Translation of Safety Biomarkers in the Clinical Setting

Michael MerzNovartis Institutes for BioMedical Research

Ina Schuppe KoistinenGlobal Safety Assessment, AstraZeneca R&D, Sweden

Coordinators IMI SAFE-T consortium

24th AnnualEuroMeeting

26-28 March 2012Copenhagen, Denmark

DisclaimerThe views and opinions expressed in the following PowerPoint slides are those of the individual presenter and should not be attributed to Drug Information Association, Inc. (“DIA”), its directors, officers, employees, volunteers, members, chapters, councils, Special Interest Area Communities or affiliates, or any organization with which the presenter is employed or affiliated.

These PowerPoint slides are the intellectual property of the individual presenter and are protected under the copyright laws of the United States of America and other countries. Used by permission. All rights reserved. Drug Information Association, DIA and DIA logo are registered trademarks or trademarks of Drug Information Association Inc. All other trademarks are the property of their respective owners.

2

Outline

• IMI SAFE-T Consortium: brief overview

• Status 2012

• Clinical biomarker qualification program

• Focus drug-induced liver injury: biomarkercandidates

• Initial experimental results

• Collaborations

3

The IMI SAFE-T* ConsortiumScope

Three organs needing better clinical monitoring ofdrug-induced injuries:

Kidney: current standards increase only once 50-60% of kidney function is lost.

Liver: current standards are not sufficiently sensitive and specific and do not adequately discriminate adaptors from patients at high risk to develop liver failure.

Vascular System: currently no biomarkers available for drug-induced vascular injury in human.

44

*Safer And Faster Evidence-based Translation

• Patient level – Lower injury threshold

– Earlier time to onset

– Larger extent of changes

– Improved specificity

– Better suited to monitor and predict clinical course

– Better suited to assess causality

• Population level – Earlier and more specific signal detection in clinical development programs

– Improved mechanistic insight

– Superior in terms of identifying underlying pathology

– Better suited to predict human risk from animal toxicity

Biomarker attributes of interest

5

Key challenges for biomarker qualification

• Substantial background variability in initial candidate markers

• Biomarker response varies across different populations

• Large initial number of biomarker candidates requires substantial sample volumes to be taken

• Key target responses, i.e. specific adverse drug reactions, suitableand accessible for qualification are overall very rare

Large sample sizes are required

Multitude of patient populations need to be included

6

Qualification cannot be achieved by one company alone

SAFE-T participantsAcademiaAcademia

CollaboratorsCollaboratorsAdvisorsAdvisors

SMEs

7

IMI SAFE-T ConsortiumObjectives

• To evaluate utility of safety biomarkers for detecting, assessing, and monitoring drug induced kidney, liver, and vascular injury in humans

• To develop assays and devices for clinical application of safetybiomarkers

• To compile enough evidence to qualify safety biomarkers for regulatory decision making in clinical drug development and in a translational context

• To gain evidence for how safety biomarkers may also be used in the diagnosis of diseases and in clinical practice

8

Funding and timing

Financing• IMI funding: 13.9 mio EUR• EFPIA contribution, mainly in kind: 17.7 mio EUR• Contribution academia/SME: 4.1 mio EUR

• Total project cost: 35.7 mio EUR

Timing:• Starting date: June 15, 2009• Duration: Five years

9

DILI BM step 1 list

Submit to health authorities

Literature

SAFE-T sources

DatabasesEvaluation

DILI BM step 2 list

Healthy volunteers

Patients non-liver disease

Patients liver disease

Patients hepatotoxic drugs

Samples

Regulatory advice

DILI BM f inal list

DILI BM step 3 list

Assay / stat analysis / select BMs

Regulatory advice

Regulatory approval

Select

Select

Expl

orat

ory

phas

eCo

nfirm

ator

yph

ase

Assay / stat analysis / select BMs

Background variability

Thresholds

Assay availability / development

DILI BM step 4 list

Qualification

Assay / stat analysis / select BMs

SAFE-T Biomarker qualification processElements and process flow

Q2 2009

Q1 2010

Q2 2011

Q2 2014

10

Biomarker identification

Assay identification

Required sample sizes

Target populations

Biomarker qualification

Pathologies of interest

Assay development

Database costs

Investigative sites

Number and design of clinical studies

Capabilities

EFPIA studies with add-on sampling

Prospective studies Retrospective samples

Incidence of target disease

Sampling and shipment processes

Sample matrices and volumes

Drug selection

Data capture, management, analysis

Number of study sites needed

Number of patients needed

Biomarker submission

Recruitment ratesData management resources

Extent of in-kind support required

Type of in-kind support required

Biomarker qualification processInitial challenges

Technology platform

Data entry resources

11

Key achievements at project half-time

• Biomarker candidates prioritised, assay development well advanced

• Central biobank for sample storage up and running

• Database and data capture system up and running

• Academic sites: 17 prospective clinical studies initiated

• EFPIA partners:

