Osteoinductive factors

• US 4,877,864 A
• Filed: 03/26/1987
• Issued: 10/31/1989
• Est. Priority Date: 03/26/1987
• Status: Expired due to Term

• Current Assignee: Genetics Institute LLC (Pfizer Inc.)
• Original Assignee: Genetics Institute LLC (Pfizer Inc.)
• Inventors: Rosen, Vicki, Wozney, John M., Wang, Elizabeth A.
• Primary Examiner(s): Tanenholtz, Alvin E.
• Application Number:
  US07/031,346
• Time in Patent Office: 950 Days
• Field of Search: 530/324, 514/12, 435/172.3, 435/70
• US Class Current: 514/17.1
• CPC Class Codes: A61K 35/32 Bones; Osteocytes; Osteoblasts; Tendons; Tenocytes; Teeth; Odontoblasts; Cartilage; Chondrocytes; Synovial membrane
• CPC Class Codes: A61K 38/00 Medicinal preparations containing peptides peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00
• CPC Class Codes: A61L 24/043 Mixtures of macromolecular materials
• CPC Class Codes: A61L 27/227 Other specific proteins or polypeptides not covered by A61L27/222, A61L27/225 or A61L27/24
• CPC Class Codes: C07K 14/51 Bone morphogenetic factor; Osteogenins; Osteogenic factor; Bone-inducing factor
• CPC Class Codes: C08L 89/00 Compositions of proteins; Compositions of derivatives thereof foodstuff preparations A23J3/00

Abstract

Human and bovine bone inductive factors are provided. Such factors may be produced by recombinant techniques and be useful in the treatment of bone defects.

Claims

1. A purified cartilage and bone inductive protein produced by the steps comprising:

(a) culturing a cell transformed with a DNA sequence comprising the sequences as follows;

GCC TTT CTT GGG GAC ATT GCC CTG GAC GAA GAG GAC CTG AGG GCC TTC CAG GTA CAG CAG GCT GTG TAT CTC AGA CGG CAC ACA GCT CGT AAG TCC TCC ATC AAA GCT GCA, naturally occuring allelic sequences thereof, equivalent degenerative codon sequences thereof or sequences which

(1) hybridize thereto under stringent hybridization condition; and

(2) upon expression encode a protein characterized by the ability to induce bone and/or cartilage formation in the in vivo rat bone formation assay at a concentration of between about 10-1000 nanograms per gram bone; and

(b) recovering and purifying from said culture medium said protein so as to be substantially free from association with other proteinaceous materials and characterized by a peptide sequence comprising the following sequence;

Ala Phe Leu Gly Asp Ile Ala Leu Asp Glu Glu Asp Leu Arg Ala Phe Gln Val Gln Gln Ala Val Asp Leu Arg Arg His Thr Ala Arg Lys Ser Ser Ile Lys Ala Ala, and naturally occuring allelic sequences thereof.

2. A protein according to claim 1, further characterized by cartilage and/or bone growth stimulating activity in the in vivo rat bone formation assay at a concentration of between about 10 to about 1000 nanograms per gram of bone.

3. A protein according to claim 1, further characterized by

(a) an apparent molecular weight of 28 to 30 kd as determined by SDS-polyacrylamide gel electrophoresis;

and

(b) an isoelectric point of between about pH 8.8-9.2 as determined by isoelectric focusing.

4. A therapeutic composition for treating cartilage and/or bone defects comprising an effective amount of a protein of claim 1 in a pharmaceutically acceptable vehicle.

5. A composition according to claim 4 further comprising a matrix for delivering the protein to the site of the cartilage and/or bone defect.

6. A method for treating cartilage and/or bone defects comprising administering an effective amount of a composition of claim 4.

7. A method for treating cartilege and/or bone defects comprising administering an effective amount of a composition of claim 5.