Antisense modulation of Smad6 expression

• US 6,566,131 B1
• Filed: 10/04/2000
• Issued: 05/20/2003
• Est. Priority Date: 10/04/2000
• Status: Expired due to Fees

• Current Assignee: ISIS Pharmaceuticals Incorporated
• Original Assignee: ISIS Pharmaceuticals Incorporated
• Inventors: Cowsert, Lex M.
• Primary Examiner(s): McGarry; Sean
• Assistant Examiner(s): SCHMIDT, MARY M
• Application Number:
  US09/679,298
• Time in Patent Office: 958 Days
• Field of Search: 536/23.1,24.1,24.5,24.31,24.33 435/320.1,6,91.1,325,366,375 514/44
• US Class Current: 435/375
• 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: 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 2310/315 Phosphorothioates
• CPC Class Codes: C12N 2310/321 2'-O-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 2310/3525 MOE, methoxyethoxy
• CPC Class Codes: Y02P 20/582 Recycling of unreacted starting or intermediate materials

Abstract

Antisense compounds, compositions and methods are provided for modulating the expression of Smad6. The compositions comprise antisense compounds, particularly antisense oligonucleotides, targeted to nucleic acids encoding Smad6. Methods of using these compounds for modulation of Smad6 expression and for treatment of diseases associated with expression of Smad6 are provided.

Claims

1. A compound 8 to 50 nucleobases in length targeted to nucleobases 40 through 57, nucleobases 99 through 116, nucleobases 156 through 173, nucleobases 180 through 197, nucleobases 219 through 236, nucleobases 286 through 303, nucleobases 411 through 428, nucleobases 537 through 554, or nucleobases 786 through 803 of a 5′

-untranslated region, nucleobases 948 through 965, nucleobases 1014 through 1031, nucleobases 1033 through 1050, nucleobases 1080 through 1097, nucleobases 1277 through 1294, nucleobases 1314 through 1331, nucleobases 1427 through 1444, nucleobases 1456 through 1473, nucleobases 1729 through 1746, nucleobases 1945 through 1962, nucleobases 2001 through 2018, nucleobases 2066 through 2083, nucleobases 2162 through 2179, or nucleobases 2326 through 2343 of a coding region of a nucleic acid molecule encoding human Smad6 of SEQ ID NO;

3, wherein said compound specifically hybridizes with one of said regions and inhibits the expression of human Smad6.

2. The compound of claim 1 which is an antisense oligonucleotide.

4. The compound of claim 2 wherein the antisense oligonucleotide has a sequence comprising SEQ ID NO:

26, 29 or 40.

5. The compound of claim 2 wherein the antisense oligonucleotide comprises at least one modified internucleoside linkage.

6. The compound of claim 5 wherein the modified internucleoside linkage is a phosphorothioate linkage.

7. The compound of claim 2 wherein the antisense oligonucleotide comprises at least one modified sugar moiety.

8. The compound of claim 7 wherein the modified sugar moiety is a 2′

-O-methoxyethyl sugar moiety.

9. The compound of claim 2 wherein the antisense oligonucleotide comprises at least one modified nucleobase.

10. The compound of claim 9 wherein the modified nucleobase is a 5-methylcytosine.

11. The compound of claim 2 wherein the antisense oligonucleotide is a chimeric oligonucleotide.

12. A composition comprising the compound of claim 1 and a pharmaceutically acceptable carrier or diluent.

13. The composition of claim 12 further comprising a colloidal dispersion system.

14. The composition of claim 12 wherein the compound is an antisense oligonucleotide.

15. A method of inhibiting the expression of human Smad6 in cells or tissues comprising contacting said cells or tissues in vitro with the compound of claim 1 so that expression of human Smad6 is inhibited.

3. A compound up to 50 nucleobases in length comprising at least an 8-nucleobase portion of SEQ ID NO:

12, 13, 19, 26, 27, 29, 40, 11, 14, 17, 21, 28, 41 or 45 which inhibits the expression of human Smad6.

16. The compound of claim 3 which is an antisense oligonucleotide.

17. The compound of claim 16 wherein the antisense oligonucleotide comprises at least one modified internucleoside linkage.

18. The compound of claim 17 wherein the modified internucleoside linkage is a phosphorothioate linkage.

19. The compound of claim 16 wherein the antisense oligonucleotide comprises at least one modified sugar moiety.

20. The compound of claim 19 wherein the modified sugar moiety is a 2′

-O-methoxyethyl sugar moiety.

21. The compound of claim 16 wherein the antisense oligonucleotide comprises at least one modified nucleobase.

22. The compound of claim 21 wherein the modified nucleobase is a 5-methylcytosine.

23. The compound of claim 16 wherein the antisense oligonucleotide is a chimeric oligonucleotide.

24. A method of inhibiting the expression of human Smad6 in human cells or tissues comprising contacting said cells or tissues in vitro with the compound of claim 3 so that expression of human Smad6 is inhibited.

25. A composition comprising the compound of claim 3 and a pharmaceutically acceptable carrier or diluent.

26. The composition of claim 25 further comprising a colloidal dispersion system.

27. The composition of claim 25 wherein the compound is an