Bone Morphogenetic Protein Compositions

Abstract

The present invention relates to compositions of bone morphogenetic proteins, particularly solid and liquid formulations of such proteins comprising one or more stabilizing excipients. The invention further provides methods of producing the compositions, kits comprising the compositions and methods of using the compositions in the treatment of diseases of skeletal and non-skeletal tissues.

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 . The protein preparation of  claim 1 , further comprising a stabilizing excipient. 
   
   
       3 . The protein preparation of  claim 2 , wherein the stabilizing excipient is selected from the group consisting of: sugars, polyols, surfactants, and any combination thereof. 
   
   
       4 . The protein preparation of  claim 3 , wherein the stabilizing excipient is selected from the group consisting of glucose, sucrose, raffinose, trehalose, lactose, mannitol, sorbitol, Tween 80, Tween 20 and Pluronic F-68 and combinations thereof. 
   
   
       5 . The protein preparation of  claim 4 , wherein the stabilizing excipient is trehalose. 
   
   
       6 . The protein preparation of  claim 5  having a pH of from about 2.5 to about 3.5. 
   
   
       7 . The protein preparation of  claim 5 , wherein the aqueous carrier comprises a buffer of the single acidic group type selected from the group consisting of: potassium phosphate, proprionic acid, lactic acid, trifluoroacetic acid and acetic acid; or the two acidic group type selected from the group consisting of: sodium glutamate and sodium succinate. 
   
   
       8 . The protein preparation of  claim 5 , wherein the protein is selected from the group consisting of BMP-2, BMP-4, BMP-5, BMP-6, BMP-7, GDF-5, GDF-6 GDF-7, and sequence variants of any one of the foregoing. 
   
   
       9 . The protein preparation of  claim 8 , wherein the protein is selected from the group consisting of GDF-5, GDF-6 and GDF-7. 
   
   
       10 . The protein preparation of  claim 8 , wherein the protein is selected from the group consisting of BMP-7. 
   
   
       11 . 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 5 , wherein said protein preparation is in a dose effective to treat said skeletal tissue disorder, injury or disease. 
   
   
       12 . The method of  claim 11 , wherein the skeletal tissue is mineralized or non-mineralized skeletal tissue. 
   
   
       13 . The method of  claim 12 , wherein the protein preparation is in an amount effective to treat a skeletal tissue disorder, injury or disease selected from the group consisting of metabolic bone disease, osteoarthritis, osteochondral disease, rheumatoid arthritis, osteoporosis, bone fractures, Paget's disease, periodontitis, and dentinogenesis. 
   
   
       14 . The method of  claim 12 , wherein the protein preparation is in an amount effective to treat a non-mineralized skeletal tissue disorder, injury or disease selected from the group consisting of osteoarthritis, osteochondral disease or defect, chondral disease or defect, rheumatoid arthritis, trauma-induced and inflammation-induced cartilage degeneration, age-related cartilage degeneration, articular cartilage injuries and diseases, full thickness cartilage defects, superficial cartilage defects, sequelae of systemic lupus erythematosis, sequelae of scleroderma, periodontal tissue regeneration, hierniation and rupture of intervertebral discs, degenerative diseases of the intervertebral disc, osteocondrosis, and injuries and diseases of ligament, tendon, synovial capsule, synovial membrane and meniscal tissues. 
   
   
       15 . The method of  claim 11 , wherein the protein preparation is in an amount effective to ameliorate tissue injury selected from the group consisting of: trauma-induced and inflammation-induced cartilage degeneration, articular cartilage injuries, full thickness cartilage defects, superficial cartilage defects, hierniation and rupture of intervertebral discs, degeneration of intervertebral discs due to an injury(s), and injuries of ligament, tendon, synovial capsule, synovial membrane and meniscal tissues. 
   
   
       16 . The method of  claim 11 , wherein the disease is degenerative disc disease. 
   
