Transcutaneous Immunization Vaccines Delivered Via a Patch G Glenn

Innovative Administration Systems For Vaccines IOMAI Transcutaneous Immunization: Vaccines Delivered Via a Patch Gregory Glenn MD IOMAI IOMAI IOMAI IOMAI • Focus: delivery of vaccines to the skin • In Gaithersburg, 53 employees – Experienced management team – Preclinical, formulation, patch manufacture – Clinical testing, (2000 active patches placed) IOMAI IOMAI Vaccination via a Skin Patch • • • • • Scientific Rationale Human proof of concept Development strategy Patch technology Biodefense application IOMAI IOMAI Fundamental Observation • Adjuvants activate skin immune cells, enhance immune responses to vaccines • Potent adjuvants can be safely delivered via the skin • Antigens and adjuvants are readily delivered into the skin LT IOMAI Adjuvant IOMAI Transcutaneous Immunization: Antigen and Adjuvant Delivery via a Patch Langerhans cells Vaccine Patch Antigen + adjuvant Epidermis Migration Dermis Activated Langerhans cell w/ Flu antigen Draining Lymph Node Immune Response IOMAI Adjuvants Enhance Immunity in the Context of the Skin: Universal Finding 1.E+06 B IOMAI Influenza-Anti-NC IgG 1.E+05 36,034 Serum IgG Titer (EU) 25,845 1.E+04 1.E+03 1,433 1.E+02 9 ug HA no LT Wet patch 9 ug HA + 4.5 ug LT 9 ug HA (IM) IOMAI Transcutaneous Immunization: Delivery of Different Types of Vaccines and Antigens – Multivalent subunit vaccines • Trivalent split-virus influenza • ETEC colonization factors & toxin (CFA1, CS3, CS6, LT) • Conjugated multivalent polysaccharides, Hib • HIV • Anthrax (rPA) – Toxoid vaccines • Tetanus and diphtheria toxoids – Toxins - CT, LT, Exo A – Particles • Virosomal vaccines • Virus-like particles- human papilloma virus – Inactivated bacterial whole cells - H. pylori, ETEC – Live Viruses – DNA IOMAI IOMAI ESTABLISHED SCIENTIFIC PARADIGM Published by the American Society of Microbiology Infection and Immunity Transcutaneous Immunization with Bacterial ADP-Ribosylating Exotoxins, Subunits, and Unrelated Adjuvants Infect Immun 2000;68:5306 nature Skin immunization made possible by cholera toxin Nature 1998;391:651 AMERICAN SOCIETY FOR MICROBIOLOGY INTERNATIONAL WEEKLY JOURNAL OF SCIENCE Journal of Virology Transcutaneous immunization: T-cell responses and boosting of existing immunity Vaccine 2001;19:2701 accine V Immunostimulant patch containing heat labile enterotoxin from E. coli enhances immune responses to injeccted influenza vaccine through activation of skin dendritic cells. J Virol 2003;77:5218 advanced drug deliver reviews Transcutaneous immunization of domestic anim opportunities and challenges ADDR 2000;43:45 medicine nature Transcutaneous immunization: A human delivery strategy using a patch IOMAI Langerhans Cells • Network of barrier immune cells – Immune surveillance – Orchestrates immune response • • • • • High levels of co-stimulatory molecules/cytokine Antigen presentation results in T and B cell activation Cover 25% of human skin surface area, highly superficial Ideal targets for immune manipulation Activated by adjuvants delivered to the skin LCs in situ Human LC, 400x IOMAI IOMAI LT as Adjuvant • • • • • • • • LT ( heat labile enterotoxin from E. coli) Readily produced at commercial scale Natural product/natural human exposure Long history of experimental use as adjuvant Demonstrated clinical activity as adjuvant on the skin Potent Safely used on the skin-P2DBPC Stable/stable in formulation IOMAI Naive A 1 FITC B 38 FITC / LT IOMAI 70 C The Adjuvant LT Increases the Number and Activation State of Migrating LCs in the Lymph Node FITC D CD11c 55 E 60 F 76 MHC Class II G 38 H 22 I 67 CD86 Inguinal Cell Count Axillary K 3% J 62% Cervical L 0.6% IOMAI FITC IOMAI Clinical Proof of Principle • Safe • Adjuvant effect • Delivery of large proteins possible – Transcutaneous vs transdermal • Relevance of preclinical animal data? IOMAI Human Antibody Responses to Colonization Factor IOMAI (rCS6) and LT: Critical role of the adjuvant, delivery of large molecules 10000 A Anti-LT IgG 1000 B Anti-LT IgA 1000 100 100 10 10 1 LT + CS6 CS6 1 LT + CS6 CS6 1000 C