Synthesis and biological evaluation of thienopyrimidine derivatives as GPR119 agonists

Article abstract of DOI:10.1016/j.bmcl.2014.07.020

A series of thienopyrimidine derivatives was synthesized and evaluated for their GPR119 agonistic ability. Several thienopyrimidine derivatives containing R¹ and R² substituents displayed potent GPR119 agonistic activity. Among them, compound 5d, which is a prototype, showed good in vitro activity with an EC₅₀ value of 3 nM and human and rat liver microsomal stability. Compound 5d exhibited no CYP inhibition and induction, Herg binding, or mutagenic potential. Compound 5d showed increase insulin secretion in beta TC-6 cell and lowered the glucose excursion in mice in an oral glucose-tolerance test.

Full text of DOI:10.1016/j.bmcl.2014.07.020

Bioorganic & Medicinal Chemistry Letters xxx (2014) xxx–xxx
Contents lists available at ScienceDirect
Bioorganic & Medicinal Chemistry Letters
journal homepage: www.elsevier.com/locate/bmcl
Synthesis and biological evaluation of thienopyrimidine derivatives
as GPR119 agonists
Moon-Kook Jeon a, , Kyu Myung Lee a, , Il Hyang Kim a, Yoon Kyung Jang a,b, Seung Kyu Kang a, Jun Mi Lee a,
Kwan-Young Jung a, Jaladi Ashok Kumar a, Sang Dal Rhee a, Won Hoon Jung a, Jin Sook Song a,
Myung Ae Bae a, Kwang Rok Kim a, Jin Hee Ahn a,b,
⇑
a Drug Discovery Division, Korea Research Institute of Chemical Technology, Republic of Korea
b Department of Medicinal and Pharmaceutical Chemistry, University of Science and Technology, 305-333, Republic of Korea
a r t i c l e i n f o
a b s t r a c t
Article history:
A series of thienopyrimidine derivatives was synthesized and evaluated for their GPR119 agonistic
ability. Several thienopyrimidine derivatives containing R1 and R2 substituents displayed potent
GPR119 agonistic activity. Among them, compound 5d, which is a prototype, showed good in vitro activ-
ity with an EC50 value of 3 nM and human and rat liver microsomal stability. Compound 5d exhibited no
CYP inhibition and induction, Herg binding, or mutagenic potential. Compound 5d showed increase insu-
lin secretion in beta TC-6 cell and lowered the glucose excursion in mice in an oral glucose-tolerance test.
Ó 2014 Elsevier Ltd. All rights reserved.
Received 3 June 2014
Revised 6 July 2014
Accepted 8 July 2014
Available online xxxx
Keywords:
Diabetes
Thienopyrimidine
GPR119
Treatment
Diabetes is a serious metabolic disorder that occurs when the
pancreas does not produce enough insulin, or the body cannot
effectively use existing insulin.1 Hyperglycemia (high blood
glucose) can lead to various health consequences such as kidney
damage, heart disease, stroke, nerve damage and blindness. Type
2 diabetes mellitus (T2DM, or noninsulin dependent), is the most
common form of diabetes caused by insulin resistance, and loss
of pancreatic b-cell function and approximately 95% diabetic
patients are suffering from type 2 diabetes. This health burden is
growing at an alarming rate, and it is estimated that there are
approximately 350 million diabetic people globally. The preva-
lence of the disease is expected to escalate to 439 million by
MeO2S
O
O
O
N
N
N
O
O
N
N
O
O
N
N
N
N
N
N
F
F
N
O
O
SO2Me
SO2Me
Figure 1. Representative bicyclic GPR119 agonists.
2030.2–4 Although,
a variety of treatments are available for
T2DM, many patients are unable to achieve their target HbA1c
level.5 Considerable attention has been focused on overcoming this
public health challenge worldwide. Hence, there is a strong need
for novel approaches to achieve better glycemic control and nor-
moglycemia. Strategies that promote significant glycemic control
by limiting hypoglycemia and cardiovascular side effects by
enhancing insulin secretion in a glucose dependent manner could
offer robust treatment for T2DM.
