Methods and compositions for the prevention and treatment of anemia

• US 7,973,009 B2
• Filed: 10/26/2007
• Issued: 07/05/2011
• Est. Priority Date: 10/23/1998
• Status: Expired due to Fees

• Current Assignee: Amgen Inc.
• Original Assignee: Amgen Inc.
• Inventors: Egrie, Joan C., Elliott, Steven G., Sitney, Karen C., Browne, Jeffrey K.
• Primary Examiner(s): Allen; Marianne P
• Assistant Examiner(s): DeBerry; Regina M
• Application Number:
  US11/977,845
• Publication Number:
  US 20090017137A1
• Time in Patent Office: 1,348 Days
• Field of Search: None
• US Class Current: 514/7.7
• CPC Class Codes: A61P 7/06 Antianaemics
• CPC Class Codes: C07K 14/505 Erythropoietin [EPO]
• CPC Class Codes: C07K 2319/30 Non-immunoglobulin-derived peptide or protein having an immunoglobulin constant or Fc region, or a fragment thereof, attached thereto
• CPC Class Codes: C12N 15/62 DNA sequences coding for fusion proteins

Abstract

Methods for increasing and maintaining hematocrit in a mammal comprising administering a hyperglycosylated analog of erythropoietin are disclosed. An analog may be administered less frequently than an equivalent molar amount of recombinant human erythropoietin to obtain a comparable target hematocrit and treat anemia. Alternatively, a lower molar amount of a hyperglycosylated analog may be administered to obtain a comparable target hematocrit and treat anemia. Also disclosed are new hyperglycosylated erythropoietin analogs, methods of production of the analogs, and compositions comprising the analogs.

Claims

1. A method of raising and maintaining hematocrit in a mammal comprising administering an amount of an analog of human erythropoietin in a pharmaceutical composition effective to raise and maintain hematocrit, wherein the analog comprises the amino acid sequence of human erythropoietin from residues 1-165 as shown in SEQ ID NO:

1 except for one or more amino acid changes which provide for one or more additional glycosylation site(s) as compared to human erythropoietin, wherein one additional site is introduced at position 52, 53, 55, 86 or 114 and an N-linked carbohydrate chain is attached at said one additional site.

2. The method of claim 1 wherein the analog comprises three, four, or more than four additional glycosylation sites wherein a carbohydrate chain is attached at each said additional site.

3. The method of claim 1 wherein additional glycosylation sites are introduced at positions 30 and 55 and N-linked carbohydrate chains are attached at the additional sites.

4. The method of claim 1 wherein additional glycosylation sites are introduced at positions 30 and 114 and N-linked carbohydrate chains are attached at the additional sites.

5. The method of claim 1 wherein additional glycosylation sites are introduced at positions 30, 55 and 88 and N-linked carbohydrate chains are attached at the additional sites.

6. The method of claim 1 wherein additional glycosylation sites are introduced at positions 30, 55 and 114 and N-linked carbohydrate chains are attached at the additional sites.

7. The method of claim 1 wherein additional glycosylation sites are introduced at positions 30, 88 and 114 and N-linked carbohydrate chains are attached at the additional sites.

8. The method of claim 1 wherein additional glycosylation sites are introduced at positions 30, 55, 88 and 114 and an N-linked carbohydrate chain is attached at the additional sites.

9. The method of claim 1 further comprising an additional glycosylation site at position 125 and wherein an O-linked carbohydrate chain is attached at the additional site.

10. The method of claim 1 wherein the analog comprises the amino acid sequence from residues 1-165 as shown in SEQ ID NO:

1 except for Asn at position 30, Thr at position 32, Asn at position 55, Thr at position 57, Val at position 87, Asn at position 88, Thr at position 90, Asn at position 114, Thr at position 116, and any one or more of Pro at position 124, Thr at position 125 and Thr at position 126.

11. The method of claim 1 wherein the analog comprises the amino acid sequence from residues 1-165 as shown in SEQ ID NO:

1 except for the amino acid changes selected from the group consisting of;

12. The method of claim 1 wherein the analog is administered less frequently than an equivalent molar amount of recombinant human erythropoietin to obtain a comparable target hematocrit.

13. The method of claim 1 wherein the amount of analog is administered about one time per week, about one time every other week, or about one time per month.

14. The method of claim 1 wherein the analog is administered at a lower molar amount than recombinant human erythropoietin to obtain a comparable target hematocrit.

15. The method of claim 12 or 14 wherein the target hematocrit is at least about 30%.

16. The method of claim 1 wherein the amount of analog that is administered is about 0.025 to 1.5 μ

g erythropoietin peptide per kg per dose three times per week.

17. The method of claim 1 wherein the amount of analog that is administered is less than about 0.025 μ

g erythropoietin peptide per kg per dose three times per week.

18. The method of claim 1 wherein the mammal is being treated for anemia.

19. The method of claim 18 wherein the anemia is associated with a decline or loss of kidney function, myelosuppressive therapy, cancer, viral infection, chronic disease, or excessive loss of blood.

20. The method of claim 19 wherein the myelosuppressive therapy comprises chemotherapeutic or anti-viral drugs.

21. The method of claim 1 further comprising administering a therapeutically effective amount of iron to maintain increased erythropoiesis during therapy.