1. 1 Pandemic Influenza Vaccine Development Pandemic Influenza Vaccine Development sanofi pasteur R&D, France Frederick R. Vogel, Ph.D.

2. 2 Introduction Strategies for the Development of a Pandemic Influenza Vaccine Pandemic Influenza Preparedness sanofi pasteur France Preclinical Results (sanofi pasteur, Erasmus MC) Clinical Results (sanofi pasteur) sanofi pasteur United States Clinical Results (NIAID) Conclusions

3. 3 Strategies for the Development of a Pandemic Influenza Vaccine Strategies for the Development of a Pandemic Influenza Vaccine Short term (< 3 years): to develop as quickly as possible pandemic vaccines based on existing technology (Split vaccine/ egg-based technology) Medium / long term: to improve the performance of influenza vaccines and encourage R&D into new vaccine approaches, including cell culture technology

4. 4 Sanofi Pasteur’s Pandemic Preparedness: France Research program initiated in 2002 NIAID research grant: Production and testing of egg- and cell- based H5N1 and H7N1 vaccine strains FLUPAN (EC) contract: Cell-based H7N1 vaccine production in PER.C6  cells (Crucell NV) FLUPAN Phase 1 clinical study began in Norway in September 2006 Pilot scale egg-based production of H5N1 vaccines since 2004

5. 5 Preclinical: Immunogenicity and Protection from Challenge by an Alum-Adjuvanted H5N1 Vaccine in Cynomolgus Macaques C. Ruat 1 – C. Caillet 1 – J. Simon 3 – I. Legastelois1 – F. Pistoor 3 – R. Fouchier3 – A. Bidaut 2 - A. Osterhaus 3 1: sanofi pasteur2: sanofi-aventis3: Erasmus MC

6. 6 Immunogenicity Results: HI titers (chicken rbcs)

7. 7 Virology Results: PCR H5N1- specific TaqMan PCR in lungs H5N1- specific TaqMan PCR in pharyngeal swabs

8. 8 Summary - Preclinical An inactivated H5N1 pandemic flu vaccine, at a dose level of 30 µg HA adjuvanted with aluminum hydroxide (600 µg Al) was immunogenic in monkeys. Immunization with the alum-adjuvanted H5N1 did not induce disease exacerbation when monkeys were challenged with parental viral strain. Virus titration revealed that the animals receiving the aluminium hydroxide-adjuvanted vaccine were protected from viral challenge. Protection was also observed with the unadjuvanted vaccine.

9. 9 Clinical: H5N1 sanofi pasteur France Clinical: H5N1 sanofi pasteur France Design Randomised, open, multicenter study conducted in France 300 healthy adults, 50/group; 18-40 years old Six vaccine formulations: 7.5, 15 or 30 µg of hemagglutinin (HA) with or without aluminium hydroxide adjuvant (Ad) Two i.m. vaccinations (deltoid), 21 days apart Objectives Immunogenicity after each vaccination Safety profile: within 21 days following each injection Designed for EU Core Pandemic Dossier Lancet 2006. 367:1657-1664

10. 10 Assessments Safety D0-7 and D21-28: solicited AEs recorded Erythema, Swelling, Induration, Ecchymosis >0cm, Pain Fever (oral >37.5°C), Headache, Malaise, Myalgia, Shivering D0-42: SAEs and unsolicited AEs recorded Immunogenicity D0, 21, D42 assayed by UK HPA (M. Zambon) Haemagglutination inhibition (HI) using horse erythrocytes (LLOD 1:8) Seroneutralization (SN) assay (LLOD 1:2) Statistics Descriptive analysis on intent-to-treat population

11. 11 Clinical: Results Population 300 subjects completed up to D42 No drop-outs, lost to follow-up or withdrawals Mean age per group: 24 – 26 years Male/female per group: 0.8 - 1.9 Safety No SAEs D0-42 No fever with oral temp >38°C No severe injection site pain

12. 12 Immunogenicity Results Population Naïve pre-vaccination antibody below LLOD, given value of LLOD/2 (one positive subject by HI & SN, one borderline by SN only) Seroconversion=seroprotection Presentation of results Most relevant assessment criteria Distribution of titers i.e., various seroconversion thresholds Fold-rises

