DNA sequences encoding BMP-6 proteins

• US 5,187,076 A
• Filed: 03/07/1990
• Issued: 02/16/1993
• Est. Priority Date: 07/01/1986
• Status: Expired due to Term

• Current Assignee: Genetics Institute LLC (Pfizer Inc.)
• Original Assignee: Genetics Institute LLC (Pfizer Inc.)
• Inventors: Wozney, John M., Wang, Elizabeth A., Rosen, Vicki A., Celeste, Anthony J.
• Primary Examiner(s): Ellis, Joan
• Application Number:
  US07/490,033
• Time in Patent Office: 1,077 Days
• Field of Search: 435/69.1, 435/91, 435/172.1, 435/235, 435/240.1, 435/262.3, 435/320.1, 435/172.3, 435/255, 435/256, 536/27, 510/350, 535/18, 535/22, 535/24, 535/38, 535/55, 535/62, 535/70, 535/72
• US Class Current: 435/69.1
• 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: C12N 15/85 for animal cells

Abstract

Purified BMP-6 proteins and processes for producing them are disclosed. The proteins may be used in the treatment of bone and/or cartilage defects and in wound healing and related tissue repair.

Claims

1. An isolated DNA sequence encoding a BMP-6 protein said DNA sequence comprising the nucleotide sequence selected from the group consisting of:

a) nucleotide #1 through nucleotide #666 of FIG. 2;

b) nucleotide #1393 to #1698 of FIG. 3;

c) nucleotide #1 to #2393 of FIG. 3;

d) nucleotide #160 to #1698 of FIG. 3;

e) nucleotide #1303 to #1698 FIG. 3;

f) allelic variants and degenerative sequences of (a) through (d); and

g) sequences which

(1) hybridize to said DNA sequence under stringent hybridization conditions; and

(2) encode a protein characterized by the ability to induce the formation of cartilage and/or bone in the Rose-modified Sampath-Reddi assay.

2. A vector comprising a DNA sequence of claim 1 in operative association with an expression control sequence therefor.

3. A host cell transformed with a vector of claim 2.

4. A host cell of claim 3 wherein said host cell is a mammalian cell.

5. An isolated DNA sequence encoding the BMP-6 protein of FIG. 3 or a fragment thereof which encodes a protein characterized by the ability to induce the formation of cartilage and/or bone in the Rosen-modified Sampath-Reddi assay" after "3".

6. An isolated DNA sequence encoding a BMP-6 protein said sequence comprising nucleotide #1 to #2923 of FIG. 3.

7. A vector comprising the nucleotide sequence of claim 6.

8. A host cell transformed with the vector of claim 7.

9. The host cell of claim 8 wherein said host cell is a mammalian cell.

10. A method for producing a purified BMP-6 protein said method comprising the steps of

(a) culturing in a suitable culture medium cells transformed with a DNA sequence comprising the DNA nucleotide #1 to #2923 of FIG. 3;

and

(b) isolating and purifying said protein from said culture medium said BMP-6 protein.

11. An isolated DNA sequence encoding a BMP-6 protein said sequence comprising nucleotide #160 to #1698 of FIG. 3.

12. A vector comprising the nucleotide sequence of claim 11.

13. A host cell transformed with the vector of claim 12.

14. The host cell of claim 13 wherein said host cell is a mammalian cell.

15. A method for producing a purified BMP-6 protein said method comprising the steps of

(a) culturing in a suitable culture medium cells transformed with a DNA sequence comprising the DNA nucleotide #5/8to #1698 of FIG. 3;

and

(b) isolating and purifying said protein from said culture medium said BMP-6 protein.

16. An isolated DNA sequence encoding a BMP-6 protein said sequence comprising nucleotide #1303 to #1698 of FIG. 3.

17. A vector comprising the nucleotide sequence of claim 16.

18. A host cell transformed with the vector of claim 17.

19. The host cell of claim 18 wherein said host cell is a mammalian cell.

20. A method for producing a purified BMP-6 protein said method comprising the steps of

(a) culturing in a suitable culture medium cells transformed with a DNA sequence comprising the DNA nucleotide #1303 to #1698 of FIG. 3;

and

(b) isolating and purifying said protein from said culture medium said BMP-6 protein.

21. An isolated DNA sequence encoding a BMP-6 protein said sequence comprising nucleotide #1393 to #1698 of FIG. 3.

22. A vector comprising the nucleotide sequence of claim 21.

23. A host cell transformed with the vector of claim 22.

24. The host cell of claim 23 wherein said host cell is a mammalian cell.

25. A method for producing a purified BMP-6 protein said method comprising the steps of

(a) culturing in a suitable culture medium cells transformed with a DNA sequence comprising the DNA nucleotide #1393 to #1698 of FIG. 3;

and

(b) isolating and purifying said protein form said culture medium said protein comprising amino acid #412 to #513 as shown in FIG. 3.

26. An isolated DNA sequence encoding the BMP-6 protein of FIG. 2.

27. An isolated DNA sequence encoding a BMP-6 protein comprising the nucleotide sequence from nucleotide #1 to #666 of FIG. 2.