| PATENT NUMBER | This data is not available for free |
| PATENT GRANT DATE | 31.12.02 |
| PATENT TITLE |
Prostatic growth factor |
| PATENT ABSTRACT |
The present invention relates to human PGF polypeptides and DNA (RNA) encoding such polypeptides. Also provided is a procedure for producing such polypeptides by recombinant techniques, and antibodies and antagonist/inhibitors against such polypeptides. Also provided are methods of using such polypeptides therapeutically for treating prostate cancer, to promote tissue regeneration and to facilitate wound healing. Also provided is a diagnostic assay to detect prostate cancer and benign prostatic hyperplasia. |
| PATENT INVENTORS | This data is not available for free |
| PATENT ASSIGNEE | This data is not available for free |
| PATENT FILE DATE | July 28, 1999 |
| PATENT REFERENCES CITED |
Kingsley. Genes and Development 8: 133-146, 1994.* Massague. Cell 49: 437-438, 1987.* Paralkar et al., "Cloning and characterization of a novel member of the transforming growth factor-.beta./bone morphogenetic protein family," J. Biol. Chem. 273 (22), 13760-13767 (1998). Li et al., "Placental transforming growth factor-.beta. is a downstream mediator of the growth arrest and apoptotic response of tumor cells to DNA damage and p53 overexpression," J. Biol. Chem. 275 (26), 20127-20135 (2000). Genbank Accession No. NP.sub.- 004855, "Prostate differentiation factor; PTGF-beta [Homo sapiens]," May 26, 2001. Genbank Accession No. NM.sub.- 004864, "Homo sapiens prostate differentiation factor (PLAB), mRNA," May 26, 2001. Welsh et al., "Analysis of gene expression identifies candidate markers and pharmacological targets in prostate cancer," Cancer Res. 61:5974-5978, Aug. 15, 2001. Buckhaults et al., "Secreted and cell surface genes expressed in benign and malignant colorectal tumors," Cancer Res. 61:6996-7001, Oct. 1, 2001. Bootcov et al., "MIC-1, a novel macrophage inhibitory cytokine, is a divergent member of the TGF-.beta. superfamily," Proc. Natl. Acad. Sci. USA 94:11514-11519, Oct. 1997. Bauskin et al., "The propeptide of macrophage inhibitory cytokine (MIC-1), a TGF-.beta. superfamily member, acts as a quality control determinant for correctly folded MIC-1" EMBO J. 19(10):2212-2220, 2000. Fairlie et al., "The propeptide of the transforming growth factor-.beta. superfamily member, macrophage inhibitory cytokine-1 (MIC-1), is a multifunctional domain that can facilitate protein folding and secretion," J. Biol. Chem. 276(20):16911-16918, May 18, 2000. Lawton et al., "Identification of a novel member of the TGF-beta superfamily highly expressed in human placenta," Gene 203:17-26, 1997. Bottner et al., "Characterization of the rat, mouse, and human genes of growth/differentiation factor-15/macrophage inhibiting cytokine-1 (GDF-15/MIC-1)." Gene 237:105-111, 1999. Yokoyama-Kobayashi et al., "Human cDNA encoding a novel TGF-.beta. superfamily protein highly expressed in placenta," J. Biochem. 122:622-626, 1997. Fairlie et al., "MIC-1 is a novel TGF-.beta. superfamily cytokine associated with macrophage activation," J. Leuk. Bio. 65:2-5, Jan. 1999. Fairlie et al., "Epitope mapping of the transforming growth factor-.beta. superfamily protein, macrophage inhibitory cytokine-1 (MIC-1): Identification of at least five distinct epitope specificities," Biochem. 40:65-73, 2001. Geneseq Accession No: R07335 (1991). Anscher et al., The New England Journal of Medicine, 328(22):1592-1598 (1993).* Lin et al., Cell, 68:775-785 (1992).* George et al., Macromolecular Sequencing & Synth., 127-149 (1988). |
| PATENT PARENT CASE TEXT | This data is not available for free |
| PATENT CLAIMS |
What is claimed is: 1. An isolated protein comprising an amino acid sequence selected from the group consisting of: (a) amino acid residues 1 to 295 of SEQ ID NO:2; (b) amino acid residues 2 to 295 of SEQ ID NO:2; and (c) amino acid residues 16 to 295 of SEQ ID NO:2. 2. The isolated protein of claim 1 which comprises amino acid sequence (a). 3. The isolated protein of claim 1 which comprises amino acid sequence (b). 4. The isolated protein of claim 1 which comprises amino acid sequence (c). 5. The isolated protein of claim 1 wherein the amino acid sequence further comprises a heterologous polypeptide. 6. The protein of claim 1, wherein said isolated protein is glycosylated. 7. The protein of claim 1, wherein said isolated protein is fused to polyethylene glycol. 