Cancer associated araf1 protein kinase and its uses

• US 20060099142A1
• Filed: 03/13/2003
• Published: 05/11/2006
• Est. Priority Date: 03/14/2002
• Status: Abandoned Application

• Current Assignee: QT Incorporated
• Original Assignee: QT Incorporated
• Inventors: Delaney, Allen D.
• Application Number:
  US10/508,100
• Publication Number:
  US 20060099142A1
• Time in Patent Office: Days
• US Class Current: 424/1.49
• CPC Class Codes: A61K 2039/505 comprising antibodies
• CPC Class Codes: A61K 49/0004 Screening or testing of compounds for diagnosis of disorders, assessment of conditions, e.g. renal clearance, gastric emptying, testing for diabetes, allergy, rheuma, pancreas functions
• CPC Class Codes: A61K 49/0008 Screening agents using (non-human) animal models or transgenic animal models or chimeric hosts, e.g. Alzheimer disease animal model, transgenic model for heart failure
• CPC Class Codes: C12Q 1/485 involving kinase
• CPC Class Codes: C12Q 1/6886 for cancer immunoassay for cancer G01N33/574
• CPC Class Codes: C12Q 2600/136 Screening for pharmacological compounds
• CPC Class Codes: C12Q 2600/158 Expression markers
• CPC Class Codes: G01N 33/5011 for testing antineoplastic activity
• CPC Class Codes: G01N 33/57407 Specifically defined cancers
• CPC Class Codes: G01N 33/57492 involving compounds localized on the membrane of tumor or cancer cells

Abstract

Detection of expression of the provided protein kinase in cancers is useful as a diagnostic, for determining the effectiveness of drugs, and determining patient prognosis. The encoded polypeptides further provides a target for screening pharmaceutical agents effective in inhibiting the growth or metastasis of tumor cells. The present invention further provides methods and compositions relating to agents that specifically bind to Araf1 for treatment and visualization of tumors in patients.

Claims

1. A method of screening for biologically active agents that modulate a cancer associated protein kinase function, the method comprising:

combining a candidate biologically active agent with any one of;

(a) a polypeptide encoded by SEQ ID NO;

1;

or having the amino acid sequence set forth in SEQ ID NO;

2;

(b) a cell comprising a nucleic acid encoding a polypeptide encoded by SEQ ID NO;

1;

or

(c) a non-human transgenic animal model for cancer associated kinase gene function comprising one of;

(i) a knockout of a gene corresponding to SEQ ID NO;

1;

(ii) an exogenous and stably transmitted mammalian gene sequence comprising polypeptide encoded by SEQ ID NO;

1; and

determining the effect of said agent on kinase function.

2. A method for the diagnosis of cancer, the method comprising:

determining the upregulation of expression in SEQ ID NO;

1 in said cancer.

3. The method of claim 2, wherein said cancer is of the liver.

4. The method of claim 2, wherein said cancer is of the colon.

5. The method of claim 2, wherein said cancer is of the brain.

6. The method of claim 2, wherein said cancer is of the lung.

7. A method for inhibiting the growth of a cancer cell, the method comprising downregulating activity of the polypeptide encoded by SEQ ID NO:

1;

or having the amino acid sequence set forth in SEQ ID NO;

2;

in said cancer cell.

8. The method according to claim 7, wherein said method comprises introducing antisense sequences specific for SEQ ID NO:

1.

9. The method according to claim 7, wherein said method comprises introducing an inhibitor of kinase activity into said cancer cell.

10. The method according to claim 7, wherein said cancer cell is a liver cancer cell.

11. The method according to claim 7, wherein said cancer cell is a colon cancer cell.

12. The method according to claim 7, wherein said cancer cell is a brain cancer cell.

13. The method according to claim 7, wherein said cancer cell is a lung cancer cell.

14. A method of screening for targets of a cancer associated protein kinase, wherein said targets are associated with signal transduction in cancer cells, the method comprising:

comparing the pattern of gene expression in a normal cell, and in a tumour cell characterized by up-regulation of SEQ ID NO;

1.

15. The method according to claim 14, wherein said comparing the pattern of gene expression comprises quantitating specific mRNAs by hybridization to an array of polynucleotide probes.

