Resources for Preclinical Resources for Preclinical Assessment of Nanomaterial Assessment of Nanomaterial Assessment of Nanomaterial Assessment of Nanomaterial Anil K. Patri Deputy Director, NCL patria@mail.nih.gov http://ncl.cancer.gov p g 1
NCL Objectives NCL Objectives NCL Objectives NCL Objectives • Preclinical characterization – Physico-chemical – In vitro In vitro – In vivo • Collaborative standards development – Nanoparticle RM standards with NIST – P Protocol & Assay standards t l & A t d d – Inter-lab studies (ASTM, IANH, IRMM) – ASTM E56 & ISO TC 229 Collaborative research • – NCTR: Primate studies – FDA: Dermal penetration of TiO 2 NCL is a resource to accelerate NCL is a resource to accelerate – FDA: Nanosilver sterilization stability the clinical translation of the clinical translation of – NIEHS: Characterization studies nanomedicines nanomedicines • SAR studies – Biocompatibility – Toxic mechanisms specific to nanoparticles – Biological impact on size and surface characteristics Biological impact on size and surface characteristics – Sterility & endotoxin • Education and knowledge sharing – Nanomedicine courses – Organize workshops and symposia – caNanolab web portal for data sharing
NCL Assay Cascade NCL Assay Cascade Physicochemical: –Size –Shape Sh In Vitro : –Composition –Pharmacology –Molecular weight In Vivo : –Blood contact properties p p –Surface chemistry –Surface chemistry –ADME –Immune cell function –Identity –Safety –Cytotoxicity –Purity –Mechanistic toxicology –Efficacy –Stability y –Sterility St ilit –Solubility http://ncl.cancer.gov/assay cascade.asp http://ncl.cancer.gov/assay_cascade.asp 3
Physicochemical Characterization Physicochemical Characterization O O O OH OH O O O O N H H OH O H HO O O O O O Physicochemical Small molecules Nanomaterial Parameters • Elemental analysis • Microscopy (AFM, • Composition • Mass Spec TEM, SEM) • Physical properties • Chemical properties • NMR • Light scattering g g • Identification Id tifi ti • UV-Vis (Static, Dynamic) • Quality • IR • SEC, FFF • Purity • HPLC • Stability • Electrophoresis • GC (CE, PAGE) • Polarimetry • Titration, Zeta • Fluorimetry Same parameters – different/additional characterization methods http://ncl.cancer.gov/working_assay-cascade.asp 4
In Vitro Cascade In Vitro Cascade • Sterility – Bacterial/Viral/Mycoplasma – Endotoxin – Endotoxin • Cell Uptake/Distribution – Cell Binding/Internalization – Targeting Targeting • Blood Contact Properties – Plasma Protein Binding NCL Method ITA-1 – Hemolysis y Analysis of Hemolytic – Platelet Aggregation Properties of Nanoparticles – Coagulation – Complement Activation p Nanotechnology Characterization Laboratory Nanotechnology Characterization Laboratory – CFU-GM National Cancer Institute at Frederick SAIC-Frederick – Leukocyte Proliferation Frederick, MD 21702 – Macrophage/Neutrophil Function (301)-846-6939 – Cytotoxic Activity of NK Cells • Toxicity – Phase I/II Enzyme Induction/Suppression – Oxidative Stress O S – Cytotoxicity (necrosis) – Cytotoxicity (apoptosis) http://ncl.cancer.gov/working_assay-cascade.asp 5
In Vivo Cascade In Vivo Cascade • Initial Disposition Study – Tissue Distribution – Clearance – Half-life • Immunotoxicity Immunotoxicity – 28-day screen – Immunogenicity (repeat dose tox study) study) • Dose-Range Finding Toxicity Dual Radiolabels – Blood Chemistry Plasma Profile Plasma Profile – Hematology 9 0.45 8 0.4 e 14 C /mL se 3 H /mL 3H-Liposome – Histopathology 7 0.35 14C-Ceramide 6 0.3 – Gross Pathology Gross Pathology % Injected dose % Injected dos 5 0.25 • Efficacy 4 0.2 3 0.15 – Therapeutic 2 0.1 % % 1 1 0.05 0 05 – Imaging 0 0 0 5 10 15 20 25 30 35 40 45 50 http://ncl.cancer.gov/working_assay-cascade.asp Time (hours) 6
In Vivo Imaging MRI MRI PET/CT PET/CT Optical Optical Ultrasound Ultrasound Phili Philips Siemens Si Xenogen X CRi CRi Vi Visual Sonics l S i 3.0T Inveon µPET Spectrum Maestro Vevo 770 CT upgrade (6/08) GNIR-Flex γ -Well Counter γ Well Counter SPECT/CT SPECT/CT High Resolution High Resolution Archival System Archival System Bi Bioscan PerkinElmer P ki El Fuji F ji Siemens Si NanoSPECT/CT 1480 Wizard FLA-5100 MIPORTAL
Acknowledgements Acknowledgements Nanotechnology Characterization Lab Scott E. McNeil, Ph.D. , Christopher B. McLeland, B.S., M.B.A. p , , Anil K. Patri, Ph.D. Timothy M. Potter, B.S. Stephan T. Stern, Ph.D., DABT Barry W. Neun, B.S. Supporting Staff Marina A. Dobrovolskaia, Ph.D. Marina A. Dobrovolskaia, Ph.D. Sarah Skoczen, M.S. Sarah Skoczen, M.S. Diana C. Haines, D.V.M., Nick Panaro, Ph.D. Sonny Man, M.S. Diplomate ACVP Pavan Adiseshaiah, Ph.D. Matthew Hansen, M.S. Ulrich Baxa, Ph.D. , Jeffrey D. Clogston, Ph.D. Jeffrey D Clogston Ph D Ruyin Shi M S Ruyin Shi, M.S. Sarah Anderson, B.S. Jennifer B. Hall, Ph.D. Jamie Rodriguez, B.S. Gloryvee Rivera, B.S. Rachael M. Crist, Ph.D. David Parmiter, B.A. Melissa Orr B S Melissa Orr, B.S. Wendi Custer, B.A. Contact Info: (301) 846-6939 ( ) ncl@mail.nih.gov http://ncl.cancer.gov Funded by NCI Contracts N01-CO-12400 and HHSN261200800001E 8
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