Armed Forces Epidemiological Board

1. Armed Forces Epidemiological Board 20 May 2003 Medical Chemical Biological Defense Research Program U.S. Army Medical Research and Materiel Command LTC John P. Skvorak Director Medical Biological Defense Research Program U.S. Army Medical Research and Materiel Command Phone: 301-619-7889 john.skvorak@det.amedd.army.mil

2. Organizational Management & Execution • On 22 April 2003, USD(AT&L) approved the implementation plan for management of the DoD Chemical & Biological Defense Program (CBDP) • Management and integration of CB Defense Science & Technology (S&T) efforts assigned to the Defense Threat Reduction Agency (DTRA) • Establishment of a Joint Program Executive Office (JPEO) for the CBDP that reports through the Army Acquisition Executive (AAE) to the Defense Acquisition Executive (DAE)

3. Intelligence • Agent • Delivery System • Organization • Time Medical Countermeasures • Vaccines & Prophylaxes • Diagnostics • Therapeutics Chem/Bio Defense Doctrine Education & Training • Military and Civilian Health Care Providers • Electronic Communication • Distance Learning Physical Countermeasures • Detection • Physical Protection • Decontamination Protecting Warfighters Through Integration and Teamwork

4. Provide medical solutions for military requirements to protect and sustain the force in a Chemical and/or Biological Warfare environment To Preserve Total Warfighter Effectiveness on a CW/BW Battlefield Prevent casualties Provide effective treatment of casualties for rapid return to duty Provide rapid, far-forward diagnosis of CW/BW disease Medical Chemical and Biological Defense Research Program Mission & Vision

5. TEP, STARs, Regional , Proliferation, Technology, and other Assessments RDA and Modernization Plans, Budget Intelligence Requirements Process THREAT ASSESSMENTS • Prepared in discrete, tailored packages • Evaluate impact on users • Define mission needs REQUIREMENTS • Joint Requirements Office for Chemical, Biological, Radiological and Nuclear Defense (JRO-CBRN) • Joint Requirements Oversight Council (JROC) PROGRAMS • Joint Service Materiel Group (JSMG) • Defense Threat Reduction Agency (DTRA)/Joint Program Executive Office (JPEO) • OSD coordinates/integrates funding requests ICDs/CDDs/CPDs, CPR, Integrated Priority Lists All programs driven by validated threats and defined mission needs

6. Medical Biological Defense Business Area Taxonomy Medical Biological Defense Countermeasures (CMs) Diagnostics Bacteriology Virology Toxinology DTO CB.47 (ImmunoDx) DTO CB.31 (Brucella) Diagnostic Technologies DTO CB.24 (VEE) DTO CB.32 (Alt. Delivery) DTO CB.46 (Ricin) DTO CB.34 (Plague) DTO CB.54 (Pox Tx) DARPA Transition Vaccines Vaccines Vaccines Therapeutics Therapeutics Therapeutics Vaccines Therapeutics Genetically Engineered Threats Medical CMs Diagnostic Technologies

7. APG, MD Fort Detrick, MD Washington, D.C. MCBDRP Locations • Fort Detrick, MD • MCBDRP • U.S. Army Medical Research Institute of Infectious Diseases • Forest Glen Annex, MD • Walter Reed Army Institute of Research • Naval Medical Research Center • Washington D.C. • Armed Forces Institute of Pathology • Aberdeen Proving Ground, MD • U.S. Army Medical Research Institute of Chemical Defense • Natick, MA • U.S. Army Medical Research Institute of Environmental Medicine

