Pharmaceutical compositions and methods useful for modulating angiogenesis, inhibiting metastasis and tumor fibrosis, and assessing the malignancy of colon cancer tumors

• US 20060127402A1
• Filed: 11/27/2003
• Published: 06/15/2006
• Est. Priority Date: 11/27/2002
• Status: Active Grant

• Current Assignee: TECHNION RESEARCH AND DEVELOPMENT FOUNDATION, LTD.
• Original Assignee: TECHNION RESEARCH AND DEVELOPMENT FOUNDATION, LTD.
• Inventors: Gengrinovitch, Stela, Akiri, Gal, Vadasz, Zahava, Neufeld, Gera
• Granted Patent: US 8,163,494 B2
• Time in Patent Office: Days
• US Class Current: 424/146.100
• CPC Class Codes: A61K 2039/505 comprising antibodies
• 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 35/04 specific for metastasis
• CPC Class Codes: A61P 9/00 Drugs for disorders of the cardiovascular system
• CPC Class Codes: C07K 16/30 from tumour cells
• CPC Class Codes: C07K 16/40 against enzymes
• CPC Class Codes: C12N 9/0022 with oxygen as acceptor (1.4.3)
• CPC Class Codes: C12Q 1/26 involving oxidoreductase
• CPC Class Codes: C12Q 1/6886 for cancer immunoassay for cancer G01N33/574
• CPC Class Codes: C12Y 104/03013 Protein-lysine 6-oxidase (1.4.3.13), i.e. lysyl-oxidase
• CPC Class Codes: G01N 2500/00 Screening for compounds of potential therapeutic value
• CPC Class Codes: G01N 33/574 for cancer
• CPC Class Codes: G01N 33/57419 of colon

Abstract

Methods and compositions suitable for modulating angiogenesis in a mammalian tissue are provided. Further provided are methods suitable for inhibiting metastasis and fibrosis in a mammalian tissue and for assessing the malignancy of colon cancer tumors.

Claims

1. A method of modulating angiogenesis in a mammalian tissue, the method comprising administering into the mammalian tissue a molecule capable of modifying a tissue level and/or activity of at least one type of lysyl oxidase to thereby modulate angiogenesis in the mammalian tissue.

2. The method of claim 1, wherein said molecule is an antibody or an antibody fragment capable of binding with, and at least partially inhibiting said activity of, said at least one polypeptide.

3. The method of claim 2, wherein said antibody or said antibody fragment is directed against at least an antigenic portion of the polypeptide set forth in SEQ ID NO:

2, 3, 6, 8 or 9.

4. The method of claim 1, wherein said molecule is a polynucleotide capable of downregulating expression of said at least one type of lysyl oxidase.

5. The method of claim 4, wherein said polynucleotide is at least partially complementary with the polynucleotide set forth in SEQ ID NO:

1, 4, 5 or 7.

6. The method of claim 1, wherein said molecule is a polypeptide having lysyl oxidase activity.

7. The method of claim 6, wherein said polypeptide is as set forth in SEQ ID NO:

2, 3, 6, 8 or 9.

8. A method of modulating angiogenesis in a mammalian tissue, the method comprising administering into the mammalian tissue a nucleic acid construct being capable of expressing a polypeptide having lysyl oxidase activity to thereby modulate angiogenesis within the mammalian tissue.

9. The method of claim 8, wherein said polypeptide is at least 75% homologous to the polypeptide set forth in SEQ ID NO:

2, 3, 6, 8 or 9.

10. A method of identifying molecules capable of modulating angiogenesis, the method comprising:

(a) isolating molecules which exhibit specific reactivity with at least one type of lysyl oxidase; and

(b) testing said molecules within mammalian tissue so as to determine the angiogenesis modulation activity thereof.