8. A protein produced by a method comprising: (a) culturing a host cell under conditions suitable to produce the isolated protein of claim 1, wherein the host cell comprises a polynucleotide encoding the polypeptide of claim 1; and (b) recovering the protein from the host cell culture. 9. A composition comprising the isolated protein of claim 1 and a pharmaceutically acceptable carrier. 10. An isolated protein comprising an amino acid sequence selected from the group consisting of: (a) an amino acid sequence of the full-length polypeptide encoded by the cDNA in ATCC Deposit No. 75902; and (b) an amino acid sequence of the full-length polypeptide, excluding the N-terminal methionine residue, encoded by the cDNA in ATCC Deposit No. 75902. 11. The protein of claim 10 which comprises amino acid sequence (a). 12. The protein of claim 10 which comprises amino acid sequence (b). 13. The isolated protein of claim 10 wherein the amino acid sequence further comprises a heterologous polypeptide. 14. The protein of claim 10, wherein said isolated protein is glycosylated. 15. The protein of claim 10, wherein said isolated protein is fused to polyethylene glycol. 16. A protein produced by a method comprising: (a) culturing a host cell under conditions suitable to produce the isolated protein of claim 10, wherein the host cell comprises a polynucleotide encoding the polypeptide of claim 10; and (b) recovering the protein from the host cell culture. 17. A composition comprising the isolated protein of claim 10 and a pharmaceutically acceptable carrier. 18. An isolated protein comprising an amino acid sequence consisting of amino acid residues 1 to 295 of SEQ ID NO:2, wherein the amino acid sequence has one conservative substitution. 19. The isolated protein of claim 18 wherein the amino acid sequence further comprises a heterologous polypeptide. 20. The protein of claim 18, wherein said isolated protein is glycosylated. 21. The protein of claim 18, wherein said isolated protein is fused to polyethylene glycol. 22. A protein produced by a method comprising: (a) culturing a host cell under conditions suitable to produce the isolated protein of claim 18, wherein the host cell comprises a polynucleotide encoding the polypeptide of claim 18; and (b) recovering the protein from the host cell culture. 23. A composition comprising the isolated protein of claim 18 and a pharmaceutically acceptable carrier. 24. An isolated protein comprising an amino acid sequence of the full-length polypeptide encoded by the cDNA in ATCC Deposit No. 75902, wherein the amino acid sequence has one conservative substitution. 25. The isolated protein of claim 24 wherein the amino acid sequence further comprises a heterologous polypeptide. 26. The protein of claim 24, wherein said isolated protein is glycosylated. 27. The protein of claim 24, wherein said isolated protein is fused to polyethylene glycol. 28. A protein produced by a method comprising: (a) culturing a host cell under conditions suitable to produce the isolated protein of claim 24, wherein the host cell comprises a polynucleotide encoding the polypeptide of claim 24; and (b) recovering the protein from the host cell culture. 29. A composition comprising the isolated protein of claim 24 and a pharmaceutically acceptable carrier. 30. An isolated protein comprising a recombinant PGF polypeptide expressed by a cell containing a polynucleotide encoding SEQ ID NO:2 operably associated with a regulatory sequence that controls gene expression. 31. The isolated protein of claim 30, wherein said cell is a bacterial cell. 32. The isolated protein of claim 31, wherein said bacterial cell is an E. coli cell. 33. The isolated protein of claim 30, wherein said cell is a fungal cell. 34. The isolated protein of claim 33, wherein said fungal cell is a yeast cell. 35. The isolated protein of claim 30, wherein said cell is an insect cell. 36. The isolated protein of claim 30, wherein said cell is an animal cell. 37. The isolated protein of claim 30, wherein said cell is a mammalian cell. 38. The isolated protein of claim 37, wherein said mammalian cell is a COS cell. 39. The isolated protein of claim 30, wherein the isolated protein further comprises a heterologous polypeptide. 40. The protein of claim 30, wherein said isolated protein is glycosylated. 41. The protein of claim 30, wherein said isolated protein is fused to polyethylene glycol. 42. A composition comprising the isolated protein of claim 30 and a pharmaceutically acceptable carrier. 