High affinity mutants of nuclear factor-interleukin 6 and methods of use therefor

• US 5,804,445 A
• Filed: 01/11/1996
• Issued: 09/08/1998
• Est. Priority Date: 01/11/1996
• 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: Brasier, Allan R.
• Primary Examiner(s): Robinson, Douglas W.
• Assistant Examiner(s): Nelson, Amy J.
• Application Number:
  US08/585,197
• Time in Patent Office: 971 Days
• Field of Search: 530/324, 514/2, 514/44, 435/375, 435/325, 435/243, 424/93.2
• US Class Current: 435/375
• 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 14/4705 stimulating, promoting or activating activity

Abstract

The present invention relates to inhibitors of the sequence specific transcription factor nuclear factor IL-6 (NF-IL6) and methods of use therefor. In particular, substitution mutants in the N-terminus of the NF-IL6 tryptic core domain are disclosed that have a higher binding affinity for the DNA binding site than does the wild-type sequence.

Claims

1. An isolated polypeptide comprising an NF-IL6 tryptic core domain, wherein the N-terminus of said core domain has a net charge that is less negative than the N-terminus of wild-type nuclear factor interleukin 6 (NF-IL6), and wherein the binding affinity of said polypeptide is higher than NF-IL6 for its target sequence.

2. The isolated polypeptide according to claim 1, wherein said tryptic core domain corresponds to residues 266 to 345 of wild-type NF-IL6 tryptic core domain.

3. The isolated polypeptide according to claim 1, wherein said N-terminus corresponds to residues 266 to 272 of wild-type NF-IL6 tryptic core domain.

4. The isolated polypeptide according to claim 3, wherein said N-terminus is at least one unit charge less negatively charged than wild-type NF-IL6 tryptic core domain.

5. The isolated polypeptide according to claim 3, wherein the N-terminus lacks one of the aspartic acid residues found in the wild-type NF-IL6 tryptic core domain and has substituted therefore an uncharged or positively charged residue.

6. The isolated polypeptide according to claim 3, wherein the N-terminus lacks both aspartic acid residues found in the wild-type NF-IL6 tryptic core domain and has substituted therefore at least one uncharged or positively charged residue.

7. The isolated polypeptide according to claim 5, wherein the substitution is an alanine, glycine or threonine residue.

8. The isolated polypeptide according to claim 5, wherein the substitution is an alanine residue.

9. The polypeptide according to claim 8, wherein the substitution is alanine for aspartic acid at position 268.

10. The polypeptide according to claim 8, wherein the substitution is alanine for aspartic acid at position 272.

11. The isolated polypeptide according to claim 6, wherein the substitution is alanine, glycine or threonine residues.

12. The isolated polypeptide according to claim 6, wherein the substitution is alanine residues.

13. The isolated polypeptide according to claim 6, wherein both aspartic acid residues are substituted with uncharged or positively charged residues.

14. The isolated polypeptide according to claim 13, wherein the substitutions are alanine, glycine or threonine residues.

15. The isolated polypeptide according to claim 13, wherein the substitutions are alanine residues.

16. A composition comprising:

(i) a polypeptide comprising an NF-IL6 tryptic core domain, wherein the N-terminus of said core domain has a net charge that is less negative that wild-type NF-IL6 tryptic core domain, and wherein the binding affinity of said polypeptide is higher than NF-IL6 for its target sequence; and

(ii) a pharmaceutically acceptable carrier.

17. The composition according to claim 16, wherein said polypeptide is formulated with a liposome.

18. The composition according to claim 16, wherein said polypeptide is formulated with folate-conjugated bovine serum albumin.

19. A method for inhibiting NF-IL6 function in a cell in culture comprising the steps of:

(i) providing a polypeptide comprising an NF-IL6 tryptic core domain, wherein the N-terminus of said core domain has a net charge that is less negative than wild-type NF-IL6 tryptic core domain, and wherein the binding affinity of said polypeptide is higher than NF-IL6 for its target sequence; and

(ii) contacting said cell with said polypeptide.

20. The method according to claim 19, wherein said polypeptide is formulated with a liposome.

21. The method according to claim 19, wherein said polypeptide is formulated with folate-conjugated bovine serum albumin.