Modified animal erythropoietin polypeptides and their uses

• US 9,156,899 B2
• Filed: 09/20/2013
• Issued: 10/13/2015
• Est. Priority Date: 09/26/2008
• Status: Expired due to Fees

• Current Assignee: Elanco US Inc. (Elanco Animal Health, Inc.), Ambrx, Inc. (Johnson & Johnson)
• Original Assignee: Eli Lilly and Company, Ambrx, Inc. (Johnson & Johnson)
• Inventors: Tian, Feng, Hays Putnam, Anna-Maria A., Song, Frank, Chu, Stephanie, Sheffer, Joseph, Barnett, Richard S., Siladi, Marc, Atkinson, Kyle, Lee, Darin, Canning, Peter C.
• Primary Examiner(s): Saoud, Christine J
• Application Number:
  US14/032,963
• Publication Number:
  US 20140011740A1
• Time in Patent Office: 753 Days
• Field of Search: None
• US Class Current: 1/1
• CPC Class Codes: A61K 38/00 Medicinal preparations containing peptides peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00
• CPC Class Codes: A61K 38/1816 Erythropoietin [EPO]
• CPC Class Codes: A61K 47/60 the organic macromolecular compound being a polyoxyalkylene oligomer, polymer or dendrimer, e.g. PEG, PPG, PEO or polyglycerol
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: A61P 7/06 Antianaemics
• CPC Class Codes: C07K 14/505 Erythropoietin [EPO]
• CPC Class Codes: C12N 15/102 Mutagenizing nucleic acids

Abstract

Modified animal erythropoietin polypeptides and uses thereof are provided.

Claims

1. A feline erythropoietin (fEPO) polypeptide comprising a non-naturally encoded amino acid, wherein the non-naturally encoded amino acid is substituted at a position selected from the group consisting of residues 49, 131, 132, and 133 of SEQ ID NO:

2, and wherein the non-naturally encoded amino acid is p-acetylphenylalanine.

2. A composition comprising the fEPO polypeptide of claim 1 and a pharmaceutically acceptable carrier.

3. The composition of claim 2, wherein the non-naturally encoded amino acid is linked to a water soluble polymer, wherein the water soluble polymer comprises a poly(ethylene glycol) moiety having a molecular weight of between about 1 and about 100 kDa.

4. The fEPO polypeptide of claim 1,

and wherein the non-naturally encoded acid is linked to a water soluble polymer.

5. The fEPO polypeptide of claim 4, wherein the water soluble polymer comprises a poly(ethylene glycol) moiety having a molecular weight of between about 1 and about 100 kDa.

6. The fEPO polypeptide of claim 5, wherein the poly(ethylene glycol) moiety has a molecular weight of between about 1 kDa and 50 kDa.

7. The fEPO polypeptide of claim 6, wherein the poly(ethylene glycol) moiety has a molecular weight of between 5 kDa and 40 kDa.

8. A method of making the fEPO polypeptide of claim 4, the method comprising contacting an isolated fEPO polypeptide comprising a non-naturally encoded amino acid substituted at a position selected from the group consisting of residues 49, 131, 132, and 133 of SEQ ID NO:

2 with a water soluble polymer comprising a moiety that reacts with the non-naturally encoded amino acid, wherein the non-naturally encoded amino acid is p-acetylphenylalanine

thereby making the fEPO polypeptide comprising the non-naturally encoded amino acid linked to the water soluble polymer.

9. The method of claim 8, wherein the water soluble polymer comprises a poly(ethylene glycol) moiety having a molecular weight of between about 1 and about 100 kDa.

10. The method of claim 9, wherein the poly(ethylene glycol) moiety has an average molecular weight of between about 1 and about 50 kDa.

11. A method of making a fEPO polypeptide comprising a non-naturally encoded amino acid, the method comprising:

culturing cells comprising a polynucleotide or polynucleotides encoding a fEPO polypeptide and comprising a selector codon at a position selected from the group consisting of residues 49, 131, 132, and 133 of SEQ ID NO;

2, an orthogonal RNA synthetase and an orthogonal tRNA under conditions to permit expression of the fEPO polypeptide comprising a non-naturally encoded amino acid, wherein the non-naturally encoded amino acid is para-acetylphenylalanine; and

purifying the fEPO polypeptide from the cells.