8. Research Labs/CBMS Cooperative Management Research Laboratories BA3 BA4 BA1 BA5 BA2 Milestone A Milestone B MilestoneC Follow-on Production IPR Component Advanced Development IPR Basic Research Applied Research Production & Deployment Sustainment & Disposal Concept & Technology Development System Development & Demonstration PROGRAM Vaccine IPTOVERSIGHT SSC PSC PROGRAM EXECUTION Consolidated PDT Proof of Concept Baseline Product IdentifyThreatAgent Characterize ThreatAgent Identify Vaccine Antigens DefineAnimalModels Evaluate Vaccine Technical Approaches DevelopAssaysandReagents Define Manufacturing Process Phase 1 Clinical Trials FDA Licensure Pre-IND Activities IND Application to FDA ManufactureConsistency Lots Produce, Store andMaintainVaccineStockpile Post-MarketingSurveillance Formulate Multivalent Vaccine (if required) Phase 2a ClinicalTrials Design Surrogate End-point of Clinical Efficacy Phase 2b ClinicalTrials Validate Assays Qualify Assays SurrogateEfficacyTests Manufacture cGMP Pilot Lots Test in Animal Models Prepare andSubmit BLA Perform Non-clinical Tests Characterize Candidates Manufacture Small-Scale Pilot Lots Integration of DoD Acquisition and FDA Licensure

9. Medical Biological Defense Research Program

10. Bacteria: Bacillus anthracis (Anthrax) Yersinia pestis (Plague) Francisella Tularensis (Tularemia) Brucella sp. (Brucellosis) Burkholderia maellei (Glanders) Coxiella burnetii (Q Fever) Viruses: Smallpox Encephalomyelitis viruses Ebola Marburg Toxins: Botulinum Toxins (Types A – G) Staphylococcal Enterotoxins (SEA/B) Ricin Toxin Marine Neurotoxins Mycotoxins Clostridium perfringens Medical Biological Defense Potential Threats

11. Bacteriology Task Areas Objective:Explore the development of candidate prophylactic and therapeutic medical countermeasures (i.e., vaccines, antibiotics and biologics) against validated BW threat agents, using laboratory and appropriate animal models, and to demonstrate their capability for reducing mortality or incapacitation in animal models exposed to predicted or presumed battlefield doses of aerosolized bacterial BW agents. JFOC(s) addressed:Medical Prophylaxis (OC) – Individual Protection (PC) Medical Treatment (OC) – Restoration (PC) *Note: DTO CB.33 Recombinant Protective Antigen (rPA) Anthrax Vaccine Candidate completed in FY02

12. CB.33 Recombinant Protective Antigen (rPA) Anthrax Vaccine Candidate(DTO completed in FY02) • Requirement for transition of rPA vaccine candidate out of the tech base and into Technology Development pre-acquisition phase have been met • National Institute of Allergy and Infectious Diseases (NIAID) selected USAMRIID rPA candidate for Phase 1 clinical trials • IND submitted in 2QFY03 in support of Phase I and II clinical trials • Phase I trial to be initiated 2/3Q FY03 • USAMRIID rPA candidate and UK rPA candidate also part of long-term NIAID strategy for 25M dose stockpile Assembly and action of anthrax toxin

13. CB.31 Medical Countermeasures for Brucella(Continuing DTO) Brucellae in murine macrophage • Brucellosis • Nonspecific febrile illness • Low infectious dose, highly infectious by aerosol • Antibiotic resistance may limit treatment options • Vaccine Candidate • MNPH1 – live attenuated deletion mutant • Orally administered • Demonstrated proof of concept for protective efficacy in NHPs

14. CB.34 Recombinant Plague Vaccine(Continuing DTO) • Plague: Tech base research generated two recombinant vaccine candidates • Both rely on F1 and V antigens • U.S. candidate is a recombinantly produced fusion protein • UK candidate is a combination of the two individually produced proteins • Technical progress: • Demonstrated 80% efficacy in non-human primates against 2.8 x 104 colony forming units (CFU) • Transitioned F1-V vaccine candidate to technology development pre-acquisition phase

15. Non-DTO Supporting Efforts(Bacterial Vaccines) • Anthrax • Further define pathogenesis of anthrax infection • Evaluate other known antigens, new adjuvants, and delivery systems • Employ functional genomics effort to identify new virulence determinants • Plague • Characterize the host immune response • Plan and develop the next-generation plague vaccine through identification of new protective immunogens • Define the genetic diversity and pathogenesis of natural plague isolates

