Skip to main content

Advertisement

SpringerLink
Log in

Bioresponsive Deciduous-Charge Amphiphiles for Liposomal Delivery of DNA and siRNA

  • Research Paper
  • Published:
Pharmaceutical Research Aims and scope Submit manuscript

ABSTRACT

Purpose

Biolabile cationic lipids were developed for efficient intracellular delivery of DNA and siRNA.

Methods

The compounds have been designed starting from the membrane lipid DOPC in a way they may loose their cationic charge when exposed to an acidic and/or enzymatic stimulus, such as those met during the journey of a lipoplex in biological media.

Results

They demonstrated remarkable efficiency to deliver DNA in various cell lines (BHK-21, Calu-3, NCI-H292, and A549), with no significant cytotoxicity. Furthermore, two of the compounds (carbonate-based DOPC derivatives) revealed able to deliver small interfering RNA in U87Luc and A549Luc cancer cells and to mediate a selective 70–80% knockdown of the stably transfected luciferase gene.

Conclusions

The results show that the described bioresponsive cationic lipids have high DNA and siARN delivery activity which is encouraging in view of delivering a therapeutic nucleic acid to pulmonary tissues in vivo.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Scheme 1
Scheme 2
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7
Fig. 8
Fig. 9

Similar content being viewed by others

REFERENCES

  1. Behr JP. DNA strongly binds to micelles and vesicles containing lipopolyamines or lipointercalants. Tetrahedron Lett. 1986;27:5861–4.

    Article  CAS  Google Scholar 

  2. Felgner PL, Gadek TR, Holm M, Roman R, Chan HW, Wenz M, Noethrop JP, Ringold GM, Danielsen M. Lipofection: a highly efficient, lipid-mediated DNA-transfection procedure. Proc Natl Acad Sci U S A. 1987;84:7413–7.

    Article  PubMed  CAS  Google Scholar 

  3. Felgner JH, Kumar R, Sridhar CN, Wheeler CJ, Tsai YJ, Border R, Ramsey P, Martin M, Felgner PL. Enhanced gene delivery and mechanism studies with a novel series of cationic lipid formulations. J Biol Chem. 1994;269:2550–61.

    PubMed  CAS  Google Scholar 

  4. Godbey WT, Wu KK, Mikos A. Tracking the intracellular path of poly(ethylenimine)/DNA complexes for gene delivery. Proc Natl Acad Sci U S A. 1999;96:5177–81.

    Article  PubMed  CAS  Google Scholar 

  5. Lu JJ, Langer R, Chen J. A novel mechanism is involved in cationic lipid-mediated functional siRNA delivery. Mol Pharm. 2009;6:763–71.

    Article  PubMed  CAS  Google Scholar 

  6. Ming X, Sato K, Juliano RL. Unconventional internalization mechanisms underlying functional delivery of antisense oligonucleotides via cationic lipoplexes and polyplexes. J Control Release. 2011;153:83–92.

    Article  PubMed  CAS  Google Scholar 

  7. Boussif O, Lezoualc’h F, Zanta MA, Mergny MD, Scherman D, Demeneix B, Behr JP. A versatile vector for gene and oligonucleotide transfer into cells in culture and in vivo: polyethylenimine. Proc Natl Acad Sci U S A. 1995;92:7297–301.

    Article  PubMed  CAS  Google Scholar 

  8. Koynova R, Wang L, MacDonald RC. An intracellular lamellar-nonlamellar phase transition rationalizes the superior performance of some cationic lipid transfection agents. Proc Natl Acad Sci U S A. 2006;103:14373–8.

    Article  PubMed  CAS  Google Scholar 

  9. Wagner E, Plank C, Zatloukal K, Cotten M, Birnstiel ML. Influenza virus hemagglutinin HA-2 N-terminal fusogenic peptides augment gene transfer by transferrin-polylysine-DNA complexes: toward a synthetic virus-like gene-transfer vehicle. Proc Natl Acad Sci U S A. 1992;89:7934–8.

    Article  PubMed  CAS  Google Scholar 

  10. Capecchi MR. High efficiency transformation by direct microinjection of DNA into cultured mammalian cells. Cell. 1980;22:479–88.

    Article  PubMed  CAS  Google Scholar 

  11. Pozharski E, MacDonald RC. Lipoplex thermodynamics: determination of DNA-cationic lipoid interaction energies. Biophys J. 2003;85:3969–78.

    Article  PubMed  CAS  Google Scholar 

  12. Xu XH, Szoka FC. Mechanism of DNA release from cationic liposome/DNA complexes used in cell transfection. Biochemistry. 1996;35:5616–23.

    Article  PubMed  CAS  Google Scholar 

  13. Prata CAH, Zhao YX, Barthelemy P, Li YG, Luo D, McIntosh TJ, Lee SJ, Grinstaff MW. Charge-reversal amphiphles for gene delivery. J Am Chem Soc. 2004;126:12196–7.

