Long-acting conjugate of triple glucagon/GLP-1/GIP receptor agonist

1. A compound of the following Chemical Formula (1):

X—La—F  Chemical Formula (1)

wherein

X is a peptide having activities to a glucagon receptor, a glucagon-like peptide-1 (GLP-1) receptor, and a glucose-dependent insulinotropic polypeptide (GIP) receptor;

L is a polyethylene glycol linker;

a is 1; and

F is an immunoglobulin Fc region;

wherein said F and X are bound to each other through L via a covalent chemical bond; and

wherein the peptide comprises an amino acid sequence of the following Formula (1):

(SEQ ID NO: 103; Formula (1))

Xaa1-Xaa2-Xaa3-Gly-Thr-Phe-Xaa7-Ser-Asp-Xaa10-

Ser-Xaa12-Xaa13-Xaa14-Xaa15-Xaa16-Xaa17-Xaa18-

Xaa19-Xaa20-Xaa21-Phe-Xaa23-Xaa24-Trp-Leu-

Xaa27-Xaa28-Xaa29-Xaa30-R1 

wherein, in Formula (1),

Xaa1 is His, 4-imidazoacetyl (CA), or Tyr;

Xaa2 is Gly, α-methyl-glutamic acid, or aminoisobutyric acid (Aib);

Xaa3 is Glu or Gln;

Xaa7 is Thr or Ile;

Xaa10 is Leu, Tyr, Lys, Cys, or Val;

Xaa12 is Lys, Ser, or Ile;

Xaa13 is Gln, Tyr, Ala, or Cys;

Xaa14 is Leu, Met, or Tyr;

Xaa15 is Cys, Asp, Glu, or Leu;

Xaa16 is Gly, Glu, or Ser;

Xaa17 is Gln, Arg, Ile, Glu, Cys, or Lys;

Xaa18 is Ala, Arg, or His;

Xaa19 is Ala, Gln, Cys, or Val;

Xaa20 is Lys, Gln, or Arg;

Xaa21 is Glu, Gln, Leu, Cys, or Asp;

Xaa23 is Ile or Val;

Xaa24 is Ala, Gln, Cys, Asn, Asp, or Glu;

Xaa27 is Val, Leu, or Lys;

Xaa28 is Cys, Lys, Ala, Asn, or Asp;

Xaa29 is Cys, Gly, Gln, Thr, Glu, or His;

Xaa30 is Cys, Gly, Lys, or His, or is absent; and

R1 is Cys, GKKNDWKHNIT (SEQ ID NO: 106), m-SSGAPPPS-n (SEQ ID NO: 107), or m-SSGQPPPS-n (SEQ ID NO: 108), or is absent;

wherein

m is -Cys-, -Pro-, or -Gly-Pro-; and

n is -Cys-, -Gly-, -Ser-, or -His-Gly-, or is absent, and

wherein an in vitro activity of 1% or more is exhibited by the peptide X, in its isolated free from, at the glucagon receptor compared to that of native glucagon, and/or at the GIP receptor compared to that of native GIP, and/or at the GLP-1 receptor compared to that of native GLP-1.

2. The compound of claim 1 , wherein R1 is GKKNDWKHNIT (SEQ ID NO: 106).

3. The compound of claim 1 , wherein, in Formula (1),

Xaa1 is 4-imidazoacetyl, histidine, or tyrosine;

Xaa2 is glycine, α-methyl-glutamic acid, or Aib;

Xaa3 is Glu or Gln;

Xaa7 is Thr or Ile;

Xaa10 is Tyr or Cys;

Xaa13 is Ala, Gln, Tyr, or Cys;

Xaa14 is Leu, Met, or Tyr;

Xaa15 is Asp, Glu, or Leu;

Xaa16 is Gly, Glu, or Ser;

Xaa17 is Gln, Arg, Ile, Glu, Cys, or Lys;

Xaa18 is Ala, Gln, Arg, or His;

Xaa19 is Ala, Gln, Cys, or Val;

Xaa20 is Lys, Gln, or Arg;

Xaa21 is Cys, Glu, Gln, Leu, or Asp;

Xaa23 is Ile or Val;

Xaa24 is Cys, Ala, Gln, Asp, or Glu;

Xaa27 is Leu;

Xaa28 is Lys, Cys, Asp, or Asp;

Xaa29 is Gly, Gln, CYs, or His;

Xaa30 is Cys, Gly, Lys, or His;

Xaa31 is Pro or Cys; and

Xaa40 is Cys or is absent.

