IP Library › Granted Patent US 8,273,854
Granted Patent B2
US 8,273,854 · App. 12/262,832 · Granted Sep 25, 2012

GLP-1 analog fusion proteins

Assignee: Eli Lilly and Company
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Quick Facts
Patent No.
US 8,273,854
App. No.
12/262,832
Granted
Sep 25, 2012
Kind
B2
Abstract

The invention provides specific GLP-1 analogs fused to specific IgG4-Fc derivatives. These fusion proteins have an increased half-life, decreased immunogenicity, and reduce effector activity. The fusion proteins are useful in treating diabetes, obesity, irritable bowel syndrome and other conditions that would be benefited by lowering plasma glucose, inhibiting gastric and/or intestinal motility and inhibiting gastric and/or intestinal emptying, or inhibiting food intake.

Claims (147)

1. A method of treating a patient with non-insulin dependent diabetes mellitus comprising administering to the patient a therapeutically effective amount of a heterologous fusion protein comprising:

a GLP-1 analog comprising SEQ ID NO: 1

His-Xaa 8 -Glu-Gly-Thr-Phe-Thr-Ser-Asp-Val-Ser-Ser-

Tyr-Leu-Glu-Glu-Gln-Ala-Ala-Lys-Glu-Phe-Ile-Ala-

Trp-Leu-Val-Lys-Gly-Gly-Gly

wherein Xaa 8 is Gly;

an Fc portion of an immunoglobulin comprising SEQ ID NO: 7

Ala-Glu-Ser-Lys-Tyr-Gly-Pro-Pro-Cys-Pro-Pro-Cys-

Pro-Ala-Pro-Xaa 16 -Xaa 17 -Xaa 18 -Gly-Gly-Pro-Ser-Val-

Phe-Leu-Phe-Pro-Pro-Lys-Pro-Lys-Asp-Thr-Leu-Met-

Ile-Ser-Arg-Thr-Pro-Glu-Val-Thr-Cys-Val-Val-Val-

Asp-Val-Ser-Gln-Glu-Asp-Pro-Glu-Val-Gln-Phe-Asn-

Trp-Tyr-Val-Asp-Gly-Val-Glu-Val-His-Asn-Ala-Lys-

Thr-Lys-Pro-Arg-Glu-Glu-Gln-Phe-Xaa 80 -Ser-Thr-Tyr-

Arg-Val-Val-Ser-Val-Leu-Thr-Val-Leu-His-Gln-Asp-

Trp-Leu-Asn-Gly-Lys-Glu-Tyr-Lys-Cys-Lys-Val-Ser-

Asn-Lys-Gly-Leu-Pro-Ser-Ser-Ile-Glu-Lys-Thr-Ile-

Ser-Lys-Ala-Lys-Gly-Gln-Pro-Arg-Glu-Pro-Gln-Val-

Tyr-Thr-Leu-Pro-Pro-Ser-Gln-Glu-Glu-Met-Thr-Lys-

Asn-Gln-Val-Ser-Leu-Thr-Cys-Leu-Val-Lys-Gly-Phe-

Tyr-Pro-Ser-Asp-Ile-Ala-Val-Glu-Trp-Glu-Ser-Asn-

Gly-Gln-Pro-Glu-Asn-Asn-Tyr-Lys-Thr-Thr-Pro-Pro-

Val-Leu-Asp-Ser-Asp-Gly-Ser-Phe-Phe-Leu-Tyr-Ser-

Arg-Leu-Thr-Val-Asp-Lys-Ser-Arg-Trp-Gln-Glu-Gly-

Asn-Val-Phe-Ser-Cys-Ser-Val-Met-His-Glu-Ala-Leu-

His-Asn-His-Tyr-Thr-Gln-Lys-Ser-Leu-Ser-Leu-Ser-

Leu-Gly-Xaa 230

wherein:

Xaa at position 16 is Glu;

Xaa at position 17 is Ala;

Xaa at position 18 is Ala;

Xaa at position 80 is Asn; and

Xaa at position 230 is Lys or absent;

and a peptide linker comprising SEQ ID NO: 8

Gly-Gly-Gly-Gly-Ser-Gly-Gly-Gly-Gly-Ser-Gly-Gly-

Gly-Gly-Ser

wherein the N-terminal glycine of the peptide linker is directly fused to the C-terminal glycine residue of the GLP-1 analog and the C-terminal serine of the peptide linker is directly fused to N-terminal alanine of the Fc portion.

