Prevalence of HBV* by Region Epidemiology Prevalence of HBV* by Region Epidemiology Virology Vaccine Production Clinical Summary *Hepatitis B Virus

Worldwide HBV Infection Epidemiology Worldwide HBV Infection More than 2 billion people infected during lifetime Up to 2 million die each year from HBV infection Worldwide there are ~350-400 million HBsAg carriers In the WHO European Region, 14 million people are estimated to live with chronic hepatitis B; Up to 36,000 deaths are attributable to HBV every year in Europe Annual infection in US ~320,000, ~32,000 HBsAg carriers Persistent HBV is considered a significant risk factor for development of primary liver cancer Epidemiology Virology Vaccine Production Clinical Summary

Hepatitis B Virus Virology M H B s (Pre-S2) L S (Pre-S1) S S (S) c 3 . 2 k b D N A 4 n m Pre-S2 Epidemiology Virology Vaccine Production Clinical Summary

HBsAg – Protein Composition Virology HBsAg – Protein Composition Epidemiology Virology Vaccine Production Clinical Summary WHBV* rHBsAg (CHO-derived) *Wild type

Non-Response to Immunization Vaccines Factors Which Affect Non-Response to Immunization Epidemiology Virology Vaccine Production Clinical Summary enhancing Genetically determined resistance Advanced age Overweight Gender Smoking Chronic liver diseases Systemic diseases Immune suppression Pre-S attenuating Craven DE, et al. Ann Int Med1986; Alper CA, et al. N Eng J Med 1989, Milich DR Immunol Today 1988, Hohler T, et al. Hum Immunol 1998

Populations of Non-Responders to Conventional Vaccination Vaccines Populations of Non-Responders to Conventional Vaccination Epidemiology Virology Vaccine Production Clinical Summary Cancer patients (children) ~57% (31% on Rx 88% off Rx) Acute lymphocytic leukemia ~ 10% Bone marrow transplant patients 15-68% Chronic renal failure & dialysis 34-81% Patients with chronic liver disease ~50% Pre-transplantation candidates 28-36% Post-transplantation patients ~10% HIV (children & adolescents) ~30% Miscellaneous

Why Do We Need Better HBV Vaccines? Epidemiology Virology Vaccine Production Clinical Summary Non–response to conventional HBV vaccines in special populations Fast induction of immunity to HBV in defined populations Low compliance with the 3 dose regimen of conventional HBV vaccines Possible protection against HBV envelope mutant(s)

History of Hepatitis B Vaccines Epidemiology Virology Vaccine Production Clinical Summary 1st Generation: Plasma-derived -HBsAg, Pre-S 2nd Generation: Yeast-derived, (recombinant) -HBsAg, (S Antigen only) 3rd Generation: Mammalian cell derived (recombinant) -HBsAg, Pre-S2 -HBsAg, Pre-S2, Pre-S1

Three Generations of HB Vaccines Mammalian cell derived vaccines Epidemiology Virology Vaccine Production Clinical Summary Plasma derived vaccines rDNA Yeast derived rDNA Mammalian cell derived vaccines Sci-B-VacTM with the 3 epitopes

Third Generation Hepatitis B vaccine Production Sci- B-Vac™* Third Generation Hepatitis B vaccine Biosynthesized via recombinant DNA technology in engineered Chinese Hamster Ovary [CHO] cells harboring the entire HBs gene Manufactured under full GMP Produced in Chinese Hamster Ovary Cells and contains all three epitopes of the native virus surface antigen Epidemiology Virology Vaccine Production Clinical Summary *Previously manufactured under the trade names of Bio-Hep-B® and HepimmuneTM

Expression of the r-HBsAg in Sci-B-Vac™ Production Structure of HBV DNA Expression of the r-HBsAg in Sci-B-Vac™ Epidemiology Virology Vaccine Production Clinical Summary * Glycosylation sites (PreS2 Asn 4; S Asn 146)

