FUSION PROTEINS OF COLLAGEN-BINDING DOMAIN AND PARATHYROID HORMONE

• US 20100129341A1
• Filed: 04/09/2008
• Published: 05/27/2010
• Est. Priority Date: 04/09/2007
• Status: Active Grant

• Current Assignee: Ochsner Clinic Foundation (Humana Incorporated), National University Corporation Kagawa University, The Board of Trustees of the University of Arkansas (University of Arkansas System)
• Original Assignee: The Board of Trustees of the University of Arkansas (University of Arkansas System)
• Inventors: Sakon, Joshua, Gensure, Robert C., Matsushita, Osamu
• Granted Patent: US 8,450,273 B2
• Time in Patent Office: Days
• US Class Current: 424/94.600
• CPC Class Codes: A61K 2800/86 Products or compounds obtained by genetic engineering
• CPC Class Codes: A61K 2800/91 Injection
• CPC Class Codes: A61K 35/28 Bone marrow; Haematopoietic stem cells; Mesenchymal stem cells of any origin, e.g. adipose-derived stem cells
• CPC Class Codes: A61K 38/164 from bacteria
• CPC Class Codes: A61K 38/29 Parathyroid hormone, i.e. parathormone; Parathyroid hormone-related peptides
• CPC Class Codes: A61K 38/4886 Metalloendopeptidases (3.4.24), e.g. collagenase
• CPC Class Codes: A61K 8/64 Proteins; Peptides; Derivatives or degradation products thereof
• CPC Class Codes: A61K 8/66 Enzymes
• CPC Class Codes: A61K 9/0019 Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner non-active ingredients are additionally classified in A61K47/00
• CPC Class Codes: A61P 13/12 of the kidneys
• CPC Class Codes: A61P 17/14 for baldness or alopecia
• CPC Class Codes: A61P 19/00 Drugs for skeletal disorders
• CPC Class Codes: A61P 19/08 for bone diseases, e.g. rachitism, Paget's disease
• CPC Class Codes: A61P 19/10 for osteoporosis
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 35/04 specific for metastasis
• CPC Class Codes: A61P 37/04 Immunostimulants
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: A61P 5/18 of the parathyroid hormones
• CPC Class Codes: A61P 5/20 for decreasing, blocking or antagonising the activity of PTH
• CPC Class Codes: A61P 9/10 : for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
• CPC Class Codes: A61Q 7/00 : Preparations for affecting hair growth
• CPC Class Codes: C07K 14/635 : Parathyroid hormone, i.e. parathormone; Parathyroid hormone-related peptides
• CPC Class Codes: C07K 2319/00 : Fusion polypeptide
• CPC Class Codes: C07K 2319/50 : containing protease site
• CPC Class Codes: C07K 2319/70 : containing domain for protein-protein interaction
• CPC Class Codes: C12N 9/1088 : Glutathione transferase (2.5.1.18)
• CPC Class Codes: C12N 9/52 : derived from bacteria or Archaea
• CPC Class Codes: C12N 9/64 : derived from animal tissue
• CPC Class Codes: C12N 9/6489 : Metalloendopeptidases (3.4.24)
• CPC Class Codes: C12Y 205/01018 : Glutathione transferase (2.5.1.18)
• CPC Class Codes: C12Y 304/24003 : Microbial collagenase (3.4.24.3)
• CPC Class Codes: C12Y 304/24007 : Interstitial collagenase (3.4.24.7), i.e. matrix metalloprotease 1 or MMP1

Abstract

Fusion proteins containing active agonist or antagonist fragments of parathyroid hormone (PTH) and parathyroid hormone related peptide (PTHrP) coupled to a collagen-binding domain are presented. The fusion proteins can be used to promote bone growth, to promote hair growth, to prevent cancer metastasis to bone, to promote immune reconstitution with a bone marrow stem cell transplant, to promote mobilization of bone marrow stem cells for collection for autologous stem cell transplant, and to treat renal osteodystrophy. Pharmaceutical agents comprising a collagen-binding polypeptide segment linked to a non-peptidyl PTH/PTHrP receptor agonist or antagonist are also presented.

Claims

1-60. -60. (canceled)

61. A composition comprising:

a collagen-binding polypeptide segment linked to

a PTH/PTHrP receptor agonist;

wherein the collagen-binding polypeptide segment is a bacterial collagen binding polypeptide segment or a segment of a collagenase.

62. The composition of claim 61, wherein the collagen-binding polypeptide segment and the PTH/PTHrP receptor agonist are chemically cross-linked to each other or are polypeptide portions of a fusion protein.