16. Non-DTO Supporting Efforts(Bacterial Vaccines) • Glanders and Melioidosis • Develop an animal model of aerosolized glanders infection • Determine the mechanisms of immunity and establish correlates of immunity • Identify new virulence determinants • Potential vaccine candidates for protection against glanders • Evaluate the basis for virulence of Burkholderia mallei and B.pseudomallei by genetic and molecular methods

17. Non-DTO Supporting Efforts(Bacterial Therapeutics) • Licensed antibiotics/novel antimicrobials • Primary focus is evaluation of licensed and investigational antibiotics for efficacy against BW bacterial threats • Specific effort to evaluate a series of licensed antibiotics as a post-exposure treatment for plague • Demonstrated proof-of-concept for small molecule inhibitors of anthrax toxin • Immunotherapy/Immunomodulators • Passive prophylactic antibody treatment protects animals against aerosolized plague or anthrax • Evaluating compounds capable of priming the immune response

18. Virology Task Areas Objective:Explore the development of candidate prophylactic and therapeutic medical countermeasures (i.e., vaccines and antivirals) against validated BW threat agents, using laboratory and appropriate animal models, and to demonstrate their capability for reducing mortality or incapacitation in animal models exposed to predicted or presumed battlefield doses of aerosolized viral BW agents. JFOC(s) addressed:Medical Prophylaxis (OC) – Individual Protection (PC) Medical Treatment (OC) – Restoration (PC) *Note: DTO CB.25 Multiagent Vaccines for Biological Threats completed in FY02

19. To move away from this… 25 Shots Plus Boosters To this… Marburg Virus Botulinum toxin Or this… Anthrax Marburg-Ci67 Marburg-Musoke Marburg-Ravn) Ebola-Z Ebola-S “Panfilovirus vaccine” CB.25 Multiagent Vaccines for Biological Threats(DTO completed in FY02) • A vaccine or delivery approach that can concurrently immunize an individual against a range of biological warfare threats. • Reduce vaccination schedules • Decrease production costs • Streamline regulatory requirements • RNA Replicon and DNA Vaccine

20. CB.24 Medical Countermeasures for Multivalent Equine Encephalitis Viruses(Continuing DTO) Live attenuated VEE vaccines derived by site-directed mutagenesis of a full length infectious cDNA clone Identification of attenuating mutations: • sequence of attenuated variants Wild-type full- pV3000 length cDNA clone mutagenesis of conserved sequences • pBR322 Site-directed mutagenesis e.g., Molecularly defined clone with attenuating mutations pV3526 RNA Testing in animal models Virus Rescue (V3526 virus) Multivalent VEE vaccine requirement can be met through the use of a single component vaccine (V3526), eliminating interference issues while greatly simplifying clinical trials and formulation issues

21. CB.54 Therapy for Smallpox and Other Pathogenic Orthopoxviruses(New DTO in FY03) • Cidofovir, sold under the trade name Vistide™, is approved by the FDA for the treatment of cytomegalovirus retinitis in AIDS patients • Cidofovir inhibits the viral DNA polymerase, thus stopping the replication of smallpox and other pathogenic orthopoxviruses and disease progression • Clinical trials of cidofovir against smallpox are not possible (smallpox has been eradicated) • FDA licensure is possible under the new Animal Efficacy Rule • Variola and monkeypox primate models are proposed to demonstrate antiviral efficacy

22. Non-DTO Supporting Efforts(Viral Vaccines) • Multivalent Equine Encephalitis (VEE/EEE/WEE) Vaccine (MEEV) • Use infectious cDNA clone & site-directed mutagenesis technology from DTO CB.24 to rationally derive live-attenuated eastern & western equine encephalitis (EEE & WEE) vaccine components to be combined with V3526 candidate into a MEEV • Investigate and assess alternative vaccine technologies • Recent Accomplishments • Developed an EEE vaccine component based on the cleavage deletion mutation • Successfully tested one WEE vaccine component in mouse model for: • Protective response within 30 days of vaccination • Duration of immunity of 1 year

