Resources for Preclinical Resources for Preclinical Assessment of - - PowerPoint PPT Presentation

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

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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 t l & A t d d – Protocol & Assay standards – Inter-lab studies (ASTM, IANH, IRMM) – ASTM E56 & ISO TC 229

• Collaborative research

– NCTR: Primate studies – FDA: Dermal penetration of TiO2 – FDA: Nanosilver sterilization stability – NIEHS: Characterization studies

NCL is a resource to accelerate NCL is a resource to accelerate the clinical translation of the clinical translation of

• SAR studies

– Biocompatibility – Toxic mechanisms specific to nanoparticles Biological impact on size and surface characteristics

nanomedicines nanomedicines

– 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 Sh

In Vitro:

–Pharmacology –Blood contact properties –Shape –Composition –Molecular weight –Surface chemistry

In Vivo:

p p –Immune cell function –Cytotoxicity –Mechanistic toxicology St ilit –Surface chemistry –Identity –Purity –Stability –ADME –Safety –Efficacy –Sterility y –Solubility

http://ncl.cancer.gov/assay cascade.asp

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http://ncl.cancer.gov/assay_cascade.asp

Physicochemical Characterization Physicochemical Characterization

O O OH O O O N H O OH O OH O O O H H O HO O O

Small molecules

Physicochemical Parameters

Nanomaterial

• Elemental analysis
• Mass Spec
• NMR
• Composition
• Physical properties
• Chemical properties

Id tifi ti

• Microscopy (AFM,

TEM, SEM)

• Light scattering
• UV-Vis
• IR
• HPLC
• Identification
• Quality
• Purity
• Stability

g g (Static, Dynamic)

• SEC, FFF
• Electrophoresis
• GC
• Polarimetry

(CE, PAGE)

• Titration, Zeta
• Fluorimetry

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Same parameters – different/additional characterization methods

http://ncl.cancer.gov/working_assay-cascade.asp

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 – Hemolysis NCL Method ITA-1 y – Platelet Aggregation – Coagulation – Complement Activation Analysis of Hemolytic Properties of Nanoparticles

Nanotechnology Characterization Laboratory

p – CFU-GM – Leukocyte Proliferation – Macrophage/Neutrophil Function

Nanotechnology Characterization Laboratory National Cancer Institute at Frederick SAIC-Frederick Frederick, MD 21702 (301)-846-6939

– Cytotoxic Activity of NK Cells

• Toxicity

– Phase I/II Enzyme Induction/Suppression O S

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– Oxidative Stress – Cytotoxicity (necrosis) – Cytotoxicity (apoptosis)

http://ncl.cancer.gov/working_assay-cascade.asp

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

– Blood Chemistry

Plasma Profile

Dual Radiolabels

– Hematology – Histopathology – Gross Pathology

6 7 8 9 se 3H /mL 0.3 0.35 0.4 0.45 e 14C /mL 3H-Liposome 14C-Ceramide

Plasma Profile

Gross Pathology

• Efficacy

– Therapeutic

1 2 3 4 5 % Injected dos 0 05 0.1 0.15 0.2 0.25 % Injected dose

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– Imaging

http://ncl.cancer.gov/working_assay-cascade.asp

1 5 10 15 20 25 30 35 40 45 50 Time (hours) % 0.05 %

In Vivo Imaging

Optical Ultrasound MRI PET/CT Optical Ultrasound MRI PET/CT

X CRi Vi l S i Phili Si Xenogen Spectrum CRi Maestro GNIR-Flex Visual Sonics Vevo 770 Philips 3.0T

Archival System

Siemens Inveon µPET CT upgrade (6/08)

SPECT/CT High Resolution γ-Well Counter Archival System SPECT/CT High Resolution γ Well Counter

Si F ji P ki El Bi Siemens MIPORTAL Fuji FLA-5100 PerkinElmer 1480 Wizard Bioscan NanoSPECT/CT

Acknowledgements Acknowledgements

Scott E. McNeil, Ph.D. Christopher B. McLeland, B.S., M.B.A.

Nanotechnology Characterization Lab

, Marina A. Dobrovolskaia, Ph.D. Sarah Skoczen, M.S. Timothy M. Potter, B.S. Anil K. Patri, Ph.D. Stephan T. Stern, Ph.D., DABT p , , Barry W. Neun, B.S.

Supporting Staff

Marina A. Dobrovolskaia, Ph.D. Pavan Adiseshaiah, Ph.D. Sarah Skoczen, M.S. Matthew Hansen, M.S. Ruyin Shi M S Jeffrey D Clogston Ph D Sonny Man, M.S. Nick Panaro, Ph.D. Diana C. Haines, D.V.M., Diplomate ACVP Ulrich Baxa, Ph.D. Jennifer B. Hall, Ph.D. Ruyin Shi, M.S. Jamie Rodriguez, B.S. Jeffrey D. Clogston, Ph.D. Rachael M. Crist, Ph.D. David Parmiter, B.A. Sarah Anderson, B.S. Gloryvee Rivera, B.S. Melissa Orr B S , Contact Info: (301) 846-6939 Melissa Orr, B.S. Wendi Custer, B.A. ( ) ncl@mail.nih.gov http://ncl.cancer.gov

8 Funded by NCI Contracts N01-CO-12400 and HHSN261200800001E