CA2386408A1 - Formulations of hyaluronic acid for delivery of osteogenic proteins - Google Patents
Formulations of hyaluronic acid for delivery of osteogenic proteins Download PDFInfo
- Publication number
- CA2386408A1 CA2386408A1 CA002386408A CA2386408A CA2386408A1 CA 2386408 A1 CA2386408 A1 CA 2386408A1 CA 002386408 A CA002386408 A CA 002386408A CA 2386408 A CA2386408 A CA 2386408A CA 2386408 A1 CA2386408 A1 CA 2386408A1
- Authority
- CA
- Canada
- Prior art keywords
- hyaff
- rhbmp
- gel
- hyaluronic acid
- injectable
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Abandoned
Links
- 108090000623 proteins and genes Proteins 0.000 title claims abstract description 46
- 102000004169 proteins and genes Human genes 0.000 title claims abstract description 45
- 239000000203 mixture Substances 0.000 title claims abstract description 39
- 230000002188 osteogenic effect Effects 0.000 title claims abstract description 36
- KIUKXJAPPMFGSW-DNGZLQJQSA-N (2S,3S,4S,5R,6R)-6-[(2S,3R,4R,5S,6R)-3-Acetamido-2-[(2S,3S,4R,5R,6R)-6-[(2R,3R,4R,5S,6R)-3-acetamido-2,5-dihydroxy-6-(hydroxymethyl)oxan-4-yl]oxy-2-carboxy-4,5-dihydroxyoxan-3-yl]oxy-5-hydroxy-6-(hydroxymethyl)oxan-4-yl]oxy-3,4,5-trihydroxyoxane-2-carboxylic acid Chemical compound CC(=O)N[C@H]1[C@H](O)O[C@H](CO)[C@@H](O)[C@@H]1O[C@H]1[C@H](O)[C@@H](O)[C@H](O[C@H]2[C@@H]([C@@H](O[C@H]3[C@@H]([C@@H](O)[C@H](O)[C@H](O3)C(O)=O)O)[C@H](O)[C@@H](CO)O2)NC(C)=O)[C@@H](C(O)=O)O1 KIUKXJAPPMFGSW-DNGZLQJQSA-N 0.000 title claims abstract description 18
- 229920002674 hyaluronan Polymers 0.000 title claims abstract description 17
- 229960003160 hyaluronic acid Drugs 0.000 title claims abstract description 17
- 238000009472 formulation Methods 0.000 title abstract description 21
- 239000001506 calcium phosphate Substances 0.000 claims abstract description 40
- QORWJWZARLRLPR-UHFFFAOYSA-H tricalcium bis(phosphate) Chemical compound [Ca+2].[Ca+2].[Ca+2].[O-]P([O-])([O-])=O.[O-]P([O-])([O-])=O QORWJWZARLRLPR-UHFFFAOYSA-H 0.000 claims abstract description 40
- 229910000391 tricalcium phosphate Inorganic materials 0.000 claims abstract description 40
- 229940078499 tricalcium phosphate Drugs 0.000 claims abstract description 40
- 235000019731 tricalcium phosphate Nutrition 0.000 claims abstract description 40
- 238000000034 method Methods 0.000 claims abstract description 14
- 239000011148 porous material Substances 0.000 claims description 25
- 239000000969 carrier Substances 0.000 claims description 8
- 239000002904 solvent Substances 0.000 claims description 5
- 230000036571 hydration Effects 0.000 claims description 4
- 238000006703 hydration reaction Methods 0.000 claims description 4
- 108010049931 Bone Morphogenetic Protein 2 Proteins 0.000 claims description 3
- 102100024506 Bone morphogenetic protein 2 Human genes 0.000 claims description 3
- 239000011159 matrix material Substances 0.000 claims description 3
- 230000001131 transforming effect Effects 0.000 claims description 3
- 239000000499 gel Substances 0.000 abstract description 66
- 239000007972 injectable composition Substances 0.000 abstract description 7
- 235000018102 proteins Nutrition 0.000 description 34
- 210000000988 bone and bone Anatomy 0.000 description 28
- 229920001223 polyethylene glycol Polymers 0.000 description 16
- 230000014759 maintenance of location Effects 0.000 description 13
- 238000001727 in vivo Methods 0.000 description 11
- 150000002148 esters Chemical class 0.000 description 10
- 230000011164 ossification Effects 0.000 description 10
- 239000007787 solid Substances 0.000 description 10
- 230000007547 defect Effects 0.000 description 9
- 238000000338 in vitro Methods 0.000 description 8
- 206010017076 Fracture Diseases 0.000 description 7
- 239000000872 buffer Substances 0.000 description 7
- SECXISVLQFMRJM-UHFFFAOYSA-N N-Methylpyrrolidone Chemical compound CN1CCCC1=O SECXISVLQFMRJM-UHFFFAOYSA-N 0.000 description 6
- 241000283973 Oryctolagus cuniculus Species 0.000 description 6
- UIIMBOGNXHQVGW-UHFFFAOYSA-M Sodium bicarbonate Chemical compound [Na+].OC([O-])=O UIIMBOGNXHQVGW-UHFFFAOYSA-M 0.000 description 6
- 210000000845 cartilage Anatomy 0.000 description 6
- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid Chemical compound OC(=O)CC(O)(C(O)=O)CC(O)=O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 description 6
- 230000000717 retained effect Effects 0.000 description 6
- 208000010392 Bone Fractures Diseases 0.000 description 5
- 241000700159 Rattus Species 0.000 description 5
- 229920000642 polymer Polymers 0.000 description 5
- BVKZGUZCCUSVTD-UHFFFAOYSA-M Bicarbonate Chemical compound OC([O-])=O BVKZGUZCCUSVTD-UHFFFAOYSA-M 0.000 description 4
- -1 hyaluronic acid benzyl ester Chemical class 0.000 description 4
- 238000002347 injection Methods 0.000 description 4
- 239000007924 injection Substances 0.000 description 4
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 description 4
- 239000000463 material Substances 0.000 description 4
- 238000002156 mixing Methods 0.000 description 4
- 239000003960 organic solvent Substances 0.000 description 4
- 239000002245 particle Substances 0.000 description 4
- 239000000700 radioactive tracer Substances 0.000 description 4
- 208000017013 ulna fracture Diseases 0.000 description 4
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 3
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 3
- 239000002202 Polyethylene glycol Substances 0.000 description 3
- 208000027418 Wounds and injury Diseases 0.000 description 3
- 238000003556 assay Methods 0.000 description 3
- 230000003115 biocidal effect Effects 0.000 description 3
- 239000003795 chemical substances by application Substances 0.000 description 3
- 230000006378 damage Effects 0.000 description 3
- 239000007943 implant Substances 0.000 description 3
- 208000014674 injury Diseases 0.000 description 3
- 230000004048 modification Effects 0.000 description 3
- 238000012986 modification Methods 0.000 description 3
- 238000002360 preparation method Methods 0.000 description 3
- 229910000030 sodium bicarbonate Inorganic materials 0.000 description 3
- 235000017557 sodium bicarbonate Nutrition 0.000 description 3
- 210000000623 ulna Anatomy 0.000 description 3
- FJQZXCPWAGYPSD-UHFFFAOYSA-N 1,3,4,6-tetrachloro-3a,6a-diphenylimidazo[4,5-d]imidazole-2,5-dione Chemical compound ClN1C(=O)N(Cl)C2(C=3C=CC=CC=3)N(Cl)C(=O)N(Cl)C12C1=CC=CC=C1 FJQZXCPWAGYPSD-UHFFFAOYSA-N 0.000 description 2
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 2
- 208000003044 Closed Fractures Diseases 0.000 description 2
- CEAZRRDELHUEMR-URQXQFDESA-N Gentamicin Chemical compound O1[C@H](C(C)NC)CC[C@@H](N)[C@H]1O[C@H]1[C@H](O)[C@@H](O[C@@H]2[C@@H]([C@@H](NC)[C@@](C)(O)CO2)O)[C@H](N)C[C@@H]1N CEAZRRDELHUEMR-URQXQFDESA-N 0.000 description 2
- 229930182566 Gentamicin Natural products 0.000 description 2
- 206010031252 Osteomyelitis Diseases 0.000 description 2
- UIIMBOGNXHQVGW-DEQYMQKBSA-M Sodium bicarbonate-14C Chemical compound [Na+].O[14C]([O-])=O UIIMBOGNXHQVGW-DEQYMQKBSA-M 0.000 description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 2
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 2
- 229930006000 Sucrose Natural products 0.000 description 2
- 239000003242 anti bacterial agent Substances 0.000 description 2
- 229940088710 antibiotic agent Drugs 0.000 description 2
- 239000012062 aqueous buffer Substances 0.000 description 2
- 230000002146 bilateral effect Effects 0.000 description 2
- 230000015572 biosynthetic process Effects 0.000 description 2
- OSGAYBCDTDRGGQ-UHFFFAOYSA-L calcium sulfate Inorganic materials [Ca+2].[O-]S([O-])(=O)=O OSGAYBCDTDRGGQ-UHFFFAOYSA-L 0.000 description 2
- ZOMBKNNSYQHRCA-UHFFFAOYSA-J calcium sulfate hemihydrate Chemical compound O.[Ca+2].[Ca+2].[O-]S([O-])(=O)=O.[O-]S([O-])(=O)=O ZOMBKNNSYQHRCA-UHFFFAOYSA-J 0.000 description 2
- 230000022159 cartilage development Effects 0.000 description 2
- 210000004027 cell Anatomy 0.000 description 2
- 229940079593 drug Drugs 0.000 description 2
- 239000003814 drug Substances 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 239000012894 fetal calf serum Substances 0.000 description 2
- 239000011507 gypsum plaster Substances 0.000 description 2
- JYGXADMDTFJGBT-VWUMJDOOSA-N hydrocortisone Chemical compound O=C1CC[C@]2(C)[C@H]3[C@@H](O)C[C@](C)([C@@](CC4)(O)C(=O)CO)[C@@H]4[C@@H]3CCC2=C1 JYGXADMDTFJGBT-VWUMJDOOSA-N 0.000 description 2
- 229910052588 hydroxylapatite Inorganic materials 0.000 description 2
- 238000011065 in-situ storage Methods 0.000 description 2
- 230000006698 induction Effects 0.000 description 2
- 230000001939 inductive effect Effects 0.000 description 2
- 208000015181 infectious disease Diseases 0.000 description 2
- 239000004310 lactic acid Substances 0.000 description 2
- 235000014655 lactic acid Nutrition 0.000 description 2
- 238000011587 new zealand white rabbit Methods 0.000 description 2
- 230000002138 osteoinductive effect Effects 0.000 description 2
- XYJRXVWERLGGKC-UHFFFAOYSA-D pentacalcium;hydroxide;triphosphate Chemical compound [OH-].[Ca+2].[Ca+2].[Ca+2].[Ca+2].[Ca+2].[O-]P([O-])([O-])=O.[O-]P([O-])([O-])=O.[O-]P([O-])([O-])=O XYJRXVWERLGGKC-UHFFFAOYSA-D 0.000 description 2
- 239000008194 pharmaceutical composition Substances 0.000 description 2
- 238000001556 precipitation Methods 0.000 description 2
- 108090000765 processed proteins & peptides Proteins 0.000 description 2
- 102000004196 processed proteins & peptides Human genes 0.000 description 2
- 238000010188 recombinant method Methods 0.000 description 2
- 210000002966 serum Anatomy 0.000 description 2
- 230000007928 solubilization Effects 0.000 description 2
- 238000005063 solubilization Methods 0.000 description 2
- 239000000243 solution Substances 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 239000005720 sucrose Substances 0.000 description 2
- 229960003165 vancomycin Drugs 0.000 description 2
- MYPYJXKWCTUITO-LYRMYLQWSA-N vancomycin Chemical compound O([C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1OC1=C2C=C3C=C1OC1=CC=C(C=C1Cl)[C@@H](O)[C@H](C(N[C@@H](CC(N)=O)C(=O)N[C@H]3C(=O)N[C@H]1C(=O)N[C@H](C(N[C@@H](C3=CC(O)=CC(O)=C3C=3C(O)=CC=C1C=3)C(O)=O)=O)[C@H](O)C1=CC=C(C(=C1)Cl)O2)=O)NC(=O)[C@@H](CC(C)C)NC)[C@H]1C[C@](C)(N)[C@H](O)[C@H](C)O1 MYPYJXKWCTUITO-LYRMYLQWSA-N 0.000 description 2
- 102100025908 5-oxoprolinase Human genes 0.000 description 1
- FHVDTGUDJYJELY-UHFFFAOYSA-N 6-{[2-carboxy-4,5-dihydroxy-6-(phosphanyloxy)oxan-3-yl]oxy}-4,5-dihydroxy-3-phosphanyloxane-2-carboxylic acid Chemical compound O1C(C(O)=O)C(P)C(O)C(O)C1OC1C(C(O)=O)OC(OP)C(O)C1O FHVDTGUDJYJELY-UHFFFAOYSA-N 0.000 description 1
- 108020004491 Antisense DNA Proteins 0.000 description 1
- 108020000948 Antisense Oligonucleotides Proteins 0.000 description 1
- 108010007726 Bone Morphogenetic Proteins Proteins 0.000 description 1
- 102000007350 Bone Morphogenetic Proteins Human genes 0.000 description 1
- 208000017234 Bone cyst Diseases 0.000 description 1
- 208000020084 Bone disease Diseases 0.000 description 1
- 108090000654 Bone morphogenetic protein 1 Proteins 0.000 description 1
- 102100028728 Bone morphogenetic protein 1 Human genes 0.000 description 1
- 229920002134 Carboxymethyl cellulose Polymers 0.000 description 1
- 102000008186 Collagen Human genes 0.000 description 1
- 108010035532 Collagen Proteins 0.000 description 1
- 102000004127 Cytokines Human genes 0.000 description 1
- 108090000695 Cytokines Proteins 0.000 description 1
- 108020004414 DNA Proteins 0.000 description 1
- 241000722985 Fidia Species 0.000 description 1
- WHUUTDBJXJRKMK-UHFFFAOYSA-N Glutamic acid Natural products OC(=O)C(N)CCC(O)=O WHUUTDBJXJRKMK-UHFFFAOYSA-N 0.000 description 1
- 101000720962 Homo sapiens 5-oxoprolinase Proteins 0.000 description 1
- 206010020649 Hyperkeratosis Diseases 0.000 description 1
- 241000906034 Orthops Species 0.000 description 1
- 208000001132 Osteoporosis Diseases 0.000 description 1
- 102000007056 Recombinant Fusion Proteins Human genes 0.000 description 1
