Pierre Fabre Immunology Centre Oncologique Pharmacologie …€¦ · Atelier GPCO Nice 22 Novembre 2013 Centre d’Immunologie Pierre Fabre 1 Pierre Fabre Immunology Centre Centre

Centre d’Immunologie Pierre Fabre 1 Atelier GPCO Nice 22 Novembre 2013

Pierre Fabre Immunology Centre

Centre d’Immunologie Pierre Fabre 1

Dr. CORVAIA Nathalie

Managing Director

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CIPF

Innovation : discovery, development and production of Mabs and ADC for

Oncology

Production unit in an R&D integrated site (2011)

IGF-1R/

Dalotuzumab

C-Met/

CXCR4/

hz515H7

1 Mab - CIPF (MERCK -2004)

3 Mab - CIPF (Pierre Fabre – 2013)

2 Mab - CIPF (ABBOTT - 2010)

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2013 itolizumab(CD6)[Ind] ado trastuzumab emtansine(HER2) obinutuzumab[low Fuc] (CD20) [Remsima /”infliximab” EMA]

2012 mogamulizumab[low Fuc](CCR4)[Jpn] pertuzumab(HER2) ziv-aflibercept[PrFc]

(VEGF) raxibacumab(antrax)

2011 brentuximab vedotin[IgG1-vcMMAE](CD30) belatacept[PrFc]

(CD80/86) aflibercept[PrFc](VEGF)

belimumab(BLysS) ipilimumab(CTLA-4)

2010 denosumab[IgG2](RANK-L)

2009 golimumab(TNFa) catumaxomab(EpCAM/CD3) ustekinumab(IL12/23) canakinumab(IL1) ofatumumab(CD20)

2008 rinolacept[PrFc](IL1) certolizumab[Fab-PEG]

(TNFa) romiplostim[FcPe](TPO) [Clotinab/”abciximab” So-Ko]

2007 eculizumab[IgG2/4](C5) [Reditux/”rituximab” Ind]

2006 ranibizumab[Fab](VEGFA) panitumumab[IgG2]

(EGFR)

2005 nimotuzumab[Chi](EGFR) abatacept[PrFc](CD80/86) tocilizumab(IL6R)[Jpn]

2004 cetuximab(EGFR) bevacizumab(VEGFA) natalizumab[IgG4](a4 integr)

2003 131I-tositumomab[mIgG2a](CD20) omalizumab(IgE-Fc) efalizumab(CD11a)[withdrawn, 2009] alefacept[PrFc]

(CD2)

2002 111In/90Y-ibritumomab tiuxetan[mIgG1](CD20) adalimumab(TNFa)

2001 alemtuzumab(CD52)

2000 gemtuzumab ozogamicin[IgG4-calicheamycin](CD33) )[withdrawn, 2010]

1999 1998 basiliximab(CD25) palivizumab(RSV-F) infliximab(TNFa) trastuzumab(HER2) etanercept[PrFc]

(TNFa)

Technologies 1997 rituximab(CD20) daclizumab(CD25)

Transgenic mice (10Y) 1996 1995 edrecolomab[mIgG2a]

(EpCAM) [Ger, withdrawn]

1994 abciximab[Fab](GPIIb)

Phage display (9Y) 1993 Humanization (11Y) 1986 muromomab[mIgG2a]

(CD3)

Chimerization (10 Y) 1984 Nobel Prize for mAbs

Approved therapeutic Antibodies & related-products (INN= International Non-proprietary Names, WHO)

* FDA, EMA, SFDA and /or DCGI (SFDA = China FDA;

DCGI = Drugs Controller General of India)

40 mAbs, Fabs, Fc-fusions, ADCs, RadioIC, Bispec (27 y)

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Infliximab (7.5 B.) Rituximab (7.1 B.) Cetuximab (1.9 B.)

2012 sales, La Merie, Business Intelligence, May 2013

Trastuzumab (6.3 B.)

Bevacizumab (6.1 B.)

Ranibizumab (4.0 B.) (Fab)

Natalizumab (1.6 B.)

Omalizumab (1.3 B.)

Eculizumab (1.3 B.)

Palivizumab (1.0 B.)

Adalimumab (9.5 B.)

Ustekinumab (9.0 B.)

