Amphiphilic peptide matrices for treatment of osteoporosis

Abstract

The present invention provides methods for prevention, prevention of progression, and treatment of osteoporosis and pre-osteoporotic conditions comprising direct administering to osteoporotic bone of a composition comprising amphiphilic peptides and peptide matrices thereof, useful in promoting biomineralization, local osteoporotic medications and inducing bone repair.

What is claimed is: 
     
         1 . A method for treatment of progression of osteoporosis or pre-osteoporotic condition comprising promoting biomineralization to treat such condition in a patient in need thereof, by direct administration involving injection or implantation of a therapeutically effective amount of a composition comprising at least one amphiphilic peptide, derivative or salt thereof that forms a β-sheet structure that promotes biomineralization and an anti-resorptive agent, wherein said peptide comprises:
 i. 2-20 pairs of hydrophobic-hydrophilic alternating amino acid residues wherein the hydrophilic amino acid residue is selected from the group consisting of: a negatively charged amino acid, a hydroxyl-containing amino acid, and a phosphorylated-hydroxyl-containing amino acid; and 
 ii. no more than 10% positively charged amino acid residues. 
 
     
     
         2 . The method of  claim 1 , wherein the at least one amphiphilic peptide is 4-40 amino acids in length or 7-28 amino acids in length. 
     
     
         3 . The method of  claim 1 , wherein the at least one amphiphilic peptide comprises at least one terminal Pro residue or wherein at least one amphiphilic peptide comprises at least one moiety or amino acid residue capable of increasing the solubility of the peptide. 
     
     
         4 . The method of  claim 1 , wherein the hydrophobic amino acid residue is selected from the group consisting of Phe, Leu, Ile, Val or Ala, and wherein the hydrophilic amino acid residue is selected from the group consisting of: Glu, Asp, Tyr, Ser, Thr, Ser(PO 4 ), Thr(PO 4 ), and Tyr(PO 4 ). 
     
     
         5 . The method of  claim 1 , wherein the at least one amphiphilic peptide comprises an amino acid sequence according to Formula I:
   X-(hydrophobic-hydrophilic) n -B  (Formula I)
   wherein n designates an integer of 2-20, hydrophobic designates a hydrophobic amino acid residue, hydrophilic designates a hydrophilic amino acid residue, X designates Pro, Pro-hydrophilic or represents the peptide's amino terminus or a modified amino terminus, and B is Pro or represents the peptide's carboxy terminus or a modified carboxy terminus.   
     
     
         6 . The method of  claim 5 , wherein the at least one amphiphilic peptide comprises an amino acid sequence selected from the group consisting of:
 X-(Phe-Glu) n -B   X-(Phe-Asp) n -B   X-(Leu-Glu) n -B   X-(Leu-Asp) n -B   wherein n designates an integer of 2-20, X designates Pro, Pro-hydrophilic or represents the peptide's amino terminus or a modified amino terminus, hydrophilic designates a hydrophilic amino acid residue, and B is Pro or represents the peptide's carboxy terminus or a modified carboxy terminus.   
     
     
         7 . The method of  claim 5 , wherein the at least one amphiphilic peptide comprises a sequence selected from the group consisting of:
 Pro-Glu-(Phe-Glu) n  wherein n is an integer of 3-9;   Glu-(Phe-Glu) n -Pro wherein n is an integer of 3-9;   Pro-(Ser-Phe) n -Ser-Pro wherein n is an integer of 3-9;   Pro-(SerPO 4 -Phe) n -SerPO 4 -Pro wherein n is an integer of 3-9;   Pro-(TyrPO 4 -Phe) n -TyrPO 4 -Pro wherein n is an integer of 3-9;   Pro-(Glu-Leu) n -Glu-Pro wherein n is an integer of 3-9;   Pro-(Asp-Phe) n -Asp-Pro wherein n is an integer of 3-9;   Pro-(Asp-Leu) n -Asp-Pro wherein n is an integer of 3-9;   Pro-(Ser-Leu) 5 -Ser-Pro wherein n is an integer of 3-9;   Pro-(SerPO 4 -Leu) n -SerPO 4 -Pro wherein n is an integer of 3-9;   Pro-(TyrPO 4 -Leu) n -TyrPO 4 -Pro wherein n is an integer of 3-9;   Pro-(Glu-Phe-Ser-Phe) n -Glu-Pro wherein n is an integer of 3-9;   Pro-(SerPO 4 -Phe-Ser-Phe) n -Ser-Pro wherein n is an integer of 1-4;   Pro-(SerPO 4 -Phe-Glu-Phe) n -Glu-Pro wherein n is an integer of 1-4;   Pro-(SerPO 4 -Phe-Asp-Phe) n -Asp-Pro wherein n is an integer of 1-4;   Pro-Glu-(Phe-Glu) n -(Gly) m -Arg-Gly-Asp-Z wherein Z is Ser, Gly or represents the peptide's carboxy terminus, n is an integer of 2-15 and m is an integer of 0-10;   (Phe-Glu) n -(Gly) m -Arg-Gly-Asp-Z wherein Z is Ser, Gly or represents the peptide's carboxy terminus, n is an integer of 2-15 and m is an integer of 0-10;   Pro-(Asp-Phe) n -Asp-Pro-(Gly) m -Arg-Gly-Asp-Z wherein Z is Ser, Gly or represents the peptide's carboxy terminus, n is an integer of 2-15 and m is an integer of 0-10;   
       