- 108010008281 Recombinant Fusion Proteins Proteins 0.000 description 1
- QAOWNCQODCNURD-UHFFFAOYSA-L Sulfate Chemical compound [O-]S([O-])(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-L 0.000 description 1
- 241000897276 Termes Species 0.000 description 1
- 102000009618 Transforming Growth Factors Human genes 0.000 description 1
- 108010009583 Transforming Growth Factors Proteins 0.000 description 1
- 108010059993 Vancomycin Proteins 0.000 description 1
- 229920006222 acrylic ester polymer Polymers 0.000 description 1
- 239000000654 additive Substances 0.000 description 1
- 230000002411 adverse Effects 0.000 description 1
- 229940072056 alginate Drugs 0.000 description 1
- 229920000615 alginic acid Polymers 0.000 description 1
- 235000010443 alginic acid Nutrition 0.000 description 1
- 210000001909 alveolar process Anatomy 0.000 description 1
- 239000003816 antisense DNA Substances 0.000 description 1
- 239000000074 antisense oligonucleotide Substances 0.000 description 1
- 238000012230 antisense oligonucleotides Methods 0.000 description 1
- 230000003416 augmentation Effects 0.000 description 1
- 125000001797 benzyl group Chemical group [H]C1=C([H])C([H])=C(C([H])=C1[H])C([H])([H])* 0.000 description 1
- 229920001222 biopolymer Polymers 0.000 description 1
- 229940112869 bone morphogenetic protein Drugs 0.000 description 1
- 230000010478 bone regeneration Effects 0.000 description 1
- 239000001768 carboxy methyl cellulose Substances 0.000 description 1
- 235000010948 carboxy methyl cellulose Nutrition 0.000 description 1
- 239000008112 carboxymethyl-cellulose Substances 0.000 description 1
- 230000015556 catabolic process Effects 0.000 description 1
- 210000000038 chest Anatomy 0.000 description 1
- 239000011248 coating agent Substances 0.000 description 1
- 238000000576 coating method Methods 0.000 description 1
- 229920001436 collagen Polymers 0.000 description 1
- 239000000515 collagen sponge Substances 0.000 description 1
- 239000002299 complementary DNA Substances 0.000 description 1
- 239000002131 composite material Substances 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 238000006731 degradation reaction Methods 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 231100000673 dose–response relationship Toxicity 0.000 description 1
- 230000032050 esterification Effects 0.000 description 1
- 238000005886 esterification reaction Methods 0.000 description 1
- 230000001747 exhibiting effect Effects 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 239000000835 fiber Substances 0.000 description 1
- 239000012634 fragment Substances 0.000 description 1
- 230000004927 fusion Effects 0.000 description 1
- 235000013922 glutamic acid Nutrition 0.000 description 1
- 239000004220 glutamic acid Substances 0.000 description 1
- 150000004676 glycans Polymers 0.000 description 1
- 230000012010 growth Effects 0.000 description 1
- 239000003102 growth factor Substances 0.000 description 1
- 230000035876 healing Effects 0.000 description 1
- 229960000890 hydrocortisone Drugs 0.000 description 1
- 230000036512 infertility Effects 0.000 description 1
- 229910052500 inorganic mineral Inorganic materials 0.000 description 1
- 230000010354 integration Effects 0.000 description 1
- 210000003041 ligament Anatomy 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 230000010874 maintenance of protein location Effects 0.000 description 1
- 238000013289 male long evans rat Methods 0.000 description 1
- 125000002496 methyl group Chemical group [H]C([H])([H])* 0.000 description 1
- 239000003094 microcapsule Substances 0.000 description 1
- 239000011859 microparticle Substances 0.000 description 1
- 239000004005 microsphere Substances 0.000 description 1
- 239000011707 mineral Substances 0.000 description 1
- 235000010755 mineral Nutrition 0.000 description 1
- 230000001089 mineralizing effect Effects 0.000 description 1
- 230000000921 morphogenic effect Effects 0.000 description 1
- 210000000963 osteoblast Anatomy 0.000 description 1
- 210000005009 osteogenic cell Anatomy 0.000 description 1
- 230000001009 osteoporotic effect Effects 0.000 description 1
- 230000003239 periodontal effect Effects 0.000 description 1
- 229920001184 polypeptide Polymers 0.000 description 1
- 150000004804 polysaccharides Polymers 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 230000002265 prevention Effects 0.000 description 1
- 230000002035 prolonged effect Effects 0.000 description 1
- 239000002510 pyrogen Substances 0.000 description 1
- 238000011552 rat model Methods 0.000 description 1
- 238000004626 scanning electron microscopy Methods 0.000 description 1
- 239000003352 sequestering agent Substances 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 210000000130 stem cell Anatomy 0.000 description 1
- 238000013268 sustained release Methods 0.000 description 1
- 239000012730 sustained-release form Substances 0.000 description 1
- 210000002435 tendon Anatomy 0.000 description 1
- 210000001519 tissue Anatomy 0.000 description 1
- 230000000451 tissue damage Effects 0.000 description 1
- 231100000827 tissue damage Toxicity 0.000 description 1
- MYPYJXKWCTUITO-UHFFFAOYSA-N vancomycin Natural products O1C(C(=C2)Cl)=CC=C2C(O)C(C(NC(C2=CC(O)=CC(O)=C2C=2C(O)=CC=C3C=2)C(O)=O)=O)NC(=O)C3NC(=O)C2NC(=O)C(CC(N)=O)NC(=O)C(NC(=O)C(CC(C)C)NC)C(O)C(C=C3Cl)=CC=C3OC3=CC2=CC1=C3OC1OC(CO)C(O)C(O)C1OC1CC(C)(N)C(O)C(C)O1 MYPYJXKWCTUITO-UHFFFAOYSA-N 0.000 description 1
- 239000003981 vehicle Substances 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/0012—Galenical forms characterised by the site of application
- A61K9/0019—Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner
- A61K9/0024—Solid, semi-solid or solidifying implants, which are implanted or injected in body tissue
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
- A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- A61K38/18—Growth factors; Growth regulators
- A61K38/1875—Bone morphogenic factor; Osteogenins; Osteogenic factor; Bone-inducing factor
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
- A61K47/26—Carbohydrates, e.g. sugar alcohols, amino sugars, nucleic acids, mono-, di- or oligo-saccharides; Derivatives thereof, e.g. polysorbates, sorbitan fatty acid esters or glycyrrhizin
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/30—Macromolecular organic or inorganic compounds, e.g. inorganic polyphosphates
- A61K47/36—Polysaccharides; Derivatives thereof, e.g. gums, starch, alginate, dextrin, hyaluronic acid, chitosan, inulin, agar or pectin
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/0012—Galenical forms characterised by the site of application
- A61K9/0019—Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/14—Macromolecular materials
- A61L27/20—Polysaccharides
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/14—Macromolecular materials
- A61L27/22—Polypeptides or derivatives thereof, e.g. degradation products
- A61L27/227—Other specific proteins or polypeptides not covered by A61L27/222, A61L27/225 or A61L27/24
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/40—Composite materials, i.e. containing one material dispersed in a matrix of the same or different material
- A61L27/44—Composite materials, i.e. containing one material dispersed in a matrix of the same or different material having a macromolecular matrix
- A61L27/46—Composite materials, i.e. containing one material dispersed in a matrix of the same or different material having a macromolecular matrix with phosphorus-containing inorganic fillers
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P19/00—Drugs for skeletal disorders
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P19/00—Drugs for skeletal disorders
- A61P19/08—Drugs for skeletal disorders for bone diseases, e.g. rachitism, Paget's disease
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L2300/00—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices
- A61L2300/40—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices characterised by a specific therapeutic activity or mode of action
- A61L2300/412—Tissue-regenerating or healing or proliferative agents
- A61L2300/414—Growth factors
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L2400/00—Materials characterised by their function or physical properties
- A61L2400/06—Flowable or injectable implant compositions
Abstract
An injectable formulation is disclosed for delivery of osteogenic proteins. The formulation comprises a pharmaceutically acceptable admixture of an osteogenic protein; and formulations comprising osteogenic protein, hyaluronic acid derivatives and tricalcium phosphate are also disclosed. Methods for formulating porous injectable gels and pastes from hyaluronic acid are also disclosed.
Description
FORMULATIONS OF HYALURONIC ACID FOR DELIVERY OF OSTEOGENIC PROTEINS
BACKGROUND OF THE INVENTION
The subject invention relates to the field of osteogenic proteins and pharmaceutical formulations thereof. More particularly, the subject invention involves injectable pharmaceutical formulations comprising hyaluronic acid derivitives and osteogenic proteins.
The invention further provides methods for formulating porous injectable gels and pastes from hyaluronic acid.
Osteogenic proteins are those proteins capable of inducing, or assisting in the induction of, cartilage and/or bone formation. Many such osteogenic proteins have in recent years been isolated and characterized, and some have been produced by recombinant methods. For example, so-called bone morphogenic proteins (BMP) have been isolated from demineralized bone tissue (see e.g. Urist US 4,455,256); a number of such BMP
proteins have been produced by recombinant techniques (see e.g. Wang et al. US
4,877,864 and Wang et al. US 5,013,549); a family of transforming growth factors (TGF-oc and TGF-(3) has been identified as potentially useful in the treatment of bone disease (see e.g.
Derynck et al., EP
154,434); a protein designated Vgr-1 has been found to be expressed at high levels in osteogenic cells (see Lyons et al. (1989) Proc. Nat'1. Acad. Sci. USA 86, 4554-4558); and proteins designated OP-1, COP-5 and COP-7 have purportedly shown bone inductive activity (see Oppermann, et al. U.S. 5,001,691).
Various formulations designed to deliver osteogenic proteins to a site where induction of bone formation is desired have been developed. For example, certain polymeric matrices such as acrylic ester polymer (Urist, US 4,526,909) and lactic acid polymer (Urist, US
BACKGROUND OF THE INVENTION
The subject invention relates to the field of osteogenic proteins and pharmaceutical formulations thereof. More particularly, the subject invention involves injectable pharmaceutical formulations comprising hyaluronic acid derivitives and osteogenic proteins.
The invention further provides methods for formulating porous injectable gels and pastes from hyaluronic acid.
Osteogenic proteins are those proteins capable of inducing, or assisting in the induction of, cartilage and/or bone formation. Many such osteogenic proteins have in recent years been isolated and characterized, and some have been produced by recombinant methods. For example, so-called bone morphogenic proteins (BMP) have been isolated from demineralized bone tissue (see e.g. Urist US 4,455,256); a number of such BMP
proteins have been produced by recombinant techniques (see e.g. Wang et al. US
4,877,864 and Wang et al. US 5,013,549); a family of transforming growth factors (TGF-oc and TGF-(3) has been identified as potentially useful in the treatment of bone disease (see e.g.
Derynck et al., EP
154,434); a protein designated Vgr-1 has been found to be expressed at high levels in osteogenic cells (see Lyons et al. (1989) Proc. Nat'1. Acad. Sci. USA 86, 4554-4558); and proteins designated OP-1, COP-5 and COP-7 have purportedly shown bone inductive activity (see Oppermann, et al. U.S. 5,001,691).
