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Beta-casein Nanoparticles as an Oral Delivery System for Chemotherapeutic Drugs: Impact of Drug Structure and Properties on Co-assembly

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ABSTRACT

Purpose

To develop a novel oral drug delivery system comprising a hydrophobic chemotherapeutic drug entrapped within beta casein (β–CN), a major milk protein, which self-associates into micelles in aqueous solutions. The efficient gastric digestibility of β–CN suggests possible targeting to gastric cancers.

Methods

Antitumor drug entrapment was performed by stirring its dimethyl-sulfoxide solution into a phosphate-buffered saline containing β–CN. The association of drugs to β–CN was characterized by spectrophotometry and Trp143 fluorescence quenching; particle-size by dynamic light scattering, and colloidal stability by zeta potential.

Results

The optimal drug-to-β–CN molar loading-ratios for paclitaxel and vinblastine at 1 mg/ml β–CN were found to be 7.3 ± 1.2 and 5.3 ± 0.6 and the association constants were (6.3 ± 1.0)·103 M−1 and (2.0 ± 0.3)·104 M−1, respectively. Zeta potential analysis suggested that nanoencapsulation by β–CN stabilized all studied drugs in aqueous solution. The initial drug-β–CN association was apparently governed by hydrophobic interactions and at higher drug concentrations, also by electrostatic interactions. Up to the optimal drug:β–CN loading-ratio, ~80% of the particles were below 100 nm in diameter. At higher drug concentrations, particle diameter increased, and bi- or tri-modal particle distributions were observed.

Conclusions

Beta–CN forms colloidally-stable nanovehicles of hydrophobic anticancer drugs, and may be used for oral-delivery of chemotherapeutics.

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Abbreviations

β–CN:

beta casein

c log P:

calculated log P values

c pKa :

calculated pKa values

CMC:

critical micellization concentration

cryoTEM:

cryogenic transmission electron microscopy

DLS:

dynamic light scattering

DMSO:

dimethyl sulfoxide

F :

the fluorescence intensity at a given added ligand (drug) concentration

F 0 :

the fluorescence intensity at the beginning of the titration

F 1 :

the fluorescence intensity at the end of the titration

FRET:

fluorescence resonance energy transfer

GIT:

gastrointestinal tract

IV:

intravenous

K a :

the (average) apparent association constant of a single binding site

K d :

the (average) apparent dissociation constant of a single binding site

[LF]:

the molar concentration of the free ligand (drug)

log P:

log of the octanol-water partition coefficient

n:

number of binding sites per protein molecule

PBS:

phosphate buffer saline

[PF]:

free protein molar concentration

pI:

isoelectric pH

pKa :

negative log of the association constant (Ka)

[PL]:

the molar concentration of protein to which ligand is bound

Rg:

radius of gyration

Trp:

Tryptophan

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ACKNOWLEDGEMENTS

This work was supported by the generous donation of Yehuda and Civana Kahani, in memory of Aba Kahani, one of the pioneers of Israel’s Dairy Farming.

The Spectrofluorometer used herein was purchased with the support of the Russell Berry Nanotechnology Institute, Technion, Israel Institute of Technology.

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Authors and Affiliations

  1. Russell Berrie Nanotechnology Institute, Technion-Israel Institute of Technology, Haifa, 32000, Israel

    Alina Shapira & Yoav D. Livney

  2. The Fred Wyszkowski Cancer Research Laboratory Faculty of Biology, Technion-Israel Institute of Technology, Haifa, 32000, Israel

    Yehuda G. Assaraf

  3. The Laboratory of Biopolymers and Food Nanotechnology Faculty of Biotechnology & Food Engineering, Technion-Israel Institute of Technology, Haifa, 32000, Israel

    Dikla Epstein & Yoav D. Livney

Authors
  1. Alina Shapira
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  2. Yehuda G. Assaraf
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  4. Yoav D. Livney
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Corresponding author

Correspondence to Yoav D. Livney.

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Shapira, A., Assaraf, Y.G., Epstein, D. et al. Beta-casein Nanoparticles as an Oral Delivery System for Chemotherapeutic Drugs: Impact of Drug Structure and Properties on Co-assembly. Pharm Res 27, 2175–2186 (2010). https://doi.org/10.1007/s11095-010-0222-7

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