EpiVax_Tregitope_Overview_2013

Tregitope Overview Non-confidential

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Annie De Groot M.D. CEO/CSO Bill Martin, COO/CIO

EpiVax Corporate Summary

• Founded 1998 – cash positive since inception

• Privately Held (De Groot, Martin Principals)

• Better Vaccine and Protein Design – Services

• Development of Immunomodulatory Products (Vaccines,

Tregitope)

• Successful mix of services and grant-funded research:

14% growth, year over year, without venture funding.

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Management Team http://www.epivax.com/about/management-team/

• Dr. Anne De Groot, CEO/CSO

• William Martin, Chief Information Officer

• Dr. Leslie Cousens, Director of Protein Therapeutics

• Dr. Lenny Moise, Director of Vaccine Research

• Anthony Marcello, Business Development Associate

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EpiVax: Four Core Strengths www.epivax.com

Epitope Services

Epitope Mapping

HLA Binding

T cell assays

In vivo assays (HLA Tg mice)

Fee for Service

Vaccines Grant funded R & D

Excellent proof of principle Grant Funded

2nd Generation Therapeutics

Select targets

Funded Res. Or Joint Devt

Develop molecule and license

Sponsored Research / Joint Development

Immuno-modulation

Tregitopes

In preclinical development- may be large market - allergy, autoimmunity

Options available for selected “Field of Use”

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Discovery of Tregitopes

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Tregitopes, discovered by Anne De Groot and Bill Martin at EpiVax, are linear

sequences of amino acids contained within the framework of monoclonal

antibodies and immunoglobulin G; the original finding was published in the journal

Blood in 2008. Since their discovery, EpiVax has compiled substantial evidence

linking Tregitopes to the activation of natural regulatory T cells. By selectively

activating these natural regulatory T cells, Tregitopes can dampen unwanted

immune responses.

Preliminary studies carried out by EpiVax and collaborators indicate that

Tregitopes may be useful for inducing tolerance to transplants, protein drugs, and

blood replacement therapies, as well as for the treatment of allergies. EpiVax

believes Tregitopes may explain the effectiveness of intravenous immunoglobulin

G (IVIG), widely utilized as an autoimmune treatment.

NIH SBIR grants coupled with private foundation funds have brought total pre-

clinical funding for Tregitopes to more than $3.4M in the past 4 years.

Tregitopes – Quick Summary


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EpiVax Screens mAbs

For immunogenicity

Found Epitopes that are

Highly conserved

Tolerated or ignored?

(David W. Scott connection)

Tregs?

. . . Test hypothesis

APC Tregitope peptide

derived from IgG


Published in Blood, 25 July 2008 Reprints available on request

Original Publication

http://www.epivax.com/publications/tregitope-publications/




Previous Evidence of Treg

Epitope in Fc domain

Constantin Baxevanis, et al.:

– 1986 hFc could induce tolerance in mice

– CH3 could not induce tolerance

– “Tolerogenic epitopes may exist in CH2 domain”

Eur. J. Immunol. 1986.16: 1013-1016

= No Tolerance

= Tolerance hFc

CH3

CH2

CH3

CH3

Fc Fusion Studies (David Scott Lab)

– 1996 Ag-IgG fusion could induce tolerance

– 2000 MHC CII essential in tolerance induction

– 2004 GAD-IgG induces CD4+CD25+ Tregs

–Also see work by Habib Zaghouani et al.

Evidence of Tregitopes in IgG

= Tolerance CH2

CH3

Ag

CH1

What’s the Evidence?

(1) HLA Binding

(4) Bystander Suppression

. . . and antigen-specific

tolerance induction

(2) nTreg induction

(3) aTreg induction

6/13/2013 10

(5) Modify APC Phenotype

Are there other examples in the literature?

