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Purpose.
Topical delivery of oligonucleotides, though attractive for the treatment of skin disorders, is limited by the low permeability of the stratum corneum. Herein, we assessed the potential of low-frequency ultrasound (20 kHz, 2.4 W/cm2) in delivering therapeutically significant quantities of anti-sense oligonucleotides into skin.
Methods.
Dermal penetration of oligonucleotides penetration was quantified in vitro using radiolabeled oligonucleotides.
Results.
Estimated concentrations of oligonucleotides (ODNs) in the superficial layers of the skin ranged from ~0.5% to 5% of the donor concentration after a 10-min application of ultrasound and ODN. Microscopic evaluations using fluorescently labeled oligonucleotides and sulforhodamine B revealed heterogeneous penetration into the skin. Heterogenous penetration led to the formation of localized transport pathways (LTPs), which occupied about 5% of the total exposed skin area. Immuno-histochemical studies using an oligonucleotide that reacts specifically with an antibody also confirmed penetration of ODNs into LTPs. Histologic studies revealed that no gross structural changes were induced in the skin due to ultrasound application.
Conclusions.
These results show successful delivery of anti-sense oligonucleotides using low-frequency ultrasound.
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References
1. G. Jarad, J. S. Simske, J. R. Sedor, and J. R. Schelling. Nucleic acid-based techniques for post-transcriptional regulation of molecular agents. Curr. Opin. Nephrol. Hpertens. 12:415–421 (2003).
2. C. Wraight and P. White. Antisense oligonucleotides in cutaneous therapy. Pharmacol. Ther. 90:89–104 (2001).
3. U. Mrowietz. The enigma of cyclosporin A treatment for psoriasis: systemic efficacy versus topical non-responsiveness. A review. Acta Derm. Venereol. 72:321–326 (1992).
4. F. P. Schmook, A. Stutz, and J. Reinhardt. Penetration of Sand-immune (cyclosporin A) in rat skin in vitro. Effects of penetration enhancers and solvents. Skin Pharmacol. 6:116–124 (1993).
5. J. Bouwstra, A. M. de Vries, A. G. Gooris, S. W. Bras, J. Brussee, and M. Ponec. Thermodynamic and Structural Aspects of the Skin Barrier. J. Control. Rel. 15:209–220 (1991).
6. R. M. Brand and P. L. Iversen. Iontophoretic delivery of atelomeric oligonucleotide. Pharm. Res. 13:851–854 (1996).
7. K. Li, A. H. Ghanem, C. L. Teng, G. E. Hardee, and W. I. Higuchi. Iontopohoretic transport of oligonucleotides across human epidermal membrane: a study of the Nernst-Planck model. J. Pharm. Sci. 90:915–931 (2001).
8. R. M. Brand, T. L. Hannah, J. Norris, and P. L. Iversen. Transdermal delivery of antisense oligonucleotides can induce changes in gene expression in vivo. Antisense Nucleic Acid Drug Dev. 11:1–6 (2001).
9. V. Regnier, N. De Morre, A. Jadoul, and V. Preat. Mechanisms of a phosphorothioate oligonucleotide delivery by skin electroporation. Int. J. Pharm. 184:147–156 (1999).
10. T. E. Zewert, U. F. Pliquett, R. Langer, and J. C. Weaver. Transdermal transport of DNA antisense oligonucleotides by electroporation. Biochem. Biophys. Res. Commun. 212:286–292 (1995).
11. R. C. Mehta, K. K. Strecker, S. R. Cooper, M. V. Templin, Y. J. Tsai, T. P. Condon, C. F. Bennett, and G. E. Hardee. Intercellular adhesion moleculae-1 supression in skin by topical delivery of anti-sense oligonucleotides. J. Invest. Dermatol. 115:805–812 (2000).
12. W. Lin, M. Cromier, A. Samiee, A. Griffin, B. Johnson, C.-L. Teng, G. E. Hardee, and P. E. Daddona. Transdermal delivery of antisense oligonucleotides with microprojection patch (Macroflux) technology. Pharm. Res. 18:1789–1793 (2001).
13. S. Mitragotri, D. Edwards, D. Blankschtein, and R. Langer. A mechanistic study of ultrasonically enhanced transdermal drug delivery. J. Pharm. Sci. 84:697–706 (1995).
14. S. Mitragotri and L. Le. Transdermal delivery of heparin and low-molecular weight heparin using low-frequency ultrasound. Pharm. Res. 18:1151–1156 (2000).
15. B. P. Monia, J. F. Johnston, D. J. Ecker, M. A. Zounes, W. F. Lima, and S. M. Freier. Selective inhibition of mutant Ha-ras mRNA expression by antisense oligonucleotides. J. Biol. Chem. 267:19954–19962 (1992).
16. B. P. Monia, E. A. Lesnik, C. Gonzalez, W. F. Lima, D. McGee, C. J. Guinosso, A. M. Kawasaki, P. D. Cook, and S. M. Freier. Evaluation of 2′-modified oligonucleotides containing 2′-deoxy gaps as antisense inhibitors of gene expression. J. Biol. Chem. 268:14514–14522 (1993).
17. A. Tezel, A. Sens, J. Tuchscherer, and S. Mitragotri. Frequency dependence of sonophoresis. Pharm. Res. 18:1694–1700 (2001).
18. N. P. Katz, D. E. Shapiro, T. E. Hermann, J. Kost, and L. Custer. Rapid onset of cutaneous anesthesia with EMLA cream after pretreatment with a new ultrasound emitting device. Anesth. Analg. 98:371–376 (2004).
19. M. Butler, K. Stecker, and C. F. Bennet. Cellular distribution of phosphorothioate oligodeoxynucleotides in normal rodent tissue. Lab. Invest. 77:379–388 (1997).
20. S. Mitragotri, D. Blankschtein, and R. Langer. Ultrasound-mediated transdermal protein delivery. Science 269:850–853 (1995).
21. A. Tezel, A. Sens, and S. Mitragotri. Description of transdermal transport of hydrophilic solutes during low-frequency sonophoresis based on a modified porous pathway model. J. Pharm. Sci. 92:381–393 (2003).
22. W. Lin, M. Cormier, A. Samiee, A. Griffin, B. Johnson, C. L. Teng, G. E. Hardee, and P. E. Daddona. Transdermal delivery of antisense oligonucleotides with microprojection patch (Macroflux) technology. Pharm. Res. 18:1789–1793 (2001).
23. S. K. Li, A. H. Ghanem, C. L. Teng, G. E. Hardee, and W. I. Higuchi. Iontophoretic transport of oligonucleotides across human epidermal membrane: a study of the Nernst-Planck model. J. Pharm. Sci. 90:915–931 (2001).
24. V. Regnier, N. De Morre, A. Jadoul, and V. Preat. Mechanisms of a phosphorothioate oligonucleotide delivery by skin electroporation. Int. J. Pharm. 184:147–156 (1999).
25. A. Tezel, A. Sens, J. Tuchscherer, and S. Mitragotri. Frequency dependence of sonophoresis. Pharm. Res. 18:1694–1700 (2001).
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Tezel, A., Dokka, S., Kelly, S. et al. Topical Delivery of Anti-sense Oligonucleotides Using Low-Frequency Sonophoresis. Pharm Res 21, 2219–2225 (2004). https://doi.org/10.1007/s11095-004-7674-6
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DOI: https://doi.org/10.1007/s11095-004-7674-6