1. Kyung A. Kang Department of Chemical Engineering University of Louisville Britton Chance Symposium June 3-4, 2011 Growing-Up under Dr. Chance as Mentor and Thereafter: Near Infrared and Nanometal Particles for Molecular Imaging/Cancer Treatment

2. Working for Dr. Chance Working 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 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 Bio-heterogeneity (e.g., Tumor) Bio-heterogeneity (e.g., Tumor)

3. My Studies on NIR at PENN My Studies on NIR at PENN The Same Concept as Mass (or heat) Transfer in the System with Diffusion and Consumption (e.g., Oxygen Transport to Tissue). But it happens at a rate of 6 order of magnitude faster.The Same Concept as Mass (or heat) Transfer in the System with Diffusion and Consumption (e.g., Oxygen Transport to Tissue). But it happens at a rate of 6 order of magnitude faster. My Studies on NIR at PENN My Studies on NIR at PENN The Same Concept as Mass (or heat) Transfer in the System with Diffusion and Consumption (e.g., Oxygen Transport to Tissue). But it happens at a rate of 6 order of magnitude faster.The Same Concept as Mass (or heat) Transfer in the System with Diffusion and Consumption (e.g., Oxygen Transport to Tissue). But it 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 Method- Phased ArrayNew Bio-Heterogeneity Detection Method- Phased 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 Method- Phased ArrayNew Bio-Heterogeneity Detection Method- Phased Array

4. Systems Identification Systems Identification using Transfer Function, G(s)=Y(s)/X(s) Systems Identification Systems Identification using Transfer Function, G(s)=Y(s)/X(s)

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

6. Concept of NIR Phased Array

7. Personal Memories of Dr. Chance Dr. Chance’s 80 th BirthdayDr. Chance’s 80 th Birthday His Visit to University of Maryland for LectureHis Visit to University of Maryland for Lecture Dr. Chance’s 90 th BirthdayDr. Chance’s 90 th 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 Times)His Visit to Brown Cancer Center for Lecture (Two Times) His Visit to Louisville for the 36 th ISOTTHis Visit to Louisville for the 36 th ISOTT Visiting Dr. Chance Lecture Series in 2009Visiting Dr. Chance Lecture Series in 2009 Dr. Chance’s 80 th BirthdayDr. Chance’s 80 th Birthday His Visit to University of Maryland for LectureHis Visit to University of Maryland for Lecture Dr. Chance’s 90 th BirthdayDr. Chance’s 90 th 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 Times)His Visit to Brown Cancer Center for Lecture (Two Times) His Visit to Louisville for the 36 th ISOTTHis Visit to Louisville for the 36 th ISOTT Visiting Dr. Chance Lecture Series in 2009Visiting Dr. Chance Lecture Series in 2009

8. Dr. Chance at the 36 th ISOTT Conference in Louisville

9. Specific and Senstive Optical Contrast Agent using Gold Nanoparticles

10. 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 cm Depth Fluorescence Contrast By an ICG containing Absorber

11. Fluorescence and 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 Improve sensitivity

12. Distance control by polymer layer with known thickness Layer-by-layer polymer coating on GNP for distance control Polymer: PAH (+), PSS (-) Cypate conjugated polymer: PAHCy GNP: 10 nm PAH PSSPAHCy GNPGNP- (PAH/PSS)- PAHCy GNP- (PAH/PSS) GNP- PAH

13. Fluorophore 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 ICGCypate

14. Cypate fluorescence at various distance from GNP surface Control (Without GNP) Distance from GNP surface (nm) * **

15. Utilizing Both Quenching and Enhancement in One Entity? Distance from NGP surface Fluorescence

16. 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

17. Cancer cell Fluorescence quenched The complex being delivered to the cancer Ex Em Ex Em Ex Em

18. Cancer cell Some complexes targeting cancer cell Some are washed out by blood stream

19. Cancer cell Enzyme from cancer cell cleave short spacer

20. Cancer cell Short spacer is cut off -> long spacer determines the distance -> Fluorescence enhanced Em Ex Em Ex Em

22. 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)

23. Fluorescence with Long Spacers Long spacers and their estimated lengths by molecular simulation LinkersLength (nm) HS-(CH 2 ) 12 -PEG 4 -ONH 2 3.1 HS-(CH 2 ) 12 -PEG 6 -ONH 2 3.9 HS-(CH 2 ) 12 -PEG 8 -ONH 2 4.6 Cypate at 3.8 µM in water; GNP size, 10 nm

24. Nano-entity as Target Specific Thermal Guide

25. 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

26. AEM Field Generator & Probe (Dimension, 51x92x38 cm Power, 12.5 kW, Wt: 80 kg Frequency, 35-100, 135-400 kHz) Generator Probe of Common Configuration

27. Effect of Particle Size and Concentration on Heating Experimental Conditions: 1 ml sample; 300 KHz; ~3 KW; 5 minutes; solenoid coil. Size Concentration

28. Probe Configurations vs. Field Distributions Solenoid (most commonly used) Pancake Sandwich

29. 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

30. 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

31. 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

32. Primo Vascular System -32-      Organ surface BHC Skin BHD Skin BHC Intravascular Intravascular BHD BHD Brain BHD Brain BHD