Embracing Complexity: Understanding IVIg to Rationally Design Novel Therapeutics

Anthony M. Manning, PhD

VP Research

May 5, 2014

Presentation Outline

• The Momenta approach to product development

• Application to intravenous immunoglobulin (IVIg) and autoimmune diseases

• New products that may deliver enhanced benefit to IVIg

• Sialylation of IVIg

• Sialylation of recombinant IgG’s

• Recombinant products

2

Momenta Embraces ComplexityThree Critical Components to Deliver Medicines

Understanding the

nonlinear chemical

and biosynthetic

reactions that drive

production

Control of Manufacturing

*

*

High resolution physicochemical

analytics platform to fully

characterize any product

Thorough Structural

Characterization

13

Thorough Biological

Characterization

High resolution biology

analytics platform to fully

characterize any product

in non- and clinical

settings

• Generic Lovenox®• Generic Copaxone®

(under FDA review)

Biosimilars & Potentially Interchangeable Biologics

• M402 oncology Phase 1

Novel DrugsANDA Generics

Momenta has Extended this Approach to IVIgand Autoimmune Diseases

4

Complex Mixture Drugs

Intravenous Immunoglobulins

Complex Diseases

RA, Lupus, Psoriasis, etc

• IgG fraction derived from pooled plasma of ~10K donors

• Approved therapy for PID and 5 inflammatory diseases; used in ~100 other indications

• Poorly characterized mixture and MOA debated

• Dysregulation of complex immune system underlies >100 distinct diseases

• ~50M Americans affected

• Significant unmet needs despite recent advances

IVIg Is An Important….but Poorly Understood Anti-inflammatory Therapeutic

5

Widely Used as an Anti-inflammatory Poorly Understood MOA

Thorough Physicochemical Characterization Defines IVIg Composition

6

7

Thorough Physicochemical Characterization Defines IVIg Composition

100

200

300

400

500

21

>500

Number of distinct data readouts in characterization of IVIg lots

EU Pharmacopeia Momenta

IgG isotypesIgG1 Fc glycoformsIgG2 Fc glycoformsIgG3 Fc glycoformsIgG4 Fc glycoformsIgG2 disulfidesIgG4 mixed antibodiesIgG allotypesIgG Fc glycoform asymmetryConformational aggregatesAnti-idiotypic Ab complexesGlycationIgG1 Fab glycoformsIgG2 Fab glycoformsIgG3 Fab glycoformsIgG4 Fab glycoformsDeamidationOxidationGlycationDistribution of antigen specificityNon-IgG proteins……..

Dat

a R

ead

ou

ts

Thorough Biocharacterization Defines IVIgMechanism of Action

Genetic

Glycan

Protein and Cellular

Metabolic

8

Patients

Deep understanding of biological

mechanisms of action of existing

therapies & unmet medical needs

New Drugs

Combination of orthogonal analytics and informatics to deconstruct disease networks and complex mechanisms

Model Systems

CAIA

EAE

KBxN

ITP

Thorough Characterization Reveals New Products that May Deliver Improved Patient Benefits

• Sialylation of the Fc region of IVIg• Exploits natural mechanism of modulating IgG function

• Potential for lower dose, higher potency Ig products (e.g. SC delivery)

• Improved Ig products, targeted to existing or new inflammatory disorders

• Sialylation of the Fc region of biologic therapeutics• Enhanced anti-inflammatory activity of any recombinant biologic

• Novel Recombinant Products• Exploit major mechanisms of action of IVIg

• Homogeneous recombinant products with improved patient safety

• Potential to integrate anti-inflammatory properties of controlled sialylation

9

Alterations in the Sialylation of IgG are Associated with Autoimmune Disease and Disease Activity

10

Normal range

RA patients

Parekh et al., 1988

RA and JIA Patients have less Sialylation (and its Precursor Galactosylation) in their IgG

