Method for promoting bone growth using activin-actriia antagonists
In certain aspects, the present invention provides compositions and methods for promoting bone growth and increasing bone density.
1. A method for treating bone fractures, comprising administering to a subject in need thereof an effective amount of a polypeptide comprising an amino acid sequence that is at least 95% identical to SEQ ID NO: 2.
2. The method of claim 1 , further comprising administering a second bone active-agent.
3. The method of claim 2 , wherein the bone active-agent is selected from a bisphosphonate, an estrogen, a selective estrogen receptor modulator, a parathyroid hormone, a calcitonin, a calcium supplement, and a vitamin D supplement.
4. The method of claim 2 , wherein the second bone active-agent is administered conjointly with the polypeptide.
5. The method of claims 4 , wherein the polypeptide and the second bone active-agent are administered as a single co-formulation.
6. The method of claim 4 , wherein the polypeptide and the second bone active-agent are administered simultaneously.
7. The method of claim 4 , wherein the polypeptide and the second bone active-agent are administered at separate times.
8. The method of claim 1 , wherein the polypeptide includes one or more modified amino acid residues selected from a glycosylated amino acid, a PEGylated amino acid, a farnesylated amino acid, an acetylated amino acid, a biotinylated amino acid, or an amino acid conjugated to a lipid moiety.
9. The method of claim 1 , wherein the polypeptide is administered no more frequently than once per month.
10. The method of claim 1 , wherein the polypeptide is administered no more frequently than once every three months.
11. The method of claim 1 , wherein the polypeptide is administered no more frequently than once every six months.
12. The method of claim 1 , wherein the polypeptide is at least 98% pure as determined by size-exclusion chromatography.
13. The method of claim 1 , wherein the polypeptide comprises an amino acid sequence that is at least 97% identical to SEQ ID NO: 2.
14. The method of claim 1 , wherein the polypeptide comprises an amino acid sequence that is at least 99% identical to SEQ ID NO: 2.
15. The method of claim 1 , wherein the polypeptide comprises the amino acid sequence of SEQ ID NO: 2.
16. The method of claim 1 , wherein the polypeptide is glycosylated and has a glycosylation pattern identical to a glycosylation pattern obtained by expressing the polypeptide in a Chinese hamster ovary (CHO) cell line.
17. The method of claim 1 , wherein the N-terminus of the polypeptide is ILGRSETQE (SEQ ID NO: 11).
18. The method of claim 1 , wherein the polypeptide further comprises a domain that enhances one or more of in vivo stability, in vivo half-life, uptake/administration, tissue localization or distribution, or formation of protein complexes.
19. The method of claim 18 , wherein the domain is an immunoglobulin Fc domain.
20. The method of claim 18 , wherein the domain is serum albumin.
21. The method of claim 1 , wherein the serum half-life of the polypeptide is greater than two weeks.
22. The method of claim 1 , wherein the serum half-life of the polypeptide is 20-30 days.
23. The method of claim 1 , wherein the polypeptide has one or more of the following characteristics: (i) binds to an ActRIIa ligand with a K D of at least 10 −7 M; and (ii) inhibits ActRIIa signaling.
24. The method of claim 1 , wherein the polypeptide is soluble.
25. The method of claim 1 , wherein the polypeptide is an antagonist of activin A.
26. The method of claim 1 , wherein the polypeptide comprises an amino acid sequence that is at least 95% identical to SEQ ID NO: 7.
27. The method of claim 1 , wherein the polypeptide comprises an amino acid sequence that is at least 97% identical to SEQ ID NO: 7.
28. The method of claim 1 , wherein the polypeptide comprises an amino acid sequence that is at least 99% identical to SEQ ID NO: 7.
29. The method of claim 1 , wherein the polypeptide comprises the amino acid sequence of SEQ ID NO: 7.
30. The method of claim 1 , wherein the polypeptide binds to activin.
31. The method of claim 1 , wherein the polypeptide is recombinantly expressed using a Tissue Plasminogen Activator (TPA) leader sequence.