43. An isolated protein comprising a recombinant PGF polypeptide expressed by a cell containing at least the ORF of the cDNA in ATCC Deposit No. 75902 operably associated with a regulatory sequence that controls gene expression. 44. The isolated protein of claim 43, wherein said cell is a bacterial cell. 45. The isolated protein of claim 44, wherein said bacterial cell is an E. coli cell. 46. The isolated protein of claim 43, wherein said cell is a fungal cell. 47. The isolated protein of claim 46, wherein said fungal cell is a yeast cell. 48. The isolated protein of claim 43, wherein said cell is an insect cell. 49. The isolated protein of claim 43, wherein said cell is an animal cell. 50. The isolated protein of claim 43, wherein said cell is a mammalian cell. 51. The isolated protein of claim 50, wherein said mammalian cell is a COS cell. 52. The isolated protein of claim 43, wherein the isolated protein further comprises a heterologous polypeptide. 53. The protein of claim 43, wherein said isolated protein is glycosylated. 54. The protein of claim 43, wherein said isolated protein is fused to polyethylene glycol. 55. A composition comprising the isolated protein of claim 43 and a pharmaceutically acceptable carrier. -------------------------------------------------------------------------------- |
| PATENT DESCRIPTION |
This invention relates to newly identified polynucleotides, polypeptides encoded by such polynucleotides, the use of such polynucleotides and polypeptides, as well as the production of such polynucleotides and polypeptides. More particularly, the polypeptide of the present invention is a prostatic growth factor which is sometimes hereinafter referred to as "PGF". This invention relates to a polynucleotide and polypeptide molecules which are structurally and functionally related to TGF-.beta.. The transforming growth factor-beta family of peptide growth factors includes five members, termed TGF-.beta.1 through TGF-.beta.5, all of which form homo-dimers of approximately 25 kd. The TGF-.beta. family belongs to a larger, extended super family of peptide signaling molecules that includes the Muellerian inhibiting substance (Cate, R. L. et al., Cell, 45:685-698 (1986)), decapentaplegic (Padgett, R. W. et al., Nature, 325:81-84 (1987)), bone morphogenic factors (Wozney, J. M. et al., Science, 242:1528-1534 (1988)), vg1 (Weeks, D. L., and Melton, D. A., Cell, 51:861-867 (1987)), activins (Vale, W. et al., Nature, 321:776-779 (1986)), and inhibins (Mason, A. J. et al., Nature, 318:659-663 (1985)). These factors are similar to TGF-.beta. in overall structure, but share only approximately 25% amino acid identity with the TGF-.beta. proteins and with each other. All of these molecules are thought to play an important roles in modulating growth, development and differentiation. The protein of the present invention, PGF, retains the seven cysteine residues conserved in the C-terminal, active domain of TGF-.beta.. TGF-.beta. was originally described as a factor that induced normal rat kidney fibroblasts to proliferate in soft agar in the presence of epidermal growth factor (Roberts, A. B. et al., PNAS USA, 78:5339-5343 (1981)). TGF-.beta. has subsequently been shown to exert a number of different effects in a variety of cells. For example, TGF-.beta. can inhibit the differentiation of certain cells of mesodermal origin (Florini, J. R. et al., J.Biol.Chem., 261:1659-16513 (1986)), induced the differentiation of others (Seyedine, S. M. et al., PNAS USA, 82:2267-2271 (1985)), and potently inhibit proliferation of various types of epithelial cells, (Tucker, R. F., Science, 226:705-707 (1984)). This last activity has lead to the speculation that one important physiologic role for TGF-.beta. is to maintain the repressed growth state of many types of cells. Accordingly, cells that lose the ability to respond to TGF-.beta. are more likely to exhibit uncontrolled growth and to become tumorigenic. Indeed, the cells lack certain tumors such as retinoblastomas lack detectable TGF-.beta. receptors at their cell surface and fail to respond to TGF-.beta., while their normal counterparts express self-surface receptors in their growth is potently inhibited by TGF-.beta. (Kim Chi, A. et al., Science, 240:196-198 (1988)). More specifically, TGF-.beta.1 stimulates the anchorage-independent growth of normal rat kidney fibroblasts (Robert et al., PNAS USA, 78:5339-5343 (1981)). Since then it has been shown to be a multi-functional regulator of cell growth and differentiation (Sporn et al., Science, 233:532-534 (1986)) being capable of such diverse effects of inhibiting the growth of