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Improvement of in vivo stability of phosphodiester oligonucleotide using anionic liposomes in mice
M C De Oliveira1, V Boutet, E Fattal
1Faculdade de Farmácia da Universidade Federal de Minas Gerais, Belo Horizonte, Brazil.
Abstract:
Antisense phosphodiester oligonucleotides (ODN) are unstable in biological fluids due to nuclease-mediated degradation and therefore cannot be used in most antisense therapeutic applications. We describe here an in vitro and in vivo stabilization of a 15 mer phosphodiester sequence using anionic liposomes. Two formulations have been studied: DOPC/OA/CHOL and DOPE/OA/CHOL (pH-sensitive liposomes). Our in vitro findings reveal the same stabilization effect in mouse plasma for both anionic liposomes. In vivo investigation showed a great protective effect for both formulations after intravenous administration to mice. By contrast with in vitro results, a higher protection of ODN was observed with DOPC/OA/CHOL liposomes compared to the DOPE/OA/CHOL formulation. The latter was degraded in blood (75% of the injected dose at 5 min) probably due to interactions with blood components, and the remaining (25% at 5 min) was distributed mostly to the liver and spleen. DOPC liposomes were remarkably stable in blood and were distributed more slowly to all studied organs (liver, spleen, kidneys and lungs). Intact ODN was still observed in some organs (liver, spleen, lungs), but not in blood, 24 hours after DOPC liposome administration. These results suggest that this antisense strategy using carrier systems may be applicable to the treatment of diseases involving the reticuloendothelial system.
Insights
Anionic liposomes stabilize antisense oligonucleotides (ODN) against degradation. DOPC/OA/CHOL liposomes show superior in vivo protection, enabling potential antisense therapy applications.
Area of Science:
- Biotechnology
- Drug Delivery Systems
- Oligonucleotide Therapeutics
Background:
- Antisense phosphodiester oligonucleotides (ODN) face nuclease degradation in biological fluids, limiting their therapeutic use.
- Effective delivery and stabilization are crucial for successful antisense drug development.
Purpose of the Study:
- To evaluate the in vitro and in vivo stabilization of antisense ODN using anionic liposomes.
- To compare the efficacy of two anionic liposome formulations (DOPC/OA/CHOL and DOPE/OA/CHOL) in protecting ODN.
Main Methods:
- In vitro stability assessment of ODN in mouse plasma with anionic liposomes.
- In vivo intravenous administration of ODN-liposome formulations to mice.
- Quantification of ODN degradation and biodistribution in various organs over time.
Main Results:
- Both liposome formulations demonstrated in vitro stabilization of ODN in mouse plasma.
- In vivo, DOPC/OA/CHOL liposomes provided significantly higher ODN protection compared to DOPE/OA/CHOL.
- DOPE/OA/CHOL liposomes showed rapid ODN degradation in blood, with distribution primarily to the liver and spleen.
- DOPC/OA/CHOL liposomes exhibited remarkable stability in blood, with slower distribution to organs and intact ODN detected 24 hours post-administration.
Conclusions:
- Anionic liposomes, particularly the DOPC/OA/CHOL formulation, effectively stabilize antisense ODN in vivo.
- This liposomal delivery system shows promise for antisense therapeutic strategies, especially for diseases affecting the reticuloendothelial system.