DNA Vaccine for Chronic Hepatitis C



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Novel Vaccine Development (3:00PM~3:30PM) DNA Vaccine for Chronic Hepatitis C : Preclinical Evaluation of Immunogenicity and Safety of VGX-6150 Moonsup JEONG, Ph.D. Director of Pharma R&D Division GeneOne Life Science

Chronic Hepatitis C 130~170 million people are chronically infected with HCV (WHO statistics) - 2.2~3.0% of the world s population An estimated 3~4 million people are infected with HCV yearly (Bowen 2005, Thomson 2004, Levrero 2006) - 75~85% will develop chronic HCV infection - 60~70% will develop chronic liver disease - 5~20% will develop cirrhosis - 1~5% will die from cirrhosis or hepatocellular carcinoma *In total, 350,000 individuals die annually from HCV-related disease HCV infection is the leading cause for liver transplantation (CDC)

Medical Unmet needs Recent treatment (Direct-Acting Antiviral Drugs) - Improved SVR rates of 90% in treatment naïve patients Medical unmet needs - Unaffordable : High price ($1,000/dose for Sovaldi (Gilead), total $84,000) per patient - Unavailable : up to 75% of patients chronically infected with HCV are unaware of infection, precluding from treatment until diagnosed - Least effective in patients with advanced live disease - Associated with the development of viral resistance - Do not provide protection from re-infection Vaccine approach would be considered the most efficacious and cost-effective means

Ideal Vaccine for chronic hepatitis C Rationale for T cell vaccine to prevent and treat chronic hepatitis C - T cells play a critical role in viral control during primary infection - Keeping CD8+ T cells and CD4+ T cells continuously to remove persistent viremia viral eradication and effectively representing long-term clinical cure Effective vaccine against HCV - not need to provide sterilizing immunity - recapitulate and accelerate the immune pathway followed in natural infection - HCV T cell vaccine

DNA vaccine Definition Vaccines for protecting an organism against disease or curing disease by injecting it with genetically engineered naked DNA, plasmid, to produce an immunological response 5

DNA vaccine delivery by Electroporation 6

DNA vaccine as HCV T cell Vaccine Characteristics of DNA vaccine delivered by electroporation - Vaccine toxicity is virtually eliminated - Early and strong vaccine-specific immune response as evidenced by multiple non-human primate and clinical trials : Inducing robust, broad and poly-functional T cell immune responses - Stable at ambient temperature - Easy to manipulate - Quick and inexpensive production - Multiple immunizations to boost immune response without concerns of preexisting immunity DNA based immunization is an attractive vaccine modality for HCV

Antigen expressing plasmids Target antigens HCV Genome - Genotype : 1a/1b, 70% of all chronic infection - Type of antigens : 90% of non-structure proteins, 62% of HCV genome - Improving immune response for antigens Consensus sequence of antigen Engineering antigen gene : Codon/RNA optimization, IgE Leader sequence Electroporation delivery Modified pvax1 (Invitrogen)

Non-human primate Feasibility Study Immunization induces strong and broad HCV specific IFN response Rhesus macaques (Macaca mulatta) - Antigen specific IFN ELIspot Analysis - Strong and broad immunogenicity - Difference in immunodominant domains among outbred monkeys importance of broad immunity in individuals having genetic diversity

Non-human primate Feasibility Study HCV specific effective CD4+ and CD8+ T cells are elicited by immunization - Intercellular cytokine staining - Th1 or CTL cells were counted if produced one or more IFN, TNF, or IL-2 - Effective CD4+ and CD8+ T cells activation - Multi-antigenic vaccination is needed

Non-human primate Feasibility Study Vaccine-induced HCV specific cytolytic CTL remain detectable 6 weeks beyond immunization - Putative cytolytic CTL : positive staining for both surface CD107a and intercellular granzyme B (GrzB) - CD8+ T cell activation associated with acute resolution of HCV infection - GrzB production in response to antigen stimulation - Strong circulatory HCV-specific CTL

