Inhalable Dry Powder Aerosol Attenuated Live Virus Measles Vaccine R. Sievers 1,3 , S. Winston 1 , D. Griffin, P. Rota, B. Quinn 1 , J. Searles 1 , D. Krank 1 , P. Bhagwat 1 , P. Pathak 1 , L. Rebits 1 , R. Dhere 2 , V. Vaidya 2 , R. Muley 2 , J. Burger 3 , D. McAdams 3 , S. Cape 1,3 K.Powell 4 , C. Shermer 4 , L. Chan 4 and K. Kisich 5 1 Aktiv-Dry LLC, 6060 Spine Rd., Boulder, CO USA, 2 Serum Institute of India Ltd., Pune, Maharashtra, India, 3 University of Colorado, Boulder, CO USA 4 BD Technologies, Research Triangle Park, NC, USA 5 National Jewish Medical and Research Center, Denver, CO USA
Well-formulated free flowing stable microparticles with less than 1% residual moisture, with high virus titers, rapidly dissolved, to replicate and create an immune response.
Inexpensive unidose blister packs or capsules with overwraps to protect vaccine from contamination, decomposing, reaction with water, oxidants and UV, and to reduce vaccine wastage. (Presently 40% is destroyed.)
Simple active dry powder inhalers that can disperse powder agglomerates to generate high emitted doses with high fine particle fractions.
ADVANTAGES OF AEROSOL DRY POWDER OVER LIQUID VACCINES
Powders inherently more stable than liquids
No line current or batteries required for aerosolization
Needle-free vaccination by inhalation of an aerosol of fine dry powder microparticles,~1-5 microns MMAD
In-vivo rapid dissolution in moist respiratory mucosa
Replication of live-attenuated measles vaccine virus
Robust immune response demonstrated by plaque neutralization in two animal models: Rhesus macaques and Cotton rats; macaques will be challenged with wild-strain of measles virus one year after immunization.
After demonstration of inhalation safety of the myo-inositol stabilizer in Sprague-Dawley rats and the E-Z live-virus vaccine in Rhesus macaques, an IND will be submitted for Phase I clinical trials in India.
Particles formed at 50 - 60 °C from an aqueous solution containing 11% total dissolved solids (50 g/L myo-inositol, 25 g/L gelatin, 16 g/L arginine-HCl, 1 g/L alanine, 2.1 g/L histidine, 3.5 g/L lactalbumin hydrolysate, 3 g/L tricine, pH 6.5 - 7.0). Microparticles averaged 1 µm in diameter and encased spherical virus particles averaged 120 nm.
Mean viral potency of measles vaccine powder samples was 4.6 log CCID50 / 10 mg.
FREEZE-FRACTURE SEM OF E-Z SUB-MICRON MEASLES VIRUS ENCASED IN MYO-INOSITOL-STABILIZED EXCIPIENT MICROPARTICLES
SAFETY OF MYO-INOSITOL STABILIZER AND REFORMULATED E-Z LIVE-VIRUS MEASLES VACCINE
Successfully completed in-life phase of GLP toxicology study in Sprague-Dawley rats. Final report not yet available, but no deaths or serious adverse events from myo-inositol inhalation reported to date.
Inhalation of dry powder measles vaccine by macaques in a pilot immunogenicity study was well tolerated with no deaths or adverse events reported. A robust measles-specific immune response was generated.
An inhalation toxicology study in additional Rhesus macaques is planned for next year.
Lam et al.* have completed a Phase I clinical trial of orally ingested myo-inositol powder with no SAEs reported following intake of up to 18 g per day for one month; Phase II clinical trials are beginning in Canada.
Normal concentration of myo-inositol in human plasma is about 4-5 mg/liter.
Intracellular concentration is 5-500x higher than in plasma.
Average dietary intake is 1 gram in the form of inositol hexaphosphate or myo-inositol in phospholipids.
Human milk contains about 450 mg/liter of myo-inositol.
*S. Lam, A. McWilliams, J. leRiche, C. MacAulay, L. Wattenberg, E. Szabo, "A Phase I Study of myo-Inositol for Lung Cancer Chemoprevention", Cancer Epidemiol Biomarkers Prev, 15 (8): 1526-1531, 2006.
Live-attenuated E-Z measles vaccine was reformulated as microparticles that rapidly dissolve in moist respiratory mucosa and become the functional equivalent of the wet-mist measles vaccine successfully administered to more than 3 million children.
Dried and micronized stabilized live virus measles vaccine without greater loss of viral activity than in the present commercial lyophilization process.
Pressure release valves or rupturable membranes facilitate dispersion and create stable aerosol clouds comparable to an FDA-approved active DPI.