Chemistry: molecular biology and microbiology – Micro-organism – tissue cell culture or enzyme using process... – Recombinant dna technique included in method of making a...
Reexamination Certificate
1994-06-06
2001-09-11
Romeo, David (Department: 1647)
Chemistry: molecular biology and microbiology
Micro-organism, tissue cell culture or enzyme using process...
Recombinant dna technique included in method of making a...
C435S320100, C435S252300, C435S325000, C530S399000, C530S840000, C930S120000, C514S012200, C424S423000, C424S426000, C424S484000, C536S024310, C536S023510
Reexamination Certificate
active
06287816
ABSTRACT:
The present invention relates to a novel family of purified proteins designated BMP-9 proteins and processes for obtaining them. These proteins may be used to induce bone and/or cartilage formation and in wound healing and tissue repair.
The murine BMP-9 DNA sequence (SEQ ID NO:1) and amino acid sequence (SEQ ID NO:2) are set forth in FIG.
1
. Human BMP-9 sequence is set forth in
FIG. 3
(SEQ ID NO:8 and SEQ ID NO:9). It is contemplated that BMP-9 proteins are capable of inducing the formation of cartilage and/or bone. BMP-9 proteins may be further characterized by the ability to demonstrate cartilage and/or bone formation activity in the rat bone formation assay described below.
Murine BMP-9 is characterized by comprising amino acid #319 to #428 of
FIG. 1
(SEQ ID NO:2 amino acids #1-110). Murine BMP-9 may be produced by culturing a cell transformed with a DNA sequence comprising nucleotide #610 to nucleotide #1893 as shown in
FIG. 1
(SEQ ID NO:1) and recovering and purifying from the culture medium a protein characterized by the amino acid sequence comprising amino acid #319 to #428 as shown in
FIG. 1
(SEQ ID NO:2) substantially free from other proteinaceous materials with which it is co-produced.
Human BMP-9 is expected to be homologous to murine BMP-9 and is characterized by comprising amino acid #1 (Ser, Ala, Gly) to #110 of
FIG. 3
(SEQ ID NO:9) (Arg). The invention includes methods for obtaining the DNA sequences encoding human BMP-9. This method entails utilizing the murine BMP-9 nucleotide sequence or portions thereof to design probes to screen libraries for the human gene or fragments thereof using standard techniques. Human BMP-9 may be produced by culturing a cell transformed with the BMP-9 DNA sequence and recovering and purifying BMP-9 from the culture medium. The expressed protein is isolated, recovered, and purified from the culture medium. The purified expressed protein is substantially free from other proteinaceous materials with which it is co-produced, as well as from other contaminants. The recovered purified protein is contemplated to exhibit cartilage and/or bone formation activity. The proteins of the invention may be further characterized by the ability to demonstrate cartilage and/or bone formation activity in the rat bone formation assay described below.
Human BMP-9 may be produced by culturing a cell transformed with a DNA sequence comprising nucleotide #124 to #453 as shown in SEQ ID NO:8 and recovering and purifying from the culture medium a protein characterized by the amino acid sequence of SEQ ID NO:9 from amino acid #1 to amino acid #110 substantially free from other proteinaceous materials with which it is co-produced.
Another aspect of the invention provides pharmaceutical compositions containing a therapeutically effective amount of a BMP-9 protein in a pharmaceutically acceptable vehicle or carrier. BMP-9 compositions of the invention may be used in the formation of cartilage. These compositions may further be utilized for the formation of bone. BMP-9 compositions may also be used for wound healing and tissue repair. Compositions of the invention may further include at least one other therapeutically useful agent such as the BMP proteins BMP-1, BMP-2, BMP-3, BMP-4, BMP-5, BMP-6, and BMP-7 disclosed for instance in PCT Publication Nos. WO88/00205, WO89/10409, and WO90/11366, and BMP-8, disclosed in U.S. application Ser. No. 07/641,204 filed Jan. 15, 1991, now abandoned Ser. No. 07/525,357 filed May 16, 1990, now abandoned and Ser. No. 07/800,364, U.S. Pat. No. 5,688,678, filed Nov. 20, 1991.
The compositions of the invention may comprise, in addition to a BMP-9 protein, other therapeutically useful agents including growth factors such as epidermal growth factor (EGF), fibroblast growth factor (FGF), transforming growth factor (TGF-&agr; and TGF-&bgr;), and insulin-like growth factor (IGF). The compositions may also include an appropriate matrix for instance, for supporting the composition and providing a surface for bone and/or cartilage growth. The matrix may provide slow release of the osteoinductive protein and/or the appropriate environment for presentation thereof.
