Claims for Patent: 6,420,531

Summary for Patent: 6,420,531

Title:	Epithelial cell specific growth factor, keratinocyte growth factor (KGF)
Abstract:	Discoveries are disclosed that show particular aspects of recombinant DNA technology can be used sucessfully to produce hitherto unknown human keratinocyte growth factor (KGF) protein free of other polypeptides. These proteins can be produced in various functional forms from spontaneously secreting cells or from DNA segments introduced into cells. These forms variously enable biochemical and functional studies of this novel protein as well as production of antibodies. Means are described for determining the level of expression of genes for the KGF protein, for example, by measuring mRNA levels in cells or by measuring antigen secreted in extracellular or body fluids.
Inventor(s):	Rubin; Jeffrey S. (Rockville, MD), Finch; Paul W. (Bethesda, MD), Aaronson; Stuart A. (Great Falls, VA)
Assignee:	The United States as represented by the Department of Health and Human (N/A) (Washington, DC)
Application Number:	08/455,620
Patent Claims:	1. An isolated keratinocyte growth factor (KGF) polypeptide prepared by expressing in a host cell a DNA encoding a polypeptide having a sequence comprising amino acids 32-194 of FIG. 7. 2. The polypeptide of claim 1, wherein said DNA encodes a Met at the amino terminus. 3. The polypeptide of claim 1, wherein said host cell is selected from the group consisting of a bacterial cell, a fungal cell, a mammalian cell and an insect cell. 4. The polypeptide of claim 3, wherein said cell is a bacterial cell. 5. The polypeptide of claim 3, wherein said cell is a mammalian cell. 6. An isolated keratinocyte growth factor (KGF) polypeptide, wherein said polypeptide comprises amino acids 32 to 194 of FIG. 7 or comprises a segment of said polypeptide, wherein said polypeptide and segment thereof has mitogenic activity on BALB/MK cells. 7. The polypeptide or segment thereof of claim 6, which comprises Met at the amino terminus. 8. The polypeptide or segment thereof of claim 6, wherein five nanomolar concentration of said polypeptide or segment thereof elicits less than one-fold stimulation over background in NIH/3T3 cells. 9. The polypeptide or segment thereof of claim 6, wherein said polypeptide or segment thereof is capable of stimulating DNA synthesis in quiescent BALB/MK epidermal keratinocytes at a concentration of 0.1 nM. 10. The polypeptide or segment thereof of claim 6, wherein an amount of said polypeptide that stimulates maximal thymidine incorporation in BALB/MK keratinocyte cells, stimulates less than one-fold stimulation over background in NIH/3T3 fibroblasts. 11. The polypeptide or segment thereof of claim 6, wherein an amount of said polypeptide or segment thereof that stimulates maximal thymidine incorporation in BALB/MK keratinocyte cells, stimulates less than 1/50th of the maximal thymidine incorporation in NIH/3T3 cells stimulated by aFGF or bFGF. 12. The polypeptide or segment thereof of claim 6, wherein an amount of said polypeptide or segment thereof that stimulates maximal thymidine incorporation in BALB/MK keratinocyte cells, stimulates less than 1/10th of the maximal thymidine incorporation in NIH/3T3 fibroblasts stimulated by EGF or TGF-alpha. 13. The polypeptide or segment thereof of claim 6, wherein the maximal thymidine incorporation in BALB/MK keratinocytes stimulated by said polypeptide or segment thereof obtained within the concentration range of 0.1 to 3 nanomolar is at least twice that obtained with bFGF within the same concentration range. 14. An isolated keratinocyte growth factor (KGF) polypeptide, wherein said polypeptide comprises the amino acids 32-194 of FIG. 7 or comprises a segment of said polypeptide which is that part of the amino acid sequence of FIG. 7 that remains after the amino acid sequence of FIG. 7 is truncated from an N terminus to C terminus direction, while the region of amino acids 32-78. 15. The polypeptide or segment thereof of claim 14, which comprises Met at the amino terminus. 16. The polypeptide of claim 14 or segment thereof, which has mitogenic activity on BALB/MK keratinocyte cells. 17. The polypeptide or segment thereof of claim 14, which has mitogenic activity on epithelial cells. 18. An isolated keratinocyte growth factor (KGF) polypeptide, wherein said polypeptide comprises amino acids 32-194 of FIG. 7 or comprises a segment of said polypeptide which is that part of the amino acid sequence of FIG. 7 that remains after the amino acid sequence of FIG. 7 is truncated from the C terminus toward the N terminus, within the region of amino acids 194 to 189. 19. The polypeptide or segment thereof of claim 18, which comprises Met at the amino terminus. 20. The polypeptide or segment thereof of claim 18, which has mitogenic activity on BALB/MK keratinocyte cells. 21. The polypeptide or segment thereof of claim 18, which has mitogenic activity on epithelial cells. 