Kyung A. Kang

Department of Chemical Engineering

University of Louisville

Britton Chance SymposiumJune 3-4, 2011

Growing-Up under Dr. Chance as Mentor and Thereafter:

Near Infrared and Nanometal Particles for

Molecular Imaging/Cancer Treatment

Growing-Up under Dr. Chance as Mentor and Thereafter:

Near Infrared and Nanometal Particles for

Molecular Imaging/Cancer Treatment

Working for Dr. ChanceWorking for Dr. Chance Working for Dr. ChanceWorking for Dr. Chance

• Time: August 1991-January 1994Time: August 1991-January 1994 (Between Drs. E. Sevick and H. Liu)(Between Drs. E. Sevick and H. Liu)

• Near Infrared Spectroscopy:Near Infrared Spectroscopy: - Oxygen Saturation- Oxygen Saturation

- Detection & Characterization of- Detection & Characterization of Bio-heterogeneity (e.g., Tumor) Bio-heterogeneity (e.g., Tumor)

• Time: August 1991-January 1994Time: August 1991-January 1994 (Between Drs. E. Sevick and H. Liu)(Between Drs. E. Sevick and H. Liu)

• Near Infrared Spectroscopy:Near Infrared Spectroscopy: - Oxygen Saturation- Oxygen Saturation

- Detection & Characterization of- Detection & Characterization of Bio-heterogeneity (e.g., Tumor) Bio-heterogeneity (e.g., Tumor)

My Studies on NIR at PENNMy Studies on NIR at PENN•The Same Concept as Mass (or heat) Transfer in the System with The Same Concept as Mass (or heat) Transfer in the System with Diffusion and Consumption (e.g., Oxygen Transport to Tissue). But it Diffusion and Consumption (e.g., Oxygen Transport to Tissue). But it happens at a rate of 6 order of magnitude faster.happens at a rate of 6 order of magnitude faster.

My Studies on NIR at PENNMy Studies on NIR at PENN•The Same Concept as Mass (or heat) Transfer in the System with The Same Concept as Mass (or heat) Transfer in the System with Diffusion and Consumption (e.g., Oxygen Transport to Tissue). But it Diffusion and Consumption (e.g., Oxygen Transport to Tissue). But it happens at a rate of 6 order of magnitude faster.happens at a rate of 6 order of magnitude faster.

• Breast Cancer Detection/CharacterizationBreast Cancer Detection/Characterization (With Radiology Department)(With Radiology Department)

• New Analysis Approach for TRS dataNew Analysis Approach for TRS data

• New Bio-Heterogeneity Detection MethodNew Bio-Heterogeneity Detection Method - - Phased ArrayPhased Array

• Breast Cancer Detection/CharacterizationBreast Cancer Detection/Characterization (With Radiology Department)(With Radiology Department)

• New Analysis Approach for TRS dataNew Analysis Approach for TRS data

• New Bio-Heterogeneity Detection MethodNew Bio-Heterogeneity Detection Method - - Phased ArrayPhased Array

Systems Systems IdentificationIdentificationusing Transfer Function, G(s)=Y(s)/X(s)using Transfer Function, G(s)=Y(s)/X(s)

Systems Systems IdentificationIdentificationusing Transfer Function, G(s)=Y(s)/X(s)using Transfer Function, G(s)=Y(s)/X(s)

Frequency Response Analysis of TRS data (time domain->frequency domain)

Concept of NIR Phased ArrayConcept of NIR Phased ArrayConcept of NIR Phased ArrayConcept of NIR Phased Array

Personal Memories of Dr. ChancePersonal Memories of Dr. ChancePersonal Memories of Dr. ChancePersonal Memories of Dr. Chance

•Dr. Chance’s 80Dr. Chance’s 80thth Birthday Birthday

•His Visit to University of Maryland for LectureHis Visit to University of Maryland for Lecture

•Dr. Chance’s 90Dr. Chance’s 90thth Birthday Birthday

•His Visit to University of Louisville for LectureHis Visit to University of Louisville for Lecture•His Visit to Brown Cancer Center for Lecture (Two His Visit to Brown Cancer Center for Lecture (Two Times)Times)

