New non nucleosidic phosphoramidite reagent for solid phase synthesis of biotinylated oligonucleotides _net

Bioorganic and Medicinal Chemistry Letters

Volumn 6, Issue 2, 1996, Pages 147-152

  Neuner, Philippe ^a  

^a IRBM   (Italy)

Author keywords

[No Author keywords available]

Indexed keywords

BIOTIN DERIVATIVE; OLIGONUCLEOTIDE;

ARTICLE; DNA SYNTHESIS; DRUG SYNTHESIS; HIGH PERFORMANCE LIQUID CHROMATOGRAPHY; REACTION ANALYSIS; TECHNIQUE;

EID: 0030039001     PISSN: 0960894X     EISSN: None     Source Type: Journal    
DOI: 10.1016/0960-894X(95)00579-I     Document Type: Article

References (21)

1. De Vos, M-J., Cravador, A., Lenders, J-P., Houard, S. and Bollen, A. 1990, Nucleosides & Nucleotides, 9, 259-273.
2. Bengstrom, M., Harju, L. and Syvanen, A-C. 1991, Nucleosides & Nucleotides, 10, 507-509.
3. Reyes-Engel, A. and Dieguez-Lucena, J., L. 1993, Nucleic Acid Res., 21, 759-760.
4. Niemeyer, C., M., Sano, T., Smith, C., L. and Cantor, C., R. 1994, Nucleic Acid Res., 22, 5530-5539.
5. Pon, R.T., 1991, Tetrahedron Lett., 32, 1715-1718.
6. Fontanel, M-L., Bazin, H. and Teoule, R. 1993, Analytical Biochemistry, 214, 338-340.
7. 4/LiCl mixture in MeOH/THF 1/1. The reaction mixture was diluted with dichloromethane and washed first with a 10% aqueous citric acid solution, then with a 1% aqueous sodium bicarbonate solution. The organic layer was separated, dried over sodium sulfate and evaporated to dryness. The resulting oil was purified by flash chromatography (FC) using a gradient of 3-6% methanol in dichloromethane as eluent. The pure product was obtained in 73% yield. While this manuscript was in preparation, De Vos et al. reported the synthesis of 4-amino-1,3-butanediol through a different approach. De Vos, M.J., Van Elsen, A. and Bollen, A. 1994, Nucleosides & Nucleotides, 13, 2245-2265.
8. We found it necessary to protect the secondary alcohol in 2 to avoid side reaction during the preparation of 4 using the mixed anhydride methodology.
9. The preparation of the Fmoc protected amino hexanoic acid is described by Haralambis J., Duncan L., Angus K. and Tregear G. W. 1990, Nucleic Acid Res., 18, 493-499.
10. 1 eq. of N-methylmorpholine was added to a stirring solution of 1 eq. of 6-N-Fmoc-amino hexanoic acid in dry THF at -30°C under anhydrous conditions. After 15 min, 1 eq. of isobutyl chloroformate was added. The cloudy mixture was stirred at -30°C for 30 min, then at 0°C for 30 min. Tlc (2% MeOH/DCM) showed a new spot Rf 0.80 corresponding to the mixed anhydride. The stirring mixture was cooled to -30°C and the previously amino deprotected 3 in THF was added in one portion. Reaction was completed in 1.50 h.
11. 2 Si: C, 72.85; H, 7.52; N, 3.27. Found: C, 72.30; H, 7.85; N, 3.22.
12. Parameswaran, K.N. 1990, Org. Prep. Proced. Int., 22, 119-121. The pure compound 5 was obtained in 75 % yield after flash chromatography (FC) on silica using a gradient of 6-10% methanol in dichloromethane. Rf : 0.30 in 10% methanol/dichloromethane.
13. Pieles, U., Sproat, B.S. and Lamm, G.M. 1990, Nucleic Acids Res., 18, 4355-4360. Compound 5 was obtained in 80% yield after FC using the same eluent system described in reference 12.
14. 4 S: C, 70.99; H, 6.87; N, 5.34. Found: C, 69.51; H, 7.47; N, 5.55.
15. 3) 6: 148.26, 148.15, 147.18, 147.00.
16. The purification was carried out using a gradient of buffer A (0.1M TEAA pH 7.50) and buffer B (30% buffer A/70% acetonitrile) at a flow rate of 2 ml/min. over 50 min. The purified tritylated oligomer was deblocked with 0.5 ml acetic acid/water (8/2), and after 30 min at room temperature the mixture was extracted with ether (3 × 0.5ml) and evaporated to dryness.
17. Maniatis, T., Fritsch, E.F., Sambrook, J. 1989, Molecular Cloning; Cold Spring Harbor Laboratory, New York.
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21. Li, W-B., Gruber C.E., Noon M.C., Polayes D., Schmidt B. and Jesse J. 1995, Focus, 17, 35-39.