Claims for Patent: 7,473,680

Summary for Patent: 7,473,680

Title:	Remodeling and glycoconjugation of peptides
Abstract:	The invention includes methods and compositions for remodeling a peptide molecule, including the addition or deletion of one or more glycosyl groups to a peptide, and/or the addition of a modifying group a peptide.
Inventor(s):	DeFrees; Shawn (North Wales, PA), Zopf; David A. (Wayne, PA), Bayer; Robert J. (San Diego, CA), Hakes; David James (Willow Grove, PA), Bowe; Caryn (Doylestown, PA), Chen; Xi (Lansdale, PA)
Assignee:	Neose Technologies, Inc. (Horsham, PA)
Application Number:	11/183,205
Patent Claims:	1. A method of forming a covalent conjugate between a water soluble polymer and a glycosylated or non-glycosylated peptide, wherein said water soluble polymer is conjugated to said peptide via an intact glycosyl linking group interposed between and covalently linked to both said peptide and said water soluble polymer, said method comprising: contacting said peptide with a mixture comprising a nucleotide sugar covalently linked to said water soluble polymer and a glycosyltransferase for which said nucleotide sugar is a substrate under conditions sufficient to form said conjugate. 2. The method of claim 1, wherein said glycosyl linking group is covalently attached to a glycosyl residue covalently attached to said peptide. 3. The method of claim 1, wherein said glycosyl linking group is covalently attached to an amino acid residue of said peptide. 4. The method of claim 1, wherein said water soluble polymer comprises a member selected from the group consisting of a polyalkylene oxide and a polypeptide. 5. The method of claim 4, wherein said polyalkylene oxide is poly(ethylene glycol). 6. The method of claim 5, wherein said poly(ethylene glycol) has a degree of polymerization of from about 1 to about 20,000. 7. The method of claim 6, wherein said polyethylene glycol has a degree of polymerization of from about 1 to about 5,000. 8. The method of claim 7, wherein said polyethylene glycol has a degree of polymerization of from about 1 to about 1,000. 9. The method of claim 1, wherein said glycosyltransferase is selected from the group consisting of sialyltransferase, galactosyltransferase, glucosyltransferase, GalNAc transferase, GlcNAc transferase, fucosyltransferase, and mannosyltransferase. 10. The method of claim 1, wherein said glycosyltransferase is recombinantly produced. 11. The method of claim 10, wherein said glycosyltransferase is a recombinant prokaryotic enzyme. 12. The method of claim 10, wherein said glycosyltransferase is a recombinant eukaryotic enzyme. 13. The method of claim 1, wherein said nucleotide sugar is selected from the group consisting of UDP-glycoside, CMP-glycoside, and GDP-glycoside. 14. The method of claim 13, wherein said nucleotide sugar is selected from the group consisting of UDP-galactose, UDP-galactosamine, UDP-glucose, UDP-glucosamine, UDP-N-acetylgalactosamine, UDP-N-acetylglucosamine, GDP-mannose, GDP-fucose, CMP-sialic acid, CMP-NeuAc. 15. The method of claim 1, wherein said peptide is a therapeutic agent. 16. The method of claim 1, wherein said glycosylated peptide is partially deglycosylated prior to said contacting. 17. The method of claim 1, wherein said intact glycosyl linking group is a sialic acid residue. 18. The method of claim 1, wherein said method is performed in a cell-free environment. 19. The method of claim 1, wherein said covalent conjugate is isolated. 20. The method of claim 19, wherein said covalent conjugate is isolated by membrane filtration. 21. The method of claim 1, wherein said nucleotide sugar covalently linked to said water soluble polymer has the formula: ##STR00084## wherein R is said water soluble polymer or is said water soluble polymer attached to said nucleotide sugar through a linker; and i is a cation. 22. The method according to claim 21, wherein said water soluble polymer is poly(ethylene glycol). 23. The method according to claim 22, wherein said poly(ethylene glycol) is selected from linear and branched poly(ethylene glycol). 24. The method according to claim 22, wherein said nucleotide sugar has the formula: ##STR00085## wherein n is an integer from 1 to 2000. 25. The method according to claim 1, wherein said peptide is a glycosylated peptide. 26. The method according to claim 25, wherein said peptide comprises a glycosyl residue having a formula which is a member selected from: ##STR00086## wherein a, b, c, d, i, n, o, p, q, r, s, t, and u are members independently selected from 0 and 1; e, f, g, and h are members independently selected from the integers between 0 and 4; j, k, l, and m are members independently selected from the integers between 0 and 20; v, w, x, y, and z are 0; and R is said water soluble polymer. 27. The method of claim 25, further comprising: (b) prior to step (a), contacting said peptide with a sialidase under conditions appropriate to remove sialic acid from said peptide. 