1. 1 NCL and caNanoLab http://ncl.cancer.gov 5 th Annual Meeting June 23-25, 2008 Washington DC

2. 2 NCL provides infrastructure support to the Alliance and to nanotech researchers - to overcome obstacles and translate ‘nano’ into the clinical realm. NCL Objectives –Characterize nanoparticles using standardized methods –Conduct structure activity relationships studies –Facilitate regulatory review of nanotech constructs –Engage in educational and knowledge sharing efforts The NCL is a national resource available to investigators from academia, industry and government NCI Cancer Nanotechnology Plan ncl@ncifcrf.gov http://ncl.cancer.gov

3. 3 NCL Concept of Operations NCL is a formal collaboration between NCI, FDA and NIST

4. 4 In Vitro: –Pharmacology –Blood contact properties –Immune cell function –Cytotoxicity –Mechanistic toxicology –Sterility Physicochemical: –Size –Shape –Composition –Molecular weight –Surface chemistry –Identity –Purity –Stability –Solubility In Vivo: –ADME –Safety –Efficacy http://ncl.cancer.gov/assay_cascade.asp NCL Assay Cascade

5. 5 Elemental analysis Mass Spec NMR UV-Vis IR HPLC GC Polarimetry Small molecules Physicochemical Parameters Composition Physical properties Chemical properties Identification Quality Purity Stability Same parameters – different/additional characterization methods Microscopy (AFM, TEM, SEM) Light scattering (Static, Dynamic) SEC, FFF Electrophoresis (CE, PAGE) Zeta sizer Fluorimetry Nanomaterial Physical Characterization http://ncl.cancer.gov/working_assay-cascade.asp

6. 6 Sterility –Bacterial/Viral/Mycoplasma –Endotoxin Cell Uptake/Distribution –Cell Binding/Internalization –Targeting Blood Contact Properties –Plasma Protein Binding –Hemolysis –Platelet Aggregation –Coagulation –Complement Activation –CFU-GM –Leukocyte Proliferation –Macrophage/Neutrophil Function –Cytotoxic Activity of NK Cells Toxicity –Phase I/II Enzyme Induction/Suppression –Oxidative Stress –Cytotoxicity (necrosis) –Cytotoxicity (apoptosis) NCL Method ITA-1 Analysis of Hemolytic Properties of Nanoparticles Nanotechnology Characterization Laboratory National Cancer Institute at Frederick SAIC-Frederick Frederick, MD 21702 (301)-846-6939 http://ncl.cancer.gov/working_assay-cascade.asp In Vitro Cascade

7. 7 Initial Disposition Study –Tissue Distribution –Clearance –Half-life Immunotoxicity –28-day screen –Immunogenicity (repeat dose tox study) Dose-Range Finding Toxicity –Blood Chemistry –Hematology –Histopathology –Gross Pathology Efficacy –Therapeutic –Imaging http://ncl.cancer.gov/working_assay-cascade.asp In Vivo Cascade Plasma Profile Dual Radiolabels

8. 8 Dendritic Nanotechnologies (DNT) Avidimer Therapeutics M. Brechbiel (NCI) E. Simanek (Texas A&M) M. Kester (Penn State) E. Chang (Georgetown) R. Blumenthal (NCI) J. Nagy (NanoValent) J. Connor (Penn State) W. Zamboni (U. Pittsburgh) A. Miller (Imperial College London) NanoSpectra (Rice) S. Weiss (UCLA) Evident Technologies J. Barchi (NCI) US FDA TiO 2 Liposomes Gold nanoshells Fullerenes Quantum Dots Dendrimers M. Amiji (Northeastern) Nanoscan Nanoemulsions CytImmune Sciences TedPella Inc. A. Wei (Purdue) K. Katti (U. Missouri) Colloidal gold Polymers J. Ljubimova (Mt. Sinai) M. Amiji (Northeastern) N. Tarasova (NCI) D. Ferguson (U. Wisc) V. Torchilin (Northeastern) Carigent Therapeutics Tego Bioscience Alnis/Aduro Iron Oxide Portfolio of Nanoparticles

9. 9 Develop Protocols Protocols are developed and validated at the NCL. Intent is to leverage existing methods and/or Standards when possible. Submit Draft Standard Draft of Standard is submitted to ASTM E-56.20 Subcommittee for comment and revision STANDARD E5620-06 Ballot of Standard is submitted to ASTM members for consensus. Consensus Ballot Solicit Comments on Protocol Protocols are submitted to partners and collaborators for comment and revision. NCI FDA NIST Voluntary Consensus Standard (VCS) is established. Industry, Academia, Government VCS informs regulatory agencies and promotes commercialization of nanotechnology for medical applications NCL Method ITA-1 Analysis of Hemolytic Properties of Nanoparticles Nanotechnology Characterization laboratory National Cancer Institute at Frederick SAIC-Frederick Frederick, MD 21702 (301)-846-6939 Voluntary Consensus Standards

10. 10 First voluntary consensus standards for biocompatibility- testing of nanomaterials intended for medical applications E2524 (hemolysis), E2525 (CFU-GM inhibition), E2526 (kidney and liver cytotox) ASTM Standards RMs NCI supported the production of NIST’s colloidal gold RM. –Au selected for calibration and biocompatibility 10nm, 30nm, and 60 nm diameters Available as of January ’08 Standard Reference Materials Standard Methods Interlaboratory Comparison ASTM ILS 166 Standards Development

11. 11 Data Dissemination http://ncl.cancer.gov Conferences and Working Groups Newsletters Manuscripts and Book Chapters Formal Reports

12. 12 Data Dissemination CaNanoLab provides storage and retrieval of nanoparticle characterizations for data sharing in support of NCL’s educational and knowledge sharing efforts.

13. 13 Informatics & Science –Data management (storage and retrieval) –Data sharing –Comparison of multiple results (e.g. size and toxicity) –Provide the necessary input into Structure Activity Relationship (SAR) modeling and simulation tools

14. 14 caNanoLab: Physical Characterization

15. 15 caNanoLab: In Vitro Characterization

16. 16 NCL Scott McNeil, Ph.D. Anil Patri, Ph.D.Marty Fritts, Ph.D. Marina Dobrovolskaia, Ph.D. Stephan Stern, Ph.D. Jeffrey D. Clogston, Ph.D.Jiwen Zheng, Ph.D. Parag Aggarwal, Ph.D.Hari Devalapally, Ph.D. Jennifer Hall, Ph.D.Sarah Skoczen, M.S. Chris McLeland, B.S.Tim Potter, B.S. Barry Neun, B.S.Jamie Rodriguez, B.S. David Parmiter, B.A.Nathan Whye, B.S. Funded by NCI Contract N01-CO-12400 Acknowledgements Contact Info: ncl@mail.nih.gov 301-846-6939 http://ncl.cancer.gov