Compositions and Methods for Increasing Bone Mineralization
A novel class or family of TGF-β binding proteins is disclosed. Also disclosed are assays for selecting molecules for increasing bone mineralization and methods for utilizing such molecules.
1 - 87 . (canceled)
88 . A method of increasing bone mineral content in a warm-blooded animal in need thereof, comprising administering to said animal a therapeutically effective amount of a molecule that inhibits the activity of any one of SEQ ID NOS: 2, 6, 8, 10, 12, 14 or 16.
89 . The method of claim 88 wherein the molecule is selected from the group consisting of antibodies, antibody fragments, nucleic acids, polypeptides and peptides.
90 . The method of claim 88 wherein the molecule is selected from the group consisting of:
(a) polypeptides that bind to an amino acid sequence selected from the group consisting of SEQ ID NOs: 2, 6, 8, 10, 12, 14 and 16 with a K a of greater than or equal to 10 6 M −1 ;
(b) polypeptides that bind with a K a of greater than or equal to 10 6 M −1 to an amino acid sequence encoded by a naturally occurring polynucleotide that (i) encodes a protein that decreases bone mineral content and (ii) hybridizes under stringent conditions to any one of SEQ ID NOS: 1, 5, 7, 9, 11, 13 and 15 or the complements thereof, where the stringent conditions comprise hybridization in 5×SSPE, 5× Denhardt's solution and 0.5% SDS overnight at 55 to 60° C.;
(c) oligonucleotides that comprises a polynucleotide sequence that hybridizes under stringent conditions to any one of SEQ ID NOS: 1, 5, 7, 9, 11, 13 and 15 or the complements thereof, wherein the stringent conditions comprise hybridization in 5×SSPE, 5× Denhardt's solution and 0.5% SDS overnight at 55 to 60° C.;
(d) antibodies that bind to an amino acid sequence selected from the group consisting of SEQ ID NOs: 2, 6, 8, 10, 12, 14 and 16 with a K a of greater than or equal to 10 6 M −1 ;
(e) antibodies that bind with a K a of greater than or equal to 10 6 M −1 to an amino acid sequence encoded by a naturally occurring polynucleotide that (i) encodes a protein that decreases bone mineral content and (ii) hybridizes under stringent conditions to any one of SEQ ID NOS: 1, 5, 7, 9, 11, 13 and 15 or the complements thereof, wherein the stringent conditions comprise hybridization in 5×SSPE, 5× Denhardt's solution and 0.5% SDS overnight at 55 to 60° C.; and
(f) polypeptides comprising an antibody fragment of any of the antibodies of (d) or (e).
91 . A method of treating a human with a condition which results in loss of bone mineral content comprising administering to said human a therapeutically effective amount of an organic molecule that inhibits the activity of SEQ ID NO: 2.
92 . The method according to claim 91 , wherein the human has a dysplasia associated with abnormal growth or development of bone.
93 . The method according to claim 91 , wherein the condition is selected from the group consisting of achondroplasia, cleidocranial dysostosis, enchondromatosis, fibrous dysplasia, Gaucher's, hypophosphatemic rickets, Marfan's, multiple hereditary exotoses, neurofibromatosis, osteogenesis imperfecta, osteopetrosis, osteopoikilosis, sclerotic lesions, fractures, periodontal disease, pseudoarthrosis and pyogenic osteomyelitis.
94 . A method of increasing bone mineral content in a human in need thereof, comprising administering to said human a therapeutically effective amount of an organic molecule that inhibits the activity of SEQ ID NO: 2.
95 . The method of claim 94 wherein the human has a condition selected from the group consisting of osteopenia, osteoporosis, bone fracture, osteomalacia, periodontal disease, scurvy, Cushing's disease, conditions due to limb immobilization and conditions due to steroid usage.
96 . The method of claim 95 wherein the osteopenia is caused by an anemic state, steroids, heparin, a bone marrow disorder, scurvy, malnutrition, calcium deficiency, idiopathic osteoporosis, congenital osteopenia or osteoporosis, alcoholism, chronic liver disease, senility, post menstrual state, oligomenorrhea, amenorrhea, pregnancy, diabetes mellitus, hyperthyroidism, Cushing's disease, acromegaly, hypogonadism, immobilization or disuse, reflex sympathetic dystrophy syndrome, transient regional osteoporosis or osteomalacia.
97 . The method of claim 91 wherein the molecule is selected from the group consisting of antibodies, antibody fragments, nucleic acids, polypeptides and peptides.
98 . The method of claim 94 wherein the molecule is selected from the group consisting of antibodies, antibody fragments, nucleic acids, polypeptides and peptides.
99 . The method of any one of claim 91 or 94 , wherein the molecule is a polypeptide that binds with a K a of greater than or equal to 10 6 M −1 to an amino acid sequence encoded by a naturally occurring polynucleotide that (i) encodes a protein that decreases bone mineral content and (ii) hybridizes under stringent conditions to SEQ ID NO:
1 or the complement thereof, wherein the stringent conditions comprise hybridization in 5×SSPE, 5× Denhardt's solution and 0.5% SDS overnight at 55 to 60° C.
100 . The method of any one of claim 91 or 94 wherein the molecule is a polypeptide comprising (a) an antibody or (b) an antibody fragment, wherein said polypeptide binds to SEQ ID NO: 2 with a K a of greater than or equal to 10 6 M −1 .
101 . The method of claim 97 , wherein the antibody fragment is selected from the group consisting of F(ab′) 2 , F(ab) 2 , Fab′, Fab, and Fv.
102 . The method of claim 97 , wherein the antibody is selected from the group consisting of murine monoclonal antibodies, human monoclonal antibodies, humanized monoclonal antibodies, and antibody fragments thereof.
103 . The method of any one of claim 91 or 94 , wherein the molecule is an oligonucleotide that (i) decreases expression of SEQ ID NO: 1 and (ii) comprises a polynucleotide sequence that hybridizes under stringent conditions to SEQ ID NO: 1 or the complement thereof, wherein the stringent conditions comprise hybridization in 5×SSPE, 5× Denhardt's solution and 0.5% SDS overnight at 55 to 60° C.
104 . The method of claim 97 , wherein the nucleic acid is a ribozyme or an antisense oligonucleotide.
105 . The method of claim 97 , further comprising the step of administering another inhibitor of bone resorption.
106 . The method of claim 105 , wherein the other inhibitor of bone resorption is selected from the group consisting of calcitonin, estrogen, bisphosphonates, growth factors having anti-resorptive activity and tamoxifen.