Anti-CS6 IgG 1000 D Anti-CS6 IgA 100 100 10 10 1 LT+ CS6 CS6 1 LT + CS6 CS6 Vaccine group IOMAI Safety and immunogenicity of a prototype enterotoxigenic Escherichia coli vaccine administered transcutaneously. Guerena-Burgueno F, Hall ER, Taylor DN, Cassels FJ, Scott DA, Wolf MK, Roberts ZJ, Nesterova GV, Alving CR, Glenn GM. Infect Immun 2002 Apr;70(4):1874-8 IOMAI Gauze/Tegaderm “Wet” Patches 20000 Boost Boost Anti-LT IgG (ELISA Units) 15000 10000 Glenn et al Nature Medicine, Dec 2000 0 50 100 150 Days 200 250 300 5000 0 IOMAI IOMAI Conclusions • Large molecules readily delivered in context of human skin – LT 86kD – CS6 1,500 kD – Transdermals<0.5kD • Adjuvant plays critical role for immunogenicity • Responses robust-equivalent to natural immunity generated by ETEC infection IOMAI IOMAI Second Major Application • Transcutaneous immunization (TCI) – Adjuvant and antigen delivered together • Immunostimulant patch-(LT IS Patch) – LT alone in conjunction with injected vaccine IOMAI IOMAI IS Patch: Immunostimulation Using an Adjuvant Patch with Injected Vaccine Immunostimulant Patch LT Flu Vaccine Delivered by Needle Activated LCs Migrate To the Draining LN IOMAI IOMAI LT IS Patch for Influenza Split Virus Vaccine: Enhanced Immunnogenicity Two id Injections 5 ug Trivalent Flu Vaccine (Serum IgG to Johannesburg B Strain) 1,000,000 p = .0005 p = .0002 208,929 p = .0036 184,424 Serum IgG Titer (log) 93,585 100,000 10,000 8,868 1,000 no LT 5 ug LT 25 ug LT 50 ug LT LT patch Published in: Xabier et al, J. Virol, May 03 IOMAI Phase IIa LT IS Patch in the Elderly-Enhanced Immunogenicity Flu V002A Day 21 Seroconversion 100% 90% 80% IOMAI Young Elderly Eld/LT HAI geomean fold rise 70% 60% 50% 40% 30% 20% 10% 0% New Caledonia Panama Shangdong IOMAI IOMAI IOMAI IOMAI Making Products IOMAI LT Alone as Strategic Focus for Development of Skin Patches • Delivery,Safety, Stability • LT alone applications – IS Patch using LT alone with flu – LT ETEC immunogen, anti-toxin immunity IOMAI • Formulate manufacturable LT patches • Optimize using LT immune response readout • Leading to Patches with LT and 2nd antigen) – LT+Anthrax – LT+Flu –IOMAI Protein in Adhesive Patch IOMAI Backing Matrix with LT Release Liner IOMAI Patch Systems are: - Small and Thin - Comfortable To Wear - Readily Manufactured IOMAI IOMAI Protein in adhesive Patches Stabilize LT: Results for % Label Claim for PIA Patch 150 125 Upper Limit % Label Claim % Label Claim (Dose/Unit Area) 100 75 50 25 1 Yr Real Time Stability at 5C Degradation Studies Suggest Multi-year 5C Stability Potential for RT Stability Lower Limit % Label Claim -Dry -Stabilizing excipients -LT Robust 0 0 1 2 3 4 5 6 7 8 9 Time (Months) Patches were stored at 5 oC and sampled according a written stability protocol. SE-HPLC analysis was performed at the indicated time points. Patches were extracted with 200 mM NaPi for 1.5 hours. Supernatants were then analyzed for LT and aggregate content with G2000 SWxl size exclusion column on a Agilent 1100 HPLC System with a DAD detector set to 220 nm. The mobile phase was 200 mM NaPi with a flow rate of 1.0 mL/min. No notable degradation was detected at 8 months. IOMAI IOMAI IOMAI IOMAI Step 1-Hydration and SC Disruption Untreated SC Hydration Disrupted IOMAI IOMAI Effect of Human Stratum Corneum Disruption: Using LT Patches-anti-LT IgG as readout 90 LT IgG Fold Increase Over Day 0 80 70 60 50 40 30 20 10 0 50µg Hydration 50µg Tape-strip 50µg Pumice 50µg Emery Day 21 Day 42 400µg Hydration Figure 1: LT IgG response after one and two doses in NLT105 – Eight subjects per group were pretreated with the indicated methods as described, then vaccinated with 50µg or 400µg LT on Day 0 and Day 21. LT IgG ELISA was run on sera from Day 0, 21, and 42, reported as fold increase of relative ELISA units. From: Expert Review of Vaccines 2:253-267, April, 2003 IOMAI Consistent Responses to Single Application of LT in Protein in Adhesive Patches in the Elderly Surrogate for Delivery Anti-LT IgG IOMAI FLA102 45/Emery n= 8 Fold Rise 4.8 % seroconversion 88% FLA201a LT 45µg/Emery Placebo/Emery 100 100 4.9 1.0 