Fc-fusion proteins [2]

Etanercept (8.4 B.)

Abatacept (1.2 B.)

Best selling (> 1 billion USD)

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Strategies to develop news Mabs ?

• Choice of targets

• Validated or innovative targets

• Immuno-oncology

• Mab format

• ADCs, New scafolds, Bispecific antibodies

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• Targeted approaches: membrane targets (overexpressed) or circulated ligands

(1) Clinically validated: 2nd and 3rd generation mAbs

• CD20, Her2, EGFR, VEGFA, EpCAM

(2) Pre-clinically validated and ongoing clinical trials

• IGF1-R, IGF1/2, HGF, c-Met, Her3, VEGF ou VEGF-R,

Trail-R, IL6 ou IL6R, IL4/IL13, CD19, CD22, CD30,

CD44, CD80, CD151, CTLA-4, ICAM-1

• Fonctional approaches: potential new target

• Fonctional screening

• Mab Selection and Ag identification • RAAG12, CD9, JAM-A, TSN-1

Pre-validated targets

Innovative targets

• Competition

• Safety

• POC

• IP

Validated targets

Target selection: strategy and challenges

Beck A, Wurch T, Bailly C & Corvaïa N. Nature Rev Immunol (Mai 2010)

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Challenges in developing new Mabs against innovative targets

• Target expression

• Tumor versus normal

• Target validation

• Major role in cancer progression

• Addiction

• Mechanism of action and biomarkers for patient selection ?

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Mab target identification platform

WB

IP/WB Proteomics

siRNA

Protein array

Identification

Validation

6F4 Mab : JAM-A

Tumor

cell lines

312F12 Mab : undisclosed

target

Tumor

tissues

Immunizations

Mab Generation

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Functional

selection

In vitro cell

proliferation

inhibition

MCF-7 model

In vivo

anti-tumor activity Mice

Immunization

Target identification

Proteomic

approach Tumor cells

as immunogens

Target of the mAb 6F4: JAM-A JAM-A = Junctional adhesion molecule A

characterized by two Ig-like domains: a membrane distal variable immunoglobulin-like loop (D1) and a membrane proximal constant immunoglobulin-like loop (D2)

Constituent of TJ: cell polarity involved in cell adhesion, migration

Functional approaches (1)

JAM-A identified as potential anti-tumor

target

(patent filled November 2006, Goetsch et al Int J

Cancer. 2013 Mar 15;132(6):1463-74 )

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• Human JAM-A is a potential target in oncology

• JAM-A is over-expressed on tumor cells vs. normal cells

• Anti-JAM-A Mab inhibits tumor growth in vivo through

• Inhibition of cell proliferation

• Down regulation or shedding of the molecule

• Down regulation of genes involved in translation machinery

• First humanized antibody targeting JAM-A available

• Mechanism of action ? : biomarkers for patient selection ?


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Generation of resistant A549

•Tri-therapy treatment

Antibody screening by IHC

• On frozen sections

• Antibodies were selected when:

Non treated A549 were negative

Resistant A549 were positive

Immunisation

• Mice Tolerization with non treated A549

• Immunisation of toleri-zed mice with resistant A549

• Fusion and antibody production

2E11 selection

Non treated A549 Resistant A549


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2E11 staining

Resistant tumors

* *

*

*

No staining with 2E11

Untreated tumors Proteomic approach

Maldi-MS analysis

TSN-1 is the target

Confirmation by western blot

and SiRNA approaches

Target identification: IHC and in vivo activity

25

150

100 75

50

37

20

25

150

100 75

50

37

20

WB

2E1

1

WB

MO

1(C

om

me

rcia

l Ab

)

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Tetraspanin-1: TSN-1

• 2E11 recognizes tetraspanin-1 (EC2) • Protein structure (241 AA)

• Location 1p34.1

• 241 AA

• Structure

» 2 ECDs

» 4 TMDs

» 2 ICDs

• Tetraspanin Functions • Cell-cell interactions

• Adhesion

• Migration

MDAH-2774 OVCAR-3

WT-Tspan-1 EGFP WT-Tspan-1 EGFP

4xNA-TSPAN-1 EGFP 4xNA-TSPAN-1 EGFP

4 N-glycosylation sites

Tumor development CD9 (ARIUS)

