Faster Vaccines for BiodefenseFebruary 18, 2010www.epivax.com

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•High throughput computing

•Immunoinformatics

•Vaccine design algorithms

•DNA synthesis, scale up, preparation of delivery device

•Delivery via iontophoresis / skin patch

•Animal safety/tox/immunogenicity/validation

•Deployment by mail/pharmacy distribution systems

Mandate – Faster, Safer Vaccines

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The mandate – Develop vaccines for WMD – pathogen unknown, immediately

Does this capacity exist? Yes.

The components:

This can be prebuilt!!

This can be prebuilt!!

Rapid deploymentwhen genome

sequence is in hand

Rapid deploymentwhen genome

sequence is in hand

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Infectious Disease-trained M.D. University of Chicago (1983),

Internal Medicine (NEMC 1986).

NIH NIAID / NCI: Additional training in

parasitology, immunoinformatics and vaccinology (1986-89).

CEO of EpiVax since May 1998.

$26M in NIH and foundation research funding.

Awarded $13M U19 award for vaccine design from the NIH in

July 2009.Associate Professor at

Brown University Medical School 1992- present;

Professor URI, Director I’Cubed, 2009 –present.

Says who? Annie De Groot M.D. CEO, EpiVax

EpiVax Management Teamhttp://www.epivax.com/team

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Dr. Anne De GrootCEO/CSO

Coordinates and directs strategy, business development and scientific programs.

William MartinCIO

Develops and sustains the infrastructure, informatics, business process, and IT systems in place at EpiVax.

Dr. Janet BuhlmannDirector of Molecular Immunology

Supervises and coordinates a portfolio of scientific projects involving Tregitope technology.

Dr. Leonard MoiseScientific Director of Vaccine Research

Directly responsible for scientific strategy and laboratory management of all biodefense and NTD vaccine programs.

EpiVax: Four Core Strengthshttp://www.epivax.com/services

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EpiVax Services Epitope Mapping

HLA Binding

T cell assays

In vivo assays (HLA Tg mice)

Fee for Service

EpiVax Vaccines

Grant funded R & D

Excellent proof of principle

Grant Funded(SBIR, R21, R01; >26M in funding since 1998)

EpiVax 2nd Generation Therapeutics

Select targets

Funded Res. Or Joint Devt

Develop molecule and license

Sponsored Research / Joint Development

Immuno-modulation

“Epi-13” Tregitopes

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

Options available for selected “Field of Use”

The Traditional Approach to VaccinesThe Traditional Approach to Vaccines

Effects of whole virus/bacteria unpredictable-contradictory;Effects of whole virus/bacteria unpredictable-contradictory;Process lengthy, prone to process issues, regulatory delaysProcess lengthy, prone to process issues, regulatory delays

Cross-Cross-reactive with reactive with Self or other Self or other PathogensPathogens

T reg T reg epitopes?epitopes?

Skew immune Skew immune response?response?

A new, evolved approach; GD-EDV

GD-EDV platformhttp://www.epivax.com/platform

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Genome-Derived, Epitope-Driven Vaccine Approach:

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In Silico EpiMatrix / ClustiMer / OptiMatrix [class I and class II alleles]

Conservatrix / BlastiMer/. EpiAssembler/ VaccineCAD

In Vitro HLA binding assay

ELISpot - ELISA - Multiplex ELISA - FACS - T regulatory T cell profiling

In Vector DNA prime/peptide (pseudoprotein boost) vaccines

Vaccine delivery / formulation optimization / detolerizing delivery agents

In Vivo HLA DR3, DR4 transgenic mice

HLA class I transgenic mice

Vaccination, Comparative studies

Emergency Use Authorization may obviate need for these

lengthy pre-clinical tests

Emergency Use Authorization may obviate need for these

lengthy pre-clinical tests

Proposed “Cassette” Approach to Faster Vaccines

“On Demand”“On Demand” PREPARED IN ADVANCE !PREPARED IN ADVANCE !

