Comparative Study of the Skin Penetration of Protein Transduction Domains and a Conjugated Peptide
- 513 Downloads
We examined the ability of a protein transduction domain (PTD), YARA, to penetrate in the skin and carry a conjugated peptide, P20. The results with YARA were compared to those of a well-known PTD (TAT) and a control, nontransducing peptide (YKAc). The combined action of PTDs and lipid penetration enhancers was also tested.
YARA, TAT, YKAc, P20, YARA-P20, and TAT-P20 were synthesized by Fmoc chemistry. Porcine ear skin mounted in a Franz diffusion cell was used to assess the topical and transdermal delivery of fluorescently tagged peptides in the presence or absence of lipid penetration enhancers (monoolein or oleic acid). The peptide concentrations in the skin (topical delivery) and receptor phase (transdermal delivery) were assessed by spectrofluorimetry. Fluorescence microscopy was used to visualize the peptides in different skin layers.
YARA and TAT, but not YKAc, penetrated abundantly in the skin and permeated modestly across this tissue. Monoolein and oleic acid did not enhance the topical and transdermal delivery of TAT or YARA but increased the topical delivery of YKAc. Importantly, YARA and TAT carried a conjugated peptide, P20, into the skin, but the transdermal delivery was very small. Fluorescence microscopy confirmed that free and conjugated PTDs reached viable layers of the skin.
YARA and TAT penetrate in the porcine ear skin in vitro and carry a conjugated model peptide, P20, with them. Thus, the use of PTDs can be a useful strategy to increase topical delivery of peptides for treatment of cutaneous diseases.
Key Words:P20 peptides skin topical delivery TAT YARA
Unable to display preview. Download preview PDF.
- 10.10. H. D. C. Smyth, G. Becket, and S. Mehta. Effect of permeation enhancer pretreatment on the iontophoresis of luteinizing hormone releasing hormone (LHRH) through human epidermal membrane (HEM). J. Pharm. Sci. 9:11296–11307 (2002).Google Scholar
- 18.18. C. R. Flynn, P. Komalavilas, D. Trssier, J. Thresher, E. E. Niederkofler, C. M. Dreiza, R. W. Nelson, A. Panitch, L. Joshi, and C. M. Brophy. Transduction of biologically active motifs of the small heat shock related protein HSP20 leads to relaxation of vascular smooth muscle. FASEB J. 17:1358–1360 (2003).PubMedGoogle Scholar
- 19.19. V. P. Torchilin and T. S. Levchenko. TAT-liposomes: a novel intracellular drug carrier. Curr. Prot. Pept. Sci 4:133–140 (2003).Google Scholar
- 20.20. S. Console, C. Marty, C. García-Escheverría, R. Schwendener, and K. Ballmer-Hefer. Antennapedia and HIV transactivator (TAT) “protein transaction domains” promote endocytosis of high molecular weight cargo upon binding to cell surface glycosaminoglycans. J. Biol. Chem. 278:35109–35114 (2003).PubMedGoogle Scholar
- 29.29. D. J. Tessier, P. Komalavilas, B. Liu, C. K. Kent, J. S. Thresher, C. M. Dreiza, A. Panitch, L. Joshi, E. Furnish, W. Stone, R. Fowl, and C. M. Brophy. Transduction of peptides analogs of the small heat shock-related protein HSP20 inhibits intimal hyperplasia. J. Vasc. Surg. 40:106–114 (2004).PubMedGoogle Scholar
- 33.33. K. Moser, K. Kriwet, A. Naik, Y. N. Kalia, and R. H. Guy. Passive skin penetration enhancement and its quantification in vitro. Eur. J. Pharm. Bipharm 52:103–112 (2001).Google Scholar
- 38.38. M. G. Carr, J. Corish, and O. I. Corrigan. Drug delivery from a liquid crystalline base across Visking and human stratum corneum. Int. J. Pharm. 157:35–42 (1997).Google Scholar
- 39.39. P. B. Robbins, S. F. Oliver, S. M. Sheu, P. Goodnough, P. Wender, and P. A. Khavari. Peptide delivery to tissues via reversibly linked protein transduction sequences. Biotechiques 33:190–194 (2002).Google Scholar
- 40.40. J. Park, J. Ryu, L. H. Jin, J. H. Bahn, J. A. Kim, C. S. Yoon, D. W. Kim, K. H. Han, W. S. Eum, H. Y. Kwon, T. C. Kang, M. H. Won, J. H. Kang, S. W. Cho, and S. Y. Choi. 9-Polylysine protein transduction domain: enhanced penetration efficiency of superoxide dismutase into mammalian cells and skin. Mol. Cells 13:202–208 (2002).PubMedGoogle Scholar
- 43.43. L. J. Weimann. J. Wu. Transdermal delivery of L-poly-lysine by sonomacroporation. Ultasound Med. Biol 28:1173–1180 (2002).Google Scholar
- 44.44. J. Y. Fang, W. R. Lee, S. C. Shen, H. Y. Wang, C. L. Fang, and C. H. Hu. Transdermal delivery of macromolecules by erbium: YAG laser. J. Control. Rel. 100:75–85 (2004).Google Scholar
- 46.46. T. F. Zioncheck, S. A. Chen, L. Richardson, M. Mora-Worms, C. Lucas, D. Lewis, J. D. Green, and J. Mordenti. Pharmacokinetics and tissue distribution of recombinant human transforming growth factor beta 1 after topical and intravenous administration in male rats. Pharm. Res. 11:213–220 (1994).PubMedGoogle Scholar
- 48.48. K. Lintner and O. Peschard. Biologically active peptides: from a laboratory bench curiosity to function skin care product. Int. J. Cosmet. Sci. 22:207–218 (2000).Google Scholar
- 50.50. H. Schaefer and T. E. Redelmeier. Skin Barrier. Principles of Percutaneous Absorption. Kaerger, Basel, 1996.Google Scholar