•His Visit to Louisville for the 36His Visit to Louisville for the 36 thth ISOTT ISOTT

•Visiting Dr. Chance Lecture Series in 2009Visiting Dr. Chance Lecture Series in 2009

•Dr. Chance’s 80Dr. Chance’s 80thth Birthday Birthday

•His Visit to University of Maryland for LectureHis Visit to University of Maryland for Lecture

•Dr. Chance’s 90Dr. Chance’s 90thth Birthday Birthday

•His Visit to University of Louisville for LectureHis Visit to University of Louisville for Lecture•His Visit to Brown Cancer Center for Lecture (Two His Visit to Brown Cancer Center for Lecture (Two Times)Times)

•His Visit to Louisville for the 36His Visit to Louisville for the 36 thth ISOTT ISOTT

•Visiting Dr. Chance Lecture Series in 2009Visiting Dr. Chance Lecture Series in 2009

Dr. Chance at the 36Dr. Chance at the 36thth ISOTT ISOTT Conference in LouisvilleConference in Louisville

Dr. Chance at the 36Dr. Chance at the 36thth ISOTT ISOTT Conference in LouisvilleConference in Louisville

Specific and Senstive Optical Contrast Agent using Gold

Nanoparticles

ICG, FDA approved Fluorophore: Ex/Em, 780/830 nm; 2 cm S-D separation, Modulation frequency of 0.1 GHz. Heterogeneity - vitamin E capsule

Absorber 1.0 cm 1.5 cm 2.0 cm 2.5 cmDepth

Fluorescence ContrastBy an ICG containing Absorber

Fluorescence Fluorescence and Gold Nanoparticleand Gold NanoparticleFluorescence Fluorescence and Gold Nanoparticleand Gold Nanoparticle

Distance

Fluorescence

When a fluorophore is placed near a particle, fluorescence can be altered depending upon the field strength (i.e., the distance from the particle).

Quenching Enhancement

negative sensing/ molecular beacon

negative sensing/ molecular beacon

Improve sensitivity

Improve sensitivity

Distance control by polymer layer Distance control by polymer layer with known thicknesswith known thickness

Distance control by polymer layer Distance control by polymer layer with known thicknesswith known thickness

Layer-by-layer polymer coating on GNP for distance control

Polymer: PAH (+), PSS (-)

Cypate conjugated polymer: PAHCy

GNP: 10 nm

PAH PSS PAHCy

GNP GNP-(PAH/PSS)-

PAHCy

GNP-(PAH/PSS)

GNP-PAH

Fluorophore of our choice, CypateFluorophore of our choice, CypateFluorophore of our choice, CypateFluorophore of our choice, Cypate

FDA approved, Indocyanine Green (ICG) based, NIR fluorophore. Ex/Em, 780/830 nm (developed by Dr. Achileful at the Washington University)

Near infrared (NIR): Deep penetration in tissue Better differentiated from autofluorescence

FDA approved, Indocyanine Green (ICG) based, NIR fluorophore. Ex/Em, 780/830 nm (developed by Dr. Achileful at the Washington University)

Near infrared (NIR): Deep penetration in tissue Better differentiated from autofluorescence

ICG Cypate

Cypate fluorescence at various Cypate fluorescence at various distance from GNP surfacedistance from GNP surface

Cypate fluorescence at various Cypate fluorescence at various distance from GNP surfacedistance from GNP surface

Control (Without GNP)

0

5

10

15

20

0 1 2 3 4 5 6 7 8 9 10

Distance between from GNP surface (nm)

Rel

ativ

e F

luor

esce

nce

(Fol

d)

Distance from GNP surface (nm)

**

****

Utilizing Both Utilizing Both Quenching and Enhancement Quenching and Enhancement

in One Entity?in One Entity?

Utilizing Both Utilizing Both Quenching and Enhancement Quenching and Enhancement

in One Entity?in One Entity?