Step 1 Step 2 Step 3

Design can be done “on the fly”literally within 24 hours

What the proposed product? An ‘on demand’ vaccine In Silico – Discover Minimum Essential Units of Pathogens Driving Immune ResponseSafe: Eliminate all cross-reactive entities and toxicity in silicoConcatenate in “chain” of information using “VaccineCAD”Rapid insertion into DNA Plasmid; fast manufacturing scale up; Electroporation through skin (iontophoresis) on pre-manufactured micro needle patches

Components of supply chain that would be prepared in advance

Proposed Partners for Faster Vaccines Pilot Project

VennVax

Immunogenic

Epitopes

Shared

Immunogenic

Epitopes

SmallpoxVaccinia

GD-EDV Example - “VennVax”Smallpox Vaccine – Faster Safer Vaccine design

1. Downloaded Y16780(V. Minor), X69198 (V. Majo), L22579 Bangladesh and U94848 (Ankara), M35027 (Copenhagen, AF095689 (Xian tan), AY243312 (WR) Genomes from GenBankå

2. Identified highly conserved sequences - two methods (ICS and straight conservation)

3. 4. Identified potential Class II T cell clusters as well as A2, A24, B7, B44 epitopes

4. Analyzed Epitopes for homology to human sequences using BLAST Algorithm

5. Down selected best candidates

AF095689 (Xian tan)

M35027 (Copenhagen)

U94848 (Ankara)

L22579

L22579

Bangladesh

Bangladesh AY243312

AY243312(WR)(WR)

Y16780

Y16780

(V. M

inor)

(V. Minor)

X69198

X69198

(V. M

ajor)

(V. Major)

Genome-derived epitope driven vaccineConstruct Design / Assembly

DNA insertDNA insert

Intended Protein Product: Many epitopes strung together in a “String-of-Beads”Intended Protein Product: Many epitopes strung together in a “String-of-Beads”

Reverse Translation: Determines the DNA sequence necessary to code for the Reverse Translation: Determines the DNA sequence necessary to code for the intended protein. This DNA is assembled for insertion into an expression vector.intended protein. This DNA is assembled for insertion into an expression vector.

DNA DNA VectorVector

Protein product Protein product (folded)(folded)

Design the arrangement of the epitopes to minimize the immunogenicity of junctional Design the arrangement of the epitopes to minimize the immunogenicity of junctional

peptides and focus the immune response to the desired epitopespeptides and focus the immune response to the desired epitopes

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Improving Vaccine Design by aligning epitopes: Vaccine-CAD

De Groot AS, Marcon L, Bishop EA, Rivera D, Kutzler M, Weiner DB, Martin W. HIV vaccine development by computer assisted design: the GAIA vaccine. Vaccine. 2005

Example: Vaccine CAD

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Example: Vaccine CAD – final order

Result of Live Aerosol Challenge: 100% survival of GD-ED vaccinated mice vs. 17% of placebo

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0 20 40 60 80 100

100%

How to deliver Genome Derived –Epitope Driven Vax

DNA – chain of epitopes, or peptide in liposomes ICS-optimized proteins in VLPICS-optimized whole proteins

DNA Vaccines – A Faster SolutionRonald B Moss: Prospects for control of emerging infectious diseases with plasmid DNA vaccines

Journal of Immune Based Therapies and Vaccines 2009, 7:3

http://www.jibtherapies.com/content/7/1/3

Experiments almost 20 years ago demonstrated that injections of a sequence of DNA encoding part of a pathogen could stimulate immunity. It was soon realized that "DNA vaccination" had numerous potential advantages over conventional vaccine approaches including inherent safety and a more rapid production time. These and other attributes make DNA vaccines ideal for development against emerging pathogens. Recent advances in optimizing various aspects of DNA vaccination have accelerated this approach from concept to reality in contemporary human trials.

Although not yet licensed for human use, several DNA vaccines have now been approved for animal health indications. The rapid manufacturing capabilities of DNA vaccines may be particularly important for emerging infectious diseases including the current novel H1N1 Influenza A pandemic, where pre-existing immunity is limited. Because of recent advances in DNA vaccination, this approach has the potential to be a powerful new weapon in protecting against emerging and potentially pandemic human pathogens.

