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Full scale of diabetes assays and models
To accelerate your drug discovery process
www.wuxiapptec.com
Enabling Unbounded Possibilities
2000 2010 20300
100
200
300
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500
151
285
438
No
. o
f D
iab
ete
s (
Millio
n)
• 6.4% of adult population in 2010
• 54% increase 2010 to 2030
• 36% in India and China alone
China: 2007-2008
(NEJM 362;12, 2010)
China: 2010
(JAMA. 2013, 310:948)
9.7% Diabetes
15.5% Prediabetes
11.6% Diabetes
50.1% Prediabetes
AMPK, PPARragonist
Blood glucose
5.6 mM/100 mg/dl
3.9 mM/70 mg/dl
Normal range
Insulin
Insulin analog
GLP-1 , GIP
&Aanalog
DPPV inhibitor
SGLT2 inhibitor
Hepatic Glucose
output Glucose uptake and
utilization
Glucose
reabsorption
GLP-1 agonist
KATP channel
inhibitor
AMPK agonist
GCGR
antagonist
Glucose absorption
-glucosidase
inhibitorHyperglycemia
Reference: DRCP, 2010 , 87:4
New drug R&D
In vitro pharmacological
profiling of compounds,
insulin and/or biosimilars
Biosimilar equivalence
assessment, quality testing
IND filling
• Complete recording and
achieving of samples, assay
procedures, raw data and
results
• Data authenticity
• Assay traceability
• CFDA on site inspection1
Category
Name/Target
(human if not
specified)
Cell lineLigand binding
/Enzyme assayFunction assay
INS & Analog
Insulin-R A (short) CHO I-125 phosphorylation
Insulin-R B (long,
exon 11)CHO I-125 phosphorylation
IGF-1R H19-7 I-125 phosphorylation
IR/GLUT4
3T3-L1
adipocytes
[3H]Deoxy-D-
glucoseglucose uptake
IR [14C]-glucose lypogenesis
IR [14C]-glucose Glycogen formation
GLP1R GLP-1R HEK293 FRET/I-125 cAMP
GCGR Glucagon R HEK293 I-125 cAMP, Ca
SGLT
SGLT1 CHO[14C]Methyl a-D-
glucopyranoside
(AMG)SGLT2 CHO
SGLT2 (rat) CHO
DPP DPP4，8， 9 Luminescent
NHR PPAR-gamma Fluorescent
2
Affinity: Binding Assay
Method: Radioligand filtration binding assay
Format: 96-well
Insulin receptor-A binding assay
-2 -1 0 1 2 3 4
0
20
40
60
80
100
120
Insulin
IC50 = 1.4 nM
Log Con. [nM]
% In
hib
ito
n
Insulin receptor-B binding assay
-3 -2 -1 0 1 2 3
0
20
40
60
80
100
120
Insulin
IC50 = 1.2 nM
Log Con. [nM]
% In
hib
ito
n
Stable cell line: CHO-INSRA, WuXi Stable cell line: CHO-INSRB, WuXi
Function: Phosphorylation Assay
Method: Alpha screen assay (phosphorylation)
Format: 384-well
Insulin Receptor-A function assay
-2 -1 0 1 2 3 4
0
20
40
60
80
100
120
Insulin
EC50 = 10.6 nM
Log Con. [nM]
% A
cti
vit
y
Insulin Receptor-B function assay
-2 -1 0 1 2 3 4
0
20
40
60
80
100
120
Insulin
EC50 = 15.1nM
Log Con. [nM]
% A
cti
vit
y
Stable cell line: CHO-INSRA, WuXi Stable cell line: CHO-INSRB, WuXi
Glucose Uptake Assay
Cell line: 3T3-L1, ATCC
Method: Radioactive assay
Format: 96-well
Glucose uptake in 3T3-L1 adipocytes
-3 -2 -1 0 1 2 3-20
0
20
40
60
80
100
120Insulin
EC50 = 2.0 nM
Log Con. [nM]
% A
cti
vit
y
Adipocytes
Induction
3T3-L1 (ATCC, CL-173™)
Cell line: 3t3-L1, ATCC
Method: Radioactive assay
Format: 96-well
Lipogenesis in 3T3-L1 adipocytes
-4 -2 0 2 4-50
0
50
100
150Insulin
EC50 = 0.71 nM
Log[Con.nM]
Acti
vit
y%
Glycogen synthesis in 3T3-L1 adipocytes
-4 -2 0 2 4-50
0
50
100
150Insulin
EC50 = 1.42 nM
Log[Con.nM]
Acti
vit
y%
Cell line: 3t3-L1, ATCC
Method: Radioactive assay
Format: 96-well
Function: cAMP Assay
Method: TR-FRET cAMP assay
Format: 384-well
-16 -14 -12 -10 -8 -6 -40
2
4
6Exenatide
GLP-1 receptor binding assay (Tag-lite)
IC50 = 6.2 nM
Log Con. [nM]
HT
RF
Rati
o (
665/6
20)
GLP-1 receptor cAMP assay
-6 -4 -2 0 2 4-20
0
20
40
60
80
100
120
140
Exenatide
EC50 = 0.06 nM
Log Con. [nM]
% A
cti
vit
y
Affinity: Binding Assay
Method: Tag-lite assay
Format: 384-well
Stable cell line: HEK-GLP-1R, WuXi Stable cell line: HEK-GLP-1R, WuXi