Development of Recombinant HB Vaccine HBs Proteins (PreS1, PreS2, S) Production Development of Recombinant HB Vaccine + Epidemiology Virology Vaccine Production Clinical Summary Expression Vector (recombinant plasmid) HBs gene fragment rDNA Transfection Host (CHO cells) Expression HBs Proteins (PreS1, PreS2, S)

Development of Recombinant Production Development of Recombinant HB Vaccine Epidemiology Virology Vaccine Production Clinical Summary

Formalin Inactivation Release Criteria (Bulk) Tissue Culture Cell Propagation 1 Vial / WCB 3 – 9X10” cells Bioreactors Production Cell Inoculation Growth phase Production Phase Post Production Phase  Protein purification Purification Clarification Concentration Dialysis I DNasa treatment Dialysis II DEAE-I Anion Exchange Formalin Inactivation 0.2µ filtration bulk API Quality Control Release Criteria (Bulk) Identification Composition Sterility Impurities Endotoxin Protein Content DEAE-II Production Production of Sci-B-Vac ™ (Flow Chart 1) Epidemiology Virology Vaccine Production Clinical Summary

Release Criteria (Final) Production (Flow Chart 2) Epidemiology Virology Vaccine Production Clinical Summary Formulation Adsorption to Alum 20 µg/ml 10 µg/ml 5 µg/ml Final Filling Quality Control Release Criteria (Final) pH Alum content General Safety Potency Endotoxin Sterility Physical Inspection Volume in Container Visual Inspection 20µg/1ml 10µg/1ml 5µg/0.5ml 2.5µg/0.5ml

Surface Antigen Composition of Sci-B-Vac™ (SDS-PAGE & immunoblot) Production Surface Antigen Composition of Sci-B-Vac™ (SDS-PAGE & immunoblot) Epidemiology Virology Vaccine Production Clinical Summary ]S ]pre-S1 ] pre-S2

CHO Cells in Production Phase Epidemiology Virology Vaccine Production Clinical Summary

( Electron Microscopy ) Production HBsAg Particles ( Electron Microscopy ) Epidemiology Virology Vaccine Production Clinical Summary

Efficacy of Sci-B-Vac™ in Neonates (n=1206, Four Dose-Ranging Studies) Results of clinical trials Efficacy of Sci-B-Vac™ in Neonates (n=1206, Four Dose-Ranging Studies) Epidemiology Virology Vaccine Production Clinical Summary Seroprotection (%) Months P < 0.05 98 94 95 87 56 35 Injections: Study was conducted in endemic populations / Data on file

Efficacy of Sci-B-VacTM in Neonates (n=205, Comparative Study) Results of clinical trials Efficacy of Sci-B-VacTM in Neonates (n=205, Comparative Study) Epidemiology Virology Vaccine Production Clinical Summary GMT significantly* higher at all points in the Sci-B-Vac group 11 2 13055 6074 3211 722 Months GMT mIU/ml *p<0.01 Yerushalmi et al. – Ped. Inf. Dis. J 1997

Efficacy of Sci-B-Vac™ in Neonates (Born to HBsAg+ Mothers) Results of clinical trials Efficacy of Sci-B-Vac™ in Neonates (Born to HBsAg+ Mothers) Seroprotection (%) Sci-B-Vac = 5 g 20 50 Months 85 94 98 Epidemiology Virology Vaccine Production Clinical Summary Data on file

Efficacy of Sci-B-Vac™ in Children Results of clinical trials Efficacy of Sci-B-Vac™ in Children Epidemiology Virology Vaccine Production Clinical Summary Injections Age: 2m - 11y Anti-HBs, mIU/ml 1 10 100 1000 10000 100000 2 6 7 12 Months 2.5µg, n=33 5µg, n=36 Data on file

Results of clinical trials Immunogenicity of 2 Doses* Sci-B-VacTM in Adults (n=36, Comparative Study) 12 Injection 28,800 923 81 18.1 4.7 Epidemiology Virology Vaccine Production Clinical Summary Sci-B-Vac 10µg Engerix B 20µg Anti-HBs, mIU/ml *OFF LABEL Shapira et al. J. of Hepatology 2001

