Enhanced affinity hyaluronan binding peptides

Abstract

Novel hyaluronan-binding peptides are provided. The peptides are useful in preventing and treating disorders associated with altered tissue levels of hyaluronan or RHAMM, including cancer, inflammatory and autoimmune disorders and fibrotic disorders associated with tissue trauma.

I claim:  
     
         1 . A HA-binding peptide comprising: 
 (a) a sequence of the formula I:   X 1 -X 2 -X 1 -X 3 -X 4 -X 3 -X 4 -X 3 -X 3 -X 3 -X 5 -X 6 -X 6 -X 6 -X 1      wherein 
 each X 1  is independently selected from a hydroxy amino acid residue;  
 each X 2  is independently selected from a sulfur containing amino acid residue;  
 each X 3  is independently selected from a basic amino acid residue;  
 each X 4  is independently selected from an imino or aromatic amino acid residue;  
 each X 5  is independently selected from a dicarboxylic acid amino acid residue; and  
 each X 6  is independently selected from an aliphatic amino acid residue, and fragments, analogs or derivatives of the peptide which can bind HA;  
   (b) a sequence of the formula II:   Y 1 -Y 1 -Y 2 -Y 2 -Y 1 -Y 3 -Y 1 -Y 3 -Y 3 -Y 1 -Y 3 -Y 1 -Y 2 -Y 3 -Y 3      wherein 
 each Y 1  is independently selected from a hydroxy amino acid residue;  
 each Y 2  is independently selected from a sulfur containing amino acid residue; and  
 each Y 3  is independently selected from a basic amino acid residue, and fragments, analogs or derivatives of the peptide which bind HA; or  
   (c) a sequence of the formula III:   Z 1 -Z 1 -Z 2 -Z 2 -Z 1 -Z 3 -Z 1 -Z 3 -Z 3 -Z 1 -Z 3 -Z 1 -Z 3 -Z 3      wherein 
 each Z 1  is independently selected from a hydroxy amino acid residue;  
 each Z 2  is independently selected from a sulfur containing amino acid residue; and  
 each Z 3  is independently selected from a basic amino acid residue, and fragments, analogs or derivatives of the peptide which bind HA.  
   
     
     
         2 . A HA-binding peptide comprising a sequence of the formula I as defined in  claim 1  wherein 
 each X 1  is independently selected from threonine or serine;  
 each X 2  is independently selected from methionine or cysteine;  
 each X 3  is independently selected from arginine, lysine or histidine;  
 each X 4  is independently selected from proline, phenylalanine or tryptophan;  
 each X 5  is independently selected from asparagine or glutamine; and  
 each X 6  is independently selected from leucine, isoleucine, valine or alanine, and fragments, analogs or derivatives of the peptide which can bind HA.  
 
     
     
         3 . A peptide according to  claim 2  comprising the amino acid sequence TMTRPHFHKRQLVLS.  
     
     
         4 . A peptide according to  claim 3  wherein the amino acids in the peptide are the levorotatory (L) form.  
     
     
         5 . A HA-binding peptide comprising a sequence of the Formula II as defined in  claim 1 , wherein 
 each Y 1  is independently selected from serine or threonine;    each Y 2  is independently selected from methionine or cysteine; and    each Y 3  is independently selected from arginine, lysine or histidine, and fragments, analogs or derivatives of the peptide which bind HA.    
     
     
         6 . A peptide according to  claim 5  comprising the amino acid sequence STMMSRSHKTRSCHH.  
     
     
         7 . A peptide according to  claim 6  wherein the amino acids in the peptide are the levorotatory (L) form.  
     
     
         8 . A HA-binding peptide comprising a sequence of the formula III as defined in  claim 1 , wherein 
 each Z 1  is independently selected from serine or threonine;    each Z 2  is independently selected from methionine or cysteine; and    each Z 3  is independently selected from arginine, lysine or histidine, and fragments, analogs or derivatives of the peptide which bind HA.    
     
