Oligonucleotides having a conserved G4 core sequence

• US 5,952,490 A
• Filed: 06/12/1995
• Issued: 09/14/1999
• Est. Priority Date: 09/29/1992
• Status: Expired due to Term

• Current Assignee: ISIS Pharmaceuticals Incorporated
• Original Assignee: ISIS Pharmaceuticals Incorporated
• Inventors: Brown-Driver, Vickie L., Chiang, Ming-Yi, Imbach, Jean Louis, Bennett, C. Frank, Ecker, David J., Vickers, Timothy A., Hanecak, Ronnie C., Anderson, Kevin P., Wyatt, Jacqueline R.
• Primary Examiner(s): Guzo, David
• Application Number:
  US08/403,888
• Time in Patent Office: 1,555 Days
• Field of Search: 435/69.1, 435/172.1, 435/172.3, 435/320.1, 514/44, 536/23.1, 536/24.5, 536/25.5
• US Class Current: 536/24.5
• CPC Class Codes: A61K 31/70 Carbohydrates; Sugars; Derivatives thereof sorbitol A61K31/047
• CPC Class Codes: A61K 38/00 Medicinal preparations containing peptides peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00
• CPC Class Codes: A61P 31/12 Antivirals
• CPC Class Codes: A61P 31/16 for influenza or rhinoviruses
• CPC Class Codes: A61P 31/18 for HIV
• CPC Class Codes: A61P 31/22 for herpes viruses
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• 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: C12N 15/1133 against herpetoviridae, e.g. HSV
• CPC Class Codes: C12N 15/1137 against enzymes viral enzymes C12N15/1131; receptors C12N15/1138
• CPC Class Codes: C12N 15/115 Aptamers, i.e. nucleic acids binding a target molecule specifically and with high affinity without hybridising therewith ; Nucleic acids binding to non-nucleic acids, e.g. aptamers
• CPC Class Codes: C12N 15/117 Nucleic acids having immunomodulatory properties, e.g. containing CpG-motifs
• CPC Class Codes: C12N 2310/151 more than 3 strands, e.g. tetrads, H-DNA
• CPC Class Codes: C12N 2310/18 acting by a non-sequence specific mechanism other than C12N2310/16 or C12N2310/17
• CPC Class Codes: C12N 2310/315 Phosphorothioates
• CPC Class Codes: C12N 2310/321 2'-O-R Modification
• CPC Class Codes: C12N 2310/341 Gapmers, i.e. of the type ===---===
• CPC Class Codes: C12N 2310/346 having a combination of backbone and sugar modifications
• CPC Class Codes: C12N 2310/3521 Methyl
• CPC Class Codes: C12Q 1/701 Specific hybridization probes
• CPC Class Codes: C12Y 207/07049 : RNA-directed DNA polymerase (2.7.7.49), i.e. telomerase or reverse-transcriptase
• CPC Class Codes: C12Y 301/01004 : Phospholipase A2 (3.1.1.4)

Abstract

Modified oligonucleotides having a conserved G₄ sequence and a sufficient number of flanking nucleotides to significantly inhibit the activity of a virus such as HSV-1 or phospholipase A₂ or to modulate the telomere length of a chromosome are provided. G₄ quartet oligonucleotide structures are also provided. Methods of prophylaxis, diagnosis and therapeutics for viral-associated diseases and diseases associated with elevated levels of phospholipase A₂ are also provided. Methods of modulating telomere length of a chromosome are also provided; modulation of telomere length is believed to plat a role in the aging process of a cell and in control of malignant cell growth.

Claims

1. A chemically modified oligonucleotide having no more than about 27 nucleic acid base units, said oligonucleotide comprising at least one GGGG sequence or at least two GGG sequences and a sufficient number of flanking nucleotides to significantly inhibit the activity of a virus or phospholipase A₂.