22. An isolated keratinocyte growth factor (KGF) polypeptide, wherein said polypeptide comprises amino acids 32-194 of FIG. 7 or comprises a segment of said polypeptide which is that part of the amino acid sequence of FIG. 7 that remains after the amino acid sequences of FIG. 7 is truncated from an N terminus to C terminus direction, within the region of amino acids 32-78 and is truncated from the C terminus toward the N terminus, within the region of amino acids 194 to 189. 23. The polypeptide or segment thereof of claim 22, which comprises Met at the amino terminus. 24. The polypeptide or segment thereof of claim 22, which has mitogenic activity on BALB/MK keratinocyte cells. 25. The polypeptide or segment thereof of claim 22, which has mitogenic activity on epithelial cells. 26. An isolated keratinocyte growth factor (KGF) polypeptide, wherein said polypeptide comprises amino acids 32-194 of FIG. 7. 27. The polypeptide of claim 26, which comprises Met at the amino terminus. 28. An isolated keratinocyte growth factor (KGF) polypeptide prepared by expressing in a host cell a DNA encoding an amino acid sequence comprising amino acids 32-194 of FIG. 7 or encoding an amino acid sequence which is a segment of amino acids 32-194 of FIG. 7, wherein the segment is that part of the amino acid sequence of FIG. 7 that remains after the amino acid sequence of FIG. 7 is truncated from an N terminus to C terminus direction, within the region of amino acids 32-78. 29. The polypeptide or segment thereof of claim 28, wherein said DNA encodes Met at the amino terminus. 30. The polypeptide or segment thereof of claim 28, which has mitogenic activity on BALB/MK keratinocyte cells. 31. The polypeptide or segment thereof of claim 28, which has mitogenic activity on epithelial cells. 32. An isolated keratinocyte growth factor (KGF) polypeptide, wherein said polypeptide comprises amino acids 32 to 194 of FIG. 7 or comprises a segment of said polypeptide, and wherein said polypeptide and segment thereof has mitogenic activity on epithelial cells. 33. The polypeptide or segment thereof of claim 32, which comprises Met at the amino terminus. 34. The polypeptide of claim 32, that is a segment of the polypeptide of FIG. 7. 35. The polypeptide or segment thereof of claim 32, wherein five nanomolar concentration of said polypeptide or segment thereof elicits less than one-fold stimulation over background in NIH/3T3 cells. 36. The polypeptide or segment thereof of claim 32, wherein said KGF is capable of stimulating DNA synthesis in quiescent BALB/MK epidermal keratinocytes at a concentration of 0.1 nM. 37. The polypeptide or segment thereof of claim 32, wherein an amount of said polypeptide or segment thereof that stimulates maximal thymidine incorporation in BALB/MK keratinocyte cells, stimulates less than one-fold stimulation over background in NIH/3T3 fibroblasts. 38. The polypeptide or segment thereof of claim 32, wherein an amount of said polypeptide or segment thereof that stimulates maximal thymidine incorporation in a BALB/MK keratinocyte cells, stimulates less than 1/50'th of the maximal thymidine incorporation in NIH/3T3 cells stimulated by aFGF or bFGF. 39. The polypeptide or segment thereof of claim 32, wherein an amount of said polypeptide or segment thereof that stimulates maximal thymidine incorporation in BALB/MK keratinocyte cells, stimulated less than 1/10th of the maximal thymidine incorporation in NIH/3T3 fibroblasts stimulated by EGF or TGF-alpha. 40. The polypeptide or segment thereof of claim 32, wherein the maximal thymidine incorporation in BALB/MK keratinocytes stimulated by said polypeptide or segment thereof obtained within the concentration range of 0.1 to 3 nanomolar is at least twice that obtained with bFGF within the same concentration range. 41. An isolated keratinocyte growth factor (KGF) polypeptide comprising a segment of amino acids 32-194 of FIG. 7, wherein said polypeptide and segment thereof has mitogenic activity on epithelial cells and wherein said polypeptide is unglycosylated. 42. An isolated keratinocyte growth factor (KGF) polypeptide comprising a segment of amino acids 32-194 of FIG. 7, wherein the segment is that part of the amino acid sequence of FIG. 7 that remains after the amino acid sequence of FIG. 7 is truncated from an N terminus to C terminus direction, within the region of amino acids 32-78, and wherein said polypeptide is unglycosylated. 43. The polypeptide or segment thereof of any of claims 1 to 40, wherein said polypeptide or segment thereof is unglycosylated. 44. The polypeptide or segment thereof of any of claims 1 to 40, wherein said polypeptide or segment thereof is glycosylated.

Details for Patent 6,420,531

Applicant	Tradename	Biologic Ingredient	Dosage Form	BLA	Approval Date	Patent No.	Expiredate
Swedish Orphan Biovitrum Ab (publ)	KEPIVANCE	palifermin	For Injection	125103	12/15/2004		2019-07-16

International Patent Family for US Patent 6,420,531

Country	Patent Number	Estimated Expiration
Austria	197800
Australia	5049690
Australia	647732