14-3-3 Binding molecules as sensitizers for anticancer therapies

• US 20040152630A1
• Filed: 03/19/2004
• Published: 08/05/2004
• Est. Priority Date: 02/21/2001
• Status: Active Grant

• Current Assignee: Emory University
• Original Assignee: Emory University
• Inventors: Masters, Shane C., Fu, Haian
• Granted Patent: US 7,335,635 B2
• Time in Patent Office: Days
• US Class Current: 514/12
• CPC Class Codes: A61K 2300/00 Mixtures or combinations of active ingredients, wherein at least one active ingredient is fully defined in groups A61K31/00 - A61K41/00
• CPC Class Codes: A61K 31/704 attached to a condensed carbocyclic ring system, e.g. sennosides, thiocolchicosides, escin, daunorubicin digitoxin A61K31/7048
• CPC Class Codes: A61K 31/7048 having oxygen as a ring hetero atom, e.g. leucoglucosan, hesperidin, erythromycin, nystatin , digitoxin or digoxin
• CPC Class Codes: A61K 31/7072 having two oxo groups directly attached to the pyrimidine ring, e.g. uridine, uridylic acid, thymidine, zidovudine
• CPC Class Codes: A61K 38/10 Peptides having 12 to 20 amino acids A61K38/043 - A61K38/046 take precedence
• CPC Class Codes: A61K 38/16 Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof enzyme inhibitors A61K38/005
• CPC Class Codes: A61K 38/17 from animals; from humans enzyme inhibitors A61K38/005
• CPC Class Codes: A61K 45/06 Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: C07K 14/00 Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
• CPC Class Codes: G01N 2500/00 Screening for compounds of potential therapeutic value

Abstract

Methods are provided for enhancing the death of a neaplastic cell comprising the administration of a therapeutically effective concentration of a 14-3-3 antagonist and at least one antineoplastic therapeutic agent. The methods of the invention find use in improving the clinical outcome of a mammal having a neoplastic disorder and comprises administration to a mammal in need thereof at least one antineoplastic therapeutic agent in combination with a 14-3-3 antagonist. Further provided are pharmaceutical compositions having a therapeutically effective amount of a 14-3-3 antagonist and an antineoplastic therapeutic agent. Also provided are methods for identifying agents that selectively inhibit an interaction between a 14-3-3 polypeptide and a 14-3-3 ligand.

Claims

1. A method of enhancing the death of a neoplastic cell comprising providing to said neoplastic cell a therapeutically effective amount of a 14-3-3 antagonist and at least one antineoplastic therapeutic agent.

2. The method of claim 1, wherein said 14-3-3 antagonist disrupts an interaction between a 14-3-3 polypeptide and a 14-3-3 ligand.

3. The method of claim 2, wherein the 14-3-3 antagonist directly interacts with a 14-3-3 polypeptide.

4. The method of claim 3, wherein the 14-3-3 antagonist is a 14-3-3 amphipathic groove binding antagonist.

5. The method of claim 1, wherein said antineoplastic therapeutic agent is a chemotherapeutic agent.

6. The method of claim 5, wherein said chemotherapeutic agent is selected from the group consisting of etoposide, cisplatin, doxorubicin, paclitaxel, methotrexate, and 5-fluorouracil.

7. The method of claim 1, wherein said antineoplastic therapeutic agent is a radiation.

8. The method of claim 1, wherein said 14-3-3 antagonist comprises an amino acid sequence set forth in SEQ ID NO:

1, 2, or a biologically active variant thereof.

9. The method of claim 8, wherein said 14-3-3 antagonist comprises an amino acid sequence selected from the group consisting of:

a) an amino acid sequence having at least 80% sequence identity to the sequence of SEQ ID NO;

1 or 2, wherein said sequence disrupts the interaction between a 14-3-3 polypeptide and a 14-3-3 ligand; and

,

b) an amino acid sequence having at least 10 contiguous amino acids of SEQ ID NO;

1 or 2, wherein said sequence disrupts the interaction between a 14-3-3 polypeptide and a 14-3-3 ligand.

10. The method of claim 1, wherein the 14-3-3 antagonist and the antineoplastic therapeutic agent are provided to the cell sequentially.

11. The method of claim 1, wherein the 14-3-3 antagonist and the antineoplastic therapeutic agent are provided to the cell simultaneously.

12. The method of claim 1, wherein said neoplastic cell is in a mammal.

13. The method of claim 13, wherein said mammal is a human.

14. A method of treating a neoplastic disorder comprising administering to a mammal in need thereof a therapeutically effective amount of a 14-3-3 antagonist and at least one antineoplastic therapeutic agent.

