C5-Alkyl-2-methylurea-substituted pyridines as a new class of glucokinase activators

Article abstract of DOI:10.1021/ml500341w

Glucokinase (GK) activators represent a class of type 2 diabetes therapeutics actively pursued due to the central role that GK plays in regulating glucose homeostasis. Herein we report a novel C5-alkyl-2-methylurea-substituted pyridine series of GK activators derived from our previously reported thiazolylamino pyridine series. Our efforts in optimizing potency, enzyme kinetic properties, and metabolic stability led to the identification of compound 26 (AM-9514). This analogue showed a favorable combination of in vitro potency, enzyme kinetic properties, acceptable pharmacokinetic profiles in preclinical species, and robust efficacy in a rodent PD model.

Full text of DOI:10.1021/ml500341w

Letter
pubs.acs.org/acsmedchemlett
C5-Alkyl-2-methylurea-Substituted Pyridines as a New Class
of Glucokinase Activators
Xiaohui Du,*,† Ronald J. Hinklin,§ Yumei Xiong,† Paul Dransfield,† Jaehyeon Park,† Todd J. Kohn,†
Vatee Pattaropong,† SuJen Lai,† Zice Fu,† Xianyun Jiao,† David Chow,† Lixia Jin,† Jasmine Davda,†
Murielle M. Veniant,‡ Deborah A. Anderson,§ Brian R. Baer,§ Josef R. Bencsik,§ Steven A. Boyd,§
Mark Joseph Chicarelli,§ Peter J. Mohr,§ Bin Wang,§ Kevin R. Condroski,§ Walter E. DeWolf,§
Marion Conn,† Thanhvien Tran,† Jerry Yang,† Thomas D. Aicher,§ Julio C. Medina,† Peter Coward,†
and Jonathan B. Houze†
†Amgen, Inc., 1120 Veterans Boulevard, South San Francisco, California 94080, United States
‡Amgen, Inc., One Amgen Center Drive, Thousand Oaks, California 91320, United States
§Array BioPharma, 3200 Walnut Street, Boulder, Colorado 80301, United States
S
* Supporting Information
ABSTRACT: Glucokinase (GK) activators represent a class of type 2
diabetes therapeutics actively pursued due to the central role that
GK plays in regulating glucose homeostasis. Herein we report a novel
C5-alkyl-2-methylurea-substituted pyridine series of GK activators
derived from our previously reported thiazolylamino pyridine series.
Our efforts in optimizing potency, enzyme kinetic properties, and
metabolic stability led to the identification of compound 26 (AM-9514).
This analogue showed a favorable combination of in vitro potency,
enzyme kinetic properties, acceptable pharmacokinetic profiles in
preclinical species, and robust efficacy in a rodent PD model.
KEYWORDS: Type 2 diabetes, glucokinase activator, GKA, methyl urea-substituted pyridines
ype 2 diabetes is a chronic metabolic disease affecting
advantage in the treatment of type 2 diabetes.8 To date, a number
of GK activators have advanced into clinical trials for the
treatment of type 2 diabetes.9−11
T
over 300 million people worldwide.1 It is characterized
by elevated fasting plasma and hepatic glucose production and
insulin resistance. It is a major risk factor for both microvascular
and macrovascular diseases.2−4 Despite the availability of multiple
antihyperglycemic agents, many patients still remain unable to
safely achieve and maintain tight glycemic control, underscoring
the need for more effective therapies.
Glucokinase (GK) is a member of the hexokinase family of
intracellular enzymes. It is responsible for the conversion of
glucose to glucose-6-phosphate, the first step in glucose utiliza-
tion.5 GK is present in both pancreatic β-cells and liver hepatic cells.
