СИНТЕЗ КОНЪЮГАТОВ ОЛИГОНУКЛЕОТИДОВ С ПРОИЗВОДНЫМИ 11,11-D2-ЛИНОЛЕВОЙ КИСЛОТЫ ПО РЕАКЦИИ АЗИД-АЛКИНОВОГО ЦИКЛОПРИСОЕДИНЕНИЯ

Предложен новый подход к синтезу конъюгатов олигонуклеотидов и жирных кислот, основанный на медь-катализируемой реакции циклоприсоединения азидсодержащих олигонуклеотидов к производным жирных кислот с терминальной тройной связью (CuAAC). Для демонстрации этого подхода осуществлен синтез пяти производных 11,11-D2-линолевой кислоты, содержащих концевую тройную связь в различных частях молекулы. Разработана методика их конъюгации с азидсодержащим модельным олигонуклеотидом Т20, проверено влияние различных условий (времени, концентрации катализатора, избытка алкинсодержащего реагента, состава растворителя и др.) на выход продукта. Такие конъюгаты устойчивы в биологических средах, обладают повышенной способностью проникать в клетки и могут найти применение в качестве средств генной терапии. 

1. Modifications in Therapeutic Oligonucleotides Improving the Delivery / I. Dovydenko [et al.]. – 2016. – P. 319–337.

2. Molecular aptamers for drug delivery / W. Tan [et al.] // Trends Biotechnol. – 2011. – Vol. 29, № 12. – P. 634–640.

3. Gene therapy clinical trials worldwide to 2012 – an update / S. L. Ginn [et al.] // J. Gene Med. – 2013. – Vol. 15, № 2. – P. 65–77.

4. Walther, W. Viral Vectors for Gene Transfer / W. Walther, U. Stein // Drugs. – 2000. – Vol. 60, № 2. – P. 249–271.

5. Thomas, C. E. Progress and problems with the use of viral vectors for gene therapy / C. E. Thomas, A. Ehrhardt, M. A. Kay // Nat. Rev. Genet. – 2003. – Vol. 4, № 5 – P. 346–358.

6. Kay, M. A. State-of-the-art gene-based therapies: the road ahead / M. A. Kay // Nat. Rev. Genet. – 2011. – Vol. 12, № 5. – P. 316–328.

7. Recent advances in characterization of nonviral vectors for delivery of nucleic acids: impact on their biological performance / C. Oliveira [et al.] // Expert Opin. Drug Deliv. – 2015. – Vol. 12, № 1. – P. 27–39.

8. Balazs, D. A. Liposomes for Use in Gene Delivery / D. A. Balazs, W. Godbey // J. Drug Deliv. – 2011. – Vol. 2011. – P. 1–12.

9. Dizaj, S. A sight on the current nanoparticle-based gene delivery vectors / S. Dizaj, S. Jafari, A. Khosroushahi // Nanoscale Res. Lett. – 2014. – Vol. 9, № 1. – P. 252.

10. Kuo, W.-T. Intracellular trafficking, metabolism and toxicity of current gene carriers. / W.-T. Kuo, H.-Y. Huang, Y.-Y. Huang // Curr. Drug Metab. – 2009. – Vol. 10, № 8. – P. 885–894.

11. Lipid Conjugated Oligonucleotides: A Useful Strategy for Delivery / M. Raouane [et al.] // Bioconjug. Chem. – 2012. – Vol. 23, № 6. – P. 1091–1104.

12. Synthesis of Oligonucleotides Carrying Amino Lipid Groups at the 3′-End for RNA Interference Studies / S. Grijalvo [et al.] // J. Org. Chem. – 2010. – Vol. 75, № 20. – P. 6806–6813.

13. Synthesis of Lipid-Oligonucleotide Conjugates for RNA Interference Studies / S. Grijalvo [et al.] // Chem. Biodivers. – 2011. – Vol. 8, № 2. – P. 287–299.

14. Synthesis, Characterization, and in Vivo Delivery of siRNA-Squalene Nanoparticles Targeting Fusion Oncogene in Papillary Thyroid Carcinoma / M. Raouane [et al.] // J. Med. Chem. – 2011. – Vol. 54, № 12. – P. 4067–4076.

15. Efficient In Vivo Delivery of siRNA to the Liver by Conjugation of α-Tocopherol / K. Nishina [et al.] // Mol. Ther. – 2008. – Vol. 16, № 4. – P. 734–740.

