AKTIV-DRY Meeting a Grand Challenge in Global Health Safer, Needle-Free, UnitDose, Inhalable Dry Powder Vaccines Scott Winston, Ph.D. Aktiv-Dry LLC AKTIV-DRY Inhalable Dry Powder Measles Vaccine Gates Grand Challenge in Global Health #3 Improve childhood vaccines: Develop needle-free delivery systems AKTIV-DRY Inhalable Dry Powder Measles Vaccine KEY PROJECT OBJECTIVES • Develop a stable respirable microparticulate dry powder measles vaccine by CAN-BD and manufacture under cGMP • Develop at least 2 simple, inexpensive, unidose Dry Powder Inhalers (DPI) that deliver well-dispersed fine particles to young children, adolescents, and adults • Assess drug product safety and immunogenicity in rodents and non-human primates, and file Investigational New Drug Application in India • Successfully complete a Phase I clinical trial of an inhalable dry powder measles vaccine in India AKTIV-DRY Inhalable Dry Powder Measles Vaccine TECHNICAL GRAND CHALLENGES • Maintain potency of attenuated live measles vaccine – Measles virus is inherently unstable – Most powder manufacturing processes use extreme conditions – Sorbitol is often used as a stabilizing sugar but may not be suitable for powder production – New excipient(s) may need to be identified as only two sugars, mannitol and lactose, are approved as excipients for inhaled drugs AKTIV-DRY Inhalable Dry Powder Measles Vaccine TECHNICAL GRAND CHALLENGES • Powder properties – Moisture content of powder must be very low (<1%) for stability – Particle size must be in the right range for inhalation (1-5 microns) – Powders must be free-flowing and easily dispersable – Powders must dissolve rapidly after reaching mucosal surfaces AKTIV-DRY Inhalable Dry Powder Measles Vaccine TECHNICAL GRAND CHALLENGES • Powder packaging – Develop container-closure system that is compatible with dry powder inhaler design – Container-closure system must be inexpensive – Packaging must be moisture resistant to maintain powder properties – Handling of container-closure materials and filling process must be scalable to potentially >100,000,000 doses AKTIV-DRY Inhalable Dry Powder Measles Vaccine TECHNICAL GRAND CHALLENGES • Dry Powder Inhaler – Inexpensive!! – Easy to use for mass vaccination campaigns – Easily transportable and disposable – Capable of dispersing powder agglomerates to generate high emitted doses with high fine particle fractions. – Consider more than one design to manage risk AKTIV-DRY Inhalable Dry Powder Measles Vaccine Commercial Rationale If vaccine is highly effective what is the commercial need? •  40% of current lyophilized vaccine is wasted each year. •  In 2007, 250,000 deaths from measles and 23 million children did not receive their primary immunization. AKTIV-DRY Inhalable Dry Powder Measles Vaccine Commercial Rationale •  No diluent is required •  Eliminates safety concerns with injections and contamination of multi-dose vials •  Single dose presentations are more costeffective • Improve the supply chain and transportation Nepal ©UNICEF PATH ©Dr. Gordon Perkin ©UNICEF Immunization in Difficult Circumstances 1999 Polio OutreachUpper Nile region of Southern Sudan AKTIV-DRY Inhalable Dry Powder Measles Vaccine Scientific Rationale If vaccination by injection is highly effective why deliver measles vaccine by inhalation? •  Inhalation mimics natural route of infection •  4,000,000 children in Mexico were successfully vaccinated with Wet Mist, an aerosolized EZ measles vaccine. • Measles antibody levels remain higher in children after vaccination by inhalation than with subcutaneous injection. Recombinant Measles Virus Expressing Enhanced Green Fluorescing Protein (EGFP) in the Lungs with