CD63 (ARIUS; licensed to Genentech)

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Role in tumorigenesis ? TSN-1 expression is correlated with Tumor grade

Need to address the role of TSN-1 in cancer development

Need to know more about TSN-1 distribution

Tumors TSN-1 expression Method

Cervical Carcinomas

Hepatocellular carcinoma

Ovarian Cancers

Breast Tumors

- Up-regulation of TSN-1 is associated with carcinogenesis

- Association of NET-1 gene expression with human hepatocellular carcinoma

- Labeling intensity is correlated with carcinoma type and could be used as diagnostic marker (intracellular)

- No expression in normal Breast. Up regulation in breast tumors

Int. J. Cancer 2002

Int J Surg Pathol. 2007

Cancer Letters in press

Int. J. Cancer 2007

Xenopus - Tetraspanin-1 regulates gastrulation movements and neural differentiation in the early Xenopus embryo

Differentiation 2006

Midkine - Midkine interacted with TSN-1 and and facilitated TSN-1 association with a6b4 BBRC 2008

Target Identification: TSN-1 and cancer

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• Human TSN-1 is a potential target in oncology

• TSN-1 is over-expressed on tumor cells vs. normal cells

• Anti-TSN-1 Mab inhibits tumor growth in vivo

• Tetraspanins are an emerging class of targets in Oncology

• Mechanism of action ? : biomarkers for patient selection ?

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G protein-coupled receptors (GPCR)

• GPCR family

• About 800 known members

• 370 GPCR as potential targets for drug development

• 60 are targeted by marketed small molecules

• Less than 25 are targeted by bio-therapeutics

22% of them could be good targets in Oncology

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GPRC families and structure

• 3 families

• Family A : ex rhodopsin. Ligands are peptides, neuripeptides, …

• Family B : secretin and adhesion families. Bind large peptides

• Family C: glutamate family

• 7TMD

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Challenges for the generation of anti-GPCR Mabs

• Low cell surface expression

• Mab screening

• Mab access

• No purified protein available for immunisation

• Peptides

• Cell extracts

• Transfected cell lines

• Difficulties to obtain functionnal Mabs : need several strategies

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(A) Direct effect by antigen binding (Fab/CDRs): - inhibition of

- CXCR4/SDF-1 signaling - Cell migration - Cell invasion - Cell survival

- Inhibition of angiogenesis - Induction of

- Cell mobilization in Cynomolgus monkey

Anti-CXCR4 hz515H7 mAb : dual effect

(B) Effector functions (Fc binding) - Antibody-Dependent Cell mediated Cytotoxicity - Complement-Dependent Cytotoxicity cell lysis

IgG1 hz515H7

Potential best in class molecule

CXCR4

Target Cell

SDF-1

C1q

Effector Cells (Natural Killer, macrophage)

FcRIIIa

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Functional Sustainability Innovation

Antibody Biotech Unit

Plebiscited

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• Immuno-oncology

• Mab format


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Ipilimumab (Yervoy) 2011 in metastatic melanoma

From Ott et al, clinical cancer res 2013, 19, 5300

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Promising field

« Immunotherapy will likely form the

backbone of 60% of all cancer

treatment in 10 years »

A.S. Baum, Citi, May 2013

« It is certainly the biggest change in

oncology that we’ve seen » about BMS

and Merck drugs

T. Butler, Barclays, June 2013

« Immunotherapy has become the dominant story in

pharma, with impressive responses and survival data in

some of the most intractable cancers»

Goldman Sachs August 2013

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Immune checkpoints : multiple potentiel targets

From Ott et al, clinical cancer res 2013, 19, 5300

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Emerging class of molecules

From Pardoll Nature rev. Cancer 2012, 12, 252

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• Immuno-oncology

• Mab format


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Specfic targeting du to Mab

Cytotoxic molecule

Linkers for drug conjugation

Antibody drug conjugates

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Cytotoxic warheads for ADCs

• Calicheamicin (Wyeth/Pfizer): • Mylotarg® approved 2000, withdrawn 2010

• Auristatin E/F (Seattle Genetics): • Adcetris™ FDA-approved 2011

• Maytansinoids (ImmunoGen): • Kadcyla™ FDA-approved Aug 2013

• Duocarmycins (Medarex/BMS and Synthon): • No clinical data yet

• Anthracyclines (Immunomedics): Phase I • Camptothecins (Immunomedics): Phase I • Indolino-benzodiazepines (ImmunoGen) • Pyrrolobenzodiazepines

