Measuring Exposure Levels of Drug Products Containing Nanomaterials

Katherine Tyner, Ph.D.

CDER/OPQ

US Food and Drug Administration

July 8, 2015

1

Disclaimer

This talk reflects the views of the author and should not be construed to represent FDA’s views or policies The mention of commercial products, their sources, or their use in connection with material reported herein is not to be construed as either an actual or implied endorsement of such products by the Department of Health and Human Services.

2

Agenda

• Considerations for intentional exposure studies

• In vitro examples

• In vivo examples

3

Agenda

• Considerations for intentional exposure studies

• In vitro examples

• In vivo examples

4

An Intentional Discussion • In the majority of instances, the administration of drug products

containing nanomaterials means an intentional exposure – Unintentional exposure is possible and is part of a robust risk assessment

• Know the exposure has occurred

– Material – Dose – Pharmacokinetics/Toxicity (PK/TOX) – ADME (adsorption, distribution, metabolism, elimination)

• Detection is still key

– In vitro – In vivo models – Clinical

5

6

Platform

Example Name NDA

Approval Indication

Liposome DOXIL® (Doxorubicin) 19951 Cancer

Inorganic nanoparticle FERRLECIT® (Sodium ferric gluconate complex) 19992 Anemia

Protein nanoparticle ABRAXANE® (Paclitaxel) 2005 Cancer

Polymer nanoparticle MACUGEN® (Pegaptanib sodium) 2004 Macular degeneration.

Emulsion RESTASIS® (Cyclosporine) 2002 To increase tear production

Lipid complex AMPHOTEC® (Amphotericin B) 1996 Invasive aspergillosis

Nanotube SOMATULINE DEPOT® (Lanreotide acetate) 2007 Acromegaly

Nanocrystal TRICOR® (Fenofibrate) 20043 Hypercholesterolemia

Micelle TAXOTERE®(Docetaxel) 1996 Cancer

1 First ANDA approval in 2013 2 First ANDA approval in 2011 3 First ANDA approval in 2011

Nanomaterials in Drug Products: CDER Examples

Tyner KT et al. WIRES Nanomedicine and Nanotechnology 2015.

Analytical Methods for Detecting Nanomaterials in Biological Systems • Non-carbon nanomaterials have methodology for

quantification and characterization • Detection of elemental signal & visual confirmation • Example: ICPMS, TEM/EDS

7

?

Untreated animal

Analytical Methods for Detecting Nanomaterials in Biological Systems

8

PK/ADME/Toxicity

• Pharmacokinetics (PK) is the study of the kinetics of absorption, distribution, metabolism and excretion of drugs and their pharmacologic, therapeutic or toxic response in animals and man

• ADME – Adsorption

• How it gets in the body – Distribution

• Where it goes in the body – Metabolism

• How the body breaks it down – Elimination

• How the body gets rid of it

9

PK/ADME Methods & Nanomaterials

• Analytical methods – Most popular techniques for small molecules are radiolabeling and bioanalytical

techniques – Radiolabeling or fluorescently tagging nanomaterials may alter biodistribution

• Common bioanalytical techniques are not always valid for nanomaterials

– Example: Nanomaterials interacting with filtration step or chromatography columns

• Variety of matrices – Blood, urine, feces, on and off-target tissues

• Questions/considerations

– What is being measured/labeled (drug vs carrier)? – Does the nanomaterial remain intact?

• How are the constituents being identified – Is the bioanalytical method appropriate?

• Controls • Testing conditions

Before

After

10

Detection and Quantitation of Nanomaterials in Biological Matrices

• HPLC-MS – Dendrimers, Polymers, Drugs, Metabolites, Fullerenes

• Capillary Electrophoresis

– Fullerenes

• Radiolabeling – Liposomes, Dendrimers, Polymers

• ICP-MS & Neutron Activation Analysis

– Metallic Nanoparticles (e.g., Gold) – Metal Oxides

• Electron Microscopy

– EDS, EELS (Detection and confirmation of composition)

• Optical Microscopy – Raman, Hyperspectral imaging (Detection)

11

Serum

Serum + Dendrimer

Agenda

• Considerations for intentional exposure studies

• In vitro examples

• In vivo examples

12

In Vitro Examples: Is the Method Appropriate? Controls

Hyperspectral Imaging Positive id

Naïve cells

13

14

- Mass balance & control of exposure

- Cell culture conditions influence assay results

- Artificial constraints and/or parameters may influence results

Bancos S et al. J Nanobiotechnology 2015, 12

Static-based culture conditions Inserts-based culture conditions

Suspension-based culture conditions

In Vitro Examples: Is the Method Appropriate? Design Considerations

Agenda

• Considerations for intentional exposure studies

• In vitro examples

• In vivo examples

15

In Vivo Examples

16

0.0%0.5%1.0%1.5%2.0%2.5%3.0%3.5%

0 500 1000 1500

min

% ID

/mL

TNF-Au NP PK in rats, [TNF] by ELISA; [Au] by ICP-MS

Stern S.T. et. al. J. Control Release, 2010, 146, 164-174.

In Vivo Example: What is Being Measured? Complementary Analysis

17

0123456789

0 5 10 15 20 25 30 35 40 45 50Time (hours)

% In

ject

ed d

ose

3 H /m

L

00.050.10.150.20.250.30.350.40.45

% In

ject

ed d

ose

14C

/mL3H-Liposome

14C-Ceramide

Vd (mL*kg-1)

Mean ± SD

Ceramide 1020 ± 478

Liposome 63 ± 19

In Vivo Example: Does the Nanomaterial Remain Intact? Dual Labeling

Zolnik, B.S. et. al. Drug Metab Dispos, 2008, 36, 1709-1715 18

In Vivo Example: Is the Method Appropriate? Capillary Electrophoresis

Chan, K.C.; Patri, A.K.; Veenstra, T.D.; McNeil, S.E.; Issaq, H.J.; Electrophoresis; 2007, 28, 1518-1524 19

0100200300400500600700800

urineday 1

fecesday 1

urineday 4

fecesday 4

mg/

kg S

i

SiO2 NPControl

- Detection - Limits of detection - Limits of quantification - Nanomaterial interference - Appropriate controls - Well-designed studies

20

In Vivo Example: Is the Method Appropriate? Testing Considerations

Conclusions

• PK/ADME determination for drug products containing nanomaterials is an evolving area

• Techniques & methods exist to detect nanomaterials within biological tissues

– Multiple endpoints/methods

• Multiple issues and/or considerations may confound method development and analysis

– Appropriate controls – Well designed studies

• Regulatory science projects continue to address these issues

– Collaboration opportunities!

21

Acknowledgements

• Dr. Anil Patri, Director, NCTR-ORA Nanotechnology Core Facility

• The Nanotechnology Characterization Laboratory

• The authors would like to acknowledge the FDA White Oak Nanotechnology Core Facility for instrument use, scientific and technical assistance

22