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Targeted Delivery of Complexes of Biotin–PEG–Polyethylenimine and NF-κB Decoys to Brain-derived Endothelial Cells in Vitro

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Abstract

Purpose

To evaluate the effect of re-directing the uptake mechanism of polyplexes containing oligodeoxynucleotide (ODN) decoys to nuclear factor kappa B (NF-κB) from absorptive-mediated to receptor-mediated endocytosis.

Materials and Methods

Complexes of ODNs and a co-polymer of biotin–polyethylenglycol and polyethylenimine (BPP) were targeted to brain-derived endothelial cells with a conjugate of antibody 8D3 and streptavidin (8D3SA). Size and stability of ODN/BPP complexes was measured by dynamic light scattering. Cellular uptake was studied by confocal microscopy. Cell viability and pharmacological effects were investigated on murine bEnd5 cells stimulated with tumor necrosis factor.

Results

ODN/BPP complexes showed sizes of 116 ± 2.3 nm, which increased by 40 nm when coupled to 8D3SA, and were stable in physiological fluids. Targeted complexes were internalized intact into endosomal compartments. Treatment conditions, which yielded significant inhibitory effects on mRNA expression of VCAM-1, ICAM-1, IκBα and iNOS by bEnd5 cells, did not affect viability. At 0.5 μM, decoy ODN significantly inhibited monocyte adhesion to bEnd5 monolayers when delivered as 8D3SA-targeted complex, while higher concentrations of untargeted complex were ineffective.

Conclusions

The complex of NF-κB decoys and BPP, which can be targeted to transferrin receptors, is a promising drug candidate for neuroinflammatory diseases affecting the blood–brain barrier.

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Abbreviations

BBB:

blood–brain barrier

BCECF-AM:

2′,7′-bis-(carboxyethyl)-5-(6)-carboxyfluorescein-acetoxymethyl ester

BPP:

biotin–polyethylenglycol–polyethylenimine

ICAM-1:

intercellular adhesion molecule 1

LMW-PEI:

low molecular weight polyethylenimine

NF-κB:

nuclear factor kappaB

ODN:

oligodeoxynucleotide

PEG:

poly (ethylene glycol)

PEI:

polyethylenimine

SA:

streptavidin

S-SMPB:

sulfosuccinimidyl 4-[p-maleimidophenyl]butyrate

VCAM-1:

vascular cell adhesion molecule 1

8D3SA:

8D3-streptavidin

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Acknowledgments

The authors thank Dr. Holger Petersen (Basel) for synthesis and analysis of the biotin-PEG-PEI copolymer and Young Tag Ko for preparation of the rhodamine-biotin-PEG-PEI conjugate. We appreciate the helpful discussions with Dr. Thomas Kissel (Marburg). This work was supported by grant 1R01NS045043 to UB.

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Authors and Affiliations

  1. Department of Pharmaceutical Sciences, Texas Tech University Health Sciences Center School of Pharmacy, 1300 Coulter Drive, Amarillo, Texas, 79106, USA

    Raktima Bhattacharya, Berit Osburg, Dagmar Fischer & Ulrich Bickel

  2. Department of Pharmaceutics and Biopharmacy, Philipps-University of Marburg, Marburg, Germany

    Dagmar Fischer

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  1. Raktima Bhattacharya
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  2. Berit Osburg
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  3. Dagmar Fischer
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  4. Ulrich Bickel
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Correspondence to Ulrich Bickel.

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Bhattacharya, R., Osburg, B., Fischer, D. et al. Targeted Delivery of Complexes of Biotin–PEG–Polyethylenimine and NF-κB Decoys to Brain-derived Endothelial Cells in Vitro . Pharm Res 25, 605–615 (2008). https://doi.org/10.1007/s11095-007-9389-y

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