Various formulations designed to deliver osteogenic proteins to a site where induction of bone formation is desired have been developed. For example, certain polymeric matrices such as acrylic ester polymer (Urist, US 4,526,909) and lactic acid polymer (Urist, US
4,563,489) have been utilized.
A biodegradable matrix of porous particles for delivery of an osteogenic protein designated as OP is disclosed in Kuber A. Sampath, U.S. 5,108,753.
Brekke et al., United States Patents 4,186,448 and 5,133,755 describe methods of forming highly porous biodegradable materials composed of polymers of lactic acid ("OPLA") Okada et al., US 4,652,441, US 4,711,782, US 4,917,893 and US 5,061,492 and Yamamoto et al., US 4,954,298 disclose a prolonged-release microcapsule comprising a polypeptide drug and a drug-retaining substance encapsulated in an inner aqueous layer surrounded by a polymer wall substance in an outer oil layer.
Yamazaki et al., Clin. Orthop. and Related Research, 234:240-249 (1988) disclose the use of implants comprising 1 mg of bone morphogenetic protein purified from bone and S mg of Plaster of Paris. United States Patent 4,645,503 discloses composites of hydroxyapatite and Plaster of Paris as bone implant materials.
Collagen matrices have also been used as delivery vehicles for osteogenic proteins (see e.g. Jeffries, U.S. 4,394,370).
SUMMARY OF THE INVENTION
The present invention provides injectable formulations for delivery of osteogenic proteins. In one embodiment the composition comprises the osteogenic protein and hyaluronic acid esters. In another embodiment, the composition may further include tricalcium phosphate. The injectable formulations of the invention allows for closed fracture repair and other skeletal tissue without an open reduction procedure as is necessary with implantable devices.
The present invention further provides methods for preparing injectable gels or pastes useful as a carrier for osteogenic proteins by transforming various non-woven pads and sponges of hyaluronic acid benzyl ester into injectable gel or paste formulations by hydration or solvent addition. In another embodiment, the invention comprises compositions comprising the transformed injectable gel or paste formulations.
The methods and compositions of the present invention are useful for the preparation of formulations of osteoinductive proteins which can be used, among other uses, to promote the formation of cartilage and/or bone, for repair of tissue damage and fractures. The invention further provides methods for treating patients in need of cartilage and/or bone repair and/or growth.
_J_ BRIEF DESCRIPTION OF THE DRAWINGS
Figure 1 sets forth in vitro release kinetics of'z5I-rhBMP-2 in Hyaff gels.
Figure 2 sets forth in vivo retention of lzsl_rhBMP-2 in Hyaff 11/PEG, ACS, and buffer.
Figure 3 sets forth in vitro release kinetics of'z5I-rhBMP-2 in Hyaff gels/TCP.
S Figure 4 sets forth in vivo biodistribution of'zsI_rhBMP-2.
DETAILED DESCRIPTION OF THE INVENTION
The invention provides injectable formulations for delivery of osteogenic proteins.
The compositions comprise an injectable formulation of hyaluronic acid esters and osteogenic protein. The present invention further provides processes for preparing injectable gel or paste formulations by transforming various non-woven pads and sponges of hyaluronic acid benzyl ester by hydration or solvent addition yielding gels with in vivo residence times from days to up to several months. Total or partial esters of hyaluronic acid are described in US 5,336,767.
Partial esters of Hyaff solids are transformed into gels using aqueous buffer or organic solvents (such as N-methyl pyrrolidinone, dimethyl sulfoxide, etc), while complete esters of Hyaff solids are transformed into gels using organic solvents. In other embodiments pore formers may be introduced to the solublized carriers to increase porosity. The addition of pore formers would allow in situ pore formation after injection in vivo by solubilization of pore former and precipitation/phase inversion of Garner. Suitable liquid pore formers include polyethylene glycol or PEG at 10 - 90 % volume per volume ratios) and solid pore formers (such as sodium bicarbonate, sodium chloride, citric acid, sucrose, etc., at 1:1 - 21:1 pore former:Hyaff weight per weight ratios) to increase porosity. The gel/paste can also contain TCP (tri-calcium phosphate) particles as a mineral component for example, at 0.1 - 100 weight per volume range.
The amount , type and size of the pore forming agent is optimized to leave voids sufficient for cell ingrowth into injectable gel when pore forming agent and solvent are extracted from the carrier in vivo by solubilization of pore forming agent and precipitation/phase inversion of carrier in situ.
The osteogenic proteins useful with the injectable carriers made in accordance with the subject invention are well known to those skilled in the art and include those discussed above. The preferred osteogenic proteins for use herein are those of the BMP
class identified as BMP-1 through BMP-12 in US 4,877,864; US 5,013,649; WO 90/11366 published October 4, 1990; WO 91/18098 published November 28, 1991; WO 93/00432, published January 7, 1993; United States Serial Numbers 08/247,908 and 08/247,904, both filed May 20, 1994; and United States Serial Number 08/217,780, filed on March 25, 1994.
The disclosure of the above publications are hereby incorporated by reference. The most preferred is BMP-2, the full length cDNA sequence of which is described in detail in the '649 patent.
Of course, combinations of two or more of such osteogenic proteins may be used, as may fragments of such proteins that also exhibit osteogenic activity. Such osteogenic proteins are known to be homodimeric species, but also exhibit activity as mixed heterodimers.
Heterodimeric forms of osteogenic proteins may also be used in the practice of the subject invention. BMP heterodimers are described in W093/09229, the disclosure of which is hereby incorporated by reference. Recombinant proteins are preferred over naturally occurnng isolated proteins. The amount of osteogenic protein useful herein is that amount effective to stimulate increased osteogenic activity of infiltrating progenitor cells, and will depend upon the size and nature of defect being treated as well as the carrier being employed.
The formulations may be injected for example into tendons, damaged cartilage tissue, ligaments, and/or their attachment sites to bones. Injectable formulations may also fmd application to other bone sites such as bone cysts, bone defects, intraosseous sites and closed fractures.
The dosage regimen will be determined by the clinical indication being addressed, as well as by various patient variables (e.g. weight, age, sex) and clinical presentation (e.g.
extent of injury, site of injury, etc.). In general, the dosage of osteogenic protein will'be in the range of from about 0.1 to 4 mg/ml.
The injectable osteogenic protein formulations may be provided to the clinic as a single formulation, or the formulation may be provided as a multicomponent kit wherein, e.g.
the osteogenic protein is provided in one vial and the injectable hyaluronic paste is provided separately.
The compositions of the subject invention allow therapeutically effective amounts of osteoinductive protein to be delivered to an injury site where cartilage and/or bone formation is desired. The formulations may be used as a substitute for autologous bone graft in fresh and non-union fractures, spinal fusions, and bone defect repair in the orthopaedic field; in _j_ cranio/maxillofacial reconstructions; for prosthesis integration, especially as a surface coating to improve fixation of prosthetic implants such as hydroxyapatite coated prostheses; in osteomyelitis for bone regeneration; and in the dental field for augmentation of the alveolar ridge and periodontal defects and tooth extraction sockets. The methods and formulations of the present invention may be useful in the treatment and/or prevention of osteoporosis, or the treatment of osteoporotic or osteopenic bone. In another embodiment, formulations of the present invention may be used in the process known as distraction osteogenesis. When used to treat osteomyelitis or for bone repair with minimal infection, the osteogenic protein may be used in combination with porous microparticles and antibiotics, with the addition of protein sequestering agents such as alginate, cellulosics, especially carboxymethylcellulose, diluted using aqueous glycerol. The antibiotic is selected for its ability to decrease infection while having minimal adverse effects on bone formation. Preferred antibiotics for use in the devices of the present invention include vancomycin and gentamycin. The antibiotic may be in any pharmaceutically acceptable form, such as vancomycin HCl or gentamycin sulfate. The antibiotic is preferably present in a concentration of from about 0.1 mg/mL to about 10.0 mg/mL. The traditional preparation of formulations in pharmaceutically acceptable form (i.e.
pyrogen free, appropriate pH and isotonicity, sterility, etc.) is well within the skill in the art and is applicable to the formulations of the invention.
Hyaluronic derivitive compositions of the invention prepared by hydration or solvent addition of insoluble or partially soluble non- woven pads or sponges may also be ultilized in combination with other drugs, growth factors, peptides, proteins, cytokines, oligonucleeotides antisense oligonucleotides, DNA and polymers. These compounds may be added by mixing them with the carriers. Or by covalent attachment to the polymer carriers. The hyaluronic derivitive compositions may also be used with DNA encoding for BMPs and cells transduced or transfected with genes encoding BMP proteins.
The following examples are illustrative of the present invention and are not limiting in any manner. Modifications, variations and minor enhancements are contemplated and are within the present invention.
PREPARATION OF INJECTABLE HYALURONIC ACID ESTERS
The starting Hyaff hyaluronic acid (Fidia Advanced Biopolymers, Abano Terme, Italy) materials are solids such as non-woven pads, felts, sheets, powders, sponges, and microspheres. The Hyaff materials are esters of hyaluronic acid exhibiting various ester moities (e.g., benzyl, ethyl,propyl pentyl or larger molecules such as hydrocortisone or methyl prednislone, etc.) as well as various degrees of esterification (i.e., partial esters or complete esters). Partial esters of Hyaff are designated by percent esterfication ranging from 50-99 (e.g., Hyaff 11p65, Hyaff 11p80, etc.), while complete esters are 100 % esters of hyaluronic acid (e.g., Hyaff 11).
Hyaff gel classification used in supporting data is as follows and is followed by examples of select formulations:
- Hyaff 11 gel: Hyaff 11 non-woven pad transformed into gel with organic solvent to yield 10 % solids - Hyaff 11/bicarbonate gel: Hyaff 11 gel mixed with sodium bicarbonate as pore former at 15:1 (w/w) bicarbonate to Hyaff 11 - Hyaff 11/PEG gel: Hyaff 11 gel mixed with polyethylene glycol(200mw) as pore former at 33 - 50 % (v/v) range - Hyaff 11/TCP gel: Hyaff 11 gel mixed with 30 % w/v TCP
- Hyaff 11/bicarbonate/TCP gel: Hyaff 11/bicarbonate gel mixed with 30 % w/v TCP
- Hyaff 11/PEG/TCP gel: Hyaff 11/PEG gel mixed with 30 % w/v TCP
- Hyaff 11p80 gel: Hyaff 11p80 non-woven pad transformed into gel with organic solvent to yield 5 % solids - Hyaff 11p65 gel: Hyaff 11p65 non-woven pad hydrated with aqueous buffer to yield 6 -15 % solids - Hyaff 11p65/TCP gel: Hyaff 11p65 gel mixed with 30 % w/v TCP
Hyaff 11p65 non-woven pads were hydrated with glutamic acid buffer (pH 4.5) containing rhBMP-2 (0.1 mg/mL final cone) to yield either 6 % - 15 % solids (w/v) and mixed thoroughly to form a paste. Hyaff 11p80 and Hyaff 11 non-woven pads were solubilized in N-methyl-pyrrolidinone (NMP) or dimethyl sulfoxide(DMSO)to yield a 1 - 30 _7_ w/v solution. These solutions were then mixed with either rhBMP-2-containing buffer (10 v/v, 0.1 mg/mL rhBMP-2), or lyophilized rhBMP-2 (0.1 mg/mL) followed by the addition of various pore formers (polyethylene glycol, sodium bicarbonate, sucrose, NaCI, citric acid)and tricalcium phosphate (TCP). Particle size of solid pore formers and TCP used was <
600um, preferably <200umLiquid pore formers such as PEG(200mw) were mixed at 10-90%
v/v ratios, and solid pore formers were mixed at 9:1 - 21:1 (w/w) pore former to carrier ratios.
TCP was mixed at 0.1-30% (w/v). TCP (45-125 micron particle size) was mixed thoroughly into rhBMP-2/Hyaff 11 or rhBMP-2/Hyaff 11p65 gel at 30% (w/v). Separately, rhBMP-2 was adsorbed onto TCP first, followed by mixing with Hyaff 11 or Hyaff 11p65 gel.Formulations were chosen based on injectability through an 18 g needle.
Microstructure was characterized by scanning electron microscopy (SEM).
SEMS revealed varying degrees of pore structure and porosity. Hyaff 11p65 6%
gel exhibited longer fibers than the 15% formulation; with both displaying a high level of porosity. Both Hyaff 11 and Hyaff 11p80 gels showed minimal pore structure and porosity, whereas those carriers with pore formers displayed a high level of porosity.
Pore formers and/
or additives that yielded injectable mixtures were PEG, sodium bicarbonate and TCP.