2. A method of treating a patient with non-insulin dependent diabetes mellitus comprising administering to the patient a therapeutically effective amount of a heterologous fusion protein consisting of:

a GLP-1 analog consisting of SEQ ID NO:1

His-Xaa 8 -Glu-Gly-Thr-Phe-Thr-Ser-Asp-Val-Ser-Ser-

Tyr-Leu-Glu-Glu-Gln-Ala-Ala-Lys-Glu-Phe-Ile-Ala-

Trp-Leu-Val-Lys-Gly-Gly-Gly

wherein Xaa 8 is Gly;

an Fc portion of an immunoglobulin consisting of SEQ ID NO:7

Ala-Glu-Ser-Lys-Tyr-Gly-Pro-Pro-Cys-Pro-Pro-Cys-

Pro-Ala-Pro-Xaa 16 -Xaa 17 -Xaa 18 -Gly-Gly-Pro-Ser-Val-

Phe-Leu-Phe-Pro-Pro-Lys-Pro-Lys-Asp-Thr-Leu-Met-

Ile-Ser-Arg-Thr-Pro-Glu-Val-Thr-Cys-Val-Val-Val-

Asp-Val-Ser-Gln-Glu-Asp-Pro-Glu-Val-Gln-Phe-Asn-

Trp-Tyr-Val-Asp-Gly-Val-Glu-Val-His-Asn-Ala-Lys-

Thr-Lys-Pro-Arg-Glu-Glu-Gln-Phe-Xaa 80 -Ser-Thr-Tyr-

Arg-Val-Val-Ser-Val-Leu-Thr-Val-Leu-His-Gln-Asp-

Trp-Leu-Asn-Gly-Lys-Glu-Tyr-Lys-Cys-Lys-Val-Ser-

Asn-Lys-Gly-Leu-Pro-Ser-Ser-Ile-Glu-Lys-Thr-Ile-

Ser-Lys-Ala-Lys-Gly-Gln-Pro-Arg-Glu-Pro-Gln-Val-

Tyr-Thr-Leu-Pro-Pro-Ser-Gln-Glu-Glu-Met-Thr-Lys-

Asn-Gln-Val-Ser-Leu-Thr-Cys-Leu-Val-Lys-Gly-Phe-

Tyr-Pro-Ser-Asp-Ile-Ala-Val-Glu-Trp-Glu-Ser-Asn-

Gly-Gln-Pro-Glu-Asn-Asn-Tyr-Lys-Thr-Thr-Pro-Pro-

Val-Leu-Asp-Ser-Asp-Gly-Ser-Phe-Phe-Leu-Tyr-Ser-

Arg-Leu-Thr-Val-Asp-Lys-Ser-Arg-Trp-Gln-Glu-Gly-

Asn-Val-Phe-Ser-Cys-Ser-Val-Met-His-Glu-Ala-Leu-

His-Asn-His-Tyr-Thr-Gln-Lys-Ser-Leu-Ser-Leu-Ser-

Leu-Gly-Xaa 230

wherein:

Xaa at position 16 is Glu;

Xaa at position 17 is Ala;

Xaa at position 18 is Ala;

Xaa at position 80 is Asn; and

Xaa at position 230 is Lys or absent;

and a peptide linker consisting of SEQ ID NO:8

Gly-Gly-Gly-Gly-Ser-Gly-Gly-Gly-Gly-Ser-Gly-Gly-

Gly-Gly-Ser

wherein the N-terminal glycine of the peptide linker is directly fused to the C-terminal glycine residue of the GLP-1 analog and the C-terminal serine of the peptide linker is directly fused to N-terminal alanine of the Fc portion.

