Skip to main content
NCBI home page
Nucleic Acids Research logoLink to Nucleic Acids Research
. 1987 Dec 10;15(23):9921–9932. doi: 10.1093/nar/15.23.9921

An approach to the stereoselective synthesis of Sp-dinucleoside phosphorothioates using phosphotriester chemistry.

R Cosstick 1, D M Williams 1
PMCID: PMC306540  PMID: 3480505

Abstract

An approach to the stereoselective synthesis of Sp- dinucleoside phosphorothioates has been investigated which utilizes phosphotriester chemistry. The stereoselectivity of internucleotide bond formation between N4-benzoyl-5'-O-(4,4'-dimethoxytrityl)-2'-deoxycytidine-3'-O-(S2-cyano-e thyl) phosphorothioate (3) and 3'-O-acetylthymidine has been studied using either mesitylenesulphonyl-5-(pyridin-2-yl)tetrazole (MSPy) or 1-mesitylenesulphonyl-3-nitro-1,2,4-triazole (MSNT) as the activating agent. The removal of the cyanoethyl group from the protected dinucleoside phosphorothioate has been studied, and conditions are reported which provide rapid deprotection without concomittant desulphurisation.

Full text

PDF
9921

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Belagaje R., Brush C. K. Polymer supported synthesis of oligonucleotides by a phosphotriester method. Nucleic Acids Res. 1982 Oct 25;10(20):6295–6603. doi: 10.1093/nar/10.20.6295. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Burgers P. M., Eckstein F. Diastereomers of 5'-O-adenosyl 3'-O-uridyl phosphorothioate: chemical synthesis and enzymatic properties. Biochemistry. 1979 Feb 20;18(4):592–596. doi: 10.1021/bi00571a007. [DOI] [PubMed] [Google Scholar]
  3. Connolly B. A., Eckstein F., Pingoud A. The stereochemical course of the restriction endonuclease EcoRI-catalyzed reaction. J Biol Chem. 1984 Sep 10;259(17):10760–10763. [PubMed] [Google Scholar]
  4. Connolly B. A., Potter B. V., Eckstein F., Pingoud A., Grotjahn L. Synthesis and characterization of an octanucleotide containing the EcoRI recognition sequence with a phosphorothioate group at the cleavage site. Biochemistry. 1984 Jul 17;23(15):3443–3453. doi: 10.1021/bi00310a010. [DOI] [PubMed] [Google Scholar]
  5. Cosstick R., Eckstein F. Synthesis of d(GC) and d(CG) octamers containing alternating phosphorothioate linkages: effect of the phosphorothioate group on the B-Z transition. Biochemistry. 1985 Jul 2;24(14):3630–3638. doi: 10.1021/bi00335a035. [DOI] [PubMed] [Google Scholar]
  6. Cosstick R., McLaughlin L. W., Eckstein F. Fluorescent labelling of tRNA and oligodeoxynucleotides using T4 RNA ligase. Nucleic Acids Res. 1984 Feb 24;12(4):1791–1810. doi: 10.1093/nar/12.4.1791. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Cruse W. B., Salisbury S. A., Brown T., Cosstick R., Eckstein F., Kennard O. Chiral phosphorothioate analogues of B-DNA. The crystal structure of Rp-d[Gp(S)CpGp(S)CpGp(S)C]. J Mol Biol. 1986 Dec 20;192(4):891–905. doi: 10.1016/0022-2836(86)90035-5. [DOI] [PubMed] [Google Scholar]
  8. Duckworth M. L., Gait M. J., Goelet P., Hong G. F., Singh M., Titmas R. C. Rapid synthesis of oligodeoxyribonucleotides VI. Efficient, mechanised synthesis of heptadecadeoxyribonucleotides by an improved solid phase phosphotriester route. Nucleic Acids Res. 1981 Apr 10;9(7):1691–1706. doi: 10.1093/nar/9.7.1691. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Eckstein F. Nucleoside phosphorothioates. Annu Rev Biochem. 1985;54:367–402. doi: 10.1146/annurev.bi.54.070185.002055. [DOI] [PubMed] [Google Scholar]
