Soluble TNF Receptors and Their Use in Treatment of Disease

Abstract

The present invention relates to tumor necrosis factor (TNF) antagonists and corresponding nucleic acids derived from tumor necrosis factor receptors (TNFRs) and their use in the treatment of inflammatory diseases. These proteins are soluble secreted decoy receptors that bind to TNF and prevent TNF from signaling to cells. In particular, the proteins are mammalian TNFRs that lack exon 7 and which can bind TNF and can act as a TNF antagonist.

1 . An isolated protein capable of binding tumor necrosis factor (TNF), said protein having a sequence comprising the amino acids encoded by a cDNA derived from a mammalian tumor necrosis factor receptor (TNFR) gene, wherein the cDNA comprises in 5′ to 3′ contiguous order,
 the codon encoding the first amino acid after the cleavage point of the signal sequence of said gene through exon 6 of said gene and exon 8 or said gene through exon 10 of said gene; or 
 the codon encoding the first amino acid of the open reading frame of said gene through exon 6 of said gene and exon 8 of said gene through exon 10 of said gene. 
 
     
     
         2 . The protein of  claim 1 , wherein said TNF is TNF-α. 
     
     
         3 . The protein of  claim 1 , wherein said protein contains at least one processing, chemical, or post-translational modification, and wherein said modification is selected from the group consisting of acetylation, acylation, amidation, ADP-ribosylation, glycosylation, methylation, pegylation, prenylation, phosphorylation, or cholesterol conjugation. 
     
     
         4 . The protein of  claim 1 , wherein said receptor is TNFR1. 
     
     
         5 . The protein of  claim 4 , wherein said receptor is human TNFR1. 
     
     
         6 . The protein of  claim 1 , wherein said receptor is TNFR2. 
     
     
         7 . The protein of  claim 6 , wherein said receptor is human TNFR2. 
     
     
         8 . The protein of  claim 1 , wherein the sequence of said protein comprises a sequence selected from the group consisting of SEQ ID No:6, amino acids 30-417 of SEQ ID No:6, SEQ ID No:8, amino acids 30-416 of SEQ ID No:8, SEQ ID No:10, amino acids 23-435 of SEQ ID No:10, SEQ ID No:12, and amino acids 23-448 of SEQ ID No:12. 
     
     
         9 . A pharmaceutical composition comprising the protein of any one of  claim 1  or  7 , in admixture with a pharmaceutically acceptable carrier. 
     
     
         10 . A composition comprising the purified protein of  claim 1 . 
     
     
         11 . A method of treating an inflammatory disease or condition which comprises administering the pharmaceutical composition of  claim 9  to a subject for a time and in an amount effective to reduce the activity of TNF. 
     
     
         12 . The method of  claim 11 , wherein said disease or condition is selected from the group consisting of rheumatoid arthritis, juvenile rheumatoid arthritis, psoriasis, psoriatic arthritis, ankylosing spondylitis, inflammatory bowel disease (Crohn's disease or ulcerative colitis), hepatitis associated with hepatitis A virus, hepatitis associated with B virus, hepatitis associated with hepatitis C virus, hepatitis associated with ischemia/reperfusion, sepsis, alcoholic liver disease, and non-alcoholic steatosis. 
     
     
         13 . An isolated nucleic acid derived from a mammalian tumor necrosis factor receptor (TNFR) gene and encoding a protein capable of binding tumor necrosis factor (TNF), wherein the cDNA of said protein comprises in 5′ to 3′ contiguous order,
 the codon encoding the first amino acid after the cleavage point of the signal sequence of said gene through exon 6 of said gene and exon 8 of said gene through exon 10 of said gene; or 
 the codon encoding the first amino acid of the open reading frame of said gene through exon 6 of said gene and exon 8 of said gene through exon 10 of said gene. 
 
     
     
         14 . The nucleic acid of  claim 13 , wherein said receptor is TNFR1. 
     
     
         15 . The nucleic acid of  claim 14 , wherein said receptor is human TNFR1. 
     
     
         16 . The nucleic acid of  claim 13 , wherein said receptor is TNFR2. 
     
     
         17 . The nucleic acid of  claim 16 , wherein said receptor is human TNFR2. 
     
     
         18 . The nucleic acid of  claim 13 , wherein the sequence of said protein comprises a sequence selected from the group consisting of SEQ ID No:6, amino acids 30-417 of SEQ ID No:6, SEQ ID No:8, amino acids 30-416 of SEQ ID No:8, SEQ ID No:10, amino acids 23-435 of SEQ ID No:10, SEQ ID No:12, and amino acids 23-448 of SEQ ID No:12. 
     
     
         19 . The nucleic acid of  claim 16 , wherein the sequence of said nucleic acid comprises a sequence selected from the group consisting of nucleotides 1-1251 of SEQ ID No:5, nucleotides 88-1251 of SEQ ID No:5, nucleotides 1-1248 of SEQ ID No:7, nucleotides 88-1248 of SEQ ID No:7, nucleotides 1-1305 of SEQ ID No:9, nucleotides 67-1305 of SEQ ID No:9, nucleotides 1-1344 of SEQ ID No:11, and nucleotides 67-1344 of SEQ ID No:11. 
     
     
         20 . An expression vector comprising the nucleic acid of any one of  claim 13  or  17  operable linked to a regulatory sequence. 
     
     
         21 .- 56 .