GPR119 is a member of the class A G protein-coupled receptor
(GPCR) family, and it is highly expressed in pancreatic b-cells
and intestinal endocrine cells.6–8 Upon activation by its endoge-
nous ligand, intracellular cAMP accumulates and adenylate cyclase
activation enhances the effect of glucose-stimulated insulin secre-
tion (GSIS) and GLP-1 release. Thus GPR119 represents a promising
target for the treatment of type 2 diabetes and obesity owing to its
ability to improve glucose homoeostasis while concurrently
slowing gastric emptying, reducing food intake and promoting
weight loss.9,10
Endogenous ligands for GPR-119 have been identified including
lysophosphatidylcholine (LPC) and oleoylethanolamide (OEA).9–11
Moreover, numerous small molecule GPR119 agonists have been
⇑
Corresponding author.
E-mail address: jhahn@krict.re.kr (J.H. Ahn).
These authors contributed equally to this work.
http://dx.doi.org/10.1016/j.bmcl.2014.07.020
0960-894X/Ó 2014 Elsevier Ltd. All rights reserved.
Please cite this article in press as: Jeon, M.-K.; et al. Bioorg. Med. Chem. Lett. (2014), http://dx.doi.org/10.1016/j.bmcl.2014.07.020

2
M.-K. Jeon et al. / Bioorg. Med. Chem. Lett. xxx (2014) xxx–xxx
R1
N
identified in recent years. Among them, structurally rigid bicyclic
compounds were identified as promising scaffolds. These bicyclic
analogues exhibited potent GPR119 agonistic activity, efficacy
and PK profiles (Fig. 1).12–16
S
N
R2
This prompted us to look for an alternate bicyclic scaffold, as a
result, we identified the thienopyrimidine scaffold.17 In this pres-
ent work we wish to report the synthesis and biological evaluation
of thienopyrimidine derivatives as GPR119 agonists (Fig. 2).
The general method for compound synthesis is outlined in
Scheme 1. As shown in Scheme 1, commercially available Boc
protected piperidine derivative 1 was coupled with 6-bromo-
4-chlorothieno[3,2-d]pyrimidine to yield coupled product 2, which
was treated with diverse aryl boronic acids by Suzuki coupling in
the presence of palladium catalyst afforded compound 5 with good
yield. Meanwhile, Boc-protected piperidinone 3 was converted to
4-methylaminopiperidine derivative 4 via reductive amination,
and it was then coupled with 6-bromo-4-chlorothieno[3,2-
d]pyrimidine, followed by Suzuki reaction to yield compound 5.
Deprotection of compound 5 with 4 M HCl afforded compound 6,
which was derivatized by diverse electrophiles to give the final
thienopyrimide derivative 7.
Figure 2. Structure of thienopyrimidine scaffold.
O
N
1
O
O
a
N
O
A
A = OHor NH
2
X
N
S
N
Br
2
a
O
3
O
4
b
N
O
N
O
O
N
H
c
Thus synthesized thienopyrimidine derivatives were evaluated
in vitro for GPR119 agonistic activity, and the results are summa-
rized in Tables 1–3. First, we fixed the 4-methylsulfonylphenyl
substituent at the R2 position on the thienopyrimidine ring, and
derivatized at the R1 position. As shown in Table 1, 5a and 7a
showed weak agonistic activities; however, the introduction of a
Boc-protected N-methylpiperidine to thienopyrimidine 5b exhib-
ited good in vitro activity with an EC50 value of 39 nM. Also, pyrim-
idine substituted N-methylpiperidine derivative 7b displayed
moderate potency (EC50 = 1200 nM). 4-Oxypiperidine derivatives
5c and 7c also activated GPR119 with EC50 values of 100 nM and
240 nM, respectively.