4. The compound of claim 3 , wherein, in Formula (1),

Xaa2 is α-methyl-glutamic acid or Aib;

Xaa3 is Gln;

Xaa7 is Thr;

Xaa10 is Tyr;

Xaa12 is Lys;

Xaa13 is Tyr, Ala, or Cys;

Xaa14 is Leu or Met;

Xaa15 is Cys, Asp, or Glu;

Xaa16 is Gly or Glu;

Xaa19 is Ala, Gln, or Cys;

Xaa20 is Lys or Gln;

Xaa21 is Glu, Gln, Cys, or Asp;

Xaa24 is Ala, Gln, Cys, or Asn; and

Xaa27 is Leu or Lys.

5. The compound of claim 1 , wherein X is a peptide comprising an amino acid sequence selected from the group consisting of SEQ ID NOS: 1 to 11, 14 to 16, 21 to 86, 88-91, 93, and 95 to 102.

6. The compound of claim 1 , wherein X is a peptide comprising an amino acid sequence selected from the group consisting of SEQ ID NOS: 21, 22, 23, 31, 32, 37, 42, 43, 50, 53, 55, 64 to 77, 79, and 96 to 102.

7. The compound of claim 3 ,

wherein, in Formula (1),

Xaa13 is Ala, Tyr, or Cys;

Xaa15 is Asp or Glu;

Xaa17 is Gln, Arg, Cys, or Lys;

Xaa18 is Ala, Arg, or His;

Xaa21 is Cys, Glu, or Asp;

Xaa24 is Cys, Gln, or Asn;

Xaa28 is Cys, Asn, or Asp;

Xaa29 is Gln, Cys, or His; and

Xaa30 is Cys, Lys, or His.

8. The compound of claim 3 ,

wherein, in Formula (1),

Xaa1 is His or 4-imidazoacetyl;

Xaa13 is Ala or Cys;

Xaa14 is Met;

Xaa15 is Asp;

Xaa16 is Glu;

Xaa17 is Ile or Lys;

Xaa18 is Ala or His;

Xaa19 is Gln or Cys;

Xaa20 is Lys;

Xaa21 is Asp;

Xaa23 is Val;

Xaa24 is Asn;

Xaa28 is Ala or Asn;

Xaa29 is Gln or Thr; and

Xaa30 is Cys or Lys, or is absent.

9. The compound according to claim 1 , wherein F is an immunoglobulin Fc region in a dimeric form and one molecule of X is covalently linked to one Fc region in the dimeric form.

10. A pharmaceutical composition comprising the compound of claim 1 .

11. A method of treating a metabolic syndrome comprising administering the compound according to claim 1 to a subject in need thereof.

12. The method of claim 11 , wherein the metabolic syndrome comprises impaired glucose tolerance, hypercholesterolemia, dyslipidemia, obesity, diabetes, hypertension, arteriosclerosis due to dyslipidemia, atherosclerosis, arteriosclerosis, or coronary heart disease.

13. A compound of the following Chemical Formula (1):

X—La—F  Chemical Formula (1)

wherein

X is a peptide having activities to a glucagon receptor, a glucagon-like peptide-1 (GLP-1) receptor, and a glucose-dependent insulinotropic polypeptide (GIP) receptor;

L is a linker;

a is 0 or a positive integer, with the proviso that when a is 2 or greater, each L is independent from each other; and

F is a material capable of increasing the half-life of the peptide;

wherein the peptide comprises an amino acid sequence having sequence identity of 90% or higher to the sequence of the following Formula (3):

(SEQ ID NO: 105; Formula (3))

Xaa1-Xaa2-Gln-Gly-Thr-Phe-Thr-Ser-Asp-Tyr-Ser-

Lys-Xaa13-Leu-Asp-Glu-Xaa17-Xaa18-Xaa19-Lys-

Xaa21-Phe-Val-Xaa24-Trp-Leu-Leu-Xaa28-Xaa29-

Xaa30-Xaa31-Ser-Ser-Gly-Gln-Pro-Pro-Pro-Ser-

Xaa40

wherein, in Formula (3),

Xaa1 is His or Tyr;

Xaa2 is α-methyl-glutamic acid or aminoisobutyric acid (Aib);

Xaa13 is Ala, Tyr, or Cys;

Xaa17 is Arg, Cys, or Lys;

Xaa18 is Ala or Arg;

Xaa19 is Ala or Cys;

Xaa21 is Glu or Asp;

Xaa24 is Gln or Asn;

Xaa28 is Cys or Asp;

Xaa29 is Cys, His, or Gln;

Xaa30 is Cys or His;

Xaa31 is Pro or Cys; and

Xaa40 is Cys or is absent.

