Anti-invasive and anti-angiogenic compositions

Abstract

A peptide compound having the sequence Lys-Pro-Ser-Ser-Pro-Pro-Glu-Glu [SEQ ID NO:2] or a substitution variant, addition variant or other chemical derivative thereof inhibits cell invasion, endothelial tube formation or angiogenesis in vitro. A number of substitution variants and addition variants of this peptide, preferably capped at the N- and C-termini, as well as peptidomimetic derivatives, are useful for treating diseases and conditions mediated by undesired and uncontrolled cell invasion and/or angiogenesis. Pharmaceutical compositions comprising the above peptides and derivatives are administered to subjects in need of such treatment in a dosage sufficient to inhibit invasion and/or angiogenesis. The disclosed compositions and methods are particularly useful for suppressing the growth and metastasis of tumors.

What is claimed is:  
     
         1 . A peptide compound having the sequence  
       Lys-Pro-Ser-Ser-Pro-Pro-Glu-Glu  [SEQ ID NO:2] 
       or a substitution variant, addition variant or other chemical derivative thereof, which peptide, variant or derivative is capped or uncapped, 
 wherein said peptide, variant or derivative has one or more of the following activities: 
 (a) has at least about 20% of the biological activity of Ac-Lys-Pro-Ser-Ser-Pro-Pro-Glu-Glu-Am in one or more of the following in vitro bioassays: (i) invasion in a Matrigel® assay;  
 (ii) endothelial tube formation on Matrigel®, or (iii) endothelial tube formation on a fibrin matrix in the presence of basic fibroblast growth factor and vascular endothelial growth factor; or (b) competes with labeled Ac-Lys-Pro-Ser-Ser-Pro-Pro-Glu-Glu-Am for binding to a cell or molecule which has a binding site for Ac-Lys-Pro-Ser-Ser-Pro-Pro-Glu-Glu-Am.  
 
 
     
     
         2 . A peptide according to  claim 1  capped with an N-terminal acetyl group and a C terminal amide group.  
     
     
         3 . A substitution or addition variant of a peptide according to  claim 1 , or a chemical derivative of the variant, which variant has a sequence selected from the group consisting of: 
 (a) SEQ ID NO:2 wherein the Glu at position 7 or 8 or both is replaced by one or any two of the substituent amino acids Gln, Asp or Asn;    (b) SEQ ID NO:2 wherein Ser at position 3 or 4 or both is replaced by one or any two of the substituent amino acids Thr, Ala, Gly, hSer or ValβOH;    (c) SEQ ID NO:2 wherein the Lys at position 1 is replaced by His, Arg, Gln, Orn, Cit or Hci;    (d) SEQ ID NO:2 wherein the Pro at position 2, 5 or 6 is replaced by Hyp;    (e) an addition variant of SEQ ID NO:2, wherein Leu, Ile, Val, Nva, Nle, Met, Ala, or Gly is added to the C-terminal Glu or to any of said substituents for Glu at position 8;    (f) an addition variant of SEQ ID NO:2, wherein any of the following peptides are added to the C-terminal Glu or to any of said substituents for Glu at position 8: 
 Leu-(Gly) n ; Ile-(Gly) n ; Val-(Gly) n ; Nva-(Gly) n ; or Nle-(Gly) n , wherein n=1-10;  
   (g) an addition variant of SEQ ID NO:2 wherein one or more of the following residues or peptides is added to the N-terminal Lys or to any of said N-terminal substituents of Lys at position 1: 
 Gly, Lys-(Gly) n ; Tyr-(Gly) n ; or Gly-(Gly) n , wherein n=1-10; and  
   (h) a combination of one or more of (a)-(g).    
     
     
         4 . A compound according to  claim 1  which is a peptidomimetic agent.  
     
     
         5 . A multimer of a peptide or variant according to  claim 1  which, when the peptide is not a variant, has the formula: 
 (Lys-Pro-Ser-Ser-Pro-Pro-Glu-Glu-X m ) n -Lys-Pro-Ser-Ser-Pro-Pro-Glu-Glu wherein X is selected from the group consisting of C 1 -C 20  alkyl, C 1 -C 20  alkenyl, C 1 -C 20  alkynyl, C 1 -C 20  polyether containing up to 9 oxygen atoms and Gly z , and  
 wherein m=0 or 1, n=1-100 and z=1-10.  
 
     
     
         6 . A pharmaceutical composition useful for inhibiting (i) invasion of tumor cells or (ii) angiogenesis, comprising 
 (a) a peptide, variant or derivative according to  claim 1;  and    (b) a pharmaceutically acceptable carrier or excipient.    
     
     
         7 . A pharmaceutical composition useful for inhibiting (i) invasion of tumor cells or (ii) angiogenesis, comprising 
 (a) a peptide, variant or derivative according to  claim 2;  and    (b) a pharmaceutically acceptable carrier or excipient.    
     
     
         8 . A pharmaceutical composition useful for inhibiting (i) invasion of tumor cells or (ii) angiogenesis, comprising 
 (a) a peptide, variant or derivative according to  claim 3;  and    (b) a pharmaceutically acceptable carrier or excipient.    
     
     
         9 . A pharmaceutical composition useful for inhibiting (i) invasion of tumor cells or (ii) angiogenesis, comprising 
 (a) a peptidomimetic according to  claim 4;  and    (b) a pharmaceutically acceptable carrier or excipient.    
     
     
         10 . A pharmaceutical composition useful for inhibiting (i) invasion of tumor cells or (ii) angiogenesis, comprising 
 (a) a peptide multimer according to  claim 5;  and    (b) a pharmaceutically acceptable carrier or excipient.    
     
     
         11 . A method for inhibiting the invasiveness of tumor cells comprising contacting said cells with an effective amount of a peptide, variant or derivative according to  claim 1 .  
     
     
         12  A method for inhibiting cell migration, invasion, migration-induced cell proliferation or angiogenesis in a subject having a disease or condition associated with undesired cell migration, invasion, migration-induced proliferation, or angiogenesis, comprising administering to said subject an effective amount of a pharmaceutical composition according to  claim 6 .  
     
     
         13 . A method according to  claim 12  wherein said disease or condition is primary tumor growth, tumor invasion or metastasis, atherosclerosis, post-balloon angioplasty vascular restenosis, neointima formation following vascular trauma, vascular graft restenosis, fibrosis associated with a chronic inflammatory condition, lung fibrosis, chemotherapy-induced fibrosis, wound healing with scarring and fibrosis, psoriasis, deep venous thrombosis, or another disease or condition in which angiogenesis is pathogenic.  
     
     
         14 . A method according to  claim 13  wherein said disease is tumor growth, invasion or metastasis.