Oligonucleotides for modulating hepatitis C virus having phosphorothioate linkages of high chiral purity

• US 5,576,302 A
• Filed: 06/06/1995
• Issued: 11/19/1996
• Est. Priority Date: 10/15/1991
• Status: Expired due to Term

• Current Assignee: ISIS Pharmaceuticals Incorporated (Ionis Pharmaceuticals, Inc.)
• Original Assignee: ISIS Pharmaceuticals Incorporated (Ionis Pharmaceuticals, Inc.)
• Inventors: Cook, Phillip D., Hoke, Glenn
• Primary Examiner(s): Wilson, James O.
• Application Number:
  US08/468,447
• Time in Patent Office: 532 Days
• Field of Search: 514/42, 514/43, 514/44, 514/45, 514/46, 536/24.5, 536/25.33, 536/25.34
• US Class Current: 514/44.00A
• CPC Class Codes: A61K 31/70 Carbohydrates; Sugars; Derivatives thereof sorbitol A61K31/047
• CPC Class Codes: A61K 48/00 Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy
• CPC Class Codes: C07H 19/04 Heterocyclic radicals containing only nitrogen atoms as ring hetero atom
• CPC Class Codes: C07H 19/06 Pyrimidine radicals
• CPC Class Codes: C07H 19/10 with the saccharide radical esterified by phosphoric or polyphosphoric acids
• CPC Class Codes: C07H 19/16 Purine radicals
• CPC Class Codes: C07H 19/20 with the saccharide radical esterified by phosphoric or polyphosphoric acids
• CPC Class Codes: C07H 21/00 Compounds containing two or more mononucleotide units having separate phosphate or polyphosphate groups linked by saccharide radicals of nucleoside groups, e.g. nucleic acids
• CPC Class Codes: C07H 23/00 Compounds containing boron, silicon, or a metal, e.g. chelates, vitamin B12 esters with inorganic acids C07H11/00; metal salts, see parent compounds
• CPC Class Codes: C07J 43/003 not condensed
• CPC Class Codes: C12N 15/10 Processes for the isolation, preparation or purification of DNA or RNA chemical preparation of DNA or RNA C07H21/00; preparation of non-structural polynucleotides from microorganisms or with enzymes C12P19/34
• CPC Class Codes: C12N 15/113 Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing when used in plants C12N15/8218
• CPC Class Codes: C12N 15/1131 against viruses
• CPC Class Codes: C12N 15/1132 against retroviridae, e.g. HIV
• CPC Class Codes: C12N 15/1133 against herpetoviridae, e.g. HSV
• CPC Class Codes: C12N 15/1135 against oncogenes or tumor suppressor genes
• CPC Class Codes: C12N 15/1137 against enzymes viral enzymes C12N15/1131; receptors C12N15/1138
• CPC Class Codes: C12N 2310/3125 Methylphosphonates
• CPC Class Codes: C12N 2310/315 Phosphorothioates
• CPC Class Codes: C12N 2310/32 of the sugar
• CPC Class Codes: C12N 2310/321 : 2'-O-R Modification
• CPC Class Codes: C12N 2310/322 : 2'-R Modification
• CPC Class Codes: C12N 2310/33 : of the base
• CPC Class Codes: C12N 2310/333 : Modified A
• CPC Class Codes: C12N 2310/336 : Modified G
• CPC Class Codes: C12N 2310/3521 : Methyl
• CPC Class Codes: C12N 2310/3523 : Allyl
• CPC Class Codes: C12N 2310/3527 : Other alkyl chain
• CPC Class Codes: C12N 2310/3531 : Hydrogen
• CPC Class Codes: C12N 2310/3533 : Halogen
• CPC Class Codes: C12N 9/0069 : acting on single donors with incorporation of molecular oxygen, i.e. oxygenases (1.13)
• CPC Class Codes: C12P 19/34 : Polynucleotides, e.g. nucleic acids, oligoribonucleotides
• CPC Class Codes: C12Q 1/68 : involving nucleic acids
• CPC Class Codes: C12Q 1/6813 : Hybridisation assays
• CPC Class Codes: C12Q 1/6876 : Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
• CPC Class Codes: C12Q 1/701 : Specific hybridization probes
• CPC Class Codes: C12Y 113/11012 : Linoleate 13S-lipoxygenase (1.13.11.12)

Abstract

Sequence-specific phosphorothioate oligonucleotides comprising nucleoside units which are joined together by either substantially all Sp or substantially all Rp phosphorothioate intersugar linkages are provided. Such sequence-specific phosphorothioate oligonucleotides having substantially chirally pure intersugar linkages are prepared by enzymatic or chemical synthesis.

