A novel phosphitylating reagent for in situ generation of deoxyribonucleoside phosphoramidites

Tetrahedron Letters

Volumn 37, Issue 3, 1996, Pages 331-334

  Zhang, Zhaoda ^a   Tang, Jin Yan ^a  

^a HYBRIDON INC   (United States)

Author keywords

[No Author keywords available]

Indexed keywords

PHOSPHORAMIDIC ACID DERIVATIVE; RIBONUCLEOSIDE DERIVATIVE;

ARTICLE; DRUG SYNTHESIS; GEL ELECTROPHORESIS; NUCLEAR MAGNETIC RESONANCE; REACTION ANALYSIS; TECHNIQUE;

EID: 0030068363     PISSN: 00404039     EISSN: None     Source Type: Journal    
DOI: 10.1016/0040-4039(95)02165-5     Document Type: Article

References (16)

1. For reviews see: (a) Nucleic Acids in Chemistry and Biology, Blackburn, G. M.; Gait, M. J., Eds.; IRL:Oxford, 1990.
2. (b) Uhlmann, E.; Peyman, A. Chem. Rev. 1990, 90, 543 and the literatures cited therein.
3. (a) Caruthers, M H. Acc. Chem. Res. 1991, 24, 278.
4. (b) Beaucage, S. L.; Iyer, R P. Tetrahedron 1992, 48, 2223 and the literatures cited therein.
5. Sinha, N. D.; Biernat, J.; Koster, H. Tetrahedron Lett. 1983, 24, 5843
6. (a) Bodepudi, V.; Shibutani, S.; Johnson, F Chem. Res. Toxicol. 1992, 5, 608. In this paper, the authors reported a 'one pot' procedure for the synthesis of the phosphoramidite.
7. (b) Tan, W.; Iyer, R. P.; Jiang, Z.; Yu, D.; Agrawal, S. Tetrahedron Lett. 1995, 36, 5323.
8. (a) Beaucage, S. L Tetrahedron Lett. 1984, 25, 375.
9. (b) Moore, M. F.; Beaucage, S. L. J. Org. Chem. 1985, 50, 2019.
10. Barone, A. D.; Tang, J.-Y., Caruthers, M. H. Nucleic Acids Res., 1984, 12, 4051.
11. 3) δ 133.9.
12. 3) 6 138.2.
13. 3CN (10.0 mL) at room temperature. The mixture was stirred for 10 min, and the phosphoramidite solution (0.1 M) was ready to use. There was no precipitate found in the phosphoramidite solution prepared in situ using 1. (b) In a similar manner, we carried out the synthesis using 1, 5'-DMT-thymidine and diisopropylethylamine in acetonitrile After the solvent was removed, the reaction mixture was purified by silica-gel chromatography to give 5'-DMT-thymidine phosphoramidite in 94% yield.
14. The oligonucleotides were synthesized on a 0.2 or 1 μmol scale following the standard protocol in using an automated synthesizer (Millipore 8909 Expedite™, Bedford, MA). Eight-hour treatment with ammonium hydroxide at 65 °C was carried out to cleave the oligomer from the support and to deprotect nucleoside bases. The mixture was then filtered to remove the CPG. After the ammonium hydroxide solution was removed by Speed Vac, the remaining crude products were submitted for CE and IEX-HPLC analysis.
15. (a) An antisense oligonucleotide inhibitor of HIV-1 replication developed by Hybridon, Inc., (b) Agrawal, S., Tang, J. Y. Antisense Res. Dev. 1992, 4, 261.
16. Sanghvi, Y. S. In Antisense Research and Applications, Crook, S. T.; Lebleu, B., Eds., CRC: Ann Arbor, 1993, 273, and the literatures cited therein.