IN VITRO RELEASE KINETICS
rhBMP-2 was radiolabeled with'zsI using the Iodogen method (Pierce) and used as a tracer for 0.1 mg/ml rhBMP-2 delivered in 100 u1 Hyaff=11p65 gel, Hyaff 11p80 gel, Hyaff 11 gel and Hyaff 11lPEG (n=4). 'z5I-rhBMP-2 loaded samples (50,000 cpm/sample) were incubated in 1 ml fetal calf serum (Hyclone) at 37°C on a shaker, and radioactivity of the Garner measured up to 14 days using a gamma counter. Fresh serum was replaced after each time point. 'z5I-rhBMP-2 release from injectable formulations were compared to those of implantable sponges and pads of Hyaff 11 and Hyaff 11p80.
Auto cross-linked polysaccharide form of derivitized hyaluronic acid, ACP gel, is used for the in vitro release study and the rat ectopic assay. For the in vitro release study, 2 ml ACP gel is mixed with 1.53 mg rhBMP-2 cake (which corresponds to 0.2 mg actual rhBMP-2 at 8 mg rhBMP-2 per 61 mg cake weight) and'z5I-rhBMP-2 (100 ml total, 20 mCi/200 ml gel) and drawn up into 1 ml syringes resulting in approximately 10 % gel dilution. ACP
gel for the rat _g_ ectopic study does not contain the tracer but is diluted with MRF-00906 buffer. 200 ml injections are performed using a 22 gauge needle. The final concentration of rhBMP-2 will be 0.1 mg/ml, or 20 mg per 200 ml injection. The final concentration of '251-rhBMP-2 will be approximately 20 mCi per 200 ml injection. The ACP gel will be injected at room temperature.
In vitro release kinetics showed greatest retention of rhBMP-2 over the 2 weeks in the Hyaff 11/PEG gel followed by Hyaff 11p80 gel and Hyaff 11 gel (Fig. 1). Hyaff 11p65 gel released rhBMP-2 the fastest. Sponges and pads of Hyaff 1 l and Hyaff 11p80 retained less rhBMP-2 than Hyaff 11/PEG or Hyaff 11p80 gel, but more than Hyaff 11p65.
Addition of TCP to Hyaff 11 gel increased rhBMP-2 retention. The release profile in all carriers exhibited moderate to rapid burst release followed by a slow, sustained release of rhBMP-2. All Hyaff 1 l and Hyaff 11p80 gel formulations retained rhBMP-2 well (> 50 % remaining after 14 days) except Hyaff 11p65.
RAT ECTOPIC ASSAY
Hyaff 11 based gels (200 ul/site, n = 6) with 0.1 mg/ml rhBMP-2 were injected subcutaneously (ventral thorax) or injected intramuscularly (quadriceps) in 3-4 week old male Long Evans rats. Rats were sacrificed after 2 weeks and bone formation in the explants analyzed histologically using Goldners's trichrome stain. Bone scores (0= no bone, 5=100%
bone) were assigned based on histomorphometry. Total bone (mm3) was calculated using explant size and bone score. Radiographs of explants were also taken.
All Hyaff 11 based gels formed significant ectopic bone in the rat model (Table 1) in the presence of rhBMP-2, although differences in bone formation existed between carrier types as confirmed by radiographs and histology. Hyaff 11p65 at varying doses (0.1-1.5 mg/mL) of rhBMP-2 exhibited a dose dependent increase in bone formation (and bone score) but was inconsistent in explant size which yielded less total bone (0.1 mg/mL rhBMP-2 data shown).
Hyaff 11 p80 explants were large but had a lower bone score, while Hyaff 11 showed good bone score and total bone. Hyaff 11/PEG and Hyaff 11/sodium bicarbonate radiographically showed equivalent radioopacity as those of Hyaff 11 and Hyaff 11p80.
Histologically, both Hyaff 11 and Hyaff 11p80 carriers showed residual remaining matrix due to their slow degradation rates, although Hyaff 11p6~ completely degraded by 2 weeks. Bone formed within pores, shown by mineralizing osteoblasts as well as through a cartilage intermediate.
Addition of TCP to Hyaff l l gel with or without pore formers also showed comparableradiographic evidence of bone formation as those of other Hyaff based gels.
Table 1. Histomorphometry results of rat ectopic bone formation assay.
Group Bone score Total (mm3) bone SQ IM S IM
Hyaff 11p652.70 3.88 79 172 1.40 1.65 20 (33) Hyaff 11p801.83 1.83 140 314 0.68) 0.68 (76) (179) Hyaff 11 2.50 3.25 228 219 1.00) 0.96 132 (223 IN VIVO BIODISTRIBUTION
Retention of rhBMP-2 within each carrier was analyzed in vivo using a rabbit ulna fracture model. Bilateral 0.5 mm osteotomy defects were created in the ulna of New Zealand White rabbits and 150 uL rhBMP-2/carrier injected into the defect (n=8/group).
Gels were loaded with 40 uCi''SI labeled rhBMP-2 and 0.67 mg/ml unlabeled rhBMP-2.
Amount of radioactivity retained at the fracture site was measured by gamma scintigraphy as a function of time.
In vivo biodistribution of rhBMP-2 from Hyaff 11/PEG gel in the rabbit ulna fracture model showed better retention of rhBMP-2 than absorbable collagen sponge (ACS) and buffer carrier (MFR-842) (Fig. 2). Hyaff 11/PEG retained approximately 40 % rhBMP-2 after 7 days. Hyaff 11p65 gel showed poorer retention of rhBMP-2 than Hyaff 11/PEG
gel, but displayed comparable fracture callus radiographically.
IN VITRO RELEASE KINETICS
rhBMP-2 was radiolabeled with'ZSI using the Iodogen method (Pierce) and used as a tracer for 0.1 mg/ml rhBMP-2 delivered in 100 uL Hyaff 11 gel t TCP and Hyaff 11 p65 gel ~
TCP (n=4).'ZSI-rhBMP-2 loaded samples (50,000 cpm/sample) were incubated in 1 mL fetal calf serum (Hyclone) at 37°C on a shaker, and radioactivity of the carrier measured up to I 4 days using a gamma counter. .Fresh serum was replaced after 1, 3, 7, and 14 days.
Addition of TCP enhanced retention of rhBMP-2 over the course of 2 weeks in both Hyaff 11 and Hyaff 11p65 gels (Figure 3). Hyaff 11/TCP retained the most rhBMP-2, followed by Hyaff 11, Hyaff 11p65/TCP, and Hyaff 11p65. Hyaff 11 retained more rhBMP-2 than Hyaff 11p65 due to its hydrophobicity and insolubility. Preadsorbing rhBMP-2 on TCP
increased rhBMP-2 retention in Hyaff 11 gel, as opposed to mixing rhBMP-2 into the Hyaff 11 phase. Preadsorbing or mixing rhBMP-2 into either TCP or Hyaff 11p65 phase resulted in similar rhBMP-2 retention, both of which were greater than Hyaff 11p65 without TCP.
IN VIVO BIODISTRIBUTION AND EFFICACY
Retention of rhBMP-2 within Hyaff 11/TCP and Hyaff 11p65/TCP was analyzed in vivo using a rabbit ulna fracture model. Bilateral 0.5 mm osteotomy defects were created in the ulna of New Zealand White rabbits (n=3/carrier) and 150 uL Garner or buffer (0.67 mg/mL rhBMP-2) injected around the defect. 20 uCi'ZSI-rhBMP-2 was used as a tracer.
Amount of radioactivity left within each carrier at the fracture site was measured by gamma scintigraphy over the course of several weeks and in vivo rhBMP-2 retention calculated over time. Fracture repair efficacy was analyzed in these rabbits (n=8) by torsional biomechanical testing after a 4 week sacrifice to obtain maximum torque. Contralateral limbs served as surgical controls.
In vivo retention of rhBMP-2 at the rabbit ulna fracture site showed a similar pattern as that of the in vitro study (Figure 4). Hyaff 11/TCP gel (rhBMP-2 adsorbed to TCP phase first) exhibited the greatest retention (40% remaining after 4 weeks) followed by Hyaff 11p65/TCP gel (rhBMP-2 undetectable at 14 days) and buffer (undetectable at 7 days).
rhBMP-2 accelerated fracture healing when delivered in Hyaff 11p65/TCP or Hyaff 11p65 gel. Maximum torque (N-m) for Hyaff 11p65/TCP and Hyaff 11p65 were significantly greater than their contralateral surgical controls (85.6% and 96.9%, respectively) but not statistically different from each other (Table 1 ).
Table 1. Maximum torque (N-m) of rabbit ulna defects Carrier r h B M Control P value P65 0.571 0.290 0.0001 ~ 0.225 ~ 0.158 P65/TCP 0.475 ~ 0.197 0.256 ~ 0.087 0.0091 The foregoing descriptions detail presently perferred embodiments of the present invention. Numerous modifications and variations in practice thereof are expected to occur to those skilled in the art upon consideration of these descriptions. Those modifications are believed to be encompassed within the claims appended hereto.
A biodegradable matrix of porous particles for delivery of an osteogenic protein designated as OP is disclosed in Kuber A. Sampath, U.S. 5,108,753.
Brekke et al., United States Patents 4,186,448 and 5,133,755 describe methods of forming highly porous biodegradable materials composed of polymers of lactic acid ("OPLA") Okada et al., US 4,652,441, US 4,711,782, US 4,917,893 and US 5,061,492 and Yamamoto et al., US 4,954,298 disclose a prolonged-release microcapsule comprising a polypeptide drug and a drug-retaining substance encapsulated in an inner aqueous layer surrounded by a polymer wall substance in an outer oil layer.
Yamazaki et al., Clin. Orthop. and Related Research, 234:240-249 (1988) disclose the use of implants comprising 1 mg of bone morphogenetic protein purified from bone and S mg of Plaster of Paris. United States Patent 4,645,503 discloses composites of hydroxyapatite and Plaster of Paris as bone implant materials.
Collagen matrices have also been used as delivery vehicles for osteogenic proteins (see e.g. Jeffries, U.S. 4,394,370).
SUMMARY OF THE INVENTION
The present invention provides injectable formulations for delivery of osteogenic proteins. In one embodiment the composition comprises the osteogenic protein and hyaluronic acid esters. In another embodiment, the composition may further include tricalcium phosphate. The injectable formulations of the invention allows for closed fracture repair and other skeletal tissue without an open reduction procedure as is necessary with implantable devices.
The present invention further provides methods for preparing injectable gels or pastes useful as a carrier for osteogenic proteins by transforming various non-woven pads and sponges of hyaluronic acid benzyl ester into injectable gel or paste formulations by hydration or solvent addition. In another embodiment, the invention comprises compositions comprising the transformed injectable gel or paste formulations.
The methods and compositions of the present invention are useful for the preparation of formulations of osteoinductive proteins which can be used, among other uses, to promote the formation of cartilage and/or bone, for repair of tissue damage and fractures. The invention further provides methods for treating patients in need of cartilage and/or bone repair and/or growth.
_J_ BRIEF DESCRIPTION OF THE DRAWINGS
Figure 1 sets forth in vitro release kinetics of'z5I-rhBMP-2 in Hyaff gels.
Figure 2 sets forth in vivo retention of lzsl_rhBMP-2 in Hyaff 11/PEG, ACS, and buffer.
Figure 3 sets forth in vitro release kinetics of'z5I-rhBMP-2 in Hyaff gels/TCP.
S Figure 4 sets forth in vivo biodistribution of'zsI_rhBMP-2.
DETAILED DESCRIPTION OF THE INVENTION
The invention provides injectable formulations for delivery of osteogenic proteins.
The compositions comprise an injectable formulation of hyaluronic acid esters and osteogenic protein. The present invention further provides processes for preparing injectable gel or paste formulations by transforming various non-woven pads and sponges of hyaluronic acid benzyl ester by hydration or solvent addition yielding gels with in vivo residence times from days to up to several months. Total or partial esters of hyaluronic acid are described in US 5,336,767.
Partial esters of Hyaff solids are transformed into gels using aqueous buffer or organic solvents (such as N-methyl pyrrolidinone, dimethyl sulfoxide, etc), while complete esters of Hyaff solids are transformed into gels using organic solvents. In other embodiments pore formers may be introduced to the solublized carriers to increase porosity. The addition of pore formers would allow in situ pore formation after injection in vivo by solubilization of pore former and precipitation/phase inversion of Garner. Suitable liquid pore formers include polyethylene glycol or PEG at 10 - 90 % volume per volume ratios) and solid pore formers (such as sodium bicarbonate, sodium chloride, citric acid, sucrose, etc., at 1:1 - 21:1 pore former:Hyaff weight per weight ratios) to increase porosity. The gel/paste can also contain TCP (tri-calcium phosphate) particles as a mineral component for example, at 0.1 - 100 weight per volume range.
The amount , type and size of the pore forming agent is optimized to leave voids sufficient for cell ingrowth into injectable gel when pore forming agent and solvent are extracted from the carrier in vivo by solubilization of pore forming agent and precipitation/phase inversion of carrier in situ.
