Abstract
Purpose. This study investigates the structure/activity relationship of a series of N-acyl-peptides (lipopeptides) for the transfection of mammalian cells.
Methods. Lipopeptides comprising 1 to 3 basic amino-acids and a single fatty acid chain were synthesized. Transfecting complexes between lipopeptide, plasmid DNA and dioleoyl phosphatidylethanolamine were prepared and applied on cells in culture. Transfection efficiency was evaluated by measuring β-galactosidase activity 48 h post-transfection. Lipopeptide-DNA binding was also investigated by physical means and molecular modelling.
Results. Besides the length of the fatty acid chain, the nature of the basic amino-acid and the C-terminal group were crucial parameters for high transfection efficiency. The N-acyl-(diaminobutyric acid)n derivatives were the most potent transfecting agents among those tested and induced a β-galactosidase activity 2 to 20 times higher than the N-acyl-lysine, -ornithine or -diaminopropionic acid derivatives. Furthermore, a hydrazide C-terminal modification greatly enhanced transfection efficiency for all compounds tested. The reason why α, γ-diaminobutyric acid hydrazide-based lipopeptides were the most potent in transfection is not fully understood but could be related to their high DNA binding.
Conclusions. Poly- or oligo-diaminobutyric acid containing or not a hydrazide C-terminus could advantageously be used in peptide-based gene delivery systems.
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REFERENCES
J. P. Behr. Bioconjugate Chem. 5:382–389 (1994).
X. Gao and L. Huang. Gene Ther. 2:710–722 (1995).
P. L. Felgner, T. R. Gadek, M. Holm, R. Roman, H. W. Chan, M. Wenz, J. P. Northrop, G. M. Ringold, and M. Danielsen. Proc. Natl. Acad. Sci. USA 84:7413–7417 (1987).
J. Y. Legendre and F. C. Szoka. Pharm. Res. 9:1235–1242 (1992).
J. Zabner, A. J. Fasbender, T. Moninger, K. A. Poellinger, and M. J. Welsh. J. Biol. Chem. 270:18997–19007 (1995).
J. S. Remy, C. Sirlin, P. Vierling, and J. P. Behr. Bioconjugate Chem. 5:647–654 (1994).
Vogler, P. Lanz, P. Quitt, R. O. Studer, W. Lergier, E. Böhni, and B. Fust. Helv. Chim. Acta 47:526–544 (1964).
E. Atherton and R. C. Scheppard. The Peptides: analysis, synthesis, biology, Vol. 9, Academic Press, New York, 1987.
L. A. Carpino and G. Y. Han. J. Org. Chem. 37:3404–3409 (1972).
R. Knorr, A. Trzeciak, W. Bannwarth, and D. Gillessen. Tetrahedron Lett. 30:1927–1930 (1989).
L. Stamatatos, R. Leventis, N. Zuckerman, and J. R. Silvius. Biochemistry 27:3917–3925 (1988).
J. Y. Legendre and A. Supersaxo. Biochim. Biophys. Res. Commun. 217:179–185 (1995).
K. P. Ananthapadmanabhan, E. D. Goddard, N. J. Turro and P. L. Kuo. Langmuir 1:352–355 (1985).
H. Gershon, R. Ghirlando, S. B. Guttman, and A. Minsky. Biochemistry 32:7143–7151 (1993).
P. R. Gerber and K. Mueller. J. Comput. Aided Mol. Des. 9:251–268 (1995).
H. M. Berman, W. K. Olson, D. L. Beveridge, J. Westbrook, A. Gelbin, T. Demeny, S. H. Hsieh, A. R. Srinivasan, and B. Schneider. Biophys. J. 63:751–759 (1992).
N. Schmidt and J. P. Behr. Biochemistry 30:4347–4361 (1991).
W. Yang and T. A. Steitz. Cell 82:193–207 (1995).
R. P. Balasubramaniam, M. J. Bennett, A. M. Aberle, J. G. Malone, M. H. Nantz, and R. W. Malone. Gene Ther. 3:163–172 (1996).
H. Farhood, N. Serbina, and L. Huang. Biochim. Biophys. Acta 1235:289–295 (1995).
H. Ellens, J. Bentz, and F. C. Szoka. Biochemistry 23:1532–1538 (1984).
J. Y. Legendre and F. C. Szoka. Proc. Natl. Acad. Sci. USA 90:893–897 (1993).
J. H. Felgner, R. Kumar, C. N. Sridhar, C. J. Wheeler, Y. J. Tsai, R. Border, P. Ramsey, M. Martin and P. L. Felgner. J. Biol. Chem. 269:2550–2561 (1994).
E. Wagner, M. Cotten, R. Foisner and M. Birnstiel. Proc. Natl. Acad. Sci. USA 88:4255–4259 (1991).
X. Zhou and L. Huang. Biochim. Biophys. Acta 1189:195–203 (1994).
E. Wagner, D. Curiel, and M. Cotten. Adv. Drug Deliv. Rev. 14:113–135 (1994).
C. J. Chu, J. Dijkstra, M. Z. Lai, K. Hong and F. C. Szoka. Pharm. Res. 7:824–834 (1990).
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Legendre, JY., Trzeciak, A., Bur, D. et al. N-Acyl-(α,γ Diaminobutyric Acid)n Hydrazide as an Efficient Gene Transfer Vector in Mammalian Cells in Culture. Pharm Res 14, 619–624 (1997). https://doi.org/10.1023/A:1012105128722
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DOI: https://doi.org/10.1023/A:1012105128722