Novel BMP products

• US 20030049826A1
• Filed: 03/09/2001
• Published: 03/13/2003
• Est. Priority Date: 07/01/1986
• Status: Active Grant

• Current Assignee: Genetics Institute LLC (Pfizer Inc.)
• Original Assignee: Genetics Institute LLC (Pfizer Inc.)
• Inventors: Wozney, John M., Wang, Elizabeth A., Rosen, Vicki A.
• Granted Patent: US 7,300,772 B2
• Time in Patent Office: Days
• US Class Current: 435/226
• 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
• CPC Class Codes: Y10S 930/12 Growth hormone, growth factor other than t-cell or b-cell growth factor, and growth hormone releasing factor; related peptides

Abstract

Purified BMP-2 and BMP-4 proteins and processes for producing them are disclosed. The proteins may be used in the treatment of bone and cartilage defects and in wound healing and related tissue repair.

Claims

1. An osteogenic protein comprising an amino acid sequence selected from the group consisting of:

(a) amino acids #32-#129 of FIG. 1 (SEQ ID NO;

     2);

(b) amino acids #299-#396 of FIG. 2 (SEQ ID NO;

     4); and

(c) amino acids #311-#408 of FIG. 3 (SEQ ID NO;

     6).

9. A protein of claim 1 further characterized by the ability to induce the formation of cartilage and/or bone.

2. An osteogenic protein selected from the group consisting of:

(a) a purified BMP-2 protein produced by the steps of

(i) culturing a cell transformed with

(a) a cDNA comprising the nucleotide sequence from nucleotide #356 to #1543 as shown in FIG. 2 (SEQ ID NO;

     3); and

(b) sequences which hybridize to sequence under stringent conditions and encode a protein having cartilage and/or bone induction activity

(ii) recovering and purifying from said culture medium a protein comprising the 97 amino acid sequence from amino acid #299 to amino acid #396 as shown in FIG. 2 (SEQ ID NO;

     4); and

(b) a purified BMP-4 protein produced by the steps of

(i) culturing a cell transformed with

(a) a cDNA comprising the nucleotide sequence from nucleotide #403 to #1626 as shown in FIG. 3 (SEQ ID NO;

     5); and

(b) sequences which hybridize to sequence (a) under stringent conditions and encode a protein having cartilage and/or bone induction activity;

(ii) recovering from said culture medium a protein comprising the 97 amino acid sequence from amino acid #311 to amino acid #408 as shown in FIG. 3 (SEQ ID NO;

     6).

3. A BMP-2 protein comprising amino acids #283-#396 of FIG. 2 (SEQ ID NO:

4).

4. The protein of claim 3 wherein said protein is a disulfide linked dimer wherein at least one of the subunits of said dimer comprises amino acid #283-#396 of FIG. 2 (SEQ ID NO:

4).

5. A purified BMP-2 protein produced by the steps of

(a) culturing a cell transformed with a cDNA comprising the nucleotide sequence from nucleotide #356 to #1543 as shown in FIG. 2 (SEQ ID NO:

3); and

(b) recovering and purifying from said culture medium a protein comprising the 97 amino acid sequence from amino acid #299 to amino acid #396 as shown in FIG. 2 (SEQ ID NO;

     4).

6. A BMP-4 protein comprising amino acids #293-#408 of FIG. 3 (SEQ ID NO:

6).

7. The protein of claim 6 wherein said protein is a disulfide linked dimer wherein at least one of the subunits of said dimer comprises amino acid #293-#408 of FIG. 3 (SEQ ID NO:

6).

8. A purified BMP-4 protein produced by the steps of

(a) culturing a cell transformed with a cDNA comprising the nucleotide sequence from nucleotide #403 to #1626 as shown in FIG. 3 (SEQ ID NO:

5); and

(b) recovering form said culture medium a protein comprising the 97 amino acid sequence from amino acid #311 to amino acid #408 as shown in FIG. 3 (SEQ ID NO;

     6).

10. A pharmaceutical composition comprising an effective amount of a BMP-2 protein in admixture with a pharmaceutically acceptable vehicle.

11. A pharmaceutical composition comprising an effective amount of a BMP-4 protein in admixture with a pharmaceutically acceptable vehicle.

18. A method for inducing bone and/or cartilage formation in a patient in need of same comprising administering to said patient an effective amount of the composition of claim 11.

19. A method for inducing bone and/or cartilage formation in a patient in need of same comprising administering to said patient an effective amount of the composition of claim 11.

12. A composition for bone and/or cartilage formation comprising an effective amount of a BMP-2 protein in a pharmaceutically acceptable vehicle.

14. A composition of claim 12 further comprising a matrix for supporting said composition and providing a surface for bone and/or cartilage growth.

16. The composition of claim 14 wherein said matrix comprises a material selected from the group consisting of hydroxyapatite, collagen, polylactic acid and tricalcium phosphate.

13. A composition for bone and/or cartilage formation comprising an effective amount of a BMP-4 protein in a pharmaceutically acceptable vehicle.

15. A composition of claim 13 further comprising a matrix for supporting said composition and providing a surface for bone and/or cartilage growth.

17. The composition of claim 15 wherein said matrix comprises a material selected from the group consisting of hydroxyapatite, collagen, polylactic acid and tricalcium phosphate.

20. A pharmaceutical composition for wound healing and tissue repair said composition comprising an effective amount of a BMP-2 protein in a pharmaceutically acceptable vehicle.

21. A pharmaceutical composition for wound healing and tissue repair said composition comprising an effective amount of a BMP-4 protein in a pharmaceutically acceptable vehicle.

22. A method for treating wounds and/or tissue repair in a patient in need of same comprising administering to said patient an effective amount of the composition of claim 21.

23. A method for treating wounds and/or tissue repair in a patient in need of same comprising administering to said patient an effective amount of the composition of claim 22.