15. A pharmaceutical composition comprising a therapeutically effective amount of a 14-3-3 antagonist and at least one antineoplastic therapeutic agent.

16. The pharmaceutical composition of claim 15, wherein said antineoplastic therapeutic agent is a chemotherapeutic agent, a biological response modifier, or a radiation.

17. The pharmaceutical composition of claim 16, wherein said chemotherapeutic agent is selected from the group consisting of etoposide, cisplatin, doxorubicin, paclitaxel, methotrexate, and 5-fluorouracil.

18. The pharmaceutical composition of claim 15, wherein said 14-3-3 antagonist disrupts an interaction between a 14-3-3 polypeptide and a 14-3-3 ligand.

19. The pharmaceutical composition of claim 18, wherein the 14-3-3 antagonist directly interacts with a 14-3-3 polypeptide.

20. The pharmaceutical composition of claim 19, wherein the 14-3-3 antagonist is a 14-3-3 amphipathic groove binding antagonist.

21. The pharmaceutical composition of claim 20, wherein said 14-3-3 antagonist comprises an isolated polypeptide comprising an amino acid sequence of SEQ ID NO:

1, 2, or a biologically active variant thereof.

22. The pharmaceutical composition of claim 21, wherein said polypeptide comprises an amino acid sequence selected from the group consisting of:

a) an amino acid sequence having at least 80% sequence identity to the sequence of SEQ ID NO;

1 or 2, wherein said sequence disrupts an interaction between a 14-3-3 polypeptide and a 14-3-3 ligand; and

,

b) an amino acid sequence having at least 10 contiguous amino acids of SEQ ID NO;

1 or 2, wherein said sequence disrupts an interaction between a 14-3-3 polypeptide and a 14-3-3 ligand.

23. A method of identifying an agent that selectively inhibits an interaction between a 14-3-3 polypeptide and a 14-3-3 ligand comprising:

a) contacting a 14-3-3 polypeptide with a 14-3-3 antagonist under conditions that permit formation of a 14-3-3/antagonist complex, said 14-3-3 antagonist comprises a polypeptide having an amino acid sequence selected from the group consisting of;

i) an amino acid sequence set forth in SEQ ID NO;

2 or a biologically active variant thereof;

ii) an amino acid sequence having at least 80% sequence identity to the sequence of SEQ ID NO;

2, wherein said sequence disrupts an interaction between a 14-3-3 polypeptide and a 14-3-3 ligand; and

,

iii) an amino acid sequence having at least 10 contiguous amino acids of SEQ ID NO;

2, wherein said sequence disrupts an interaction between a 14-3-3 polypeptide and a 14-3-3 ligand;

b) contacting said 14-3-3/antagonist complex with a candidate agent; and

,

c) determining if the candidate agent disrupts the 14-3-3/antagonist complex.

25. The method of claim 23 and 24, wherein said method occurs in vivo or in vitro.

24. A method of identifying an agent that selectively inhibits an interaction between a 14-3-3 polypeptide and a 14-3-3 ligand comprising:

a) contacting a 14-3-3 polypeptide with a candidate agent under conditions that allow for a 14-3-3/candidate agent complex to form;

b) contacting said 14-3-3/candidate agent complex with a 14-3-3 antagonist, said 14-3-3 antagonist comprises a polypeptide having an amino acid sequence selected from the group consisting of;

i) an amino acid sequence set forth in SEQ ID NO;

1, 2, or a biologically active variant thereof;

ii) an amino acid sequence having at least 80% sequence identity to the sequence of SEQ ID NO;

1 or 2, wherein said sequence disrupts an interaction between a 14-3-3 polypeptide and a 14-3-3 ligand; and

,

iii) an amino acid sequence having at least 10 contiguous amino acids of SEQ ID NO;

1 or 2, wherein said sequence disrupts an interaction between a 14-3-3 polypeptide and a 14-3-3 ligand; and

,

c) determining if the 14-3-3 antagonist disrupts the 14-3-3/candidate agent complex.

26. An isolated polypeptide comprising an amino acid sequence selected from the group consisting of:

a) an amino acid sequence set forth in SEQ ID NO;

2;

b) an amino acid sequence having at least 80% sequence identity to the sequence of SEQ ID NO;

2, wherein the sequence disrupts the interaction of a 14-3-3 ligand with a 14-3-3 polypeptide; and

,

c) an amino acid sequence having at least 19 contiguous amino acids of SEQ ID NO;

2, wherein the sequence disrupts the interaction of a 14-3-3 ligand with a 14-3-3 polypeptide.