Interleukin-9 antagonist muteins and their pharmacological methods of use
This invention relates to IL-9 muteins that inhibit the activity of wild-type IL-9, multimers and Fc-fusion constructs of IL-9 proteins, and an efficient method to purify IL-9 proteins produced by eukaryotic cells. Related formulations, dosages and methods of administration thereof for therapeutic purposes are also provided. More particularly, these IL-9 muteins, compositions, and methods provide a treatment option for individuals afflicted with conditions where inhibiting IL-9 mediated immune responses would be beneficial, such as allergy, asthma, chronic obstructive pulmonary disease (emphysema and chronic bronchitis), pulmonary and gastro-intestinal mucus hyperplasia, inflammation, immunological disorders, leukemia, and lymphoma.
1. An isolated and purified IL-9 mutein numbered in accordance with wild type IL-9 wherein said mutein is an IL-9 antagonist and comprises at least one amino-acid substitution, wherein said amino acid substitution is in the D helix at a position selected from the group consisting of 119-138.
2. The isolated and purified IL-9 mutein of claim 1 wherein said substitution is at a position selected from the group of consisting of 126, 133, 134 and 136.
3. The isolated and purified IL-9 mutein of claim 2 wherein said substitution at position 133 is Gln to Lys (SEQ ID NO. 21).
4. The isolated and purified IL-9 mutein of claim 2 wherein said substitution at position 134 is Lys to Glu (SEQ ID NO. 19).
5. The isolated and purified IL-9 mutein of claim 2 wherein said substitution at position 136 is Lys to Asp (SEQ ID NO. 17).
6. The IL-9 mutein of claim 1 , wherein said mutein inhibits the proliferative response of Mo7E cells to IL-9 with an IC50 value of about 0.1 nM to about 10 μM.
7. The IL-9 mutein of claim 1 , wherein said mutein inhibits the proliferative response of Mo7E cells to IL-9 with an IC50 value of about 0.5 nM to about 1 μM.
8. The IL-9 mutein of claim 1 , wherein said mutein inhibits the proliferative response of Mo7E cells to IL-9 with an IC50 value of about 1.0 nM to about 100 nM.
9. The IL-9 mutein of claim 1 , wherein said mutein inhibits the anti-apoptotic response of eosinophils to IL-9 with an IC50 value of about 0.1 nM to about 10 μM.
10. The IL-9 mutein of claim 1 , wherein said mutein inhibits the anti-apoptotic response of eosinophils to IL-9 with an IC50 value of about 0.5 nM to about 1 μM.
11. The IL-9 mutein of claim 1 , wherein said mutein inhibits the anti-apoptotic response of eosinophils to IL-9 with an IC50 value of about 1.0 nM to about 100 nM.
12. An isolated and purified IL-9 mutein numbered in accordance with wild type IL-9 wherein said mutein is an IL-9 antagonist and comprises at least one amino-acid substitution, wherein said substitution at position 126 is Lys to Glu and at 133 is Gln to Lys (SEQ ID NO. 24).