23. Non-DTO Supporting Efforts(Viral Vaccines) • Filovirus Vaccine • Develop a multiagent vaccine capable of protecting against various Ebola (EBOV) and Marburg (MARV) viruses (panfilovirus vaccine) • Leverage vaccine platform technology from Multiagent Vaccine DTO (CB.25) & from outside collaboration • Next Generation Smallpox Vaccine • Leverage DNA vaccine approach from DTO CB.25 to identify orthopoxvirus immunogens that contribute to protective immunity • Test if a gene-based vaccine technology can serve as an alternative to the live-vaccinia virus vaccine, or as a prime to a vaccinia boost

24. FDA-Approved Compounds DARPA Academics FDA IND NCI-DTP library Commercial Sources Combinatorial Lib. Primates Mouse Model Commercial Sources Academics Cell-Based Assay In Vitro Evaluation Modeling Lead optimization Screening Lead Identification In Vitro Evaluation Assay Development Target Identification Target Validation Non-DTO Supporting Efforts(Viral Therapeutics)

25. Non-DTO Supporting Efforts(Viral Therapeutics) • Antivirals for Smallpox • Develop an oral therapeutic antiviral drug based on cidofovir to treat smallpox and other naturally occurring or genetically modified pathogenic orthopoxviruses • Develop a second therapeutic antiviral drug based on non-DNA polymerase target (recommendation from IOM) • Antivirals for Filoviruses • Identified filovirus targets • Develop high thorough-put screening assays • Identify lead compounds for filoviruses • Develop therapeutic antiviral drugs based on lead compounds

26. Non-DTO Supporting Efforts(Viral Therapeutics) • Immunotherapies for Filoviruses • Objectives • Develop immunotherapies for treatment for infection with the various filoviruses • Test therapeutic intervention in combination with immunotherapy • Technology • Murine monoclonals  humanized antibodies  production in plants (leverage ongoing DARPA transition effort) • Combinatorial library from survivor monkey/human  humanized antibodies • anti-EBOV equine IgG • Test intervention strategies to prevent and treat shock and hemorrhage

27. Toxinology Task Areas Objective:Explore the development of candidate prophylactic and therapeutic medical countermeasures (i.e., vaccines, pretreatments, drugs and immunotherapies) against validated BW threat agents, using laboratory and appropriate animal models, and to demonstrate their capability for reducing mortality or incapacitation in animal models exposed to predicted or presumed battlefield doses of aerosolized toxin BW agents. JFOC(s) addressed:Medical Prophylaxis (OC) – Individual Protection (PC) Medical Treatment (OC) – Restoration (PC)

28. SoloVentTM MicromedicaTM micro-needles OnVaxTM “swipe and go” V. J. Sullivan, Ph.D. Copyright BDT 2000 CB.32 Alternative Vaccine Delivery Methods(Continuing DTO) BD Technologies Proprietary Alternate Vaccine Delivery Devices Currently Under Evaluation • Respiratory, transdermal, oral immunization that is safe, efficacious & expedient for stimulating mucosal and systemic immunity. • Simplify administration of the multiple vaccines. • Evaluation of multiple novel adjuvants in combination with alternate deliveries

29. CB.46 Recombinant Ricin Vaccine(New DTO in FY03) Ricin: Classical approaches to producing a ricin toxoid vaccine proved unsuitable • Recombinant expression vectors will be used to produce mutated A-chain immunogens capable of protecting against ricin toxicity • Technical progress: • Designed, expressed, purified and partly characterized ricin vacine candidates • Lead candidate lacks enzyme activity and protects mice against ricin lethality • Lead candidate is stable, monodisperse monomer under physiological conditions with a reduced tendency to self-aggregate