76% 0% SLA103 45µg/Emery 45µg/Buffer/IPA 40 20 3.3 3.0 67% 70% IOMAI IOMAI IOMAI IOMAI Development Strategy • LT alone applications – Phase II evaluation • Superflu • ETEC Epidermis Immunostimulant Patch LT Flu Vaccine Delivered by Needle Flu V002A Day 21 Seroconversion 100% Vaccine Patch Langerhans cells HAI geomean fold rise 90% 80% 70% 60% 50% 40% 30% 20% 10% 0% Young Elderly Eld/LT LT+Ag Migration Draining Lymph Node Immune Response LT Specific ASC Responses New Caledonia Panama Shangdong Count / 10 cells 6 • TCI-adjuvant and antigen – Anthrax – Flupatch 140 Grp 4 IgG 120 100 80 Grp 5 IgG Grp 4 IgA Grp 5 IgA 60 40 20 0 Day 0 Day 9 Day 23 IOMAI TCI Anthrax Vaccine • • • • Spore forming, gram+ rod Inhalation anthrax 90% fatal Toxin-PA component binds PA antibodies protective IOMAI • • • • Anthrax vaccine by TCI rPA as antigen LT as adjuvant Delivered together IOMAI IOMAI Transcutaneous Immunization with rPA and LTAdjuvant Elicit Anthrax Toxin Neutralizing Antibodies B. anthracis toxin neutralizing antibody 30,000 25,000 20,000 15,000 10,000 5,000 0 LT (ug) rPA (ug) IOMAI Toxin Neutralizing Titer (ED50) Transcutaneous Immunization IM (rPA/alum) 20 20 4 20 2 20 0.4 20 0 20 20 0 10 WRAIR/IOMAI IOMAI Protection Against Inhalation Spore Challenge Group 1 2 3 4 5 6 7 No. of AJ mice 10 10 2 8 10 10 10 Vaccination (TCI) µ µ LT* (µg) rPA* (µg) 20 50 4 20 1 5 20 0 0 50 Alum 5 naïve Immunization Route TCI TCI TCI TCI TCI IM Challenge results Protection (%) 100 100 100 0 100 100 0 Immunized day 0, 14, and 28, challenged 3 wks after last dose IOMAI/DSTL IOMAI IOMAI Rabbits Immunized with a Protein in Adhesive Patch Containing LT and rPA 3 ds (d 0, 14, 28), TCI 2 wks post immunization Anti-rPA IgG 1.0E+06 A 1.0E+05 Serum IgG Titer (EU) Anthrax toxin neutralizing antibody 1,400 1,200 83,468 p = 8.4-e10 TNA Titer (ED50) 1,000 800 600 400 200 1.0E+04 1.0E+03 915 1.0E+02 1.0E+01 150 ug rPA 150 ug rPA 50 ug LT 0 placebo 150 ug rPA 150 ug rPA 50 ug LT Protocol 1198-107 IOMAI IOMAI Conclusions • Clinical experience in 16 trials validate the safety and immunogenicity of TCI • Clinical experience is critical for exposing the viability of vaccine technologies • Vaccine delivery via a patch is feasible and task is to develop a commercial product IOMAI IOMAI IOMAI IOMAI IOMAI CORPORATION IOMAI Selected Publications Needle-free skin patch vaccination method for anthrax, Matyas et al. Infect Immun (in press) IOMAI Transcutaneous immunization and immunostimulant strategies, Glenn et al. Immunol Allergy Clin N.A. (in press) Transcutaneous immunization and immunostimulant strategies: capitalizing on the immunocompetence of the skin, Glenn et al. Expert Rev Vaccines (2003);2:253 Immunostimulant patch containing heat labile enterotoxin from E. coli enhances immune responses to injected influenza vaccine through activation of skin dendritic cells, Guebre-Xabier et al. J Virol (2003);77:5218 Transcutaneous immunization, Glenn et al. New Generation Vaccines, 3rd Edition (Jan 2004) Safety and immunogenicity of a prototype enterotoxigenic Escherichia coli vaccine administered transcutaneously, Guerena-Burgueno et al. Infect Immun (2002);70:1874 Transcutaneous immunization using colonization factor and heat labile enterotoxin induces correlates of protective immunity for enterotoxigenic Escherichia coli, Yu et al. Infect Immun (2002);70:1056 Transcutaneous immunization: T cell responses and boosting of existing immunity, Hammond et al. Vaccine (2001);19:2701 Transcutaneous immunization with bacterial ADP-ribosylating exotoxins, subunits, and unrelated adjuvants.SchartonKersten Infect Immun (2000);68:5306 Transcutaneous immunization: a human vaccine delivery strategy using a patch, Glenn et al. Nat Med (2000):6:1403 Transcutaneous immunization: a new vaccine delivery strategy, Glenn et al. The Jordan Report 2000: Accelerated Development of Vaccines, 2000 Transcutaneous immunization with cholera toxin protects mice against lethal mucosal toxin challenge, Glenn et al. J Immunol (1998);161:3211 Skin immunization made possible by cholera toxin, Glenn et al. Nature (1998);391:851 IOMAI