Peptide tumor cell growth inhibitors

• US 6,355,619 B1
• Filed: 08/29/2000
• Issued: 03/12/2002
• Est. Priority Date: 06/14/1996
• Status: Expired due to Fees

• Current Assignee: Board of Regents of the University of Texas System (University of Texas System)
• Original Assignee: Board of Regents of the University of Texas System (University of Texas System)
• Inventors: Hayashi, Shinichiro, Miller, Edmund J.
• Primary Examiner(s): DAVENPORT, AVIS MICHELLE
• Application Number:
  US09/650,438
• Time in Patent Office: 560 Days
• Field of Search: 514/13, 514/14, 514/15, 514/16, 514/17, 514/18, 514/19, 530/300, 530/326, 530/327, 530/328, 530/329, 424/185.1
• US Class Current: 514/1.7
• CPC Class Codes: A61K 38/08 Peptides having 5 to 11 amino acids A61K38/043 - A61K38/046 take precedence
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: C07K 14/54 Interleukins [IL]
• CPC Class Codes: C07K 14/5421 IL-8
• CPC Class Codes: C07K 7/06 having 5 to 11 amino acids

Abstract

Disclosed are peptide-based compositions and methods for inhibiting and modulating the actions of CXC intercrine molecules. The antileukinate peptides described inhibit IL-8, GRO and MIP2β binding to neutrophils and neutrophil activation. The peptides are particularly advantageous as they inhibit IL-8-induced enzyme release at a 25 fold lower concentration than is required to inhibit chemotaxis, which makes them ideal for treating various inflammatory diseases and disorders including, amongst others, Adult Respiratory Distress Syndrome (ARDS), cystic fibrosis and chronic bronchitis. The invention further includes methods for inhibiting tumor cell growth by employing selected members of the disclosed group of peptides to inhibit a-chemokine binding to the tumor cell.

Claims

1. A method of inhibiting binding of an α

-chemokine to cognate α

-chemokine receptors, comprising contacting target cells that comprise cognate α

-chemokine receptors with a composition comprising at least one peptide that comprises the amino acid sequence Arg Arg X Trp Cys X (SEQ ID NO;

58), or a biologically functional equivalents thereof, wherein X is any amino acid, in an amount effective to inhibit binding of the α

-chemokine to its cognate receptors.

2. The method of claim 1 wherein the α

-chemokine is IL-8, MIP-2β

, MGSA/Groα

, PBP, IP10, PF4 or ENA78.

3. The method of claim 1 wherein the α

-chemokine is MGSA/Groα

.

4. The method of claim 1, wherein said α

-chemokine is IL-8.

5. The method of claim 1, wherein said at least one peptide comprises the amino acid sequence Arg Arg Trp Trp Cys Xaa₁ (SEQ ID NO:

23), wherein Xaa₁ is any amino acid.

6. The method of claim 5, wherein the at least one peptide comprises the amino acid sequence Arg Arg Trp Trp Cys Arg (SEQ ID NO:

1), Arg Arg Trp Trp Cys Ala (SEQ ID NO;

24), Arg Arg Trp Trp Cys Cys (SEQ ID NO;

25), Arg Arg Trp Trp Cys Asp (SEQ ID NO;

26), Arg Arg Trp Trp Cys Glu (SEQ ID NO;

27), Arg Arg Trp Trp Cys Phe (SEQ ID NO;

28), Arg Arg Trp Trp Cys Gly (SEQ ID NO;

29), Arg Arg Trp Tip Cys His (SEQ ID NO;

30), Arg Arg Trp Trp Cys Ile (SEQ ID NO;

31), Arg Arg Trp Trp Cys Lys (SEQ ID NO;

32), Arg Arg Trp Trp Cys Leu (SEQ ID NO;

33), Arg Arg Trp Trp Cys Met (SEQ ID NO;

34), Arg Arg Trp Trp Cys Asn (SEQ ID NO;

35), Arg Arg Trp Trp Cys Pro (SEQ ID NO;

36), Arg Arg Trp Trp Cys Gln (SEQ ID NO;

37), Arg Arg Trp Trp Cys Ser (SEQ ID NO;

38), Arg Arg Trp Trp Cys Thr (SEQ ID NO;

39), Arg Arg Trp Trp Cys Val (SEQ ID NO;

40), Arg Arg Trp Trp Cys Trp (SEQ ID NO;

41) or Arg Arg Trp Trp Cys Tyr (SEQ ID NO;

42).