The BMP-9 compositions may be employed in methods for treating a number of bone and/or cartilage defects, periodontal disease and various types of wounds. These methods, according to the invention, entail administering to a patient needing such bone and/or cartilage formation wound healing or tissue repair, an effective amount of a BMP-9 protein. These methods may also entail the administration of a protein of the invention in conjunction with at least one of the novel BMP proteins disclosed in the co-owned applications described above. In addition, these methods may also include the administration of a BMP-9 protein with other growth factors including EGF, FGF, TGF-&agr;, TGF-&bgr;, and IGF.
Still a further aspect of the invention are DNA sequences coding for expression of a BMP-9 protein. Such sequences include the sequence of nucleotides in a 5′ to 3′ direction illustrated in
FIG. 1
(SEQ ID NO:1) and
FIG. 3
(SEQ ID NO:8) or DNA sequences which hybridize under stringent conditions with the DNA sequences of
FIG. 1
or
3
and encode a protein having the ability to induce the formation of cartilage and/or bone. Finally, allelic or other variations of the sequences of
FIG. 1
or
3
, whether such nucleotide changes result in changes in the peptide sequence or not, are also included in the present invention.
A further aspect of the invention includes vectors comprising a DNA sequence as described above in operative association with an expression control sequence therefor. These vectors may be employed in a novel process for producing a BMP-9 protein of the invention in which a cell line transformed with a DNA sequence encoding a BMP-9 protein in operative association with an expression control sequence therefor, is cultured in a suitable culture medium and a BMP-9 protein is recovered and purified therefrom. This process may employ a number of known cells both prokaryotic and eukaryotic as host cells for expression of the polypeptide.
Other aspects and advantages of the present invention will be apparent upon consideration of the following detailed description and preferred embodiments thereof.
REFERENCES:
patent: 4294753 (1981-10-01), Urist
patent: 4434094 (1984-02-01), Seyedin et al.
patent: 4455256 (1984-06-01), Urist
patent: 4563350 (1986-01-01), Nathan
patent: 4608199 (1986-08-01), Caplan et al.
patent: 4619989 (1986-10-01), Urist
patent: 4627982 (1986-12-01), Seyedin et al.
patent: 4681763 (1987-07-01), Nathanson
patent: 4737578 (1988-04-01), Evans
patent: 4761471 (1988-08-01), Urist
patent: 4774228 (1988-09-01), Seyedin
patent: 4774322 (1988-09-01), Seyedin
patent: 4789732 (1988-12-01), Urist
patent: 4795804 (1989-01-01), Urist
patent: 4798885 (1989-01-01), Mason
patent: 4804744 (1989-02-01), Sen
patent: 4810691 (1989-03-01), Seyedin
patent: 4843063 (1989-06-01), Seyedin
patent: 4886747 (1989-12-01), Derynck
patent: 4968590 (1990-11-01), Kuberasampath et al.
patent: 5011691 (1991-04-01), Oppermann
patent: 5106626 (1992-04-01), Parsons et al.
patent: 5108753 (1992-04-01), Kuberasampath
patent: 5168050 (1992-12-01), Hammonds, Jr. et al.
patent: 5661007 (1997-08-01), Wozney et al.
patent: 2 017466 (1990-05-01), None
patent: 33 6760 (1989-06-01), None
patent: 4 165 78A2 (1990-05-01), None
patent: 4 094 72 A1 (1990-11-01), None
patent: WO 89/09788 (1989-10-01), None
patent: WO 89/09787 (1989-10-01), None
patent: WO 90/03733 (1990-04-01), None
patent: WO 91/02744 (1991-03-01), None
patent: WO 91/05802 (1991-05-01), None
patent: WO 91/18047 (1991-11-01), None
patent: WO 92/07073 (1992-04-01), None
patent: WO 92/07004 (1992-04-01), None
patent: WO 93/04692 (1993-03-01), None
patent: WO 93/05751 (1993-04-01), None
Sporn et al. 1988. Peptide growth factors are multifunctional. Nature, vol. 332, pp. 217-219, Mar. 1988.*
Baird et al. 1986. Inhbibition of endothelial cell proliferation by type-beta tran
Celeste Anthony J.
Rosen Vicki A.
Song Jeffrey R.
Thies R. Scott
Wozney John M.
Genetics Institute Inc.
Kapinos Ellen J.
Romeo David
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