11. The method of claim 10, wherein step (a) is effected by binding assays and/or lysyl oxidase activity assays.

12. A pharmaceutical composition useful for modulating angiogenesis in mammalian tissue, the composition-of matter comprising, as an active ingredient, a molecule capable of modifying a level and/or activity of at least one type of lysyl oxidase of the mammalian tissue and a pharmaceutically effective carrier.

13. The pharmaceutical composition of claim 12, wherein said molecule is an antibody or an antibody fragment capable of binding with, and at least partially inhibiting said activity of, said at least one type of lysyl oxidase.

14. The pharmaceutical composition of claim 13, wherein said antibody or said antibody fragment is directed against at least a portion of the polypeptide set forth in SEQ ID NO:

2, 3, 6, 8 or 9.

15. The pharmaceutical composition of claim 12, wherein said molecule is a polynucleotide capable of downregulating expression of said at least one type of lysyl oxidase.

16. The pharmaceutical composition of claim 15, wherein said polynucleotide is at least partially complementary with the polynucleotide set forth in SEQ ID NO:

1, 4, 5 or 7.

17. The pharmaceutical composition of claim 12, wherein said molecule is a polypeptide having lysyl oxidase activity.

18. The pharmaceutical composition of claim 12, wherein said polypeptide is as set forth SEQ ID NO:

2, 3, 6, 8 or 9.

19. A method of modulating angiogenesis in a mammalian tissue, the method comprising administering into the mammalian tissue a molecule capable of modifying a tissue level and/or a lysyl oxidase activity of a polypeptide at least 75% homologous to the polypeptide set forth in SEQ ID NO:

2 to thereby modulate angiogenesis in the mammalian tissue.

20. A method of modulating angiogenesis in a mammalian tissue, the method comprising administering into the mammalian tissue a nucleic acid construct being capable of expressing a polypeptide at least 75% homologous to the polypeptide set forth in SEQ ID NO:

2 to thereby modulate angiogenesis within the mammalian tissue.

21. A method of determining the malignancy of cancerous tissue, the method comprising

(a) determining a lysyl oxidase expression level and/or activity of the cancerous tissue;

and

(b) comparing said lysyl oxidase expression level and/or activity with that determined for control tissue to thereby determine the malignancy of the cancerous tissue.

22. A method of inhibiting metastasis and/or fibrosis in a mammalian tissue, the method comprising administering to the mammalian tissue a molecule capable of downregulating a tissue level and/or activity of at least one type of lysyl oxidase to thereby inhibit metastasis in the mammalian tissue.

23. The method of claim 22, wherein said molecule is an antibody or an antibody fragment capable of specifically binding with said at least one type of lysyl oxidase.

24. The method of claim 22, wherein said antibody or said antibody fragment is directed against at least an antigenic portion of the polypeptide set forth in SEQ ID NO:

2, 3, 6, 8 or 9.

25. The method of claim 22, wherein said molecule is a polynucleotide capable of downregulating expression of said at least one type of lysyl oxidase.

26. The method of claim 25, wherein said polynucleotide is at least partially complementary with the polynucleotide set forth in SEQ ID NO:

1, 4, 5 or 7.

27. A method of identifying molecules capable of inhibiting metastasis and/or fibrosis, the method comprising:

(a) screening and identifying molecules which exhibit specific reactivity with at least one type of lysyl oxidase; and

(b) testing a metastasis and/or fibrosis inhibitory potential of said molecules.

28. The method of claim 27, wherein step (a) is effected by binding assays and/or lysyl oxidase activity assays.

29. The method of claim 27, wherein step (b) is effected in-vivo or in-vitro.

30. A pharmaceutical composition useful for inhibiting metastasis and/or fibrosis in mammalian tissue, comprising, as an active ingredient, a molecule capable of downregulating a level and/or activity of at least one type of lysyl oxidase present within the mammalian tissue and a pharmaceutically effective carrier.

31. The pharmaceutical composition of claim 30, wherein said molecule is an antibody or an antibody fragment capable of specifically binding with said at least one type of lysyl oxidase.