7. The method of claim 5, wherein the at least one peptide comprises the amino acid sequence Arg Arg Trp Trp Cys Arg (SEQ ID NO:

1), Arg Arg Trp Trp Cys Ala (SEQ ID NO;

24), Arg Arg Trp Trp Cys Cys (SEQ ID NO;

25), Arg Arg Trp Trp Cys Phe (SEQ ID NO;

28), Arg Arg Trp Trp Cys Gly (SEQ ID NO;

29), Arg Arg Trp Trp Cys His (SEQ ID NO;

30), Arg Arg Trp Trp Cys Ile (SEQ ID NO;

31), Arg Arg Trp Trp Cys Lys (SEQ ID NO;

32), Arg Arg Trp Trp Cys Leu (SEQ ID NO;

33), Arg Arg Trp Trp Cys Met (SEQ ID NO;

34), Arg Arg Trp Trp Cys Pro (SEQ ID NO;

36), Arg Arg Trp Trp Cys Ser (SEQ ID NO;

38), Arg Arg Trp Trp Cys Thr (SEQ ID NO;

39), Arg Arg Trp Trp Cys Val (SEQ ID NO;

40), Arg Arg Trp Trp Cys Trp (SEQ ID NO;

41) or Arg Arg Trp Trp Cys Tyr (SEQ ID NO;

42).

8. The method of claim 5, wherein the at least one peptide comprises the amino acid sequence Arg Arg Trp Trp Cys Arg (SEQ ID NO:

1).

9. The method of claim 5, wherein the at least one peptide comprises the amino acid sequence Arg Arg Trp Trp Cys Lys (SEQ ID NO:

32).

10. The method of claim 8, wherein the at least one peptide comprises the amino acid sequence Arg Arg Trp Trp Cys Arg Xaa₂ (SEQ ID NO:

2), wherein Xaa₂ is any amino acid residue.

11. The method of claim 10, wherein the at least one peptide comprises the amino acid sequence Arg Arg Trp Trp Cys Arg Ala (SEQ ID NO:

3);

Arg Arg Trp Trp Cys Arg Cys (SEQ ID NO;

4);

Arg Arg Trp Trp Cys Arg Asp (SEQ ID NO;

5);

Arg Arg Trp Trp Cys Arg Glu (SEQ ID NO;

6);

Arg Arg Trp Trp Cys Arg Phe (SEQ ID NO;

7);

Arg Arg Trp Trp Cys Arg Gly (SEQ ID NO;

8);

Arg Arg Trp Trp Cys Arg His (SEQ ID NO;

9);

Arg Arg Trp Trp Cys Arg Ile (SEQ ID NO;

10);

Arg Arg Trp Trp Cys Arg Lys (SEQ ID NO;

11);

Arg Arg Trp Trp Cys Arg Leu (SEQ ID NO;

12);

Arg Arg Trp Trp Cys Arg Met (SEQ ID NO;

13);

Arg Arg Trp Trp Cys Arg Asn (SEQ ID NO;

14);

Arg Arg Trp Trp Cys Arg Pro (SEQ ID NO;

15);

Arg Arg Trp Trp Cys Arg Gln (SEQ ID NO;

16);

Arg Arg Trp Trp Cys Arg Arg (SEQ ID NO;

17);

Arg Arg Trp Trp Cys Arg Ser (SEQ ID NO;

18);

Arg Arg Trp Trp Cys Arg Thr (SEQ ID NO;

19);

Arg Arg Trp Trp Cys Arg Val (SEQ ID NO;

20), Arg Arg Trp Trp Cys Arg Trp (SEQ ID NO;

21) or Arg Arg Trp Trp Cys Arg Tyr (SEQ ID NO;

22).