In the pancreas, it serves as a glucose sensor to control glucose-
stimulated insulin secretion, while hepatic GK plays an important
role in regulating net glucose utilization and production.6
It has been demonstrated that glucokinase activators (GKAs)
can influence the enzyme’s kinetic profile by modulating both Km
for glucose (also known as S0.5) and Vmax (maximal velocity of the
glucose phosphorylation reaction catalyzed by GK).7 As a result,
GKAs control blood glucose concentrations by enhancing
the ability of pancreatic β-cells to sense glucose and increase
insulin secretion in a glucose-dependent manner. In addition,
GKAs act in the liver to improve glucose disposal by increasing
glycogen storage and decreasing hepatic glucose output. The
possible synergistic effect of this dual mechanism may provide an
We have previously disclosed a series of thiadiazolylamino
pyridine GKAs represented by compound A.12,13 To identify
GKAs that are structurally distinct from that, we pursued a novel
methyl urea-substituted pyridine series, where the critical
bidentate hydrogen bond to Arg63 was maintained by the methyl
urea moiety.14 Compound A had thioether substitutions at C5
of the middle pyridine ring. In pursuing more structure diversity,
we would like to replace the thioether in compound A with a
carbon-based linker in compound B as shown in Figure 1.
Hypoglycemia is a key dose limiting adverse effect of
glucokinase activators.15−17 In an attempt to mitigate this risk,
we targeted GKAs that would not lower the S0.5 below a certain
limit nor raise Vmax above a certain limit, preferably with S0.5
>
0.6 mM and Vmax between 0.8 and 1.3 of unactivated GK.
A similar strategy has been proposed by others.10,11,18 Our
threshold values were chosen in part based on data with Piragliatin
(S0.5 of 0.28 mM and Vmax of 1.7) and MK-0941 (S0.5 of 0.25 mM
Received: August 22, 2014
Accepted: November 22, 2014
© XXXX American Chemical Society
A
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Letter
rate in both rat and human liver microsomes. The microsomal
stability of this series was improved by the incorporation of a
variety of polar functional groups such as alcohols and ethers into
the structure (compounds 2 and 4−7). For the ethers, potency
improved when elongating the linker from two carbon to three
and four (2, 3, and 4). However, the Vmax values for these
compounds were lower than the target range. For the alcohols,
although they were less potent than their ether analogues (2 vs 5;
4 vs 6), one of the primary alcohols, 6, had acceptable potency,
kinetic parameters, and microsomal stability. However, com-
pound 6 had poor stability in primary human hepatocytes (88%
was metabolized).22 To reduce the hepatic clearance, the tertiary
alcohol 7 was made. Although this improved the stability in
primary human hepatocytes (28% metabolized),22 it also reduced
the Vmax to an unacceptable level (0.73).
We then explored cyclic moieties with one carbon linker such
as cyclohexyl methyl or THP methyl substitution. The Vmax
values for these compounds (8 and 9) were also low.
Because of the suboptimal Vmax associated with 3-O-ethyl-
pyridine substitution, we changed the substitution to 3-O-
ethylpyrazole. Table 2 shows that the analogues with
Figure 1. From thioether-substituted thiazolylamino pyridines (A) to
methyl urea-substituted pyridines (B).
and Vmax of 1.3),19 developed by Roche and Merck, respectively.
Both compounds were associated with increased risk of
hypoglycemia in clinical trials.16,20 In addition, compounds with
Vmax lower than 0.8 could result in the phosphorylation rate of the
enzyme-activator complex to be lower than the native enzyme at
glucose concentrations obtainable in the fed state.21 Therefore, we
targeted GKAs with Vmax between 0.8 and 1.3.
A number of C5-alkyl substituted analogues with 3-O-
ethylpyridine substitution are summarized in Table 1. The simple
isobutyl substituted analogue 1 was potent (EC50 = 0.12 μM) but
metabolically unstable, indicated by the relatively high turnover
Table 2. Exploration of C5 Substitution of GKAs with
3-O-Pyrazole
Table 1. Exploration of C5 Substitution of GKAs with
3-O-Ethylpyridine
a
Mean standard deviations for the EC50, S0.5, and Vmax are 30%, 17%,
and 4%, respectively (n = 2). Also, human recombinant GK was used
b
in all the assays. For the EC50 assay, the glucose concentration was
c
fixed at 5 mM. Percent turnover: percentage of parent compound
being metabolized after incubating in 0.25 mg/mL of rat or human
liver microsomes for 30 min at 37 °C with an initial parent con-
d
centration of 1 μM. Maximal velocity of the glucose phosphorylation
reaction catalyzed by activated GK/unactivated GK.