         
           
                 
               
                   (SEQ ID NO: 1) 
                 
                   Pro-Glu-(Phe-Glu) 5 ; 
                 
                     
                 
                   (SEQ ID NO: 2) 
                 
                   Glu-(Phe-Glu) 5 -Pro; 
                 
                     
                 
                   (SEQ ID NO: 3) 
                 
                   Pro-(Ser-Phe) 5 -Ser-Pro; 
                 
                     
                 
                   (SEQ ID NO: 4) 
                 
                   Pro-(SerPO 4 -Phe) 5 -SerPO 4 -Pro; 
                 
                     
                 
                   (SEQ ID NO: 5) 
                 
                   Pro-(TyrPO 4 -Phe) 5 -TyrPO 4 -Pro; 
                 
                     
                 
                   (SEQ ID NO: 6) 
                 
                   Pro-(Glu-Leu) 5 -Glu-Pro; 
                 
                     
                 
                   (SEQ ID NO: 7) 
                 
                   Pro-(Asp-Leu) 5 -Asp-Pro; 
                 
                     
                 
                   (SEQ ID NO: 8) 
                 
                   Pro-(Ser-Leu) 5 -Ser-Pro; 
                 
                     
                 
                   (SEQ ID NO: 9) 
                 
                   Pro-(SerPO 4 -Leu) 5 -SerPO 4 -Pro; 
                 
                     
                 
                   (SEQ ID NO: 10) 
                 
                   Pro-(TyrPO 4 -Leu) 5 -TyrPO 4 -Pro; 
                 
                     
                 
                   (SEQ ID NO: 11) 
                 
                   Pro-(Glu-Phe-Ser-Phe) 4 -Glu-Pro; 
                 
                     
                 
                   (SEQ ID NO: 12) 
                 
                   Pro-(SerPO 4 -Phe-Ser-Phe) 4 -Ser-Pro; 
                 
                     
                 
                   (SEQ ID NO: 13) 
                 
                   Pro-(SerPO 4 -Phe-Glu-Phe) 4 -Glu-Pro; 
                 
                     
                 
                   (SEQ ID NO: 14) 
                 
                   Pro-(SerPO 4 -Phe-Asp-Phe) 4 -Asp-Pro; 
                 
                     
                 
                   (SEQ ID NO: 15) 
                 
                   Ala-Leu-Glu-(Phe-Glu) 3 -Pro-Ala-(Glu-Phe) 3 -Glu-Leu- 
                 
                     
                 
                   Pro-Ala-Leu-Glu-(Phe-Glu) 3 -Pro; 
                 
                   (SEQ ID NO: 16) 
                 
                   Pro-Glu-(Phe-Glu) 2 -Lys-(Glu-Phe) 2 -Glu-Pro; 
                 
                     
                 
                   (SEQ ID NO: 17) 
                 
                   Pro-Glu-(Phe-Glu) 5 -(Gly) 3 -Arg-Gly-Asp-Ser; 
                 
                     
                 
                   (SEQ ID NO: 18) 
                 
                   (Phe-Glu) 3 -Pro-(Gly) 3 -Arg-Gly-Asp-Ser; 
                 
                     
                 
                   (SEQ ID NO: 19) 
                 
                   Ac-Pro-Asp-(Phe-Asp) 5 -Pro-NH 2 ;; 
                 
                     
                 
                   (SEQ ID NO: 20) 
                 
                   Pro-Asp-(Phe-Asp) 6 ; 
                 