12. The method of claim 11, wherein the at least one peptide comprises the amino acid sequence Arg Arg Trp Trp Cys Arg Cys (SEQ ID NO:

4);

Arg Arg Trp Trp Cys Arg Asp (SEQ ID NO;

5);

Arg Arg Trp Trp Cys Arg Phe (SEQ ID NO;

7);

Arg Arg Trp Trp Cys Arg Gly (SEQ ID NO;

8);

Arg Arg Trp Trp Cys Arg His (SEQ ID NO;

9);

Arg Arg Trp Trp Cys Arg Lys (SEQ ID NO;

11);

Arg Arg Trp Trp Cys Arg Asn (SEQ ID NO;

14);

Arg Arg Trp Trp Cys Arg Arg (SEQ ID NO;

17);

Arg Arg Trp Trp Cys Arg Trp (SEQ ID NO;

21).

13. The method of claim 1, wherein the composition comprises a biologically functional equivalent of said at least one peptide.

14. The method of claim 13, wherein said biologically functional equivalent comprises the amino acid sequence Gln Ile Pro Arg Arg Ser Trp Cys Arg Phe Leu Phe (SEQ ID NO:

52).

15. The method of claim 1, wherein the at least one peptide is an acylated peptide.

16. The method of claim 1, wherein the at least one peptide is acetylated at the C-terminus.

17. The method of claim 1, wherein the at least one peptide is acetylated at the N-terminus and amidated at the C-terminus.

18. The method of claim 1, wherein the at least one peptide comprises at least one D-amino acid.

19. The method of claim 1, wherein the at least one peptide is from six to about 15 residues in length.

20. The method of claim 1, wherein the at least one peptide comprises more than one peptide.

21. The method of claim 20, wherein said more than one peptide comprises up to 45 distinct peptides.

22. The method of claim 1, wherein the composition is a pharmaceutical composition in a pharmacologically acceptable form.

23. The method of claim 22, wherein said pharmaceutical composition further comprises an additional desired ingredient.

24. The method of claim 23, wherein the additional desired ingredient is IL-8 derived N-terminal peptides, IFN-γ

or oxygen radical scavengers.

25. The method of claim 1, wherein said target cells are comprised in a patient.

26. The method of claim 1, wherein said patient is a human.

27. The method of claim 1, wherein said method inhibits inflammation.

28. The method of claim 1, wherein said target cells comprise granulocyte cells or neutrophil cells.

29. The method of claim 1, wherein said method inhibits tumor cell proliferation.

30. The method of claim 1, wherein said target cells comprise at least one tumor cell.

31. The method of claim 30, wherein at least one tumor cell is a melanoma cell, prostate cancer cell or lung cancer cell.

32. A method of inhibiting inflammation, comprising administering to a patient, in an amount effective to inhibit at least one type of inflammation, a pharmaceutical composition comprising at least one peptide that comprises the amino acid sequence Arg Arg X Trp Cys X (SEQ ID NO:

58), or a biologically functional equivalents thereof, wherein X is any amino acid.

33. The method of claim 32, wherein said inflammation comprises lung inflammation, a skin disorder, a disease of the joint, pseudogout, inflammatory bowel disease or reperfusion cardiac damage.

34. The method of claim 33, wherein said lung injury or disorder comprises lung inflammation.

35. The method of claim 34, wherein said lung inflammation comprises asthma, bronchitis, cystic fibrosis or adult respiratory distress syndrome.

36. The method of claim 32, wherein said at least one peptide comprises the amino acid sequence Arg Arg Trp Trp Cys Xaa₁ (SEQ ID NO:

23), wherein Xaa₁ is any amino acid.

