),
2.17-2.15 (m, 2H), 1.97-1.84 (m, 3H), 1.49 (m, 11H); 13C NMR (100 MHz, CDCl3): δ 157.94, 155.58,
155.38, 153.69, 153.51, 152.52, 150.89, 146.33, 133.34, 124.21, 121.00, 114.29, 106.24, 103.15, 99.54,
79.29, 58.23, 56.37, 56.17, 54.05, 53.78, 45.58, 44.74, 31.79, 29.69, 28.53; MS (MALDI-TOF, 2,5-
dihydroxybenzoic acid as matrix): m/z (100%): 602.2386 [M+H]+.
- 10 -

4.9. Human GPR119 cAMP reporter assay
HEK293 cells (4x103 cells/well) were seeded on 96 half-well plates and incubated for 24h. The cells were
transected with GPR119 expression plasmid (OriGene Technologies, Inc., USA) using Lipofectamine and
Plus reagent (Life Technologies Corporation., USA). After 24h, transfected cells were incubated with
compounds dissolved in assay buffer (KRBH buffer containing 0.1% BSA and 500 μM 3-isobutyl-1-
o
methylxanthine) for 60 min at 37 C. Subsequently, cells were harvested with lysis buffer (50 mM
phosphate buffer containing 1 M KF and 1.25% Triton X-100, pH 7.0) for 10 min at room temperature and
the assay was performed using the cAMP homogeneous time-resolved fluorescence kit (CIS bio
international, France).
Acknowledgments
The authors thank to Dr. Moon Ho Son (Dong-A Pharm. Co. Ltd.) for hGPR119 cAMP reporter assay. We
also thank to Pharmacal Research Institute and Central Laboratory of Kangwon National University for the
use of analytical instruments and bioassay facilities.
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*Graphical Abstract (for review)
R
O
O
N
R
Boc
N
NH
N
H
N
N
NH
F
N
Boc
N
Boc
12a
12c
H3CO2S
12g
human GPR119 agonists
EC50 = 1 M