Freeze-dried polymer compositions for mixing with platelet rich plasma to form implants for tissue repair and/or compositions for therapeutic intra-articular injections

Abstract

The present application relates to a freeze-dried polymer composition containing chitosan and at least one lyoprotectant, a process for preparing a freeze-dried composition containing chitosan and at least one lyoprotectant and the use of a reconstituted freeze-dried chitosan composition to prepare implants for tissue repair.

1 . A freeze-dried homogeneous solid cake comprising chitosan having a molecular weight number of from about 4 kDa to about 250 kDa and a degree of deacetylation of from about 80% to about 85%, a clot activating amount of a clot activator, and at least one lyoprotectant, wherein said composition is without glycerol phosphate and said freeze-dried homogeneous solid cake is soluble in blood or a blood product selected from the group consisting of platelet rich plasma (PRP), platelet-poor plasma (PPP), autologous conditioned plasma, platelet suspension, platelet lysate and combinations thereof for reconstitution of an injectable solution forming an injectable solution which upon injection:
 i) into tissue solidifies forming an implant for tissue repair; or   ii) into an articular joint mixes with intra-articular fluids.   
     
     
         2 . The freeze-dried homogeneous solid cake of  claim 1  wherein the at least one lyoprotectant is selected from the group consisting of monosaccharide, polyol, disaccharide, trisaccharide, oligosaccharide/polysaccharide, high molecular weight excipient, amino acid, protein and a combination thereof. 
     
     
         3 . The freeze-dried homogeneous solid cake of  claim 1  wherein the chitosan has a molecular weight number from about 20 to about 250 kDa. 
     
     
         4 . The freeze-dried homogeneous solid cake of  claim 1  wherein the chitosan is present from about 0.25% to about 10% w/v. 
     
     
         5 . The freeze-dried homogeneous solid cake of  claim 1  wherein the at least one lyoprotectant is present from about 0.1% to about 30% w/v. 
     
     
         6 . The freeze-dried homogeneous solid cake of  claim 1  wherein the clot activator is selected from the group consisting of calcium chloride, calcium gluconate, calcium acetate, calcium carbonate, calcium glubionate, calcium gluceptate, calcium lactate, calcium lactobionate, calcium phosphate and combinations thereof. 
     
     
         7 . A process for preparing a freeze-dried homogeneous solid cake comprising the steps of:
 a) contacting chitosan with water to form a chitosan/water aqueous mixture,   b) contacting the aqueous mixture with at least one lyoprotectant,   c) contacting the aqueous mixture with a clot activating amount of at least one clot activator forming a chitosan/water/clot activator mixture,   d) sterilizing the chitosan, the at least one lyoprotectant and the at least one clot activator, individually, prior to mixing, or after addition of the at least one lyoprotectant and the at least one clot activator to said chitosan/water aqueous mixture; and   e) freeze-drying the chitosan/water/clot activator mixture.   
     
     
         8 . The process of  claim 7  wherein the chitosan has a molecular weight number from about 4 to about 250 kDa. 
     
     
         9 . The process of  claim 7  wherein the chitosan is present from about 0.25% to about 10% w/v. 
     
     
         10 . The process of  claim 7  wherein the at least one lyoprotectant is selected from the group consisting of monosaccharide, polyol, disaccharide, trisaccharide, oligosaccharide/polysaccharide, high molecular weight excipient, amino acid, protein and a combination thereof. 
     
     
         11 . The process of  claim 7  wherein the at least one lyoprotectant is present from about 0.1% to about 30% w/v. 
     
     
         12 . A freeze-dried homogeneous solid cake consisting of: chitosan at a concentration of no more than about 1% (w/v), having a molecular weight in the range of about M n  28 to about 56 kDa and at least one lyoprotectant at a concentration of no more than about 4% (w/v), wherein said freeze-dried homogeneous solid cake is soluble in blood or a blood product selected from the group consisting of platelet rich plasma (PRP), platelet-poor plasma (PPP), autologous conditioned plasma, platelet suspension, platelet lysate and combinations thereof for reconstitution of an injectable solution, wherein said freeze-dried homogenous solid cake further comprises a clot activating amount of a clot activator. 
     
