PhotoImmuno-NanoTherapy (PINT)

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• PhotoImmuno-NanoTherapy (PINT)
• Mark Kester
• G. Thomas Passananti Professor of Pharmacology
• Director, Penn State Center for NanoMedicine and
  Materials

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PhotoImmuno-NanoTherapy (PINT) Mark
Kester Keystone Nano, Inc State College,
PA Chief Medical Officer
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PhotoImmuno-NanoTherapy (PINT) Mark
Kester Keystone Nano, Inc State College,
PA Chief Medical Officer
Harnessing the Power of Light for Theranostics
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NanoJacketsCalcium Phosphate NanoParticles
(CPNPs)
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Nanojackets are Molecular Smart Bombs
Encapsulated components are released as a
function of pH
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TEM of ICG-Doped CPNPs of 16nm mean diameter
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• Limitations of free ICG
• Fluorescence instability in physiological
  environments
• Dimerization leads to fluorescent quenching
• Protein binding causes absorption shifts
• Rapid elimination from the body
• Plasma t1/2 3-4 minutes
• Taken up exclusively by hepatic parenchymal cells
• Subsequently secreted entirely into the bile
• Benefits of NanoJacket Encapsulation
• Monomer caging prevents aggregate formation
• Solvent protection improves photostability
• Surface passivation affords long-term in vivo
  circulation
• Tumor localization via the EPR-Effect

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Colloidal Stability of CPNPs in PBS
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Absorption and Fluorescence Spectra of Free ICG
and ICG-Doped CPNPs in Aqueous Solution
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Fluorescence Lifetime of Free ICG and ICG-CPNPs
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Comparative Spectral Effects of Various Solvents
on the Emission Response of Free ICG and ICG-CPNPs
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EPR effect
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In Vivo Administration of PEGylated ICG-CPNPs

• Accumulation within tumors via enhanced
  permeation retention effect
• Internalization into tumor cells via endosome
  pathway

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Comparative Fluorescence Signal Intensity as
Function of Depth in Porcine Muscle Tissue
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Active Targeting
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Theory of Photodynamic Therapy

• Requirements photosensitizer, light, and oxygen
• Photosensitizer is excited at appropriate
  wavelength from ground singlet state to excited
  singlet state
• Excited photosensitizer undergoes intersystem
  crossing to excited triplet state
• Energy transfer to molecular oxygen (ground
  triplet state) allows photosensitizer to relax to
  ground singlet state
• Oxygen is now in excited singlet state
• Singlet oxygen is highly reactive

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Current Limitations

• Low quantum yield of photosensitizers
• Short lifetimes of photosensitizers
• Lack of specific targeting
• Inability to penetrate sufficient light to
  targets
• Poor understanding of mechanisms linking singlet
  oxygen generation to cancer cell death

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Photodynamic Therapy Utilizing ICG-Loaded
Calcium Phosphate Nanoparticles Prevents Breast
Cancer Growth In Vivo (Single IV Injection and
Single NIR Treatment)
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PINT Extends Survival of Leukemic Mice (3 IV
Injections and Splenic NIR Treatment)
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In Vitro Photodynamic Therapy With ICG-CPNPs
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Myeloid Derived Suppressor Cells Decrease
5-Days Post-NIR Treatment
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PINT Reduces MDSCs in 410.4 Tumor-Bearing BALB/cJ
Mice
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PINT Increases Natural Killer Cells in MDA-MB-231
Tumor-Bearing Nude Mice
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PINT Increases Dendritic and Natural Killer Cells
in 410.4 Tumor-Bearing BALB/cJ Mice
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Adoptive Transfer
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Injection of Culture-(PINT) Treated Cells in
Tumor-Established Nude Mice
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Conclusion

• Nanotechnology has the potential to deliver the
  promise of light-based pharmaceutics

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Acknowledgements

• Brian Barth
• Tom Stover
• James Kaiser
• Todd Fox
• Onar Unal
• Yasser Haekal
• Tony Brown
• Sean ONeil
• Kristie Houck
• Murali Nagarajan
• Sriram Shamnugavelandyu
• Lindsey Rylund
• Tom Loughran
• Xin Liu

• James Adair
• Sarah Rouse
• Erhan Antinouli
• Tom Morgan
• Peter Eklund
• Peter Butler
• MK is Founder and CMO for Keystone Nano, Inc
• NanoJackets have been licensed by Penn State
  Research Foundation to Keystone Nano, Inc., State
  College, PA