Synthesis of 2′‐O‐β‐d‐Ribofuranosylnucleosides

Sergey N. Mikhailov1, Ekaterina V. Efimtseva1, Andrei A. Rodionov1, Georgii V. Bobkov1, Irina V. Kulikova1, Piet Herdewijn2

1 Russian Academy of Sciences, Engelhardt Institute of Molecular Biology, Moscow, Russia, 2 Rega Institute for Medical Research, Leuven, Belgium
Publication Name:  Current Protocols in Nucleic Acid Chemistry
Unit Number:  Unit 1.14
DOI:  10.1002/0471142700.nc0114s27
Online Posting Date:  January, 2007
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Abstract

A simple and efficient method for the preparation of 2‐O‐β‐D‐ribofuranosylnucleosides, minor tRNA components, is described in this unit. The method consists of condensation of a small excess of 1‐O‐acetyl‐2,3,5‐tri‐O‐benzoyl‐β‐D‐ribofuranose activated with tin tetrachloride with N‐protected 3,5‐O‐tetra‐isopropyldisiloxane‐1,3‐diyl‐ribonucleosides in 1,2‐dichloroethane. Subsequent deprotection produces 2‐O‐β‐D‐ribofuranosylnucleosides in an overall yield of 46% to 72%.

Keywords: disaccharide nucleosides; minor tRNA components; synthesis

     
 
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Table of Contents

  • Basic Protocol 1: Preparation of 2′‐O‐β‐D‐Ribofuranosyladenosine
  • Alternate Protocol 1: Preparation of 2′‐O‐β‐D‐Ribofuranosyluridine
  • Alternate Protocol 2: Preparation of 2′‐O‐β‐D‐Ribofuranosylthymidine
  • Alternate Protocol 3: Preparation of 2′‐O‐β‐D‐Ribofuranosylcytidine
  • Alternate Protocol 4: Preparation of 2′‐O‐β‐D‐Ribofuranosylguanosine
  • Support Protocol 1: Preparation of the 3′,5′‐Protected Ribothymidine
  • Commentary
  • Literature Cited
  • Figures
     
 
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Materials

Basic Protocol 1: Preparation of 2′‐O‐β‐D‐Ribofuranosyladenosine

  Materials
  • 1‐O‐Acetyl‐2,3,5‐tri‐O‐benzoyl‐β‐D‐ribofuranose (S.2)
  • N6‐Benzoyl‐3′,5′‐(1,1,3,3‐O‐tetraisopropyldisiloxane‐1,3‐diyl)adenosine (S.1; unit 2.4; Fig. )
  • Phosphorus pentoxide (P 2O 5)
  • Balloon of nitrogen or argon
  • 1,2‐Dichloroethane, anhydrous
  • Tin tetrachloride (SnCl 4)
  • Methanol (MeOH), analytical grade
  • Methylene chloride (CH 2Cl 2), reagent grade
  • Saturated sodium bicarbonate solution (sat. NaHCO 3)
  • Hyflo Super Cel (Fluka)
  • Sodium sulfate, anhydrous (Na 2SO 4)
  • Silica gel (e.g., Kieselgel 60, 0.06 to 0.20 mm; Merck)
  • Tetrabutylammonium fluoride trihydrate (TBAF)
  • Tetrahydrofuran (THF), reagent grade
  • Chloroform (CHCl 3), reagent grade
  • 5 M ammonia in methanol (half‐saturated at 0°C)
  • Diethyl ether, reagent grade
  • 50‐ and 250‐mL round‐bottom flasks
  • Vacuum desiccator
  • Vacuum oil pump
  • TLC plate: silica‐coated aluminum plate with fluorescent indicator (Merck silica gel 60 F 254)
  • 254‐nm UV lamp
  • Long disposable capillaries
  • 100‐mL funnels with sintered glass disc filters (porosity 3)
  • 100‐ and 250‐mL separatory funnel
  • Rotary evaporator equipped with a water aspirator
  • 3 × 20–cm and 3 × 15–cm sintered glass chromatography columns, porosity 3
  • Stainless steel spatula
  • Additional reagents and equipment for TLC ( appendix 3D) and column chromatography ( appendix 3E)

Alternate Protocol 1: Preparation of 2′‐O‐β‐D‐Ribofuranosyluridine

  • Thymine
  • Ammonium sulfate [(NH 4) 2SO 4]
  • 1,1,1,3,3,3‐Hexamethyldisilazane, reagent grade
  • Calcium chloride (CaCl 2), anhydrous
  • Trimethylsilyl trifluoromethanesulfonate (TMSOTf)
  • 0.2 N sodium methoxide (NaOMe), freshly prepared from sodium and dry methanol
  • Dowex 50 × 4 (100 to 200 mesh) in H+ form
  • Pyridine, anhydrous
  • Markiewicz reagent: 1,3‐dichloro‐1,1,3,3‐tetraisopropyldisiloxane, 96% pure (Wacker)
  • Toluene, reagent grade
  • 100‐, 250‐, and 500‐mL round‐bottom flasks
  • Reflux condensers
  • CaCl 2 protection tubes
  • Oil bath with temperature control
  • Adapters with stopcocks and vacuum pump (Fig. )
  • 250‐ and 500‐mL separatory funnels
  • Glass filters (porosity 3)
  • 3 × 20–cm chromatography columnsx
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Figures

Videos

Literature Cited

Literature Cited
   Andreeva, O.I., Golubeva, A.S., Kochetkov, S.N., Van Aerschot, A., Herdewijn, P., Efimtseva, E.V., Ermolinsky, B.S., and Mikhailov, S.N. 2002. An additional 2′‐ribofuranose residue at a specific position of DNA primer prevents its elongation by HIV‐1 reverse trancriptase. Bioorg. Med. Chem. Lett. 12:681‐684.
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