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If Needed - Validation: In Vivo Model for validation: HLA or humanized transgenic Mice

HLA DR3HLA A2/DR1

HLA DR1

HLA D2HLA A2 HLA B7

EpiVax Vaccine Program Overview

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• Smallpox – a safer VV vaccine?• “MDR” and “XDR” TB . . . (with Sequella)• Tularemia• Multipath (Burkholderia/Tularemia

. . . and with the DoD. . . A multi epitope vaccine for Ebola, EE’s, and Bacterial pathogens (Tularemia/Burk)

Current Collaboration with the DoD

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•EpiVax is currently funded to design, develop, and test a single T cell epitope based vaccine targeting three bacterial bioterror agents with NMRC (PI Dr. Andrea Keane Meyers). The targets of this program are Francisella tularensis (the causative agent of Tularemia and category A priority pathogen), Yersinia pestis (plague, a category A priority pathogen), and Burkholderia mallei (a category B priority pathogen). Funding for this project would enable and accelerate the EpiVax-NMRC vaccine design and development collaboration.• •EpiVax is also designing, developing, and testing a multipathogen vaccine targeting two viral bioterror agents with USAMRIID (PI Dr. Connie Schmaljohn). The targets of this program are Ebola (a category A priority pathogen) and Venezuelan Equine Encephalitis (VEE, a category B priority pathogen). Funding for this project would enable and accelerate the EpiVax-USAMRIID vaccine design and development collaboration.

•Both vaccines have dual military and civilian, war time and peacetime use. The goal of the combined program (USAMRIID and NMRC) is to develop a single, combined anti-bioterror vaccine. This Department of Defense-biotech collaborative project is consistent with the goals of the BDRD and DTRA chem-bio defense teams to develop vaccines against category A-C viral and bacterial bio-threat agents.

New tools for new vaccine challenges

• Characteristics of current vaccine development processes– Inefficient– Expensive– Time consuming (20+ years to vaccine)– High risk– Relatively unchanged for decades

• Vaccine Challenges and Emergent IDs - a new level of risk– Unprecedented infectious disease threats (population density, global travel, global warming,

widespread bioengineering technology and capabilities)– 10+ years from threat to vaccine is unacceptable– Advances in genomics, microbiology and immunology (including high throughput systems)

create an opportunity to re-think current processes

• Why wait? The time to begin “smarter, faster” vaccines is now!Is it time to Get “Smart” about Vaccines?

Who uses EpiVax Tools?

The U.S. Department of Defense*RocheAmgenEli LillyAbbott

Boehringer IngelheimBMS

And many, many more. . .

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Selected publications:• McMurry JA, et al. (TB) Current Molecular Medicine. 2007 Jun; 7(4): 351-68.• McMurry JA, et al. (Tularemia) Vaccine. 2007 Apr 20; 25(16): 3179-91. • De Groot AS, Moise L. (Review of tools). Exp. Rev Vaccines. 2007 Apr.: 6 (2): 125-7.• Knopf, P., et al. (C3d) Immunology and Cellular Biology. 2008 Mar-Apr;86(3):221-5.• L. Moise et al. (H. pylori). Human Vaccines 2008, Hum Vaccin. 2007 Dec 8;4(3).• A. S. De Groot, et al. (HIV) Identification of Immunogenic HLA-B7 “Achilles’ heel” Epitopes Within

Highly Conserved Regions of HIV. Vaccine. Vaccine. 2008 Jun 6;26(24):3059-71. • Gregory SH, Mott S, Phung J, Lee J, Moise L, McMurry JA, Martin W, De Groot AS.Epitope-based

vaccination against pneumonic tularemia. Vaccine. 2009;27(39):5299-306. • Leonard Moise, Julie A. McMurry, Soren Buus, Sharon Frey, William D. Martin and Anne S. De

Groot. In Silico-Accelerated Identification of Conserved and Immunogenic Variola/Vaccinia T-Cell Epitopes, Vaccine (David Weiner, special editor). http://dx.doi.org/10.1016/j.vaccine.2009.06.018 Vaccine. 2009 Jun 24.

• McMurry J, Johansson BE, and De Groot AS. A call to cellular & humoral arms: Enlisting cognate T-cell help to develop broad-spectrum vaccines against influenza A. Human Vaccine. 2008;4(2):148-57.

• De Groot AS, Ardito M, McClaine EM, Moise L, Martin W. Immunoinformatic comparison of T-cell epitopes contained in novel swine-origin influenza A (H1N1) virus with epitopes in 2008-09 Conventional Influenza Vaccine. Vaccine. 2009. Aug 3.

EpiVax: Four Core Strengths

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