developing novel biologics for the treatment of diabetes · developing novel biologics for the...
TRANSCRIPT
Developing Novel Biologics for the Treatment of Diabetes
Patricia McDonald PhD
Diabetes Coalition, Palm Beach County
Symposium
The Scripps Research Institute
Jupiter Florida
April 22nd 2016
Financial Disclosure Statement : “I do not have any financial relationships relative to the content of this program.”
Diabetes Coalition, Palm Beach CountySymposium
The Scripps Research InstituteJupiter FloridaApril 22nd 2016
Patricia McDonald PhD
BasicResearch
• Academic Departments
• Metabolism and aging
• Cancer Biology
• Infectious Diseases
• Molecular Therapeutics
• Neuroscience
• Chemistry
Technology Platforms
Cell Based Screening
RNA
Genomics
Proteomics
Drug Discovery
Medicinal Chemistry
uHTS/NIH screening
Discovery Biology
DMPK
In vivo Pharmacology
Structural Biology
Informatics
Translational Research Institute
Scripps Florida
AdvancedTechnologies
DrugDiscovery
Drug Development Process
Discovery Development
3-5 Years +7 Years +1.5 Years
‘Bench to Bedside’
Basic ResearchDone at the Bench
Clinical ResearchDone in the Patient
Translational ResearchBench -to- Bedside
Therapeutic Target Classes
GPCRs 45%
Unknown 7%
Ion channels 5%
Nuclear receptors 2%
Enzymes 28%
Hormones and factors 11%
Nucleic acids 2%
The current therapeutic classes in the pharmaceutical portfolio can be subdivided
into seven main classes, wherein GPCRs represent the largest class.
Selected Examples of Top-Selling
Drugs Targeting GPCRs
GPCR target Drug Disease Company 2000 sales(US $m)
Histamine Receptors Zantac Ulcers GSK 870
Pepcid Ulcers Merck 850
Claritin Allergies Schering Plough 2200
Allegra Allergies Aventis 1100
Serotonin Receptors Risperdal Psychosis Johnson & Johnson 1600
Imitrex Migraine GSK 1100
BuSpar Anxiety Bristol-Myers Squibb 714
Zyprexa Schizophrenia Eli Lilly 3200
Angiotensin Receptors Cozaar Hypertension Merck 1700
Toprol-XL Hypertension AstraZeneca 580
Adrenoreceptors Coreg CHF GSK 250
Serevent Asthma GSK 940
Muscarinic Acetylcholine Atrovent COPD Boehringer Ingelheim 600
GnRH Receptors Zoladex Cancer AstraZeneca 740
Dopamine Receptors Requip Parkinson's GSK 90
Prostaglandin Receptors Cytotec Ulcers Pharmacia 100
ADP Receptors Plavix Stroke Bristol-Myers Squibb 900
GABAB Receptor Gabapentin Pain 1334
~40% of all modern drugs and ~25% of the top 100 best selling drugs target GPCRs
C
1 2 3 4 5 6 7
N
Extracellular
Intracellular
G Protein-Coupled Receptors: GPCRs
ActivatorsLight, odorants
HormonesNeurotransmitters
PeptidesBiogenic amines
Biological FunctionsSensory perceptionNeurotransmissionEndocrine functionExocrine function
ChemotaxisControl of blood pressure
EmbryogenesisDevelopmentCell growth
Cell differentiationHIV infection
ai as aq a12
Ion channels
Inhibition cAMP
Phopholipases
Phophodiesterases
Inhibition cAMP PLC-bDAG
Ca2+
PKC
Rho GEFs
Rho
Ga bg
bg
PLC-bcAMP
Ion channels
Glucagon Like Peptide -1 Receptor: GLP-1R
Peripheral and Central Actions of GLP-1R and GLP-1
GLP-1R Activation and Signaling
• Compromised b cell function
• Pancreatitis• Pancreatic cancer• Thyroid cancer
?
• Nausea• Vomiting• Diarrhea/Constipation• Abdominal Pain
• Enhances glucose-dependent
insulin secretion
• Reduces hepatic glucose output
• Regulates gastric emptying
• Promotes satiety and
reduction of appetite
• Increase in β-cell mass?
GLP-1R
GLP-1
Novel Approach: Signal Selectivity
Current Industry Approach: Target Specificity
Compound & Assay
Plate Storage
Stäubli RX130L robotic arm
Plate Reader
Compound Transfer Station
High Throughput Screening (HTS)Automation Platform
Cell Culture Suite
Development of an Autocrine-Based System forScreening Large Combinatorial Peptide Libraries
~100 million
peptides/month
GLP-1R
P5: The First Potent, Selective GLP-1R G-protein Biased Agonist
Gasb-arrestin
GLP-1R
Peptide
1,2,3….
