1. 1 Alternatives to Lyophilization AKTIV-DRY Jim Searles Ph.D. Aktiv-Dry LLC, Boulder, Colorado jsearles@aktiv-dry.com Visiongain 3rd Annual Lyophilization Conference Ensuring optimum formulations for pharmaceuticals and biologicals 25 February 2010, Boston

2. 2 Why consider alternatives to lyophilization? Capital and operating costs New routes of administration Dry powder inhalation Direct powder injection Value-added injection presentations Pre-filled reconstitution/injection devices Markets are shifting to single-dose primary contianers Powders are more versatile

3. 3 Product Attributes Particle size Reconstitution time Void volume Flowability Container Residual solvent (usually water) Product activity & stability Crystallinity Sterility Bulk intermediate or final drug product

4. 4 Aerosolization Aktiv-Dry PuffHaler®

6. Low void volume (avoids bubbles)Freeze drying not suitable

7. 6 Alternatives Discussed in This Presentation Conventional spray drying Bubble drying® Spray-freeze drying Stirred freeze drying Foam drying

8. 7 Process Attributes Mass yield Continuous or Batch? Product stresses Final product form Powder or cake Reconstitution Container Robustness Controllability Measurability Scale(s) available Closed system processing? Capital cost Operational cost

9. 8 Pharma Spray Drying Niro A/S

10. 9 Niro A/S Pharma Spray Drying

11. 10 Niro A/S Industrial Spray Drying

12. 11 Spray Drying

13. 12 Spray-Dried Products Pharmaceuticals Insulin (Exubera) Analgesics Antibiotics Enzymes Plasma/plasma substitutes Vaccines Vitamins Yeasts Speciality chemicals Catalysts Detergents Dyestuffs Fine organic/inorganic chemicals Foodstuffs and dairy products Baby food Cheese/ whey products Coconut milk Coffee/ coffee substitutes Coffee whitener Eggs Flavours Maltodextrine Mild Soup mixes Soy- based food Spices/ herb extracts Sugar-based food Tea Tomato Vegetable protein Ceramics Advanced ceramic formulations Carbides Ferrites Nitrides Oxides Silicates Steatites Titanates Polymers ABS e-PVC PMMA UF/MF resins Agro- chemicals Chelates Fungicides Herbicides Insecticides

14. 13 AKTIV-DRY F P T T Filters CO2 Product T Ctrl Drying Chamber P Drying Gas Flow Control T vF H2O vF CO2 Flow Control Pump N2 F Back-pulsed double filter Product Flow Control Pump Bubble Drying System

15. 14 Bubble Drying® Also known as Carbon dioxide assisted nebulization with a Bubble Dryer®(CAN-BD) Supercritical assisted atomization (SAA) Effervescent atomization Dry powder generation Respirable particle size (1-5 um for needle-free delivery) or larger Gentle low-temperature processing conditions Amenable to sterile, contained closed-system processing Dry powder delivery in the field promises improved consistency over liquid nebulization Gentler than jet milling Particle coating or co-particles Replacement for freeze drying & spray-freeze drying Wide Diversity of Product Types Vaccines, antibodies, proteins, siRNA’s, small-molecules

16. 15 AKTIV-DRY Flow Restrictor Tube ID = 75 m 10 cm Bubble Dryer Nebulizer Near-critical or Supercritical CO2 (80 to 100 atm) Drug Solution or Suspension Emulsion of product stream in liquid CO2 CO2 flashes upon rapid 100X decompression to atmospheric pressure Restrictor Tip

17. 16 AKTIV-DRY Bubble Dried Particles Same formulation lyophilized

18. 17 Bubble Drying Variations Non-aqueous solvents 2-pass operation Combination of 3 streams (cross) Coated particles Composite particles Slowly dissolving (PLA or PLGA) Readily soluble AKTIV-DRY

19. 18 Wide Range of Bubble Dried® Materials *hydrophobic

20. 19 AKTIV-DRY Bubble Dried Aqueous 10% NaCl

22. 21 Live Measles Vaccine 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 No greater loss of viral activity than in the present commercial lyophilization process Pass the WHO test for stability at 37 °C for one week First-ever dry powder inhaler for infants. Very low cost and complexity. Generates immune responses in Cotton rats and Rhesus macaques

23. 22 Spray Freeze Drying Process Spray freeze e.g. spray into liquid nitrogen Transfer into freeze dryer Freeze dry Not commercialized Effective for preservation of some material’s activity

24. 23 Alum-Adsorbed Vaccines Maa et al. 2003 J Pharm Sci. 92(2):319-332 Hepatitis-B surface antigen (Alum-HBsAg) and Diphtheria and tetanus toxoids (Alum-DT) Freeze drying, spray drying, air drying, or spray freeze drying Only spray freeze drying yielded fully active vaccine Rapid freezing key Bubble Drying® has also yielded fully active, very stable flowable powders of alum-adsorbed HBsAg (Sievers et al. J Supercrit Fluids 42(3):385-39)

25. 24 Other SFD examples Split-virion and sub-unit influenza vaccines Maa et al. 2004 J Pharm Sci 93(7):1912 - 1923 Anthrax vaccine Jiang et al. J Pharm Sci 95(1):80-96 Inulin-stabilized influenza subunit vaccine Amorij et al. 2007 Vaccine 25:8707-8717 Recombinant Human Interferon- Webb et al. 2002 J Pharm Sci 91:1474-1487

26. 25 Hosokawa “Stirred freeze drying comes down to injecting the material to be dried into the vessel, where it gets into contact with the freezing medium (usually carbon dioxide) and immediately solidifies in to granular material.” “Subsequently a vacuum is created in the machine, the product is kept in motion and the mixture is dried. Because sublimation occurs while the product is in motion, more vapour comes to the surface and the drying process evolves rapidly.”

27. 26 Foam Drying Investigated for proteins and vaccines Key References Victor Bronshtein 1998 patent Patents by Vu Truong-Le, MedImmune Vaccines (‘06, ‘08) Two 2007 papers by Abdul-Fattah et al. Pisal et al. 2006 paper Reconstitution time?

29. Place vials on the freeze dryer shelf at 15 C, and allow to equilibrate for 10 min

30. Reduce the chamber pressure to 50 mTorr

31. Initiates foaming

32. Hold for 24 h for primary drying

34. Process loss similar to lyophilization

36. 30 Monoclonal Antibody Abdul-Fattah et al. 2007 J Pharma Sci 96(8), 1983-2008 IgG1 Freeze drying, spray drying, and foam drying “Preparations manufactured by foam drying were the most stable, regardless of the stabilizer level”

37. 31 Stability at 25 ºC from square-root-of-time kinetics Note: Bubble Dried® measles vaccine at 37 ºC: 0.8 log/wk0.5 Abdul-Fattah et al. 2007 Pharmaceutical Research 24(4) 715-727

38. Another Direct Comparison 32 Live attenuated measles vaccine Foam Lyo SD SD+AD Vaccine. 2010 Feb 3;28(5):1275-84.

39. 33 Acknowledgements Aktiv-Dry Brian Quinn Scott Winston Pankaj Pathak Pradnya Bhagwat Lia Rebits Dave Krank University of Colorado Prof. Bob Sievers Steve Cape Joseph Villa John Carpenter David McAdams Jessica Burger