Molecular Imaging and Therapy with
Perfluorocarbon Nanoparticulates:
Environmental Impact.
Washington University Medical School
Saint Louis, MO
GENERALIZED TARGETING PARADIGM
Payloads
Imaging agents (Gd3+)
Drugs
Radionuclides
Anti-fibrin
Anti-tissue factor
Anti−αvβ3-integrin
19F
Lipid coat
Gd3+
(MAb, aptamers, small peptides, peptidomimetics, polysaccharides, etc)
Perfluorocarbon Biomedical Applications
Liquid Ventilation
Vitreous
Replacement
Fibrin
Imaging
Scanning EM
After
Before
Targeted
Contrast
Fibrin
RBC
clot labeled with
fibrin targeted
nanoparticles
0.5 µ
0.5 µ
0.5 µ
Clot
Clot
Ultrasound
contrast
enhancement of fibrin
clot in vitro using
conventional clinical
scanner (7.5 MHz)
Suture
1 mm
1 mm
Suture
Control Thrombus Targeted Thrombus
Targeted Nanoparticles Support
Enormous Paramagnetic Payloads
ανβ3-Targeted
Non-Targeted
94,400
92,400
r1 (s*mM)-1 [Gd]
19.1
21.1
r2 (s*mM)-1 [Gd]
22.9
24.6
mM)-1 [Particle]
r1 (s*mM)
1,800,000
1,950,000
r2 (s*mM)-1 [Particle]
2,160,000
2,270,000
Gd3+ Ions / Particle
Detection of Angiogenesis in the Vx-2 Model
Masses Seen By MRI Were Not Always
Vx-2 Tumors
T2-Weighted MRI
After
avB3-Nanoparticles
5 mm
Histology Revealed Rejected Tumor
Infiltrated with Inflammatory Cells
No Molecular Imaging of
Angiogenesis for Tumors in
Rejection
Histology Revealed Rejected Tumor
Infiltrated with Inflammatory Cells
Melanoma Angiogenesis: Detection With αvβ3
Integrin-Targeted Paramagnetic Nanoparticles
Mouse Imaging @ 1.5T
Time 0 min
syringe
2 mm
tumor
Time 120 min
Angiogenesis around
2 mm melanoma tumor
Time course
Time course of MRI Contrast Enhancement
Percent Enhancement
180
160
140
1
Targeted
1
J
Control
H
Competition
1
1
120
1
100
J
80
60
J
1
H
J
H
40
20
01HJ
0
J
J
H
H
H
20
40
60
80
Time (min) Post Injection
100
120
Site-Targeted Drug Delivery
1. Load with drug
Target Cell
3. Drug effect
D
D
2. Particles bind to cell surface
Targeting Element
0.5 µ
D
Novel Mechanisms Of Drug Delivery:
“Contact Facilitated Drug Delivery*”
Rhodamine-labeled NP
Drug Loaded Outer
Lipid Monolayer
Exchange of
Lipids and Drug
Components
Epitopes
Target Cell Membrane
*Lanza GM, Wickline SA et al: Targeted antiproliferative
GFP-linked endocytotic markers
drug delivery to vascular smooth muscle cells with magnetic
resonance imaging nanoparticle contrast agent: implications rab5 and rab7 in transfected
for rational therapy of restenosis. Circulation 2002; 106:2842C32 melanoma cells
2847
Antiproliferative Effects of Free versus Tissue
Factor - Targeted Paclitaxel Nanoparticles on
Vascular Smooth Muscle Cells
CELL COUNT X 10,000
30
25
20
15
*
10
*
5
0
Control
Taxol 0.2 mol %
Taxol 2.0 mol %
NANOPARTICLE FORMULATION
NON -TARGETED NANOPARTICLES
TARGETED NANOPARTICLES
Rejection of Mouse Melanoma 7d following ανβ3-Targeted,
DXR, Paramagnetic Nanoparticles
Control
3/3
ανβ3−DXR-NP
(4/4)
Proton and Fluorine Imaging
Clot
Proton Imaging
Fluorine Imaging
Dosimetry of Targeted Nanoparticle Levels
15:5 Crown-Ether
Bound volume ( µ L)
40
y = 0.2665x - 2.1857
R2 = 0.9462
30
20
10
0
-10
0
20
40
60
80
Emulsion percentage
100
PFOB
Bound Emulsion Volume
(µL)
6
y = 0.0493x - 0.641
4
R 2 = 0.9178
2
0
-2
0
25
50
75
Estimated % of Emulsion Volume
100
Typical Pharmaceutical Emulsification Process
Alliance Pharmaceutical
Oxygent Production
President Bush: reduce the greenhouse gas
intensity of the U.S. economy by 18% over the next
ten years.
Perfluorocarbons have 10,000 times the global
warming impact of carbon dioxide
Major PFC applications:
• electronic insulators
• aluminum smelting
• refrigerants
• subsurface barrier testing
EPA suggested four options for its
formal definition of "leak rate." :
• "annualizing" method.
• "rolling-average" method
• the higher calculated leak rate.
• consistent method
Non-VOC with no significant participation
in ozone formation in ambient air.
WU Personnel
Medical
Gregory Lanza, MD, PhD
Samuel Wickline, M.D.
MR & US Science
Patrick Winter, Ph.D.
Michael Hughes, Ph.D
Jon Marsh, Ph.D
Frank Hockett, M.S.E.E.
Michal Lijowski PhD
Grace Hu, M.S.
Anagana Pan Ph.D
Technical
Mary Watkins, R.T. (MR)
Todd Williams, B.S., R.T. (MR)
Rosa Lin, M.S.
Brian Hennery, B.S.
Ralph Fuhrhop
Huiying Zhang, M.D.
Stacy Allen, B.S.
Liz Lacy, B.S.
Peggy Brown, RDCS
Michael Scott, BS
Anne Schmeider, M.S.
Kathy Crowder, MS
Key Collaborators
Philips Medical Systems
The Dow Chemical Co.
David Rollo MD
Jaime Simon PhD
Horace Hines PhD
Garry Kiefer PhD
Ling Shao PhD
Phillip Athey PhD
Carmen van Vilsteren PhD
Gyongyi Gulyas PhD
Christopher Hall PhD
Keith Frank PhD
Shelton Caruthers PhD
Bristol-Myers Squibb
Medical Imaging
D. Scott Edwards
Stephen Haber
Thomas Harris
Consultants
Sam Achilefu PhD
Wynn Volkert PhD
Richard Holmes PhD
Patrick Gaffney PhD