Vehicle

Fc

IVIG

sialylated Fc

K/BxN arthritis model

Anthony, R. & Ravetch, J et al., 2008US8470318US7846744

US 20130273040

sFc and Fc dose: 0.035 g/kgIVIG dose: 1 g/kg

Sialylation of the Fc region of IgG mediates enhanced anti-inflammatory activity

SCIENCE VOL 320 18 APRIL 2008

We Optimized the Enzymatic Process for Site-Specific Sialylation

11

+B4GalT1

ST6Gal10

20406080

100

Re

l. a

bu

nd

ance

(%

)

IgG1

IgG2

IgG3/4

020406080

100

Re

l. a

bu

nd

ance

(%

)

IgG1

IgG2

IgG3/4

0

10

20

30

40

Re

l. a

bu

nd

ance

(%

)

IgG1

IgG2

IgG3/4

> 80% Pure S2- IVIg

> 80% Pure S1-IVIg

IVIg Drug Substance

IVIg

S1-I

VIg

S2-I

VIg

12

Avoiding Undesirable Modifications Introduced in Original Process

ST6 enzyme produced from certain sources can introduce other chemical modifications into the product. These modifications could impact biological

functions, which could confound results if not controlled.

Modifications Introduced During Sialylation Process

ST6 Enzyme Source

Original Optimized

Carboxymethyl lysine 5% ND

Carboxymethyl arginine 10% ND

Methylglyoxal arginine 20% 2%

Imidazolidine 8% ND

Pentosidine Yes ND

Sialylated IVIg Suppresses Arthritis in the K/BxNSerum Transfer Model of Arthritis

13

Serum Arthritis31.01.2014 - 07.02.2014

0 1 2 3 4 5 6 7 8 9 100.0

2.5

5.0

7.5

10.0

12.5+1g/kg Mha-048674

+0.3g/kg Mha-048674

PBS

[days]

clinic

al score

treatment

IVIg 1 g/Kg

IVIg 0.3 g/Kg

PBS

Sialylated IVIg (S-IVIg) reveals better efficacy than IVIg. IVIg loses activity at 0.3 g/Kg whilst sialylated IVIg shows good activity at 0.3 g/Kg

Serum Arthritis31.01.2014 - 07.02.201414.02.2014 - 24.02.2014

0 1 2 3 4 5 6 7 8 9 100.0

2.5

5.0

7.5

10.0 +0.3g/kg JCl-044545

+0.1g/kg JCl-044545

+1g/kg Mha-048674treatment

PBS

[days]

clinic

al score

IVIg 1 g/Kg

S-IVIg 0.1 g/Kg

PBS

S-IVIg 0.3 g/Kg

In collaboration F. Nimmerjahn et al

Sialylation Is Required for Efficacy of IVIg in a Mouse Model of Immune Thrombocytopenia

0

5

10

15

20

25

30

35

G0

F

G1

F

G2

F

G0

G1 G2

G0

F+B

Glc

NA

c

G1

F+B

Glc

NA

c

G2

F+B

Glc

NA

c

A1

F1,3

A1

F1,6

A2

F

G1

F+N

eu

Ac

A1

1,3

A1

1,6 A2

A1

F+B

Glc

NA

c 1

,3

A1

F+B

Glc

NA

c 1

,6

A2

F+B

Glc

NA

c

G1

F+N

eu

Ac+

BG

lc…Re

lati

ve a

bu

nd

ance

(%

)

IgG1

IgG2

IgG3/4

IVIg

IgG1

IgG2/3

IgG4

0

5

10

15

20

25

30

35

40

G0

F

G1

F

G2

F

G0

G1 G2

G0

F+B

Glc

NA

c

G1

F+B

Glc

NA

c

G2

F+B

Glc

NA

c

A1

F1,3

A1

F1,6

A2

F

G1

F+N

eu

Ac

A1

1,3

A1

1,6 A2

A1

F+B

Glc

NA

c 1

,3

A1

F+B

Glc

NA

c 1

,6

A2

F+B

Glc

NA

c

G1

F+N

eu

Ac+

BG

lcN

Ac

Re

lati

ve a

bu

nd

ance

(%

)