O
R1
N
O
N
NH
X
X
X
d
e
S
S
S
N
N
N
N
N
N
6
Ar
Ar
Ar
5
7
Scheme 1. Reagents and conditions: (a) 6-Bromo-4-chlorothieno[3,2-d]pyrimidine,
A = OH, NaH, THF room temperature, 12 h; A = NH2, DMSO, room temperature, 12 h;
(b) 40% methylamine in methanol, NaBH(OAc)3, 1,2-dichloroethane, room temper-
ature, 12 h; (c) Pd(PPh3)4, Na2CO3, arylboronic acid, 1,4-dioxane, 110 °C, 24 h; (d)
4 M HCl in dioxane, room temperature, 2 h; (e) carbamate, alkylchloroformate,
triethylamine, CH2Cl2, room temperature, 1 h; amide: acylhalide, triethylamine,
CH2Cl2, DMF, room temperature, 12 h; pyrimidine; 2-chloro-5-ethylpyrimidine,
triethylamine, DMF, 130 °C, 3 h.
Based on the data shown in Table 1, we further derivatized the
R1 position with an N-methylaminopiperidine group. As shown in
Table 1
In vitro GPR119 agonist activity of thienopyrimidine derivatives
Compound
Structure
% Activation at 1
49
l
Ma
Human EC50 (nM)
NDb
S
S
H
N
O
S
O
H3C
5a
N
N
N
N
N
O
CH3
CH3
O
CH3
H
N
O
S
O
H3C
7a
5b
7b
5c
7c
49
76
67
64
75
ND
N
N
N
C2H5
CH3
N
S
O
S
O
H3C
39
N
N
N
O
CH3
CH3
O
N
CH3
S
CH3
N
O
S
O
H3C
1200
100
240
N
N
N
N
C2H5
S
S
O
O
O
S
O
H3C
N
N
N
N
N
O
CH3
CH3
O
CH3
O
S
O
H3C
N
N
N
C2H5
a
Activation relative to GSK1292263.
Not determined.
b
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M.-K. Jeon et al. / Bioorg. Med. Chem. Lett. xxx (2014) xxx–xxx
3
Table 2
In vitro GPR119 agonist activity of 4-methylaminopiperdinothienopyrimidine derivatives
Compound
Structure
% Activation at 1
76
l
Ma
Human EC50 (nM)
CH3
N
S
O
S
O
H3C
5b
39
N
N
N
O
CH3
CH3
O
CH3
S
CH3
N
O
S
7d
7e
67
72
82
N
N
N
O
CH3
CH3
H3C
O
O
S
CH3
N
O
S
O
H3C
173
N
N
N
O
CH3
N
S
O
S
O
O
H3C
7f
60
49
781
NDb
N
N
N
CH3
H3C
CH3
N
S
O
S
O
H3C
H
N
7g
N
N
N
CH3
CH3
O
CH3
N
S
O
S
H3C
O
S
7h
70
1200
N
N
N
O
O
H3C
CH3
a
Activation relative to GSK1292263.
Not determined.
b
Table 3
In vitro GPR119 agonist activity of 4-methylaminopiperdinothienopyrimidine derivatives
Compound
Structure
% Activation at 1
l
Ma
Human EC50 (nM)
F
S
CH3
N
O
S
5d
94
3
H3C
O
N
N
N
O
CH3
CH3
O H3C
F
CH3
S
O
N
S
7i
7j
45
ND
26
H3C
O
N
N
N
O
CH3
CH3
O H3C
F
F
S
CH3
N
O
S
O
105
H3C
N
N
N
O
CH3
O
S
CH3
N
O
CH3
S
O
7k
7l
115
65
140
NDb
ND
H3C
N
N
N
CH3
O
F
S
CH3
N
O
S
H3C
N
N
N
O
O
F
S
CH3
N
O
S
H3C
7m
>50
CH3
CH3
O
N
N
N
N
O
N
a
Activation relative to GSK1292263.
Not determined.
b
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4
M.-K. Jeon et al. / Bioorg. Med. Chem. Lett. xxx (2014) xxx–xxx
Table 4
Table 2, isopropyl carbamate 7d was also active with an EC50 value
of 82 nM. Amide derivatives (7e and 7f) were weaker than carba-
mate derivatives (5b and 7d). Introduction of an isopropyl urea
moiety resulted in loss of activity (7g). Also, isopropyl sulfonamide
(7h) showed moderated GPR119 agonistic activity.