3-O-ethylpyrazole had improved Vmax values compared to the
3-O-ethylpyridines, although the potency was reduced somewhat
(10 vs 4; 11 vs 7; 12 vs 9). These compounds generally had good
microsomal stability.
Compound 12 had acceptable potency, kinetic parameters,
and good stability in microsomes and primary human hepato-
cytes (15% metabolized).22 Furthermore, it has a low molecular
weight (359) and very good solubility (718 μg/mL at pH 7.4,
PBS buffer). Table 3 shows the pharmacokinetic properties of 12
in mice and rats. Although it had modest clearance in both rat
and mouse, it had excellent bioavailability in mouse (81%). In
addition, we studied the interspecies differences in the kinetic
properties of compound 12−GK complex. Compound 12 had a
a
Mean standard deviations for the EC50, S0.5, and Vmax are 30%, 17%,
and 4%, respectively (n = 2). Also, human recombinant GK was used
in all the assays. For the EC50 assay, the glucose concentration was
fixed at 5 mM. Percent turnover: percentage of parent compound
being metabolized after incubating in 0.25 mg/mL of rat or human
liver microsomes for 30 min at 37 °C with an initial parent
concentration of 1 μM. Maximal velocity of the glucose
phosphorylation reaction catalyzed by activated GK/unactivated GK.
b
c
d
B
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Table 3. Pharmacokinetics Properties of 12
Table 4. Modifications of the THP Group at C5
a
a
IV
PO
CL
(L/h/kg)
Vss
(L/kg)
T1/2
(h)
F
compd species
(%) AUC (μM·h)
12
12
rat
1.5
1.7
0.8
1.5
0.41
0.72
19
81
3.51
13.3
mouse
a
Rat and mouse: IV dose 1 mg/kg, n = 3; PO dose 10 mg/kg, n = 3.
Vmax of 1.12 and a S0.5 of 0.82 in mouse. Although the difference
in the Vmax between human and mouse was significant, it was
within our target range. Compound 12 (AM-0822) was chosen
to proceed into in vivo studies to verify its efficacy.
Compound 12 (AM-0822) was tested in a dose−response oral
glucose tolerance test (OGTT) using C57Bl/6 mice23 (Figure 2)
a
Mean standard deviations for the EC50, S0.5, and Vmax are 30%, 17%,
and 4%, respectively (n = 2). Also, human recombinant GK was used
b
in all the assays. For the EC50 assay, the glucose concentration was
c
fixed at 5 mM. Maximal velocity of the glucose phosphorylation
reaction catalyzed by activated GK/unactivated GK.
Figure 2. OGTT data for 12 (AM-0822) in C57Bl/6 mice. Statistical
significance compared to vehicle treatment is denoted by *(p < 0.05),
**(p < 0.01), ***(p < 0.001), and ****(p < 0.0001), as determined by
two-way ANOVA, and is color-coded to the treatment in the figure
legends.
Table 5. Modifications on the 3-O-Pyrazole
and showed efficacy at 3 mg/kg, the lowest dose administered.
The area-under-the-curve (AUC) for the blood glucose was
decreasedby15% at 3 mg/kg and 32% at 10mg/kg. The 10mg/kg
dose was equally efficacious as the 30 mg/kg dose. The plasma
glucose level stabilized at 77 mg/dL between 1.5 and 2 h for the
30 mg/kg dose, well above the hypoglycemic range (<50 mg/dL),
thus confirming our target range for the kinetic parameters on GK
induced by our GKA.
However, compound 12 had significant brain penetration in
rat. The unbound concentrations in the brain were 29% of
unbound concentrations in plasma in rat. Because of the fact that
GK is also expressed in brain, there will be risk of interfering with
counter regulation once GKA penetrates the brain and lowers the
brain blood glucose level. Therefore, this was considered to be an
undesirable attribute in a potential GKA therapeutic. We then
continued to seek GKAs with less brain penetration (less than
10% is desired) in addition to a similar or better overall profile
than 12.
a
Mean standard deviations for the EC50, S0.5, and Vmax are 30%, 17%,
Our subsequent efforts were centered on modifications of 12,
and Table 4 shows some of the analogues explored. Hydroxyl
group substitution at the benzylic position (13), morpholino
replacement of THP (14), and alpha substitution to the oxygen
of THP (15) all led to compounds with decreased potency.
