Synthesis of phosphorothioamidites derived from 3'-thio-3'-deoxythymidine and 3'-thio-2',3'-dideoxycytidine and the automated synthesis of oligodeoxynucleotides containing a 3'-S-phosphorothiolate linkage

Nucleic Acids Res. 2004 Jan 23;32(2):495-501. doi: 10.1093/nar/gkh189. Print 2004.

Abstract

The synthesis of N4-benzoyl-5'-O-dimethoxytrityl-2',3'-dideoxy-3'-thiocytidine and its phosphorothioamidite is described for the first time, together with a shortened procedure for the preparation of 5'-O-dimethoxytrityl-3'-deoxy-3'-thiothymidine and its corresponding phosphorothioamidite. The first fully automated coupling procedure for the incorporation of a phosphorothioamidite into a synthetic oligodeoxynucleotide has been developed, which conveniently uses routine activators and reagents. Coupling yields using this protocol were in the range of 85-90% and good yields of singularly modified oligonucleotides were obtained. Coupling yields were also equally good when performed on either a 0.2 or 1 micro mol reaction column, thus facilitating large scale syntheses required for mechanistic studies.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Automation / methods
  • Chromatography, High Pressure Liquid
  • Dideoxynucleosides / chemical synthesis
  • Dideoxynucleosides / chemistry*
  • Dideoxynucleotides
  • Oligodeoxyribonucleotides / chemical synthesis*
  • Oligodeoxyribonucleotides / chemistry*
  • Oligodeoxyribonucleotides / isolation & purification
  • Organothiophosphorus Compounds / chemical synthesis
  • Organothiophosphorus Compounds / chemistry
  • Organothiophosphorus Compounds / isolation & purification
  • Phosphates / chemistry*
  • Thionucleosides / chemical synthesis
  • Thionucleosides / chemistry
  • Thymidine / analogs & derivatives*
  • Thymidine / chemical synthesis
  • Thymidine / chemistry*
  • Thymidine / isolation & purification
  • Zalcitabine / analogs & derivatives*
  • Zalcitabine / chemical synthesis
  • Zalcitabine / chemistry*
  • Zalcitabine / isolation & purification

Substances

  • 3'-S-(2-Cyanoethyl-N,N-diisopropylphosphorothioamidite)- 3'-deoxy-5'-O-(4,4'-dimethoxytrityl)thymidine
  • 3'-S-(2-Cyanoethyl-N,N-diisopropylphosphorothioamidite)- N4-benzoyl-3'-deoxy-5'-O-(4,4'-dimethoxytrityl)cytidine
  • 3'-S-phosphorothiolate
  • 3'-thio-2',3'-dideoxycytidine
  • 3'-thio-3'-deoxythymidine
  • Dideoxynucleosides
  • Dideoxynucleotides
  • Oligodeoxyribonucleotides
  • Organothiophosphorus Compounds
  • Phosphates
  • Thionucleosides
  • Zalcitabine
  • Thymidine