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Edratide - TGF-beta and

FoxP3 expression (Human)

Edratide – HLA Restriction

Frame Frame DRB1*0101 DRB1*0301 DRB1*0401 DRB1*0701 DRB1*0801 DRB1*1101 DRB1*1301 DRB1*1501

Start Stop Z-Score Z-Score Z-Score Z-Score Z-Score Z-Score Z-Score Z-Score

1 GYYWSWIRQ 9 -1.01 -1.63 -1.79 0.16 0.07 -1.37 -0.74 -0.79 -0.42 0

2 YYWSWIRQP 10 -1.14 -0.15 0.31 0.37 1.11 -0.10 1.06 -0.48 -0.70 0

3 YWSWIRQPP 11 -1.18 0.14 0.52 0.68 0.89 0.80 1.16 0.31 0.40 0

4 WSWIRQPPG 12 -1.08 1.48 -0.31 1.05 0.05 1.19 1.77 0.14 0.15 1

5 SWIRQPPGK 13 -1.41 -0.08 -0.92 -0.50 -0.96 0.55 0.26 0.26 -0.01 0

6 WIRQPPGKG 14 -1.37 2.29 1.27 1.97 1.28 2.62 2.35 0.88 1.26 4

7 IRQPPGKGE 15 -1.66 0.12 0.10 0.08 0.18 0.63 -0.59 0.97 0.03 0

8 RQPPGKGEE 16 -2.54 -2.45 0.29 -2.38 -2.31 -0.21 -1.45 -0.13 -0.65 0

9 QPPGKGEEW 17 -2.14 -1.47 -1.99 -1.62 -1.00 -2.67 -2.50 -2.86 -2.21 0

10 PPGKGEEWI 18 -1.26 -2.63 -1.56 -3.15 -1.36 -1.52 -2.36 -1.54 -2.89 0

11 PGKGEEWIG 19 -1.12 -1.50 -1.15 -1.61 -1.87 -1.80 -0.76 -2.12 -0.89 0

DRB1*0101 DRB1*0301 DRB1*0401 DRB1*0701 DRB1*0801 DRB1*1101 DRB1*1301 DRB1*1501 Total

2.29 1.27 1.97 1.28 2.62 2.35 0.97 1.26 --

2.29 0.00 1.97 0.00 2.62 4.12 0.00 0.00 11.00

1.00 0.00 1.00 0.00 1.00 2.00 0.00 0.00 5.00

Top 10%* Top 5% Top 1%

Hydrophobic amino acid sequences scoring above 2.0 can be difficult to synthesize as peptides. Mutated amino acids are indicated in red typeface.

EpiMatrix Cluster Detail Report

File: MOZES_HCDR1 Sequence: HCRD1 Cluster: 1March 29, 2012 (Epx Ver. 1.2)

Z score indicates the potential of a 9-mer frame to bind to a given HLA allele; the strength of the score is indicated by the blue shading.

All scores in the Top 5% (Z-Score >= 1.64) are considered "Hits". *Scores in the top 10% (shown but not highlighted) are considered elevated, other scores are grayed out for simplicity.

Frames containing four or more alleles scoring above 1.64 are referred to as Epi-Bars and are highlighted in yellow. These frames have an increased likelihood of binding to HLA.

Frames conserved in IgG antibodies and believed to be either passively tolerated or actively regulatory are highlighted in green.

Flanking amino acids, added to stabilize the cluster during in-vitro testing, are presented in blue type face and underlined.

Total Assessments Performed: 88 Hydrophobicity: -1.09 EpiMatrix Score: 1.93 EpiMatrix Score (w/o flanks): 1.93

Scores Adjusted for Tregitope: -- EpiMatrix Score: -7.30 EpiMatrix Score (w/o flanks): -7.30

Hits

Summarized Results (29-MAR-2012)

Maximum Single Z score

Sum of Significant Z scores

Count of Significant Z Scores

AA Sequence &nbspHydro- &nbspphobicity

The hCDR1 peptide: GYYWSWIRQPPGKGEEWIG

The pCDR1 peptide: TGYYMQWVKQSPEKSLEWIG

The pCDR3 peptide: YYCARFLWEPYAMDYWGQGS

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DiaPep 277

Also somewhat HLA-restricted

Tregitope Collaberator Checklist - Relevant to Autoimmunity?

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Tregitopes induce adaptive tolerance in C57Bl/6, D011.10, OTII

Tregitopes suppress/treat diabetes in NOD model (Scott/EpiVax)

Tregitopes suppress transplant rejection in CD28 KO mice (Scott)

Tregitopes suppression = IVIG in OVA/Allergy Model (Mazer)

Tregitopes suppress immune responses to AAV capsid (Mingozzi)

Tregitopes cause expansion of Tregs – iTreg or nTreg?.

Details (and slides) for the above are available under CDA.