The only compound with similar potency (16) displayed a Vmax
less than 0.8.
and 4%, respectively (n = 2). Also, human recombinant GK was used
b
in all the assays. For the EC50 assay, the glucose concentration was
c
fixed at 5 mM. Maximal velocity of the glucose phosphorylation
reaction catalyzed by activated GK/unactivated GK.
We then investigated cyclic alcohols at C5 (Table 6).
Secondary or tertiary hydroxyl would prevent secondary
metabolism as shown in 7. Cyclic compounds would also reduce
the degrees of freedom in the molecule, which might improve
Table 5 shows modifications on the pyrazole ring. These
analogues (17 and 18) are also less potent than 12.
C
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Table 6. Cyclic Alcohols at C5
Figure 3. In vitro parameters of 26: 4% HSA EC50 (the EC50 measured in
the presence of 4% human serum albumin (HSA)).
Table 7. Pharmacokinetic Properties of 26
IV
PO
(%) AUC (μM·h)
CL
(L/h/kg)
Vss
(L/kg)
T1/2
(h)
F
compd species
a
26
26
26
rat
1.6
0.8
1.2
2.2
2.0
1.2
5.5
9
0.39
6.84
6.4
b
mouse
0.91
0.64
44
72
a
dog
a
Rat and dog: IV dose 0.5 mg/kg, n = 3; PO dose 2.0 mg/kg, n = 3.
Mouse: IV dose 1.0 mg/kg, n = 3; PO dose 5.0 mg/kg, n = 3.
b
a
Scheme 1. Synthesis of 26
a
a
Reagents and conditions: (a) BnBr, K2CO3, CH3CN, 93%; (b)
Mean standard deviations for the EC50, S0.5, and Vmax are 30%, 17%,
Tebbe reagent, THF, 58%; (c) H2O2, CCl3CN, DCM, separate the
trans/cis isomers, 48%; (d) LiAlH4, THF, 90%; (e) Ac2O, pyridine,
DCM, 94%; (f) TBSOTf, DIEPA, DCM, 87%; (g) K2CO3, MeOH,
94%; (h) iodobenzene diacetate, TEMPO, DCM, 85%; (i) 1-(5-
bromo-3-((1-ethyl-1H-pyrazol-5-yl)oxy)pyridin-2-yl)-3-methylurea,
MeLi, BuLi, aldehyde, THF; then MeSO3H, MeOH, 57% overall yield;
(j) MeSO3H, 10% Pd/C, H2, MeOH, 59%.
and 4%, respectively (n = 2). Also, human recombinant GK was used
b
in all the assays. For the EC50 assay, the glucose concentration was
c
fixed at 5 mM. Maximal velocity of the glucose phosphorylation
reaction catalyzed by activated GK/unactivated GK.
pharmacokinetic properties.24 Another advantage of alcohols
is that they generally have increased polarity and polar surface
area than ethers, which might lower the brain penetration level.25
The cyclohexanols (19−22) were more potent than 12, but most
of them had lower Vmax with the exception of compound 22;
however, for 22, the S0.5 was out of the desired range.
bioavailability in dog. It was scaled up for further in vivo studies.
The synthesis is shown in Scheme 1.
The synthesis of 26 started from 3-oxocyclobutanecarboxylic
acid. It was protected with a benzyl group and then converted
to benzyl 3-methylenecyclobutanecarboxylate 26.2 through the
Tebbe olefination. Epoxidation of the double bond in 26.2 and
isolation of the less polar isomer led to 26.3, which was reduced
to diol 26.4 with lithium aluminum hydride. The primary hydroxyl
group in 26.4 was selectively protected with an acetate group
(26.5), and the tertiary hydroxyl group was protected with a TBS
group (26.6). Deprotection of the acetate group in 26.6 freed
the primary hydroxyl group for further functional group
transformations. Compound 26.7 was oxidized to aldehyde 26.8
under mild conditions. Lithiation of 1-(5-bromo-3-((1-ethyl-1H-
pyrazol-5-yl)oxy)pyridin-2-yl)-3-methylurea26 and addition of the
organolithium generated to aldehyde 26.8 followed by subsequent
deprotection of the TBS protection group resulted in diol 26.9.
