Antisense modulation of bcl-x expression

• US 20010007025A1
• Filed: 12/12/2000
• Published: 07/05/2001
• Est. Priority Date: 10/07/1998
• Status: Abandoned Application

• Current Assignee: ISIS Pharmaceuticals Incorporated (Ionis Pharmaceuticals, Inc.)
• Original Assignee: ISIS Pharmaceuticals Incorporated (Ionis Pharmaceuticals, Inc.)
• Inventors: Monia, Brett P., Dean, Nicholas M., Bennett, C. Frank, Nickoloff, Brian J., Zhang, Qing Qing
• Application Number:
  US09/734,846
• Publication Number:
  US 20010007025A1
• Time in Patent Office: Days
• 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: 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: C12N 15/1135 against oncogenes or tumor suppressor genes
• CPC Class Codes: C12N 2310/11 Antisense
• CPC Class Codes: C12N 2310/315 Phosphorothioates
• CPC Class Codes: C12N 2310/3181 Peptide nucleic acid, PNA
• CPC Class Codes: C12N 2310/321 2'-O-R Modification
• CPC Class Codes: C12N 2310/3233 Morpholino-type ring
• CPC Class Codes: C12N 2310/334 Modified C
• 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

Abstract

Compositions and methods are provided for modulating the expression of bcl-x. Antisense compounds, particularly antisense oligonucleotides, targeted to nucleic acids encoding bcl-x are preferred. Methods of using these compounds for modulation of bcl-x expression and for treatment of diseases associated with expression of bcl-x are also provided. Methods of sensitizing cells to apoptotic stimuli are also provided.

Claims

1. An antisense compound 8 to 30 nucleobases in length targeted to a nucleic acid molecule encoding a human bcl-x, wherein said antisense compound modulates the expression of human bcl-x.

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, a morpholino linkage or a peptide-nucleic acid linkage.

5. The antisense compound of

claim 1

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 or a 2′

-dimethylaminooxyethoxy sugar moiety.

7. The antisense compound of

claim 5

wherein substantially all sugar moieties of the antisense oligonucleotide are modified sugar moieties.

8. The antisense compound of

claim 3

wherein the antisense oligonucleotide comprises at least one modified nucleobase.

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 8

wherein each 2′

-O-methoxyethyl modified cytosine nucleobase of the antisense oligonucleotide is a 5-methylcytosine.

11. The antisense compound of

claim 1

which is a chimeric oligonucleotide.

12. A pharmaceutical composition comprising the antisense compound of

claim 1

and a pharmaceutically acceptable carrier or diluent.

13. The pharmaceutical composition of

claim 12

further comprising a colloidal dispersion system.

14. The pharmaceutical composition of

claim 12

wherein the antisense compound is an antisense oligonucleotide.

15. The antisense compound of

claim 1

which is targeted to bcl-xl.

16. The antisense compound of

claim 1

which is targeted to a nucleic acid molecule encoding bcl-xl and which preferentially inhibits the expression of bcl-xl.

17. The antisense compound of

claim 16

which is targeted to a region of a nucleic acid molecule encoding bcl-xl which is not found in a nucleic acid molecule encoding bcl-xs.

18. The antisense compound of

claim 16

which promotes apoptosis.

19. The antisense compound of

claim 1

which is targeted to a region of a nucleic acid molecule encoding bcl-xs and which reduces the expression of bcl-xs.

20. The antisense compound of

claim 19

which inhibits apoptosis.

21. The antisense compound of

claim 1

which alters the ratio of bcl-x isoforms expressed by a cell or tissue.

22. The antisense compound of

claim 21

which increases the ratio of bcl-xl to bcl-xs expressed.

23. The antisense compound of

claim 21

which decreases the ratio of bcl-xl to bcl-xs expressed.

24. A method of inhibiting the expression of bcl-x in human cells or tissues comprising contacting said cells or tissues with the antisense compound of

claim 1

so that expression of bcl-x is inhibited.

25. A method of treating an animal having a disease or condition associated with bcl-x comprising administering to said animal a therapeutically or prophylactically effective amount of the antisense compound of

claim 1

so that expression of bcl-x is inhibited.

26. A method of treating an animal having a disease or condition characterized by a reduction in apoptosis comprising administering to said human a prophylactically or therapeutically effective amount of the antisense compound of

claim 1

.

27. The method of

claim 26

wherein the antisense compound is targeted to a nucleic acid molecule encoding bcl-xl and which preferentially inhibits the expression of bcl-xl.

28. The pharmaceutical composition of

claim 12

further comprising a chemotherapeutic agent for the treatment of cancer.

29. A method of treating cancer in an animal comprising:

(a) administering to the animal a pharmaceutical composition of

claim 12

; and

(b) administering to the animal a chemotherapeutic agent for the treatment of cancer.

30. A method of sensitizing a cell to an apoptotic stimulus comprising treating the cell with the composition of

claim 1

.

31. The method of

claim 30

wherein the apoptotic stimulus is radiation.

32. The method of

claim 31

wherein the radiation is ultraviolet radiation.

33. The method of

claim 30

wherein the apoptotic stimulus is a cancer chemotherapeutic drug.

34. The method of

claim 33

wherein the cancer chemotherapeutic drug is VP-16, cisplatinum or taxol.

35. The method of

claim 30

wherein the apoptotic stimulus is a cellular signaling molecule.

36. The method of

claim 30

wherein the apoptotic stimulus is ceramide, a cytokine or staurosporine.

37. The method of

claim 30

wherein said apoptotic stimulus causes mitochondrial dysfunction.

38. The method of

claim 37

wherein said mitochondrial dysfunction is loss of mitochondrial membrane potential.

39. The method of

claim 30

, wherein said cell is a cancer cell.

40. The method of

claim 39

, wherein said cancer cells are glioblastoma or leukemia cells.

41. A method of promoting apoptosis of cancer cells, comprising contacting said cells with the antisense compound of

claim 1

.

42. The method of

claim 41

, further comprising the step of contacting said cells with a chemotherapeutic agent.

43. The method of

claim 42

, wherein said chemotherapeutic agent is doxorubicin or dexamethasone.

44. The method of

claim 42

, wherein said cancer cells are glioblastoma cells or leukemia cells.