Protein based TNF-alpha variants for the treatment of TNF-alpha related disorders

Abstract

The invention relates to novel proteins with TNF-alpha antagonist activity and nucleic acids encoding these proteins. The invention further relates to the use of the novel proteins in the treatment of TNF-alpha related disorders.

1 .- 28 . (canceled)  
     
     
         29 . A method of antagonizing soluble TNF-α activity without significantly altering the activity of transmembrane TNF-α comprising administering a PEGylated TNF-α to a patient.  
     
     
         30 . The method of  claim 29 , wherein said TNF-α comprises a cysteine amino acid at an amino acid position selected from the group consisting of 10, 21, 23, 24, 25, 27, 31, 42, 44, 45, 46, 86, 87, 88, 90, 107, 108, 128, 110, 140 and 145, and said PEG moiety is attached at said cysteine amino acid.  
     
     
         31 . The method of  claim 30  wherein said position is 31.  
     
     
         32 . The method of  claim 30  wherein said TNF-α further comprises at least one amino acid substitution at a position selected from the group consisting of 57, 75, 86, 87, 97, 115, 143, 145, and 146.  
     
     
         33 . The method of  claim 29  wherein said PEG is PEG-10.  
     
     
         34 . The method of  claim 29  wherein said PEG is PEG-5.  
     
     
         35 . The method of  claim 30  wherein said substitutions are selected from the group consisting of Q21C, Q21R, E23C, N34E, V91E, Q21R, N30D, R31C, R31I, R31D, R31E, R32D, R32E, R32S, A33E, N34E, N34V, A35S, D45C, L57F, L57W, L57Y, K65D, K65E, K651, K65M, K65N, K65Q, K65T, K65S, K65V, K65W, G66K, G66Q, Q67D, Q67K, Q67R, Q67S, Q67W, Q67Y, L75E, L75K, L75Q, A84V, S86Q, S86R, Y87H, Y87R, V91E, I97R, I97T, A111R, A111E, K112D, K112E, Y115D, Y115E, Y115F, Y115H, Y115I, Y115K, Y115L, Y115M, Y115N, Y115Q, Y115R, Y115S, Y115T, Y115W, D140K, D140R, D143E, D143K, D143L, D143R, D143N, D143Q, D143R, D143S, F144N, A145D, A145E, A145F, A145H, A145K, A145M, A145N, A145Q, A145R, A145S, A145T, A145Y, E146K, E146L, E146M, E146N, E146R, E146S and S147R.  
     
     
         36 . A method according to  claim 29  wherein said method results in a reduced infection rate in said patient as compared to a patient administered a TNF-alpha without a PEG moiety.  
     
     
         37 . A method of antagonizing soluble TNF-α activity without significantly altering the activity of transmembrane TNF-α comprising administering a TNF-α variant to a patient, said variant comprising an amino acid substitution as compared to amino acid nos 8-164 of SEQ ID NO:X at a position selected from positions 21, 23, 30, 31, 32, 33, 34, 35, 57, 65, 66, 67, 75, 84, 86, 87, 91, 97, 111, 112, 115, 140, 143, 144, 145, 146 and 147.