Modified IL-4 mutein receptor antagonists
This invention relates to modified IL-4 mutein receptor antagonists comprising an IL-4 mutein receptor antagonist coupled to polyethylene glycol. Related formulations and dosages and methods of administration thereof for therapeutic purposes are also provided. These modified IL-4 mutein receptor antagonists, compositions and methods provide a treatment option for those individuals afflicted with a respiratory disorder such as asthma by inhibiting IL-4 and IL-13-mediated airway hyperresponsiveness and eosinophilia. More particularly, these antagonists have an increased duration of effect versus unmodified IL-4RA by virtue of a greater plasma half-life.
1. A modified IL-4 mutein receptor antagonist wherein the amino acid residue at position 37 , 38 , or 104 is cysteine and produced by
a) culturing a host cell comprising an expression vector comprising a polynucleotide sequence as set forth in SEQ ID NO: 4, SEQ ID NO: 5, or SEQ ID NO: 6; and
b) purifying the antagonist from the host cell culture, wherein the antagonist inhibits IL-4 and IL-13-mediated activity.
2. The modified IL-4 mutein receptor antagonist of claim 1 coupled to a non-protein polymer selected from the group consisting of polyethylene glycol, polypropylene glycol and polyoxyalkylenes.
3. The modified IL-4 mutein receptor antagonist of claim 2 wherein the modified mutein receptor antagonist binds to the IL-4 receptor alpha chain with a K d of about 0.1 nM to about 10 μM, about 0.5 nM to about 1 μM, or about 1.0 nM to about 100 nM.
4. The modified IL-4 mutein receptor antagonist of claim 2 wherein the modified IL-4 mutein receptor antagonist inhibits the proliferative response of TF-1 cells to IL-4 with an IC 50 of about 0.1 nM to about 10 μM, about 0.5 nM to about 1 μM, or about 1.0 nM to about 100 nM.
5. The modified IL-4 mutein receptor antagonist of claim 2 wherein the modified IL-4 mutein receptor antagonist inhibits the proliferative response of TF-1 cells to IL-13 with an IC 50 of about 0.1 nM to about 10 μM, about 0.5 nM to about 1 μM, or about 1.0 nM to about 100 nM.
6. The modified IL-4 mutein receptor antagonist of claim 2 wherein the modified IL-4 mutein receptor antagonist inhibits the proliferative response of human B cells to IL-4 with an IC 50 of about 0.1 nM to about 10 μM, about 0.5 nM to about 1 μM, or about 1.0 nM to about 100 nM.
7. The modified IL-4 mutein receptor antagonist of claim 2 wherein the modified IL-4 mutein receptor antagonist inhibits the proliferative response of human T cells to IL-4 with an IC 50 of about 0.1 nM to about 10 μM, about 0.5 nM to about 1 μM, or about 1.0 nM to about 100 nM.
8. The modified IL-4 mutein receptor antagonist of claim 2 wherein the modified IL-4 mutein receptor antagonist has a plasma half-life which is at least about 2-10 fold greater than that of an unmodified IL-4 receptor antagonist.
9. A pharmaceutical composition comprising:
a) the modified IL-4 mutein receptor antagonist of claim 2 ; and
b) a pharmaceutically acceptable carrier.
10. A modified IL-4 mutein receptor antagonist coupled to a non-protein polymer at an amino acid residue at position 37 , 38 , or 104 of IL-4, wherein the amino acid at 37 , 38 or 104 is cysteine, and wherein the non-protein polymer is polyethylene glycol, polypropylene glycol or a polyoxyalkylene.
11. The modified IL-4 mutein receptor antagonist of claim 2 or 10 comprising the amino acid sequence as set forth in SEQ ID NO: 12.
12. The modified IL-4 mutein receptor antagonist of claim 2 or 10 comprising the amino acid sequence as set forth in SEQ ID NO: 13.
13. The modified IL-4 mutein receptor antagonist of claim 2 or 10 comprising an the amino acid sequence as set forth in SEQ ID NO: 14.
14. The modified IL-4 mutein receptor antagonist of claim 10 wherein the modified mutein receptor antagonist binds to the IL-4 receptor alpha chain with a K d of about 0.1 nM to about 10 μM, about 0.5 nM to about 1 μM, or about 1.0 nM to about 100 nM.
