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