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Tat-Conjugated PAMAM Dendrimers as Delivery Agents for Antisense and siRNA Oligonucleotides

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Purpose

PAMAM G5 dendrimer (P) was conjugated to Tat peptide (T), a cell penetrating peptide, in search of an efficient cellular delivery vehicle for antisense and siRNA oligonucleotides.

Methods

PAMAM G5 dendrimer was reacted with 4,4-difluoro-5,7-dimethyl-4-bora-3a,4a-diaza-s-indacene-3-propionic acid, sulfosuccinimidyl ester, sodium salt (BODIPY) for visualization to yield the conjugate BP. Bifunctional sulfosuccinimidyl 6-[α-methyl-α-(2-pyridyldithio)toluamido]hexanoate (sulfo-LC-SMPT) was then used to conjugate primary amino groups of BP to cysteine derivatized Tat peptide to give the designed conjugate, BPT. This conjugate was complexed with antisense and siRNA oligonucleotides designed to inhibit MDR1 gene expression. NIH 3T3 MDR cells were used for the evaluation of biological activity of the conjugate.

Results

Both antisense and siRNA readily formed complexes with the synthesized BPT, introduced into NIH 3T3 MDR cells, and primarily accumulated in intracellular vesicles. MDR1 gene expression was partially inhibited by the antisense–BPT complex and weakly inhibited by the siRNA–BPT complex when both were tested at nontoxic levels of dendrimer. Conjugation with Tat peptide did not improve the delivery efficiency of the dendrimer.

Conclusions

Dendrimer–oligonucleotide complexes were moderately effective for delivery of antisense and only poorly effective for delivery of siRNA. Conjugation of the dendrimer with the Tat cell penetrating peptide failed to further enhance the effectiveness of the dendrimer.

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Acknowledgments

The authors would like to thank Professor Kang Choon Lee of Sungkyunkwan University of Korea for helpful discussion on the use of sulfo-LC-SMPT. This work was supported by NIH grant PO1 GM59299 to R.L.J.

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Authors and Affiliations

  1. Department of Pharmacology, School of Medicine, University of North Carolina, Chapel Hill, North Carolina, 27599, USA

    Hyunmin Kang, Robert DeLong, Michael H. Fisher & Rudolph L. Juliano

  2. Department of Biology, Louisberg College, Louisberg, North Carolina, USA

    Robert DeLong

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  1. Hyunmin Kang
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  2. Robert DeLong
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  3. Michael H. Fisher
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  4. Rudolph L. Juliano
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Correspondence to Rudolph L. Juliano.

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Kang, H., DeLong, R., Fisher, M.H. et al. Tat-Conjugated PAMAM Dendrimers as Delivery Agents for Antisense and siRNA Oligonucleotides. Pharm Res 22, 2099–2106 (2005). https://doi.org/10.1007/s11095-005-8330-5

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  • DOI: https://doi.org/10.1007/s11095-005-8330-5

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