Fusion protein of human fibroblast growth factor-21 and exendin-4

• US 8,809,499 B2
• Filed: 03/12/2010
• Issued: 08/19/2014
• Est. Priority Date: 08/20/2009
• Status: Active Grant

• Current Assignee: Zyme Biotech Inc.
• Original Assignee: Chongqing Fagen Biomedical, Inc.
• Inventors: Fan, Kai, Zhao, Zhiquan, Zhang, Chun, Chen, Yong, Luo, Hua, Yang, Li, Liu, Honghong
• Primary Examiner(s): Saoud, Christine J
• Application Number:
  US13/391,226
• Publication Number:
  US 20120238496A1
• Time in Patent Office: 1,621 Days
• Field of Search: None
• US Class Current: 530/350
• 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: A61P 3/00 Drugs for disorders of the metabolism of the blood or the extracellular fluid A61P7/00
• CPC Class Codes: A61P 3/04 Anorexiants; Antiobesity agents
• CPC Class Codes: A61P 3/06 Antihyperlipidemics
• CPC Class Codes: A61P 3/10 for hyperglycaemia, e.g. antidiabetics
• CPC Class Codes: C07K 14/43 Sweetening agents, e.g. thaumatin, monellin
• CPC Class Codes: C07K 14/50 Fibroblast growth factor [FGF]
• CPC Class Codes: C07K 14/605 Glucagons
• CPC Class Codes: C07K 2319/00 Fusion polypeptide

Abstract

Provided is a fusion protein, which comprises human fibroblast growth factor 21 and glucagon-like-peptide-1 or its analogs. Also provided is the medicament composition comprising the fusion protein, which can be used for treating or preventing obesity, diabetes, hyperglycemia and hyperlipidemia etc.

Claims

1. A fusion protein having a formula of R2-R1 or R2-L-R1, wherein:

R1 is a human fibroblast growth factor 21;

R2 is Exendin-4; and

L is a linker.

2. The fusion protein according to claim 1, wherein the Exendin-4 has the amino acid sequence of SEQ ID NO:

3.

3. The fusion protein according to claim 1, wherein the linker is:

(a) (Gly-Gly-Gly-Gly-Ser)_n-(Ser)_m, wherein n is an integer between 1-5, and m is 0 or 1;

(b) (Gly-Gly-Gly-Gly-Gly)_n-(Ser)_m, wherein n is an integer between 1-5, and m is 0 or 1;

(c) (Gly-Gly-Gly-Gly-Ser)_n-(Ser-Pro)_m, wherein n is an integer between 1-5, and m is 0 or 1;

(d) (Pro-Glu-Ala-Pro-Thr-Asp)_n, wherein n is an integer between 1-5;

or

(e) (Ser-Ser-Ser-Ser-Gly)_n-(Ser-Pro)_m, wherein n is an integer between 1-5, and m is 0 or 1.

4. The fusion protein according to claim 1, wherein the human fibroblast growth factor 21 comprises the amino acid sequence of SEQ ID NO:

1.

5. A polynucleotide, which encodes a fusion protein,

wherein the fusion protein has a formula of R2-R1 or R2-L-R1, wherein:

R1 is a human fibroblast growth factor 21;

R2 is Exendin-4; and

L is a linker.

6. A vector, which comprises a polynucleotide,

wherein the polynucleotide encodes a fusion protein, and wherein the fusion

protein has a formula of R2-R1 or R2-L-R1, wherein:

R1 is a human fibroblast growth factor 21;

R2 is Exendin-4; and

L is a linker.

7. A host cell, which contains a vector, wherein the vector comprises a polynucleotide, wherein the polynucleotide encodes a fusion protein, wherein the fusion protein has a formula of R2-R1 or R2-L-R1, wherein:

R1 is a human fibroblast growth factor 21;

R2 is Exendin-4; and

L is a linker.

8. The host cell according to claim 7, which is selected from E. coli, yeast and CHO.

9. A method for preparing a fusion protein, comprising steps of:

(a) cultivating a host cell selected from E. coli, yeast and CHO to express said fusion protein, wherein the host cell contains a vector having a polynucleotide encoding the fusion protein, wherein the fusion protein has a formula of R2-R1 or R2-L-R1, wherein;

R1 is a human fibroblast growth factor 21;

R2 is Exendin-4; and

L is a linker; and

(b) separating and purifying said fusion protein.

10. A fusion protein having a formula of R2-R1 or R2-L-R1, wherein:

R1 is a mutant of human fibroblast growth factor 21 comprising the amino acid sequence of SEQ ID NO;

1 having up to four N-terminal amino acids deleted from the amino acid sequence of SEQ ID NO;

1, and has the bioactivity and function of FGF-21 comprising the amino acid sequence of SEQ ID NO;

1;

R2 is Exendin-4; and

L is a linker.