Stability, CYP, hERG, AMES and cytotoxicity of compound 5d
Assay
Results
Liver microsomal stability
(Human)
>95% parent was remained after 30 min
incubation
Liver microsomal stability
(rat)
CYP inhibition
>95% parent was remained after 30 min
incubation
1A2: no inhibition
2A6: no inhibition
2C19: no inhibition
Diverse substituted aryl and heteroaryl groups were introduced
at the R2 position of the thienopyrimidine ring. As expected, meth-
ylsulfonyl substituted aryl or heteroaryl derivatives showed good
in vitro activity (data not shown). Therefore, we focused on R2
modification with 2-fluoro-4-methylsulfonylphenyl group, and
the results were summarized in Table 3. tert-Butyl and isopropyl
carbamate (5d) showed good in vitro potency with EC50 value of
3 nM. However, the introduction of an ethyl group resulted in loss
of activity (compound 7i). Methylcyclopropyl carbamate (7j)
exhibited good activity (EC50 = 26 nM), whereas other amides (7k
and 7l) and oxadiazole (7m) resulted in a loss of activity.
From the results of our in vitro data, 5d was selected as a
prototype compound. Next, compound 5d was investigated for
its stability, ability to induce/inhibit CYP, Herg binding and cyto-
toxicity. As shown in Table 4, compound 5d is metabolically stable
in human and rat liver microsomes, with over 95% of the parent
compound remaining after 30 min incubation. In CYP inhibition/
induction assays with several CYP subtypes, compound 5d did
not significantly inhibit or induce CYP. Compound 5d showed no
2C9: IC50 = 76.85 lM
3A4: no inhibition
2D6: no inhibition
2B6: no inhibition
CYP induction
1A2: no induction
2B6: no induction
2C9: no induction
2E1: no induction
3A4: no induction
Herg
Mini AMES
13% at 10 lM (binding assay)
TA100: no mutation
TA98: no mutation
NIH 3T3: IC50 = 49.77 lM
Cytotoxicity
L929: IC50 = 65.79
HFL-1: IC50 >100
CHO-K1: IC50 = 53.37
lM
lM
l
M
Herg binding (13% inhibition at 10
in the AMES assay.
lM) and mutagenic potential
To evaluate that 5d has direct effects on beta cells, insulin secre-
tion was measured in the pancreatic beta cell line, TC-6. As can be
seen in Figure 3, exposure to 5d at concentrations ranging from
10 nM to 100 nM increased glucose-stimulated insulin secretion
(GSIS) in a dose-dependent manner in TC-6 cells.
To evaluate in vivo efficacy, oral glucose tolerance tests (OGTT)
were performed using compound 5d. Plasma glucose levels were
determined based on the AUC of the glucose concentration, and
they were significantly reduced at 15 mg/kg dose (Fig. 4).
In conclusion, we identified a series of thienopyrimidine deriv-
atives as GPR119 agonists. Several thienopyrimidine derivatives
with R1 and R2 substituents were found to be potent GPR119
agonists. Among them, compound 5d was the most active with
an EC50 value of 3 nM and showed good human and rat liver micro-
somal stability. Compound 5d exhibited no CYP inhibition and
induction, hERG binding, or mutagenic potential. Compound 5d
induced increased insulin secretion from beta cell and reduced
the AUC of glucose in vivo OGTT. We are currently producing
further modification of this prototype that will be examined in
the near future.
Figure 3. Glucose stimulated insulin secretion in TC-6 cells. ⁄P <0.05.
Figure 4. In vivo oral glucose tolerance test of compound 5d in normal mice. Results are expressed as means SEM for n = 7 mice/group. ⁄P <0.05, ⁄⁄P <0.01.
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M.-K. Jeon et al. / Bioorg. Med. Chem. Lett. xxx (2014) xxx–xxx
5
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Supplementary data
Supplementary data associated with this article can be found,
in the online version, at http://dx.doi.org/10.1016/j.bmcl.2014.07.
020.
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