Recombinant soluble truncated IL-23 receptor (IL-23R) capable of inhibiting IL-23R-mediated cell signaling
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.
1. A pharmaceutical composition useful in inhibiting IL-23R mediated cell signaling, comprising a recombinant IL-23Rα protein, said recombinant IL-23Rα protein consists of the amino acid sequence of SEQ ID NO: 4, and a pharmaceutical acceptable excipient.
2. The pharmaceutical composition of claim 1 , wherein said pharmaceutical composition is a solid form selected from the group consisting of pills, capsules, granules, tablets and powders.
3. The pharmaceutical composition of claim 1 , wherein said pharmaceutical composition is a liquid form selected from the group consisting of solutions, syrups, elixirs and suspensions.
4. The pharmaceutical composition of claim 1 , wherein said pharmaceutical composition is a parenteral form.
5. The pharmaceutical composition of claim 1 , wherein said pharmaceutical composition is a nasal form.
6. The pharmaceutical composition of claim 1 , wherein said recombinant IL-23Rα is prepared by the steps of:
a) transfecting a cell with a vector containing a gene consisting of the nucleotide sequence of SEQ ID NO: 3;
b) allowing said transfected cell to produce said recombinant IL-23Rα protein; and
c) isolating said recombinant IL-23Rα protein.
7. The pharmaceutical composition of claim 6 , wherein said cell is selected from the group consisting of 293T, CHO and COS.
8. The pharmaceutical composition of claim 6 , wherein said cell is 293T cell.
9. The pharmaceutical composition of claim 6 , wherein said vector is selected from the group consisting of pcDNA 3.3.Topo, pcDNA 3.1, pCI, pSI, pTARhET, pPMR and pTK-Hyg.
10. The pharmaceutical composition of claim 6 , wherein said vector is pcDNA 3.3.
11. The pharmaceutical composition of claim 6 , wherein said isolating step c) is performed using ion-exchange column, reverse phase HPLC, chromatography on silica, cation-exchange resin, chromatofocusing, SDS-PAGE, ammonium sulfate, metal chelating column, or antibody column.
12. The pharmaceutical composition of claim 6 , wherein said isolating step c) is performed using an antibody column.
13. The pharmaceutical composition of claim 6 , wherein said isolating step c) is performed using an anti-FLAG column.
14. A method of preparing a pharmaceutical composition containing a recombinant IL-23Rα protein, said recombinant IL-23Rα protein consists of the amino acid sequence of SEQ ID NO: 4, comprising the steps of:
a) preparing a recombinant IL-23Rα protein consisting of the amino acid sequence of SEQ ID NO: 4, said preparing step is performed by the steps of:
(i) transfecting a cell with a vector containing a gene, said gene consists of the nucleic acid sequence of SEQ ID NO: 3;
(ii) allowing said transfected cell to produce said recombinant IL-23Rα protein; and
(iii) isolating said recombinant IL-23Rα protein; and
b) constituting said recombinant IL-23Rα protein with a pharmaceutically acceptable excipient to form a pharmaceutical composition.