Agents, Which Inhibit Apoptosis in Cells that are Involved in Wound Healing

Abstract

The invention relates to the use of substances as a fundamental constituent in wound healing agents. The invention is characterized in that said substances bond to either IAP and/or integrin α v β 3 and/or thrombospondin-1 in such a way that the bond between thrombospondin-1 and IAP and/or integrin α v β 3 is inhibited.

1 - 26 . (canceled) 
     
     
         27 . A method for promoting wound healing, the method comprising administering to a subject in need of such treatment an effective amount of a composition comprising
 a substance that binds to at least one of integrin-associated protein (IAP), integrin α v β 3 , and thrombospondin 1, wherein said substance inhibits binding between thrombospondin 1 and at least one of IAP and integrin α v β 3  and inhibits apoptosis characterized by nuclear fragmentation of cells, wherein said substance is an antibody or functional variant thereof that specifically binds at least one of thrombospondin 1, IAP and an integrin.   
     
     
         28 . The method of  claim 27 , wherein the substance binds to at least one of IAP and integrin α v β 3  on fibroblasts and/or to thrombospondin 1 in such a way that the binding between thrombospondin 1 and the at least one of IAP and integrin α v β 3  is inhibited and the rate of apoptosis of the cells is reduced by more than 10%. 
     
     
         29 . The method of  claim 28 , wherein the cells are epithelial cells. 
     
     
         30 . The method of  claim 28 , wherein the cells are keratinocytes. 
     
     
         31 . The method of  claim 27 , wherein the substance is prepared by carrying out an identification method comprising steps (i) to (v), as follows:
 (i) culturing cells which express both IAP and integrin α v β 3 ,   (ii) causing the cells to produce an apoptosis-inducing material, and/or adding a material or materials inducing apoptosis,   (iii) adding a substance,   (iv) measuring the rate of apoptosis, and   (v) selecting identificate substances which cause a reduced rate of apoptosis, and then   mixing the identificates with a pharmaceutically acceptable carrier.   
     
     
         32 . The method of  claim 31 , wherein the cells employed in the identification method are endothelial cells. 
     
     
         33 . The method of  claim 31 , wherein the cells employed in the identification method are smooth muscle cells or fibroblasts. 
     
     
         34 . The method of  claim 31 , wherein the cells employed in the identification method are genetically modified cells which express both IAP and α v β 3  on their surface. 
     
     
         35 . The method of  claim 31 , wherein the cells employed in the identification method are cultivated under conditions under which consistently directed laminar flows do not occur. 
     
     
         36 . The method of  claim 31 , wherein thrombospondin 1 or an analogous compound with identical binding properties is added to the cell culture in the identification method. 
     
     
         37 . The method of  claim 31 , wherein the induction of apoptosis in the identification method is determined by measuring increased calcium influx into the cell. 
     
     
         38 . The method of  claim 31 , wherein the rate of apoptosis in the identification method is determined by a method selected from the group consisting of DAPI staining, TUNEL assay, DNA ladder, annexin staining and enzyme detection. 
     
     
         39 . The method of  claim 31 , wherein the identificates bind to a calcium channel in the cell membrane in such a way that apoptosis-specific calcium influx into the cells is suppressed. 
     
     
         40 . The method of  claim 31 , wherein the identificates inhibit apoptosis-specific calcium influx into fibroblasts. 
     
     
         41 . The method of  claim 31 , wherein the process comprises admixing at least one different fibroblast growth factor to the medicament in addition to the identificates. 
     
     
         42 . The method of  claim 27 , further comprising at least one fibroblast growth factor. 
     
     
         43 . The method of  claim 42 , wherein the fibroblast growth factor is basic fibroblast growth factor. 
     
     
         44 . A medicament for promoting wound healing comprising a substance that binds to at least one of integrin-associated protein (IAP), integrin α v β 3 , and thrombospondin 1, wherein said substance inhibits binding between thrombospondin 1 and at least one of IAP and integrin α v β 3 , and inhibits apoptosis characterized by nuclear fragmentation of cells, wherein said substance is an antibody or functional variant thereof that specifically binds at least one of thrombospondin 1, IAP and an integrin. 
     
     
         45 . The medicament of  claim 44 , wherein the substance binds to at least one of IAP and integrin α v β 3  on fibroblasts and/or to thrombospondin 1 in such a way that the binding between thrombospondin 1 and the at least one of IAP and integrin α v β 3  is inhibited and the rate of apoptosis of the cells is reduced by more than 10%. 
     
     
         46 . The medicament of  claim 45 , wherein the cells are epithelial cells. 
     
     
         47 . The medicament of  claim 45 , wherein the cells are keratinocytes. 
     
     
         48 . The medicament of  claim 44  prepared by carrying out an identification method comprising steps (i) to (v), as follows:
 (i) culturing cells which express both IAP and integrin α v β 3      (ii) causing the cells to produce an apoptosis-inducing material, and/or adding a material or materials inducing apoptosis,   (iii) adding a substance,   (iv) measuring the rate of apoptosis, and   (v) selecting identificate substances which cause a reduced rate of apoptosis, and then   mixing the identificates with a pharmaceutically acceptable carrier.   
     
     
         49 . The medicament of  claim 48 , wherein the cells employed in the identification method are endothelial cells. 
     
     
         50 . The medicament of  claim 48 , wherein the cells employed in the identification method are smooth muscle cells or fibroblasts. 
     
     
         51 . The medicament of  claim 48 , wherein the cells employed in the identification method are genetically modified cells which express both IAP and α v β 3  on their surface. 
     
     
         52 . The medicament of  claim 48 , wherein the cells employed in the identification method are cultivated under conditions under which consistently directed laminar flows do not occur. 
     
     
         53 . The medicament of  claim 48 , wherein thrombospondin 1 or an analogous compound with identical binding properties is added to the cell culture in the identification method. 
     
     
         54 . The medicament of  claim 48 , wherein the induction of apoptosis in the identification method is determined by measuring increased calcium influx into the cell. 
     
     
         55 . The medicament of  claim 48 , wherein the rate of apoptosis in the identification method is determined by a method selected from the group consisting of DAPI staining, TUNEL assay, DNA ladder, annexin staining and enzyme detection. 
     
     
         56 . The medicament of  claim 48 , wherein the identificates bind to a calcium channel in the cell membrane in such a way that apoptosis-specific calcium influx into the cells is suppressed. 
     
     
         57 . The medicament of  claim 48 , wherein the identificates inhibit apoptosis-specific calcium influx into fibroblasts. 
     
     
         58 . The medicament of  claim 48 , wherein the process comprises admixing at least one different fibroblast growth factor to the medicament in addition to the identificates. 
     
     
         59 . The medicament of  claim 44 , further comprising at least one fibroblast growth factor. 
     
     
         60 . The medicament of  claim 59 , wherein the fibroblast growth factor is basic fibroblast growth factor. 
     
     
         61 . The medicament of  claim 44 , further comprising a pharmaceutical acceptable carrier.