BMP Peptides & Methods of Use

Abstract

The invention relates to truncated growth factors and variants thereof. The invention also relates to methods of making and using the truncated growth factors. The invention further relates to compositions including a protease and a growth factor comprising a bone morphogenic protein (BMP) or a variant thereof. The invention also relates to methods of using the composition.

1 . A method of promoting differentiation of cells into osteoblasts, chondrocytes, ligament or tendon, the method comprising treating the cells with the composition of  claim 8  in an amount effective to promote the differentiation. 
     
     
         2 . The method of  claim 1 , wherein the cells are progenitor cells or adult stem cells. 
     
     
         3 . The method of  claim 2 , wherein the progenitor cells or the adult stem cells are derived from placenta, bone marrow, adipose tissue, blood vessel, amniotic fluid, synovial fluid, synovial membrane, pericardium, periosteum, dura, peripheral blood, umbilical blood, menstrual blood, teeth, nucleus pulposus, brain, skin, hair follicle, intestinal crypt, neural tissue, or muscle. 
     
     
         4 . The method of  claim 1 , wherein the cells are pluripotent stem cells. 
     
     
         5 . A method of promoting osteogenesis, chondrogenic, or ligament/tendon differentiation of cells in a tissue, the method comprising treating the tissue with the composition of  claim 8  in an amount effective to promote the osteogenesis, chondrogenic, or ligament/tendon differentiation. 
     
     
         6 . The method of  claim 5 , wherein the osteogenic activity of cells in the treated tissue is greater than the osteogenic activity of cells in untreated tissue. 
     
     
         7 . The method of  claim 5 , wherein the tissue is bone, cartilage, or connective tissue. 
     
     
         8 . A composition comprising
 a) a pharmaceutically acceptable carrier, and   b) a peptide of 137 residues or less, said peptide comprising an amino acid sequence at least 92% identical to the amino acid sequence of SEQ ID NO:20, 21, 22, 23, 24, 25, 26, 27 or 28, wherein the peptide has osteoinductive, chondroinductive, or ligament/tendon differentiating activity.   
     
     
         9 . The composition according to  claim 8 , wherein the peptide comprises the amino acid sequence of SEQ ID NO:20, 21, 22, 23, 24, 25, 26, 27 or 28. 
     
     
         10 . The composition according to  claim 9 , wherein the peptide consists of the amino acid sequence of SEQ ID NO:20, 21, 22, 23, 24, 25, 26, 27 or 28. 
     
     
         11 . A fusion protein comprising (a) a peptide of 137 residues or less, said peptide comprising an amino acid sequence at least 92% identical to the amino acid sequence of SEQ ID NO:20, 21, 22, 23, 24, 25, 26, 27 or 28, wherein the peptide has osteoinductive, chondroinductive, or ligament/tendon differentiating activity, and (b) a second peptide, wherein the second peptide comprises an amino acid sequence that is less than 70% identical to the amino acid sequence of SEQ ID NO:40, 41, 42, 43, 44, 45, 46, 47 or 48. 
     
     
         12 . A composition comprising
 a) a bone morphogenic protein (BMP), or an isolated peptide comprising an amino acid sequence at least 92% identical to any of amino acid sequences of SEQ ID NO: 16-35, wherein the peptide has osteoinductive, chondroinductive, or ligament/tendon differentiating activity, and   b) at least one protease that is not bone morphogenetic protein-1 (BMP-1).   
     
     
         13 . The composition according to  claim 12 , wherein the at least one protease is selected from the group consisting of collagenase, clostripain, dispase, trypsin, cathepsin, MMP-13 (matrix metalloproteinase-13), and a mixture thereof. 
     
     
         14 . A method of making the composition of  claim 8  comprising
 preparing said peptide by contacting mature BMP-5 with a protease under conditions that promote protein cleavage to produce the peptide and harvesting the peptide, and 
 mixing the peptide with the at least one pharmaceutically acceptable carrier. 
 
     
     
         15 . The method of  claim 14 , wherein the protease is selected from the group consisting of collagenase, clostripain, dispase, trypsin, cathepsin, BMP-1 (bone morphogenetic protein-1), MMP-13 (matrix metalloproteinase-13), and a mixture thereof. 
     
     
         16 - 17 . (canceled) 
     
     
         18 . A method of making the composition of  claim 8 , comprising
 preparing said peptide by culturing a host cell under conditions suitable for protein expression and harvesting the protein, wherein the host cell comprises a vector that encodes said peptide, and   mixing the peptide with the at least one pharmaceutically acceptable carrier.   
     
     
         19 . (canceled) 
     
     
         20 . A method of increasing a growth factor activity, the method comprising treating a cell with the composition of  claim 12  in an amount effective to increase the growth factor activity in the cell. 
     
     
         21 . The method according to  claim 20 , wherein the peptide is selected from the group consisting of
 a) a peptide of 137 residues or less, said peptide comprising an amino acid sequence at least 92% identical to the amino acid sequence of SEQ ID NO:20, 21, 22, 23, 24, 25, 26, 27 or 28,   b) a peptide of 137 residues or less, said peptide comprising the amino acid sequence of SEQ ID NO:20, 21, 22, 23, 24, 25, 26, 27 or 28, and   c) a peptide consisting of the amino acid sequence of SEQ ID NO:20, 21, 22, 23, 24, 25, 26, 27 or 28.   
     
     
         22 . The method according to  claim 20 , wherein the peptide is a fusion protein comprising (a) the peptide comprising the amino acid sequence of SEQ ID NO: 20, 21, 22, 23, 24, 25, 26, 27 or 28, and (b) a second peptide, wherein the second peptide comprises an amino acid sequence that is less than 70% identical to the amino acid sequence of SEQ ID NO: 45. 
     
     
         23 . The method according to  claim 20 , wherein the composition comprises two or more proteases. 
     
     
         24 . The method according to  claim 20 , wherein the at least one protease is selected from the group consisting of collagenase, clostripain, dispase, trypsin, cathepsin, MMP-13 (matrix metalloproteinase-13), and a mixture thereof. 
     
     
         25 . The method according to  claim 24 , wherein the at least one protease is collagenase, dispase or cathepsin. 
     
     
         26 . The method according to  claim 20 , wherein the cells are progenitor cells or adult stem cells. 
     
     
         27 . The method according to  claim 26 , wherein the progenitor cells or the adult stem cells are derived from placenta, bone marrow, adipose tissue, blood vessel, amniotic fluid, synovial fluid, synovial membrane, pericardium, periosteum, dura, peripheral blood, umbilical blood, menstrual blood, teeth, nucleus pulposus, brain, skin, hair follicle, intestinal crypt, neural tissue, or muscle. 
     
     
         28 . The method according to  claim 26 , wherein the progenitor cells or the adult stem cells are derived from pluripotent stem cells. 
     
     
         29 . The method according to  claim 20 , wherein the cells are pluripotent stem cells. 
     
     
         30 - 88 . (canceled) 
     
     
         89 . The composition according to  claim 8 , wherein the pharmaceutically acceptable carrier is selected from the group consisting of saline, Ringer's solution, dextrose solution, 5% human serum albumin, dispersion media, coatings, antibacterial and antifungal agents, and isotonic and absorption delaying agents. 
     
     
         90 . A method of growing and/or culturing cells comprising growing and/or culturing cells in the presence of the composition of  claim 8 . 
     
     
         91 . The method according to  claim 90 , wherein the cells are selected from the group consisting of mesenchymal stem cells, adipose-derived stem cells, embryonic stem cells, progenitor cells, differentiated cells, undifferentiated cells, and induced pluripotent stem cells.