Sym001 Rozrolimupabfor treatment of Immune Thrombocytopenic Purpura (ITP) and Anti-D Prophylaxis (ADP)

The Sixth Plasma Product Biotechnology Meeting

Torben P. Frandsen, Director of Antibody Chemistry

Symphogen A/S

3rd generation of therapeutic antibodies

Diverse: + Specific: ÷

Diverse: ÷Specific: +

Diverse: +Specific: +

Immunoglobulins

Monoclonal antibodies

Recombinantpolyclonal antibodyproducts

Symplex™ Discovery engine

• Human antibody drug lead candidates

– Natural VH-VL pairing preserved

– Natural high affinities and reactivities

Sympress™ Expression engine

• Recombinant polyclonal Ab products

• Single batch manufacturing

• Batch-to-batch consistency

• Industrial scale

18 M

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s to Precln

ical Develo

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Proprietary antibody technologies

http://www.symphogen.com/web/guest/symplex

http://www.symphogen.com/web/guest/sympress

Symphogen product pipeline

Sym001 rozrolimupab

Indications ITP: Treatment of Immune Thrombocytopenic Purpura

ADP: Anti-D prophylaxis to prevent Hemolytic Disease in Newborns (HDN)

Target Rhesus D on Red Blood Cells

Opportunity Replacement of blood-derived immune globulin products(IVIG and Anti-D)

Class Recombinant polyclonal antibody product (pAb)

Composition 25 human full-length IgG1 Abs with documented RhDreactivity

Stage ITP: Phase 2ADP: Phase 1/2, RBC challenge in healthy volunteers

Sym001 product characteristics

• Sym001 antibody leads identified from Danish anti-D donors by phage display technology

• Sym001 comprises 25 unique human IgG1 antibodies

• Sym001 comprises antibodies with verified reactivity against RhD III, RhD IV, RhD VI, RhD VII

• Sym001 clones do not react with: ABO, E, f, P1, Lea, Leb, Lub, N, S, s, JKa, JKb, Fya, Fyb, K, k, Kpb, Rd, Vel, Ge, and H(0).

• Sym001 mimicks the IgG repertiore of anti-D donors

Sym001 mimics the IgG repertoire of anti-D donors

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JH gene usage

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(A) frequencies of heavy chain V gene family (B), VH3 gene allelic usage (C), J gene segment usage and (D) CDR3 region length in amino acids.Sym001 (white bars) is compared to a compilation of the 165 known VH sequences of human anti-RhD antibodies (grey bars).

Sym001 mediates erythrocytedestruction in vitro• Several MoA have been described for anti-D in RhD prophylaxis

• Destruction of RhD+ erythrocytes is the most likely MoA;

– Phagocytosis

– Antibody-dependent cellular cytotoxicity (ADCC)

• In vitro efficacy of Sym001 is similar to anti-D products for binding potency assays

Sym001 blocks platelet uptake in macrophages

ITP model

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Concentration of anti-D (ng/ml)

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Sym001-rWS1WinRho

• Anti-D MoA for inhibition of platelet destruction in ITP:

– Anti-D opsonized RhD+ erythrocytes competes for FcγR at RES macrophages– Anti-D opsonized RhD+ erythrocytes induce refractory period in RES macrophages

• Sym001 blocks platelet uptake in macrophages

Manufacturing, Release and Characterization of RecombinantPolyclonal Antibody Products

SympressTM Technology

Polyclonal antibody expression

• Mammalian expression (CHO)

• Site-specific integration (Flp-In system)

• Single integration event per cell

• Homogenous expression & growth

• Identical constant regions

Generation of the PALS cell lines

• Identical sub-cloned parental cells • Identical plasmids, apart from the antibody genes• Transfection, adaptation and banking same for all PALS• Each cell produces only one antibody• PALS generation a one time event

GMP cell banks and manufacturingprocess

• The pMCB and pWCB are the GMP cell banks

• Both are polyclonal cell banks

• One vial of pWCB is thawed for each Sym001 DS batch

Structural integrity

• CIEX profiling• Identification of marker peptides

by LC-MS

Sym001

Assessment of polyclonality

Structural integrity

Analysis of constant region

• Western blotting• Capillary electrophoresis• Size exclusion chromatography

• Peptide mapping• Disulphide bridges• Carbohydrate analysis

Antibody characterization strategy

Analysis of variable & constant region

Typical characterization of mAb

• Western blotting• Capillary electrophoresis• Size exclusion chromatography

• Peptide mapping• Disulphide bridges• Carbohydrate analysis

Release and characterization assays

Release assays• Identity

– Constant region

– Sym001 specific

• Purity

– Product related

– Process related

• Quantity

• Potency

• General

Characterization assays• N-linked carbohydrate

• Primary structure

• Secondary structure

• Thermal stability

• Product related contaminants

• Biological characterization

• In vitro safety profiling

Manufacturing of Sym001

Sym001 Batch Cell bank Scale (L) Use

MYS05-01 pMCB 400 Experimental

MYS05-02 pMCB 400 Toxicology studies

MYS05-03 pMCB 400

MYS05-04 pMCB 400

MYS08-01 pWCB (pWCB-02) 400

MYS08-02 pWCB (pWCB-02) 400

MYS08-03 pWCB (pWCB-02) 400

GMP campaign intended for future clinical studies

GMP campaign for clinical phase 1 and 2

Working standardTox batchGMP 1 batchGMP 2 batch

Similar compositional diversity of Sym001 demonstrated in 4 manufacturing runs

CIEX profiling. Spiking with RhD mAb

m i n1 0 20 3 0 40 5 0 60 7 0 80 9 0 1 00

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Working standardRhD240Working standard spiked with RhD240

