Improved synthesis of trinucleotide phosphoramidites and generation of randomized oligonucleotide libraries

Nucleosides Nucleotides Nucleic Acids. 2007;26(5):473-97. doi: 10.1080/15257770701426260.

Abstract

A new method to produce a set of 20 high quality trinucleotide phosphoramidites on a 5-10 g scale each was developed. The procedure starts with condensation reactions of P-components with N-acyl nucleosides, bearing the 3 '-hydroxyl function protected with 2-azidomethylbenzoyl, to give fully protected dinucleoside phosphates 13. Upon cleavage of dimethoxytrityl group from 13, dinucleoside phosphates 16 are initially transformed into trinucleoside diphosphates 19 and then the 2-azidomethylbenzoyl is selectively removed under neutral conditions to generate trinucleoside diphosphates 5 in excellent yield. Subsequent 3 '-phosphitylation affords target trinucleotide phosphoramidites 7. When mutagenic oligonucleotides are synthesized employing mixtures of building blocks 7 as well as following the new synthetic protocol, representative oligonucleotide libraries are generated in good yields.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Chromatography, High Pressure Liquid
  • Models, Chemical
  • Molecular Structure
  • Oligonucleotides / chemistry*
  • Organophosphorus Compounds / chemical synthesis*
  • Organophosphorus Compounds / chemistry
  • Spectrometry, Mass, Electrospray Ionization
  • Thionucleotides / chemistry

Substances

  • Oligonucleotides
  • Organophosphorus Compounds
  • Thionucleotides
  • phosphoramidite