Tracheo-Bronchial Lymph Nodes R. deSwart et al., PLoS Pathogen 3(11): e178 (2007) AKTIV-DRY Carbon Dioxide Assisted Nebulization with a Bubble Dryer® CAN-BD • In CAN-BD, the vaccine formulation solution/suspension in water is mixed intimately in a low volume tee with CO2 at 80-100 bar to form an emulsion. • The vaccine emulsion stabilized with myo-inositol is rapidly expanded to atmospheric pressure through a flow restrictor to generate aerosols of microbubbles and microdroplets. • The aerosol plume is dried at temperatures below 50-70 °C as it mixes with a sheath of nitrogen in the drying chamber. • Dry fine powders are collected on filters and packaged to exclude moisture (aluminum foil blister packs or capsules with Al-film/ polymer laminates for packaging). CAN-BD Nebulizer Near-critical or Supercritical CO2 (80 to 100 atm) Drug Solution or Vaccine Suspension Emulsion of product stream in liquid CO2 CO2 flashes upon rapid 100X decompression to atmospheric pressure 10 cm Flow Restrictor Tube ID = 75 µm Restrictor Tip AKTIV-DRY Dry Powder Measles Vaccine Manufacturing • Manufacturing and commercial partner is the Serum Institute of India (SII) – Located in Pune, close to Mumbai – Produces a wide-range of vaccines – World’s large manufacturer of measles vaccine-hundreds of millions of doses/year – Uses the Edmonston-Zagreb measles strain – WHO certified vaccine manufacturer and cGMP compliant GMP Dry Powder Measles Vaccine Production at Serum Institute of India AKTIV-DRY Myo-inositol Based Vaccine Formulations AKTIV-DRY Safety and Toxicology of Myo-inositol •  Myo-inositol is a naturally occurring compound found in large quantities in human plasma, tissue, and organs as well as many plants. •  Myo-inositol is on the GRAS list as a food substance and has been shown to be safe and well tolerated when given orally or intravenously up to 18 g/day. •  Published laboratory studies have shown myoinositol is NOT carcinogenic, mutagenic, or teratogenic. AKTIV-DRY Safety and Toxicology of Myo-inositol •  Aktiv-Dry conducted a GLP study in rats of myoinositol powder delivered by inhalation. •  Rats were exposed to a maximum of 30 mg/day for 3 consecutive. •  Endpoints include clinical pathology, histopathology, body and organ weights, clinical and cageside observations, and opthalmology •  No myo-inositol-related effects or delayed onset of toxicity was observed. AKTIV-DRY Myo-inositol Based Formulation of Dry Powder Measles Vaccine AKTIV-DRY Inhalable Dry Powder Measles Vaccine Dry Powder Characteristics Lot number SPC080115 Potency of bulk powder 441 PFU/mg (4.45 log CCID50/10mg) Stability Potency (after 7 days at 37 °C) 111 PFU/mg (4.07 log CCID50/10mg) (Passes WHO test for measles virus potency of less than 1 log loss) Potency of filled blisters 319 PFU/mg Particle Size Distribution FPF < 5.8 µm = 44.9% ± 3.6% FPF < 3.3 µm = 18.9 % ± 2.7% Moisture content 0.55% Bioburden Negative AKTIV-DRY Stability at 5ºC- Dry Powder Measles Vaccine AKTIV-DRY DRY POWDER INHALER DEVELOPMENT •  BD Technologies Solovent™ – Dry powder vaccine packaged in capsule with film seal on both ends – Energy source is syringe-pressure breaks film seal dispersing powder into spacer • Aktiv-Dry PuffHaler™ – Dry powder vaccine packaged in blister with removable lid – Energy source is squeeze bulb-pressure disperses powder from blister into reservoir Solovent™ Development Strategy Primary Container & Overwrap Package Energy Source Patient Interface Use Existing Solovent Platform & IP – Modular Components Configured To Commercial Powder Filling Systems Leverage Standard High Volume Production Methods Solovent™ Development Strategy Syringe Capsule Housing Film Capsule Film