The Vmax parameters in the cyclobutanols were more
encouraging. Both cis and trans isomers (23 and 24) had higher
Vmax values than 12, with one of the isomers, 24, having a Vmax of
0.96. To improve their potency to a more desirable range, a
methyl group was added to form the tertiary cyclobutyl alcohols.
One of the isomers, 26, had a Vmax of 0.91 and an EC50 of
0.29 μM, both improved over 12. The S0.5 decreased to 0.68 mM
but was still within our target range.
Compound 26 (AM-9514) had excellent solubility, perme-
ability, in vitro clearances and free fractions in plasma protein
binding (Figure 3). It had desirable pharmacokinetics in
mouse and dog with a clearance of 0.91 L/h/kg in mouse
and 0.64 L/h/kg in dog (Table 7). It also had excellent oral
D
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The benzylic hydroxyl group in 26.9 was removed through hydro-
genation under strong acidic conditions to provide compound 26.
The cis relationship of the methyl group and the benzylic methyl
group relative to the cyclobutane ring in 26 was confirmed by
2D-NMR experiments.
Before further in vivo study, we measured the kinetic properties
of 26 on mouse GK (Vmax = 1.14 and S0.5 = 0.69) and found them
within our target range and also closer to human parameters
compared to compound 12. The antidiabetic efficacy of 26 was
tested in an OGTT at doses of 0.3, 1, 3, and 10 mg/kg in fasted
ob/ob mice (Figure 4). It showed dose-proportional lowering
of blood glucose, with a 43% AUC decrease at 10 mg/kg.
maintained above 85 mg/dL at all time points for both 10 and
30 mg/kg doses, well above its efficacious dose of 1 mg/kg.
Therefore, 26 showed no hypoglycemia at the doses tested.
Similar results of normoglycemic glucose lowering test were
also obtained in rat of compound 26. In addition, compound 26
showed low brain penetration in rat. The unbound concentrations
in the brain were 4% of unbound plasma concentrations in rat.
In conclusion, a new class of C5-alkyl-2-methylurea-substituted
pyridines as glucokinase activators was developed based on
our previous thiazolylamino pyridine GKAs. We optimized
the potency and kinetic parameters of this class and identified
compound 26 (AM-9514) with good potency, desirable GK
kinetic parameters, and favorable in vitro and in vivo profiles
including pharmacokinetic properties.
ASSOCIATED CONTENT
* Supporting Information
■
S
Assay, in vivo procedures, experimental procedures, and data for
all compounds. This material is available free of charge via the
Internet at http://pubs.acs.org.
AUTHOR INFORMATION
Corresponding Author
*E-mail xdxiaohui@gmail.com.
■
Notes
The authors declare no competing financial interest.
REFERENCES
■
Figure 4. OGTT data of 26 (AM-9514) in ob/ob mice. Compound 26
lowered blood glucose in a dose-dependent manner. Statistical
significance compared to vehicle treatment is denoted by *(p < 0.05),
**(p < 0.01), ***(p < 0.001), and ****(p < 0.0001), as determined by
two-way ANOVA, and is color-coded to the treatment in the figure
legends.
(1) Danaei, G.; Finucane, M. M.; Lu, Y.; Singh, G. M.; Cowan, M. J.;
Paciorek, C. J.; Lin, J. K.; Farzdfar, F.; Khang, Y.-H.; Stevens, G. A.; Rao,
M.; Ali, M. K.; Riley, L. M.; Robinson, C. A.; Ezzati, M. National,
regional, and global trends in fasting plasma glucose and diabetes
prevalence since 1980: systemic analysis of health examination surveys
and epidemiological studies with 370 country-years and 2.7 million
participants. Lancet 2011, 378, 31−40.
