OX40R Binding Agents

Abstract

The present invention discloses peptides isolated from the extracellular domain of OX40 Ligand (OX40L) capable of binding OX40 Receptor (OX40R) and inhibiting OX40R-OX40L interaction. Such peptides, fusion proteins comprising them, as well as peptides and other molecules designed on their sequences, can be used as OX40R binding agents competing with natural OX40L for blocking OX40R-mediated cell signaling in the prophylaxis and/or treatment of diseases related to activated T cells.

1 . A screening assay for compounds modulating OX40R-OX40L interactions comprising:
 a) forming a sample comprising the following elements:
 i) an element constituting an OX40R binding agent; 
 ii) an element constituting the OX40R moiety, chosen from a protein comprising the extracellular domain of OX40R, a cell line expressing OX40R extracellular domain on its surface, or a cell line secreting extracellular domain of OX40R; and 
 iii) compound(s) to be tested as modulator(s) OX40R-OX40L interactions; 
   b) detecting, directly or indirectly, the effect of the compounds (iii) on the interactions between the elements (i) and (ii); and   c) comparing the effect detected in (b) amongst samples different in terms of quality, quantity or both quality and quantity of the elements of (a), wherein said OX40R binding agent is selected from the group consisting of:
 a) SEQ ID NO: 6; 
 b) SEQ ID NO: 6, wherein one or more amino acids have been deleted, said polypeptide contains SEQ ID NO: 13 and said polypeptide binds to the OX40 receptor (OX40R); 
 c) between 5 and 10 contiguous amino acids of SEQ ID NO: 1, wherein said polypeptide contains SEQ ID NO: 13 and binds to OX40R; 
 d) SEQ ID NO: 8 or SEQ ID NO: 13; 
 e) an active mutant of a), b), c) or d), wherein one or more of the amino acids has been conservatively substituted and said active mutant binds to OX40R; 
 f) a fusion polypeptide or peptide comprising a first amino acid sequence and a second amino acid sequence, wherein said first amino acid sequence is a protein sequence other than human OX40L fused to a second amino acid sequence selected from:
 i) SEQ ID NO: 6; 
 ii) SEQ ID NO: 6, wherein one or more amino acids have been deleted, said second amino acid sequence contains SEQ ID NO: 13 and said fusion polypeptide hinds to the OX40 receptor (OX40R); 
 iii) between 5 and 10 contiguous amino acids of SEQ ID NO: 1, wherein said contiguous amino acids of SEQ ID NO: 1 contain SEQ ID NO: 13 and said fusion polypeptide binds to OX40R; or 
 iv) SEQ ID NO: 8 or SEQ ID NO: 13; and 
 
 g) a conjugate or derivative of a), b), c), d), e) or f). 
   
     
     
         2 . The screening assay according to  claim 1 , wherein said polypeptide is attached to a solid support. 
     
     
         3 . A method of treating a disease or condition comprising the administration of to an individual in need of treatment a composition comprising a polypeptide consisting of:
 a) SEQ ID NO: 6;   b) SEQ ID NO: 6, wherein one or more amino acids have been deleted, said polypeptide contains SEQ ID NO: 13 and said polypeptide binds to the OX40 receptor (OX40R);   c) between 5 and 10 contiguous amino acids of SEQ ID NO: 1, wherein said polypeptide contains SEQ ID NO: 13 and binds to OX40R;   d) SEQ ID NO: S or SEQ ID NO: 13;   e) an active mutant of a), b), c) or d), wherein one or more of the amino acids has been conservatively substituted and said active mutant binds to OX40R;   f) a fusion polypeptide or peptide comprising a first amino acid sequence and a second amino acid sequence, wherein said first amino acid sequence is a protein sequence other than human OX40L fused to a second amino acid sequence selected from:
 i) SEQ ID NO: 6; 
 ii) SEQ ID NO: 6, wherein one or more amino acids have been deleted, said second amino acid sequence contains SEQ ID NO: 13 and said fusion polypeptide binds to the OX40 receptor (OX40R); 
 iii) between 5 and 10 contiguous amino acids of SEQ ID NO: 1, wherein said contiguous amino acids of SEQ ID NO: 1 contain SEQ ID NO: 13 and said fusion polypeptide binds to OX40R; or 
 iv) SEQ ID NO: 8 or SEQ ID NO: 13; or 
   g) a conjugate or derivative of a), b), c), d), e) or f).   
     
