The human approach to target The human approach to target validation and drug discoveryvalidation and drug discovery

Dr. Alastair J. RiddellDr. Alastair J. RiddellChief Executive OfficerChief Executive OfficerPharmagenePharmagene

33rdrd October, Tokyo October, Tokyo

Presentation outline

Introduction to Pharmagene

The Integration of Pharmagene’s products and services— TargetEvaluator™, Phase ZERO ™, Indication Switch ™— Into the new Custom Validation Programme

Pharmagene Therapeutics— Cystic fibrosis— Irritable bowel syndrome— Migraine

Pharmagene’s human tissue resource

Established relationships with suppliers in UK, NL and US

clinical history with each sample no reliance on single supplier of tissue

types

Dynamic and expanding resource — > 500,000 samples from >5,000 donors— 80% living, 20% autopsy

Legal & ethical integrity— tissue accepted only after informed consent— donor anonymisation

No supply to third parties

Biomaterials received and processed 24hours a day, 7 days a week using SOP’s

Managed by in-house Consultant Pathologist

0060504L 4

Global customer base

US

Bayer (CT&CA) AllerganCubistJ&JBristol-Myers Squibb

Europe

GlaxoSmithKline Merck Sharp & DohmePfizerAstraZenecaRocheBoehringer-IngelheimBayer (UK)Janssen (J&J)Amersham FerringOxford BioMedicaVernalis

Japan

Kyowa Hakko KogyoSankyoBayer YakuhinDaiichiTaishoT*Ono

Compound Compound ValidationValidation

PharmageneCustom Validation Platform

PharmageneIndication Switch ™

PharmageneTargetEvaluator™

PharmagenePhase ZERO™

GenomicInformation

CompoundScreening

A solution provider..

Cellular localisation(mRNA and protein)

TargetTargetValidationValidation

PharmagenePharmageneCustom Validation PlatformCustom Validation Platform

ProteinProteinLocalisationLocalisation

Cell-basedCell-basedAssay Assay DevelopmentDevelopment

FunctionalFunctionalAnalysisAnalysis

ExpressionExpressionAnalysisAnalysis

Stage 1 Stage 2 Stage 3

A staged integrated approach..

Stage 4

Pharmagene Validated Drug TargetPharmagene Validated Drug Target

Validated TargetValidated TargetValidated TargetValidated Target

Stage 1Stage 1Expression AnalysisExpression Analysis

Stage 1Stage 1Expression AnalysisExpression Analysis

Stage 2Stage 2Protein LocalisationProtein Localisation

Stage 2Stage 2Protein LocalisationProtein Localisation

Stage 3Stage 3Cell based assay Cell based assay

developmentdevelopment

Stage 3Stage 3Cell based assay Cell based assay

developmentdevelopment

Stage 4Stage 4Functional AnalysisFunctional Analysis

Stage 4Stage 4Functional AnalysisFunctional Analysis

Directed access toDirected access to

TargetEvaluatorTargetEvaluatorDirected access toDirected access to

TargetEvaluatorTargetEvaluator

Antibody localisation studiesAntibody localisation studies

Target analysis in specifiedTarget analysis in specifiedtissues and Pathologist reporttissues and Pathologist report

Antibody localisation studiesAntibody localisation studies

Target analysis in specifiedTarget analysis in specifiedtissues and Pathologist reporttissues and Pathologist report

For Target or ligand :For Target or ligand :

Cell based AssaysCell based Assays

Demonstrate functionalDemonstrate functionalresponse on target activationresponse on target activation

For Target or ligand :For Target or ligand :

Cell based AssaysCell based Assays

Demonstrate functionalDemonstrate functionalresponse on target activationresponse on target activation Ligand studies in targetLigand studies in target

tissuestissues

Pharmacology in peripheral Pharmacology in peripheral

and CNS tissueand CNS tissue

Ligand studies in targetLigand studies in target

tissuestissues

Pharmacology in peripheral Pharmacology in peripheral

and CNS tissueand CNS tissue

TargetsTargets

Custom Validation Platform - overview

Custom Validation Platform

Human tissue based

Custom design to allow validation of targets, compounds or both

Multiple stages with the ability to stop at any stage

Regular discussion between both parties

Flexible deal structure, based on success

Pharmagene TherapeuticsPharmagene TherapeuticsIn-house programmesIn-house programmes

Pharmagene Therapeutics

Current research areas– Respiratory – Gastrointestinal – Cardiovascular– Inflammatory diseases

— Multiple targets identified and undergoing validation— Intellectual property portfolio expanded in all areas

Three lead programmes– Cystic Fibrosis (PGN0052) – Irritable Bowel Syndrome (R1)– Migraine (R4)

