Protein based TNF-α variants for the treatment of TNF-α related disorders
The invention relates to novel proteins with TNF-α antagonist activity and nucleic acids encoding these proteins. The invention further relates to the use of the novel proteins in the treatment of TNF-α related disorders.
1. A method of selectively inhibiting the activity of wild-type soluble Tumor necrosis factor-α (TNF-α) in a human comprising administering to said human a molecule that inhibits the activity of soluble TNF-α while substantially maintaining the activity of transmembrane TNF-α, wherein said molecule is a variant TNF-α as compared to human wild-type TNF-α (SEQ ID NO: 12), wherein said TNF-α variant comprises the amino acid modifications Y87H and A145R or I97T and A145R.
2. A method of claim 1 , wherein said TNF-α variant is substantially free of agonistic activity.
3. A method of selectively inhibiting the activity of wild-type soluble TNF-α as compared to the activity of transmembrane wild-type TNF-α in a mammal comprising administering to said mammal a variant TNF-α molecule as compared to the wild-type human TNF-α of SEQ ID NO:12, wherein said TNF-α variant is substantially free of agonistic activity, wherein said TNF-α variant comprises the amino acid modifications Y87H and A145R or I97T and A145R.
4. A method of forming a TNF-α heterotrimer comprising contacting a variant TNF-α molecule as compared to the wild-type human TNF-α of SEQ ID NO:12 with the wild type TNF-α, under conditions whereby a heterotrimer is formed, wherein said TNF-α variant is substantially free of agonistic activity, wherein said TNF-α variant comprises the amino acid modifications Y87H and A145R or I97T and A145R.
5. A method according to claim 2 , 3 or 4 , wherein the agonistic activity of said TNF-α variant is reduced by at least 50%.
6. A method of claim 1 , wherein said TNF-α variant comprises the amino acid modifications Y87H and A145R.
7. A method of claim 1 , wherein said TNF-α variant comprises the amino acid modifications I97T and A145R.
8. A method of claim 6 or 7 , wherein said variant further comprises the amino acid modification R31C.
9. A method according to claim 8 , wherein said variant TNF-α comprises polyethylene glycol.
10. A method according to claim 8 , wherein said variant further comprises an amino acid modification at position 101.
11. A method of claim 10 , wherein said modification is C101A.
12. A method of claim 6 or 7 , wherein said variant further comprises the amino acid modification V1M.
13. A method according to claim 12 , wherein said variant further comprises an amino acid modification at position 101.
14. A method of claim 13 , wherein said modification is C101A.
15. A method of claim 6 or 7 , wherein said variant further comprises an amino acid modification at position 69.
16. A method of claim 15 , wherein said modification is C69V.
17. A method of claim 6 or 7 , wherein said variant further comprises an amino acid modification at position 101.
18. A method of claim 17 , wherein said modification is C101A.
19. A method of claim 1 , wherein said variant TNF-α comprises the amino acid substitutions V1M, R31C, C69V, Y87H, C101, and A145R.
20. A method of claim 1 wherein said TNF-α variant is selected from the group consisting of XENP268, XENP344, XENP345, XENP346, XENP550, XENP551, XENP557, XENP1593, XENP1594, and XENP1595.