                     
                 
                   (SEQ ID NO: 21) 
                 
                   (Phe-Asp) 6 ; 
                 
                     
                 
                   (SEQ ID NO: 22) 
                 
                   Pro-Glu-(Phe-Glu) 5 -Pro; 
                 
                     
                 
                   (SEQ ID NO: 23) 
                 
                   Pro-Asp-(Phe-Asp) 5 -Pro-NH 2 ;; 
                 
                     
                 
                   (SEQ ID NO: 24) 
                 
                   (Phe-Glu) 5 ; 
                 
                     
                 
                   (SEQ ID NO: 25) 
                 
                   (Phe-Glu) 6 ; 
                 
                     
                 
                   (SEQ ID NO: 26) 
                 
                   (Phe-Glu) 7 ; 
                 
                     
                 
                   (SEQ ID NO: 27) 
                 
                   Pro-Asp-(Phe-Asp) 4 ; 
                 
                     
                 
                   (SEQ ID NO: 28) 
                 
                   Pro-Asp-(Phe-Asp) 6 ; 
                 
                     
                 
                   (SEQ ID NO: 29) 
                 
                   Pro-Asp-(Phe-Asp) 8 ; 
                 
                     
                 
                   (SEQ ID NO: 30) 
                 
                   (Phe-Asp) 5 ; 
                 
                     
                 
                   (SEQ ID NO: 31) 
                 
                   (Phe-Asp) 6 ; 
                 
                     
                 
                   (SEQ ID NO: 32) 
                 
                   (Phe-Asp) 7 ; 
                 
                     
                 
                   (SEQ ID NO: 33) 
                 
                   Pro-Asp-(Phe-Asp) 5 -Pro-Arg-Gly-Asp-Ser; 
                 
                     
                 
                   (SEQ ID NO: 34) 
                 
                   Pro-(Phe-Asp) 3 -Pro; 
                 
                   and 
                 
                     
                 
                   SEQ ID NO: 35) 
                 
                   Pro-(Phe-Asp) 3 -Pro-(Gly) 3 -Arg-Gly-Asp-Ser,. 
                 
             
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
               
            
           
         
       
     
     
         8 . The method of  claim 1 , wherein the anti-resorptive agent is selected from the group consisting of: bisphosphonates, estrogen inhibitors, parathyroid hormone (PTH), fibroblast growth factor (FGF), insulin growth factor (IGF), and calcitonin. 
     
     
         9 . The method of  claim 8 , wherein the anti-resorptive agent is a bisphosphonate selected from the group consisting of: alendronate, clodronate, etidronate, ibandronate, icadronate, pamidronate, risedronate, tiludronate and zoledronate. 
     
     
         10 . The method of  claim 1 , wherein the composition further comprises pre-loaded mineral-salt solution or aggregates or wherein the composition comprises pre-loaded polysaccharide. 
     
     
         11 . The method of  claim 10 , wherein the pre-loaded mineral salt is a calcium phosphate mineral selected from the group consisting of amorphous calcium phosphate, tricalcium phosphate or hydroxyapatite or wherein the pre-loaded polysaccharide is selected from the group consisting of: hyaluronic acid, alginate and sulfated polysaccharide. 
     
     
         12 . The method of  claim 10 , wherein the composition further comprises the sequence Ac-Pro-Asp-(Phe-Asp) 5 -Pro-NH 2  and alginate. 
     
     
         13 . The method of  claim 1 , wherein the compositing further comprises at least one other therapeutic agent. 
     
     
         14 . The method of  claim 13 , wherein the therapeutic agent is selected from the group consisting of: active protein, growth factor, cytokine, chemotherapeutic drug, enzyme, anti-microbial, anti-resorptive agent, hormone, and anti-inflammatory agent. 
     
     
         15 . The method of  claim 1 , wherein the composition is a dry composition or a semi-fluid composition or wherein the composition is selected from the group consisting of particles, granules, paste, dough, powder, gel or hydrogel. 
     
     
         16 . The method of  claim 1 , wherein the composition is administered into the hip, into a vertebral bone in the spine, into the interior of a vertebra or into a medullary canal of a long bone or wherein the composition is administered into a bone area selected from the group consisting of compact bone, cancellous bone, epiphyseal line, epiphysis, and metaphysic. 
     
     
         17 . The method of  claim 1 , wherein the composition is administered directly into deficient, deteriorated, and injured bone and low bone mineral density sites.