15. The modified IL-4 mutein receptor antagonist of claim 10 wherein the modified IL-4 mutein receptor antagonist inhibits the proliferative response of TF-1 cells to IL-4 with an IC 50 of about 0.1 nM to about 10 μM, about 0.5 nM to about 1 μM, or about 1.0 nM to about 100 nM.
16. The modified IL-4 mutein receptor antagonist of claim 10 wherein the modified IL-4 mutein receptor antagonist inhibits the proliferative response of TF-1 cells to IL-13 with an IC 50 of about 0.1 nM to about 10 μM, about 0.5 nM to about 1 μM, or about 1.0 nM to about 100 nM.
17. The modified IL-4 mutein receptor antagonist of claim 10 wherein the modified IL-4 mutein receptor antagonist inhibits the proliferative response of human B cells to IL-4 with an IC 50 of about 0.1 nM to about 10 μM, about 0.5 nM to about 1 μM, or about 1.0 nM to about 100 nM.
18. The modified IL-4 mutein receptor antagonist of claim 10 wherein the modified IL-4 mutein receptor antagonist inhibits the proliferative response of human T cells to IL-4 with an IC 50 of about 0.1 nM to about 10 μM, about 0.5 nM to about 1 μM, or about 1.0 nM to about 100 nM.
19. The modified IL-4 mutein receptor antagonist of claim 10 wherein the modified IL-4 mutein receptor antagonist has a plasma half-life which is at least about 2-10 fold greater than that of an unmodified IL-4 receptor antagonist.
20. A pharmaceutical composition comprising:
a) the modified IL-4 mutein receptor antagonist of claim 10 ; and
b) a pharmaceutically acceptable carrier.
21. A modified IL-4 mutein receptor antagonist wherein the amino acid at 37 , 38 and 104 is cysteine, and produced by
a) culturing a host cell comprising an expression vector comprising a polynucleotide sequence as set forth in SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, wherein the antagonist is expressed;
b) allowing the antagonist to refold in the presence of dithiothreitol; and
c) purifying the antagonist from the host cell culture, wherein the antagonist inhibits IL-4 and IL-13-mediated activity.
22. The modified IL-4 mutein receptor antagonist of claim 21 coupled to a non-protein polymer selected from the group consisting of:
polyethylene glycol, polypropylene glycol or a polyoxyalkylene.
23. The modified IL-4 mutein receptor antagonist of claim 22 wherein the modified mutein receptor antagonist binds to the IL-4 receptor alpha chain with a K d of about 0.1 nM to about 10 μM, about 0.5 nM to about 1 μM, or about 1.0 nM to about 100 nM.
24. The modified IL-4 mutein receptor antagonist of claim 22 wherein the modified IL-4 mutein receptor antagonist inhibits the proliferative response of TF-1 cells to IL-4 with an IC 50 of about 0.1 nM to about 10 μM, about 0.5 nM to about 1 μM, or about 1.0 nM to about 100 nM.
25. The modified IL-4 mutein receptor antagonist of claim 22 wherein the modified IL-4 mutein receptor antagonist inhibits the proliferative response of TF-1 cells to IL-13 with an IC 50 of about 0.1 nM to about 10 μM, about 0.5 nM to about 1 μM, or about 1.0 nM to about 100 nM.
26. The modified IL-4 mutein receptor antagonist of claim 22 wherein the modified IL-4 mutein receptor antagonist inhibits the proliferative response of human B cells to IL-4 with an IC 50 of about 0.1 nM to about 10 μM, about 0.5 nM to about 1 μM, or about 1.0 nM to about 100 nM.
27. The modified IL-4 mutein receptor antagonist of claim 22 wherein the modified IL-4 mutein receptor antagonist inhibits the proliferative response of human T cells to IL-4 with an IC 50 of about 0.1 nM to about 10 μM, about 0.5 nM to about 1 μM, or about 1.0 nM to about 100 nM.
28. The modified IL-4 mutein receptor antagonist of claim 22 wherein the modified IL-4 mutein receptor antagonist has a plasma half-life which is at least about 2-10 fold greater than that of an unmodified IL-4 receptor antagonist.
29. A pharmaceutical composition comprising:
a) the modified IL-4 mutein receptor antagonist of claim 21 ; and
b) a pharmaceutically acceptable carrier.
30. The modified IL-4 mutein receptor antagonist of claim 22 , wherein the non-protein polymer is polyethylene glycol (PEG).