  10. Gait M. J., Popov S. G., Singh M., Titmas R. C. Rapid synthesis of oligodeoxyribonucleotides. V. Further studies in solid phase synthesis of oligodeoxyribonucleotides through phosphotriester intermediates. Nucleic Acids Symp Ser. 1980;(7):243–257. [PubMed] [Google Scholar]
  11. Gough G. R., Singleton C. K., Weith H. L., Gilham P. T. Protected deoxyribonucleoside-3' aryl phosphodiesters as key intermediates in polynucleotide synthesis. Construction of an icosanucleotide analogous to the sequence at the ends of Rous sarcoma virus 35S RNA. Nucleic Acids Res. 1979 Apr;6(4):1557–1570. doi: 10.1093/nar/6.4.1557. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Hsiung H., Inouye S., West J., Sturm B., Inouye M. Further improvements on the phosphotriester synthesis of deoxyribooligonucleotides and the oligonucleotide directed site-specific mutagenesis of E. coli lipoprotein gene. Nucleic Acids Res. 1983 May 25;11(10):3227–3239. doi: 10.1093/nar/11.10.3227. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. LaPlanche L. A., James T. L., Powell C., Wilson W. D., Uznanski B., Stec W. J., Summers M. F., Zon G. Phosphorothioate-modified oligodeoxyribonucleotides. III. NMR and UV spectroscopic studies of the Rp-Rp, Sp-Sp, and Rp-Sp duplexes, [d(GGSAATTCC)]2, derived from diastereomeric O-ethyl phosphorothioates. Nucleic Acids Res. 1986 Nov 25;14(22):9081–9093. doi: 10.1093/nar/14.22.9081. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Marugg J. E., van den Bergh C., Tromp M., van der Marel G. A., van Zoest W. J., van Boom J. H. Synthesis of phosphorothioate-containing DNA fragments by a modified hydroxybenzotriazole phosphotriester approach. Nucleic Acids Res. 1984 Dec 11;12(23):9095–9110. doi: 10.1093/nar/12.23.9095. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Ohtsuka E., Tozuka Z., Iwai S., Ikehara M. A new condensing reagent, 1-(2,4,6-triisopropylbenzenesulfonyl)-5-(pyridin-2-yl)tetrazolide and its use in the synthesis of lambda cro binding heptadecanucleotide on a polymer support. Nucleic Acids Res. 1982 Oct 25;10(20):6235–6241. doi: 10.1093/nar/10.20.6235. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Potter B. V., Connolly B. A., Eckstein F. Synthesis and configurational analysis of a dinucleoside phosphate isotopically chiral at phosphorus. Stereochemical course of Penicillium citrum nuclease P1 reaction. Biochemistry. 1983 Mar 15;22(6):1369–1377. doi: 10.1021/bi00275a008. [DOI] [PubMed] [Google Scholar]
  17. Romaniuk P. J., Eckstein F. A study of the mechanism of T4 DNA polymerase with diastereomeric phosphorothioate analogues of deoxyadenosine triphosphate. J Biol Chem. 1982 Jul 10;257(13):7684–7688. [PubMed] [Google Scholar]
  18. Sinha N. D., Biernat J., McManus J., Köster H. Polymer support oligonucleotide synthesis XVIII: use of beta-cyanoethyl-N,N-dialkylamino-/N-morpholino phosphoramidite of deoxynucleosides for the synthesis of DNA fragments simplifying deprotection and isolation of the final product. Nucleic Acids Res. 1984 Jun 11;12(11):4539–4557. doi: 10.1093/nar/12.11.4539. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Sood A. K., Narang S. A. A rapid and convenient synthesis of poly-thymidylic acid by the modified triester approach. Nucleic Acids Res. 1977 Aug;4(8):2757–2765. doi: 10.1093/nar/4.8.2757. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Nucleic Acids Research are provided here courtesy of Oxford University Press

RESOURCES