Contact: [email protected]

mailto:[email protected]



Anticipated Applications

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Antigen specific tolerance induction (De-immunization of biotherapeutics).

Treatment of Autoimmune Disease (MS, T1D, others yet to be

studied) Treatment of Allergy

How Tregitopes are used in our Web Based Immunogenicity Screening Platform - ISPRI

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Correlation of antibody immunogenicity without Tregitope adjusted EPX Scores

Correlation to observed Immunogenicity before accounting for Tregitopes

R2=0.17

Factoring in Tregitopes. . .

T cell response depends on:

T cell epitope content – Tregitope content + HLA of subject

Protein Immunogenicity can be Ranked

epitope

Protein Therapeutic

1 + 1 - Regulatory T cell epitope* = Response

epitope epitope

Immunogenicity Scale for Monoclonals

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6/13/2013 Confidential

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Correlation of antibody immunogenicity with Tregitope adjusted EPX Scores

Accounting for Tregitopes results in more accurate predictions.

Correlation to observed immunogenicity after accounting for Tregitopes

R2=0.76

Antibodies: A Special Case

- 80 -

- 70 -

- 60 -

- 50 -

- 40 -

- 30 -

- 20 -

- 10 -

- 00 -

- -10 -

- -20 -

- -30 -

- -40 -

- -50 -

- -60 -

- -70 -

- -80 -

IgG FC Region

Nuvion (0%)

Avastin (0%)

AB01 (EPX Adjusted Score: -46.98)

AB02 (EPX Adjusted Score: -44.48)AB03 (EPX Adjusted Score: -44.81)AB04 (EPX Adjusted Score: -45.81)AB05 (EPX Adjusted Score: -45.88)







Synagis (1%)

Simulect (1.4%)Humira (12%)

Bivatuzumab (6.7%)

Remicade (26%)

Rituxan (27%)

Campath (45%)

Humicade (7%)

Reopro (5.8%)

Tysabri (7%)

LeukArrest (0%)

Herceptin (0.1%)

New drug

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6/13/2013 Confidential

Due to the presence of Tregitopes, antibodies tend to fall lower

on the immunogenicity scale.

We have developed a refined method using regression

analysis to predict the immunogenicity of antibody sequences

based on observed clinical responses.

We have found that a balance in favor of Tregitope (regulatory)

content over neo-epitope (effector) content is correlated with

reduced clinical immunogenicity.

Ne

o E

pito

pe

Co

nte

nt

Tregitope Content

High Low Low

Avastin (0%)

Herceptin (0%)

Mylotarg (3%)

Simulect (1%)

Synagis (1%)

Hig

h

Campath (45%) Remicade (26%)

Rituxan (27%)

Leslie Cousens and Annie De Groot EpiVax, Inc.

Mechanism of Tregitope Action: In Vitro Studies

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APC

CD4

Teff

CD4+

CD25+

Treg Notch

MHC

Class

II

TCR

CD28/

CTLA4

CD80/

CD86

IL-10

Identifying Tregitope Mechanisms of

Action at the Immunological Synapse

ILT3

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Thanking our Collaborators

EpiVax, Inc. William Martin

Lenny Moise

Leslie Cousens

Tim Musset

Matt Ardito

Ryan Tassone

McGill

University Bruce Mazur

Ciro Piccirillo

Amir Massoud

Dartmouth

College Chris Bailey-Kellogg

Partial funding by

the NIH

Uniformed

Services

University of the

Health Sciences David Scott

Yan Su

Xin Li

University of

Maryland Achsah Keegan

Preeta Desgupta

Children’s

Hospital of

Philadelphia Katherine High

Federico Mingozzi

Daniel Hui

9/28/2012

Harvard Brigham &

Women’s Hospital Mo Sayegh

Nadar Najafian

Francesca D’Addio

Ciara Magee

Samia Khoury

Wassim Elyaman

Institute for

Immunology and

Informatics Anne De Groot

Loren Fast

Alan Rothman

Denice Spero

Duke University Priya Kishnani

Dwight Koeberl

EpiVax: Four Core Strengths

Confidential 25

EpiVax_Tregitope_Overview_2013

Business

z score

evidence of tregitopes

iggdiscovery of tregitopes

epivaxbelieves tregitopes

inducing tolerance

tregitopes quick summary

tolerance tolerogenic

epivax corporate summary