— All with novel mechanisms of action

Tertiary Bronchus

0

2000

4000

6000

8000

10000

CF (n = 24)

Control (n = 17)

Co

py

nu

mb

er *

4x

3ry bronchus & lung

parenchyma

R52 ligand as a treatment for CF

Significantly up-regulated in bronchus from CF patients

R52 riboprobe in-situ hybridisation in tertiary bronchus

sense antisense

donor 1

donor 2

donor 3

R52 Ab Immunocytochemistry in CF Tertiary Bronchus

x200 mag

100g/ml

+ 100x blocking peptide

IgG control

R52 and ion flux in bronchus

Time (min)

Isc (2µA)

0 5 10 15 20 25 30 35

amiloride 10 M (apical)

R52 ligand 3 M (apical)

ATP 10 M (apical)

Cystic Fibrosis – PGN0052, an Indication Switch

Cystic Fibrosis is a monogenic disease attributed to a defect in a chloride transport mechanism

— Fatal disease with few effective pharmacological treatments

R52 is a well known hormone receptor for a hormone that stimulates chloride transport

TargetEvaluatorTM revealed the novel discovery of R52 expression in the lung

TaqMan RT-PCR studies showed R52 up-regulated 4-fold in tertiary bronchus of CF

Phase ZERO technologies demonstrated that PGN0052 stimulates chloride transport in tertiary bronchus of the lung

PGN0052 programme is being progressed to enter the clinic in Q4 2002

Irritable Bowel Syndrome - R1- New Lead Discovery from Indication Switch

Significant unmet medical need and commercial opportunity

5-Hydroxytryptamine (5-HT) has long been implicated in IBS

— Large investment by Pharma in 5HT3 antagonists & 5HT4 agonists (Lotronex & Zelmac)

— Current drugs have not delivered success

0060504L.ppt 18

GI tract

5-HT2B antagonists as a treatment for IBS

Borman et al. Br J Pharmacol (in press)

5-HT2B receptor protein expression

Protein dot blots with 5-HT2B Ab

Tissue Whole MucosaSmoothmuscle

TaeniaColi

Ileum NA

Colon

• Protein localised in colon smooth muscle

Borman et al. Br J Pharmacol (in press)

Irritable Bowel Syndrome - R1

Pharmagene Target EvaluatorTM revealed significant expression of 5HT2B receptor in human colon

Immunocytochemical stains revealed the highest concentration of the receptor in nerve plexi and longitudinal smooth muscle

C

L

NP

Functional evidence for 5-HT2B receptors in colon

Smooth muscle pharmacology

• Effect blocked by the potent, selective 5-HT2B antagonist, RS-127445

RS-127445 100nM

Electrically-induced contractions of colonic strips

• In human colon, 5-HT causes smooth muscle hypersensitivity, and this effect is mediated by 5-HT2B receptors

100-fold

Borman et al. Br J Pharmacol (in press)

Irritable Bowel Syndrome - R1

Pharmagene Target EvaluatorTM revealed significant expression of 5HT2B receptor in human colon

Phase ZERO technologies established a novel role for the 5HT2B receptor

— Enhances the contractile response of human colon to nerve stimulation

Selective antagonists at 5HT2B receptor represent a completely novel mechanism of action for potential treatment of IBS (example RS 127445)

Pharmagene has identified two entirely novel lead series with enhanced performance over RS 127445. Early stage lead compounds identified.

Optimised leads ready for licensing H1 2003

Migraine - R4 - New Lead Discovery

Significant advances in the treatment of migraine

However, side-effects of current therapies mean continuing unmet medical need and commercial opportunity

B

PP

M

A

40mm

Human cerebral arterial vascular tree Measuring the function of human cerebral artery in vitro

Migraine - R4 - New Lead Discovery

Pain of migraine associated with dilation of the cerebral arterial vasculature

Circulating levels of PGE2 reported to be increased in migraine

Detailed pharmacological studies of the human cerebral artery in vitro revealed dilator effect of PGE2. This response is mediated by the EP4 receptor

Pharmagene has a granted UK patent for the use of EP4 antagonists to treat primary headache disorders

Collaboration with Argenta to identify selective EP4 antagonists through an intelligent screening approach

Optimised leads ready for licensing by end 2003

Pharmagene Drug Discovery

Based on an integrated human approach

Validated by global customer base and growing revenues

Most customers are repeat and long term

The integrated technologies have produced novel in-house therapeutic opportunities

Lead candidates ready for licensing over next 1-2 years in — Irritable Bowel syndrome— Migraine— Cystic Fibrosis