Concentrated protein preparations of bone morphogenetic proteins and methods of use thereof

Abstract

Disclosed herein are heretofore undescribed preparations of highly concentrated, solubilized proteins, such as but not limited to, Bone Morphogenetic Proteins. Such protein preparations can be formulated in an aqueous carrier at protein concentrations in excess of 10 mg/ml when using the methods of manufacture taught herein. Such methods yield stable protein preparations in either solubilized or lyophilized form. The protein preparations of the present invention are particularly beneficial when administered either locally or systemically, in part, because low administration volumes can be accomplished. This is especially important for local treatment of certain anatomic locations such as, for example, the synovial fluid of a joint when treating osteoarthritis with BMP-7 or the intradiscal space when treating degenerative disc disease with BMP-7.

We claim:  
     
         1 . A preparation of bone morphogenetic protein comprising:
 an aqueous carrier; and, bone morphogenetic protein solubilized in said carrier at a concentration of at least about 10 mg/ml.   
     
     
         2 - 29 . (canceled) 
     
     
         30 . A method of treating a skeletal tissue disorder, injury or disease, the method comprising the step of:
 administering to a subject in need thereof the protein preparation of  claim 1 , wherein said protein preparation is in a dose effective to treat said skeletal tissue disorder, injury or disease.   
     
     
         31 - 45 . (canceled) 
     
     
         46 . A method of treating a non-skeletal tissue, the method comprising the step of; administering to a subject in need thereof the protein preparation of  claim 1 , wherein said protein preparation is in a dose effective to treat said non-skeletal tissue disorder, injury or disease. 
     
     
         47 . (canceled) 
     
     
         48 . A kit 1) comprising a lyophilized bone morphogenetic protein and a reconstitution diluent, wherein the protein and the diluent are in separate containers, and wherein the amount of diluent is sufficient only for preparation of solubilized protein at a concentration of at least about 10 mg/ml; or 2) for the treatment of a disorder, injury or disease comprising a lyophilized bone morphogenetic protein and a reconstitution diluent, wherein the protein and the diluent are in separate containers, and further wherein the kit comprises a plurality of separate containers each containing an amount of diluent, such that the amounts of diluent are sufficient to prepare solubilized protein preparations ranging in concentration from at least about 2 mg/ml to about 60 mg/ml for use in the treatment of a disorder, injury or disease. 
     
     
         49 - 50 . (canceled) 
     
     
         51 . A method A) for manufacturing a concentrated form of lyophilized recombinant bone morphogenetic protein, the method comprising the steps of:
 (1) providing a processing solution of recombinant bone morphogenetic protein, the weight per volume of protein in said processing solution being less than the weight per volume specified for lyophilization;   (2) adjusting the weight per volume of protein in said processing solution to produce a lyophilization solution, the weight per volume of protein in said lyophilization solution being greater than that of the processing solution;   (3) filling a vial suitable for lyophilization with a specified volume of said lyophilization solution; and,   (4) lyophilizing said lyophilization solution to manufacture a concentrated form of lyophilized recombinant bone morphogenetic protein;   B) for preparing a concentrated form of bone morphogenetic protein, the method comprising the steps of:   (1) providing a processing solution of bone morphogenetic protein, the weight per volume of protein in said processing solution being the same as the weight per volume specified for lyophilization;   (2) filling a vial suitable for lyophilization with a specified volume of said processing solution thereby producing a lyophilization solution;   (3) lyophilizing said lyophilization solution; and,   (4) packaging the lyophilized lyophilization solution together with a specified volume of reconstitution diluent, the volume of said diluent being less than the specified fill volume of step 3; or   C) for preparing a concentrated form of protein in lyophilized or other reconstitutible non-liquid form, the method comprising the steps of:   (1) providing a processing solution of protein, the weight per volume of protein in said processing solution being the same as the weight per volume specified for lyophilization or other reconsitutible non-liquid form;   (2) providing a vial suitable for lyophilization (or other form) for containing a specified volume of said processing solution thereby producing a lyophilization (or other form) solution;   wherein the weight per volume of protein in each of the processing and lyophilization (or other form) solution is at least 2 mg/ml.   
     
     
         52 - 54 . (canceled) 
     
     
         55 . A concentrated form of recombinant bone morphogenetic protein according to  claim 51 . 
     
     
         56 - 62 . (canceled)