   
       17 . The method of  claim 16 , wherein the effective dose is administered to the intradiscal space, the nucleus pulposus or the annulus fibrosus. 
   
   
       18 . The method of  claim 11 , wherein the method further comprises the step of admixing said protein preparation with a suitable matrix material prior to administration. 
   
   
       19 . The method of  claim 11 , wherein the protein is selected from the group consisting of: BMP-2, BMP-4, BMP-5, BMP-6, BMP-7, GDF-5, GDF-6 GDF-7, and sequence variants of any one of the foregoing. 
   
   
       20 . The method of  claim 19 , wherein the protein is BMP-7. 
   
   
       21 . A kit comprising a lyophilized bone morphogenetic protein preparation 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. 
   
   
       22 . The kit of  claim 21 , further comprising a matrix. 
   
   
       23 . The kit of  claim 21 , further comprising an implantable device suitable for coating with the solubilized protein. 
   
   
       24 . The kit of  claim 21 , wherein the lyophilized bone morphogenetic protein preparation comprises a stabilizing excipient. 
   
   
       25 . The kit of  claim 24 , wherein the stabilizing excipient is trehalose. 
   
   
       26 . A composition comprising at least one BMP and an amount of trehalose sufficient to stabilize said BMP. 
   
   
       27 . The composition of  claim 26 , wherein said BMP is rhGDF-5. 
   
   
       28 . The composition of  claim 26 , further comprising at least one excipient selected from the group consisting of glycine, polysorbate 80, polysorbate 20, and mixtures thereof. 
   
   
       29 . The composition of  claim 28 , having a pH of about 2.5 to about 3.5. 
   
   
       30 . A method of stabilizing a BMP comprising the steps of: a.) providing a composition containing at least one BMP and an amount of trehalose sufficient to stabilize said BMP, and b.) lyophilizing the composition. 
   
   
       31 . The method of  claim 30 , wherein said BMP is rhGDF-5. 
   
   
       32 . The method of  claim 30 , further comprising the step of adding at least one excipient selected from the group consisting of glycine, polysorbate 80, polysorbate 20, and mixtures thereof, prior to lyophilizing the composition. 
   
   
       33 . The method of  claim 32 , wherein said excipient has a pH of about 2.5 to about 3.5. 
   
   
       34 . The method of  claim 32 , wherein said BMP is rhGDF-5. 
   
   
       35 . A device for implanting in a mammal, said device comprising a biocompatible matrix, at least one BMP, and an amount of trehalose sufficient to stabilize said BMP. 
   
   
       36 . The device of  claim 35 , wherein said BMP is rhGDF-5. 
   
   
       37 . The device of  claim 35 , further comprising at least one excipient selected from the group consisting of glycine, polysorbate 80, polysorbate 20, and mixtures thereof. 
   
   
       38 . The device of  claim 37 , wherein said excipient has a pH of about 2.5 to about 3.5. 
   
   
       39 . The device of  claim 35 , wherein said biocompatible matrix is selected from the group consisting of collagen, mineralized collagen, salts of calcium phosphate, ceramics containing calcium, autogenic bone, allogenic bone, xenogenic bone, polylactide (PLA), polyglycolide (PGA), PLA-PGA co-polymers, polycarbonate, polycaprolactone, and mixtures thereof. 
   
   
       40 . The device of  claim 39 , wherein said BMP is rhGDF-5. 
   
   
       41 . The device of  claim 39 , further comprising at least one excipient selected from the group consisting of glycine, polysorbate 80, polysorbate 20, and mixtures thereof. 
   
   
       42 . The device of  claim 41 , wherein said excipient has a pH of about 2.5 to about 3.5. 
   
   
       43 . A surgical kit comprising a biocompatible matrix and a lyophilized BMP composition, wherein said lyophilized BMP composition comprises at least one BMP and an amount of trehalose sufficient to stabilize said BMP. 
   