several human cancer cell lines (Roberts et al., PNAS-USA, 82:119-123 (1985)), mouse keratinocytes, (Coffey et al., Cancer RES., 48:1596-1602 (1988)), and T and B lymphocytes (Kehrl et al., J.Exp.Med., 163:1037-1050 (1986)). It also inhibits early hematopoietic progenitor cell proliferation (Goey et al., J.Immunol., 143:877-880 (1989)), stimulates the induction of differentiation of rat, muscle mesenchymal cells and subsequent production of cartilage-specific macro molecules (Seyedine et al., J.Biol.Chem., 262:1946-1949 (1986)), causes increased synthesis and secretion of collagen (Ignotz et al., J.Biol.Chem., 261:4337-4345 (1986)), stimulates bone formation (Noda et al., Endocrinology, 124;2991-2995 (1989)), and accelerates the healing of incision wounds (Mustoe et al., Science, 237:1333-1335 (1987)). Further, TGF-.beta.1 stimulates formation of extracellular matrix molecules in the liver and lung. When levels of TGF-.beta.1 are higher than normal, formation of fiber occurs in the extracellular matrix of the liver and lung which can be fatal. High levels of TGF-.beta.1 occur due to chemotherapy and bone marrow transplant as an attempt to treat cancers, eg. breast cancer. A second protein termed TGF-.beta.2 was isolated from several sources including demineralized bone, a human prostatic adenocarcinoma cell line (Ikeda et al., Bio.Chem., 26:2406-2410 (1987)). TGF-.beta.2 shared several functional similarities with TGF-.beta.1. These proteins are now known to be members of a family of related growth modulatory proteins including TGF-.beta.3 (Ten-Dijke et al., PNAS-USA, 85:471-4719 (1988)), Muellerian inhibitory substance and the inhibins. Due to amino acid sequence homology, it is thought that the PGF polypeptide of the present invention is also a member of this family of related growth modulatory proteins. However, to date, this polypeptide has only been found by the inventors to be present in the prostate. In accordance with one aspect of the present inventions there is provided a novel mature polypeptide which is PGF, as well as biologically active and diagnostically or therapeutically useful fragments, analogs and derivatives thereof. The polypeptide of the present invention is of human origin. In accordance with another aspect of the present invention, there are provided isolated nucleic acid molecules encoding human PGF, including mRNAs, DNAS, cDNAs, genomic DNAs as well as analogs and biologically active and diagnostically or therapeutically useful fragments and derivatives thereof. In accordance with yet a further aspect of the present invention, there is provided a process for producing such polypeptide by recombinant techniques comprising culturing recombinant prokaryotic and/or eukaryotic host cells, containing a human PGF nucleic acid sequence, under conditions promoting expression of said protein and subsequent recovery of said protein. In accordance with yet a further aspect of the present invention, there is provided a process for utilizing such polypeptide, or polynucleotide encoding such polypeptide for therapeutic purposes, for example, to inhibit prostate cancer, stimulate tissue regeneration and to promote wound healing. In accordance with yet a further aspect of the present invention, there are provided antibodies against such polypeptides. In accordance with yet another aspect of the present invention, there are provided antagonists to such polypeptides, which may be used to inhibit the action of such polypeptides, for example, in the treatment of PGF-dependent tumors. In accordance with yet a further aspect of the present invention, there are also provided nucleic acid probes comprising nucleic acid molecules of sufficient length to specifically hybridize to human PGF sequences. In accordance with still another aspect of the present invention, there are provided diagnostic assays for detecting diseases related to the under-expression and over-expression of the PGF polypeptide and mutations in the nucleic acid sequences encoding such polypeptide. In accordance with yet a further aspect of the present invention, there is provided a process for utilizing such polypeptides, or polynucleotides encoding such polypeptides, for in vitro purposes related to scientific research, synthesis of DNA and manufacture of DNA vectors. These and other aspects of the present invention should be apparent to those skilled in the art from the teachings herein |
| PATENT PHOTOCOPY | Available on request |
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