3. A method of treating a patient with non-insulin dependent diabetes mellitus comprising administering to the patient a therapeutically effective amount of a heterologous fusion protein comprising a dimer comprising two chains joined together by disulfide bonds wherein each chain has:

a GLP-1 analog consisting of SEQ ID NO: 1

His-Xaa 8 -Glu-Gly-Thr-Phe-Thr-Ser-Asp-Val-Ser-Ser-

Tyr-Leu-Glu-Glu-Gln-Ala-Ala-Lys-Glu-Phe-Ile-Ala-

Trp-Leu-Val-Lys-Gly-Gly-Gly,

wherein Xaa 8 is Gly;

an Fc portion of an immunoglobulin comprising SEQ ID NO:7

Ala-Glu-Ser-Lys-Tyr-Gly-Pro-Pro-Cys-Pro-Pro-Cys-

Pro-Ala-Pro-Xaa 16 -Xaa 17 -Xaa 18 -Gly-Gly-Pro-Ser-Val-

Phe-Leu-Phe-Pro-Pro-Lys-Pro-Lys-Asp-Thr-Leu-Met-

Ile-Ser-Arg-Thr-Pro-Glu-Val-Thr-Cys-Val-Val-Val-

Asp-Val-Ser-Gln-Glu-Asp-Pro-Glu-Val-Gln-Phe-Asn-

Trp-Tyr-Val-Asp-Gly-Val-Glu-Val-His-Asn-Ala-Lys-

Thr-Lys-Pro-Arg-Glu-Glu-Gln-Phe-Xaa 80 -Ser-Thr-Tyr-

Arg-Val-Val-Ser-Val-Leu-Thr-Val-Leu-His-Gln-Asp-

Trp-Leu-Asn-Gly-Lys-Glu-Tyr-Lys-Cys-Lys-Val-Ser-

Asn-Lys-Gly-Leu-Pro-Ser-Ser-Ile-Glu-Lys-Thr-Ile-

Ser-Lys-Ala-Lys-Gly-Gln-Pro-Arg-Glu-Pro-Gln-Val-

Tyr-Thr-Leu-Pro-Pro-Ser-Gln-Glu-Glu-Met-Thr-Lys-

Asn-Gln-Val-Ser-Leu-Thr-Cys-Leu-Val-Lys-Gly-Phe-

Tyr-Pro-Ser-Asp-Ile-Ala-Val-Glu-Trp-Glu-Ser-Asn-

Gly-Gln-Pro-Glu-Asn-Asn-Tyr-Lys-Thr-Thr-Pro-Pro-

Val-Leu-Asp-Ser-Asp-Gly-Ser-Phe-Phe-Leu-Tyr-Ser-

Arg-Leu-Thr-Val-Asp-Lys-Ser-Arg-Trp-Gln-Glu-Gly-

Asn-Val-Phe-Ser-Cys-Ser-Val-Met-His-Glu-Ala-Leu-

His-Asn-His-Tyr-Thr-Gln-Lys-Ser-Leu-Ser-Leu-Ser-

Leu-Gly-Xaa 230 ,

wherein Xaa at position 16 is Glu;

Xaa at position 17 is Ala;

Xaa at position 18 is Ala;

Xaa at position 80 is Asn; and

Xaa at position 230 is absent; and

a peptide linker consisting of SEQ ID NO: 8

Gly-Gly-Gly-Gly-Ser-Gly-Gly-Gly-Gly-Ser-Gly-Gly-

Gly-Gly-Ser

wherein the N-terminal glycine of the peptide linker is directly fused to the C-terminal glycine residue of the GLP-1 analog and the C-terminal serine of the peptide linker is directly fused to N-terminal alanine of the Fc portion.

