Robert J Nicolosi - Director

1. MicroFluidics Technology Fair Nanoemulsion Delivery of Pharmaceutical, Nutraceuticals and Cosmeceuticals In vitro and In vivo studies October 3, 2006 Center for Health and Disease Research Robert J Nicolosi - Director

2. Illustration of a Nano-Emulsion Nanoparticles (via AFM)) • The inner core consisting of any oil • Drug or nutrient molecules within the core • The emulsifier layer surrounding the nano-particle.

3. Characteristics of Nanoemulsions • Stable at temperatures up to 110° C and from pH < 2.0 – 8.0 • 6 month stability (at least) • Range of particle sizes (25-100 nm) • Converts fat soluble chemicals to stable water dispersions

4. Nanoemulsions of Tocopherol

5. Does Size make a difference in Efficacy of Delivering Tocopherol as a Micro (10-6) vs Nanoemulsion (10-9)

6. Vitamin E Enhancing Bioactivity: Microemulsion vs.NanoEmulsifion Reactive Oxygen Species (ROS) Nanoemulsions of Vitamin E are 100 times more active than Microemulsion Encapsulated Vitamin E in cell culture.

7. Transdermal Delivery of Tocopherol: Effect on Blood Levels in Hamsters (n=7) 3.7 ug/mL Tocopherol Tocopherol alone Nanoemulsion Of Tocopherol 2.09 ug/ml Tocopherol μg/mL 1 2 3 (Submitted 2006) Hours

8. Nanoemulsion delivery of anti- inflammatory agent Aspirin in mice

9. (JACS 2005)

10. “Anti-oxidant Synergy Formulation” suppresses malignancy: Nanoemulsions improve its effectiveness in culture Taxol • ASF forces differentiation of neuroblastoma (most common solid tumor in children)...Like neurons, if they begin to differentiate, they no longer multiply

11. ASF Studies (In vivo) • Nude Mice • since they lack a fully-functional immune system • Injection of Cancer Cells • by subcutaneous injection of Mouse or Human Cancer cells (300,000 cells /ml) • Injection of ASF solution • once tumors were detected • Measure the tumor diameter • quantified daily in accordance with the longest diameter

12. The neuroblastoma in the nude mice Tumor Control (no treatment) No Tumor ASF Nano-emulsion

13. Subcut Injection vs Topical Delivery of ASF containing- nanoemulsions on neuroblastoma–induced tumor growth rate. - 4% - 65% - 63% Subcut inj Subcut inj Transdermal ( In press 2006)

14. Nanoemulsion delivery of Tamoxifen to breast cancer cell lines

15. The Effect of Tamoxifen (T) and a Nanoemulsion Preparation of Tamoxifen (NT) on Cell Proliferation C T NT 1 3 5 7 10 12 Days of Culture

16. Dacarbazine Nano-emulsion Treatment in a Zenograft Melanoma Mouse Model

17. Tumor Reduction with Nano preparations of Dacarbazine p < 0.0007 P < 0.000003 P <0.0001 10x -4% 2x - 30% - 49% - 62% Nano Control Dacarbazine Dacarbazine Topical IM Topical IM

18. Tumor Growth in Mice after Dacarbazine Administration is Stopped for 12 Weeks + 43% + 30% Control Drug Topical - 18% - 23% - 92% Drug Nano-Drug IM Topical Nano-Drug IM

19. Anti-Bacterial indication of Nanoemulsions

20. Microemulsion versus Nanoemulsion of Soy Protein Nanoemulsified Homogenized Nanoemulsions are stable dispersions which do not agglomerate and which demonstrate antimicrobial activity

21. Gram + Staph Aureus Bacteria Staph Aureus Control Staph aureus streaked empty nanoemulsion

22. Summary • Nanoemulsion technology produces • particle sizes in the 25-100 nm range. • The technology converts fat soluble to • water soluble compounds. • The technology can deliver pharmaceuticals, • nutraceuticals and cosmeceuticals. • The particle size reduction is associated • with increased bioavailability and efficacy.

23. Acknowledgements Doctoral Students in the Biomedical Engineering & Biotechnology Program Jean-Bosco Tagne Srikanth Kakumanu Fongshu Kuo Balaji Karthick Rohini Vishwanathan Mukta Bagul Microfluidics (MFIC) Corp, Newton MA 02469