     
         9 . A peptide according to  claim 8  comprising the amino acid sequence STMMSRSHKTRSHH.  
     
     
         10 . A peptide according to  claim 9  wherein the amino acids in the peptide are the levorotatory (L) form.  
     
     
         11 . A peptide according to  claim 8  comprising the amino acid sequence STMMSRSHKTRSHHV.  
     
     
         12 . A peptide according to  claim 11  wherein the amino acids in the peptide are the levorotatory (L) form.  
     
     
         13 . An isolated nucleic acid molecule encoding a HA binding peptide according to  claim 1 .  
     
     
         14 . An isolated nucleic acid molecule according to  claim 13  encoding a HA binding peptide and comprising a nucleotide sequence selected from the group consisting of: 
 (a) the nucleotide sequence shown in SEQ ID NO. 5;  
 (b) the nucleotide sequence shown in SEQ ID NO. 6;  
 (c) the nucleotide sequence shown in SEQ ID NO. 7;  
 (d) the nucleotide sequence shown in SEQ ID NO. 8;  
 (e) the nucleotide sequence shown in SEQ ID NO. 9;  
 (f) the nucleotide sequence shown in SEQ ID NO. 10;  
 (g) the nucleotide sequence shown in SEQ ID NO. 11; and  
 (h) the nucleotide sequence shown in SEQ ID NO. 12.  
 
     
     
         15 . An expression vector comprising an isolated nucleic acid molecule according to  claim 13  and regulatory sequences suitable for expression of the nucleic acid molecule.  
     
     
         16 . A method of modulating cell locomotion comprising administering an effective amount of one or more hyaluronan-binding peptides according to  claim 1  to a cell or animal in need thereof.  
     
     
         17 . A method of modulating cell locomotion comprising administering an effective amount of hyaluronan-binding peptide according to  claim 3  to a cell or animal in need thereof.  
     
     
         18 . A method of modulating cell locomotion comprising administering an effective amount of hyaluronan-binding peptide according to  claim 6  to a cell or animal in need thereof.  
     
     
         19 . A method of modulating cell locomotion comprising administering an effective amount of hyaluronan-binding peptide according to  claim 9  to a cell or animal in need thereof.  
     
     
         20 . A method of modulating cell locomotion comprising administering an effective amount of hyaluronan-binding peptide according to  claim 11  to a cell or animal in need thereof.  
     
     
         21 . A method of preventing or inhibiting tissue fibrosis comprising administering an effective amount of one or more hyaluronan-binding peptides according to  claim 1  to an animal in need thereof.  
     
     
         22 . A method of preventing or inhibiting tissue fibrosis comprising administering an effective amount of hyaluronan-binding peptide according to  claim 3  to an animal in need thereof.  
     
     
         23 . A method of preventing or inhibiting tissue fibrosis comprising administering an effective amount of hyaluronan-binding peptide according to  claim 6  to an animal in need thereof.  
     
     
         24 . A method of preventing or inhibiting tissue fibrosis comprising administering an effective amount of hyaluronan-binding peptide according to  claim 9  to an animal in need thereof.  
     
     
         25 . A method of preventing or inhibiting tissue fibrosis comprising administering an effective amount of hyaluronan-binding peptide according to  claim 11  to an animal in need thereof.  
     
     
         26 . A method according to  claim 21  wherein the tissue fibrosis is caused by a disorder selected from the group consisting of tissue malfunction due to keloids, hypertrophic scars, anatomonic strictures, intra-abdominal adhesions, cirrhosis of the liver, neurological deficits following spinal cord injury, valvular heart diseases, burn-injured joints, failure of anastomosis and adhesions following surgery.  
     
     
         27 . A method of treating or preventing cancer comprising administering an effective amount of one or more hyaluronan-binding peptides according to  claim 1  to an animal in need thereof.  
     
     
         28 . A method of preventing or reducing the metastasis of cancer cells comprising administering an effective amount of one or more hyaluronan-binding peptides according to  claim 1  to an animal in need thereof.