Affinity: Binding Assay
Method: Radioligand filtration binding assay
Format: 96-well
Function: cAMP Assay
Method: TR-FRET cAMP assay
Format: 384-well
GCGR receptor binding assay
-2 -1 0 1 2 3 4
20
40
60
80
100
120
Glucagon
IC50 = 7.7 nM
Log Con. [nM]
% In
hib
ito
n
GCGR receptor cAMP assay-antagonist mode
-2 0 2 4 6-20
0
20
40
60
80
100
120
MK-0893
IC50 = 37.7 nM
Log Con. [nM]
% In
hib
ito
n
Stable cell line: HEK-GCGR, WuXi Stable cell line: HEK-GCGR, WuXi
Primary Screening
Enzyme: DPP4
Method: Luminescent assay
Format: 384-well
DPP4 inhibition assay
-14 -12 -10 -8 -6
0
20
40
60
80
100
120
Linagliptin
IC50 = 0.29 nM
Log Con. [M]
% C
on
tro
l
PPAR-gamma Binding Assay
0 1 2 3 4 50
25
50
75
100
125
150
175Troglitazone
IC50 = 408 nM
Log Con. [nM]
Po
lari
zati
on
(m
P)
3
Primary ScreeningEnzyme: PPARγ-LBDMethod: Fluorescence polarization
Format: 384-well
Transporter assay
Method: Radioactive assay
Format: 96-well
hSGLT1 transporter assay
-2 0 2 4 6-20
0
20
40
60
80
100
120
Dapagliflozin
IC50 = 417.5 nM
Log Con. [nM]
% In
hib
itio
n
hSGLT2 transporter assay
-4 -2 0 2 4-20
0
20
40
60
80
100
120
Dapagliflozin
IC50 = 0.74 nM
Log Con. [nM]
% In
hib
itio
n
Stable cell line: CHO-hSGLT2, WuXi Stable cell line: CHO-hSGLT1, WuXi
Fat NHP:
High fat diet induced, spontaneous
Diabetes study:
• Oral Glucose Tolerance Test (OGTT)
• Mixed Meal Tolerance Test (MMTT)
• Intravenous Glucose Test (IVGTT)
• Insulin Tolerance Test (ITT)
• Graded Glucose Infusion (GGI)
• Glucose Clamping
Glucose Clamping
20% Glucose
solution
Continuous i.v.
infusion Serial blood sampling
Rapid plasma
glucose measurement
Glucose infusion rate adjustment to achieve plasma steady state
Insulin
solution
4
Imaging:
• MRI liver fat content quantitation
• MRI liver fibrosis detection
• MRI blood vessel imaging
• Body composition measurement
by DEXA
Tissue biopsy:
• Liver
• Kidney
• Muscle
• Fat
• Brain
Graded Glucose Infusion
Introduction:
An assay to assess glucose tolerance including oral glucose
(OGTT), intraperitoneal glucose(IPGTT) or intravenous glucose
(IVGTT) to fasted animals
Species: Mouse, rat and NHP
Endpoint: Blood glucose and Insulin
Throughput: 8 animals/group; 5~8 groups
Groups in a standard assay: Vehicle, Positive control, Testing
compounds with 3 doses
Turnaround time : ~5 days/each study
Effect of Cpd A (s.c)on Blood Glucose lowering
time course in IPGTT
0
10
20
30
40
0 30 120-60
Glucose
20 40
Test article
60
Cpd A High
VehicleCpd A LowCpd A Medium
Exenatide
Time(min)
Blo
od
glu
co
se(m
mo
L/L
)
Effect of Cpd A (s.c) on reducing Glucose AUC 0-120min in IPGTT
Data were presented as Mean ± SEM, n=5 per group, ***p<0.001
vs. vehicle, one way ANOVA followed by Dunnett’s test
Introduction:
An assay to measure urine glucose
Species: Rat and NHP
Endpoint: Urine volume; Urine glucose; Blood glucose
Groups in a standard assay: Naïve(Vehicle), Positive
control, Testing compounds with 3 doses; n=4~6
animals/group
Turnaround time : 2 days/each study
0
1 0
2 0
3 0
4 0
Urin
e v
olu
me
(m
L/2
00
g B
W) ***
E ffe c t o f C a n a g lif lo z in (p .o .) o n u r in e v o lu m e
in 2 4 h rs p o s t tre a tm e n t
C a n a g liflo z in 1 m p k
V e h ic le
C a n a g liflo z in 3 m p k
C a n a g liflo z in 1 0 m p k
0
1 0 0 0
2 0 0 0
3 0 0 0
4 0 0 0
5 0 0 0
Urin
e g
luc
os
e (
mg
/20
0 g
BW
)
***
***
***
E ffe c t o f C a n a g lif lo z in (p .o .) o n th e u r in e g lu c o s e e x c lu s io n
in 2 4 h rs p o s t t re a tm e n t
C a n a g lif lo z in 3 m p k
C a n a g lif lo z in 1 0 m p k
C a n a g lif lo z in 1 m p k
V e h ic le
V eh ic le ≈0
Data were presented as Mean±SEM, n=5 per group, *** p＜0.001 vs vehicle
group, one way ANOVA followed by Dunnett multiple comparison test.