2. The oligonucleotide of claim 1 wherein significant inhibition of viral or enzyme activity is at least 50% inhibition.

3. The oligonucleotide of claim 1 wherein the virus is human immunodeficiency virus, herpes simplex virus, human cytomegalovirus or influenza virus.

4. The oligonucleotide of claim 1 which has at least one phosphorothioate intersugar (backbone) linkage.

5. The oligonucleotide of claim 1 wherein each of the nucleotides is in the alpha (α

) anomeric configuration.

6. The oligonucleotide of claim 1 which is a chimeric oligonucleotide.

7. The oligonucleotide of claim 3 wherein the virus is herpes simplex virus.

8. The oligonucleotide of claim 7 wherein the oligonucleotide is selected from the group consisting of:

SEQ ID NO;

21, SEQ ID NO;

1, SEQ ID NO;

8, SEQ ID NO;

12, SEQ ID NO;

16, SEQ ID NO;

22, SEQ ID NO;

28, SEQ ID NO;

30, SEQ ID NO;

31, SEQ ID NO;

33, SEQ ID NO;

35, SEQ ID NO;

36, SEQ ID NO;

37, SEQ ID NO;

38, SEQ ID NO;

39, SEQ ID NO;

42, SEQ ID NO;

47, SEQ ID NO;

48 and SEQ ID NO;

50.

9. The oligonucleotide of claim 7 having a sequence shown in Table 8.

10. The oligonucleotide of claim 9 having a sequence selected from the group consisting of SEQ ID NO:

124, SEQ ID NO;

126, SEQ ID NO;

127, SEQ ID NO;

128, SEQ ID NO;

129, SEQ ID NO;

130 and SEQ ID NO;

133.

11. A chemically modified oligonucleotide having the sequence (N_X G₄ N_Y)_Q wherein X and Y are independently 1 to 8 and Q is 1 to 4, wherein said oligonucleotide significantly inhibits the activity of a virus or phospholipase A₂.

12. The oligonucleotide of claim 11 having the sequence NNGGGGNN.

13. The oligonucleotide of claim 12 which has at least one phosphorothioate intersugar linkage and which has the sequence GNGGGGTN.

14. A chemically modified oligonucleotide having the sequence (G₄ N_X G₄)_Q wherein X is 1 to 8 and Q is 1 to 3, wherein said oligonucleotide significantly inhibits the activity of a virus or phospholipase A₂.

15. A chemically modified oligonucleotide having the sequence (N_X G_3-4)_Q N_X wherein X is 1 to 8 and Q is 1 to 6, wherein said oligonucleotide significantly inhibits the activity of a virus or phospholipase A₂.

16. A phosphorothioate oligonucleotide having SEQ ID NO:

21.

17. A phosphorothioate oligonucleotide having the sequence TTGGGGTT.

18. The oligonucleotide of claim 17 wherein each of the nucleotides of the oligonucleotide is in the alpha (α

) anomeric configuration.

19. A compound comprising a G-quartet structure of phosphorothioate oligonucleotides each oligonucleotide having the sequence TxG4Ty, wherein x is 2 and y is 2, or x is 0 and y is 2, or x is 3 and v is 3.

20. The compound of claim 19 wherein the nucleotides of at least one of the oligonucleotides of the G-quartet structure are in the alpha (α

) anomeric configuration.

21. The compound of claim 19 wherein x is 2 and y is 2.

22. The compound of claim 19 wherein x is 0 and y is 2.

23. The compound of claim 19 wherein x is 3 and y is 3.

24. A chemically modified oligonucleotide having no more than about 27 nucleic acid base units, said oligonucleotide comprising at least two GGG sequences and a sufficient number of flanking nucleotides to modulate the telomere length of a chromosome.

25. The oligonucleotide of claim 24 which has at least one phosphorothioate intersugar linkage.

26. The oligonucleotide of claim 24 wherein each of the nucleotides is in the alpha (α

) anomeric configuration.

27. The oligonucleotide of claim 24 which is a chimeric oligonucleotide.