37. The method of claim 36, wherein the at least one peptide comprises the amino acid sequence Arg Arg Trp Trp Cys Arg (SEQ ID NO:

1), Arg Arg Trp Trp Cys Ala (SEQ ID NO;

24), Arg Arg Trp Trp Cys Cys (SEQ ID NO;

25), Arg Arg Trp Trp Cys Asp (SEQ ID NO;

26), Arg Arg Trp Trp Cys Glu (SEQ ID NO;

27), Arg Arg Trp Trp Cys Phe (SEQ ID NO;

28), Arg Arg Trp Trp Cys Gly (SEQ ID NO;

29), Arg Arg Trp Trp Cys His (SEQ ID NO;

30), Arg Arg Trp Trp Cys Ile (SEQ ID NO;

31), Arg Arg Trp Trp Cys Lys (SEQ ID NO;

32), Arg Arg Trp Trp Cys Leu (SEQ ID NO;

33), Arg Arg Trp Trp Cys Met (SEQ ID NO;

34), Arg Arg Trp Trp Cys Asn (SEQ ID NO;

35), Arg Arg Trp Trp Cys Pro (SEQ ID NO;

36), Arg Arg Trp Trp Cys Gln (SEQ ID NO;

37), Arg Arg Trp Trp Cys Ser (SEQ ID NO;

38), Arg Arg Trp Trp Cys Thr (SEQ ID NO;

39), Arg Arg Trp Trp Cys Val (SEQ ID NO;

40), Arg Arg Trp Trp Cys Trp (SEQ ID NO;

41) or Arg Arg Trp Trp Cys Tyr (SEQ ID NO;

42).

38. The method of claim 37, wherein the peptide comprises the amino acid sequence Arg Arg Trp Trp Cys Arg Xaa₂ (SEQ ID NO:

2), wherein Xaa₂ is any amino acid residue.

39. The method of claim 38, wherein the peptide comprises the amino acid sequence Arg Arg Trp Trp Cys Arg Ala (SEQ ID NO:

3);

Arg Arg Trp Trp Cys Arg Cys (SEQ ID NO;

4);

Arg Arg Trp Trp Cys Arg Asp (SEQ ID NO;

5);

Arg Arg Trp Trp Cys Arg Glu (SEQ ID NO;

6);

Arg Arg Trp Trp Cys Arg Phe (SEQ ID NO;

7);

Arg Arg Trp Trp Cys Arg Gly (SEQ ID NO;

8);

Arg Arg Trp Trp Cys Arg His (SEQ ID NO;

9);

Arg Arg Trp Trp Cys Arg Ile (SEQ ID NO;

10);

Arg Arg Trp Trp Cys Arg Lys (SEQ ID NO;

1);

Arg Arg Trp Trp Cys Arg Leu (SEQ ID NO;

12);

Arg Arg Trp Trp Cys Arg Met (SEQ ID NO;

13);

Arg Arg Trp Trp Cys Arg Asn (SEQ ID NO;

14);

Arg Arg Trp Trp Cys Arg Pro (SEQ ID NO;

15);

Arg Arg Trp Trp Cys Arg Gln (SEQ ID NO;

16);

Arg Arg Trp Trp Cys Arg Arg (SEQ ID NO;

17);

Arg Arg Trp Trp Cys Arg Ser (SEQ ID NO;

18);

Arg Arg Trp Trp Cys Arg Thr (SEQ ID NO;

19);

Arg Arg Trp Trp Cys Arg Val (SEQ ID NO;

20);

Arg Arg Trp Trp Cys Arg Trp (SEQ ID NO;

21) or Arg Arg Trp Trp Cys Arg Tyr (SEQ ID NO;

22).

40. A method of inhibiting tumor cell proliferation, comprising administering to a patient, in an amount effective to inhibit at least one type of tumor cell proliferation, a pharmaceutical composition comprising at least one peptide that comprises the amino acid sequence Arg Arg X Trp Cys X (SEQ ID NO:

58), or a biologically functional equivalents thereof, wherein X is any amino acid.