(2) Nazimek-Siewniak, B.; Moczulski, D.; Grzeszczak, W. Risk of
macrovascular and microvascular complications in Type 2 diabetes:
results of longitudinal study design. J. Diabetes Complications 2002, 16,
271−276.
Compound 26 was further tested in a normoglycemic
glucose lowering test to evaluate its hypoglycemic risk. Fed
male C57Bl/6 mice were orally dosed with vehicle or 26 without
glucose challenge and then food was removed for 4 h. Blood
glucose levels were measured at various time points (Figure 5).
The elevation in glucose seen with the vehicle is presumably the
result of handling stress on animals. Blood glucose levels were
(3) Kles, K. A.; Vinik, A. I. Pathophysiology and treatment of diabetic
peripheral neuropathy: the case for diabetic neurovascular function as an
essential component. Curr. Diabetes Rev. 2006, 2, 131−145.
(4) Rahman, S.; Rahman, T.; Ismail, A. A.; Rashid, A. R. Diabetes-
associated macrovasculopathy: pathophysiology and pathogenesis.
Diabetes Obes. Metab. 2007, 9, 767−780.
(5) Iynedjian, P. B. Molecular physiology of mammalian glucokinase.
Cell. Mol. Life Sci. 2008, 66, 27−42.
(6) van Schaftingen, E. A protein from rat liver confers to glucokinase
the property of being antagonistically regulated by fructose 6-phosphate
and fructose 1-phosphate. Eur. J. Biochem. 1989, 179, 179−184.
(7) Grimsby, J.; Sarabu, R.; Corbett, W. L.; Haynes, N. E.; Bizzarro, F.
T.; Coffey, J. W.; Guertin, K. R.; Hilliard, D. W.; Kester, R. F.; Mahaney,
P. E.; Marcus, L.; Qi, L.; Spence, C. L.; Tengi, J.; Magnuson, M. A.; Chu,
C. A.; Dvorozniak, M. T.; Matschinsky, F. M.; Grippo, J. F. Allosteric
activators of glucokinase: potential role in diabetes therapy. Science
2003, 310, 370−373.
(8) Matschinsky, F. Assessing the potential of glucokinase activators in
diabetes therapy. Nat. Rev. Drug Discovery 2009, 8, 399−416.
(9) Sarabu, R.; Berthel, S. J.; Kester, R. F.; Tilley, J. W. Novel
glucokinase activators: a patent review (2008−2010). Expert Opin. Ther.
Patents 2011, 21, 13−31.
Figure 5. Normoglycemic glucose lowering test of 26 (AM-9514) in
C57Bl/6 mice. Compound 26 showed no hypoglycemia during the test.
Statistical significance compared to vehicle treatment is denoted by
*(p < 0.05), **(p < 0.01), ***(p < 0.001), and ****(p < 0.0001), as
determined by two-way ANOVA, and is color-coded to the treatment in
the figure legends.
(10) Castro, A. Kinase activators as a novel class of antidiabetic agents.
Drug Discovery Today 2012, 17, 528−529.
(11) Pfefferkorn, J. A. Strategies for the design of hepatoselective
glucokinase activators to treat type 2 diabetes. Expert Opin. Drug
Discovery 2013, 8, 319−330.
E
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ACS Medicinal Chemistry Letters
Letter
(12) Hinklin, R. J.; Boyd, S. A.; Chicarelli, M. J.; Condroski, K. R.;
Dewolf, W. E., Jr.; Lee, P. A.; Lee, W.; Singh, A.; Thomas, L.; Voegtli, W.
C.; Williams, L.; Aicher, T. D. Identification of a new class of glucokinase
activators through structure-based design. J. Med. Chem. 2013, 56,
7669−7678.
(13) Lu, M.; Li, P.; Bandyopadhyay, G.; Lagakos, W.; Dewolf, W. E., Jr.;
Alford, T.; Chicarelli, M. J.; Williams, L.; Anderson, D. A.; Baer, B. R.;
McVean, M.; Conn, M.; Veniant, M. M.; Coward, P. Characterization of
a novel glucokinase activator in rat and mouse models. PloS One 2012, 9,
e88431.
