IAP nucleobase oligomers and oligomeric complexes and uses thereof

• US 20050148535A1
• Filed: 10/28/2004
• Published: 07/07/2005
• Est. Priority Date: 10/30/2003
• Status: Abandoned Application

• Current Assignee: Aegera Therapeutics, Inc.
• Original Assignee: Aegera Therapeutics, Inc.
• Inventors: LaCasse, Eric, McManus, Daniel
• Application Number:
  US10/975,974
• Publication Number:
  US 20050148535A1
• Time in Patent Office: Days
• US Class Current: 514/44.A
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 35/02 specific for leukemia
• 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: C07H 21/04 with deoxyribosyl as saccharide radical
• 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 2310/14 interfering N.A.
• CPC Class Codes: C12N 2310/53 partially self-complementary or closed

Abstract

The present invention provides nucleobase oligomers and oligomer complexes that inhibit expression of an IAP polypeptide, and methods for using them to induce apoptosis in a cell. The nucleobase oligomers and oligomer complexes of the present invention may also be used to form pharmaceutical compositions. The invention also features methods for enhancing apoptosis in a cell by administering a nucleobase oligomer or oligomer complex of the invention in combination with a chemotherapeutic or chemosensitizing agent.

Claims

1. A substantially pure nucleobase oligomer comprising a duplex comprising at least eight but no more than thirty consecutive nucleobases corresponding to XIAP (SEQ ID NO:

21), HIAP-1 (SEQ ID NO;

     53), or HIAP-2 (SEQ ID NO;

     47), wherein said duplex is capable of reducing expression of an IAP polypeptide in a cell.

2. The oligomer of claim 1, wherein said duplex comprises a first domain comprising between 21 and 29 nucleobases and a second domain that hybridizes to said first domain under physiological conditions, wherein said first and second domains are connected by a single stranded loop.

3. The oligomer of claim 2, wherein said loop consists of between 6 and 12 nucleobases.

4. The oligomer of claim 2, wherein said loop consists of 8 nucleobases.

5. The oligomer of claim 1, wherein said duplex is selected from the group consisting of SEQ ID NOs:

32-36, and said oligomer reduces expression of XIAP.

6. The oligomer of claim 1, wherein said duplex is selected from the group consisting of SEQ ID NOs:

42-46, and said oligomer reduces expression of HIAP-2.

22. An expression vector encoding a nucleobase oligomer of claim 1 or nucleobase oligomeric complex of claim 7.

23. The expression vector of claim 22, wherein a nucleic acid sequence encoding said nucleobase oligomer or nucleobase oligomeric complex is operably linked to a promoter.

24. The expression vector of claim 23, wherein said promoter is the U6 PolIII promoter, or the polymerase III H1 promoter.

31. A method of enhancing apoptosis in a cell, said method comprising contacting said cell with an apoptosis-enhancing amount of a nucleobase oligomer of claim 1 or an oligomeric complex of claim 8.

32. The method of claim 31, wherein said cell is in vitro.

33. The method of claim 31, wherein said cell is in vivo.

34. The method of claim 31, wherein said cell is a human cell.

35. The method of claim 34, wherein said cell is a cancer cell

36. The method of claim 33, wherein said cell is from a human.

37. A method of enhancing apoptosis in a cell, said method comprising contacting said cell with (i) a nucleobase oligomer of claim 1 or an oligomeric complex of claim 8;

and (ii) a chemotherapeutic agent in amounts that together are sufficient to enhance apoptosis in said cell.

38. The method, according to claim 37, in which the chemotherapeutic agent is selected from the agents listed on Table 5.

39. A method of enhancing apoptosis in a cell, said method comprising contacting said cell with (i) a nucleobase oligomer of claim 1 or an oligomeric complex of claim 8;

and (ii) a chemosensitizing agent in amounts that together are sufficient to enhance apoptosis in said cell.

40. A method of treating a proliferative disease in an individual in need thereof, said method comprising administering to said individual a nucleobase oligomer of claim 1 or an oligomeric complex of claim 8 in an amount effective to treat said individual.

41. A method of treating a proliferative disease in an individual in need thereof, said method comprising administering to said individual (i) a nucleobase oligomer of claim 1 or an oligomeric complex of claim 8;

and (ii) a chemotherapeutic agent in amounts sufficient to treat said individual.

42. A method of treating a proliferative disease in an individual in need thereof, said method comprising administering to said individual (i) a nucleobase oligomer of claim 1 or an oligomeric complex of claim 8;

and (ii) a chemosensitizing agent in amounts sufficient to treat said individual.