P5 Improves Glucose Tolerance but Displays Weak Insulin Secretagogue Activity
C57BL/6-DIOAcute GTT 8 hr Fasting
0 1 5 3 0 4 5 6 0 7 5 9 0
1 0 0
2 0 0
3 0 0
4 0 0
5 0 0
6 0 0
7 0 0
T im e (m in )
Glu
co
se
(m
g/d
l)
C o n t r o l
E x 4 (1 0 u g /k g )
P 5 (1 0 u g /k g )
**
**
**
**
**
**
**
******
0 1 5 3 0 4 5 6 0 7 5 9 0
1 0 0
2 0 0
3 0 0
4 0 0
5 0 0
6 0 0
7 0 0
T im e (m in )
Glu
co
se
(m
g/d
l)
c o n t r o l
E x 4 (1 u g /k g )
P 5 (1 u g /k g )
**
***
**
co
ntr
ol
P5 (
10 u
g/k
g)
P5 (
1 u
g/k
g)
Ex4 (
10 u
g/k
g)
Ex4 (
1 u
g/k
g)
-0 .6
-0 .4
-0 .2
0 .0
0 .2
g
lyc
ate
d H
bA
1c (
%)
*
co
ntr
ol
P5 (
10 u
g/k
g)
P5 (
1 u
g/k
g)
Ex4 (
10 u
g/k
g)
Ex4 (
1 u
g/k
g)
0
1 0 0
2 0 0
3 0 0
4 0 0
5 0 0
Glu
co
se
(m
g/d
l)
** ***
P5 has Superior Antihyperglycaemic Efficacy in DIO Mice
GTT 8 hr Fasting in following 4wk treatment in DIO
Non-fasting
Chronic Study
HbA1c
-4 -3 -2 -1 0 1 2 3 4 5
0
2 0
4 0
6 0
8 0
1 0 0
1 2 0
lo g [p e p tid e ] n M
Ce
llu
lar R
es
po
ns
e (
% o
f m
ax
)
P 5
E x 4
P 5 -F c
EC50=5 nM
In Vitro Characterization of Long-acting G-protein Biased Agonist P5-Fc
P5
a b
c
NH2 NH2
COOHCOOH
0 1 5 3 0 4 5 6 0 7 5 9 0
0
5 0
1 0 0
1 5 0
2 0 0
2 5 0
3 0 0
3 5 0
4 0 0
4 5 0
5 0 0
5 5 0
T im e (m in )
Glu
co
se
(m
g/d
l)
c o n tro l
P 5 -F c
P 5
0 1 5 3 0 4 5 6 0 7 5 9 0
0
5 0
1 0 0
1 5 0
2 0 0
2 5 0
3 0 0
3 5 0
4 0 0
4 5 0
5 0 0
5 5 0
T im e (m in )
Glu
co
se
(m
g/d
l)
c o n tro l
P 5 -F c
P 5
0 1 5 3 0 4 5 6 0 7 5 9 0
0
5 0
1 0 0
1 5 0
2 0 0
2 5 0
3 0 0
3 5 0
4 0 0
4 5 0
5 0 0
5 5 0
T im e (m in )
Glu
co
se
(m
g/d
l)
c o n tro l
P 5 -F c
P 5Day 0 Day 2 Day 7
P5-Fc Displays Enhanced Pharmacodynamics
Time-dependent blood glucose lowering activity of P5-Fc
fusion protein in lean mice. (a) Glucose tolerance tests
performed either directly following a single injection of P5
(20nmol/kg) or P5-fc (5 nmol/kg) (day 0) or two and seven
days later. (b) Effect of a single injection of P5-fc (50 nmol/kg)
on fed blood glucose level.