Claims

1. An oligonucleotide represented by SEQ ID NO:

1 wherein at least 75% of the nucleoside units are joined together by Sp phosphorothioate 3'"'"' to 5'"'"' linkages.

3. The oligonucleotide of claim 1 wherein all of the nucleoside units are joined together by Sp phosphorothioate 3'"'"' to 5'"'"' linkages.

21. A composition containing an oligonucleotide of claim 1 and an acceptable carrier.

2. An oligonucleotide represented by SEQ ID NO:

1 wherein at least 75% of the nucleoside units are joined together by Rp phosphorothioate 3'"'"' to 5'"'"' linkages.

4. The oligonucleotide of claim 2 wherein all of the nucleoside units are joined together by Rp phosphorothioate 3'"'"' to 5'"'"' linkages.

22. A composition containing an oligonucleotide of claim 2 and an acceptable carrier.

5. An oligonucleotide represented by SEQ ID NO:

2 wherein at least 75% of the nucleoside units are joined together by Sp phosphorothioate 3'"'"' to 5'"'"' linkages.

7. The oligonucleotide of claim 5 wherein all of the nucleoside units are joined together by Sp phosphorothioate 3'"'"' to 5'"'"' linkages.

23. A composition containing an oligonucleotide of claim 5 and an acceptable carrier.

6. An oligonucleotide represented by SEQ ID NO:

2 wherein at least 75% of the nucleoside units are joined together by Rp phosphorothioate 3'"'"' to 5'"'"' linkages.

8. The oligonucleotide of claim 6 wherein all of the nucleoside units are joined together by Rp phosphorothioate 3'"'"' to 5'"'"' linkages.

24. A composition containing an oligonucleotide of claim 6 and an acceptable carrier.

9. An oligonucleotide represented by SEQ ID NO:

3 wherein at least 75% of the nucleoside units are joined together by Sp phosphorothioate 3'"'"' to 5'"'"' linkages.

11. The oligonucleotide of claim 9 wherein all of the nucleoside units are joined together by Sp phosphorothioate 3'"'"' to 5'"'"' linkages.

25. A composition containing an oligonucleotide of claim 9 and an acceptable carrier.

10. An oligonucleotide represented by SEQ ID NO:

3 wherein at least 75% of the nucleoside units are joined together by Rp phosphorothioate 3'"'"' to 5'"'"' linkages.

12. The oligonucleotide of claim 10 wherein all of the nucleoside units are joined together by Rp phosphorothioate 3'"'"' to 5'"'"' linkages.

26. A composition containing an oligonucleotide of claim 10 and an acceptable carrier.

13. An oligonucleotide represented by SEQ ID NO:

4 wherein at least 75% of the nucleoside units are joined together by Sp phosphorothioate 3'"'"' to 5'"'"' linkages.

15. The oligonucleotide of claim 13 wherein all of the nucleoside units are joined together by Sp phosphorothioate 3'"'"' to 5'"'"' linkages.

27. A composition containing an oligonucleotide of claim 13 and an acceptable carrier.

14. An oligonucleotide represented by SEQ ID NO:

4 wherein at least 75% of the nucleoside units are joined together by Rp phosphorothioate 3'"'"' to 5'"'"' linkages.

16. The oligonucleotide of claim 14 wherein all of the nucleoside units are joined together by Rp phosphorothioate 3'"'"' to 5'"'"' linkages.

28. A composition containing an oligonucleotide of claim 14 and an acceptable carrier.

17. An oligonucleotide represented by SEQ ID NO:

5 wherein at least 75% of the nucleoside units are joined together by Sp phosphorothioate 3'"'"' to 5'"'"' linkages.

19. The oligonucleotide of claim 17 wherein all of the nucleoside units are joined together by Sp phosphorothioate 3'"'"' to 5'"'"' linkages.

29. A composition containing an oligonucleotide of claim 17 and an acceptable carrier.

18. An oligonucleotide represented by SEQ ID NO:

5 wherein at least 75% of the nucleoside units are joined together by Rp phosphorothioate 3'"'"' to 5'"'"' linkages.

20. The oligonucleotide of claim 18 wherein all of the nucleoside units are joined together by Rp phosphorothioate 3'"'"' to 5'"'"' linkages.

30. A composition containing an oligonucleotide of claim 18 and an acceptable carrier.