32. The pharmaceutical composition of claim 31, wherein said antibody or said antibody fragment is directed against at least an antigenic portion of the polypeptide set forth in SEQ ID NO:

2, 3, 6, 8 or 9.

33. The pharmaceutical composition of claim 30, wherein said molecule is a polynucleotide capable of downregulating expression of said at least one type of lysyl oxidase.

34. The pharmaceutical composition of claim 33, wherein said polynucleotide is at least partially complementary with the polynucleotide set forth in SEQ ID NO:

1, 4, 5 or 7.

35. A method of inhibiting metastasis and fibrosis in a mammalian tissue, the method comprising administering to the mammalian tissue a molecule capable of downregulating a tissue level and/or a lysyl oxidase activity of a polypeptide at least 75% homologous to the polypeptide set forth in SEQ ID NO:

2 or 9, to thereby inhibit metastasis and fibrosis in the mammalian tissue.

36. A method of assessing a malignancy of a colon tumor comprising determining a tissue level and/or an activity level of a polypeptide at least 75% homologous to the polypeptide set forth in SEQ ID NO:

2 or 9 in a tissue of the colon tumor, thereby assessing the malignancy of the colon tumor.

37. The method of claim 36, wherein said tissue of said colon tumor is obtained using a colon biopsy and/or a colon surgery.

38. The method of claim 36, wherein said determining said tissue level of said polypeptide is effected by an immunological detection method and/or an RNA detection method.

39. The method of claim 36, wherein said determining said activity level of said polypeptide is effected by an enzymatic activity detection method.

40. The method of claim 38, wherein said immunological detection method is selected from the group consisting of a radio-immunoassay (RIA), an enzyme linked immunosorbent assay (ELISA), a Western blot analysis, and an immunohistochemical analysis.

41. The method of claim 38, wherein said RNA detection method is selected from the group consisting of a Northern blot analysis, an RNA in situ hybridization stain, an RT-PCR analysis, and an in situ RT-PCR stain.

42. The method of claim 39, wherein said enzymatic activity detection method is selected from the group consisting of a cytochemical stain, an in vitro activity assay, and an activity gel.

43. The method of claim 36, wherein the malignancy of the colon tumor is assessed by comparing said tissue level and/or said activity level of said polypeptide in said tissue of said colon tumor with a tissue level and/or activity level of said polypeptide in a normal colon tissue.

44. A method of predicting a prognosis of an individual diagnosed with colon cancer comprising:

(a) obtaining a colon tumor tissue from the individual, and;

(b) determining a tissue level and/or an activity level of a polypeptide at least 75% homologous to the polypeptide set forth in SEQ ID NO;

2 or 9 in said colon tumor tissue to thereby assess the malignancy of said colon tumor tissue and predict the prognosis of the individual diagnosed with colon cancer.

45. The method of claim 44, wherein said obtaining is effected using a colon biopsy and/or a colon surgery.

46. The method of claim 44, wherein said determining said tissue level of said polypeptide is effected by an immunological detection method and/or an RNA detection method.

47. The method of claim 44, wherein said determining said activity level of said polypeptide is effected by an enzymatic activity detection method.

48. The method of claim 46, wherein said immunological detection method is selected from the group consisting of a radio-immunoassay (RIA), an enzyme linked immunosorbent assay (ELISA), a Western blot analysis, and an immunohistochemical analysis.

49. The method of claim 46, wherein said RNA detection method is selected from the group consisting of a Northern blot analysis, an in situ hybridization stain, an RT-PCR analysis, and an in situ RT-PCR stain.

50. The method of claim 47, wherein said enzymatic activity detection method is selected from the group consisting of a cytochemical stain, an in vitro activity assay, and an activity gel.

51. The method of claim 44, wherein the malignancy of said colon tumor tissue is assessed by comparing said tissue level and/or said activity level of said polypeptide in said colon tumor tissue with a tissue level and/or activity level of said polypeptide in a normal colon tissue.