IgG1

IgG2

IgG3/4

Desialylated IVIG

IgG1

IgG2/3

IgG4

14

Murine anti-CD41 Antibody-induced Thrombocytopenia Model

Increased Sialylation of IVIg Accelerates Platelet Recovery in a Murine Model of ITP

15

0102030405060708090

G0

F

G1

F

G2

F

G0

G1 G2

G0

F+B

Glc

NA

c

G1

F+B

Glc

NA

c

G2

F+B

Glc

NA

c

A1F

1,3

A1F

1,6

A2F

G1

F+N

euA

c

A1F

-Lac

NA

c

A1

1,3

A1

1,6 A2

A1F

+BG

lcN

Ac

1,3

A1F

+BG

lcN

Ac

1,6

A2F

+BG

lcN

Ac

G1

F+N

euA

c+B

Glc

NA

c

Re

l. a

bu

nd

ance

(%

)

IgG1

IgG2

IgG3/4

05

10152025303540

G0

F

G1

F

G2

F

G0

G1 G2

G0

F+B

Glc

NA

c

G1

F+B

Glc

NA

c

G2

F+B

Glc

NA

c

A1F

1,3

A1F

1,6

A2F

G1

F+N

euA

c

A1F

-Lac

NA

c

A1

1,3

A1

1,6 A2

A1F

+BG

lcN

Ac

1,3

A1F

+BG

lcN

Ac

1,6

A2F

+BG

lcN

Ac

G1

F+N

euA

c+B

Glc

NA

c

Re

l. a

bu

nd

ance

(%

)

IgG1

IgG2

IgG3/4

IVIg

sIVIg

Murine anti-CD41 Antibody-induced Thrombocytopenia Model

Sialylated IVIg Suppresses Skin Blistering in a Murine Model of Pemphigus

16

IVIg dose titration

EBA Score(14.02.2014 - 27.02.2014)

3 4 5 6 7 8 9 10 11 120

5

10

15

20

+0.3g/kg 048674

+0.6g/kg 048674

+1g/kg 048674

+Saline

days

aff

ecte

d b

ody a

rea [

%]

PBS

IVIg 1.0 g/Kg

IVIg 0.6 g/Kg

IVIg 0.3 g/Kg

PBS

IVIg 0.3 g/Kg

S-IVIg 0.3 g/Kg

3 4 5 6 7 8 9 10 11 12

0

5

10

15

20

days

aff

ecte

dbody

are

a[%

]

In collaboration F. Nimmerjahn et al

17

0

5

10

15

20 ****

day 1

2

aff

ecte

d b

od

y a

rea

[%

]

PBS

S-IVIg 0.3 g/Kg

S-IVIg 0.3 g/KgIVIg 0.3 g/Kg

Sialylated IVIg Suppresses Skin Blistering in a Murine Model of Pemphigus

In collaboration with F. Nimmerjahn et al

* p> 0.05*** p> 0.01

Thorough Characterization Reveals New Products that May Deliver Improved Patient Benefits

• Sialylation of the Fc region of IVIg• Exploits natural mechanism of modulating IgG function

• Potential for lower dose, higher potency Ig products (e.g. SC delivery)

• Improved Ig products, targeted to existing or new inflammatory disorders

• Sialylation of the Fc region of biologic therapeutics• Enhanced anti-inflammatory activity of any recombinant biologic

• Novel Recombinant Products• Exploit major mechanisms of action of IVIg

• Homogeneous recombinant products with improved patient safety

• Potential to integrate anti-inflammatory properties of controlled sialylation

18

Sialylation of Recombinant Fc or an Anti-Cytokine mAb Enhances Suppression of Murine Arthritis

