1. MEDICAL AEROSOLS TO IMPROVE GLOBAL HEALTH R.E. Sievers 1,2 , S. Winston, S.P. Cape 1,2 , D. Bennett 1 , J.L. Burger 2 , D.H. McAdams 2 , B.P. Quinn 1 , J.A. Searles 1 , D.M. Krank 1 , P. Pathak 1 , P. A. Bhagwat 1 , L.G. Rebits 1 , K.O. Kisich 3 , R. Dhere 4 , P. Kulkarni 4 , V. Vaidya 4 , R. Muley 4 , K. Powell 5 , C. Shermer 5 , L. Chan 5 , D. Griffin 6 , W-H. Lin 6 , P. Rota 7 , M. Papania 7 1 Aktiv-Dry LLC, 6060 Spine Road, Boulder, CO, USA 2 Center for Pharmaceutical Biotechnology, Department of Chemistry and Biochemistry, and CIRES, University of Colorado, Boulder, CO, USA 3 National Jewish Medical and Research Center, Denver, CO, USA 4 Serum Institute of India, Ltd., Pune, India 5 BD Technologies, Research Triangle Park, NC, USA 6 Johns Hopkins School of Public Health, Baltimore, MD, USA 7 Centers for Disease Control and Prevention, Atlanta, GA, USA

2. A lecture to honor Dr. Fred Fehsenfeld for all he has done for my group and for CIRES Sept.24, 2008

5. Henao, An Overview of Aerosol Immunization, Meeting of the WHO Steering Committee on New Delivery Systems, 2004

6. Henao-Restrepo and Papania, NIH Vaccines Conference, Rockville, Dec 2003

7. Dry Powder 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

13. Immunization in Difficult Circumstances ©Dr. Gordon Perkin Nepal 1999 Polio Outreach- Upper Nile region of Southern Sudan ©UNICEF ©UNICEF PATH

15. Measles Aerosol Vaccines in Humans 3 million humans immunized effectively with wet mist aerosol measles vaccine in Mexico (Valdespino, De Castro, et al.) Revaccination protection from mucosal pulmonary aerosol superior to SC injection in S. Africa (Dilraj, et al.) Nasal administration of large droplets gave poor seroconversion results in Chile (Simon, Levine, et al.) WHO Wet Mist Aerosol Group has been administering wet mists of SII Edmonston-Zagreb measles vaccine with 3 devices in Phase I clinical trials in India.

18. CAN-BD Nebulizer Drug Solution or Vaccine Suspension Near-critical or Supercritical CO 2 (80 to 100 atm) Flow Restrictor Tube ID = 75  m 10 cm Emulsion of product stream in liquid CO 2 CO 2 flashes upon rapid 100X decompression to atmospheric pressure Restrictor Tip

19. Tuberculosis Antibiotics Ciprofloxacin HCl Moxifloxacin HCl Rifampin Capreomycin Amikacin Drying T (C) 60 60 30 60 60 Solvent Water Water Ethyl Acetate Water Water Mean aero dia (µm) 1.2 1.1 0.78 1.3 1.2 95% cutoff (µm) 2.2 1.9 1.3 2.6 2.2 % < 5 um 89 56 86 77 84 Retained Activity 93% ± 20% 81% ± 43% 109% ± 34% 95% ± 7% 104% ± 7% AKTIV-DRY

20. Assembly: Raul and Mihesh

21. CAN-BD Generated Particles of Measles Vaccine Formulation Myo-inositol based placebo (M50)

23. Replication of Live Measles Virus Vaccine in Cotton Rat Lungs (K.O. Kisich)

25. CAN-BD Processing *Hydrophobic Vaccines Live attenuated measles virus Live attenuated influenza virus Hepatitis B surface antigen(HBsAg) Antibodies PRIMATZED® anti-CD4 Human IgG Anti-human lambda light chain Proteins Alpha-1-antitrypsin Trypsinogen Lactate dehydrogenase Lysozyme Insulin Alkaline phosphatase Nucleotides siRNA duplex Aptamers DNA single strand Formulation Constituents Sugars Buffers: Tricine, sodium potassium phosphate, sodium acetate Surfactants: Palmitic acid, stearic acid, Tween 20, Tween 80, Pluronic F68 Amino acids: Arginine, methionine, leucine, glycine, etc. Metal chelating agents: EDTA, DTPA Antibiotics Moxifloxacin Tobramycin Amoxycillin Doxycycline Cefazolin Ciprofloxacin Amikacin Capreomycin Rifampin Antivirals siRNA Relenza Other PEG PVP Hydrolyzed gelatin Sodium chloride DPPC (lung surfactant) Salbutamol (asthma drug) Albuterol sulfate Cromolyn sodium Phytosterols* (nutraceutical) Naproxen* Budesonide* Betamethasone* Amphotericin B* Cyclosporin* Myo-inositol and other sugars

32. “ GRAND CHALLENGE ” Bob Sievers Campan Marble Lourdes, France Gift installed in Norlin Historic Quadrangle at University of Colorado In gratitude for the support by the Foundation for the National Institutes of Health