Therapeutic application of isolated naturally-occuring soluble truncated forms of IL-23 receptor

• US 20120071422A1
• Filed: 03/31/2011
• Published: 03/22/2012
• Est. Priority Date: 03/31/2010
• Status: Active Grant

• Current Assignee: Medical Diagnostic Laboratories LLC
• Original Assignee: Medical Diagnostic Laboratories LLC
• Inventors: Gallagher, Grant, Yu, Raymond, Brazaitis, Jonathan
• Granted Patent: US 10,416,172 B2
• Time in Patent Office: Days
• US Class Current: 514/21.2
• 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 14/7155 for interleukins [IL]
• CPC Class Codes: C12N 5/0602 Vertebrate cells
• CPC Class Codes: G01N 2333/705 Assays involving receptors, cell surface antigens or cell surface determinants
• CPC Class Codes: G01N 2800/065 Bowel diseases, e.g. Crohn, ulcerative colitis, IBS
• CPC Class Codes: G01N 2800/368 Pregnancy complicated by disease or abnormalities of pregnancy, e.g. preeclampsia, preterm labour
• CPC Class Codes: G01N 33/6863 Cytokines, i.e. immune system proteins modifying a biological response such as cell growth proliferation or differentiation, e.g. TNF, CNF, GM-CSF, lymphotoxin, MIF or their receptors
• CPC Class Codes: G01N 33/6893 related to diseases not provided for elsewhere

Abstract

The present invention relates to an isolated naturally-occurring soluble truncated IL-23Rα protein, which is a translated protein resulting from a mRNA splice variant of IL-23Rα. The soluble IL-23Rα proteins (e.g., Δ9 and Δ8,9) represents a novel soluble IL-23Rα protein, which is lacking a transmembrane domain and has a unique eight (8) amino acids (GLKEGSYC) at its C-terminus end (due to frame-shift). ELISA reveals that Δ9 is present in blood and can serve as a diagnostic tool for auto-immune diseases including Crohn'"'"'s disease. There is also provided a method of recombinant production for this soluble truncated form of IL-23Rα protein. More importantly, the present invention provides an utility application of the Δ9 and Δ8,9 protein in inhibit IL-23R-mediated cell signaling. More particularly, Δ9 and Δ8,9 blocks STAT3 formation as well as Th17 maturation. There is provided a therapeutic application of Δ9 and Δ8,9 in treating a human patient inflicted with Crohn'"'"'s disease.

Claims

1. A recombinant IL-23Rα

protein, wherein said recombinant protein has an amino acid sequence selected from the group consisting of SEQ ID NO;

2 and SEQ ID NO;

4, and wherein said recombinant protein binds to IL-23.

2. The recombinant IL-23Rα

protein of claim 1, wherein said recombinant protein has an amino acid sequence of SEQ ID NO;

2.

3. The recombinant IL-23Rα

protein of claim 1, wherein said recombinant protein has an amino acid sequence of SEQ ID NO;

4.

4. The recombinant IL-23Rα

protein of claim 1, wherein said recombinant protein interferes with IL-23 cytokine binding to IL-23 receptor, and thereby inhibits IL-23R-mediated cell signaling.

5. The recombinant IL-23Rα

protein of claim 4, wherein said IL-23R-mediated cell signaling is the formation of at least one transcriptional factor selected from the group consisting of STAT1, STAT2, STAT3, and STAT5.

6. The recombinant IL-23Rα

protein of claim 5, wherein said IL-23R-mediated cell signaling is the STAT3 formation.

7. The recombinant IL-23Rα

protein of claim 4, wherein said IL-23R-mediated cell signaling is the formation of IL-17A or IL-17F.

8. The recombinant IL-23Rα

protein of claim 4, wherein said IL-23R-mediated cell signaling is the IL-17A formation.

9. The recombinant IL-23Rα

protein of claim 4, wherein said protein further has the ability to inhibit Th17 cell maturation.

10. A pharmaceutical composition comprising the recombinant protein of claim 1 and a pharmaceutical acceptable excipient.

11. A pharmaceutical composition comprising the recombinant protein of claim 2 and a pharmaceutical acceptable excipient.

12. A pharmaceutical composition comprising the recombinant protein of claim 3 and a pharmaceutical acceptable excipient.

13. A method of preparing a recombinant IL-23Rα

protein, comprising the steps of;

a) transfecting a cell with a gene encoding a protein that has an amino acid sequence selected from the group consisting of SEQ ID NO;

2 and SEQ ID NO;

4;

b) allowing said transfected cell to produce a recombinant IL-23Rα

protein; and

c) isolating said recombinant IL-23Rα

protein, wherein said recombinant IL-23Rα

protein has an amino acid sequence of SEQ ID NO;

2 or SEQ ID NO;

4.

14. The method of claim 13, wherein said recombinant IL-23Rα

protein has an amino acid sequence of SEQ ID NO;

2.

15. The method of claim 13, wherein said recombinant IL-23Rα

protein has an amino acid sequence of SEQ ID NO;

4.

16. The method of claim 13, wherein said gene has a nucleotide sequence selected from the group consisting of SEQ ID NO:

1 and SEQ ID NO;

3.

17. The method of claim 13, wherein said gene has a nucleotide sequence of SEQ ID NO:

1.

18. The method of claim 13, wherein said gene has a nucleotide sequence of SEQ ID NO:

3.

19. The method of claim 13, wherein said transfected cell is a mammalian cell.

20. The method of claim 13, wherein said recombinant IL-23Rα

protein is a soluble truncated IL-23Rα

protein, and has the following characteristics of;

a) lacking a transmembrane domain;

b) existing as a monomer; and

c) having the ability to inhibit IL-23R-mediated cell signaling.

21. A method of inhibiting IL-23R-mediated cell signaling in a mammalian cell, comprising the steps of:

a) exposing a mammalian cell to a recombinant IL-23Rα

protein, wherein said recombinant protein has an amino acid sequence selected from the group consisting of SEQ ID NO;

2 and SEQ ID NO;

4.

22. The method of claim 21, wherein said mammalian cell is a human cell.