Robert J Nicolosi - Director 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

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

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

Nanoemulsions of Tocopherol

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

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

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

Nanoemulsion delivery of anti- inflammatory agent Aspirin in mice

(JACS 2005)

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

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

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

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

Nanoemulsion delivery of Tamoxifen to breast cancer cell lines

Days of Culture The Effect of Tamoxifen (T) and a Nanoemulsion Preparation of Tamoxifen (NT) on Cell Proliferation C T NT

Dacarbazine Nano-emulsion Treatment in a Zenograft Melanoma Mouse Model

Control Dacarbazine Dacarbazine Topical IM Topical IM - 4% - 30% - 49% - 62% p < P < P < x 2x Tumor Reduction with Nano preparations of Dacarbazine Nano

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

Anti-Bacterial indication of Nanoemulsions

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

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

Summary Nanoemulsion technology produces particle sizes in the 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.

Acknowledgements Doctoral Students in the Biomedical Engineering & Biotechnology Program Jean-Bosco Tagne Srikanth Kakumanu Fongshu KuoBalaji Karthick Rohini VishwanathanMukta Bagul Microfluidics (MFIC) Corp, Newton MA 02469