Single-chain light receptor agonist proteins

Abstract

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

1 . A LIGHT receptor agonist protein comprising a single-chain fusion polypeptide comprising:
 (i) a first soluble LIGHT cytokine domain,   (ii) a first peptide linker,   (iii) a second soluble LIGHT cytokine domain,   (iv) a second peptide linker, and   (v) a third soluble LIGHT cytokine domain,   (vi) a hinge-linker, and   (vii) an antibody Fc fragment, wherein the antibody Fc fragment (vii) consists of the amino acid sequence as shown in SEQ ID NO: 13 or amino acids 1-217 of SEQ ID NO: 13;
 wherein the soluble LIGHT cytokine domains (i), (iii) and (v) each consists of amino acids E91-V240, or N93-V240, or P94-V240 of human LIGHT cytokine according to SEQ ID NO: 1. 
   
     
     
         2 . The LIGHT receptor agonist protein of  claim 1 , wherein the antibody Fc fragment (vii) is fused to the C-terminal end of the third LIGHT cytokine domain (v) via the hinge-linker (vi). 
     
     
         3 . The LIGHT receptor agonist protein of  claim 1 , wherein the soluble LIGHT cytokine domains (i), (iii) and (v) each consists of amino acids P94-V240 of human LIGHT cytokine according to SEQ ID NO: 1. 
     
     
         4 . The LIGHT receptor agonist protein of  claim 1 , wherein the soluble LIGHT cytokine domain (i) consists of amino acids E91-V240 of SEQ ID:1, and wherein the soluble LIGHT cytokine domains (iii) and (v) consist of either amino acid N93-V240 or P94-V240. 
     
     
         5 . The LIGHT receptor agonist protein of  claim 1 , which is substantially non-aggregating. 
     
     
         6 . The LIGHT receptor agonist protein of  claim 1 , wherein the first and second peptide linkers (ii) and (iv) independently consist of glycine and serine, or consist of glycine, serine and asparagine, wherein the asparagine is optionally glycosylated. 
     
     
         7 . The LIGHT receptor agonist protein of  claim 6 , wherein the first and the second peptide linkers (ii) and (iv) comprise the amino acid sequence according to SEQ ID NO: 2. 
     
     
         8 . The LIGHT receptor agonist protein of  claim 1 , wherein the hinge linker has one of the sequences of SEQ ID NOs: 16 and 19-24. 
     
     
         9 . The LIGHT receptor agonist protein of  claim 3 , wherein the hinge linker has one of the sequences of SEQ ID NOs: 16 and 19-24. 
     
     
         10 . The LIGHT receptor agonist protein of  claim 4 , wherein the hinge linker has one of the sequences of SEQ ID NOs: 16 and 19-24. 
     
     
         11 . The LIGHT receptor agonist of  claim 1 , comprising two of the single-chain fusion polypeptides. 
     
     
         12 . The LIGHT receptor agonist protein of  claim 11 , wherein the polypeptide(s) are further post-translationally modified. 
     
     
         13 . The LIGHT receptor agonist protein of  claim 12 , wherein the post-translational modification comprises modification of the N-terminal glutamine to pyroglutamate. 
     
     
         14 . A pharmaceutical or diagnostic composition comprising the LIGHT receptor agonist protein of  claim 1  and a pharmaceutically acceptable carriers, diluents, excipients, and/or adjuvants. 
     
     
         15 . A LIGHT receptor agonist protein comprising a single-chain fusion polypeptide comprising:
 (i) a first soluble LIGHT cytokine domain,   (ii) a first peptide linker,   (iii) a second soluble LIGHT cytokine domain,   (iv) a second peptide linker, and   (v) a third soluble LIGHT cytokine domain,   (vi) a hinge-linker, and   (vii) an antibody Fc fragment, wherein the antibody Fc fragment (vii) consists of the amino acid sequence as shown in SEQ ID NO: 13 or amino acids 1-217 of SEQ ID NO: 13;   
       wherein the soluble LIGHT cytokine domains (i), (iii) and (v) each independently consists of amino acids E91-V240, or N93-V240, or P94-V240 of human LIGHT cytokine according to SEQ ID NO: 1, having no mutation or having one or two mutations at position selected from the group consisting of: N93, N102, E115, T116, Q117, L118, G119, L120, C154, R172, Y173, E175, E176, E178, C187, R228, D229. 
     
     
         16 . The LIGHT receptor agonist protein of  claim 15 , wherein N93 is replaced by glycine or serine in at least one of the soluble LIGHT cytokine domains. 
     
     
         17 . The LIGHT receptor agonist protein of  claim 15 , wherein P94 is replaced by glycine or serine in at least one of the soluble LIGHT cytokine domains. 
     
     
         18 . The LIGHT receptor agonist protein of  claim 15 , wherein E91 is replaced by glutamine, glycine, or serine. 
     
     
         19 . The LIGHT receptor agonist protein of  claim 15 , wherein the first and second peptide linkers (ii) and (iv) independently consist of glycine and serine, or consist of glycine, serine and asparagine, wherein the asparagine is optionally glycosylated. 
     
     
         20 . The LIGHT receptor agonist protein of  claim 15 , wherein the first and the second peptide linkers (ii) and (iv) consist of the amino acid sequence according to SEQ ID NO: 2. 
     
     
         21 . The LIGHT receptor agonist protein of  claim 15 , wherein the hinge linker has one of the sequences of SEQ ID NOs: 16 and 19-24.