Enhanced antisense modulation of ICAM-1

• US 6,111,094 A
• Filed: 04/17/1998
• Issued: 08/29/2000
• Est. Priority Date: 08/14/1990
• Status: Expired due to Fees

• Current Assignee: ISIS Pharmaceuticals Incorporated (Ionis Pharmaceuticals, Inc.)
• Original Assignee: ISIS Pharmaceuticals Incorporated (Ionis Pharmaceuticals, Inc.)
• Inventors: Bennett, C. Frank, Condon, Thomas P., Flournoy, Shin Cheng
• Primary Examiner(s): Elliot, George C.
• Assistant Examiner(s): EPPS -SMITH, JANET L
• Application Number:
  US09/062,416
• Time in Patent Office: 865 Days
• Field of Search: 435/6, 435/91.5, 435/375, 435/325, 435/366, 536/23.1, 536/24.3, 536/24.31, 536/24.33, 536/24.5, 514/44
• 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 31/12 Antivirals
• CPC Class Codes: A61P 31/18 for HIV
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: A61P 9/10 for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
• CPC Class Codes: C12N 15/1138 against receptors or cell surface proteins
• CPC Class Codes: C12N 2310/321 2'-O-R Modification
• CPC Class Codes: C12N 2310/322 2'-R Modification
• CPC Class Codes: C12N 2310/334 Modified C
• 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 2310/3525 MOE, methoxyethoxy

Abstract

The present invention provides compositions and methods for detecting and modulating levels of intercellular adhesion molecule-1 (ICAM-1) proteins, including human ICAM-1.

Claims

1. An antisense oligonucleotide comprising from about 8 to about 30 nucleotides connected by covalent linkages, wherein said antisense oligonucleotide comprises a nucleobase sequence specifically hybridizable with a nucleic acid encoding a human intercellular adhesion molecule-1, wherein said antisense compound modulates the expression of said human intercellular adhesion molecule-1 and wherein said oligonucleotide comprises at least one 2'"'"'-modification.

2. The antisense oligonucleotide of claim 1 wherein said 2'"'"'-modification is a 2'"'"'-methoxyethoxy modification.

3. The antisense oligonucleotide of claim 2 wherein said 2'"'"'-methoxyethoxy modification is present on the most 3'"'"' residue of said oligonucleotide.

4. The antisense oligonucleotide of claim 3, wherein every cytosine nucleobase of said oligonucleotide is substituted by a 5-methyl cytosine nucleobase.

5. The antisense oligonucleotide of claim 1, further comprising at least one 5-methyl cytosine nucleobase in place of a cytosine nucleobase.

6. The antisense oligonucleotide of claim 1 wherein said cellular adhesion molecule-1 is human cellular adhesion molecule-1.

7. The antisense oligonucleotide of claim 6 wherein said nucleobase sequence is SEQ ID NO:

2, 5, 6, 7 or 8.

8. A pharmaceutical composition comprising the antisense oligonucleotide of claim 6 and a pharmaceutically acceptable carrier.

9. A method of modulating the synthesis of intercellular adhesion molecule-1 in a cell or tissue comprising contacting the cell or tissue in-vitro with the antisense oligonucleotide of claim 1.

10. An antisense oligonucleotide that modulates the expression of human intercellular adhesion molecule-1, wherein said oligonucleotide has the structure:

GsCsCsCsAsAsGsCsTsGsGsCsAsTsCsGsTsCsA,

wherein;

"s" is a phosphorothioate linkage;

"A", "C", "T" and "G" are 2'"'"'-deoxy-adenoine, cytidine, thymidine and guanosine nucleoside residues, respectively;

"A" is an adenosine nucleoside residue comprising a 2'"'"' modification;

"T" is a thymidine nucleoside residue comprising a 2'"'"' modification;

"G" is a guanosine nucleoside residue comprising a 2'"'"' modification; and

"C" is a 5-methyl-cytidine nucleoside residue comprising a 2'"'"' modification.

11. The antisense oligonucleotide of claim 10, wherein said 2'"'"'-modification is 2'"'"'-methoxyethoxy and said oligonucleotide is ISIS 13280.

12. The antisense oligonucleotide of claim 10, wherein said 2'"'"'-modification is 2'"'"'-fluoro and said oligonucleotide is ISIS 12604.

13. A pharmaceutical composition comprising the antisense oligonucleotide of claim 10 and a pharmaceutically acceptable carrier.

14. An antisense oligonucleotide that modulates the expression of human intercellular adhesion molecule-1, wherein said oligonucleotide is ISIS 14725 having the structure:

GsCsCsCsAsAsGsCsTsGsGsCsAsTsCsCsGsTsCsA,

wherein;

"s" is a phosphorothioate linkage;

"A", "C", "T" and "G" are 2'"'"'-deoxy-adenosine, cytidine, thymidine and guanosine nucleoside residues, respectively;

"A" is a 2'"'"'-methoxyethoxy adenosine nucleoside residue;

"T" is a 2'"'"'-methoxyethoxy thymidine nucleoside residue;

"G" is a 2'"'"'-methoxyethoxy guanosine nucleoside residue; and

"C" is a 2'"'"'-methoxyethoxy 5-methyl-cytidine nucleoside residue.

15. A pharmaceutical composition comprising the antisense oligonucleotide of claim 14 and a pharmaceutically acceptable carrier.