32. The method of claim 31 , wherein the TPA leader sequence comprises the amino acid sequence of SEQ ID NO:9.
33. A method for increasing bone strength, comprising administering to a subject in need thereof an effective amount of a polypeptide comprising an amino acid sequence that is at least 95% identical to SEQ ID NO: 2.
34. The method of claim 33 , further comprising administering a second bone active-agent.
35. The method of claim 34 , wherein the bone active-agent is selected from a bisphosphonate, an estrogen, a selective estrogen receptor modulator, a parathyroid hormone, a calcitonin, a calcium supplement, and a vitamin D supplement.
36. The method of claim 34 , wherein the second bone active-agent is administered conjointly with the polypeptide.
37. The method of claims 34 , wherein the polypeptide and the second bone active-agent are administered as a single co-formulation.
38. The method of claim 34 , wherein the polypeptide and the second bone active-agent are administered simultaneously.
39. The method of claim 34 , wherein the polypeptide and the second bone active-agent are administered at separate times.
40. The method of claim 33 , wherein the polypeptide includes one or more modified amino acid residues selected from a glycosylated amino acid, a PEGylated amino acid, a farnesylated amino acid, an acetylated amino acid, a biotinylated amino acid, or an amino acid conjugated to a lipid moiety.
41. The method of claim 33 , wherein the polypeptide is administered no more frequently than once per month.
42. The method of claim 33 , wherein the polypeptide is administered no more frequently than once every three months.
43. The method of claim 33 , wherein the polypeptide is administered no more frequently than once every six months.
44. The method of claim 33 , wherein the polypeptide is at least 98% pure as determined by size-exclusion chromatography.
45. The method of claim 33 , wherein the polypeptide comprises an amino acid sequence that is at least 97% identical to SEQ ID NO: 2.
46. The method of claim 33 , wherein the polypeptide comprises an amino acid sequence that is at least 99% identical to SEQ ID NO: 2.
47. The method of claim 33 , wherein the polypeptide comprises the amino acid sequence of SEQ ID NO: 2.
48. The method of claim 33 , wherein the polypeptide is glycosylated and has a glycosylation pattern identical to a glycosylation pattern obtained by expressing the polypeptide in a Chinese hamster ovary (CHO) cell line.
49. The method of claim 33 , wherein the N-terminus of the polypeptide is ILGRSETQE (SEQ ID NO: 11).
50. The method of claim 33 , wherein the polypeptide further comprises a domain that enhances one or more of in vivo stability, in vivo half-life, uptake/administration, tissue localization or distribution, or formation of protein complexes.
51. The method of claim 50 , wherein the domain is an immunoglobulin Fc domain.
52. The method of claim 50 , wherein the domain is serum albumin.
53. The method of claim 33 , wherein the serum half-life of the polypeptide is greater than two weeks.
54. The method of claim 33 , wherein the serum half-life of the polypeptide is 20-30 days.
55. The method of claim 33 , wherein the polypeptide has one or more of the following characteristics: (i) binds to an ActRIIa ligand with a K D of at least 10 −7 M; and (ii) inhibits ActRIIa signaling.
56. The method of claim 33 , wherein the polypeptide is soluble.
57. The method of claim 33 , wherein the polypeptide binds to activin.
58. The method of claim 33 , wherein the polypeptide is an antagonist of activin A.
59. The method of claim 33 , wherein the polypeptide comprises an amino acid sequence that is at least 95% identical to SEQ ID NO: 7.
60. The method of claim 33 , wherein the polypeptide comprises an amino acid sequence that is at least 97% identical to SEQ ID NO: 7.
61. The method of claim 33 , wherein the polypeptide comprises an amino acid sequence that is at least 99% identical to SEQ ID NO: 7.
62. The method of claim 33 , wherein the polypeptide comprises the amino acid sequence of SEQ ID NO: 7.
63. The method of claim 33 , wherein the polypeptide is recombinantly expressed using a Tissue Plasminogen Activator (TPA) leader sequence.
64. The method of claim 63 , wherein the TPA leader sequence comprises the amino acid sequence of SEQ ID NO:9.
65. A method for increasing bone mineral density comprising administering to a subject in need thereof an effective amount of a polypeptide comprising an amino acid sequence that is at least 95% identical to SEQ ID NO: 2.
