Single-Chain Trail-Receptor Agonist Proteins

Abstract

Provided herein are specific TRAIL receptor agonist proteins, nucleic acids encoding the same, and methods of treating a subject having a TRAIL-associated disease or disorder. The TRAIL receptor agonist proteins provided herein comprise three soluble TRAIL domains and an Fc fragment. The TRAIL receptor agonist proteins are substantially non-aggregating and suitable for therapeutic, diagnostic and/or research applications.

1 . A TRAIL receptor agonist protein comprising a polypeptide having the amino acid sequence set forth in SEQ ID NO: 19. 
     
     
         2 . A TRAIL receptor agonist protein comprising two polypeptides having the amino acid sequence set forth in SEQ ID NO: 19. 
     
     
         3 . The TRAIL receptor agonist protein of  claim 2 , wherein the two polypeptides are covalently linked through three interchain disulfide bonds formed between cysteine residues 513, 519, and 522 of each polypeptide. 
     
     
         4 . The TRAIL receptor agonist protein of  claim 2 , wherein one or more of the asparagine residues at positions 168 and 337 of the polypeptide(s) are N-glycosylated. 
     
     
         5 . The TRAIL receptor agonist protein of  claim 2 , wherein the asparagine residues at positions 168 and 337 of the polypeptide(s) are both N-glycosylated. 
     
     
         6 . The TRAIL receptor agonist protein of  claim 5 , wherein the polypeptide(s) are further post-translationally modified. 
     
     
         7 . The TRAIL receptor agonist protein of  claim 6 , wherein the post-translational modification comprises modification of the N-terminal glutamine to pyroglutamate. 
     
     
         8 . A pharmaceutical composition comprising the TRAIL receptor agonist protein of any one of  claim 6  and one or more pharmaceutically acceptable carriers, diluents, excipients, and/or adjuvants. 
     
     
         9 . A nucleic acid molecule encoding the TRAIL receptor agonist protein of  claim 1 . 
     
     
         10 . An expression vector comprising the nucleic acid molecule of  claim 9 . 
     
     
         11 . A cell comprising the nucleic acid molecule of  claim 9 . 
     
     
         12 . The cell of  claim 11 , which is a eukaryotic cell. 
     
     
         13 . The cell of  claim 11 , wherein the cell is a mammalian cell. 
     
     
         14 . The cell of  claim 11 , wherein the cell is a Chinese Hamster Ovary (CHO) cell. 
     
     
         15 . A method of treating a subject having a TRAIL-associated disease or disorder, the method comprising administering to the subject an effective amount of the TRAIL receptor agonist protein of any one of  claim 6 . 
     
     
         16 . The method of  claim 15 , wherein the disease or disorder is selected from the group consisting of: tumors, infectious diseases, inflammatory diseases, metabolic diseases, autoimmune disorders, degenerative diseases, apoptosis-associated diseases, and transplant rejections. 
     
     
         17 . The method of  claim 16 , wherein the tumors are solid tumors. 
     
     
         18 . The method of  claim 16 , wherein the tumors are lymphatic tumors. 
     
     
         19 . The method of  claim 16 , wherein the autoimmune disorders are rheumatoid diseases, arthritic diseases, or rheumatoid and arthritic diseases. 
     
     
         20 . The method of  claim 16 , wherein the disease or disorder is rheumatoid arthritis. 
     
     
         21 . The method of  claim 16 , wherein the degenerative disease is a neurodegenerative disease. 
     
     
         22 . The method of  claim 16 , wherein the neurodegenerative disease is multiple sclerosis.