13. 13 Immunogenicity: Reverse cumulative HI titer distribution (horse erythrocytes) D42 D21 7.5µg 7.5µg+Al 15µg 15µg+Al 30 µg 30µg+Al

14. 14 Immunogenicity: Reverse cumulative Seroneutralization (SN) titer distribution 7.5µg 7.5µg+Al 15µg 15µg+Al 30 µg 30µg+Al D21 D42

15. 15 Clinical: Key Findings H5N1 vaccine is safe and well tolerated HI and SN results show similar trends Two doses needed to optimize immune response Two doses of 30µg+Al induced response in >60% of subjects: HI titer >32 & >2-fold rise in SN titer Encouraging immune response seen with lower dosages >40% of subjects seropositive (HI test) after two doses of: 7.5µg, 15µg or 15µg+Al Adjuvant effect is seen with 30µg HA after the 2nd dose

16. 16 Phase II safety and immunogenicity clinical study conducted in 2006 Core mock-up dossier preparation and potential rolling submission Next Steps: sanofi pasteur, France

17. 17 Sanofi Pasteur’s Pandemic Preparedness: United States In response to the US Government RFP May 2004: production of 8000 doses of a monovalent H5N1 vaccine for clinical trials sponsored by the NIAID September 2004: contract awarded for the supply of 2 million doses H5N1 vaccine from industrial scale batches November 2004: contract awarded to establish and maintain flocks of egg-laying hens for the manufacture of vaccine at full capacity on a year-round basis April 2005: contract awarded for the development of a cell culture vaccine September 2005: H5N1 vaccine stockpile Implementation of large scale production

18. 18 Sanofi Pasteur’s Pandemic Preparedness: U.S.A. A/Vietnam (H5N1) clinical doses (NIH/HHSN266200400031E)  Protective dose: 90 µg HA/1 ml dose Acquisition of H5N1 Vaccine (2005) Build pre-pandemic vaccine stockpile with current strain (HHS/HHSO100200500004C)   1.2m 90 µg doses for DoD   3000 30 µg/0.1 ml doses for NIH A/Mallard/Netherlands (H7N7) clinical doses (HHSO100200600023C) A/Indonesia (H5N1 clade 2) clinical doses (HHSO100200600023C) Acquisition of H5N1 Vaccine (2006 & 2007) Build pre-pandemic vaccine stockpile with current strain (HHS/HHSO100200700026I) Cell-based Preparedness Accelerate development of cell-based vaccine (CDC/HHS200-2005-11758) Egg-based Preparedness Annual clinical doses Year-round egg supply (CDC/200-2004-10431, HHSO100200600023C) High-Dose H5 clinical doses 30 µg/0.1 ml intradermal (HHSO100200500004C) Adjuvanted H5 clinical doses Dose-ranging study of alum adjuvanted vaccine (HHSO100200600021C mod 2) Development Stockpiling Contingency H5N1 Vaccine Services Build 2 million dose pre- pandemic stockpile (CDC/200-2004-09881, HHSO100200600021C)   90 µg HA/ml, 5 dose vial   NIH hyperimmune study   CDC employee protection 2004200520072006

19. 19 First clinical study (phase I) 451 healthy adults (18-64 years), 2 injections three weeks apart Dosage: 90, 45, 15, 7.5ug of hemagglutinin/placebo Conclusion: A two-dose regimen of 90 µg of subvirion H5N1 vaccine does not cause severe side effects and, in the majority of recipients, generates neutralizing antibody responses typically associated with protection against influenza N. Engl. J.Med. 2006. 354: 1343-1351 Second clinical study (phase II) Results of aluminum hydroxide adjuvanted H5N1 vaccine trial to be presented by NIAID at the WHO meeting in February 2007 H5N1 Clinical Results: sanofi pasteur US (NIAID)

20. 20 For all actors, including public health agencies and vaccine manufacturers, pandemic influenza vaccine development poses a challenge and requires strong cooperation Financial support by national authorities, permanent dialog and interaction between public health authorities and vaccine companies, associated with active collaboration with academic research institutions and biotech companies are needed to meet such challenge Conclusions