    Article  PubMed  CAS  Google Scholar 

  14. Zhang XX, Prata CAH, McIntosh TJ, Barthelemy P, Grinstaff MW. The effect of charge-reversal amphiphile spacer composition on DNA and siRNA delivery. Bioconjugate Chem. 2010;21:988–93.

    Article  CAS  Google Scholar 

  15. Zhang XX, Allen PG, Grinstaff MW. Macropinocytosis is the major pathway responsible for DNA transfection in CHO cells by a charge-reversal amphiphile. Mol Pharm. 2011;8:758–66.

    Article  PubMed  CAS  Google Scholar 

  16. MacDonald RC, Ashley GW, Shida MM, Rakhmanova VA, Tarahovsky YS, Pantazatos DP, Kennedy MT, Pozharski EV, Baker KA, Jones RD, Rosenzweig HS, Choi KL, Qiu RZ, McIntosh TJ. Physical and biological properties of cationic triesters of phosphatidylcholine. Biophys J. 1999;77:2612–29.

    Article  PubMed  CAS  Google Scholar 

  17. MacDonald RC, Rakhmanova VA, Choi KL, Rosenzweig HS, Lahiri MK. O-Ethylphosphatidylcholine: a metabolizable cationic phospholipid which is a serum-compatible DNA transfection agent. J Pharm Sci. 1999;88:896–904.

    Article  PubMed  CAS  Google Scholar 

  18. Pierrat P, Creusat G, Laverny G, Pons F, Zuber G, Lebeau L. A Cationic phospholipid–detergent conjugate as a new efficient carrier for siRNA delivery. Chem Eur J. 2012;18:3835–9.

    Article  PubMed  CAS  Google Scholar 

  19. Pierrat P, Laverny G, Creusat G, Wehrung P, Strub JM, Van Dorsselaer A, et al. Phospholipid-detergent conjugates as efficient lipidic carriers for siRNA delivery. Chem Eur J. in press, doi:10.1002/chem.201203071.

  20. Lebeau L, Olland S, Oudet P, Mioskowski C. Rational design and synthesis of phospholipids for the two-dimensional crystallization of DNA gyrase, a key element in chromosome organization. Chem Phys Lipids. 1992;62:93–103.

    Article  PubMed  CAS  Google Scholar 

  21. Zanta MA, Boussif O, Adib A, Behr JP. In vitro gene delivery to hepatocytes with galactosylated polyethylenimine. Bioconjugate Chem. 1997;8:839–44.

    Article  CAS  Google Scholar 

  22. Foillard S, Zuber G, Doris E. Polyethylenimine-carbon nanotube nanohybrids for siRNA-mediated gene silencing at cellular level. Nanoscale. 2011;3:1461–4.

    Article  PubMed  CAS  Google Scholar 

  23. Dikmen ZG, Gellert GC, Jackson S, Gryaznov S, Tressler R, Dogan P, Wright WE, Shay JW. In vivo inhibition of lung cancer by GRN163L: a novel human telomerase inhibitor. Cancer Res. 2005;65:7866–73.

    PubMed  CAS  Google Scholar 

  24. Gentine P, Bubel A, Crucifix C, Bourel-Bonnet L, Frisch B. Manufacture of liposomes by isopropanol injection: characterization of the method. J Liposome Res. 2011;22:18–30.

    Article  PubMed  Google Scholar 

  25. Gorman CM, Aikawa M, Fox B, Fox E, Lapuz C, Michaud B, Nguyen H, Roche E, Sawa T, WienerKronish JP. Efficient in vivo delivery of DNA to pulmonary cells using the novel lipid EDMPC. Gene Ther. 1997;4:983–92.

    Article  PubMed  CAS  Google Scholar 

  26. Shpirt AM, Kononov LO, Maltsev SD, Shibaev VN. Chemical synthesis of polyprenyl sialyl phosphate, a probable biosynthetic intermediate of bacterial polysialic acid. Carbohydr Res. 2011;346:2849–54.

    PubMed  CAS  Google Scholar 

  27. Farquhar D, Khan S, Wilkerson MC, Andersson BS. Biologically-cleavable phosphate protective groups: 4-acyloxy-1,3,2-dioxaphosphorinanes as neutral latent precursors of dianionic phosphates. Tetrahedron Lett. 1995;36:655–8.

    Article  CAS  Google Scholar 

  28. Crich D, Picard S. Highly stereoselective synthesis of α-D-mannopyranosyl phosphosugars. J Org Chem. 2009;74:9576–9.

    Article  PubMed  CAS  Google Scholar 

  29. Yang Y, Babiak P, Reymond JL. New monofunctionalized fluorescein derivatives for the efficient high-throughput screening of lipases and esterases in aqueous media. Helv Chim Acta. 2006;89:404–15.

    Article  CAS  Google Scholar 

  30. Thomas JD, Sloan KB. Overcoming steric effects in the coupling reaction of alkyloxycarbonyloxymethyl (AOCOM) halides with phenols: an efficient synthesis of AOCOM phenolic prodrugs. Tetrahedron Lett. 2007;48:109–12.

