Fusion proteins

Patent metadata

Field	Value
Publication number	US-10709766-B2
Application number	US-201715820608-A
Country	US
Kind code	B2
Filing date	Nov 22, 2017
Priority date	May 7, 2015
Publication date	Jul 14, 2020
Grant date	Jul 14, 2020

Abstract

The present invention relates to fusion proteins comprising an insulin receptor agonist fused to a human IgG Fc region through the use of a peptide linker, and the use of such fusion proteins in the treatment of diabetes. The fusion protein of the present invention has an extended time action profile and is useful for providing basal glucose control for an extended period of time.

First claim

We claim:

1. A fusion protein comprising:
a) an insulin receptor agonist having the general formula Z 1 —Z 2 —Z 3 , wherein:
i) Z 1 is an insulin B-chain analog, comprising the amino acid sequence:

X 1 X 2 X 3 QHLCGSHLVEALX 4 LVCGERGFX 5 YX 6 X 7 X 8 X 9

 wherein X 1 is F, Q or A; X 2 is V or G; X 3 is N, K, D, G, Q, A or E; X 4 is E, Y, Q, or H; X 5 is H or F; X 6 is G, T, S, H, V or is absent; X 7 is G, E, P, K, D, S, H or is absent; X 8 is G, E, K, P, Q, D, H or is absent; X 9 is G, T, S, E, K, A or is absent, provided that the insulin B-chain analog includes at least one modification from the amino acid sequence of the B-chain of a molecule of human insulin at X 4 , X 5 , X 6 , X 7 , X 8 , or X 9 (SEQ ID NO:1);
ii) Z 2 is a first peptide linker comprising 5 to 10 amino acids, wherein at least 5 of said amino acids are G residues; and
iii) Z 3 is an insulin A-chain analog comprising the amino acid sequence:

GIVEQCCTSX 1 CSLX 2 QLENYCX 3 X 4

 wherein X 1 is T or I; X 2 is D, Y, Q or E; X 3 is G, N, S or A; and X 4 is any naturally occurring amino acid, or is absent, provided that if X 3 is N, then X 4 must be an amino acid other than G or N (SEQ ID NO:2);
b) a second peptide linker; and
c) a human IgG Fc region;
wherein the C-terminal residue of the insulin receptor agonist is directly fused to the N-terminal residue of the second peptide linker, and the C-terminal residue of the second peptide linker is directly fused to the N-terminal residue of the human IgG Fc region.

2. The fusion protein of claim 1 , wherein:
the insulin B-chain analog includes at least one modification from the amino acid sequence of the human insulin B-chain at X 4 or X 5 of SEQ ID NO:1; and
and the insulin A-chain analog includes at least one modification from the amino acid sequence of the human insulin A-chain at X 1 or X 2 of SEQ ID NO:2.

3. The fusion protein of claim 2 , wherein:
the insulin B-chain analog comprises the sequence of SEQ ID NO:1, wherein: X 1 is F; X 2 is V, X 3 is N or D; X 4 is E; X 5 is H; and
the insulin A-chain analog comprises the sequence of SEQ ID NO:2, wherein: X 1 is I or T; X 2 is D; X 3 is G; and X 4 is absent.

4. The fusion protein of claim 3 , wherein the insulin B-chain analog comprises the sequence of SEQ ID NO:1, wherein X 6 -X 9 are each G.

5. The fusion protein of claim 1 , wherein the first peptide linker comprises the following amino acid sequence:

X 1 GX 2 GGGG

wherein X 1 is G or is absent; and X 2 is G, S or is absent (SEQ ID NO:3).

6. The fusion protein of claim 5 , wherein X 1 and X 2 of SEQ ID NO:3 are G and S, respectively.

7. The fusion protein of claim 1 , wherein the insulin receptor agonist has the following amino acid sequence:

(SEQ ID NO: 5)

FVNQHLCGSHLVEALELVCGERGFHYGGGGGGSGGGGGIVEQCCTSTCSL

DQLENYCG.

8. The fusion protein of claim 1 , wherein the second peptide linker is a peptide having between 10 and 25 amino acids, wherein at least 50% of said amino acids are G residues.

9. The fusion protein of claim 8 , wherein the second peptide linker comprises a peptide having the sequence [GGGGX] n
wherein X is Q, E or S; and wherein n is 2-5 (SEQ ID NO: 26).

10. The fusion protein of claim 9 , wherein the second peptide linker comprises the following amino acid sequence:

GGGGX 1 GGGGX 2 GGGGX 3 GGGGX 4 X 5 X 6

X 1 is Q or E
X 2 is Q or E
X 3 is Q or E
X 4 is G, E, Q or is absent
X 5 is G or absent; and
X 6 is G or is absent
(SEQ ID NO:6).

11. The fusion protein of claim 1 , wherein the second peptide linker has the following amino acid sequence:

(SEQ ID NO: 7)

GGGGQGGGGQGGGGQGGGGG.

12. The fusion protein of claim 1 , wherein the human IgG Fc region is an Fc region from an IgG1, IgG2 or IgG4.

13. The fusion protein of claim 1 , wherein the human IgG Fc region comprises an amino acid sequence selected from the group consisting of SEQ ID NO:8, SEQ ID NO:9 and SEQ ID NO:10.

Assignees

• Lilly Co Eli

Inventors

• Baldwin David Bruce
• Beals John Michael
• Day Jonathan Wesley
• Dickinson Craig Duane
• Korytko Andrew Ihor
• Lazar Gregory Alan

Classifications

• C07K14/62 Primary

  Insulins · CPC title

• C07K2319/30

  Non-immunoglobulin-derived peptide or protein having an immunoglobulin constant or Fc region, or a fragment thereof, attached thereto · CPC title

• A61P5/50

  for increasing or potentiating the activity of insulin · CPC title

• A61K38/28 Primary

  Insulins · CPC title

• A61K38/26

  Glucagons · CPC title