(14) For detailed transition from the thiazolylamino pyridine series to
methyl urea-substituted pyridine series, please see: Hinklin, R. J.; Aicher,
T. D.; Anderson, D. A.; Baer, B. R.; Boyd, S. A.; Condroski, K. R.;
DeWolf, W. E., Jr.; Kraser, C. F.; McVean, M.; Rhodes, S. P.; Sturgis, H.
L.; Williams, L.; Houze, J. B. Discovery of 2-pyridylureas as glucokinase
activators. J. Med. Chem. 2014, 57, 8180−8186.
(15) Bonadonna, R. C.; Heise, T.; Arbet-Engels, C.; Kapitza, C.;
Avogaro, A.; Grimsby, J.; Zhi, J.; Grippo, J. F.; Balena, R. Piragliatin
(RO4389620), a novel glucokinase activator, lowers plasma glucose
both in the postabsorptive state and after a glucose challenge in patients
with type 2 diabetes mellitus: a mechanistic study. J. Clin. Endocrinol.
Metab. 2010, 95, 5028−5036.
(16) Meininger, G. E.; Scott, R.; Alba, M.; Shentu, Y.; Luo, E.; Amin,
H.; Davies, M. J.; Kaufman, K. D.; Goldstein, B. J. Effects of MK-0941, a
novel glucokinase activator, on glycemic control in insulin-treated
patients with type 2 diabetes. Diabetes Care 2011, 34, 2560−2566.
(17) Bue-Valleskey, J. M.; Schneck, K. B.; Sinha, V. P.;
Wonddmagegnehu, E. T.; Kapitza, C.; Miller, J. WLY2599506, a novel
glucokinase activator (GKA), improves fasting and postprandial glucose
in patients with type 2 diabetes mellitus (T2DM). Presented at the 71st
American Diabetes Association Meeting, San Diego, CA, 2011.
(18) Pfefferkorn, J. A.; Guzman-Perez, A.; Oates, P. J.; Litchfield, J.;
Aspnes, G.; Basak, A.; Benbow, J.; Bian, J.; Choi, C.; Freeman-Cook, K.;
Corbett, J. W.; Didiuk, M.; et al. Designing glucokinase activators with
reduced hypoglycemia risk: discovery of N,N-dimethyl-5-(2-methyl-6-
((5-methylpyrazin-2-yl)-carbamoyl)benzofuran-4-yloxy)pyrimidine-2-
carboxamide as a clinical candidate for the treatment of type 2 diabetes
mellitus. Med. Chem. Commun. 2011, 2, 828−839.
(19) The S0.5 and Vmax data of Piragliatin and MK-0941 were generated
in house.
(20) Filipski, K. J.; Stevens, B. D.; Pfefferkorn, J. A. New Therapeutic
Strategies for Type 2 Diabetes; RSC Publishing: London, U.K., 2012; pp
88−108.
(21) Grimsby, J.; Berthel, S. J.; Sarabu, R. Glucokinase activators fo rthe
potential treatment of type 2 diabetes. Curr. Top. Med. Chem. 2008, 8,
1524−1532.
(22) Hepatic stability is the ratio of the observed clearance expressed in
mL/min/kg divided by the literature value for the hepatic blood flow
values in the species studied (mL/min/kg).
(23) To save some cost, we used normal C57Bl/6 mice in the OGTT
test of compound 12 instead of ob/ob mice.
(24) Veber, D. F.; Johnson, S. R.; Cheng, Y.-H.; Smith, B. R.; Ward, K.
W.; Kopple, K. D. Molecular properties that influence the oral
bioavailability of drug candidates. J. Med. Chem. 2002, 45, 2615−2623.
(25) Pajouhesh, H.; Lenz, G. R. Medicinal chemical properties of
successful central nervous system drugs. NeuroRx 2005, 2, 541−553.
(26) Please see Supporting Information for detailed experimental
procedure to make 1-(5-bromo-3-((1-ethyl-1H-pyrazol-5-yl)oxy)pyr-
idin-2-yl)-3-methylurea (intermediate 11.3, page 22 in Supporting
Information).
F
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