     
         13 . A freeze-dried homogeneous solid cake consisting of: chitosan at a concentration of no more than about 0.56% (w/v), having a molecular weight in the range of about M n  89 to about 108 kDa and at least one lyoprotectant at a concentration of no more than about 4% (w/v), wherein said freeze-dried homogeneous solid cake is soluble in blood or a blood product selected from the group consisting of platelet rich plasma (PRP), platelet-poor plasma (PPP), autologous conditioned plasma, platelet suspension, platelet lysate and combinations thereof for reconstitution of an injectable solution, wherein said freeze-dried homogenous solid cake further comprises a clot activating amount of a clot activator. 
     
     
         14 . A freeze-dried homogeneous solid cake comprising: chitosan at a concentration range of about 0.42 and about 2% (w/v), having a molecular weight range of about M n  4 kDa to about 211 kDa and a degree of deacetylation of from about 80% to about 85%, at least one lyoprotectant at a concentration range between about 1 and about 10%, and a salt or a buffer, and being without glycerol phosphate, wherein said freeze-dried homogeneous solid cake is soluble in blood or a blood product selected from the group consisting of platelet rich plasma (PRP), platelet-poor plasma (PPP), autologous conditioned plasma, platelet suspension, platelet lysate and combinations thereof for reconstitution of an injectable solution, wherein said salt is a clot activator present in a clot activating amount. 
     
     
         15 . A freeze-dried homogeneous solid cake comprising chitosan having a molecular weight number of from about 4 kDa to about 250 kDa and a degree of deacetylation of from about 80% to about 85% and at least one lyoprotectant, wherein said freeze-dried homogeneous solid cake is soluble in blood or a blood product selected from the group consisting of platelet rich plasma (PRP), platelet-poor plasma (PPP), autologous conditioned plasma, platelet suspension, platelet lysate and combinations thereof for reconstitution of an injectable solution forming an injectable solution, exhibiting at least one of the following:
 in situ coagulation achieved and not inhibited;   able to withstand mechanical loading post-implantation;   inhibition of platelet-mediated clot retraction to fill tissue defects;   good mixing without phase separation of polymers and PRP, blood product and combinations thereof;   viscous and paste-like formulations for tissue repair applications;   close-to-physiological properties for in vivo application; and combinations thereof, wherein said freeze-dried homogenous solid cake further comprises a clot activating amount of a clot activator.   
     
     
         16 . The freeze-dried homogeneous solid cake of  claim 12  wherein said composition is without glycerol phosphate. 
     
     
         17 . The freeze-dried homogeneous solid cake of  claim 13  wherein said composition is without glycerol phosphate. 
     
     
         18 . A freeze-dried homogeneous solid cake consisting of: chitosan at a concentration range of about 0.42 and about 2% (w/v), having a molecular weight range of about M n  4 to about 211 kDa and a degree of deacetylation of from about 80% to about 85%, at least one lyoprotectant at a concentration range between about 1 and about 10%, and a salt or a buffer, and being without glycerol phosphate, wherein said freeze-dried homogeneous solid cake is soluble in blood or a blood product selected from the group consisting of platelet rich plasma (PRP), platelet-poor plasma (PPP), autologous conditioned plasma, platelet suspension, platelet lysate and combinations thereof for reconstitution of an injectable solution, wherein said salt is a clot activator present in a clot activating amount. 
     
     
         19 . A freeze-dried homogeneous solid cake consisting of chitosan having a molecular weight number of from about 4 kDa to about 250 kDa and a degree of deacetylation of from about 80% to about 85%, a clot activating amount of a clot activator and at least one lyoprotectant, wherein said freeze-dried homogeneous solid cake is soluble in blood or a blood product selected from the group consisting of platelet rich plasma (PRP), platelet-poor plasma (PPP), autologous conditioned plasma, platelet suspension, platelet lysate and combinations thereof for reconstitution of an injectable solution forming an injectable solution, exhibiting at least one of the following:
 in situ coagulation achieved and not inhibited;   able to withstand mechanical loading post-implantation;   inhibition of platelet-mediated clot retraction to fill tissue defects;   good mixing without phase separation of polymers and PRP, blood product and combinations thereof;   viscous and paste-like formulations for tissue repair applications;   close-to-physiological properties for in vivo application; and combinations thereof.   
     