4. A method of treating a patient with non-insulin dependent diabetes mellitus comprising administering to the patient a therapeutically effective amount of a heterologous fusion protein comprising a dimer comprising two chains joined together by disulfide bonds wherein each chain has:

a GLP-1 analog consisting of SEQ ID NO:1

His-Xaa 8 -Glu-Gly-Thr-Phe-Thr-Ser-Asp-Val-Ser-Ser-

Tyr-Leu-Glu-Glu-Gln-Ala-Ala-Lys-Glu-Phe-Ile-Ala-

Trp-Leu-Val-Lys-Gly-Gly-Gly,

wherein Xaa 8 is Gly;

Ala-Glu-Ser-Lys-Tyr-Gly-Pro-Pro-Cys-Pro-Pro-Cys-

Pro-Ala-Pro-Xaa 16 -Xaa 17 -Xaa 18 -Gly-Gly-Pro-Ser-Val-

Phe-Leu-Phe-Pro-Pro-Lys-Pro-Lys-Asp-Thr-Leu-Met-

Ile-Ser-Arg-Thr-Pro-Glu-Val-Thr-Cys-Val-Val-Val-

Asp-Val-Ser-Gln-Glu-Asp-Pro-Glu-Val-Gln-Phe-Asn-

Trp-Tyr-Val-Asp-Gly-Val-Glu-Val-His-Asn-Ala-Lys-

Thr-Lys-Pro-Arg-Glu-Glu-Gln-Phe-Xaa 80 -Ser-Thr-Tyr-

Arg-Val-Val-Ser-Val-Leu-Thr-Val-Leu-His-Gln-Asp-

Trp-Leu-Asn-Gly-Lys-Glu-Tyr-Lys-Cys-Lys-Val-Ser-

Asn-Lys-Gly-Leu-Pro-Ser-Ser-Ile-Glu-Lys-Thr-Ile-

Ser-Lys-Ala-Lys-Gly-Gln-Pro-Arg-Glu-Pro-Gln-Val-

Tyr-Thr-Leu-Pro-Pro-Ser-Gln-Glu-Glu-Met-Thr-Lys-

Asn-Gln-Val-Ser-Leu-Thr-Cys-Leu-Val-Lys-Gly-Phe-

Tyr-Pro-Ser-Asp-Ile-Ala-Val-Glu-Trp-Glu-Ser-Asn-

Gly-Gln-Pro-Glu-Asn-Asn-Tyr-Lys-Thr-Thr-Pro-Pro-

Val-Leu-Asp-Ser-Asp-Gly-Ser-Phe-Phe-Leu-Tyr-Ser-

Arg-Leu-Thr-Val-Asp-Lys-Ser-Arg-Trp-Gln-Glu-Gly-

Asn-Val-Phe-Ser-Cys-Ser-Val-Met-His-Glu-Ala-Leu-

His-Asn-His-Tyr-Thr-Gln-Lys-Ser-Leu-Ser-Leu-Ser-

Leu-Gly-Xaa 230 ,

wherein Xaa at position 16 is Glu;

Xaa at position 17 is Ala;

Xaa at position 18 is Ala;

Xaa at position 80 is Asn; and

Xaa at position 230 is Lys; and

a peptide linker consisting of SEQ ID NO:8

Gly-Gly-Gly-Gly-Ser-Gly-Gly-Gly-Gly-Ser-Gly-Gly-

Gly-Gly-Ser

wherein the N-terminal glycine of the peptide linker is directly fused to the C-terminal glycine residue of the GLP-1 analog and the C-terminal serine of the peptide linker is directly fused to N-terminal alanine of the Fc portion.

5. The method according to any one of claims 1 to 4 wherein the fusion protein is produced by expressing the fusion protein in a mammalian host cell.

6. The method according to claim 5 wherein the host cell is a CHO cell.

Continuity (3)
Division 10558627
Provisional Application 60477880 · Jun 12, 2003
Related Publication 20090074769A1 · Mar 19, 2009