Distance from NGP surface

Fluorescence

Optical Contrast Agent, NanoPPET (Nanometal Particle Plasmon Energy Transfer)

(with Drs. Nantz and Achilefu)

Optical Contrast Agent, NanoPPET (Nanometal Particle Plasmon Energy Transfer)

(with Drs. Nantz and Achilefu)

Molecule for cancer target

NGP

Fluorophore

Structure to be cleaved by Cancer secreting enzyme

Excitation Light Fluorescence

Short Linker for fluorescence quenching

Long Linker for fluorescence enhancement

Cancer cell

Fluorescence quenchedThe complex being delivered to the cancer

ExEm

Em

Ex

Ex

Em

Ex

Em

Cancer cell

Some complexes targeting cancer cell

Some are washed out by blood stream

Cancer cell

Enzyme from cancer cell cleave short spacer

Cancer cell

Short spacer is cut off -> long spacer determines the distance -> Fluorescence enhanced

Em

Ex

Em

Ex

Ex

Em

NGPNGP

Cypate

Plasmon field

PEG

PEG with G-G-R peptide sequence

4-Hydroxytamoxifen

Molecular Beacon like Molecular Beacon like NanoparticleNanoparticle SystemSystem(Our Model System)(Our Model System)

Cancer cell Urokinase type

Plaminogen Activator

Urokinase Sensitive Short Spacer: Quenching and Restoring

Samples in 0.01M PBS buffer; Cypate Conc., 12.8 µM; GNP size, 10 nm; GNP concentration, 3.0 nM

Fluorescence Quenching by GNP

Restoration of Fluorescence by Cleaving Spacer by Urokinase (UPA)

Our Short Spacer is GGGRGG, 2.5 nm (theoretical estimation)

Fluorescence with Long Spacers

Long spacers and their estimated

lengths by molecular simulation

Linkers Length (nm)

HS-(CH2)12-PEG4-ONH2 3.1

HS-(CH2)12-PEG6-ONH2 3.9

HS-(CH2)12-PEG8-ONH2 4.6

Cypate at 3.8 µM in water; GNP size, 10 nm

Nano-entity as Target Specific

Thermal Guide

AEM field for Hyperthermia - Magnetic Nanoparticle as Thermal Guide

Minimal normal tissue heating Non-invasive heating Relatively Low cost Magnetic nanoparticles can be heated by

AEM field -> Cancer specific heating

AEM Field Generator & Probe

(Dimension, 51x92x38 cmPower, 12.5 kW, Wt: 80 kgFrequency, 35-100, 135-400 kHz)

Generator

Probe of Common Configuration

Effect of Particle Size and Concentration on Heating

Experimental Conditions: 1 ml sample; 300 KHz; ~3 KW; 5 minutes; solenoid coil.

Size of the nanoparticles, nm

0 10 20 30 40 50 60

C

-2

0

2

4

6

8

10

12

14

16

0.01 wt%0.03 wt%0.05 wt%0.08 wt%0.1 wt%

Concentration, wt.%

0.00 0.02 0.04 0.06 0.08 0.10 0.12

C

-2

0

2

4

6

8

10

12

14

16

10 nm20 nm30 nm50 nm

Size Concentration

Probe Configurations vs. Field Distributions

Solenoid (most commonly used)

Pancake

Sandwich

Tumor Specific Dual-functional Nano-entity

Gold coated magnetic nanoparticle for both optical detection and hyperthermia (and for x-ray& MRI contrast)

Biocompatible polymer layer

Tumor specific biomolecules -> for tumor target

Spacer

Fluorophore -> for cancer detection

Injection of PF127/MNP/anticancer drug mixture into tumor

Instant gel formation at body temperature

Sustained drug release

Hyperthermia and sustained drug release

heat

Nanoparticle/Temperature Sensitive Polymer for Hyperthermia and sustained drug release

To me, Dr. Chance was:To me, Dr. Chance was:

• Ｃｕｒｉｏｕｓ• Positive• Research/Work• Communicate with Anyone• Minimally Materialistic• Help Others behind the Scene

• Ｃｕｒｉｏｕｓ• Positive• Research/Work• Communicate with Anyone• Minimally Materialistic• Help Others behind the Scene

Primo Vascular SystemPrimo Vascular System

-32-