     
         20 . The freeze-dried homogenous solid cake of  claim 1  wherein said freeze-dried homogenous solid cake is soluble in said blood or blood product in 5 minutes or less. 
     
     
         21 . The freeze-dried homogeneous solid cake of  claim 12  wherein said freeze-dried homogenous solid cake is soluble in said blood or blood product in 5 minutes or less. 
     
     
         22 . A freeze-dried homogeneous solid cake consisting of: chitosan at a concentration of no more than about 1% (w/v), having a molecular weight in the range of about M n  28 to about 56 kDa, at least one lyoprotectant at a concentration of no more than about 4% (w/v), and a clot activating amount of a clot activator, wherein said freeze-dried homogeneous solid cake is soluble in blood or a blood product selected from the group consisting of platelet rich plasma (PRP), platelet-poor plasma (PPP), autologous conditioned plasma, platelet suspension, platelet lysate and combinations thereof for reconstitution of an injectable solution. 
     
     
         23 . The freeze-dried homogeneous solid cake of  claim 13  wherein said freeze-dried homogenous solid cake is soluble in said blood or blood product in 5 minutes or less. 
     
     
         24 . A freeze-dried homogeneous solid cake consisting of: chitosan at a concentration of no more than about 0.56% (w/v), having a molecular weight in the range of about M n  89 kDa to about 108 kDa and at least one lyoprotectant at a concentration of no more than about 4% (w/v), a clot activating amount of a clot activator, wherein said freeze-dried homogeneous solid cake is soluble in blood or a blood product selected from the group consisting of platelet rich plasma (PRP), platelet-poor plasma (PPP), autologous conditioned plasma, platelet suspension, platelet lysate and combinations thereof for reconstitution of an injectable solution. 
     
     
         25 . The freeze-dried homogeneous solid cake of  claim 14  wherein said freeze-dried homogenous solid cake is soluble in said blood or blood product in 5 minutes or less. 
     
     
         26 . The freeze-dried homogeneous solid cake of  claim 15  wherein said freeze-dried homogenous solid cake is soluble in said blood or blood product in 5 minutes or less. 
     
     
         27 . The freeze-dried homogeneous solid cake of  claim 18  wherein said freeze-dried homogenous solid cake is soluble in said blood or blood product in 5 minutes or less. 
     
     
         28 . A freeze-dried homogeneous solid cake consisting of: chitosan at a concentration range of about 0.42 and about 2% (w/v), having a molecular weight range of about M n  4 kDa to about 211 kDa and a degree of deacetylation of from about 80% to about 85%, at least one lyoprotectant at a concentration range between about 1 and about 10%, and a salt or a buffer, and being without glycerol phosphate, wherein said freeze-dried homogeneous solid cake is soluble in blood or a blood product selected from the group consisting of platelet rich plasma (PRP), platelet-poor plasma (PPP), autologous conditioned plasma, platelet suspension, platelet lysate and combinations thereof for reconstitution of an injectable solution, wherein said salt is a clot activating amount of a clot activator. 
     
     
         29 . The freeze-dried homogeneous solid cake of  claim 19  wherein said freeze-dried homogenous solid cake is soluble in said blood or blood product in 5 minutes or less. 
     
     
         30 . A freeze-dried homogeneous solid cake consisting of chitosan having a molecular weight number of from about 4 kDa to about 250 kDa and a degree of deacetylation of from about 80% to about 85% and at least one lyoprotectant, a clot activating amount of a clot activator, wherein said freeze-dried homogeneous solid cake is soluble in blood or a blood product selected from the group consisting of platelet rich plasma (PRP), platelet-poor plasma (PPP), autologous conditioned plasma, platelet suspension, platelet lysate and combinations thereof for reconstitution of an injectable solution forming an injectable solution, exhibiting at least one of the following:
 in situ coagulation achieved and not inhibited;   able to withstand mechanical loading post-implantation;   inhibition of platelet-mediated clot retraction to fill tissue defects;   good mixing without phase separation of polymers and PRP, blood product and combinations thereof;   viscous and paste-like formulations for tissue repair applications;   close-to-physiological properties for in vivo application; and combinations thereof.