Antisense modulation of novel anti-apoptotic bcl-2-related proteins

• US 6,001,992 A
• Filed: 01/07/1999
• Issued: 12/14/1999
• Est. Priority Date: 01/07/1999
• Status: Expired due to Term

• Current Assignee: ISIS Pharmaceuticals Incorporated (Ionis Pharmaceuticals, Inc.)
• Original Assignee: ISIS Pharmaceuticals Incorporated (Ionis Pharmaceuticals, Inc.)
• Inventors: Marcusson, Eric G., Dean, Nicholas M., Bennett, C. Frank, Ackermann, Elizabeth J.
• Primary Examiner(s): Degen, Nancy
• Assistant Examiner(s): EPPS -SMITH, JANET L
• Application Number:
  US09/226,568
• Time in Patent Office: 341 Days
• Field of Search: 435/325, 435/6, 435/91.1, 435/440, 536/24.5, 536/24.33, 536/24.3, 536/23.1, 514/44
• US Class Current: 536/24.5
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• 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 15/1135 against oncogenes or tumor suppressor genes
• CPC Class Codes: C12N 2310/315 Phosphorothioates
• CPC Class Codes: C12N 2310/318 where the PO2 is completely replaced, e.g. MMI or formacetal
• CPC Class Codes: C12N 2310/3181 Peptide nucleic acid, PNA
• 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 2310/345 having at least two different backbone modifications
• 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

Compositions and methods are provided for modulating the expression of novel anti-apoptotic bcl-2-related proteins. Antisense oligonucleotides targeted to nucleic acids encoding the human novel anti-apoptotic bcl-2-related proteins A1 and mcl-1 are preferred. Methods of using these compounds for modulation of novel anti-apoptotic bcl-2-related protein expression and for treatment of diseases associated with expression of novel anti-apoptotic bcl-2-related proteins are also provided. Also provided are methods of using these compounds for promoting apoptosis and for treatment of diseases for which promotion of apoptosis is desired.

Claims

1. An antisense compound 8 to 30 nucleobases in length targeted to either SEQ ID NO:

1 or SEQ ID NO;

18, nucleic acid molecules encoding novel anti-apoptotic bcl-2-related proteins, wherein said antisense compound inhibits the expression of said novel anti-apoptotic bcl-2-related proteins.

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

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

4. The antisense compound of claim 3 wherein the modified internucleoside linkage of the antisense oligonucleotide is a phosphorothioate linkage.

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

6. The antisense compound of claim 5 wherein the modified sugar moiety of the antisense oligonucleotide is a 2'"'"'-O-methoxyethyl sugar moiety.

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

8. The antisense compound of claim 7 wherein the modified nucleobase is a 2'"'"'-O-methoxyethyl modified cytosine.

9. The antisense compound of claim 8 wherein the modified nucleobase of the antisense oligonucleotide is a 5-methylcytosine.

10. The antisense compound of claim 1 which is a chimeric oligonucleotide.

11. The antisense compound of claim 1 which is targeted to a nucleic acid encoding human A1.

12. The antisense compound of claim 1 which is targeted to a nucleic acid encoding human mcl-1.

13. A composition comprising the antisense compound of claim 1 and a carrier or diluent.

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

15. A method of inhibiting the expression of a novel human anti-apoptotic bcl-2-related protein in human cells or tissues comprising contacting said cells or tissues in vitro with the antisense compound of claim 1 so that expression of said novel human anti-apoptotic bcl-2-related protein is inhibited.

16. The method of claim 15 wherein the antisense compound is targeted to a nucleic acid molecule encoding human A1.

17. The method of claim 15 wherein the antisense compound is targeted to a nucleic acid molecule encoding human mcl-1.

18. A method of promoting apoptosis in human cells or tissues comprising contacting said cells or tissues in vitro with the antisense compound of claim 1 so that expression of said novel human anti-apoptotic bcl-2-related protein is inhibited.

19. The method of claim 18 wherein the antisense compound is targeted to a nucleic acid molecule encoding human A1.

20. The method of claim 18 wherein the antisense compound is targeted to a nucleic acid molecule encoding human mcl-1.

21. An antisense compound up to 30 nucleobases in length targeted to a nucleic acid encoding human A1 wherein the antisense compound comprises at least 8 nucleobases of SEQ ID NO:

3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15 or 16, and inhibits expression of said human A1.

22. The antisense compound of claim 21 which is an antisense oligonucleotide.

23. The antisense compound of claim 22 wherein the antisense oligonucleotide comprises at least one modified internucleoside linkage.

24. The antisense compound of claim 23 wherein the modified internucleoside linkage of the antisense oligonucleotide is a phosphorothioate linkage.

25. The antisense compound of claim 22 wherein the antisense oligonucleotide comprises at least one modified sugar moiety.

26. The antisense compound of claim 25 wherein the modified sugar moiety of the antisense oligonucleotide is a 2'"'"'-O-methoxyethyl sugar moiety.

27. The antisense compound of claim 22 wherein the antisense oligonucleotide comprises at least one modified nucleobase.

28. The antisense compound of claim 27 wherein the modified nucleobase is a 2'"'"'-O-methoxyethyl modified cytosine.

29. The antisense compound of claim 28 wherein the modified nucleobase of the antisense oligonucleotide is a 5'"'"'-methylcytosine.

30. The antisense compound of claim 21 which is a chimeric oligonucleotide.

31. An antisense compound up to 30 nucleobases in length targeted to a nucleic acid encoding human mcl-1 wherein the antisense compound comprises at least 8 nucleobases of SEQ ID NO:

22, 23, 31, 35, or 36, and inhibits expression of said human mcl-1.

32. The antisense compound of claim 31 which is an antisense oligonucleotide.

33. The antisense compound of claim 32 wherein the antisense oligonucleotide comprises at least one modified internucleoside linkage.

34. The antisense compound of claim 33 wherein the modified internucleoside linkage of the antisense oligonucleotide is a phosphorothioate linkage.

35. The antisense compound of claim 32 wherein the antisense oligonucleotide comprises at least one modified sugar moiety.

36. The antisense compound of claim 35 wherein the modified sugar moiety of the antisense oligonucleotide is a 2'"'"'-O-methoxyethyl sugar moiety.

37. The antisense compound of claim 32 wherein the antisense oligonucleotide comprises at least one modified nucleobase.

38. The antisense compound of claim 37 wherein the modified nucleobase is a 2'"'"'-O-methoxyethyl modified cytosine.

39. The antisense compound of claim 38 wherein the modified nucleobase of the antisense oligonucleotide is a 5'"'"'-methylcytosine.

40. The antisense compound of claim 31 which is a chimeric oligonucleotide.