Abstract
Adsorption of oligothymidylates on polyisobutyl- or polyisohexylcyanoacrylate nanoparticles was achieved in the presence of hydrophobic cations such as tetraphenylphosphonium chloride or quaternary ammonium salts. Results suggested that oligonucleotide adsorption on the nanoparticles was mediated by the formation of ion pairs between the negatively charged phosphate groups of the nucleic acid chain and the hydrophobic cations. The adsorption efficiency of oligonucleotide–cation complexes on nanoparticles was found to be highly dependent upon several parameters: oligonucleotide chain length, nature of the cyanoacrylic monomer, hydrophobicity of cations used as ion-pairing agents, and ionic concentration of the medium. Carrier capacity of polyisohexylcyanoacrylate nanoparticles for oligothymidylates (16 nucleotides) complexed with cetyltrimetylammonium bromide in the presence of 0.15 M NaCl was determined to be 5 µmol/g polymer. The in vitro protection of oligothymidylates adsorbed to nanoparticles against degradation by a 3′-exonuclease (snake venom phosphodiesterase) was also demonstrated. These results showed that nanoparticles can be considered as convenient carriers for the protection and delivery of Oligonucleotides to cells in culture and for future applications in vivo.
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REFERENCES
C. Hélène and J. J. Toulmé. Specific regulation of gene expressio by antisense, sense and antigene nucleic acids. Biochim. Biophys. Acta 1049:99–125 (1990).
E. Uhlmann and A. Peyman. Antisense oligonucleotides: A new therapeutic principle. Chem. Rev. 90:543–584 (1990).
E. Wickstrom. Oligonucleotide stability in subcellular extracts and culture media. J. Biochem. Biophys. Methods 13:97–102 (1986).
C. Cazenave, M. Chevrier, N. T. Thuong, and C. Hélène. Rate of [α]-and [β]-oligonucleotides in Xenopus oocytes. Implications for anti-messenger strategies. Nucleic Acids Res. 15:10507–10521 (1987).
J. M. Dagle, J. A. Walder, and D. L. Weeks. Targeted degradation of mRNA in Xenopus oocytes and embryos directed by modified oligonucleotides: Studies of An2 and cyclin in embryogenesis. Nucleic Acids Res. 18:4751–4757 (1990).
S. L. Loke, C. A. Stein, X. H. Zhang, K. Mori, M. Nakanishi, C. Subasinghe, J. S. Cohen, and L. M. Neckers. Characterization of oligonucleotide transport into living cell. Proc. Natl. Acad. Sci. USA 86:3474–3478 (1989).
G. Zon. Oligonucleotide analogues as potential chemotherapeutic agents. Pharm. Res. 5:539–549 (1988).
P. Verspieren, A. W. C. A. Cornelissen, N. T. Thuong, C. Hélène, and J. J. Toulmé. An acridine-linked oligodeoxynucleotide targeted to the common 5′ end of trypanosome mRNAs kills cultured parasites. Gene 61:307–315 (1987).
A. S. Boutorin, L. V. Gusvkova, E. M. Ivanova, N. D. Kobetz, V. F. Zarytova, A. S. Ryte, L. V. Yurchenko, and V. V. Vlassov. Synthesis of alkylating oligonucleotides derivatives containing cholesterol or phenazinium residue at the 3′-termini and their interaction with DNA within mammalian cells. FEBS Lett. 254:129–132 (1989).
J. P. Leonetti, G. Degols, and B. Lebleu. Biological activity of oligonucleotide-poly(L-lysine) conjugates: Mechanism of cell uptake. Bioconjugate Chem. 1:149–153 (1990).
C. A. Stein, C. Subasinghe, K. Shinozuka, and J. S. Cohen. Physicochemical properties of phosphorothioate oligonucleotides. Nucleic Acids. Res. 16:3209–3221 (1988).
R. S. Quartin, C. L. Brakel, and J. G. Wetmur. Number and distribution of methylphosphonates linkages in oligodeoxynucleotides affect exo-and endonuclease sensitivity and ability to form RNase H substrates. Nucleic Acids Res. 17:7253–7262 (1989).
T. A. Bacon, F. Morvan, B. Rayner, J. L. Imbach, and E. Wickstrom. α-Oligonucleotide stability in serum, subcellular extracts and culture media. J. Biochem. Biophys. Methods 16:311–318 (1988).