GMT Levels (mIU/ml) in Adults Results of clinical trials GMT Levels (mIU/ml) in Adults (n=476, Comparative Study) Months Anti-HBs mIU/ml Epidemiology Virology Vaccine Production Clinical Summary GMT values about 5 times higher at months 7 and 12 (P<0.0001) Raz R. et al. IMAJ 2001

Immunogenicity of a Sci-B-Vac™ According to Weight Results of clinical trials Immunogenicity of a Sci-B-Vac™ According to Weight Epidemiology Virology Vaccine Production Clinical Summary 10µg/ml Sci-B-Vac n=260 5µg/ml Sci-B-Vac n=253 20µg/ml Engerix B n=315 GMT mIU/ml (log) Data on file

Immunogenicity of Sci-B-Vac™ Results of clinical trials Immunogenicity of Sci-B-Vac™ in Dialysis Patients Epidemiology Virology Vaccine Production Clinical Summary Anti-HBs, mIU/ml 0 30 60 90 180 210 days 10000 1000 100 10 1 Sci-B-Vac – 20 g (n=9) HB-Vax- II – 40 g (n=12) Shouval D. et al. In: Viral Hepatitis and Liver Disease. Eds., K Nishioka, H Suzuki, S Mishiro, Toda, Springer Verlag, pp. 543,1994.

Results of clinical trials Efficacy of Sci-B-Vac™ in Non-and Low Responders to Standard Vaccine (N=719, Comparative Study) Epidemiology Virology Vaccine Production Clinical Summary Open, randomized, comparative, controlled, multi-center international parallel group study. To demonstrate superiority of Sci-B-Vac over standard vaccine after one or two additional doses in non-responders ≥4 prior injections of standard vaccine. To evaluate efficacy of the two vaccines in low responders after ≥4 and non responders after 3 prior injections of the standard vaccine. Rendi-Wagner P. et al. Vaccine 2006

Study Parameters Age: ≥18 years Randomized: 719 Results of clinical trials Study Parameters Epidemiology Virology Vaccine Production Clinical Summary Age: ≥18 years Randomized: 719 Injected with Sci-B-Vac: 479 Injected with Engerix B: 237 Injections: on days 1 and 85 Adverse Events: 3% Sci-B-Vac vs 0.8% Engerix B Rendi-Wagner P. et al. Vaccine 2006

Seroprotection After 1st or 2nd Additional Injection Results of clinical trials Seroprotection After 1st or 2nd Additional Injection (n=719, Comparative Study) Epidemiology Virology Vaccine Production Clinical Summary Seroprotection % Rendi-Wagner P. et al. Vaccine 2006

Analyses of 10,815 Individual Anti-HBs Measurements for 1,923 Vaccinees Epidemiology Virology Vaccine Production Clinical Summary The 3rd generation HBV vaccine Sci-B-Vac and HBV/MF59 appear the more immunogenic vaccines Hevac B and HB Vax vaccines appear less immunogenic Priming of immune memory is successful after a single dose Immune memory is maintained after anti-HBs antibody decay Antibody boosts following completion of conventional immunization are unnecessary Vaccines tested: Engerix B, SL*, Bio-Hep- B, China HBV MF/59, Recombivax, Herbiovac, Hevac B, HB Vax *Wilson J. et.al. 2002

Performance of Sci- B-Vac™ in > 20 Clinical Trials Epidemiology Virology Vaccine Production Clinical Summary Sci-B-Vac is safe and highly immunogenic Rapid (earlier/higher) immune responses High seroprotection rates Tested in adults and children in over 5,000 subjects Protects neonates of HBeAg positive and negative mothers Induces seroconversion in high-risk patients and in non-responders

Possible Future Applications For Sci-B-Vac™ Non / hypo responders to standard vaccines Vaccination of health care workers Immune suppressed patients Organ donors Universal vaccination for neonates in endemic countries using less than three doses Epidemiology Virology Vaccine Production Clinical Summary

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