43. The method of any one of claims 40-42, wherein said animal is a human.

44. The method of any one of claims 40-42, wherein said proliferative disorder is cancer.

45. The method of claim 44, wherein said cancer is selected from the group consisting of acute leukemia, acute lymphocytic leukemia, acute myelocytic leukemia, acute myeloblastic leukemia, acute promyelocytic leukemia, acute myelomonocytic leukemia, acute monocytic leukemia, acute erythroleukemia, chronic leukemia, chronic myelocytic leukemia, chronic lymphocytic leukemia), polycythemia vera, Hodgkin'"'"'s disease, non-Hodgkin'"'"'s disease, Waldenstrom'"'"'s macroglobulinemia, heavy chain disease, fibrosarcoma, myxosarcoma, liposarcoma, chondrosarcoma, osteogenic sarcoma, chordoma, angiosarcoma, endotheliosarcoma, lymphangiosarcoma, lymphangioendotheliosarcoma, synovioma, mesothelioma, Ewing'"'"'s tumor, leiomyosarcoma, rhabdomyosarcoma, colon carcinoma, pancreatic cancer, breast cancer, ovarian cancer, prostate cancer, squamous cell carcinoma, basal cell carcinoma, adenocarcinoma, sweat gland carcinoma, sebaceous gland carcinoma, papillary carcinoma, papillary adenocarcinomas, cystadenocarcinoma, medullary carcinoma, bronchogenic carcinoma, renal cell carcinoma, hepatoma, bile duct carcinoma, choriocarcinoma, seminoma, embryonal carcinoma, Wilm'"'"'s tumor, cervical cancer, uterine cancer, testicular cancer, lung carcinoma, small cell lung carcinoma, bladder carcinoma, epithelial carcinoma, glioma, astrocytoma, medulloblastoma, craniopharyngioma, ependymoma, pinealoma, hemangioblastoma, acoustic neuroma, oligodendroglioma, schwannoma, meningioma, melanoma, neuroblastoma, and retinoblastoma.

46. A kit comprising:

(i) a nucleobase oligomer of claim 1 or an oligomeric complex of claim 8;

and

(ii) instructions to administer said nucleobase oligomer or oligomeric complex to an individual having a proliferative disease in an amount sufficient to treat said proliferative disease.

47. A kit comprising:

(i) a nucleobase oligomer of claim 1 or an oligomeric complex of claim 8;

and

(ii) instructions to administer said nucleobase oligomer or oligomeric complex and a chemotherapeutic agent to an individual having a proliferative disease in amounts sufficient to treat said proliferative disease.

48. A kit comprising:

(i) a nucleobase oligomer of claim 1 or an oligomeric complex of claim 8;

and

(ii) instructions to administer said nucleobase oligomer or oligomeric complex and a chemosensitizing agent to an individual having a proliferative disease in amounts sufficient to treat said proliferative disease.

7. A nucleobase oligomeric complex comprising paired sense and antisense strands, wherein said complex comprises between eight and thirty consecutive nucleobases corresponding to a sequence of any one of XIAP (SEQ ID NO:

21), HIAP-1 (SEQ ID NO;

     53), and HIAP-2 (SEQ ID NO;

     47), and said complex is capable of reducing expression of an IAP polypeptide in a cell.

8. The complex of claim 7, wherein said complex comprises any one of SEQ ID NOs:

1-31.

9. The complex of claim 7, wherein said complex comprises any one of SEQ ID NOs:

37-41 and 54-63.

10. The complex of claim 8, wherein said complex is dsRNA.

11. The complex of claim 8, wherein said complex comprises at least one modification.

12. The complex of claim 11, wherein said complex comprises at least two modifications.

13. The complex of claim 11, wherein said modification is a modified sugar moiety, nucleobase, or internucleoside linkage.

14. The complex of claim 13, wherein said modification is a modified internucleoside linkage selected from the group consisting of phosphorothioate, methylphosphonate, phosphotriester, phosphorodithioate, and phosphoselenate linkages.

15. The complex of claim 13, wherein said modification is a modified sugar moiety.

16. The complex of claim 13, wherein said modified sugar moiety is a 2′

-O-methyl group or a 2′

-O-methoxyethyl group.

17. The complex of claim 13, wherein said modification is a modified nucleobase.

18. The complex of claim 17, wherein said modified nucleobase is 5-methyl cytosine.

19. The complex of claim 17, wherein said modified nucleobase is a chimeric nucleobase oligomer.

20. The complex of claim 19, wherein said chimeric nucleobase oligomer comprises a plurality of RNA residues.

21. The complex of claim 20, wherein said RNA residues are linked together by phosphorothioate linkages.

25. A cell comprising the expression vector of claim 21.

26. The cell of claim 25, wherein said cell is a transformed human cell that stably expresses said expression vector.

27. The cell of claim 25, wherein said cell is in vivo.

28. The cell of claim 27, wherein said cell is a human cell.

29. The cell of claim 28, wherein said cell is a neoplastic cell.

30. The cell of claim 27, wherein said cell is in a human.