S. L. Loke, C. Stein, X. Zhang, M. Avigan, J. Cohen, and L. M. Neckers. Delivery of c-myc antisense phosphorothioate oligonucleotides to hematopoietic cells in culture by liposomes fusion: Specific reduction in c-myc protein expression correlates with inhibition of cell growth and DNA synthesis. Current Top. Microbiol. Immunol. 141:282–289 (1988).
J. P. Leonetti, P. Machy, G. Degols, B. Lebleu, and L. Leserman. Antibody-targeted liposomes containing oligodeoxyribo-nucleotides complementary to viral RNA selectively inhibit viral replication. Proc. Natl. Acad. Sci. USA 87:2448–2451 (1990).
P. Couvreur, B. Kante, M. Roland, P. Guiot, P. Baudhuin, and P. Speiser. Polycyanoacrylate nanocapsules as potential lysosomotropic carriers: Preparation, morphological and sorptive properties. J. Pharm. Pharmacol. 31:331–332 (1979).
V. Guise, P. Jaffray, J. Delattre, F. Puisieux, M. Adolphe, and P. Couvreur. Comparative cell uptake of propidium iodide associated with liposomes or nanoparticles. Cell. Mol. Biol. 33:397–405 (1987).
P. Guiot and P. Couvreur. Quantitative study of the interaction between polybutylcyanoacrylate nanoparticles and mouse peritoneal macrophages in culture. J. Pharm. Belg. 38:130–134 (1983).
P. Couvreur, L. Grislain, V. Lenaerts, F. Brasseur, P. Guiot, and A. Biernacki. Biodegradable polymeric nanoparticles as drug carrier for antitumor agents. In Guiot, P., and Couvreur, P., (eds.), Polymeric Nanoparticles and Microspheres, CRC Press, Boca Raton, FL, 1986, pp. 27–94.
N. Chiannilkulchai, Z. Driouich, J. P. Benoit, A. L. Parodi, and P. Couvreur. Doxorubicin-loaded nanoparticles: Increased efficiency in murine hepatic metastases. Select. Cancer Ther. 5:1–11 (1989).
E. Fattal, M. Youssef, P. Couvreur, and A. Andremont. Treatment of experimental salmonellosis in mice with ampicillin-bound nanoparticles. Antimicrob. Agents Chemother. 33:1540–1543 (1989).
V. Guise, J. Y. Drouin, J. Benoit, J. Mahuteau, P. Dumont, and P. Couvreur. Vidarabine-loaded nanoparticles: A physicochemical study. Pharm. Res. 7:736–741 (1990).
L. Illum, M. A. Khan, E. Mak, and S. S. Davis. Evaluation of carrier capacity and release characteristics for poly (butyl 2-cyanoacrylate) nanoparticles. Int. J. Pharm. 30:17–28 (1986).
L. Illum, I. M. Hunneyball, and S. S. Davis. The effect of hydrophilic coatings on the uptake of colloidal particles by the liver and by peritoneal macrophages. Int. J. Pharm. 29:53–65 (1986).
S. J. Douglas, L. Illum, S. S. Davis, and J. Kreuter. Particle size and size distribution of poly(butyl-2-cyanoacrylate) nanoparticles. I. Influence of physicochemical factors. J. Colloid Interface Sci. 101:149–158 (1984).
R. H. Müller, C. Lherm, J. Herbort, and P. Couvreur. Alkylcyanoacrylate drug carriers. I. Physicochemical characterization of nanoparticles with different alkyl chain length (submitted for publication).
P. Connor and R. H. Ottewill. The adsorption of cationic surface active agents on polystyrene surfaces. J. Colloid Interface Sci. 37:642–651 (1971).
J. Goodchild, B. Kim, and P. C. Zamecnik. The clearance and degradation of oligonucleotides following intravenous injection into rabbits. Antisense Res. Dev. 1:153–160 (1991).
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Chavany, C., Doan, T.L., Couvreur, P. et al. Polyalkylcyanoacrylate Nanoparticles as Polymeric Carriers for Antisense Oligonucleotides. Pharm Res 9, 441–449 (1992). https://doi.org/10.1023/A:1015871809313
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DOI: https://doi.org/10.1023/A:1015871809313