Spacer Adapter •  TOP-DOWN ASSEMBLY •  AXIAL DELIVERY FLOW PATH •  MINIMIZE SIZE & DEAD VOLUME •  CAPSULE FILMS PROTECTED DURING DEVICE HANDLING ADAPTIVE SPACER DESIGN FOR ADULTS & CHILDREN CONFIGURED AT TIME OF USE BD Solvent™ DPI Nasal Delivery of Fine Particles BD Solovent™ AKTIV-DRY PuffHaler® Aerosol Delivery System PuffHaler® Aerosol Delivery System •  Utilizes air expansion through a pressure release valve Side View Top View AKTIV-DRY Immunogenicity of Dry Powder Aerosol Measles Vaccine in Rhesus Macaques AKTIV-DRY Monkey Immunogenicity Study •  Collaborators Diane Griffin-Johns Hopkins University Paul Rota-CDC Mark Papania-CDC Steve Godin-Bridge Laboratories SII-BD-Aktiv-Dry-CU Project Teams Non-human primate measles model AKTIV-DRY Advantages •  “…best experimental model for studying pathogenesis of human measles” and “…premier model for final testing of candidate MV vaccines (Wyde et al., Vaccine, 2001). •  Shows human-like disease following exposure or innoculation with measles virus. •  Utilized in studies of aerosolized measles vaccine and dry powder measles vaccine (de Swart et al., Vaccine 2006 and 2007). •  Utilized in toxicology studies of aerosolized measles vaccine to support WHO/SII IND in India for Phase 1 clinical study. Non-human primate measles model AKTIV-DRY Disadvantages •  Monkeys are expensive; several thousand dollars each to purchase and maintenance costs are high. •  Handling is difficult and dangerous. •  Previous vaccine inhalation studies utilized anesthetization of animals which may have affected breathing patterns and tidal volume. •  Based on previous studies (de Swart et al., 2006 and 2007) viral replication and immune response to measles delivered by inhalation may be variable. •  Monkeys are a useful model but aren’t human. Rhesus Macaque Immunogenicity Study Key Objectives •  Demonstrate immunogenicity of measles dry powder vaccine in NHP. Measure both cellular and humoral responses. •  Deliver vaccine by inhalation with a mask using both PuffHaler and Solovent. •  Deliver vaccine by direct nasal delivery with a nasal prong using both PuffHaler and Solovent. •  Establish procedures for training macaques to accept aerosol vaccine delivery without anesthetizing the animals. •  Establish procedures to be used in NHP toxicology study. Assess general safety during the study. AKTIV-DRY Rhesus Macaque Immunogenicity Study AKTIV-DRY Protocol Design •  16 measles-seronegative rhesus macaques, juvenile or young adult, 2-5 kg in weight •  During quarantine period acclimate NHP to the procedures –  Acclimate NHP to restraining chair –  Train NHP to accept mask or prongs –  Train NHP to breathe placebo powders with masks or prongs •  Administer vaccine on SD1 •  Obtain bronchoalveolar lavage and tonsil swab samples on SD6 and ship frozen to CDC for PCR. •  Obtain blood samples on SD1, 14, 28, 56, 72, 98 •  Ship blood samples same day to JHU for T-cell assays and serum preparation. Ship frozen serum samples to CDC for ELISA and PRN assay. Monkey Immunogenicity Study Design Group Treatment Nominal Loaded Dose # of Animals 1 SII measles vaccine Subcutaneous 1000 PFU 2 2 SII measles vaccine Subcutaneous 100 PFU 2 3 PuffHaler-inhalation 50 mg powder 3086 PFU/4.72 log CCID50/10mg 3 4 PuffHaler-nasal 50 mg powder 3086 PFU/4.72 log CCID50/10mg 3 5 Solovent-inhalation 50 mg powder 4545 PFU/4.65 log CCID50/10mg 3 6 Solovent-nasal 50 mg powder 4545 PFU/4.65 log CCID50/10mg 3 AKTIV-DRY AKTIV-DRY Rhesus Macaque Immunogenicity Study General Observations During Vaccine Administration •  For a first-time use of PuffHaler and Solovent in NHP the study went remarkably well. •  Rhesus macaques could be trained to