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Solid-phase synthesis of oligodeoxynucleotides containing 3'-S-phosphorothiolate linkages
K J Fettes1, I O'Neil, S M Roberts
1Department of Chemistry, University of Liverpool, Crown Street, Liverpool L69 7ZD, United Kingdom.
Nucleosides, Nucleotides & Nucleic Acids
|September 21, 2001
Summary
A new automated method efficiently incorporates 3'-S-phosphorothiolate linkages into DNA using phosphorothioamidite monomers. This breakthrough facilitates large-scale DNA synthesis with high coupling yields.
Area of Science:
- Oligonucleotide Synthesis
- Medicinal Chemistry
Background:
- Phosphorothioate linkages are crucial modifications in DNA therapeutics.
- Existing methods for incorporating these linkages can be complex and difficult to scale.
Purpose of the Study:
- To develop a fully automated procedure for synthesizing DNA with 3 -S-phosphorothiolate linkages.
- To optimize coupling conditions for efficient incorporation of these linkages.
Main Methods:
- Utilized phosphorothioamidite monomers for DNA synthesis.
- Employed activators 5-ethylthiotetrazole and 4,5-dicyanoimidazole.
- Performed automated coupling reactions on varying scales (0.2 and 1 mumole columns).
Main Results:
- Achieved high coupling yields (80-90%) for the phosphorothiolate linkage incorporation.
- Demonstrated consistent yields across different reaction column sizes.
- Successfully automated the entire incorporation process.
Conclusions:
- The developed automated procedure offers an efficient and scalable method for producing DNA containing 3 -S-phosphorothiolate linkages.
- This advancement simplifies the synthesis of modified oligonucleotides for therapeutic applications.
- The high yields and scalability pave the way for broader use in drug development.