Use of transthyretin peptide/protein fusions to increase the serum half-life of pharmacologically active peptides/proteins

• US 20030191056A1
• Filed: 04/04/2002
• Published: 10/09/2003
• Est. Priority Date: 04/04/2002
• Status: Abandoned Application

• Current Assignee: Amgen Inc.
• Original Assignee: Amgen Inc.
• Inventors: Xiong, Fei, Walker, Kenneth
• Application Number:
  US10/117,109
• Publication Number:
  US 20030191056A1
• Time in Patent Office: Days
• US Class Current: 514/7.8
• CPC Class Codes: A61K 38/043 Kallidins; Bradykinins; Related peptides
• CPC Class Codes: A61K 38/10 Peptides having 12 to 20 amino acids A61K38/043 - A61K38/046 take precedence
• CPC Class Codes: A61K 38/20 Interleukins [IL]
• CPC Class Codes: A61K 38/29 Parathyroid hormone, i.e. parathormone; Parathyroid hormone-related peptides
• CPC Class Codes: A61K 47/58 obtained by reactions only involving carbon-to-carbon unsaturated bonds, e.g. poly[meth]acrylate, polyacrylamide, polystyrene, polyvinylpyrrolidone, polyvinylalcohol or polystyrene sulfonic acid resin
• 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: A61K 47/61 the organic macromolecular compound being a polysaccharide or a derivative thereof
• CPC Class Codes: A61K 47/643 Albumins, e.g. HSA, BSA, ovalbumin or a Keyhole Limpet Hemocyanin [KHL]
• CPC Class Codes: A61P 3/10 for hyperglycaemia, e.g. antidiabetics
• CPC Class Codes: A61P 5/18 of the parathyroid hormones
• CPC Class Codes: A61P 7/04 Antihaemorrhagics; Procoagulants; Haemostatic agents; Antifibrinolytic agents
• CPC Class Codes: A61P 9/12 Antihypertensives
• CPC Class Codes: C07K 14/47 from mammals
• CPC Class Codes: C07K 2319/31 fusions, other than Fc, for prolonged plasma life, e.g. albumin

Abstract

The present invention provides a means for increasing the serum half-life of a selected biologically active agent by utilizing transthyretin (TTR) as a fusion partner with a biologically active agent. Specifically, the present invention provides substantially homogenous preparations of TTR (or a TTR variant)-biologically active agent fusions and PEG-TTR (PEG-TTR variant)-biologically active agent fusions. As compared to the biologically active agent alone, the TTR-biologically active agent fusion and/or PEG-TTR-biologically active agent fusion has substantially increased serum half-life.

Claims

1. A method for increasing the serum half-life of a biologically active agent comprising fusing the biologically active agent to transthyretin (TTR) or a TTR variant.

2. The method of claim 1 where said TTR or TTR variant is chemically modified with a chemical selected from the group consisting of dextran, poly(n-vinyl pyurrolidone), polyethylene glycols, propropylene glycol homopolymers, polypropylene oxide/ethylene oxide co-polymers, polyoxyethylated polyols and polyvinyl alcohols.

3. The method of claim 2 where said TTR or TTR variant is chemically modified with polyethylene glycol.

4. The method of claim 3 wherein said polyethylene glycol has a molecular weight of between about 1 kD and 100 kD.

5. The method of claim 4 wherein said polyethylene glycol has a molecular weight of between about 5 kD and 30 kD.

6. The method of claim 1 wherein said TTR is encoded by the nucleic acid molecule of SEQ ID NO:

2.

7. The method of claim 1 wherein the biologically active agent is a protein.

8. The method of claim 1 wherein the biologically active agent is a peptide.

9. The method of claim 8 wherein the peptide is a TPO mimetic peptide (TMP).

10. A substantially homogenous preparation of a TTR-biologically active agent fusion, optionally in a pharmaceutically acceptable diluent, carrier or adjuvant.

20. The preparation of any of claims 10-19 wherein the fusion contains a linker peptide.

11. A substantially homogenous preparation of a PEG-TTR-biologically active agent fusion, optionally in a pharmaceutically acceptable diluent, carrier or adjuvant.

12. The preparation of claim 11 wherein the biologically active agent is a protein.

13. The preparation of claim 11 wherein the biologically active agent is a peptide.

14. The preparation of claim 13 wherein the peptide is a TPO mimetic peptide (TMP).

25. A method of treating thrombocytopenia comprising administering a therapeutically effective dose of a preparation of claim 14.

15. A substantially homogenous preparation of a TTR variant-biologically active agent fusion, optionally in a pharmaceutically acceptable diluent, carrier or adjuvant.

16. A substantially homogenous preparation of a PEG-TTR variant-biologically active agent fusion, optionally in a pharmaceutically acceptable diluent, carrier or adjuvant.

17. The preparation of claim 16 wherein the biologically active agent is a protein.

18. The preparation of claim 16 wherein the biologically active agent is a peptide.

19. The preparation of claim 18 wherein the peptide is a TPO mimetic peptide (TMP).

26. A method of treating thrombocytopenia comprising administering a therapeutically effective dose of a preparation of claim 19.

21. A process for preparing a substantially homogenous preparation of a TTR-biologically active agent fusion comprising:

(a) fusing said TTR to a biologically active agent to provide a TTR-biologically active agent fusion; and

(b) isolating said TTR-biologically active agent fusion.

22. A process for preparing a substantially homogenous preparation of a TTR variant-biologically active agent fusion comprising:

(a) engineering a cysteine residue into a specific amino acid position within the amino acid sequence of said TTR to provide a variant of said TTR;

(b) fusing said TTR variant to a biologically active agent to provide a TTR variant-biologically active agent fusion; and

(c) isolating said TTR variant-biologically active agent fusion.

23. A process for preparing a substantially homogenous preparation of a PEG-TTR-biologically active agent fusion comprising:

(a) conjugating a polyethylene glycol to said TTR to provide a PEG-TTR;

(b) fusing said PEG-TTR to a biologically active agent to provide a PEG-TTR-biologically active agent fusion; and

(c) isolating said PEG-TTR-biologically active agent fusion.

24. A process for preparing a substantially homogenous preparation of a PEG-TTR variant-biologically active agent fusion comprising:

(a) engineering a cysteine residue into a specific amino acid position within the amino acid sequence of said TTR to provide a variant of said TTR;

(b) conjugating a polyethylene glycol to said TTR variant at said cysteine residue to provide a PEG-TTR variant;

(c) fusing said PEG-TTR variant to a biologically active agent to provide a PEG-TTR-biologically active agent fusion; and

(d) isolating said PEG-TTR-biologically active agent fusion.