BH3 peptides and method of use thereof

• US 20080234201A1
• Filed: 04/24/2007
• Published: 09/25/2008
• Est. Priority Date: 09/09/2002
• Status: Active Grant

• Current Assignee: Dana-Farber Cancer Institute
• Original Assignee: Dana-Farber Cancer Institute
• Inventors: Letai, Anthony, Korsmeyer, Stanley J.
• Granted Patent: US 7,868,133 B2
• Time in Patent Office: Days
• US Class Current: 514/13
• 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: C07K 14/4747 Apoptosis related proteins
• CPC Class Codes: C07K 2319/03 containing a transmembrane segment

Abstract

The invention provides peptides and the nucleic acid sequences that encode them. The invention further provides therapeutic, diagnostic and research methods for diagnosis, treatment, and prevention of apoptosis associated disorders.

Claims

1. An isolated peptide comprising the amino acid sequence selected from the group consisting of SEQ ID NO:

1, 2 or 10.

2. The peptide of claim 1, wherein said peptide induces BAK oligomerization and cytochrome c release from mitochondria.

8. A nucleic acid encoding the peptide of any one of claims 1-7.

9. An expression vector comprising the nucleic acid of claim 8 operably linked to a promoter.

10. A host cell containing the expression vector of claim 9.

11. A composition comprising a peptide of any one of claims 1-7 and a carrier.

12. A method of treating a cell proliferative disorder in a subject comprising administering to a subject in need thereof a composition comprising the peptide of any one of claims 1-7.

13. The method of claim 12, wherein in said cell proliferative disorder is cancer.

14. The method of claim 12, wherein the composition is further administered with a chemotherapeutic compound.

3. An isolated peptide comprising the amino acid sequence selected from the group consisting of SEQ ID NOs:

3-7 or 11.

4. The peptide of claim 3, wherein said peptide binds BCL-2 or MCL-2.

5. An isolated peptide of any one of SEQ ID NOs:

1-11.

6. A chimeric peptide comprising a first domain and a second domain wherein said first domain comprises the amino acid sequence selected from the group consisting of and SEQ ID NOs:

1-11 and said second domain comprising a translocation sequence which facilitates transport across a biological membrane.

7. The peptide of claim 6, wherein said translocation sequence is polyarginine.

15. A method of inducing apoptosis in a cell comprising contacting said cell with a composition comprising any if of SEQ ID NOs 1, 2 or 10 in an amount sufficient to induce apoptosis in said cell.

16. A method of sensitizing a cell to apoptosis comprising contacting said cell with a composition comprising any if of SEQ ID NOs:

2-7 or 11 in an amount sufficient to sensitize said cell to apoptosis.

17. A method of screening for an apoptotic sensitizer compound comprising:

(a) contacting mitochondria overexpressing an anti-apoptotic protein with a BID-like BH3 peptide to form a protein-peptide complex;

(b) contacting said complex with a test compound; and

(c) determining cytochrome c release from said mitochondria, wherein an increase of cytochrome c release in the presence of said test compound compared to the absence of said compound indicates said compound is an apoptotic sensitizer compound

18. The method of claim 17, wherein said BID-like BH3 peptide is wildtype BID or a fragment thereof.

19. The method of claim 17, wherein said anti-apoptotic protein is BCL-2.

20. A transgenic non-human animal comprising a recombinant BCL-2 nucleic acid molecule stably integrated into the genome of said animal.

21. The animal of claim 20, wherein said recombinant nucleic acid molecule is operably linked to one or more regulatory sequences.

23. The animal of claim 20, wherein said recombinant nucleic acid molecule is of human or murine origin.

24. An isolated cell of the animal of claim 20.

25. The cell of claim 24, wherein said cell is a stem cell, a germ cell, a precursor cell or a progenitor cell.

26. The animal of claim 20, wherein said animal is a rodent.

31. A method for the identification of a compound capable of modifying an activity of a BCL-2 protein, comprising:

(a) contacting the transgenic non-human animal of claim 20 or a cell therefrom with a test compound; and

(b) measuring the effect of said test compound on said BCL-2 protein;

thereby identifying a compound that modifies an activity of said protein.

32. The method of claim 31, wherein said test compound is a BH-3 agonist.

22. The animal of claim 22, wherein said further regulatory sequence is a promoter.

27. The animal of claim 27, wherein said rodent is a mouse.

28. A method for the production of a transgenic non-human animal, comprising introduction of a recombinant BCL-2 nucleic acid molecule into a germ cell, an embryonic cell, an egg cell or a cell derived therefrom.

29. The method of claim 28, wherein said animal is a rodent.

30. The method of claim 29, wherein said rodent is a mouse.