0 2 4 6 8 1 0 1 2 1 4
0
1 0 0
2 0 0
3 0 0
4 0 0
D a y s
Glu
co
se
(m
g/d
l)
P 5 -F c (5 0 n m o l/k g )
c o n t ro l
**** ** ** ** ** ** ** **
**
**
** ** **
****
** **
a
b
Non-fasting DIO
GTT Fasting Lean Mice
0 1 5 3 0 4 5 6 0 7 5 9 0 1 0 5 1 2 0
0
1 0 0
2 0 0
3 0 0
4 0 0
5 0 0
T im e (m in )
Glu
co
se
(m
g/d
l)
Q W P 5 -F c (1 0 n m o l/k g )
C o n t ro l
Q W P 5 (5 0 n m o l/k g )
**
**
**** **
P5-Fc Displays Antihyperglycaemic Efficacy in DIO Mice(one week after injection 5)
co
ntr
ol
P5-F
c (
QW
10 n
mo
l/kg
)
P5-F
c (
QW
50 n
mo
l/kg
)
0
5 0
1 0 0
1 5 0
2 0 0
2 5 0
Glu
co
se
(m
g/d
l)
** **
Glucose tolerance test. (a) Effect of once-weekly administration of P5-Fc on glucose tolerance in
response to glucose challenge (one week after injection 5 and before injection 6). (b) Effect of once-
weekly administration of P5-Fc on fed plasma glucose levels (one week after injection 5 and before
injection 6) and (c) on HbA1c (after injection 5). n=5; *, p<0.05; **, p<0.01.
a b c
co
ntr
ol
P5-F
c (
QW
10 n
mo
l/kg
)
P5-F
c (
QW
50 n
mo
l/kg
)
4 .0
4 .5
5 .0
5 .5
Hb
A1
c (
%)
*
P=0.015
Non-fastingGTT-Fasting HbA1c
Chronic Study
0 1 5 3 0 4 5 6 0 7 5 9 0 1 0 51 2 0
0
5 0
1 0 0
1 5 0
2 0 0
2 5 0
T im e (m in )
Glu
co
se
(m
g/d
l)
Ins + 10 nm ol/kg P5-Fc
Ins
Ins + 50 nm ol/kg P5-Fc
P5-Fc Enhances Insulin Sensitivity in DIO Mice
**
**
** **
**
Insulin tolerance test. (a) Effect of once-weekly administration of P5-Fc on glucose level in response to
insulin injection (0.75 U/kg), ITT performed one week after injection 4 and prior to injection 5).
(b) Data are represented as a % of basal blood glucose level for each individual group. n=5; *, p<0.05;
**, p<0.01.
A)
**
**** **
****
**
**
**
ITTB)
Ins (
0.7
5U
/kg
)
In +
QW
P5-F
c (
10 n
mo
l/kg
)
Ins +
QW
P5-F
c (
50 n
mo
l/kg
)
0
5 0 0 0
1 0 0 0 0
1 5 0 0 0
Ins
uli
n A
UC
d a y s o f tre a tm e n t
cu
mu
lati
ve
fo
od
in
tak
e (
g)
-2 0 2 4 6 8 1 0 1 2 1 4 1 6 1 8
0
1 0
2 0
3 0
4 0
5 0
P 5 -F c (Q W ,1 0 n m o l/k g )
C o n t ro l
P 5 -F c (Q W ,5 0 n m o l/k g )
P5-Fc Decreases Food intake and Body Weight in DIO Mice
**
Effect of once weekly administration of P5-Fc on cumulative (A) and daily food intake (B).
P5-Fc significantly decreases food intake for 2-3 days post injection. n=5; *, p<0.05; **, p<0.01.
A)
Injection
Injection
Injection
-2 0 2 4 6 8 1 0 1 2 1 4 1 6 1 8
-6
-5
-4
-3
-2
-1
0
1
2
c o n t ro l
P 5 -F c (Q W ,1 0 n m o l/k g )
P 5 -F c (Q W ,5 0 n m o l/k g )
d a y s o f tre a tm e n t
W
eig
ht
(g)
Injection
Injection Injection
B)
• Using a high throughput autocrine-based approach we enabled the functional
screening of large combinatorial peptide libraries with intrinsically high diversity.
• We have identified a novel, highly potent, selective G-protein biased agonist of the
GLP-1R, designated P5.
• P5 demonstrates decreased insulinotropic activity but potent antidiabetic properties.
• P5 improves insulin sensitivity, long term glycemic control and has superior effects on
HbA1c levels compared to Exendin 4, suggesting that GLP-1R G-protein-biased
agonists may provide a novel therapeutic approach to T2DM.
• P5-Fc promotes weight loss.
• P5-Fc has enhanced PD compared to P5 allowing once weekly treatment.
Summary
Discovery Development
3-5 Years +7 Years +1.5 Years
P5
McDonald lab
Emmanuel Sturchler
Ainhoa Nieto
Rachel Turn
Richard Hawkins
*
Lerner Lab
Hongkai Zhang
Jia Xie
Teresa Jones
LinLing He
AcknowledgementsDawson Lab
Philip Cistrone