The osteogenic proteins useful with the injectable carriers made in accordance with the subject invention are well known to those skilled in the art and include those discussed above. The preferred osteogenic proteins for use herein are those of the BMP
class identified as BMP-1 through BMP-12 in US 4,877,864; US 5,013,649; WO 90/11366 published October 4, 1990; WO 91/18098 published November 28, 1991; WO 93/00432, published January 7, 1993; United States Serial Numbers 08/247,908 and 08/247,904, both filed May 20, 1994; and United States Serial Number 08/217,780, filed on March 25, 1994.
The disclosure of the above publications are hereby incorporated by reference. The most preferred is BMP-2, the full length cDNA sequence of which is described in detail in the '649 patent.
Of course, combinations of two or more of such osteogenic proteins may be used, as may fragments of such proteins that also exhibit osteogenic activity. Such osteogenic proteins are known to be homodimeric species, but also exhibit activity as mixed heterodimers.
Heterodimeric forms of osteogenic proteins may also be used in the practice of the subject invention. BMP heterodimers are described in W093/09229, the disclosure of which is hereby incorporated by reference. Recombinant proteins are preferred over naturally occurnng isolated proteins. The amount of osteogenic protein useful herein is that amount effective to stimulate increased osteogenic activity of infiltrating progenitor cells, and will depend upon the size and nature of defect being treated as well as the carrier being employed.
The formulations may be injected for example into tendons, damaged cartilage tissue, ligaments, and/or their attachment sites to bones. Injectable formulations may also fmd application to other bone sites such as bone cysts, bone defects, intraosseous sites and closed fractures.
The dosage regimen will be determined by the clinical indication being addressed, as well as by various patient variables (e.g. weight, age, sex) and clinical presentation (e.g.
extent of injury, site of injury, etc.). In general, the dosage of osteogenic protein will'be in the range of from about 0.1 to 4 mg/ml.
The injectable osteogenic protein formulations may be provided to the clinic as a single formulation, or the formulation may be provided as a multicomponent kit wherein, e.g.
the osteogenic protein is provided in one vial and the injectable hyaluronic paste is provided separately.
The compositions of the subject invention allow therapeutically effective amounts of osteoinductive protein to be delivered to an injury site where cartilage and/or bone formation is desired. The formulations may be used as a substitute for autologous bone graft in fresh and non-union fractures, spinal fusions, and bone defect repair in the orthopaedic field; in _j_ cranio/maxillofacial reconstructions; for prosthesis integration, especially as a surface coating to improve fixation of prosthetic implants such as hydroxyapatite coated prostheses; in osteomyelitis for bone regeneration; and in the dental field for augmentation of the alveolar ridge and periodontal defects and tooth extraction sockets. The methods and formulations of the present invention may be useful in the treatment and/or prevention of osteoporosis, or the treatment of osteoporotic or osteopenic bone. In another embodiment, formulations of the present invention may be used in the process known as distraction osteogenesis. When used to treat osteomyelitis or for bone repair with minimal infection, the osteogenic protein may be used in combination with porous microparticles and antibiotics, with the addition of protein sequestering agents such as alginate, cellulosics, especially carboxymethylcellulose, diluted using aqueous glycerol. The antibiotic is selected for its ability to decrease infection while having minimal adverse effects on bone formation. Preferred antibiotics for use in the devices of the present invention include vancomycin and gentamycin. The antibiotic may be in any pharmaceutically acceptable form, such as vancomycin HCl or gentamycin sulfate. The antibiotic is preferably present in a concentration of from about 0.1 mg/mL to about 10.0 mg/mL. The traditional preparation of formulations in pharmaceutically acceptable form (i.e.
pyrogen free, appropriate pH and isotonicity, sterility, etc.) is well within the skill in the art and is applicable to the formulations of the invention.
Hyaluronic derivitive compositions of the invention prepared by hydration or solvent addition of insoluble or partially soluble non- woven pads or sponges may also be ultilized in combination with other drugs, growth factors, peptides, proteins, cytokines, oligonucleeotides antisense oligonucleotides, DNA and polymers. These compounds may be added by mixing them with the carriers. Or by covalent attachment to the polymer carriers. The hyaluronic derivitive compositions may also be used with DNA encoding for BMPs and cells transduced or transfected with genes encoding BMP proteins.
The following examples are illustrative of the present invention and are not limiting in any manner. Modifications, variations and minor enhancements are contemplated and are within the present invention.
PREPARATION OF INJECTABLE HYALURONIC ACID ESTERS
The starting Hyaff hyaluronic acid (Fidia Advanced Biopolymers, Abano Terme, Italy) materials are solids such as non-woven pads, felts, sheets, powders, sponges, and microspheres. The Hyaff materials are esters of hyaluronic acid exhibiting various ester moities (e.g., benzyl, ethyl,propyl pentyl or larger molecules such as hydrocortisone or methyl prednislone, etc.) as well as various degrees of esterification (i.e., partial esters or complete esters). Partial esters of Hyaff are designated by percent esterfication ranging from 50-99 (e.g., Hyaff 11p65, Hyaff 11p80, etc.), while complete esters are 100 % esters of hyaluronic acid (e.g., Hyaff 11).
Hyaff gel classification used in supporting data is as follows and is followed by examples of select formulations:
- Hyaff 11 gel: Hyaff 11 non-woven pad transformed into gel with organic solvent to yield 10 % solids - Hyaff 11/bicarbonate gel: Hyaff 11 gel mixed with sodium bicarbonate as pore former at 15:1 (w/w) bicarbonate to Hyaff 11 - Hyaff 11/PEG gel: Hyaff 11 gel mixed with polyethylene glycol(200mw) as pore former at 33 - 50 % (v/v) range - Hyaff 11/TCP gel: Hyaff 11 gel mixed with 30 % w/v TCP
- Hyaff 11/bicarbonate/TCP gel: Hyaff 11/bicarbonate gel mixed with 30 % w/v TCP
- Hyaff 11/PEG/TCP gel: Hyaff 11/PEG gel mixed with 30 % w/v TCP
- Hyaff 11p80 gel: Hyaff 11p80 non-woven pad transformed into gel with organic solvent to yield 5 % solids - Hyaff 11p65 gel: Hyaff 11p65 non-woven pad hydrated with aqueous buffer to yield 6 -15 % solids - Hyaff 11p65/TCP gel: Hyaff 11p65 gel mixed with 30 % w/v TCP
Hyaff 11p65 non-woven pads were hydrated with glutamic acid buffer (pH 4.5) containing rhBMP-2 (0.1 mg/mL final cone) to yield either 6 % - 15 % solids (w/v) and mixed thoroughly to form a paste. Hyaff 11p80 and Hyaff 11 non-woven pads were solubilized in N-methyl-pyrrolidinone (NMP) or dimethyl sulfoxide(DMSO)to yield a 1 - 30 _7_ w/v solution. These solutions were then mixed with either rhBMP-2-containing buffer (10 v/v, 0.1 mg/mL rhBMP-2), or lyophilized rhBMP-2 (0.1 mg/mL) followed by the addition of various pore formers (polyethylene glycol, sodium bicarbonate, sucrose, NaCI, citric acid)and tricalcium phosphate (TCP). Particle size of solid pore formers and TCP used was <
600um, preferably <200umLiquid pore formers such as PEG(200mw) were mixed at 10-90%
v/v ratios, and solid pore formers were mixed at 9:1 - 21:1 (w/w) pore former to carrier ratios.
TCP was mixed at 0.1-30% (w/v). TCP (45-125 micron particle size) was mixed thoroughly into rhBMP-2/Hyaff 11 or rhBMP-2/Hyaff 11p65 gel at 30% (w/v). Separately, rhBMP-2 was adsorbed onto TCP first, followed by mixing with Hyaff 11 or Hyaff 11p65 gel.Formulations were chosen based on injectability through an 18 g needle.
Microstructure was characterized by scanning electron microscopy (SEM).
SEMS revealed varying degrees of pore structure and porosity. Hyaff 11p65 6%
gel exhibited longer fibers than the 15% formulation; with both displaying a high level of porosity. Both Hyaff 11 and Hyaff 11p80 gels showed minimal pore structure and porosity, whereas those carriers with pore formers displayed a high level of porosity.
Pore formers and/
or additives that yielded injectable mixtures were PEG, sodium bicarbonate and TCP.
IN VITRO RELEASE KINETICS
rhBMP-2 was radiolabeled with'zsI using the Iodogen method (Pierce) and used as a tracer for 0.1 mg/ml rhBMP-2 delivered in 100 u1 Hyaff=11p65 gel, Hyaff 11p80 gel, Hyaff 11 gel and Hyaff 11lPEG (n=4). 'z5I-rhBMP-2 loaded samples (50,000 cpm/sample) were incubated in 1 ml fetal calf serum (Hyclone) at 37°C on a shaker, and radioactivity of the Garner measured up to 14 days using a gamma counter. Fresh serum was replaced after each time point. 'z5I-rhBMP-2 release from injectable formulations were compared to those of implantable sponges and pads of Hyaff 11 and Hyaff 11p80.
Auto cross-linked polysaccharide form of derivitized hyaluronic acid, ACP gel, is used for the in vitro release study and the rat ectopic assay. For the in vitro release study, 2 ml ACP gel is mixed with 1.53 mg rhBMP-2 cake (which corresponds to 0.2 mg actual rhBMP-2 at 8 mg rhBMP-2 per 61 mg cake weight) and'z5I-rhBMP-2 (100 ml total, 20 mCi/200 ml gel) and drawn up into 1 ml syringes resulting in approximately 10 % gel dilution. ACP
gel for the rat _g_ ectopic study does not contain the tracer but is diluted with MRF-00906 buffer. 200 ml injections are performed using a 22 gauge needle. The final concentration of rhBMP-2 will be 0.1 mg/ml, or 20 mg per 200 ml injection. The final concentration of '251-rhBMP-2 will be approximately 20 mCi per 200 ml injection. The ACP gel will be injected at room temperature.
In vitro release kinetics showed greatest retention of rhBMP-2 over the 2 weeks in the Hyaff 11/PEG gel followed by Hyaff 11p80 gel and Hyaff 11 gel (Fig. 1). Hyaff 11p65 gel released rhBMP-2 the fastest. Sponges and pads of Hyaff 1 l and Hyaff 11p80 retained less rhBMP-2 than Hyaff 11/PEG or Hyaff 11p80 gel, but more than Hyaff 11p65.
Addition of TCP to Hyaff 11 gel increased rhBMP-2 retention. The release profile in all carriers exhibited moderate to rapid burst release followed by a slow, sustained release of rhBMP-2. All Hyaff 1 l and Hyaff 11p80 gel formulations retained rhBMP-2 well (> 50 % remaining after 14 days) except Hyaff 11p65.
RAT ECTOPIC ASSAY
Hyaff 11 based gels (200 ul/site, n = 6) with 0.1 mg/ml rhBMP-2 were injected subcutaneously (ventral thorax) or injected intramuscularly (quadriceps) in 3-4 week old male Long Evans rats. Rats were sacrificed after 2 weeks and bone formation in the explants analyzed histologically using Goldners's trichrome stain. Bone scores (0= no bone, 5=100%
bone) were assigned based on histomorphometry. Total bone (mm3) was calculated using explant size and bone score. Radiographs of explants were also taken.
All Hyaff 11 based gels formed significant ectopic bone in the rat model (Table 1) in the presence of rhBMP-2, although differences in bone formation existed between carrier types as confirmed by radiographs and histology. Hyaff 11p65 at varying doses (0.1-1.5 mg/mL) of rhBMP-2 exhibited a dose dependent increase in bone formation (and bone score) but was inconsistent in explant size which yielded less total bone (0.1 mg/mL rhBMP-2 data shown).
Hyaff 11 p80 explants were large but had a lower bone score, while Hyaff 11 showed good bone score and total bone. Hyaff 11/PEG and Hyaff 11/sodium bicarbonate radiographically showed equivalent radioopacity as those of Hyaff 11 and Hyaff 11p80.
Histologically, both Hyaff 11 and Hyaff 11p80 carriers showed residual remaining matrix due to their slow degradation rates, although Hyaff 11p6~ completely degraded by 2 weeks. Bone formed within pores, shown by mineralizing osteoblasts as well as through a cartilage intermediate.
Addition of TCP to Hyaff l l gel with or without pore formers also showed comparableradiographic evidence of bone formation as those of other Hyaff based gels.
Table 1. Histomorphometry results of rat ectopic bone formation assay.
Group Bone score Total (mm3) bone SQ IM S IM
Hyaff 11p652.70 3.88 79 172 1.40 1.65 20 (33) Hyaff 11p801.83 1.83 140 314 0.68) 0.68 (76) (179) Hyaff 11 2.50 3.25 228 219 1.00) 0.96 132 (223 IN VIVO BIODISTRIBUTION
Retention of rhBMP-2 within each carrier was analyzed in vivo using a rabbit ulna fracture model. Bilateral 0.5 mm osteotomy defects were created in the ulna of New Zealand White rabbits and 150 uL rhBMP-2/carrier injected into the defect (n=8/group).