– Completed SAFE-T studies: 2

– Retrospective samples: >6500 patients from 4 studies

– Ongoing add-on sampling: 3 studies

– Submitted or under preparation: 5 studies

• Initiated regulatory interactions via briefing meetings with EMA/FDA

• Established collaboration with Predictive Safety Testing Consortium (PSTC)

12

Clinical partners • Patient / sample codes

• Guidelines for sample collection and processing, shipment instructions

Samples

Biomarker assay group

Sample request

Sample approval team

Data analysis group

Patient – sample information

Regulatory requirements group

29 studies planned

1538 aliquots

SOPs, informed consent

10 studies, >60,000 aliquots

Samples

SAFE-T biobank: up and running

Approval of sample release

13

SAFE-T database: up and running

14

DILI biomarkers – status of assay developmentCandidate biomarker

miRNA 122albumin mRNAMicroglobulin precursor (Ambp) mRNA

High mobility group box 1 (acetylated vs. non-acetylated)Conjugated/unconjugated bile acids

High mobility group box 1 (acetylated vs. non-acetylated)ALT 1 & 2, isoform specificF-protein (HPPD)Arginase 1Keratin 18 (caspase cleaved & intact)Alpha fetoprotein (AFP)Regucalcin (RGN)Glutathione S-Transferase (GST-alpha)ST6gal IOsteopontinColony stimulating factor receptor (CSF1R)Paraoxonase 1 (PON1)ProthrombinLECT2

Glutamate dehydrogenase (GLUD, GLDH)Purine nucleoside phosphorylase (PNP)Malate dehydrogenase (MDH)Sorbitol dehydrogenase (SDH)ALT1/2, isoform specificAc

tivity

ass

ayRN

ALC

MS

Imm

unoa

ssay

Status

Ready for sample screeningReady for small sample sizesOptimization phaseIn developmentDevelopment necessary

15

HMGB1 and Cytokeratin 18Mechanism based biomarkers

Apoptosis:• Keratin‐18 – intermediate 

filament protein / structural integrity

• Is cleared by caspases• Fragment released into blood• Full length K18 passively released 

during necrosis

Necrosis and Inflammation:• HMGB1 – chromatin binding protein• Passive released by necrotic cells• Active released by activated immune cells 

(hyper‐acetylated (Lys NLS))• Cytokine activity (TLR/RAGE)

Antoine DJ et al., 2010 Mol MedAntoine DJ et al., 2009 Toxicol Sci

Slide courtesy Neil French, MRC CDSS

16

Patients post acetaminophen overdoseMarkers for inflammation, necrosis, and apoptosis

Association with King‘s College Criteria Based on Antoine DJ et al., 2012 J Hepat

• Acetylated HMGB1 may be a prognostic DILI marker, indicating extent of inflammation• Caspase cleaved cytokeratin 18 may have value as a prognostic DILI marker, indicating

involvement of apoptosis as protective mechanism

17

Parallel to qualification: DILI biomarker discovery

Why?• Biomarker candidates do not cover all objectives of SAFE-T DILI WP

– Lack of susceptibility markers

– Lack of sensitive functional markers, some pathologies poorly represented

– Most markers identified in pre-clinical models

How?• Based on human DILI cases from SAFE-T clinical studies

• Characteristic changes in serum proteome and metabolome expected– Mass spec and protein antibody array analyses of plasma samples planned

• Genetic analysis not planned, but possible collaboration with iDILIC

IMI SAFE-

T

CollaborationKey to success

19

PSTC

SAFE-T

Biomarker qualification

fNIH

SAEC

DILIN

? ? ? ?

