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Combination Nanopreparations of a Novel Proapoptotic Drug – NCL-240, TRAIL and siRNA

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Abstract

Purpose

To develop a multifunctional nanoparticle system carrying a combination of pro-apoptotic drug, NCL-240, TRAIL [tumor necrosis factor-α (TNF-α)-related apoptosis-inducing ligand] and anti-survivin siRNA and to test the combination preparation for anti-cancer effects in different cancer cells.

Methods

Polyethylene glycol-phosphoethanolamine (PEG-PE) – based polymeric micelles were prepared carrying NCL-240. These micelles were used in combination with TRAIL-conjugated micelles and anti-survivin siRNA-S-S-PE containing micelles. All the micelles were characterized for size, zeta potential, and drug encapsulation efficiency. Different cancer cells were used to study the cytotoxicity potential of the individual as well as the combination formulations. Other cell based assays included cellular association studies of transferrin-targeted NCL-240 micelles and study of cellular survivin protein downregulation by anti-survivin siRNA-S-S-PE containing micelles.

Results

NCL-240 micelles and the combination NCL-240/TRAIL micelles significantly increased cytotoxicity in the resistant strains of SKOV-3, MCF-7 and A549 as compared to free drugs or single drug formulations. The NCL-240/TRAIL micelles were also more effective in NCI/ADR-RES cancer cell spheroids. Anti-survivin siRNA micelles alone displayed a dose-dependent reduction in survivin protein levels in A2780 cells. Treatment with NCL-240/TRAIL after pre-incubation with anti-survivin siRNA inhibited cancer cell proliferation. Additionally, a single multifunctional system composed of NCL-240/TRAIL/siRNA PM also had significant cytotoxic effects in vitro in multiple cell lines.

Conclusion

These results demonstrate the efficacy of a combination of small-molecule PI3K inhibitors, TRAIL, and siRNA delivered by micellar preparations in multiple cancer cell lines.

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Abbreviations

c-FLIP:

Cellular FLICE (FADD-like IL-1β-converting enzyme)-inhibitory protein

EPR:

Enhanced permeability and retention

IAP:

Inhibitor of apoptosis

MDR:

Multi-drug resistant

mNCL-240:

NCL-240 – loaded polymeric micelles

mNCL-240/TRAIL:

NCL-240 – loaded, TRAIL-conjugated polymeric micelles

mSurvivin:

Anti-survivin siRNA-S-S-PE mixed micelles

mTf:

Transferrin – targeted polymeric micelles

PDK1:

3-phosphoinositide-dependent kinase-1

PE:

Phosphatidylethanolamine

PEG:

Polyethylene glycol

PH:

Pleckstrin-homology

PI3K:

Phosphoinositide 3-kinase

PIP2 :

Phosphatidylinositol-3,4-diphosphate

PIP3 :

Phosphatidylinositol-3,4,5-triphosphate

PKB/Akt:

Protein kinase B

PM:

Polymeric micelles

pNP:

p-nitrophenylcarbonyl

PTEN:

Phosphatase and tensin homologous protein

siRNA:

Small interfering RNA

siRNA-S-S-PE PM:

siRNA-S-S-PE – containing mixed micelles

Tf:

Transferrin

TRAIL:

Tumor necrosis factor-α (TNF-α)-related apoptosis-inducing ligand

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ACKNOWLEDGMENTS AND DISCLOSURES

NIH/NCI grant U54CA151881 to V.T.

Author information

Author notes

  1. Robert Riehle, Bhushan Pattni, Aditi Jhaveri, Abhijit Kulkarni, Ganesh Thakur & Vladimir Torchilin

    Present address: Center for Pharmaceutical Biotechnology and Nanomedicine, Department of Pharmaceutical Sciences, Northeastern University, 140 The Fenway, Room 236, 360 Huntington Avenue, Boston, Massachusetts, 02115, USA

Authors and Affiliations

  1. Department of Biochemistry, Tufts University School of Medicine, Boston, Massachusetts, USA

    Alexei Degterev

  2. Department of Biochemistry, Faculty of Science, King Abdulaziz University, Jeddah, 21589, Saudi Arabia

    Vladimir Torchilin

Authors
  1. Robert Riehle
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  2. Bhushan Pattni
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  6. Alexei Degterev
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  7. Vladimir Torchilin
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Corresponding author

Correspondence to Vladimir Torchilin.

Additional information

Robert Riehle and Bhushan Pattni contributed equally to this work.

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Riehle, R., Pattni, B., Jhaveri, A. et al. Combination Nanopreparations of a Novel Proapoptotic Drug – NCL-240, TRAIL and siRNA. Pharm Res 33, 1587–1601 (2016). https://doi.org/10.1007/s11095-016-1899-z

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  • DOI: https://doi.org/10.1007/s11095-016-1899-z

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