66. The method of claim 65 , further comprising administering a second bone active-agent.
67. The method of claim 66 , wherein the bone-active agent is selected from a bisphosphonate, an estrogen, a selective estrogen receptor modulator, a parathyroid hormone, a calcitonin, a calcium supplement, and a vitamin D supplement.
68. The method of claim 66 , wherein the second bone active-agent is administered conjointly with the polypeptide.
69. The method of claims 66 , wherein the polypeptide and the second bone active-agent are administered as a single co-formulation.
70. The method of claim 66 , wherein the polypeptide and the second bone active-agent are administered simultaneously.
71. The method of claim 66 , wherein the polypeptide and the second bone active-agent are administered at separate times.
72. The method of claim 65 , wherein the polypeptide includes one or more modified amino acid residues selected from a glycosylated amino acid, a PEGylated amino acid, a farnesylated amino acid, an acetylated amino acid, a biotinylated amino acid, or an amino acid conjugated to a lipid moiety.
73. The method of claim 65 , wherein the polypeptide is administered no more frequently than once per month.
74. The method of claim 65 , wherein the polypeptide is administered no more frequently than once every three months.
75. The method of claim 65 , wherein the polypeptide is administered no more frequently than once every six months.
76. The method of claim 65 , wherein the polypeptide is at least 98% pure as determined by size-exclusion chromatography.
77. The method of claim 65 , wherein the polypeptide comprises an amino acid sequence that is at least 97% identical to SEQ ID NO: 2.
78. The method of claim 65 , wherein the polypeptide comprises an amino acid sequence that is at least 99% identical to SEQ ID NO: 2.
79. The method of claim 65 , wherein the polypeptide comprises the amino acid sequence of SEQ ID NO: 2.
80. The method of claim 65 , wherein the polypeptide is glycosylated and has a glycosylation pattern identical to a glycosylation pattern obtained by expressing the polypeptide in a Chinese hamster ovary (CHO) cell line.
81. The method of claim 65 , wherein the N-terminus of the polypeptide is ILGRSETQE (SEQ ID NO: 11).
82. The method of claim 65 , wherein the polypeptide further comprises a domain that enhances one or more of in vivo stability, in vivo half-life, uptake/administration, tissue localization or distribution, or formation of protein complexes.
83. The method of claim 82 , wherein the domain is an immunoglobulin Fc domain.
84. The method of claim 82 , wherein the domain is serum albumin.
85. The method of claim 65 , wherein the serum half-life of the polypeptide is greater than two weeks.
86. The method of claim 65 , wherein the serum half-life of the polypeptide is 20-30 days.
87. The method of claim 65 , wherein the polypeptide has one or more of the following characteristics: (i) binds to an ActRIIa ligand with a K D of at least 10 −7 M; and (ii) inhibits ActRIIa signaling.
88. The method of claim 65 , wherein the polypeptide is soluble.
89. The method of claim 65 , wherein the polypeptide binds to activin.
90. The method of claim 65 , wherein the polypeptide is an antagonist of activin A.
91. The method of claim 65 , wherein the polypeptide comprises an amino acid sequence that is at least 95% identical to SEQ ID NO: 7.
92. The method of claim 65 , wherein the polypeptide comprises an amino acid sequence that is at least 97% identical to SEQ ID NO: 7.
93. The method of claim 65 , wherein the polypeptide comprises an amino acid sequence that is at least 99% identical to SEQ ID NO: 7.
94. The method of claim 65 , wherein the polypeptide comprises the amino acid sequence of SEQ ID NO: 7.
95. The method of claim 65 , wherein the polypeptide is recombinantly expressed using a Tissue Plasminogen Activator (TPA) leader sequence.
96. The method of claim 95 , wherein the TPA leader sequence comprises the amino acid sequence of SEQ ID NO:9.
97. A method for treating a bone-related disorder, comprising administering to a subject in need thereof an effective amount of a polypeptide comprising an amino acid sequence that is at least 95% identical to SEQ ID NO: 2, wherein the bone-related disorder is associated with low bone density or decreased bone strength.
