Pharmacodynamically enhanced therapeutic proteins
Compositions containing an N-linked oligosaccharide having little or no terminal sialic acid residues which is attached to a therapeutic protein that is bonded to a glycol polymer.
1 . A therapeutic protein comprising an N-linked oligosaccharide wherein the N-linked oligosaccharide substantially excludes sialic acid and the therapeutic protein is chemically bonded to a glycol polymer.
2 . The therapeutic protein of claim 1 comprising an O-linked oligosaccharide.
3 . The therapeutic protein of claim 1 wherein two N-linked oligosaccharides exclude a sialic acid.
4 . The therapeutic protein of claim 1 wherein three N-linked oligosaccharides exclude a sialic acid.
5 . The composition of claim 1 wherein the glycol polymer is a polyalkylene glycol.
6 . The therapeutic protein of claim 1 wherein the glycol polymer is polyethylene glycol.
7 . The therapeutic protein of claim 1 wherein the glycol polymer has a molecular weight between about 300 KDa and about 200,000 KDa.
8 . The composition of claim 1 wherein the glycol polymer is covalently bonded to at least one of an amino group, a hydroxyl group, a sulfhydryl group and a carboxyl group of the therapeutic protein.
9 . The therapeutic protein of claim 1 obtained from a transgenic avian.
10 . The composition of claim 9 wherein the avian is selected from the group consisting of chicken, turkey and quail.
11 . The composition of claim 9 wherein the avian is a chicken.
12 . The composition of claim 9 wherein the therapeutic protein is produced in a tubular gland cell.
13 . The composition of claim 1 wherein the therapeutic protein excludes a terminal sialic acid.
14 . The therapeutic protein of claim 1 wherein the therapeutic protein is selected from the group consisting of EPO, Factor VIII, Factor VIIa, Factor IX, alteplase tPA, Reteplase tPA (differs from h tPA—3 of 5 domains deleted), growth hormone (e.g., hGH), thyroid stimulatin hormone (TSH), follicle stimulating hormone (FSH), follitropin-beta, calcitonin, platelet derived growth factor (PDGF), keratinocyte growth factor, insulin-like growth factor-1 (IGF-1), IGFBP-3, GM-CSF, INF-beta, INF-beta1b and IFN-gamma, IFN-gamma 1b, IL-3 and IL-12, TNFR-IgG fragment fusion protein, beta glucocerebrosidase, asparaginase, urokinase, adenosin deaminase, agalsidase alfa, idursulfase, alpha-L-iduronidase, galsulfase: arylsulfatase, galsulfase: arylsulfatase B, BM 102, N-acetylgalactosamine-4-sulfatase, hASB, activated protein C, dornase-alpha DNAse, anakinra, eptotermin alfa, Protein C and dibotermin alfa.
15 . The therapeutic protein of claim 1 wherein the therapeutic protein is erythropoietin.
16 . The composition of claim 15 wherein the erythropoietin is human erythropoietin.
17 . The therapeutic protein of claim 1 wherein the therapeutic protein has the amino acid sequence set forth in FIG. 2B .
18 . A composition comprising an asialic therapeutic protein obtained from a transgenic avian covalently bonded to a glycol polymer.
19 . The composition of claim 18 wherein the therapeutic protein is erythropoietin.
20 . The composition of claim 19 wherein the amino acid sequence of the erythropoietin is set forth in FIG. 2B .
21 . The composition of claim 18 wherein the therapeutic protein is produced in an oviduct cell of the transgenic avian.
22 . The composition of claim 21 wherein the oviduct cell is a tubular gland cell.
23 . The composition of claim 18 wherein the glycol polymer is a polyethylene glycol.
24 . The composition of claim 18 wherein the glycol polymer has a molecular weight of about 300 to about 200,000.
25 . A method comprising,
producing an therapeutic protein which excludes sialic acid being terminally linked to an oligosaccharide of the therapeutic protein; and covalently bonding a glycol polymer to the therapeutic protein.
26 . A pharmaceutical composition comprising erythropoietin according claim 25 .
27 . A method of treating a condition comprising administering to a patient in need thereof a pharmaceutical composition of claim 26 .