16. An antisense oligonucleotide that modulates the expression of human intercellular adhesion molecule-1, wherein said oligonucleotide is ISIS 15839 having the structure:

GsCsCsCsAsAsGsCsTsGsGsCsAsTsCsGsTsCsA,

wherein;

"s" is a phosphorothioate linkage;

"A", "T" and "G" are 2'"'"'-deoxy-adenosine, thymidine and guanosine nucleoside residues, respectively;

"C" is a 2'"'"'-deoxy-5-methyl-cytidine nucleoside residue;

"A"is a 2'"'"'-methoxyethoxy adenosine nucleoside residue;

"T" is a 2'"'"'-methoxyethoxy thymidine nucleoside residue;

"G" is a 2'"'"'-methoxyethoxy guanosine nucleoside residue; and

"C" is a 2'"'"'-methoxyethoxy 5-methyl-cytidine nucleoside residue.

17. A pharmaceutical composition comprising the antisense oligonucleotide of claim 16 and a pharmaceutically acceptable carrier.

18. An antisense oligonucleotide that modulates the expression of human intercellular adhesion molecule-1, wherein said oligonucleotide is ISIS 16824 having the structure:

CsCsAsAsGsCsTsGsGsCsAsTsCsCsGsTsCsA,

wherein;

"s" is a phosphorothioate linkage;

"A", "C", "T" and "G" are 2'"'"'-deoxy-adenosine, cytidine, thymidine and guanosine nucleoside residues, respectively;

"A" is a 2'"'"'-methoxyethoxy adenosine nucleoside residue;

"T" is a 2'"'"'-methoxyethoxy thymidine nucleoside residue;

"G" is a 2'"'"'-methoxyethoxy guanosine nucleoside residue; and

"C" is a 2'"'"'-methoxyethoxy 5-methyl-cytidine nucleoside residue.

19. A pharmaceutical composition comprising the antisense oligonucleotide of claim 18 and a pharmaceutically acceptable carrier.

20. An antisense oligonucleotide that modulates the expression of human intercellular adhesion molecule-1, wherein said oligonucleotide is ISIS 16823 having the structure:

AsAsGsCsTsGsGsCsAsTsCsCsGsTsCsA,

wherein;

"s" is a phosphorothioate linkage;

"A", "C", "T" and "G" are 2'"'"'-deoxy-adenosine, cytidine, thymidine and guanosine nucleoside residues, respectively;

"A" is a 2'"'"'-methoxyethoxy adenosine nucleoside residue;

"T" is a 2'"'"'-methoxyethoxy thymidine nucleoside residue;

"G" is a 2'"'"'-methoxyethoxy guanosine nucleoside residue; and

"C" is a 2'"'"'-methoxyethoxy 5-methyl-cytidine nucleoside residue.

21. A pharmaceutical composition comprising the antisense oligonucleotide of claim 20 and a pharmaceutically acceptable carrier.

22. An antisense oligonucleotide that modulates the expression of human intercellular adhesion molecule-1, wherein said oligonucleotide is ISIS 16158 having the structure:

GsTsGsCsCsCsAsAsGsCsTsGsGsCsAsTsCsCsGsT,

wherein;

"s" is a phosphorothioate linkage;

"A", "C", "T" and "G" are 2'"'"'-deoxy-adenosine, cytidine, thymidine and guanosine nucleoside residues, respectively;

"A" is a 2'"'"'-methoxyethoxy adenosine nucleoside residue;

"T" is a 2'"'"'-methoxyethoxy thymidine nucleoside residue;

"G" is a 2'"'"'-methoxyethoxy guanosine nucleoside residue; and

"C" is a 2'"'"'-methoxyethoxy 5-methyl-cytidine nucleoside residue.

23. A pharmaceutical composition comprising the antisense oligonucleotide of claim 22 and a pharmaceutically acceptable carrier.

24. An antisense oligonucleotide that modulates the expression of human intercellular adhesion molecule-1, wherein said oligonucleotide is ISIS 16159 having the structure:

AsGsCsAsGsTsGsTsGsCsCsCsAsAsGsCsTsGsGsCsAsT,

wherein;

"s" is a phosphorothioate linkage;

"A", "C", "T" and "G" are 2'"'"'-deoxy-adenosine, cytidine, thymidine and guanosine nucleoside residues, respectively;

"A" is a 2'"'"'-methoxyethoxy adenosine nucleoside residue;

"T" is a 2'"'"'-methoxyethoxy thymidine nucleoside residue;

"G" is a 2'"'"'-methoxyethoxy guanosine nucleoside residue; and

"C" is a 2'"'"'-methoxyethoxy 5-methyl-cytidine nucleoside residue.

25. A pharmaceutical composition comprising the antisense oligonucleotide of claim 24 and a pharmaceutically acceptable carrier.

26. An antisense oligonucleotide comprising from about 8 to about 30 nucleotides connected by covalent linkages, wherein said antisense oligonucleotide comprises at least an 8 nucleobase portion of SEQ ID NO:

3, 11, 20 or 22, and wherein said antisense oligonucleotide encodes a murine intercellular adhesion molecule-1 and modulates the expression of the murine intercellular adhesion molecule-1.

27. An antisense oligonucleotide that modulates the expression of murine intercellular adhesion molecule-1, wherein said oligonucleotide is ISIS 17481 having the structure:

TsCsCsCsAsCsAsGsCsAsGsCsTsTsGsCsAsCsGsA,

wherein;

"s" is a phosphorothioate linkage;

"A", "T" and "G" are 2'"'"'-deoxy-adenosine, thymidine and guanosine nucleoside residues, respectively;

"C" is a 2'"'"'-deoxy-5-methyl-cytidine nucleoside residue;

"A" is a 2'"'"'-methoxyethoxy adenosine nucleoside residue;

"T" is a 2'"'"'-methoxyethoxy thymidine nucleoside residue;

"G" is a 2'"'"'-methoxyethoxy guanosine nucleoside residue; and

"C" is a 2'"'"'-methoxyethoxy 5-methyl-cytidine nucleoside residue.