DNA molecules encoding bone morpogenetic protein-11

• US 5,639,638 A
• Filed: 05/20/1994
• Issued: 06/17/1997
• Est. Priority Date: 05/12/1993
• Status: Expired due to Term

• Current Assignee: Genetics Institute LLC (Pfizer Inc.)
• Original Assignee: Genetics Institute LLC (Pfizer Inc.)
• Inventors: Wozney, John M., Celeste, Anthony J.
• Primary Examiner(s): Walsh, Stephen G.
• Application Number:
  US08/247,907
• Time in Patent Office: 1,124 Days
• Field of Search: 536/23.4, 536/23.5, 536/23.51, 514/8, 514/12, 530/350, 530/399, 930/120, 435/69.4, 435/240.2, 435/252.3, 435/320.1
• US Class Current: 435/69.4
• CPC Class Codes: A61K 38/1875 Bone morphogenic factor; Osteogenins; Osteogenic factor; Bone-inducing factor
• CPC Class Codes: A61P 13/02 of urine or of the urinary tract, e.g. urine acidifiers
• CPC Class Codes: A61P 15/00 Drugs for genital or sexual disorders for disorders of sex hormones A61P5/24; Contraceptives
• CPC Class Codes: A61P 15/18 Feminine contraceptives
• CPC Class Codes: A61P 19/00 Drugs for skeletal disorders
• CPC Class Codes: A61P 25/00 Drugs for disorders of the nervous system
• CPC Class Codes: A61P 25/28 for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: A61P 7/00 Drugs for disorders of the blood or the extracellular fluid
• CPC Class Codes: C07K 14/51 Bone morphogenetic factor; Osteogenins; Osteogenic factor; Bone-inducing factor
• CPC Class Codes: C07K 14/575 Hormones derived from pro-opiomelanocortin, pro-enkephalin or pro-dynorphin C07K14/665, e.g. corticotropin C07K14/695
• CPC Class Codes: C07K 2319/00 Fusion polypeptide
• CPC Class Codes: C12N 2501/155 Bone morphogenic proteins [BMP]; Osteogenins; Osteogenic factor; Bone inducing factor
• CPC Class Codes: C12N 5/0619 Neurons
• 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 Bone Morphogenetic Protein-11 proteins and processes for producing them are disclosed. Recombinant DNA molecules encoding the BMP-11 proteins are also disclosed. The proteins may be useful in regulating follicle stimulating hormone, such as for contraception, and for the induction of bone, cartilage and/or other connective tissue.

Claims

1. An isolated DNA molecule comprising a DNA sequence selected from the group consisting of:

(a) nucleotide #375 or #390 to #704 of SEQ ID NO;

1;

(b) nucleotides #760 or #775 to #1086 of SEQ ID NO;

10;

(c) nucleotides encoding amino acids 1 or 6 to 109 of SEQ ID NO;

2;

(d) nucleotides encoding amino acids 1 or 6 to 109 of SEQ ID NO;

11; and

(e) naturally occurring allelic sequences and equivalent degenerative codon sequences of (a) or (b).

2. A host cell transformed with a DNA molecule of claim 1.

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

4. A host cell transformed with the vector of claim 3.

8. A method for producing a purified bone morphogenetic protein-11 (BMP-11) protein, said method comprising the steps of:

(a) culturing a host cell transformed with a DNA molecule according to claim 1; and

(b) recovering and purifying said BMP-11 protein from the culture medium.

9. The method of claim 8, wherein said host cell is transformed with a DNA molecule comprising a DNA coding sequence selected from the group consisting of:

(a) nucleotide #375 to #704 of SEQ ID NO;

1;

(b) nucleotide #760 to #1086 of SEQ ID NO;

10; and

(c) naturally occurring allelic sequences and equivalent degenerative codon sequences of (a) and (b).

10. The method of claim 9, wherein said host cell is a mammalian cell and the DNA molecule further comprises a DNA sequence encoding a propeptide from a member of the TGF-β

superfamily of proteins, said DNA sequence encoding a propeptide being linked in proper reading frame to the DNA coding sequence.

5. An isolated DNA molecule comprising a DNA sequence selected from the group consisting of nucleotides #375 to #704 of SEQ ID NO:

1 and nucleotides #760 to #1086 of SEQ ID NO;

10.

6. A vector comprising a DNA molecule of claim 5 in operative association with an expression control sequence therefor.

7. A host cell transformed with the vector of claim 6.

11. A purified bone morphogenetic protein-11 (BMP-11) polypeptide comprising the amino acid sequence from amino acid #1 to #109 as set forth in SEQ ID NO:

2.

12. A purified bone morphogenetic protein-11 (BMP-11) polypeptide comprising the amino acid sequence from amino acid #1 to #109 as set forth in SEQ ID NO:

11.

13. A purified BMP-11 polypeptide of claim 12 wherein said polypeptide is a dimer wherein each subunit comprises the amino acid sequence from amino acid #1 to #109 of SEQ ID NO:

11.

16. A purified bone morphogenetic protein-11 (BMP-11) polypeptide according to claim 12, wherein said polypeptide is a dimer comprising two subunits, wherein one subunit comprises the amino acid sequence from amino acid #1 to amino acid #109 of SEQ ID NO:

11, and one subunit comprises an amino acid sequence for a bone morphogenetic protein selected from the group consisting of BMP-1, BMP-2, BMP-3, BMP-4, BMP-5, BMP-6, BMP-7, BMP-8, BMP-9 and BMP-10.

14. A purified bone morphogenetic protein-11 (BMP-11) polypeptide produced by the steps of

(a) culturing a cell transformed with a DNA molecule comprising the nucleotide sequence from nucleotide #375 to #704 as shown in SEQ ID NO:

1; and

(b) recovering and purifying from said culture medium a polypeptide comprising the amino acid sequence from amino acid #1 to amino acid #109 of SEQ ID NO;

2.

15. A purified bone morphogenetic protein-11 (BMP-11) polypeptide produced by the steps of

(a) culturing a cell transformed with a DNA molecule comprising the nucleotide sequence from nucleotide #760 to #1086 as shown in SEQ ID NO:

10; and

(b) recovering and purifying from said culture medium a polypeptide comprising the amino acid sequence from amino acid #1 to amino acid #109 as shown in SEQ ID NO;

11.

17. A chimeric DNA molecule comprising a DNA sequence encoding a propeptide from a member of the TGF-β

superfamily of proteins linked in frame to a DNA sequence encoding a bone morphogenetic protein-11 (BMP-11) polypeptide, said BMP-11 polypeptide being selected from the group comprising amino acid #1 to #109 as set forth in SEQ ID NO;

2 and amino acid #1 to #109 as set forth in SEQ ID NO;

11.