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m A U

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CIEX profiling. Spiking with RhD mAb

RhD157 Working standardRhD157Working standard spiked with RhD157

CIEX profiling

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Marker Peptide Method

SaM16959 11 05061502:11_UV SaM16959 11 05061502:11_Fractions SaM16959 11 05061502:11_Inject

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Drug Substance HC/LC Separation Protease Digestion

HPLC SeparationMS AnalysisAb Identification

Development of a marker peptide method

The presence of all 25 antibodies have been confirmed in 4 scale-up batches

Sym001 CMC development and characterization• Regulatory path for future approval of Sym001 established• Two-tier polyclonal cell bank concept established

(pMCB/pWCB) to support manufacturing during productlifespan

• Recombinant polyclonal antibodies can be reproducibly manufactured at industrial scale– 400 L scale– 12,500 L simulation

• An extensive package of methods for release and characterization of recombinant polyclonal antibodies have been developed

• Methods to control consistency have been developed

Indications and clinical development

Hemolytic Disease of the Newborn

• 16% of caucasians lack RhD on their red blood cells (RhD-)

• Annually there are 1.3 million live births to RhD-women in the US and Europe

• HDN risk if a RhD- woman is pregnant with RhD+ fetus

– RhD+ fetal blood may leak to mother’s circulation causing a maternal immune response to RhD antigen (isoimmunization)

• Future pregnancies

– Mother’s immune response causes destruction of red blood cells in a RhD+ fetus and development of HDN

• HDN

– Fetal hemolysis and multi organ failure – Severe handicaps and death

RhD-

RhD+

Immune Thrombocytopenic Purpura (ITP)Rare autoimmune disease resulting in platelet destruction

• Children: Acute disease 2-3 weeks after viral infection. Spontaneous remission in 80% within 6 months

• Adults: Chronic disease withinsidious onset. Secondary to viral infections, drugs and malignancies

• New ITP cases annually: 19,000 adults and 8,000 children (US and Europe)

Bruises and bleedingsPlatelets/mm3:< 30,000 Treatment< 20,000 Spontaneous

bleeding<10,000 Epistaxi,

hematuriableedings frommucosa and skin

< 5,000 Spontaneousinternalbleedings

• 7 dose cohorts

Sym001 – phase 1

Hjelmstrøm et al. ASH 2008

Cohort 17 RhD+ subjects

Cohort 27 RhD+ subjects

Cohort 3A9 RhD+ subjects

Cohort 4A9 RhD+ subjects

Cohort 59 RhD+ subjects

Cohort 69 RhD+ subjects

Cohort 3B9 RhD negative subjects

Cohort 4B9 RhD negative subjects

Cohort 79 RhD+ subjects

Confidential /28

Hemoglobin (mean change from baseline per cohort in RhD+ subjects)

Hjelmstrøm et al. ASH 2008

ADP: Proof of mechanism study

• Sym001 given to 24 RhD- negative healthy males challenged with RhD+ RBCs i.v.

• Positive control: Rhophylac® given to 12 RhD- negative healthy males challenged with RhD+ RBCs i.v.

• Study objectives: – To assess the ability of Sym001 to eliminate

RhD+ red blood cells in RhD- subjects– To assess suppression of RhD immunization

• Preliminary results: Dose-dependent clearance of RhD+ RBCs established with Sym001at five days(Press release 21 Nov 2008)

ITP: Proof of concept Study

• Initiated Jun ’08 (Press release 16 Jul 2008)

• Phase 2 dose-escalation study

• Objective:

– To evaluate safety

– To determine pre-liminary efficacy on platelet count

• 23 sites in Europe

Conclusions

• Sym001 is expected to have same or better benefit/risk and coverage as blood-derived anti-D while offering recombinant benefits over IVIG and anti-D: – Safety, convenience, supply, drug consistency

• Recombinant polyclonal antibodies can be reproducibly manufactured at industrial scale

• Recombinant polyclonal antibodies are well characterized through an extensive package of methods for release and characterization

• Clinical data shows good safety profile. POC data from phase 1 and phase 2 in RhD+ and RhD- subjects

• Regulatory expectations mapped through close dialogue withauthorities in US and Europe

Acknowledgements

• Sym001 is co-developed with Biovitrum

• Numerous employees at Biovitrum & Symphogen

• Contact: tpf@symphogen.com

• Thank you