28. The method of claim 25, further comprising: (c) contacting the product of step (a) with a sialyltransferase and a sialic acid donor under conditions appropriate to transfer sialic acid to said product. 29. The method of claim 25, further comprising: (d) prior to step (a), contacting said peptide with a galactosidase operating synthetically under conditions appropriate to add a galactose to said peptide. 30. The method of claim 25, further comprising: (e) prior to step (a), contacting said peptide with a galactosyl transferase and a galactose donor under conditions appropriate to transfer said galactose to said peptide. 31. The method of claim 30, further comprising: (f) contacting the product from step (e) with ST3Gal3 and a sialic acid donor under conditions appropriate to transfer sialic acid to said product. 32. The method of claim 25, further comprising: (g) contacting the product from step (a) with a moiety that reacts with said modifying group, thereby forming a conjugate between said intact glycosyl linking group and said moiety. 33. The method of claim 25, further comprising: (h) prior to step (a), contacting said peptide with N-acetylglucosamine transferase and a GlcNAc donor under conditions appropriate to transfer GlcNAc to said peptide. 34. The method of claim 26, wherein a, b, c, d, e, f, g, n, and q are 1; h is a member selected from the integers between 1 and 3; i, j, k, l, m, o, p, r, s, t, and u are members independently selected from 0 and 1; and v, w, x, y and z are 0. 35. The method of claim 25, wherein a, b, c, d, f, h, j, k, l, m, q, s, u, v, w, x, y, and z are 0; and e, g, i, r, and t are members independently selected from 0 and 1. 36. The method of claim 26, wherein a, b, c, d, e, f, g, h, i, j, k, l, m, n, o, p, q, r, s, t, and u are members independently selected from 0 and 1; and v, w, x, y, and z are 0. 37. The method of claim 26, wherein a, b, c, d, e, f, g, n, and q are 1; h is a member selected from the integers between 1 and 3; i, j, k, l, m, o, p, r, s, t, and u are members independently selected from 0 and 1; and v, w, x, y and z are 0. 38. The method of claim 26, wherein a, b, c, d, f, h, j, k, l, m, o, p, s, u, v, w, x, y, and z are 0; and e, g, i, n, q, r, and t are independently selected from 0 and 1. 39. The method of claim 26, wherein a, b, c, d, f, h, j, k, l, m, n, o, p, s, u, v, w, x, y, and z are 0; and e, g, i, q, r, and t are members independently selected from 0 and 1. 40. The method of claim 26, wherein q is 1; a, b, c, d, e, f, g, h, i, n, r, s, t, and u are members independently selected from 0 and 1; and j, k, l, m, o, p, v, w, x, y, and z are 0. 41. The method of claim 1, wherein said peptide is selected from the group consisting of granulocyte colony stimulating factor, interferon-alpha, interferon-beta, Factor VIIa, Factor IX, follicle stimulating hormone, erythropoietin, granulocyte macrophage colony stimulating factor, interferon-gamma, alpha-1-protease inhibitor, beta-glucosidase, tissue plasminogen activator protein, interleukin-2, Factor VIII, chimeric tumor necrosis factor receptor, urokinase, chimeric anti-glycoprotein IIb/IIIa antibody, chimenc anti-HER2 antibody, chimeric anti-respiratory syncytial virus antibody, chimeric anti-CD20 antibody, DNase, chimeric anti-tumor necrosis factor antibody, human insulin, hepatitis B sAg, and human growth hormone. 42. The method of claim 1, wherein said water soluble polymer is poly(ethylene glycol) and said intact glycosyl linking group is a sialic acid residue of the formula: ##STR00087## in which R is a member selected from said poly(ethylene glycol) and said poly(ethylene glycol) attached to said sialic acid residue through said linker. 43. The method of claim 42, wherein said peptide is selected from the group consisting of granulocyte colony stimulating factor, interferon-alpha, interferon-beta, Factor VIIa, Factor IX, follicle stimulating hormone, erythropoietin, granulocyte macrophage colony stimulating factor, interferon-gamma, alpha-1-protease inhibitor, beta-glucosidase, tissue plasminogen activator protein, interleukin-2, Factor VIII, chimeric tumor necrosis factor receptor, urokinase, chimeric anti-glycoprotein IIb/IIIa antibody, chimeric anti-HER2 antibody, chimeric anti-respiratory syncytial virus antibody, chimeric anti-CD20 antibody, DNase, chimeric anti-tumor necrosis factor antibody, human insulin, hepatitis B sAg, and human growth hormone.

Details for Patent 7,473,680

Applicant	Tradename	Biologic Ingredient	Dosage Form	BLA	Approval Date	Patent No.	Expiredate
Microbix Biosystems Inc.	KINLYTIC	urokinase	For Injection	021846	January 16, 1978		2021-11-28
Novo Nordisk Inc.	REBINYN	coagulation factor ix (recombinant), glycopegylated	For Injection	125611	May 31, 2017		2021-11-28
Novo Nordisk Inc.	REBINYN	coagulation factor ix (recombinant), glycopegylated	For Injection	125611	August 11, 2022		2021-11-28