Gels were loaded with 40 uCi''SI labeled rhBMP-2 and 0.67 mg/ml unlabeled rhBMP-2.
Amount of radioactivity retained at the fracture site was measured by gamma scintigraphy as a function of time.
In vivo biodistribution of rhBMP-2 from Hyaff 11/PEG gel in the rabbit ulna fracture model showed better retention of rhBMP-2 than absorbable collagen sponge (ACS) and buffer carrier (MFR-842) (Fig. 2). Hyaff 11/PEG retained approximately 40 % rhBMP-2 after 7 days. Hyaff 11p65 gel showed poorer retention of rhBMP-2 than Hyaff 11/PEG
gel, but displayed comparable fracture callus radiographically.
IN VITRO RELEASE KINETICS
rhBMP-2 was radiolabeled with'ZSI using the Iodogen method (Pierce) and used as a tracer for 0.1 mg/ml rhBMP-2 delivered in 100 uL Hyaff 11 gel t TCP and Hyaff 11 p65 gel ~
TCP (n=4).'ZSI-rhBMP-2 loaded samples (50,000 cpm/sample) were incubated in 1 mL fetal calf serum (Hyclone) at 37°C on a shaker, and radioactivity of the carrier measured up to I 4 days using a gamma counter. .Fresh serum was replaced after 1, 3, 7, and 14 days.
Addition of TCP enhanced retention of rhBMP-2 over the course of 2 weeks in both Hyaff 11 and Hyaff 11p65 gels (Figure 3). Hyaff 11/TCP retained the most rhBMP-2, followed by Hyaff 11, Hyaff 11p65/TCP, and Hyaff 11p65. Hyaff 11 retained more rhBMP-2 than Hyaff 11p65 due to its hydrophobicity and insolubility. Preadsorbing rhBMP-2 on TCP
increased rhBMP-2 retention in Hyaff 11 gel, as opposed to mixing rhBMP-2 into the Hyaff 11 phase. Preadsorbing or mixing rhBMP-2 into either TCP or Hyaff 11p65 phase resulted in similar rhBMP-2 retention, both of which were greater than Hyaff 11p65 without TCP.
IN VIVO BIODISTRIBUTION AND EFFICACY
Retention of rhBMP-2 within Hyaff 11/TCP and Hyaff 11p65/TCP was analyzed in vivo using a rabbit ulna fracture model. Bilateral 0.5 mm osteotomy defects were created in the ulna of New Zealand White rabbits (n=3/carrier) and 150 uL Garner or buffer (0.67 mg/mL rhBMP-2) injected around the defect. 20 uCi'ZSI-rhBMP-2 was used as a tracer.
Amount of radioactivity left within each carrier at the fracture site was measured by gamma scintigraphy over the course of several weeks and in vivo rhBMP-2 retention calculated over time. Fracture repair efficacy was analyzed in these rabbits (n=8) by torsional biomechanical testing after a 4 week sacrifice to obtain maximum torque. Contralateral limbs served as surgical controls.
In vivo retention of rhBMP-2 at the rabbit ulna fracture site showed a similar pattern as that of the in vitro study (Figure 4). Hyaff 11/TCP gel (rhBMP-2 adsorbed to TCP phase first) exhibited the greatest retention (40% remaining after 4 weeks) followed by Hyaff 11p65/TCP gel (rhBMP-2 undetectable at 14 days) and buffer (undetectable at 7 days).
rhBMP-2 accelerated fracture healing when delivered in Hyaff 11p65/TCP or Hyaff 11p65 gel. Maximum torque (N-m) for Hyaff 11p65/TCP and Hyaff 11p65 were significantly greater than their contralateral surgical controls (85.6% and 96.9%, respectively) but not statistically different from each other (Table 1 ).
Table 1. Maximum torque (N-m) of rabbit ulna defects Carrier r h B M Control P value P65 0.571 0.290 0.0001 ~ 0.225 ~ 0.158 P65/TCP 0.475 ~ 0.197 0.256 ~ 0.087 0.0091 The foregoing descriptions detail presently perferred embodiments of the present invention. Numerous modifications and variations in practice thereof are expected to occur to those skilled in the art upon consideration of these descriptions. Those modifications are believed to be encompassed within the claims appended hereto.
Claims (10)
1. A composition for injectable delivery of osteogenic proteins comprising a pharmaceutically acceptable admixture comprising (a) an osteogenic protein; and (b) injectable hyaluronic acid derivitives.
2. The composition of claim 1 further comprising tricalcium phosphate.
3. The composition of claim 1 further comprising pore formers.
4. The composition of claim 1 wherein the osteogenic protein is selected from the group consisting of members of the BMP family.
5. The composition of claim 4 wherein the osteogenic protein is BMP-2.
6. The composition of claim 4 wherein the osteogenic protein is OP-1.
7. A composition for injectable delivery of osteogenic proteins comprising a pharmaceutically acceptable admixture comprising (a) BMP-2;
(b) an injectable hyaluronic acid derivitive; and (c) Tricalcium phosphate.
(b) an injectable hyaluronic acid derivitive; and (c) Tricalcium phosphate.
8. A composition providing an injectable matrix for osteogenic proteins comprising a hyaluronic acid derivitive.
9. A method for making an injectable hyaluronic acid derivitive comprising transforming insoluble or partially solublenon-woven pads or sponges of hyaluronic acid benzyl esters by hydration or solvent addition.
10. The method of claim 9 further comprising the introduction of pore formers to the solubilized carriers.
Applications Claiming Priority (5)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US15967499P | 1999-10-15 | 1999-10-15 | |
US60/159,674 | 1999-10-15 | ||
US18558700P | 2000-02-28 | 2000-02-28 | |
US60/185,587 | 2000-02-28 | ||
PCT/US2000/028468 WO2001028602A1 (en) | 1999-10-15 | 2000-10-13 | Formulations of hyaluronic acid for delivery of osteogenic proteins |
Publications (1)
Publication Number | Publication Date |
---|---|
CA2386408A1 true CA2386408A1 (en) | 2001-04-26 |
Family
ID=26856175
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CA002386408A Abandoned CA2386408A1 (en) | 1999-10-15 | 2000-10-13 | Formulations of hyaluronic acid for delivery of osteogenic proteins |
Country Status (11)
Country | Link |
---|---|
US (2) | US7189392B1 (en) |
EP (2) | EP1223990B1 (en) |
JP (1) | JP4703926B2 (en) |
AT (1) | ATE271886T1 (en) |
AU (2) | AU774427B2 (en) |
CA (1) | CA2386408A1 (en) |
DE (1) | DE60012557T2 (en) |
DK (1) | DK1223990T3 (en) |
ES (1) | ES2225241T3 (en) |
PT (1) | PT1223990E (en) |
WO (1) | WO2001028602A1 (en) |
Families Citing this family (32)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20070026437A1 (en) * | 1988-04-08 | 2007-02-01 | Genetics Institute, L.L.C. | Novel BMP products |
US7582311B1 (en) * | 1999-10-15 | 2009-09-01 | Genentech, Inc. | Injection vehicle for polymer-based formulations |
PT1223990E (en) * | 1999-10-15 | 2004-12-31 | Fidia Advanced Biopolymers Srl | HYALURONIC ACID FORMULATIONS FOR ADMINISTRATION OF OSTEOGENIC PROTEINS |
US20020114795A1 (en) | 2000-12-22 | 2002-08-22 | Thorne Kevin J. | Composition and process for bone growth and repair |
TWI267378B (en) * | 2001-06-08 | 2006-12-01 | Wyeth Corp | Calcium phosphate delivery vehicles for osteoinductive proteins |
AU2007203555B2 (en) * | 2001-06-08 | 2009-01-08 | Etex Corporation | Calcium Phosphate Delivery Vehicles for Osteoinductive Proteins |
CN100425285C (en) | 2001-11-19 | 2008-10-15 | Scil技术股份有限公司 | Device having osteoinductive and osteoconductive properties |
EP1519744A4 (en) | 2002-05-17 | 2007-10-03 | Wyeth Corp | Injectable solid hyaluronic acid carriers for delivery of osteogenic proteins |
EP1539261B1 (en) | 2002-09-10 | 2006-04-12 | Scil Technology GmbH | Metal implant coated under reduced oxygen concentration with osteoinductive protein |
JP5392981B2 (en) * | 2003-09-12 | 2014-01-22 | ワイス・エルエルシー | Injectable calcium phosphate solid rods and pastes for delivery of bone morphogenetic proteins |
JP4944010B2 (en) | 2004-03-10 | 2012-05-30 | サイル テクノロジー ゲーエムベーハー | Coated implant, its manufacture and use |
ES2246695B1 (en) | 2004-04-29 | 2007-05-01 | Instituto Cientifico Y Tecnologico De Navarra, S.A. | STIMULATING COMPOSITION OF THE IMMUNE RESPONSE THAT INCLUDES NANOPARTICLES BASED ON A COPYLIMER OF METHYL VINYL ETER AND MALEIC ANHYDRIDE. |
ES2246694B1 (en) | 2004-04-29 | 2007-05-01 | Instituto Cientifico Y Tecnologico De Navarra, S.A. | PEGILATED NANOPARTICLES. |
ITPD20040312A1 (en) * | 2004-12-15 | 2005-03-15 | Fidia Advanced Biopolymers Srl | PROSTHESIS AND SUPPORT FOR REPLACEMENT, REPAIR, REGENERATION OF THE MENISCUS |
US8048857B2 (en) * | 2006-12-19 | 2011-11-01 | Warsaw Orthopedic, Inc. | Flowable carrier compositions and methods of use |
US7718616B2 (en) * | 2006-12-21 | 2010-05-18 | Zimmer Orthobiologics, Inc. | Bone growth particles and osteoinductive composition thereof |
FR2919188B1 (en) * | 2007-07-27 | 2010-02-26 | Proteins & Peptides Man | COMPLEXES BETWEEN AN AMPHIPHILIC POLYMER AND A OSTEOGENIC PROTEIN BELONGING TO THE BMPS FAMILY |
JP2011503047A (en) | 2007-11-13 | 2011-01-27 | バイオ−テクノロジー ゼネラル(イスラエル)リミテッド | Dilution filtration sterilization process for viscoelastic biopolymers |
US9320761B2 (en) | 2008-12-18 | 2016-04-26 | Vivex Biomedical, Inc. | Bone induction system and methods |
US8613938B2 (en) | 2010-11-15 | 2013-12-24 | Zimmer Orthobiologics, Inc. | Bone void fillers |
US8455436B2 (en) | 2010-12-28 | 2013-06-04 | Depuy Mitek, Llc | Compositions and methods for treating joints |
US8524662B2 (en) | 2010-12-28 | 2013-09-03 | Depuy Mitek, Llc | Compositions and methods for treating joints |
US8398611B2 (en) | 2010-12-28 | 2013-03-19 | Depuy Mitek, Inc. | Compositions and methods for treating joints |
US9265830B2 (en) | 2011-04-20 | 2016-02-23 | Warsaw Orthopedic, Inc. | Implantable compositions and methods for preparing the same |
US8623839B2 (en) | 2011-06-30 | 2014-01-07 | Depuy Mitek, Llc | Compositions and methods for stabilized polysaccharide formulations |
RU2533017C1 (en) * | 2013-06-25 | 2014-11-20 | Федеральное государственное бюджетное учреждение "Саратовский научно-исследовательский институт травматологии и ортопедии" Министерства здравоохранения Российской Федерации (ФГБУ "СарНИИТО" Минздрава России) | Method of treating ganglia of long bones |
US9682099B2 (en) | 2015-01-20 | 2017-06-20 | DePuy Synthes Products, Inc. | Compositions and methods for treating joints |
EP3250251A1 (en) * | 2015-01-28 | 2017-12-06 | Allergan, Inc. | Joint fat pad formulations, and methods of use thereof |
CA2986702C (en) | 2015-05-21 | 2023-04-04 | David Wang | Modified demineralized cortical bone fibers |
US10675330B2 (en) * | 2018-03-21 | 2020-06-09 | Warsaw Orthopedic, Inc. | Injectable bone morphogenetic protein |
CN108324994B (en) * | 2018-04-26 | 2020-12-01 | 海口市人民医院(中南大学湘雅医学院附属海口医院) | Preparation method of artificial bone of pearl powder |
US11324806B2 (en) | 2018-10-19 | 2022-05-10 | Warsaw Orthopedic, Inc. | Sustained delivery of a growth differentiation factor |
Family Cites Families (142)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US2465357A (en) | 1944-08-14 | 1949-03-29 | Upjohn Co | Therapeutic sponge and method of making |