16. Cholesteryl-conjugated oligonucleotides: synthesis, properties, and activity as inhibitors of replication of human immunodeficiency virus in cell culture. / R.L. Letsinger [et al.] // Proc. Natl. Acad. Sci. U.S.A. – 1989. – Vol. 86, № 17.

17. Lung Delivery Studies Using siRNA Conjugated to TAT(48−60) and Penetratin Reveal Peptide Induced Reduction in Gene Expression and Induction of Innate Immunity / S.A. Moschos [et al.] // Bioconjug. Chem. – 2007. – Vol. 18, № 5. – P. 1450–1459.

18. Synthesis and silencing properties of siRNAs possessing lipophilic groups at their 3’-termini / Y. Ueno [et al.] // Nucleic Acids Symp. Ser. – 2008. – Vol. 52, № 1. – P. 503–504.

19. Habus, I. Synthesis, hybridization properties, nuclease stability, and cellular uptake of the oligonucleotide– amino-beta-cyclodextrins and adamantane conjugates. / I. Habus, Q. Zhao, S. Agrawal // Bioconjug. Chem. – Vol. 6, № 4. – P. 327–331.

20. 3’-End conjugates of minimally phosphorothioate-protected oligonucleotides with 1-O-hexadecylglycerol: synthesis and anti-ras activity in radiation-resistant cells / A. Rait [et al.] // Bioconjug. Chem. – Vol. 11, № 2. – P. 153–160.

21. Acridine- and cholesterol-derivatized solid supports for improved synthesis of 3’-modified oligonucleotides. / M. W. Reed [et al.] // Bioconjug. Chem. – Vol. 2, № 4. – P. 217–225.

22. Synthesis of cholesteryl supports and phosphoramidites containing a novel peptidyl linker for automated synthesis of triple-helix forming oligonucleotides (TFOs). / H. Vu [et al.] // Nucleic Acids Symp. Ser. – 1993, № 29. – P. 19–20.

23. Vu, H. Synthesis and Properties of Cholesteryl-Modified Triple-Helix Forming Oligonucleotides Containing a Triglycyl Linker / H. Vu, T.S. Hill, K. Jayaraman // Bioconjug. Chem. – 1994. – Vol. 5, № 6. – P. 666–668.

24. Will, D. W. Attachment of vitamin E derivatives to oligonucleotides during solid-phase synthesis / D. W. Will, T. Brown // Tetrahedron Lett. – 1992. – Vol. 33, № 19. – P. 2729–2732.

25. Synthesis and physical properties of anti-HIV antisense oligonucleotides bearing terminal lipophilic groups. / C. MacKellar [et al.] // Nucleic Acids Res. – 1992. – Vol. 20, № 13. – P. 3411–347.

26. Structure-activity relationships of cytotoxic cholesterol-modified DNA duplexes. / M. W. Reed [et al.] // J. Med. Chem. – 1995. – Vol. 38, № 22. – P. 4587–4596.

27. Matysiak, S. Acetal Oligonucleotide Conjugates in Antisense Strategy / S. Matysiak, R. Frank, W. Pfleiderer // Nucleosides and Nucleotides. – 1997. – Vol. 16, № 5–6. – P. 855–861.

28. Godeau, G. Lipid-Conjugated Oligonucleotides via “Click Chemistry” Efficiently Inhibit Hepatitis C Virus Translation / G. Godeau, C. Staedel, P. Barthélémy // J. Med. Chem. – 2008. – Vol. 51, № 15. – P. 4374–4376.

29. Azide Phosphoramidite in Direct Synthesis of Azide-Modified Oligonucleotides / M. A. Fomich [et al.] // Org. Lett. – 2014. – Vol. 16, № 17. – P. 4590–4593.

30. Isotope-reinforced polyunsaturated fatty acids protect mitochondria from oxidative stress / A. Y. Andreyev [et al.] // Free Radic. Biol. Med. – 2015. – Vol. 82. – P. 63–72.

31. Small amounts of isotope-reinforced polyunsaturated fatty acids suppress lipid autoxidation / S. Hill [et al.] // Free Radic. Biol. Med. – 2012. – Vol. 53, № 4. – P. 893–906.

32. Dussault, P. H. Total Synthesis of the Alkoxydioxines (+)- and (−)-Chondrillin and (+)- and (−)-Plakorin via Singlet Oxygenation/Radical Rearrangement / P. H. Dussault, C. T. Eary, K. R. Woller // J. Org. Chem. – 1999. – Vol. 64, № 6. – P. 1789–1797.