41. The method of claim 40, wherein said tumor cell is a melanoma cell, prostate cancer cell or lung cancer cell.

42. The method of claim 40, wherein said at least one peptide comprises the amino acid sequence Arg Arg Trp Trp Cys Xaa₁ (SEQ ID NO:

23), wherein Xaa₁ is any amino acid.

43. The method of claim 42, wherein the at least one peptide comprises the amino acid sequence Arg Arg Trp Trp Cys Arg (SEQ ID NO:

1), Arg Arg Trp Trp Cys Ala (SEQ ID NO;

24), Arg Arg Trp Trp Cys Cys (SEQ ID NO;

25), Arg Arg Trp Trp Cys Asp (SEQ ID NO;

26), Arg Arg Trp Trp Cys Glu (SEQ ID NO;

27), Arg Arg Trp Trp Cys Phe (SEQ ID NO;

28), Arg Arg Trp Trp Cys Gly (SEQ ID NO;

29), Arg Arg Trp Trp Cys His (SEQ ID NO;

30), Arg Arg Trp Trp Cys Ile (SEQ ID NO;

31), Arg Arg Trp Trp Cys Lys (SEQ ID NO;

32), Arg Arg Trp Trp Cys Leu (SEQ ID NO;

33), Arg Arg Trp Trp Cys Met (SEQ ID NO;

34), Arg Arg Trp Trp Cys Asn (SEQ ID NO;

35), Arg Arg Trp Trp Cys Pro (SEQ ID NO;

36), Arg Arg Trp Trp Cys Gln (SEQ ID NO;

37), Arg Arg Trp Trp Cys Ser (SEQ ID NO;

38), Arg Arg Trp Trp Cys Thr (SEQ ID NO;

39), Arg Arg Trp Trp Cys Val (SEQ ID NO;

40), Arg Arg Trp Trp Cys Trp (SEQ ID NO;

41) or Arg Arg Trp Trp Cys Tyr (SEQ ID NO;

42).

44. The method of claim 43, wherein the at least one peptide comprises the amino acid sequence Arg Arg Trp Trp Cys Arg Xaa₂ (SEQ ID NO:

2), wherein Xaa₂ is any amino acid residue.

45. The method of claim 44, wherein the at least one peptide comprises the amino acid sequence Arg Arg Trp Trp Cys Arg Ala (SEQ ID NO:

3);

Arg Arg Trp Trp Cys Arg Cys (SEQ ID NO;

4);

Arg Arg Trp Trp Cys Arg Asp (SEQ ID NO;

5) Arg Arg Trp Trp Cys Arg Glu (SEQ ID NO;

6);

Arg Arg Trp Trp Cys Arg Phe (SEQ ID NO;

7);

Arg Arg Trp Trp Cys Arg Gly (SEQ ID NO;

8);

Arg Arg Trp Trp Cys Arg His (SEQ ID NO;

9);

Arg Arg Trp Trp Cys Arg Ile (SEQ ID NO;

10);

Arg Arg Trp Trp Cys Arg Lys (SEQ ID NO;

11);

Arg Arg Trp Trp Cys Arg Leu (SEQ ID NO;

12);

Arg Arg Trp Trp Cys Arg Met (SEQ ID NO;

13);

Arg Arg Trp Trp Cys Arg Asn (SEQ ID NO;

14);

Arg Arg Trp Trp Cys Arg Pro (SEQ ID NO;

15);

Arg Arg Trp Trp Cys Arg Gln (SEQ ID NO;

16);

Arg Arg Trp Trp Cys Arg Arg (SEQ ID NO;

17);

Arg Arg Trp Trp Cys Arg Ser (SEQ ID NO;

18);

Arg Arg Trp Trp Cys Arg Thr (SEQ ID NO;

19);

Arg Arg Trp Trp Cys Arg Val (SEQ ID NO;

20);

Arg Arg Trp Trp Cys Arg Trp (SEQ ID NO;

21) or Arg Arg Trp Trp Cys Arg Tyr (SEQ ID NO;

22).