CH563767A5 (en) * | 1973-01-30 | 1975-07-15 | Pheulpin Jean | |
US4468464A (en) | 1974-11-04 | 1984-08-28 | The Board Of Trustees Of The Leland Stanford Junior University | Biologically functional molecular chimeras |
US4186448A (en) | 1976-04-16 | 1980-02-05 | Brekke John H | Device and method for treating and healing a newly created bone void |
DE2657370C2 (en) | 1976-12-17 | 1982-11-11 | Hans Dr.med. Dr.med.dent. 8000 München Scheicher | Means for covering and / or filling in bone defects |
DE2732848A1 (en) | 1977-07-18 | 1979-02-08 | Schering Ag | DIURETHANE, HERBICIDAL AGENTS CONTAINING THESE COMPOUNDS AND THE PROCESS FOR THEIR PRODUCTION |
US4399216A (en) | 1980-02-25 | 1983-08-16 | The Trustees Of Columbia University | Processes for inserting DNA into eucaryotic cells and for producing proteinaceous materials |
US4761471A (en) | 1980-08-04 | 1988-08-02 | The Regents Of The University Of California | Bone morphogenetic protein composition |
US4789732A (en) | 1980-08-04 | 1988-12-06 | Regents Of The University Of California | Bone morphogenetic protein composition |
US4455256A (en) | 1981-05-05 | 1984-06-19 | The Regents Of The University Of California | Bone morphogenetic protein |
US4294753A (en) | 1980-08-04 | 1981-10-13 | The Regents Of The University Of California | Bone morphogenetic protein process |
US4619989A (en) | 1981-05-05 | 1986-10-28 | The Regents Of The University Of Cal. | Bone morphogenetic protein composition |
US4419446A (en) | 1980-12-31 | 1983-12-06 | The United States Of America As Represented By The Department Of Health And Human Services | Recombinant DNA process utilizing a papilloma virus DNA as a vector |
IE52535B1 (en) | 1981-02-16 | 1987-12-09 | Ici Plc | Continuous release pharmaceutical compositions |
US4727028A (en) | 1981-06-22 | 1988-02-23 | Eli Lilly And Company | Recombinant DNA cloning vectors and the eukaryotic and prokaryotic transformants thereof |
US4472840A (en) | 1981-09-21 | 1984-09-25 | Jefferies Steven R | Method of inducing osseous formation by implanting bone graft material |
US4394370A (en) | 1981-09-21 | 1983-07-19 | Jefferies Steven R | Bone graft material for osseous defects and method of making same |
US4474181A (en) | 1982-02-18 | 1984-10-02 | Schenck Robert R | Method and apparatus for anastomosing small blood vessels |
IL68218A (en) | 1983-03-23 | 1985-12-31 | Univ Ramot | Compositions for cartilage repair comprising embryonal chondrocytes |
US4434094A (en) | 1983-04-12 | 1984-02-28 | Collagen Corporation | Partially purified osteogenic factor and process for preparing same from demineralized bone |
US4795804A (en) | 1983-08-16 | 1989-01-03 | The Regents Of The University Of California | Bone morphogenetic agents |
US4923805A (en) | 1983-11-02 | 1990-05-08 | Integrated Genetics, Inc. | Fsh |
JPS60100516A (en) | 1983-11-04 | 1985-06-04 | Takeda Chem Ind Ltd | Preparation of sustained release microcapsule |
US4703008A (en) | 1983-12-13 | 1987-10-27 | Kiren-Amgen, Inc. | DNA sequences encoding erythropoietin |
GB8334498D0 (en) | 1983-12-24 | 1984-02-01 | Beecham Group Plc | Compounds |
US4804744A (en) | 1984-01-04 | 1989-02-14 | International Genetic Engineering, Inc. | Osteogenic factors |
US4526909A (en) | 1984-01-09 | 1985-07-02 | Regents Of The University Of California | Polymethylmethacrylate delivery system for bone morphogenetic protein |
US4563489A (en) | 1984-02-10 | 1986-01-07 | University Of California | Biodegradable organic polymer delivery system for bone morphogenetic protein |
DE3587022T2 (en) | 1984-02-17 | 1993-06-17 | Genentech Inc | HUMAN TRANSFORMATION GROWTH FACTOR AND PRECURSOR OR FRAGMENT THEREOF, CELLS, DNA, VECTORS AND METHODS FOR THE PRODUCTION THEREOF, COMPOSITIONS AND PRODUCTS THAT CONTAIN THESE, AND ANTI-OXIDERS AND DIAGNOSTICS DERIVED FROM THEM. |
US4608199A (en) | 1984-03-20 | 1986-08-26 | Arnold Caplan | Bone protein purification process |
US4662884A (en) | 1984-04-25 | 1987-05-05 | University Of Utah Research Foundation | Prostheses and methods for promoting nerve regeneration |
US4596574A (en) | 1984-05-14 | 1986-06-24 | The Regents Of The University Of California | Biodegradable porous ceramic delivery system for bone morphogenetic protein |
CA1341617C (en) | 1984-06-08 | 2011-06-28 | Henry George Burger | Inhibin isolated from ovarian follicular fluid |
US4868161A (en) | 1984-06-29 | 1989-09-19 | City Of Hope | Method for promoting nerve regeneration |
US4843063A (en) | 1984-07-16 | 1989-06-27 | Collagen Corporation | Polypeptide cartilage-inducing factors found in bone |
DE3588058T3 (en) | 1984-07-16 | 2005-04-07 | Celtrix Pharmaceuticals, Inc., Palo Alto | Cartilage-inducing polypeptide factors from bone |
US4627982A (en) | 1984-07-16 | 1986-12-09 | Collagen Corporation | Partially purified bone-inducing factor |
US4769328A (en) | 1984-10-12 | 1988-09-06 | Zymogenetics Inc. | Expression of biologically active PDGF analogs in yeast |
US5187263A (en) | 1984-10-12 | 1993-02-16 | Zymogenetics, Inc. | Expression of biologically active PDGE analogs in eucaryotic cells |
US4563350A (en) | 1984-10-24 | 1986-01-07 | Collagen Corporation | Inductive collagen based bone repair preparations |
AU598455B2 (en) | 1984-12-27 | 1990-06-28 | Suntory Limited | Method for purifying an interferon |
ATE61935T1 (en) | 1985-02-07 | 1991-04-15 | Takeda Chemical Industries Ltd | PROCESS FOR PRODUCTION OF MICROCAPSULES. |
US4886747A (en) | 1985-03-22 | 1989-12-12 | Genentech, Inc. | Nucleic acid encoding TGF-β and its uses |
US4766067A (en) | 1985-05-31 | 1988-08-23 | President And Fellows Of Harvard College | Gene amplification |
US4681763A (en) | 1985-06-11 | 1987-07-21 | University Of Medicine And Dentistry Of New Jersey | Composition for stimulating bone growth |
US4851521A (en) * | 1985-07-08 | 1989-07-25 | Fidia, S.P.A. | Esters of hyaluronic acid |
US4645503A (en) | 1985-08-27 | 1987-02-24 | Orthomatrix Inc. | Moldable bone-implant material |
US5215893A (en) | 1985-10-03 | 1993-06-01 | Genentech, Inc. | Nucleic acid encoding the ba chain prodomains of inhibin and method for synthesizing polypeptides using such nucleic acid |
US4798885A (en) | 1986-02-07 | 1989-01-17 | Genentech, Inc. | Compositions of hormonally active human and porcine inhibin containing an α chain and 62 chain |
US5089396A (en) | 1985-10-03 | 1992-02-18 | Genentech, Inc. | Nucleic acid encoding β chain prodomains of inhibin and method for synthesizing polypeptides using such nucleic acid |
US5133755A (en) | 1986-01-28 | 1992-07-28 | Thm Biomedical, Inc. | Method and apparatus for diodegradable, osteogenic, bone graft substitute device |
US4737578A (en) | 1986-02-10 | 1988-04-12 | The Salk Institute For Biological Studies | Human inhibin |
US4758233A (en) * | 1986-04-22 | 1988-07-19 | N.J. Phillips TPY. Limited | Cream applicator |
NL8601328A (en) * | 1986-05-23 | 1987-12-16 | Langen Research | DEVICE FOR INJECTING MEAT WITH A MASS, PARTICULARLY PASTY MASS. |
US5106748A (en) | 1986-07-01 | 1992-04-21 | Genetics Institute, Inc. | Dna sequences encoding 5 proteins |
US5013649A (en) | 1986-07-01 | 1991-05-07 | Genetics Institute, Inc. | DNA sequences encoding osteoinductive products |
ZA874681B (en) | 1986-07-01 | 1988-04-27 | Genetics Inst | Novel osteoinductive factors |
US5187076A (en) | 1986-07-01 | 1993-02-16 | Genetics Institute, Inc. | DNA sequences encoding BMP-6 proteins |
US5108922A (en) | 1986-07-01 | 1992-04-28 | Genetics Institute, Inc. | DNA sequences encoding BMP-1 products |
US4877864A (en) | 1987-03-26 | 1989-10-31 | Genetics Institute, Inc. | Osteoinductive factors |
US5366875A (en) | 1986-07-01 | 1994-11-22 | Genetics Institute, Inc. | Methods for producing BMP-7 proteins |
US5019087A (en) | 1986-10-06 | 1991-05-28 | American Biomaterials Corporation | Nerve regeneration conduit |
IT1198449B (en) | 1986-10-13 | 1988-12-21 | F I D I Farmaceutici Italiani | ESTERS OF POLYVALENT ALCOHOLS OF HYALURONIC ACID |
US5124316A (en) | 1986-11-14 | 1992-06-23 | President And Fellows Of Harvard College | Method for periodontal regeneration |
US5001691A (en) | 1986-12-15 | 1991-03-19 | Antonov Alexandr A | High density optical storage device |
US5041538A (en) | 1987-08-28 | 1991-08-20 | The Salk Institute For Biological Studies | Mammalian follistatin |
US5202120A (en) | 1987-09-11 | 1993-04-13 | Case Western Reserve University | Methods of reducing glial scar formation and promoting axon and blood vessel growth and/or regeneration through the use of activated immature astrocytes |
US5147399A (en) | 1988-02-01 | 1992-09-15 | Dellon Arnold L | Method of treating nerve defects through use of a bioabsorbable surgical device |
US5108753A (en) | 1988-04-08 | 1992-04-28 | Creative Biomolecules | Osteogenic devices |
US5258494A (en) | 1988-04-08 | 1993-11-02 | Stryker Corporation | Osteogenic proteins |
US5011691A (en) | 1988-08-15 | 1991-04-30 | Stryker Corporation | Osteogenic devices |
US5266683A (en) | 1988-04-08 | 1993-11-30 | Stryker Corporation | Osteogenic proteins |
US5354557A (en) | 1988-04-08 | 1994-10-11 | Stryker Corporation | Osteogenic devices |
US4968590A (en) | 1988-04-08 | 1990-11-06 | Stryker Corporation | Osteogenic proteins and polypeptides |
US5024841A (en) | 1988-06-30 | 1991-06-18 | Collagen Corporation | Collagen wound healing matrices and process for their production |
US5071834A (en) | 1988-09-16 | 1991-12-10 | Genentech, Inc. | Purified activin B composition |
US5284756A (en) | 1988-10-11 | 1994-02-08 | Lynn Grinna | Heterodimeric osteogenic factor |
US5106626A (en) | 1988-10-11 | 1992-04-21 | International Genetic Engineering, Inc. | Osteogenic factors |
US4955892A (en) | 1988-10-24 | 1990-09-11 | Louisiana State University | Neural cell adhesion protein nerve prosthesis |
US5011486A (en) | 1988-11-18 | 1991-04-30 | Brown University Research Foundation | Composite nerve guidance channels |
US5162430A (en) * | 1988-11-21 | 1992-11-10 | Collagen Corporation | Collagen-polymer conjugates |
US5510418A (en) | 1988-11-21 | 1996-04-23 | Collagen Corporation | Glycosaminoglycan-synthetic polymer conjugates |
US4920962A (en) | 1988-11-23 | 1990-05-01 | Claude Proulx | Splint-like element for use in end-to-end nerve suture |
US5217867A (en) | 1988-11-30 | 1993-06-08 | The Salk Institute For Biological Studies | Receptors: their identification, characterization, preparation and use |
US5013549A (en) | 1989-02-16 | 1991-05-07 | Board Of Trustees Operating Michigan State University | Production, isolation, and identification of novel antifungal compounds |
CA2030518C (en) | 1989-03-28 | 2000-03-21 | Elizabeth A. Wang | Osteoinductive compositions |