• Spirogen/ADC Therapeutics, Sanofi, Seagen, Genentech

• Auristatin E/F analogs (Ambrx, Pfizer, Bayer, Mersana) • Amanitin (Heidelberg Pharma)

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MylotargTM (2000-2010)

O

O

O

IS

O

OHO

HO

O

HO

ONH

O

HO

O

HO O

HN

O

OS

SNH

O

N

OO

NH

O

ON

O

O

O

Calicheamycin =

(2 to 3/ IgG)

Gemtuzumab

(HzIgG4SerPro)

-Lys-NH2 (random)

Linker

-hydrazone-

Hughes B, Nature Drug Discovery 2010

FDA approval: 2000

Market withdraw: 2010

Gemtuzumab ozogamicin

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2nd ADCs generation reaching the market

(A) Kadcyla (ado trastruzumab emtansine): 2013

=> Lys, maytansinoids (SMCC/SPP-DM1, SPDB/sulfo-DM4), ImmunoGen

=> 12 in clinical trials (31%)

(B) Adcetris (brentuximab vedotin): 2011

=> Cys, auristatins (mcMMAE, vcMMAF), Seattle Genetics

=> 21 in clinical trials (54%)

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ImmunoGen tech: 12/39 in clinical trials (31%)

Maytansinoids (12) Target Drug+ linker Developer Stage Indication (1) Ado trastuzumab emtansine(Kadcyla) HER2 SMCC-DM1 Roche/ Genentech 2013 HER2-positive mbreast cancer (2) Lorvotuzumab mertansine, IMGN901 CD56 SPP-DM1* ImmunoGen Inc. Ph II MM, Merkel Cell Carc, Ovarian (3) SAR3419 CD19 SPDB-DM4* Sanofi-Aventis Ph II NHL, B-ALL (4) SAR566658 CA6 SPDB-DM4* Sanofi-Aventis Ph I Breast, ov., cerv., lung, pancreatic (5) Indatuximab ravtansine, BT-062 (chIgG4) CD138 SPDB-DM4* Biotest/ ImmunoG Ph I Multiple myeloma (6) IMGN-289 EGFR SMCC-DM1 ImmunoGen Ph I NSCLC, SCCHN (7) IMGN-388 Integrin av SPDB-DM4* ImmunoGen Ph I Solid tumors (8) IMGN-529 CD37 SMCC-DM1 ImmunoGen Ph I NHL (9) IMGN-853 FolR1 sSPDB-DM4* ImmunoGen Ph I Ovarian, NSCLC (10) BAY 94-9343 Mesothelin SPDB-DM4* Bayer HealthCare Ph I Mesothelin-expressing tumors (11) AMG-595 EGFRvIII SMCC-DM1 Amgen Ph I Recurrent Gliomas (12) AMG-172 CD70 SMCC-DM1 Amgen Ph I Renal cancer Cleavable/ Non-cleavable ratio: 7/5 ; – Hematologic/ Solid tumors ratio: 3/9 DM1 = ; DM4 = ; SMCC=, SPP = SPDB = Sulfo-SPDB Lambert J, BJCP 2013 Beck A, Carter P et al, mAbs 2013

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Seagen tech: 21/39 in clinical trials (54%)

