Specific inhibitors of NFAT activation by calcineurin and their use in treating immune-related diseases

• US 7,816,490 B2
• Filed: 10/28/2008
• Issued: 10/19/2010
• Est. Priority Date: 02/12/1998
• Status: Expired due to Term

• Current Assignee: Immune Disease Institute, Inc.
• Original Assignee: Immune Disease Institute, Inc.
• Inventors: Hogan, Patrick G., Aramburu, Jose, Rao, Anjana
• Primary Examiner(s): Nickol; Gary B.
• Assistant Examiner(s): Stoica; Elly-Gerald
• Application Number:
  US12/260,049
• Publication Number:
  US 20090186422A1
• Time in Patent Office: 721 Days
• Field of Search: None
• US Class Current: 530/330
• 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: C07K 5/101 the side chain containing 2 to 4 carbon atoms, e.g. Val, Ile, Leu

Abstract

Isolated peptide fragments of the conserved regulatory domain of NFAT protein capable of inhibiting protein-protein interaction between calcineurin and NFAT, or a biologically active analog thereof are described. Isolated polynucleotides and gene therapy vectors encoding such peptide fragments are also described. In addition, methods for treating immune-related diseases or conditions and methods for high throughput screening of candidate agents are described. Pharmaceutical compositions are also provided.

Claims

1. An NFAT derivative comprising the sequence of SEQ ID NO:

22, wherein at least one non-conserved residue in SEQ ID NO;

22 is substituted, and wherein the NFAT derivative is less than 50 amino acid residues long and binds to calcineurin.

2. The NFAT derivative of claim 1, wherein the NFAT derivative is less than 20 amino acid residues long.

13. The NFAT derivative of claim 1, wherein the NFAT derivative is 13 amino acid residues long.

3. A peptide inhibitor of protein-protein interaction between calcineurin and NFAT identified by a process comprising the steps of:

providing a first compound comprising NFAT or a biologically active derivative thereof;

providing a second compound comprising calcineurin or a biologically active derivative thereof;

providing a candidate agent;

contacting said first compound, said second compound and said candidate agent with each other;

determining the amount of said second compound bound to said first compound or determining the amount of said first compound bound to said second compound, wherein a reduction in binding between said first compound and said second compound indicates that said candidate agent is an inhibitor of protein-protein interaction between calcineurin and NFAT; and

wherein the peptide inhibitor is an NFAT derivative comprising the sequence of SEQ ID NO;

22, wherein at least one non-conserved residue in SEQ ID NO;

22 is substituted, and wherein the NFAT derivative is less than 150 amino acid residues long and binds to calcineurin.

4. The peptide inhibitor of claim 3, wherein the NFAT derivative is less than 20 amino acid residues long.

14. The peptide inhibitor of claim 3, wherein the peptide inhibitor is less than 100 amino acid residues long.

15. The peptide inhibitor of claim 3, wherein the peptide inhibitor is less than 50 amino acid residues long.

16. The peptide inhibitor of claim 3, wherein the peptide inhibitor is 13 amino acid residues long.

5. An NFAT derivative comprising the sequence of SEQ ID NO:

5, wherein at least one non-conserved residue in SEQ ID NO;

5 is substituted, and wherein the NFAT derivative is less than 50 amino acid residues long and binds to calcineurin.

6. The NFAT derivative of claim 5, wherein the NFAT derivative is less than 20 amino acid residues long.

7. The NFAT derivative of claim 5, wherein the NFAT derivative is less than 10 amino acid residues long.

8. The NFAT derivative of claim 5, wherein the NFAT derivative is 6 amino acid residues long.

9. A peptide inhibitor of protein-protein interaction between calcineurin and NFAT identified by a process comprising the steps of:

providing a first compound comprising NFAT or a biologically active derivative thereof;

providing a second compound comprising calcineurin or a biologically active derivative thereof;

providing a candidate agent;

contacting said first compound, said second compound and said candidate agent with each other;

determining the amount of said second compound bound to said first compound or determining the amount of said first compound bound to said second compound, wherein a reduction in binding between said first compound and said second compound indicates that said candidate agent is an inhibitor of protein-protein interaction between calcineurin and NFAT; and

wherein the peptide inhibitor is an NFAT derivative comprising the sequence of SEQ ID NO;

5, wherein at least one non-conserved residue in SEQ ID NO;

5 is substituted, and wherein the NFAT derivative is less than 150 amino acid residues long and binds to calcineurin.

10. The peptide inhibitor of claim 9, wherein the NFAT derivative is less than 20 amino acid residues long.

11. The peptide inhibitor of claim 9, wherein the peptide inhibitor is less than 10 amino acid residues long.

12. The peptide inhibitor of claim 9, wherein the peptide inhibitor is 6 amino acid residues long.

17. The peptide inhibitor of claim 9, wherein the NFAT derivative is less than 100 amino acid residues long.

18. The peptide inhibitor of claim 9, wherein the NFAT derivative is less than 50 amino acid residues long.