16. A method of screening for targets of a cancer associated protein kinase, wherein said targets are associated with signal transduction in cancer cells, the method comprising:

comparing the pattern of protein phosphorylation in a normal cell, and in a tumour cell characterized by up-regulation of SEQ ID NO;

1.

17. A method to treat a tumour comprising administering a therapeutic amount of a composition comprising:

a compound of the general formula α

(P_z)C, wherein α

(P_z) is one or more moieties which specifically binds to a human protein Araf1, and C is one or more cytotoxic moieties; and

a pharmaceutically acceptable carrier.

18. The method of claim 17 wherein the tumour is a colon tumour.

19. The method of claim 17 wherein the tumour is a lung tumour.

20. The method of claim 17 wherein the tumour is a brain tumour.

21. The method of claim 17 wherein α

(P_z) is selected from the group consisting of an antibody and an antibody fragment.

22. The method of claim 21 wherein the antibody is selected from the group consisting of monoclonal antibodies, polyclonal antibodies, humanized antibodies, recombinant antibodies, chemically modified antibodies, and synthetic antibody analogs.

23. The method of claim 22 wherein C is a radioactive moiety.

24. The method of claim 23 wherein the radioactive moiety comprises a pharmaceutically acceptable radioactive isotope selected from the group consisting of ¹²³I, ¹²⁵I, ¹³¹I, ⁹⁰Y, ²¹¹At, ⁶⁷Cu, ¹⁸⁶Re, ¹⁸⁸Re, ²¹²Pb, and ²¹²Bi.

25. The method of claim 17 wherein C is a chemotoxic moiety.

26. The method of claim 17 wherein C is a toxin protein moiety.

27. The method of claim 26 wherein the toxin protein moiety is selected from the group consisting of ricin, abrin, diphtheria toxin, cholera toxin, gelonin, Pseudomonas exotoxin, Shigella toxin, and pokeweed antiviral protein.

28. A method for treating a tumour comprising administering a therapeutic amount of a composition comprising:

a compound of the general formula α

(P_z), wherein α

(P_z) is one or more moieties which specifically binds to a human protein Araf1, wherein the binding of α

(P_z) alters the function of Araf1, and a pharmaceutically acceptable carrier.

29. The method of claim 28 wherein the tumour is a liver tumour.

30. The method of claim 28 wherein the tumour is a colon tumour.

31. The method of claim 28 wherein the tumour is a brain tumour.

32. The method of claim 28 wherein the tumour is a lung tumour.

33. The method of claim 28 wherein α

(P_z) is selected from the group consisting of an antibody and an antibody fragment.

34. A composition for the treatment of a tumour comprising:

a compound of the general formula α

(P_z), wherein α

(P_z) is one or more moieties which specifically binds to a human Araf1, wherein the binding of α

(P_z) alters the function of protein Araf1, and a pharmaceutically acceptable carrier.

35. The composition of claim 34 wherein α

(P_z) is selected from the group consisting of an antibody and an antibody fragment

36. A method for visualizing a tumour in a patient, the method comprising:

a) administering to a patient an effective amount of a composition comprising;

a compound of the general formula α

(P_z)I, wherein α

(_z) is one or more moieties which specifically binds to a human Araf, and I is one or more imaging moieties; and

a pharmaceutically acceptable carrier; and

b) visualizing the imaging moieties of the compound.

37. The method of claim 36 wherein the tumour is a liver tumour.

38. The method of claim 36 wherein the tumour is a colon tumour.

39. The method of claim 36 wherein the tumour is a brain tumour.

40. The method of claim 36 wherein the tumour is a lung tumour.

41. The method of claim 36 wherein α

(P_z) is selected from the group consisting of an antibody and an antibody fragment.

42. The method of claim 36 wherein I is a radiographic moiety.

43. The method of claim 36 wherein I is a positron-emitting moiety.

44. The method of claim 36 wherein I is a magnetic spin contrast moiety.

45. The method of claim 44 wherein the magnetic spin contrast moiety comprises an ion selected from the group consisting of chromium(III), manganese(II), iron(II), nickel(II), copper(II), praseodymium(III), neodymium(III), samarium(III) and ytterbium(III).

46. The method of claim 36 wherein I is selected from the group consisting of an optically visible dye and an optically visible particle.

47-54. -54. (canceled)