Irfan_Khan_42X40_Poster_ICC 20151010

INTRODUCTION:Material-guided drug delivery technology utilizing magnetic nanoparticles has emerged as the next generation tool for cancer treatment. Targeted drug delivery can significantly reduce undesirable side effects and raise the therapeutic index of a drug. Nanovectors coupled with doxorubicin or doxorubicin prodrugs, functionalized via bio-conjugation chemistry, are able to access cancerous cells, as assessed by confocal microscopy and FACS analysis. Mutually orthogonal chemistries were utilized to attach doxorubicin, targeting groups, as well the fluorescent dyes to the nanoparticles. Click-to-release strategy has been employed for doxorubicin release upon cellular nanoparticle internalization.

“Bio-orthogonal conjugation of doxorubicin to magnetic nanoparticles as targeted delivery system”

Irfan Khan and Maksim Royzen The RNA Institute and Department of Chemistry, University at Albany-SUNY, NY 12222, USA

Methods:1. Nanoparticle synthesis 2. Prodrug of Doxorubicin synthesis 3. Cytotoxicity assay, standard MTT assay was used for

determination of IC50 in MDA-MB-231 cells treated with Dox or Dox-R-TCO

4. Fluorescence confocal microscopy was performed to visualize the internalization of conjugated magnetic nanoparticles in MDA-MB-231 cells

5. Caspase 3 activity assay, conventional Caspase 3 assay was used for measurement of apoptosis in MDA-MB-231 cells treated with bio-orthogonal moieties conjugated magnetic nanoparticles

6. FACS analysis, was used to measure cellular uptake of nanoparticle

MNP

Prodrug activation

Prodrug

Results:1. The IC50 of Doxorubicin and Prodrug Dox-TCO measured as 0.098 and 0.48 µM respectively. 2. We measured caspase-3 activity in cell lysates after 96 h of incubation with tetrazine conjugated magnetic nanoparticles and Doxorubicin or Dox-TCO attached with releasable triggers. The concentration of Doxorubicin and Dox-R-TCO used were (0.75 uM) and (2 uM) respectively. The strongest statistically pronounced effect was observed in cells treated with Conjugated-MNP’s Cy5.5 treated cells as compared to No MNP’s control or MNP’s Cy5.5 control. 3. Fluorescence confocal microscopy revealed the internalization of conjugated magnetic nanoparticles in MDA-MB-231 cells4. FACS results revealed particular sorting of specific flourochrome labeled cells treated with bio-orthogonal moieties conjugated magnetic nanoparticles.

Doxorubicin: IC50=0.098 µM

Dox-TCO: IC50=0.48 µMConclusion:Our results overall suggest that, the magnetic nanodrugs can be used for “Image-guided” or targeted drug delivery in breast cancer cells.

References:http://www.nanomedicine.dtu.dk/Research/Gold.aspx http://www.healthcare.siemens.com/magnetic-resonance-imaging/0-35-to-1-5t-mri-scanner/magnetom-essenza/use

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) Caspase-3 activity assay