5
Introduction:
A commonly used model to assess the anti-diabetic
effect in db/db mice
Species: Mouse
Model: C 57 BL/KsJ.db male mice
Endpoint:
Body weight, food intake, water intake, blood
glucose, GTT, insulin, GLP-1, GTT,
HBA1C/fructosamine, pancreases histology stain
Throughput: 10~12 animals/group; 5~8 groups
Groups in a standard assay: Vehicle, Positive control,
Testing compounds with 3 doses
Turnaround time : ~6 weeks/each study
Effect of Rosiglitazone and DMBG (p.o) on Blood Glucose (mmol/L)in db/db mice
0
5
10
15
20
25
30
35
db/m wild type vehicle
db/db vehicle
db/db Rosiglitazone
db/db DMBG
****** ******
*** *** *** ***
Blo
od
Glu
co
se (m
mm
ol/L
)
Baseline Week1 Week2 Week3
Data were presented as Mean ± SEM, n=12 per group, *p<0.05,**P<0.01,
***p<0.001 vs. vehicle, one way ANOVA followed by Dunnett’s test
7-8 weeks old db/db mice were treated with Cpds for three weeks.
Introduction:
A commonly used model to assess the anti-diabetic
effect in ZDF rat
Species: Rat
Model: Zuker fa/fa male rats
Endpoint:
Body weight, food intake, water intake, blood glucose,
insulin, GLP-1, GTT, HBA1C/fructosamine, pancreases
histology stain
Throughput: 8 animals/group; 5~8 groups
Groups in a standard assay: Vehicle, Positive control,
Testing compounds with 3 doses
Turnaround time : ~6 weeks/each study
Data were presented as Mean ± SEM, n=8 per group, *p<0.05
vs. vehicle, one way ANOVA followed by Dunnett’s test
7-10 weeks old ZDF rats were treated with Rosiglitazone for three weeks.
Effect of Rosiglitazone on Blood Glucose in ZDF rats
Introduction:
• Mice fed with HFD (Research Diet D12492 ) for 12~14
weeks to assess anti-obesity agents
• 200 DIO mice /month ready to use
Species: Mouse and NHP
Endpoint:
Body weight, food intake, GTT, total Cholesterol (TC),
Triglyceride (TG), ALT, AST, TG in, liver/muscle Insulin,
GLP-1, HBA1C ,Leptin, HE/Oil red O stain, Fat pad, etc.
Throughput: 8 mice/group; 5~8 groups
Groups in a standard assay: Vehicle, Positive control,
Testing compounds with 3 doses
Turnaround time : ~6 weeks/each studyAnti-obesity effect of Rimonabant chronic treatment
Data were presented as Mean ± SEM, n=8 per group, *p<0.05, **p<0.01 vs.vehicle, one way ANOVA followed by Dunnett’s test
6
Introduction:
A model induced by feeding with High Fructose Diet (HFD)
to assess dyslipidemia
Species: Rat and NHP
Endpoint: Body weight, food intake, Total cholesterol (TC),
Triglyceride (TG), HDL-C, LDL-C, etc.
Throughput: 8 rats/group; 5~8 groups
Groups in a standard assay: Vehicle, Positive control, Testing
compounds with 3 doses
Turnaround time : ~4 weeks/each study
Data were presented as Mean ± SEM, n=8 per group, *p<0.05,**P<0.01,
***p<0.001 vs. vehicle, one way ANOVA followed by Dunnett’s test
Effect of EPA on Plasma TG levels
Enabling Unbounded Possibilities
Contact
Henry Lu
Vice President, Biology
Changqing Cao
Executive Director
Wei Liang
Director
Longji Xu
Director, Business Development