     
         4 . The method according to  claim 3 , wherein said disease or condition is autoimmune disease inflammations or infection where an inhibition of CD4 +  T cells activation is beneficial. 
     
     
         5 . The method according to  claim 4 , wherein said disease or condition is a tumor, inflammatory bowel disease, rheumatoid arthritis or multiple sclerosis. 
     
     
         6 . A method of detecting OX40R comprising contacting a sample with an OX40R binding agent selected from the group consisting of:
 a) SEQ ID NO: 6;   b) SEQ ID NO: 6, wherein one or more amino acids have been deleted, said polypeptide contains SEQ ID NO: 13 and said polypeptide binds to the OX40 receptor (OX40R);   c) between 5 and 10 contiguous amino acids of SEQ ID NO: 1, wherein said polypeptide contains SEQ ID NO: 13 and binds to OX40R;   d) SEQ ID NO: 8 or SEQ ID NO: 13;   e) an active mutant of a), b), c) or d), wherein one or more of the amino acids has been conservatively substituted and said active mutant binds to OX40R;   f) a fusion polypeptide or peptide comprising a first amino acid sequence and a second amino acid sequence, wherein said first amino acid sequence is a protein sequence other than human OX40L fused to a second amino acid sequence selected from:
 i) SEQ ID NO: 6; 
 ii) SEQ ID NO: 6, wherein one or more amino acids have been deleted, said second amino acid sequence contains SEQ ID NO: 13 and said fusion polypeptide binds to the OX40 receptor (OX40R); 
 iii) between 5 and 10 contiguous amino acids of SEQ ID NO: 1, wherein said contiguous amino acids of SEQ ID NO: 1 contain SEQ ID NO: 13 and said fusion polypeptide binds to OX40R; or 
 iv) SEQ ID NO: 8 or SEQ ID NO: 13; and 
   g) a conjugate or derivative of a), b), c), d), e) or f).   
     
     
         7 . The method according to  claim 6 , wherein said binding agent is attached to a solid support. 
     
     
         8 . The method according to  claim 6 , wherein said sample contains activated CD4′ T cells. 
     
     
         9 . The method according to  claim 6 , wherein said sample contains the extracellular domain of OX40R protein as membrane-bound or a soluble protein. 
     
     
         10 . A method of antagonizing the activity of OX40L comprising contacting a composition containing OX40R with a composition comprising a polypeptide consisting of:
 a) SEQ ID NO: 6;   b) SEQ ID NO: 6, wherein one or more amino acids have been deleted, said polypeptide contains SEQ ID NO: 13 and said polypeptide binds to the OX40 receptor (OX40R);   c) between 5 and 10 contiguous amino acids of SEQ ID NO: 1, wherein said polypeptide contains SEQ ID NO: 13 and binds to OX40R;   d) SEQ ID NO: 8 or SEQ ID NO: 13;   e) an active mutant of a), b), c) or d), wherein one or more of the amino acids has been conservatively substituted and said active mutant hinds to OX40R;   f) a fusion polypeptide or peptide comprising a first amino acid sequence and a second amino acid sequence, wherein said first amino acid sequence is a protein sequence other than human OX40L fused to a second amino acid sequence selected from:
 i) SEQ ID NO: 6; 
 ii) SEQ ID NO: 6, wherein one or more amino acids have been deleted, said second amino acid sequence contains SEQ ID NO: 13 and said fusion polypeptide binds to the OX40 receptor (OX40R); 
 iii) between 5 and 10 contiguous amino acids of SEQ ID NO: 1, wherein said contiguous amino acids of SEQ ID NO: 1 contain SEQ ID NO: 13 and said fusion polypeptide binds to OX40R; or 
 iv) SEQ ID NO: 8 or SEQ ID NO: 13; or 
   g) a conjugate or derivative of a), b), c), d), e) or f).