Non-human primate Feasibility Study Immunization gives rise to circulatory effector memory like CTL - Memory-like CD8+ T cells were characterized : CD54RA negative staining, CD27 staining - Effector Memory (EM: CD54RA-/CD27-), Central Memory (CM; CD45RA-/CD27+) - Phenotype similar to Memory T cells remained detectable in circulation of all monkeys

Non-human primate Feasibility Study Conclusions of monkey study - Immunization gave rise to IFN -secreting CD8+ T cells immunological determinant highly associated with acute HCV clearance - Upon recruitment of HCV-specific T cells to the liver late in acute infection, HCV replication is strongly inhibited due to IFN secretion effective means of controlling HCV replication w/o tissue damage - Significant frequencies of circulating cytolytic CTL, and significant increase of GrzB production within CD8+ population - HCV specific memory T cells have been shown to expand and could control secondary infection in human acute resolvers crucial for protection against reinfection Supporting clinical investigation of our HCV DNA vaccine

VGX-6150 Drug Product Composition Mixture of pgx8005, pgx8006, pgx8007 and pgx6005 with concentration of 10 mg/ml in 1 ml One vial contains Components pgx8005: plasmid DNA expressing HCV NS3/4A pgx8006: plasmid DNA expressing HCV NS4B pgx8007: plasmid DNA expressing HCV NS5A pgx6005: plasmid DNA expressing human IL-28B 3mg 3mg 3mg 1mg Formulation WFI (USP) 1mL

Immune-Adjuvant Target cytokine - IL-28B Type III Interferon (recently discovered, 2003) : IL-29 (IFN 1), IL-28A (IFN 2), IL-28B (IFN 3) Produced by DCs in response to viral proteins or Toll-like receptor agonist Same signaling pathways as type I IFNs to induce expression of IFN-responsive genes inhibiting replication of various viruses (HBV, HCV) - Immuno-Adjuvant function Typical T cell-stimulating better than cytokine IL-12 Good T cell benefit in antigen-specific IFN production Significant impact on long-lived antigen specific immune responses in CD8+ T cells (monkey study)

VGX-6150-01 clinical study synopsis Summary (ClinicalTrials.gov Identifier: NCT02027116) Title of study Principal Investigators Sponsor Multi-center, open-label, dose-escalation, phase I trial to evaluate the safety, tolerability and immunogenicity of VGX-6150 as second-line therapy in chronic hepatitis C patients Sang-Hoon Ahn, MD, PhD, Severance Hospital Jeong Heo, MD, PhD, Pusan National University Hospital GeneOne Life Science, Inc. Endpoints 1. Primary endpoints - Adverse Events (NCI-CTCAE 4.0, Lab test, Vital signs and ECG) - Pain during vaccination (VAS) 2. Secondary endpoints - Immunological response Antigen specific IFN-γ secreting T-cell response CD4+, CD8+ T-cell response, Antigen-specific IgG titer - Virologic response Serum HCV-RNA level

VGX-6150-01 clinical study synopsis Study flow per subject Dose escalation

VGX-6150-01 Follow-on clinical study synopsis Study flow per subject Overall schedule

Conclusions Our HCV DNA vaccine, VGX-6150, in non-primate study demonstrated strong immunogenicity, broad T cell reactivity, and T cell phenotypes closely associated with protection against chronic HCV infection VGX-6150 showed a favorable safety profile and was judged as well-tolerated VGX-6150 induced robust and strong T cell immune response dose-dependently IL-28B expressed from plasmid enhanced antigen specific T cell immune response more strongly as immune adjuvant We are now conducting phase I clinical study against chronic hepatitis C patients who failed SOC and DAA in Korea We are also preparing phase I clinical study in health adults as preventive vaccine against chronic hepatitis C in Korea

Acknowledgements Young K. Park, Byung Mun Cho, Hyojin Lee, Jooyeon Oh, Youngran Cho David B. Weiner Brain Latimer, Michele A Kutzler J. Joseph Kim, Niranjan Y. Sardesai, Mark Bagarazzi, Amir Khan, Matthew Morrow, Jian Yan, Kate Broderick, Xuefei Shen, John Stone Dorothy Peterson