Oligonucleotide modulation of protein kinase C

• US 6,015,892 A
• Filed: 01/11/1996
• Issued: 01/18/2000
• Est. Priority Date: 03/16/1992
• Status: Expired due to Term

• Current Assignee: ISIS Pharmaceuticals Incorporated
• Original Assignee: ISIS Pharmaceuticals Incorporated
• Inventors: Boggs, Russell T., Dean, Nicholas M., Bennett, C. Frank
• Primary Examiner(s): Elliott, George C.
• Assistant Examiner(s): EPPS -SMITH, JANET L
• Application Number:
  US08/578,615
• Time in Patent Office: 1,468 Days
• Field of Search: 514/44, 536/23.1, 536/23.5, 536/24.1, 536/24.5, 536/24.33, 536/24.31, 536/24.3, 435/6
• US Class Current: 536/24.5
• 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: 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: A61P 29/00 Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• 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: C07K 14/70564 Selectins, e.g. CD62
• CPC Class Codes: C12N 15/1137 against enzymes viral enzymes C12N15/1131; receptors C12N15/1138
• CPC Class Codes: C12N 2310/315 Phosphorothioates
• CPC Class Codes: C12N 2310/321 2'-O-R Modification
• CPC Class Codes: C12N 2310/322 2'-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: C12N 2310/3527 Other alkyl chain
• CPC Class Codes: C12N 2310/3533 Halogen
• CPC Class Codes: C12N 9/12 transferring phosphorus containing groups, e.g. kinases (2.7)
• 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 1/6883 for diseases caused by alterations of genetic material
• CPC Class Codes: C12Q 2600/158 Expression markers
• CPC Class Codes: C12Y 207/11013 : Protein kinase C (2.7.11.13)

Abstract

Compositions and methods are provided for the treatment and diagnosis of diseases associated with protein kinase C. Oligonucleotides are provided which are specifically hybridizable with a PKC gene or mRNA. Oligonucleotides specifically hybridizable with a particular PKC isozyme, set of isozymes or mRNA transcript are provided. Methods of treating conditions amenable to therapeutic intervention by modulating protein kinase C expression with an oligonucleotide specifically hybridizable with a PKC gene or mRNA are disclosed. Compositions and methods are provided for the treatment, detection and diagnosis of diseases associated with protein kinase C alpha and specific transcripts thereof. New nucleic acid sequences are provided which encode 3'"'"' untranslated regions of human protein kinase C alpha polynucleotide probes for PKC alpha are also disclosed.

Claims

1. An oligonucleotide up to 50 nucleotide units in length comprising a nucleotide sequence selected from the group consisting of SEQ ID NO:

1, 2, 4, 5, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 18, 19, 20, 52 and 53.

2. The oligonucleotide of claim 1 wherein at least one of the intersugar linkages between nucleotide units of the oligonucleotide is a phosphorothioate.

3. The oligonucleotide of claim 1 wherein at least one of the nucleotide units comprises a modification on the 2'"'"' position of the sugar.

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

5. An isolated nucleic acid molecule comprising the nucleotide sequence of SEQ ID NO:

105.

6. The nucleic acid molecule of claim 5 wherein said nucleic acid molecule is double-stranded.

8. A polynucleotide probe up to 50 nucleotide units in length comprising a nucleotide sequence complementary to a portion of the nucleic acid molecule of claim 5 or claim 7.

7. An isolated nucleic acid molecule comprising a sequence complementary to the nucleotide sequence of SEQ ID NO:

105.

9. An antisense oligonucleotide 5 to 50 nucleotides in length comprising a nucleotide sequence complementary to a portion of the sequence set forth in SEQ ID NO:

105.

10. An antisense oligonuclcotide 5 to 50 nucleotides in length comprising a nucleotide sequence which is complementary to the long mRNA transcript of human protein kinase C-α

and which is not complementary to the short mRNA transcript of human protein kinase C-α

.

11. The anti sense oligonucleotide of claim 10 comprising a nucleotide sequence complementary to a portion of the sequence set forth in SEQ ID NO:

106.

12. The oligonucleotide of claim 11 comprising a sequence as set forth in SEQ ID NO:

117.

13. A polynucleotide probe comprising a nuclcotide sequence complementary to the long mRNA transcript of human protein kinase C-α

.

14. The polynucleotide probe of claim 13 comprising a sequence as set forth in SEQ ID NO:

117.

15. A method of inhibiting the expression of human protein kinase C-α

in vitro comprising contacting human cells with a therapeutically effective amount of an antisense oligonucleotide 5 to 50 nucleotides in length, said antisense oligonucleotide comprising a nucleotide sequence complementary to a portion of the sequence set forth in SEQ ID NO;

105.

16. A method of inhibiting the expression of human protein kinase C-α

in vitro comprising contacting human cells with a therapeutically effective amount of an antisense oligonucleotide 5 to 50 nucleotides in length, said antisense oligonucleotide comprising a nucleotide sequence complementary to a portion of the sequence set forth in SEQ ID NO;

106.

17. An oligonucleotide up to 50 nucleotide units in length comprising a nucleotide sequence selected from the group consisting of SEQ ID NO:

3, 6 and 17.

18. The oligonucleotide of claim 17 wherein at least one of the nucleotide units comprises a modification on the 2'"'"' position of the sugar.

19. The oligonucleotide of claim 17 wherein at least one of the intersugar linkages between nucleotide units of the oligonucleotide is a phosphorothioate.

20. The oligonucleotide of claim 17 which is a chimeric oligonucleotide.