98. The method of claim 97 , wherein the bone-related disorder is selected from osteoporosis, hypogonadal bone loss, tumor-induced bone loss, cancer therapy-induced bone loss, bony metastases, hyperparathyroidism, Cushing's disease, thyrotoxicosis, chronic diarrheal state or malabsorption, renal tubular acidosis, anorexia nervosa, and Paget's disease.
99. The method of claim 97 , further comprising administering a second bone active-agent.
100. The method of claim 99 , wherein the bone active-agent is selected from a bisphosphonate, an estrogen, a selective estrogen receptor modulator, a parathyroid hormone, a calcitonin, a calcium supplement, and a vitamin D supplement.
101. The method of claim 99 , wherein the second bone active-agent is administered conjointly with the polypeptide.
102. The method of claims 99 , wherein the polypeptide and the second bone active-agent are administered as a single co-formulation.
103. The method of claim 99 , wherein the polypeptide and the second bone active-agent are administered simultaneously.
104. The method of claim 99 , wherein the polypeptide and the second bone active-agent are administered at separate times.
105. The method of claim 97 , wherein the polypeptide includes one or more modified amino acid residues selected from a glycosylated amino acid, a PEGylated amino acid, a farnesylated amino acid, an acetylated amino acid, a biotinylated amino acid, or an amino acid conjugated to a lipid moiety.
106. The method of claim 97 , wherein the polypeptide is administered no more frequently than once per month.
107. The method of claim 97 , wherein the polypeptide is administered no more frequently than once every three months.
108. The method of claim 97 , wherein the polypeptide is administered no more frequently than once every six months.
109. The method of claim 97 , wherein the polypeptide is at least 98% pure as determined by size-exclusion chromatography.
110. The method of claim 97 , wherein the polypeptide comprises an amino acid sequence that is at least 97% identical to SEQ ID NO: 2.
111. The method of claim 97 , wherein the polypeptide comprises an amino acid sequence that is at least 99% identical to SEQ ID NO: 2.
112. The method of claim 97 , wherein the polypeptide comprises the amino acid sequence of SEQ ID NO: 2.
113. The method of claim 97 , wherein the polypeptide is glycosylated and has a glycosylation pattern identical to a glycosylation pattern obtained by expressing the polypeptide in a Chinese hamster ovary (CHO) cell line.
114. The method of claim 97 , wherein the N-terminus of the polypeptide is ILGRSETQE (SEQ ID NO: 11).
115. The method of claim 97 , wherein the polypeptide further comprises a domain that enhances one or more of in vivo stability, in vivo half-life, uptake/administration, tissue localization or distribution, or formation of protein complexes.
116. The method of claim 115 , wherein the domain is an immunoglobulin Fc domain.
117. The method of claim 115 , wherein the domain is serum albumin.
118. The method of claim 97 , wherein the serum half-life of the polypeptide is greater than two weeks.
119. The method of claim 97 , wherein the serum half-life of the polypeptide is 20-30 days.
120. The method of claim 97 , wherein the polypeptide has one or more of the following characteristics: (i) binds to an ActRIIa ligand with a K D of at least 10 −7 M; and (ii) inhibits ActRIIa signaling.
121. The method of claim 97 , wherein the polypeptide is soluble.
122. The method of claim 97 , wherein the polypeptide binds to activin.
123. The method of claim 97 , wherein the polypeptide is an antagonist of activin A.
124. The method of claim 97 , wherein the polypeptide comprises an amino acid sequence that is at least 95% identical to SEQ ID NO: 7.
125. The method of claim 97 , wherein the polypeptide comprises an amino acid sequence that is at least 97% identical to SEQ ID NO: 7.
126. The method of claim 97 , wherein the polypeptide comprises an amino acid sequence that is at least 99% identical to SEQ ID NO: 7.
127. The method of claim 97 , wherein the polypeptide comprises the amino acid sequence of SEQ ID NO: 7.
128. The method of claim 97 , wherein the polypeptide is recombinantly expressed using a Tissue Plasminogen Activator (TPA) leader sequence.
129. The method of claim 128 , wherein the TPA leader sequence comprises the amino acid sequence of SEQ ID NO:9.