19

Mea

n A

rth

riti

s Sc

ore Normal mice

Vehicle/DiseasePrednisoneIgG1 at 7.5 mg/KgSialylated IgG1 at 7.5 mg/Kg

MNT-KBN-004 Total Delta

0

20

40

60

IL-4, ic

sFc, 0.1 g/kg

Fc, 0.1 g/kg

PBS

IVIg, 1.2gm/kg

Ave

rag

e D

elt

a C

ali

pe

r M

ea

su

rem

en

t

Murine Anti-Collagen Antibody-Induced ArthritisMurine K/BxN Serum Transfer Arthritis

• Anti-inflammatory effect of Fc sialylation can be transferred to recombinant products

Thorough Characterization Reveals New Products that May Deliver Improved Patient Benefits

• Sialylation of the Fc region of IVIg• Exploits natural mechanism of modulating IgG function

• Potential for lower dose, higher potency Ig products (e.g. SC delivery)

• Improved Ig products, targeted to existing or new inflammatory disorders

• Sialylation of the Fc region of biologic therapeutics• Enhanced anti-inflammatory activity of any recombinant biologic

• Novel Recombinant Products• Exploit major mechanisms of action of IVIg

• Homogeneous recombinant products with improved patient safety

• Potential to integrate anti-inflammatory properties of controlled sialylation

20

21

Thorough Characterization of Kawasaki Disease (KD) Patients Reveals IVIg Treatment Signature

Kawasaki’s Disease Inflammation of the coronary

arteries pre-disposing to aneurysms (ballooning) and

stenosis (narrowing) of the arteries.

>700 analytes per sample

19,467 data points

In collaboration with J.C. Burns et al

Genome

Glycome

Proteome

Metabolome

Advanced Data Analytics Reveals IVIg Treatment Signatures

• Acute Phase

• FC-gamma Receptor Signaling

• Granulocyte Chemotaxis

• Defense Response to Bacterium

• Positive Regulation of NF-kappaB Activity

• ………..

22

Pri

nci

pal

Co

mp

on

ent

An

alys

isC

om

po

ne

nt

2

Component 1

Cluster 1 Cluster 2

Cluster 1: Active disease + developing aneurysms

Cluster 2: Resolved disease + IVIG response

Identified analytesand their associated pathways which are major differentiators

of cluster1 vs 2

P l a t e l e t L e v e l s D a y 5

I so

t yp

e (

1. 5

ug

) +

Sa

l in

e

CD

41

( 1. 5

ug

) +

Sa

l in

e

CD

41

( 1. 5

ug

) +

IV

I g a

t 1

g/ K

g

CD

41

( 1. 5

ug

) +

M- 1

13

at

0. 0

04

g/ K

g

CD

41

( 1. 5

ug

) +

M- 1

13

at

0. 0

2 g

/ Kg

CD

41

( 1. 5

ug

) +

M- 1

13

at

0. 1

g/ K

g

0

3 0 0

6 0 0

9 0 0n o r m a l r a n g e

Pla

te

le

t 1

09

/L

IVIg M-113Controls

Da

y 0

Da

y 1

Da

y 2

Da

y 3

Da

y 4

Da

y 5

Da

y 6

Da

y 7

Da

y 8

Da

y 9

Da

y 1

0

0

2

4

6

8

1 0

1 2

M - 1 1 3 , 0 . 1 g / k g

M - 1 1 2 , 0 . 1 g / k g

I V I g , 0 . 1 g / k g

P B S c o n t r o l

I V I g , 1 g / k g

Av

er

ag

e C

lin

ica

l S

co

re

Novel Recombinant Products Leverage IVIg MOA Resulting in Significantly Enhanced Potency

K/BxN Serum Transfer Arthritis Model

• >1,000-fold enhancement of molecular mechanism• >50-fold enhancement of in vivo efficacy versus IVIg

Murine anti-CD41 Antibody-induced Thrombocytopenia Model

Acknowledgements

• Jeffrey Ravetch, Rockerfeller University

• Falk Nimmerjahn, University of Erlangen-Nuremberg

• Robert Anthony, Harvard

• Jane Burns, UCSD

• Frank Austen, Harvard

• Momenta Team

24