• SAFE-T is collaborating closely with C-Path‘s Predictive Safety TestingConsortium (PSTC), utilizing synergies and preventing overlaps

• There may be more opportunities to expand collaboration, helping toincrease efficiency and maximize output

Conclusions

• Qualification of new safety biomarkers can best be done in a setting of large scale pre-competitive collaborations such as the IMI-SAFE-T consortium, PSTC, and others alike

• The IMI SAFE-T consortium has made significant progress during thepast 2.5 years

• Consortium systems and processes for sample collection, processing, storage, shipment, and analysis have been set up and are running well

• Data capture, storage, management, and analysis tools are in place

• Seventeen prospective clinical studies have been initiated, but need toincrease recruitment

• SAFE-T may serve as an encouraging example to establish furtherprecompetitive collaborations focusing on drug safety

20

Kevin ParkNeil FrenchDaniel Antoine

Thomas Joos

Hannes PlanatscherJens GoepfertNicole Schneiderhan-Marra

Teresa PadroLina Badimon

AcknowledgementsEric Massana Kaïdre Bendjama Nicolas von Behr David Reynolds Vicky McGrath Hugo Hämmerle Rolf Seeland Ronni Gamzu

Pere Berga Marti Peter Thomann Heidrun Ellinger-Ziegelbauer Robin Privman Barry Hayes Günter Beck Theresa Klein Bernd Stowasser

Mariona Auli Attila Garami Matthias Gottwald Dominique Brees Rory Connolly Thomas Schweikert Thomas Berg Isabelle Clavier

Silvia Lopez Béatrice Molac Susanne Schwenke Ceri Collins Mark Pinches Jens Göpfert Florian van Bömmel Frederic Galli

Jose-Luis Diaz Fuat Firat Katja Matheis Erik Kuja David Brott Stefanie Rimmele Stefanie Frank Joachim Tillner

James Matcham Stella Suzanne Christine Rentzsch Andrew Zehner Ina Schuppe Koistinen Hannes Planatscher Sabine Boas-Knoop Xavier Benain

Alexandre Mencik Brice Suire Uwe Bamberger Shelli Schomaker Håkan Andersson Frank Dieterle Lina Badimon Joost Dwerhagen

Simon Gibbs Barbara Fischer Birgit Stierstorfer Cornelia Ciorciaro Anders Samuelsson Peter Hoffmann Teresa Padro Isabelle Pirani

Patrice Cacoub Dorina Bratfalean Arno Kalkuhl Laura Suter Dick Sally Price Michael Merz Xavier Sánchez-Vallve Catherine Lunven

David Saadoun John Haselden Joachim Stangier Piotr Szczesny Jesper Hedberg Robert Schmouder Nuria Nadal Olivier Pasquier

Damien Sene Andrew Nicholls Kristiane Wetzel Barbara Lenz Björn Glinghammar Ursula Knauf Gemma Vilahur Jean-Charles Gautier

Michel Buchert Elaine A. Irving Dr. Volker Mahlbacher Jacky Vonderscher Pierre Dönnes John Prince Silvia Gómez Nathalie Piton

Anne Skrobot Vincent Mooser Dorothee Schwall-Rudoph Lucette Doessegger Terry Reed Jeffrey Donohue Sandrine Schwartz Marie-Hélène Pascual

Florence Ghrenassia Fiona J McClure Marcus Kostka Manfred Argast Ann-Cathrine Jonsson- Francois Legay Daniel Sanchez Muriel Breitbach

Thierry Poynard Aubrey Swain Nadine Erne Christoph Wandel Li-Ming Gan David Laurie Thomas Joos Sylvie Brohier

Mona Munteanu Theo Dare Andreas Port Rodolfo Gasser Jenny McKay Alexandre Avrameas Martina Kahnert Jean-François Gallas

Hugo Perazzo Ian Roman Ulf Neumann Bernhard Risse Jonas Bergström Marie Anne Valentin Jessica Mittelstädt Reinhold Stockbrugger

Servane de Penquer Federica Crivellente Maximilian Schmeding Rosemarie Kientsch-Engel Marcus Gry Philip Bentley Manfred Schmolz Bernard Levin

Laurent Becquemont David Howe Eveline Fräßdorf Thomas Schindler Harry Southworth Gerd Kullak-Ublick Thomas Knorpp Ali Yağız ÜRESİN

Benoit Labarthe Landry Cochard Janine Heyder Isabelle Gilet Scott Adler Franck Meyer Derek Leishman Hugh Laverty

Anne Gysembergh-Houal Marc Loher Volker Schmitz Nadir Arber Sif Ormarsdottir Steve Hall Sarah Chatham Kevin Park

Joe Keenan Christiane Andriamandroso Eckart Schott Sara Kraus Stephen Furlong Stefan Sultana Simon Harper Munir Pirmohamed

Camilla Stephens Maribel Lucena Patrick Murray Colin Green Magnus Nord Michael Lawton Paul Commander Neil French

Brooke Bergeron Jean-Marc Vidal Marisa Papaluca Raúl Andrade

21