Antisense oligonucleotide modulation of human protein kinase C-δ expression

• US 6,235,723 B1
• Filed: 05/18/1999
• Issued: 05/22/2001
• Est. Priority Date: 03/16/1992
• Status: Expired due to Fees

• Current Assignee: ISIS Pharmaceuticals Incorporated
• Original Assignee: ISIS Pharmaceuticals Incorporated
• Inventors: Dean, Nicholas M.
• Primary Examiner(s): Elliott, George C.
• Assistant Examiner(s): ZARA, JANE J
• Application Number:
  US09/313,930
• Time in Patent Office: 735 Days
• Field of Search: 435/6, 435/91.1, 435/91.31, 435/91.51, 435/375, 435/377, 435/455, 435/366, 536/23.1, 536/24.5, 536/25.3, 536/25.6, 514/44
• US Class Current: 514/44.00A
• 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 1/00 Drugs for disorders of the alimentary tract or the digestive system
• CPC Class Codes: A61P 1/16 for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
• CPC Class Codes: A61P 1/18 for pancreatic disorders, e.g. pancreatic enzymes
• CPC Class Codes: A61P 17/00 Drugs for dermatological disorders
• CPC Class Codes: A61P 17/06 Antipsoriatics
• CPC Class Codes: A61P 19/02 for joint disorders, e.g. arthritis, arthrosis
• 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 3/10 for hyperglycaemia, e.g. antidiabetics
• CPC Class Codes: A61P 31/00 Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 35/02 specific for leukemia
• CPC Class Codes: A61P 37/00 Drugs for immunological or allergic disorders
• CPC Class Codes: A61P 37/06 Immunosuppressants, e.g. drugs for graft rejection
• 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 15/1136 against growth factors, growth regulators, cytokines, lymphokines or hormones
• 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 2310/3341 : 5-Methylcytosine
• 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 2310/3525 : MOE, methoxyethoxy
• 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 modulating the expression of PKC-δ and for the treatment and diagnosis of diseases associated with protein kinase C-δ. Methods of treating animals suffering from disease amenable to therapeutic intervention by modulating protein kinase C-δ expression with an oligonucleotide specifically hybridizable with RNA or DNA corresponding to PKC-δ are disclosed. Methods of modulating the expression of TNF-a using the compositions of the present invention are also provided.

Claims

1. An antisense oligonucleotide having 8 to 30 nucleotide units specifically hybridizable with a nucleic acid encoding human PKC-δ

and which is capable of modulating human PKC-δ

expression.

2. The oligonucleotide of claim 1 specifically hybridizable with a translation initiation site or coding region.

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

4. The oligonucleotide of claim 1 wherein at least one of the nucleotides comprises a modification on the 2′

position of the sugar.

5. The oligonucleotide of claim 4 wherein the modification is a 2′

-O-methoxyethyl modification.

6. The oligonucleotide of claim 1 comprising SEQ ID NO:

4, 5, 7, 9, 11, 13, 14, 15 or 16.

7. A pharmaceutical composition comprising the oligonucleotide of claim 1 or claim 6 and a pharmaceutically acceptable carrier or diluent.

8. A pharmaceutical composition comprising the oligonucleotide of claims 1 or 6 and a carrier.

9. A method of inhibiting the expression of human PKC-δ

comprising contacting tissues or cells which express human PKC-δ

in vitro with an effective dose of the oligonucleotide of claim 1 wherein expression of human PKC-δ

is inhibited.

10. The method of claim 9 wherein said expression of human PKC-δ

is abnormal expression.

11. A method of inhibiting hyperproliferation of cells comprising contacting hyperproliferating cells in vitro with an effective dose of the oligonucleotide of claim 1, whereby hyperproliferation of cells is inhibited.

12. A method of modulating the expression of human TNF-α

in cells or tissue comprising contacting said cells or tissue in vitro with the oligonucleotide of claim 1.

13. The method of claim 12 wherein said tissue is adipose tissue.

14. A method of treating an animal having a disease or condition associated with TNF-α

comprising administering to said animal a therapeutically or prophylactically effective amount of an oligonucleotide consisting of the sequence of SEQ ID NO;

19, wherein said disease or condition is diabetes or rheumatoid arthritis, contact hypersensitivity.

15. A method of reducing the blood glucose level in an animal comprising administering to said animal a therapeutically or prophylactically effective amount of an oligonucleotide consisting of the sequence of SEQ ID NO:

19.