accept 5 administrations of vaccine with either PuffHaler or Solovent. Anesthesia not required. •  No noticeable changes in breathing patterns were observed during 30 seconds of inhaling powders through a mask. •  For nasal administration, PuffHaler seemed to be tolerated better than Solovent, possibly because of Solovent’s higher velocity of emitted dose and popping sound during actuation. Rhesus Macaque Immunogenicity Study AKTIV-DRY Viral Challenge Study •  An addendum to the study was approved by FNIH in July. •  All 16 monkeys will be transferred to JHU in September. Monthly samples for immunological assays will be taken until April. •  An additional 3, measles-seronegative, monkeys will be included to serve as challenge controls. •  All 19 monkeys will be challenged in April (1 year post vaccination) with the Bilthoven measles strain. •  Blood samples will be taken 2-3 times per week for 2 weeks, weekly for two weeks, then monthly for 3 months. •  Endpoints are viremia, rash, and immune response to challenge. AKTIV-DRY MONKEY IMMUNOGENICITY STUDY PCR RESULTS PAUL ROTA, CDC Group 1 1 Administration SC 1000 PFU SC 1000 PFU Sex Male Female Swab negative negative Copy # BAL Pellet negative negative Copy # 2 2 SC 100 PFU SC 100 PFU Male Female negative negative 3 3 3 PuffHaler Mask PuffHaler Mask PuffHaler Mask Male Male Female positive negative positive 18.30 positive positive 23.90 positive 4 4 4 PuffHaler Nasal PuffHaler Nasal PuffHaler Nasal Male Female Female positive positive positive 245.00 negative 85.70 positive 127.00 negative 5 5 5 Solovent Mask Solovent Mask Solovent Mask Male Male Female negative negative negative positive positive positive 1800.00 1110.00 488.00 6 6 6 Solovent Nasal Solovent Nasal Solovent Nasal Male Male Female negative negative negative positive positive positive 326.00 71.00 71.80 negative negative 678.00 549.00 5280.00 37.40 AKTIV-DRY Measles Neutralizing Antibodies (mIU/mL) in Serum Samples from Rhesus Macaques (Rota, CDC) mIU/mL 120 mIU/mL is considered protective AKTIV-DRY AKTIV-DRY SUMMARY AND CONCLUSIONS •  CAN-BD can be used to manufacture stable and potent dry powders of attenuated live measles vaccine with the necessary aerodynamic properties suitable for inhalation. •  Two low cost Dry Powder Inhalers, PuffHaler and Solovent, along with packaging configurations for dry powders, were developed. • Using both PuffHaler and Solovent deposition of measles vaccine dry powders into the lungs of cotton rats followed by viral replication was demonstrated. Induction of measlesspecific neutralizing antibodies was also observed. AKTIV-DRY SUMMARY AND CONCLUSIONS • Measles vaccine dry powders delivered by either PuffHaler or Solovent induced robust humoral and cellular immune responses in rhesus macaques. • High protective levels (>120 mIU/mL)of neutralizing antibodies were detected in all macaques immunized with measles vaccine dry powders. • Myo-inositol is a safe, non-toxic and stabilizing excipient in the measles vaccine dry powder formulation. AKTIV-DRY Inhalable Measles Vaccine Team Aktiv-Dry LLC, Prime Contractor; Bob Sievers, Principal Investigator; funded by a grant from the Foundation for the National Institutes of Health in the Grand Challenges in Global Health initiative. Subcontractors: Serum Institute of India BD Technologies University of Colorado National Jewish Medical and Research Center Johns Hopkins University University of Kansas RPC Formatec Consultants: John Carpenter, School of Pharmacy, CU Health Sciences Center Paul Rota and Mark Papania, US Centers for Disease Prevention and Control (CDC) University of Maryland Vaccine Research Center Winston Consulting Witham Consulting