MMAE/F Target Drug+ linker Developer Stage Indication (1) Brentuximab vedotin (AdcetrisTM) CD30 vcMMAE SeaGen/ Takeda 2011 ALCC, HL (2) Glembatumumab vedotin (huIgG2) GPNMB vcMMAE Celldex Ph II Breast cancer, melanoma (3) PSMA ADC PSMA vcMMAE Progenics/ SeaGen Ph II Prostate cancer (4) Pinatuzumab vedotin, RG7593 CD22 vcMMAE Genentech/ Roche Ph II NHL (5) Polatuzumab vedotin, RG7596 CD79b vcMMAE Genentech/ Roche Ph II NHL (6) Vorsetuzumab mafodotin, SGN-75 CD70 mcMMAF Seattle Genetics Ph Ib NHL, RCC (7) SGN-CD19A CD19 mcMMAF Seattle Genetics Ph I B-cell non-Hodgkin lymphoma (8) SGN-LIV1A LIV-1A vcMMAE Seattle Genetics Ph I Breast (9) AGS-5ME (huIgG2) SLC44A4 vcMMAE Agensys/Astellas Ph I Prostate, Pancreatic, Gastric (10) AGS-22ME Nectin-4 vcMMAE Agensys/Astellas Ph I Solid tumors (11) AGS-16M8F (huIgG2) AGS-16/ENPP3 mcMMAF Agensys/Astellas Ph I Renal cell carcinoma (12) MLN0264 GCC vcMMAE Millenium (Takeda) Ph I Gastrointestinal malignancies (13) RG7450 (DSTP3086S) STEAP1 vcMMAE Genentech/ Roche Ph I Prostate (14) RG7458 (DMUC5754A) MUC16 vcMMAE Genentech/ Roche Ph I Ovarian (15) RG7599 (DNIB0600A) NaPi2b vcMMAE Genentech/ Roche Ph I NSCLC, ovarian (16) RG7598 ? vcMMAE Genentech/ Roche Ph I Multiple myeloma (17) RG7600 ? vcMMAE Genentech/ Roche Ph I Pancreatic, Ovarian (18) RG7636 ETBR vcMMAE Genentech/ Roche Ph I Melanoma (19) ABT-414 EGFRvIII Auristatin Abbvie Ph I/II Squamous Cell Tumors (20) Anti-5T4 5T4 mcMMAF Pfiser Ph I Solid tumors (21) HuMax-TF-ADC TF vcMMAE Genmab Ph I Multiple solid tumors Cleavable/ Non-cleavable ratio: 14/3 (3 non-disclosed) – Hematologic/ Solid tumors ratio: 5/15 - IgG1/ IgG2 ratio: 17/3 GCC = Guanylyl Cyclase C

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Misceleanous: 6/39 in clinical trials (15%)

Calicheamycin (2) Target Drug+ linker Developer Stage Indication (1) Gemtuzumag ozog, Mylotarg (IgG4) CD33 hydrazone-CM1 Pfizer 2000-2010 (US) AML (2) Intotuzumab ozogamicin (IgG4) CD22 hydrazone-CM1 Pfizer PhIII NHL Doxorubicin (hLL1) (1) Target Drug+ linker Developper Stage Indication (1) Milatuzumab doxorubicin CD74 hydrazone Immunomedics Ph I/II Mulitple myeloma SN38 (irinotecan prodrug) (2) Target Drug+ linker Developer Stage Indication (1) Labetuzumab-SN38 CEACAM irinotecan Immunomedics Ph I Colorectal Cancer (2) IMMU-132 TROP2 irinotecan Immunomedics Ph I Solid tumor PBDs (1) Target Drug+ linker Developer Stage Indication (1) SGN-CD33A CD33 PBD (EC-mAb) Seagen (Spirogen) Ph I AML

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• High specific for a target

Natural products Monoclonal Antibody

IGF1-R, cancers du sein, colon (phase 2)

• Small molecule inhibitors from plant extracts • Potent anti-tumor activity • Lack of selectivity

JAVLOR (vinflunine) in bladder cancer

NH

N

F

N

CH3

H

HO

CH3

N

CH3 O

O

O

CH3

O

CH3

O

O

CH3

OH

CH3

FH

cMET, tumeurs solides (phase 1)

Antibody drug conjugates

Cellule tumorale

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Conclusions

Innovative targets, immunoOncology field and ADC are new challenges for treating patients

Biomarker identification and development will guide to select the best patient population to treat

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Thanks Experimental Oncology

L. Goetsch,

Biochimistry

JF Haeuw

Molecular and Cellular Biology

M. Tesar

Physico-chemistry

A. Beck

Industrial Pilot

O. Cochet

CQ

S. Lauthier

120 people dedicated to R,D and production

Of Mabs

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Documents

Pierre Fabre Immunology Centre Oncologique Pharmacologie …€¦ · Atelier GPCO Nice 22 Novembre 2013 Centre d’Immunologie Pierre Fabre 1 Pierre Fabre Immunology Centre Centre