30. Non-DTO Supporting Efforts(Toxin Vaccines) • Toxin vaccines • Recombinant candidates for BoNT serotypes A, B, C1, E, and F (previous DTO) • Candidates transitioned to advanced development • Effort limited to support of advanced developer • Staphylococcal enterotoxin (SE) A and B vaccines (previous DTO) • Candidates ready to transition • Effort limited to stability analysis on pilot lots for use in future clinical studies

31. POST-EXPOSURE TREATMENTS (5-10 years out) PRETREATMENTS (5 years out) Passive Immuno- therapy Receptor Antagonists Scavenge Free Toxin Active-site Inhibitors Protect SNAREs Rescue SNAREs NEURON DRUG DELIVERY (10-15 years out) Bot Hc Delivery Vehicle Streptolysin-O Non-DTO Supporting Efforts(Toxin Therapeutics) • Toxin therapeutics • Therapeutics for botulinum neurotoxins • Current major efforts • Active site inhibitors • Immunotherapy (MAbs) • Expand structural biology and high performance computing capabilities and access to compound libraries

32. Non-DTO Supporting Efforts(Toxin Therapeutics) • Therapeutics for exposure to SEs • Block T-cell activation • Block SE-stimulated cytokine release • Therapeutics for exposure to Ricin • Basic research effort

33. Diagnostic Technologies Task Area Objective:Explore the development of technology candidates (reagents, protocols and devices) for inclusion into a deployable state-of-the-art identification and diagnostic system that integrates multiple methods for the identification of potential BW agents and the diagnosis of diseases they cause. The technology aim is to develop and integrate technologies so they will be capable of identifying multiple independent biomarkers from different agents simultaneously. JFOC(s) addressed:Medical Diagnostics (OC) – Restoration (PC) Med and Env Surveillance (OC) – Contamination Avoidance (PC) *Note: DTO CB.26 Common Diagnostic Systems completed in FY02

34. CB.26 Common Diagnostic Systems(DTO completed in FY02) Common Diagnostic Systems Delivered by S&T Program • Milestone A in FY01 • Technology Data Package on evaluation of diagnostic technologies in support of JBAIDS • Continuing to transition reagents and assays to the Critical Reagents Program • Provides capability to rapidly identify exposure to BW and infectious disease agents • Portable nucleic acid analysis system: • Specimen processing, gene detection device, reagents, protocols • Capable of simultaneous detection of multiple agents

35. CB.47 Improved Immunodiagnostic Platform(New DTO in FY03) Technology Options • Adjunct to nucleic acid detection • Primarily for the detection and identification of toxin threats • Also provides confirmatory assay for other medical diagnostic tests Time-Resolved Fluorescence Electrochemiluminescence (ECL) Reaction - First Generation Device - ORIGEN® Magnetic Field Detection Luminex

36. Non-DTO Supporting Efforts(Diagnostics Technologies) Evolutionary Strategy • Strategic Plan • Assay development • Identification of novel biological targets • Testing and evaluation • Technology assessment 2002 Portable Rapid Nucleic Acid Analysis System (Live Agents) Block I (CB.26) Improved Immunodiagnostic System (Toxins) Block II (CB.47) 2004 Integrated Diagnostic Systems Block III 2007+

37. Genetically Engineered Threats Countermeasures Task Area Objective:Identify, group, prioritize, and assess the medical impact of non-traditional toxins, virulence factors, and genetically engineered microbes as BW threat agents. The information generated from research efforts under the thrust area will be used to direct the development of medical countermeasures and diagnostics against the most plausible and dangerous novel or emerging BW threats. JFOC(s) addressed:Medical Prophylaxis (OC) – Individual Protection (PC) Medical Treatment (OC) – Restoration (PC) Medical Diagnostics (OC) – Restoration (PC)

