Oligonucleotides having phosphorothioate linkages of high chiral purity

• US 5,587,361 A
• Filed: 06/06/1995
• Issued: 12/24/1996
• 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:
  US08/469,851
• Time in Patent Office: 567 Days
• Field of Search: 514/44, 536/22.1, 536/23.1, 536/23.72, 536/24.32, 536/23.7
• 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. A phosphorothioate oligonucleotide complementary to a sequence of targeted RNA or DNA wherein at least 75% of the nucleoside units of said oligonucleotide are joined together by Sp phosphorothioate 3'"'"' to 5'"'"' intersugar linkages.

2. The oligonucleotide of claim 1 wherein said sequence of targeted RNA or DNA to which the oligonucleotide is complementary comprises a portion of a herpes virus genome.

3. The oligonucleotide of claim 1 wherein said sequence of targeted RNA or DNA to which the oligonucleotide is complementary comprises a portion of a papilloma virus genome.

4. The oligonucleotide of claim 1 wherein said sequence of targeted RNA or DNA to which the oligonucleotide is complementary comprises a portion of an HIV genome.

5. The oligonucleotide of claim 1 wherein said oligonucleotide consists of from about 10 to about 50 nucleoside units.

6. The oligonucleotide of claim 1 in a pharmaceutically acceptable carrier.

7. A phosphorothioate oligonucleotide complementary to a sequence of targeted RNA or DNA wherein at least 75% of the nucleoside units of said oligonucleotide are joined together by Rp phosphorothioate 3'"'"' to 5'"'"' intersugar linkages.

8. The oligonucleotide of claim 7 wherein said sequence of targeted RNA or DNA to which the oligonucleotide is complementary comprises a portion of a herpes virus genome.

9. The oligonucleotide of claim 7 wherein said sequence of targeted RNA or DNA to which the oligonucleotide is complementary consists of a portion of a papilloma virus genome.

10. The oligonucleotide of claim 7 wherein said sequence of targeted RNA or DNA to which the oligonucleotide is complementary comprises a portion of an HIV genome.

11. The oligonucleotide of claim 7 wherein said oligonucleotide consists of from about 10 to about 50 nucleoside units.

12. The oligonucleotide of claim 7 in a pharmaceutically acceptable carrier.

13. A phosphorothioate oligonucleotide comprising SEQ ID NO:

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

14. A phosphorothioate oligonucleotide comprising SEQ ID NO:

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

15. A phosphorothioate oligonucleotide comprising SEQ ID NO:

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

16. A phosphorothioate oligonucleotide comprising SEQ ID NO:

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

17. A phosphorothioate oligonucleotide comprising SEQ ID NO:

4, wherein all nucleoside units of said oligonucleotide are joined together by Rp phosphorothioate 3'"'"' to 5'"'"' intersugar linkages.

18. A phosphorothioate oligonucleotide comprising SEQ ID NO:

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