   
       44 . The surgical kit of  claim 43 , wherein said at least one BMP is rhGDF-5. 
   
   
       45 . The surgical kit of  claim 43 , wherein said biocompatible matrix is sterile. 
   
   
       46 . The surgical kit of  claim 43 , further comprising a container having sterile water. 
   
   
       47 . The surgical kit of  claim 43 , wherein said lyophilized BMP composition further comprises at least one excipient selected from the group consisting of glycine, polysorbate 80, polysorbate 20, and mixtures thereof. 
   
   
       48 . The surgical kit of  claim 47 , wherein said at least one BMP is rhGDF-5. 
   
   
       49 . The surgical kit of  claim 47 , wherein said excipient has a pH of about 2.5 to about 3.5. 
   
   
       50 . A method of treating a patient having a defect comprising the steps of: a) providing a biocompatible matrix having at least one BMP and an amount of trehalose sufficient to stabilize said BMP, and b) implanting said biocompatible matrix into said defect. 
   
   
       51 . The method of  claim 50 , wherein said BMP is rhGDF-5. 
   
   
       52 . The method of  claim 50 , wherein said BMP further comprises at least one excipient selected from the group consisting of glycine, polysorbate 80, polysorbate 20, and mixtures thereof. 
   
   
       53 . The method of  claim 51 , wherein said excipient has a pH of about 2.5 to about 3.5. 
   
   
       54 . The method of  claim 50 , wherein said biocompatible matrix is selected from the group consisting of collagen, mineralized collagen, salts of calcium phosphate, ceramics containing calcium, autogenic bone, allogenic bone, xenogenic bone, polylactide (PLA), polyglycolide (PGA), PLA-PGA co-polymers, polycarbonate, polycaprolactone, and mixtures thereof. 
   
   
       55 . The method of  claim 54 , wherein said BMP is rhGDF-5. 
   
   
       56 . The method of  claim 50 , further comprising the step of adding bone marrow aspirate to said biocompatible matrix. 
   
   
       57 . The method of  claim 50 , wherein said defect is selected from the group consisting of a bone defect, a cartilage defect, a tendon defect, a ligament defect, and an intervertebral disc defect. 
   
   
       58 . A method of treating a patient having a defect comprising the steps of: a) providing a biocompatible matrix, b) providing a BMP having an amount of trehalose sufficient to stabilize said BMP, c) lyophilizing said BMP, d) providing an amount of sterile water sufficient to reconstitute said lyophilized BMP, e) adding said water to said lyophilized BMP to reconstitute said BMP into solution, f) applying said reconstituted BMP solution to said biocompatible matrix, and g) implanting said biocompatible matrix into said patient. 
   
   
       59 . The method of  claim 57 , wherein said BMP is rhGDF-5. 
   
   
       60 . The method of  claim 57 , further comprising the step of adding bone marrow aspirate to said biocompatible matrix. 
   
   
       61 . The method of  claim 57 , further comprising the step of adding at least one excipient selected from the group consisting of glycine, polysorbate 80, polysorbate 20, and mixtures thereof, to said BMP prior to lyophilizing said BMP. 
   
   
       62 . The method of  claim 60 , wherein said excipient has a pH of about 2.5 to about 3.5. 
   
   
       63 . The method of  claim 60 , wherein said BMP is rhGDF-5. 
   
   
       64 . The method of  claim 57 , wherein said biocompatible matrix is selected from the group consisting of collagen, mineralized collagen, salts of calcium phosphate, ceramics containing calcium, autogenic bone, allogenic bone, xenogenic bone, polylactide (PLA), polyglycolide (PGA), PLA-PGA co-polymers, polycarbonate, polycaprolactone, and mixtures thereof. 
   
   
       65 . The method of  claim 57 , wherein said defect is selected from the group consisting of a bone defect, a cartilage defect, a tendon defect, a ligament defect, and an intervertebral disc defect.