38. DARPA Transition of BW Defense Programs Thrust Area • Objective:Pursue technological advancement of novel medical countermeasures and diagnostic technologies initially developed through the Defense Advanced Research Projects Agency (DARPA) that will protect the warfighter from BW agents and provide increased operational capabilities. • Specifically, these promising initiatives will be developed to demonstrate proof-of-concept in appropriate animal models and include: novel, broad-spectrum therapeutics; the application of gene shuffling technology to DNA vaccine development; plant-based production of vaccine antigens and antibodies by expressing plague antigens and Ebola monoclonal antibodies in transformed plants; and a hand-held, tissue based biosensor through step-wise development of B-cell lines for relevant pathogens and multi-channel sensors. • JFOC(s) addressed:Medical Prophylaxis (OC) – Individual Protection (PC) • Medical Treatment (OC) – Restoration (PC) • Medical Diagnostics (OC) – Restoration (PC) • Status: Nine programs selected to date

39. CB.34 Plague Licensed Plague Plague Vaccine Licensed NGAV Roadmap(Vaccines) 98 99 00 01 02 03 04 05 06 07 08 09 10 11 12 13 Vaccine Technologies S&T Program CB.25 Multiagent CB.32 Needle-less Delivery CB.31 Brucella Brucella Vaccine CB.24 Multivalent VEE Multivalent VEE Vaccine Multivalent VEE Vaccine CB.33 rPA Next Gen Anthrax Vaccine (NGAV) CB.46 Ricin Ricin Vaccine FDA Licensed Vaccine Procurement Tech Base VEE – Venezuelan Equine Encephalitis rPA – recombinant Protective Antigen Developmental Un-funded Developmental Partially funded Developmental

40. Roadmap(Diagnostics and Therapeutics) 98 99 00 01 02 03 04 05 06 07 08 09 10 11 12 13 Diagnostic Technologies S&T Program CB.26 Common Diagnostic System JBAIDS Blk I CB.47 Improved Immuno-diagnostic Platform JBAIDS Blk 2&3 JBAIDS Blk 2&3 Therapeutic Technologies S&T Program Therapy for Smallpox & other Pathogenic Orthopoxviruses CB.54 Therapy for Smallpox & other Pathogenic Orthopoxviruses FDA Approval of Cipro® for use after exposure to inhalation anthrax (Aug 2000) Tech Base Procurement JBAIDS – Joint Bio Agent Identification & Diagnostic System CDS – Common Diagnostic Systems for Bio Threats & Endemic Infectious Diseases Developmental Un-funded Developmental Partially funded Developmental Unfunded Procurement

41. FY03 Congressional Interest(Medical S&T Funding Increases: +$20.4M) • Engineered Pathogen Identification and Countermeasures Program (“Bug to Drug”) (Sarnoff Corporation): +$5.0M • Monoclonal Antibody-based Technology “Heteropolymer” (HP) System (EluSys Therapeutics, Inc.): +$1.0M • Bioadhesion Research (Ligocyte Pharmaceuticals, Inc.): +$1.8M • Mustard Gas Antidote (Mustard Consortium): +$2.1M • Needleless Delivery Methods for Recombinant Protein Vaccines (BD Technologies): +$1.0M • Vaccine Stabilization (U of Kansas): +$1.5M • Organic Vaccine Production (TBD): +$2.5M • Portable Biochip Analysis System (Cleveland Clinic): +$1.8M • Piezoelectric Dry Powder Inhalation Device (U. of Pitt) : +$1.7M • Bioprocessing Initiative (U. of Nebraska Lincoln): +$2.0

42. Cooperation with the Department of Health and Human Services (DHHS) • NIAID/CDC/FDA anthrax therapeutics • NIAID/USAMRIID/JVAP rPA clinical trials • NIAID/USAMRMC/USAMRIID Biodefense Campus • Program coordination • Program management • Infrastructure • USAMRIID/CDC • Smallpox research program • Anthrax antibodies

43. Medical Biological Defense Future Trends • Countermeasures for Genetically Engineered Microbes • Genomic sequencing of BW threat agents to identify and understand virulence factors, toxins and drug resistance genes • Immunomodulators and Therapies • Alternatives to agent-specific vaccines or therapies • Multiagent Vaccines • Alternative to one vaccine for one BW threat agent • Alternative vaccine delivery strategies • Immunization via mucosal and transdermal routes • Early markers of infection/host response