    Article  CAS  Google Scholar 

  31. Ostergaard J, Larsen C. Bioreversible derivatives of phenol. 2. Reactivity of carbonate esters with fatty acid-like structures towards hydrolysis in aqueous solutions. Molecules. 2007;12:2396–412.

    Article  PubMed  CAS  Google Scholar 

  32. Ross PC, Hui SW. Lipoplex size is a major determinant of in vitro lipofection efficiency. Gene Ther. 1999;6:651–9.

    Article  PubMed  CAS  Google Scholar 

  33. Hübner W, Blume A. Interactions at the lipid–water interface. Chem Phys Lipids. 1998;96:99–123.

    Article  Google Scholar 

  34. Geall AJ, Blagbrough IS. Rapid and sensitive ethidium bromide fluorescence quenching assay of polyamine conjugate—DNA interactions for the analysis of lipoplex formation in gene therapy. J Pharm Biomed Anal. 2000;22:849–59.

    Article  PubMed  CAS  Google Scholar 

  35. Rosenzweig HS, Rakhmanova VA, McIntosh TJ, MacDonald RC. O-Alkyl dioleoylphosphatidylcholinium compounds: the effect of varying alkyl chain length on their physical properties and in vitro DNA transfection activity. Bioconjugate Chem. 2000;11:306–13.

    Article  CAS  Google Scholar 

  36. Wang L, Koynova R, Parikh H, MacDonald RC. Transfection activity of binary mixtures of cationic O-substituted phosphatidylcholine derivatives: the hydrophobic core strongly modulates physical properties and DNA delivery efficacy. Biophys J. 2006;91:3692–706.

    Article  PubMed  CAS  Google Scholar 

  37. Munnecke DM. Enzymatic-hydrolysis of organophosphate insecticides, a possible pesticide disposal method. Appl Environ Microbiol. 1976;32:7–13.

    PubMed  CAS  Google Scholar 

  38. Heeg K, Reimann J, Kabelitz D, Hardt C, Wagner H. A rapid colorimetric assay for the determination of IL-2-producing Helper T-cell frequencies. J Immunol Methods. 1985;77:237–46.

    Article  PubMed  CAS  Google Scholar 

  39. Hui SW, Langner M, Zhao YL, Ross P, Hurley E, Chan K. The role of helper lipids in cationic liposome-mediated gene transfer. Biophys J. 1996;71:590–9.

    Article  PubMed  CAS  Google Scholar 

  40. Koltover I, Salditt T, Radler JO, Safinya CR. An inverted hexagonal phase of cationic liposome-DNA complexes related to DNA release and delivery. Science. 1998;281:78–81.

    Article  PubMed  CAS  Google Scholar 

  41. Sanders N, Rudolph C, Braeckmans K, De Smedt SC, Demeester J. Extracellular barriers in respiratory gene therapy. Adv Drug Deliv Rev. 2009;61:115–27.

    Article  PubMed  CAS  Google Scholar 

  42. Rosenecker J, Naundorf S, Gersting SW, Hauck RW, Gessner A, Nicklaus P, Müller RH, Rudolph C. Interaction of bronchoalveolar lavage fluid with polyplexes and lipoplexes: analysing the role of proteins and glycoproteins. J Gene Med. 2003;5:49–60.

    Article  PubMed  CAS  Google Scholar 

  43. Merkel OM, Beyerle A, Librizzi D, Pfestroff A, Behr TM, Sproat B, Barth PJ, Kissel T. Nonviral siRNA delivery to the lung: Investigation of PEG-PEI polyplexes and their in vivo performance. Mol Pharm. 2009;6:1246–60.

    Article  PubMed  CAS  Google Scholar 

Download references

ACKNOWLEDGMENTS AND DISCLOSURES

This work was supported by the Agence Nationale pour la Recherche (ANR, France).

Author information

Authors and Affiliations

  1. Laboratoire de Conception et Application de Molécules Bioactives Faculté de Pharmacie, CNRS - Université de Strasbourg, 74 route du Rhin, BP 60024, 67401, Illkirch, France

    Philippe Pierrat, Dimitri Kereselidze, Patrick Wehrung, Guy Zuber, Françoise Pons & Luc Lebeau

Authors
  1. Philippe Pierrat
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Dimitri Kereselidze
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Patrick Wehrung
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Guy Zuber
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Françoise Pons
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Luc Lebeau
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Luc Lebeau.

Electronic supplementary material

Below is the link to the electronic supplementary material.

ESM 1

(DOCX 26 mb)

Rights and permissions

Reprints and permissions

About this article

Cite this article

Pierrat, P., Kereselidze, D., Wehrung, P. et al. Bioresponsive Deciduous-Charge Amphiphiles for Liposomal Delivery of DNA and siRNA. Pharm Res 30, 1362–1379 (2013). https://doi.org/10.1007/s11095-013-0976-9

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11095-013-0976-9

KEY WORDS

Search

Navigation