63. The composition of claim 61,

wherein the composition has at least 50% greater activity than PTH(1-34) as measured by the method of Example 4.

64. The composition of claim 61, wherein the collagen-binding polypeptide segment comprises residues 38-158 of SEQ ID NO:

I, a fragment of at least 8 consecutive amino acids from residues 38-158 of SEQ ID NO;

I, or is at least 90% identical to residues 38-158 of SEQ ID NO;

1.

65. The composition of claim 61, wherein the PTH/PTHrP receptor agonist comprises residues 1-33 of SEQ ID NO:

I, PTH (SEQ ID NO;

     7), residues 1-14 of SEQ ID NO;

1, residues 1-34 of SEQ ID NO;

7 or a fragment of at least 8 consecutive amino acids from residues 1-34 of SEQ ID NO;

7.

66. The composition of claim 61, wherein the PTH/PTHrP receptor agonist is a polypeptide and the N-terminus of the collagen-binding polypeptide segment is linked directly or through a linker polypeptide segment to the C-terminus of the PTH/PTHrP receptor agonist polypeptide.

67. A composition comprising:

a collagen-binding polypeptide segment linked to a PTH/PTHrP receptor agonist;

wherein the composition has at least 50% greater activity than PTH(1-34) as measured by the method of Example 4.

68. The composition of claim 67, wherein the collagen-binding polypeptide segment and the PTH/PTHrP receptor agonist are chemically cross-linked to each other or are polypeptide portions of a fusion protein.

69. The composition of claim 67, wherein the collagen-binding polypeptide segment comprises residues 38-158 of SEQ ID NO:

I, a fragment of at least 8 consecutive amino acids from residues 38-158 of SEQ ID NO;

I, or is at least 90% identical to residues 38-158 of SEQ ID NO;

1.

70. The composition of claim 67, wherein the PTH/PTHrP receptor agonist comprises residues 1-33 of SEQ ID NO:

I, PTH (SEQ ID NO;

     7), residues 1-14 of SEQ ID NO;

1, residues 1-34 of SEQ ID NO;

7 or a fragment of at least 8 consecutive amino acids from residues 1-34 of SEQ ID NO;

7.

71. The composition of claim 67, wherein the PTH/PTHrP receptor agonist is a polypeptide and the N-terminus of the collagen-binding polypeptide segment is linked directly or through a linker polypeptide segment to the C-terminus of the PTH/PTHrP agonist polypeptide segment.

72. A composition comprising:

a collagen-binding polypeptide segment linked to

a PTH/PTHrP receptor antagonist.

73. The composition of claim 72, wherein the collagen-binding polypeptide segment and the PTH/PTHrP receptor antagonist are chemically cross-linked to each other or are polypeptide portions of a fusion protein.

74. The composition of claim 72, wherein the collagen-binding polypeptide segment comprises residues 38-158 of SEQ ID NO:

I, a fragment of at least 8 consecutive amino acids from residues 38-158 of SEQ ID NO;

I, or is at least 90% identical to residues 38-158 of SEQ ID NO;

1.

75. The composition of claim 72, wherein the PTH/PTHrP receptor antagonist comprises residues 7-34 of SEQ ID NO:

7, is a fragment of at least 8 consecutive amino acids from residues 7-34 of SEQ ID NO;

7, or comprises residues 7-14 of SEQ ID NO;

7.

83. A method of attaining an outcome comprising administering to a mammal the composition of claim 72, wherein the outcome is selected from the group consisting of treating or preventing renal osteodystrophy, preventing or reducing incidence of bone metastasis in cancer, slowing the growth of metastatic cancer in bone and promoting hair growth.

76. A method of attaining an outcome comprising administering to a mammal an effective amount of a composition comprising a collagen-binding polypeptide segment linked to a PTH/PTHrP receptor agonist, wherein the outcome is selected from the group consisting of promoting bone growth, promoting hair growth, promoting tissue growth, promoting immune reconstitution, promoting bone marrow stem cell mobilization and treating myocardial infarction.

77. The method of claim 76, wherein the collagen-binding polypeptide segment is a bacterial collagen binding polypeptide segment or a segment of a collagenase.

78. The method of claim 76, wherein administering the composition to the mammal increases trabecular bone mineral density or cortical bone mineral density or trabecular bone mineral volume or cortical bone mineral volume.

79. The method of claim 78, wherein the composition has at least 50% greater activity than PTH(1-34) as measured by the method of Example 4.

80. The method of claim 76, wherein the composition is administered before, during or after administration of an implant or in combination with the implant.

81. The method of claim 76, wherein the composition is administered by injection.

82. The method of claim 76, wherein the composition is administered in aqueous solution at pH below about 5.0.