US5026381A (en) | 1989-04-20 | 1991-06-25 | Colla-Tec, Incorporated | Multi-layered, semi-permeable conduit for nerve regeneration comprised of type 1 collagen, its method of manufacture and a method of nerve regeneration using said conduit |
US4963146A (en) | 1989-04-20 | 1990-10-16 | Colla-Tec Incorporated | Multi-layered, semi-permeable conduit for nerve regeneration |
US5166322A (en) | 1989-04-21 | 1992-11-24 | Genetics Institute | Cysteine added variants of interleukin-3 and chemical modifications thereof |
US5399346A (en) | 1989-06-14 | 1995-03-21 | The United States Of America As Represented By The Department Of Health And Human Services | Gene therapy |
AU5958090A (en) | 1989-06-29 | 1991-01-17 | United States of America, as represented by the Secretary, U.S. Department of Commerce, The | Method for protecting bone marrow against chemotherapeutic drugs and radiation therapy using transforming growth factor beta 1 |
US5324519A (en) | 1989-07-24 | 1994-06-28 | Atrix Laboratories, Inc. | Biodegradable polymer composition |
HU204530B (en) * | 1989-08-10 | 1992-01-28 | Richter Gedeon Vegyeszet | Process for producing new 1-oxa-2-oxo-8-aza-spiro(4,5)decane derivatives and pharmaceutical compositions containing them |
US5422340A (en) | 1989-09-01 | 1995-06-06 | Ammann; Arthur J. | TGF-βformulation for inducing bone growth |
US5166190A (en) | 1990-01-08 | 1992-11-24 | Genentech, Inc. | Method for increasing fertility in males |
US5256418A (en) | 1990-04-06 | 1993-10-26 | Organogenesis, Inc. | Collagen constructs |
US5290271A (en) * | 1990-05-14 | 1994-03-01 | Jernberg Gary R | Surgical implant and method for controlled release of chemotherapeutic agents |
DE69132823T2 (en) | 1990-05-16 | 2002-07-18 | Genetics Inst | BONE AND Cartilage Formation Proton Proteins |
WO1991017777A2 (en) * | 1990-05-22 | 1991-11-28 | University Of Florida | Injectable bioactive glass compositions and methods for tissue reconstruction |
US5168050A (en) | 1990-05-24 | 1992-12-01 | Genentech, Inc. | Mammalian expression of the bone morphogenetic protein-2b using bmp2a/bmp2b fusion |
US5218090A (en) | 1990-06-12 | 1993-06-08 | Warner-Lambert Company | EGF receptor truncates |
US5364839A (en) | 1990-06-18 | 1994-11-15 | Genetics Institute, Inc. | Osteoinductive pharmaceutical formulations |
US5206028A (en) | 1991-02-11 | 1993-04-27 | Li Shu Tung | Dense collagen membrane matrices for medical uses |
US5208219A (en) | 1991-02-14 | 1993-05-04 | Celtrix Pharmaceuticals Inc. | Method for inducing bone growth |
US5118667A (en) | 1991-05-03 | 1992-06-02 | Celtrix Pharmaceuticals, Inc. | Bone growth factors and inhibitors of bone resorption for promoting bone formation |
US5229495A (en) | 1991-06-18 | 1993-07-20 | Ludwig Institute For Cancer Research | Substantially pure receptor like TGF-β 1 binding molecules and uses thereof |
US5216126A (en) | 1991-06-19 | 1993-06-01 | Genentech, Inc. | Receptor polypeptides and their production and uses |
ES2127757T3 (en) | 1991-06-25 | 1999-05-01 | Genetics Inst | COMPOSITIONS BMP-9. |
US5356629A (en) * | 1991-07-12 | 1994-10-18 | United States Surgical Corporation | Composition for effecting bone repair |
US5306307A (en) | 1991-07-22 | 1994-04-26 | Calcitek, Inc. | Spinal disk implant |
US5171579A (en) | 1991-10-11 | 1992-12-15 | Genetics Institute, Inc. | Formulations of blood clot-polymer matrix for delivery of osteogenic proteins |
JP3504263B2 (en) | 1991-11-04 | 2004-03-08 | ジェネティックス・インスチチュート・リミテッド・ライアビリティ・カンパニー | Recombinant bone morphogenetic protein heterodimers, compositions and uses |
SE469653B (en) * | 1992-01-13 | 1993-08-16 | Lucocer Ab | POROEST IMPLANT |
ATE208217T1 (en) | 1992-02-28 | 2001-11-15 | Cohesion Tech Inc | INJECTABLE CERAMIC COMPOUNDS AND METHOD FOR THEIR PRODUCTION AND USE |
IT1259090B (en) * | 1992-04-17 | 1996-03-11 | Fidia Spa | BIOMATERIALS FOR BONE PROSTHESIS |
IT1259100B (en) * | 1992-05-20 | 1996-03-11 | Lanfranco Callegaro | USE OF HYDROGELS FOR THE LOCKING OF PROSTHETIC SYSTEMS |
CZ283073B6 (en) * | 1992-11-03 | 1997-12-17 | Chemickotechnologická Fakulta Stu | Biologically active material and process for preparing thereof |
KR950008384B1 (en) * | 1992-12-10 | 1995-07-28 | 삼성전자주식회사 | Method of forming pattern |
US5525148A (en) * | 1993-09-24 | 1996-06-11 | American Dental Association Health Foundation | Self-setting calcium phosphate cements and methods for preparing and using them |
US5736160A (en) * | 1993-10-28 | 1998-04-07 | Thm Biomedical, Inc. | Process and device for treating and healing a bone void |
JP3532947B2 (en) * | 1993-11-20 | 2004-05-31 | 株式会社堀場製作所 | Apparatus for measuring concentration of exhaust gas generated from automobile and dead time correction method for gas analyzer |
JP3717930B2 (en) | 1993-12-07 | 2005-11-16 | ジェネティックス・インスチチュート・リミテッド・ライアビリティ・カンパニー | BMP-12, BMP-13 and their tendon-derived compositions |
US5399677A (en) | 1993-12-07 | 1995-03-21 | Genetics Institute, Inc. | Mutants of bone morphogenetic proteins |
DK0831884T3 (en) * | 1995-06-05 | 2003-11-03 | Inst Genetics Llc | Use of bone morphogenetic proteins for healing and repair of connective tissue binding |
US5752974A (en) * | 1995-12-18 | 1998-05-19 | Collagen Corporation | Injectable or implantable biomaterials for filling or blocking lumens and voids of the body |
BR9612051A (en) * | 1995-12-18 | 1999-02-09 | Degussa | Medical implant |
CN1149088C (en) * | 1996-03-05 | 2004-05-12 | 奥奎斯特公司 | Method of promoting bone growth with hyaluronic acid and growth factors |
NZ331238A (en) | 1996-03-05 | 2000-05-26 | Orquest Inc | Method of promoting bone growth with hyaluronic acid and growth factors (bFGF) |
WO1997045532A1 (en) * | 1996-05-28 | 1997-12-04 | Brown University Research Foundation | Hyaluronan based biodegradable scaffolds for tissue repair |
IT1288290B1 (en) * | 1996-06-21 | 1998-09-11 | Fidia Spa In Amministrazione S | SELF-LETICULATED HYALURONIC ACID AND RELATED PHARMACEUTICAL COMPOSITIONS FOR THE TREATMENT OF ARTHROPATHIES |
ATE197900T1 (en) * | 1996-08-15 | 2000-12-15 | Losan Pharma Gmbh | EASY TO SWALLOW ORAL MEDICINAL FORM |
US5813411A (en) * | 1996-08-20 | 1998-09-29 | Menlo Care, Inc. | Method of deforming tissue with a swollen hydrogel |
JP4388602B2 (en) * | 1997-02-07 | 2009-12-24 | ストライカー コーポレイション | Bone-forming device not containing matrix, graft, and method of use thereof |
IT1296689B1 (en) * | 1997-11-06 | 1999-07-14 | Fidia Advanced Biopolymers Srl | ESTERE DERIVATIVES OF HYALURONIC ACID HAVING VISCOELASTIC PROPERTIES AND THEIR USE IN THE BIOMEDICAL-HEALTH FIELD |
US6630457B1 (en) | 1998-09-18 | 2003-10-07 | Orthogene Llc | Functionalized derivatives of hyaluronic acid, formation of hydrogels in situ using same, and methods for making and using same |
IT1302534B1 (en) * | 1998-12-21 | 2000-09-05 | Fidia Advanced Biopolymers Srl | INJECTABLE, BIOCOMPATIBLE AND BIODEGRADABLE COMPOSITIONS INCLUDING AT LEAST A DERIVATIVE OF HYALURONIC ACID, CHONDROGENIC CELLS, FOR |
JP4211108B2 (en) | 1999-01-13 | 2009-01-21 | 生化学工業株式会社 | High viscoelastic injection device |
PT1223990E (en) | 1999-10-15 | 2004-12-31 | Fidia Advanced Biopolymers Srl | HYALURONIC ACID FORMULATIONS FOR ADMINISTRATION OF OSTEOGENIC PROTEINS |
US6599516B1 (en) * | 2000-09-14 | 2003-07-29 | Etex Corporation | Malleable implant containing solid element that resorbs or fractures to provide access channels |
US6547866B1 (en) * | 2000-10-30 | 2003-04-15 | Howmedica Osteonics Corp. | Porous calcium phosphate cement |
EP1370287A2 (en) | 2000-12-01 | 2003-12-17 | Wyeth | Method and composition for modulating bone growth |
EP1519744A4 (en) * | 2002-05-17 | 2007-10-03 | Wyeth Corp | Injectable solid hyaluronic acid carriers for delivery of osteogenic proteins |
-
2000
- 2000-10-13 PT PT00970914T patent/PT1223990E/en unknown
- 2000-10-13 DK DK00970914T patent/DK1223990T3/en active
- 2000-10-13 JP JP2001531430A patent/JP4703926B2/en not_active Expired - Fee Related
- 2000-10-13 AU AU80230/00A patent/AU774427B2/en not_active Ceased
- 2000-10-13 WO PCT/US2000/028468 patent/WO2001028602A1/en active Search and Examination
- 2000-10-13 AT AT00970914T patent/ATE271886T1/en not_active IP Right Cessation
- 2000-10-13 CA CA002386408A patent/CA2386408A1/en not_active Abandoned
- 2000-10-13 US US09/687,281 patent/US7189392B1/en not_active Expired - Fee Related
- 2000-10-13 EP EP00970914A patent/EP1223990B1/en not_active Revoked
- 2000-10-13 DE DE60012557T patent/DE60012557T2/en not_active Revoked
- 2000-10-13 ES ES00970914T patent/ES2225241T3/en not_active Expired - Lifetime
- 2000-10-13 EP EP10182996A patent/EP2286847A1/en not_active Withdrawn
-
2004
- 2004-07-30 AU AU2004203514A patent/AU2004203514B2/en not_active Ceased
-
2007
- 2007-01-18 US US11/624,463 patent/US7608580B2/en not_active Expired - Fee Related
Also Published As
Publication number | Publication date |
---|---|
ATE271886T1 (en) | 2004-08-15 |
JP4703926B2 (en) | 2011-06-15 |
ES2225241T3 (en) | 2005-03-16 |
US7608580B2 (en) | 2009-10-27 |
EP2286847A1 (en) | 2011-02-23 |
PT1223990E (en) | 2004-12-31 |
AU774427B2 (en) | 2004-06-24 |
US20070134342A1 (en) | 2007-06-14 |
AU8023000A (en) | 2001-04-30 |
AU2004203514A1 (en) | 2004-08-26 |
EP1223990A1 (en) | 2002-07-24 |
JP2003512341A (en) | 2003-04-02 |
WO2001028602A1 (en) | 2001-04-26 |
DE60012557T2 (en) | 2005-08-04 |
AU2004203514B2 (en) | 2006-08-10 |
EP1223990B1 (en) | 2004-07-28 |
DE60012557D1 (en) | 2004-09-02 |
US7189392B1 (en) | 2007-03-13 |
DK1223990T3 (en) | 2004-11-29 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
AU774427B2 (en) | Formulations of hyaluronic acid for delivery of osteogenic proteins | |
US5385887A (en) | Formulations for delivery of osteogenic proteins | |
JP3336010B2 (en) | Formulation of clot-polymer matrix for delivery of osteogenic proteins | |
US7875590B2 (en) | Injectable solid hyaluronic acid carriers for delivery of osteogenic proteins | |
JP2004262758A (en) | Porous beta-tricalcium phosphate granule and method for producing the same | |
EP1220693B1 (en) | Formulations for delivery of osteogenic proteins | |
WO1994015653A1 (en) | Tgf-beta formulation for inducing bone growth | |
EP1454640A2 (en) | Formulations of hyaluronic acid for delivery of osteogenic proteins | |
AU8023000B2 (en) | ||
EP1475109A1 (en) | Formulations for delivery of osteogenic proteins | |
KR20050019075A (en) | Injectable solid hyaluronic acid carriers for delivery of osteogenic proteins |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
EEER | Examination request | ||
FZDE | Discontinued |
Effective date: 20130528 |
|
FZDE | Discontinued |
Effective date: 20130528 |