14. The compound of claim 13 , wherein X is a peptide comprising an amino acid sequence selected from the group consisting of SEQ ID NOS: 21, 22, 42, 43, 50, 64, 66, 67, 70, 71, 76, 77, 96, 97, and 100.

15. The compound according to claim 13 , wherein, in Formula 3, the 16 th amino acid and the 20 th amino acid from the N-terminus together form a ring.

16. The compound of claim 13 , wherein Xaa1 is Tyr.

17. A compound of the following Chemical Formula (1):

X—La—F  Chemical Formula (1)

wherein

X is a peptide having activities to a glucagon receptor, a glucagon-like peptide-1 (GLP-1) receptor, and a glucose-dependent insulinotropic polypeptide (GIP) receptor;

L is a linker;

a is 0 or a positive integer, with the proviso that when a is 2 or greater, each L is independent from each other; and

F is a material capable of increasing the half-life of the peptide;

wherein the peptide comprises an amino acid sequence having sequence identity of 95% or greater to the sequence of the following Formula (1):

(SEQ ID NO: 103; Formula (1))

Xaa1-Xaa2-Xaa3-Gly-Thr-Phe-Xaa7-Ser-Asp-Xaa10-

Ser-Xaa12-Xaa13-Xaa14-Xaa15-Xaa16-Xaa17-Xaa18-

Xaa19-Xaa20-Xaa21-Phe-Xaa23-Xaa24-Trp-Leu-

Xaa27-Xaa28-Xaa29-Xaa30-R1

wherein, in Formula (1),

Xaa1 is His, 4-imidazoacetyl (CA), or Tyr;

Xaa2 is α-methyl-glutamic acid, or aminoisobutyric acid (Aib);

Xaa3 is Gln;

Xaa7 is Thr;

Xaa10 is Tyr;

Xaa12 is Lys;

Xaa13 is Tyr, Ala, or Cys;

Xaa14 is Leu or Met;

Xaa15 is Cys, Asp, or Glu;

Xaa16 is Gly or Glu;

Xaa17 is Gln, Arg, Ile, Glu, Cys, or Lys;

Xaa18 is Ala, Arg, or His;

Xaa19 is Ala, Gln, or Cys;

Xaa20 is Lys or Gln;

Xaa21 is Glu, Gln, Cys, or Asp;

Xaa23 is Ile or Val;

Xaa24 is Ala, Gln, Cys, or Asn;

Xaa27 is Leu or Lys;

Xaa28 is Cys, Lys, Ala, Asn, or Asp;

Xaa29 is Cys, Gly, Gln, Thr, Glu, or His;

Xaa30 is Cys, Gly, Lys, or His, or is absent; and

R1 is Cys, GKKNDWKHNIT (SEQ ID NO: 106), m-SSGAPPPS-n (SEQ ID NO: 107), or m-SSGQPPPS-n (SEQ ID NO: 108), or is absent;

wherein

m is -Cys-, -Pro-, or -Gly-Pro-; and

n is -Cys-, -Gly-, -Ser-, or -His-Gly-, or is absent.

18. The compound of claim 17 , wherein R1 is GKKNDWKHNIT (SEQ ID NO: 106).

19. The compound of claim 17 , wherein X is a peptide comprising an amino acid sequence selected from the group consisting of SEQ ID NOS: 21 to 24, 27 to 32, 34, 36, 37, 39, 42, 43, 50 to 52, 56, 58, 64 to 71, 73 to 78, 81, 82, 86, 88, 89, 93, and 95 to 102.

20. The compound of claim 17 , wherein X is a peptide comprising an amino acid sequence selected from the group consisting of SEQ ID NOS: 21, 22, 23, 31, 32, 42, 43, 50, 53, 55, 64 to 77, 79, and 96 to 102.

21. The compound of claim 17 ,

wherein, in Formula (1),

Xaa13 is Ala, Tyr, or Cys;

Xaa15 is Asp or Glu;

Xaa17 is Gln, Arg, Cys, or Lys;

Xaa18 is Ala, Arg, or His;

Xaa21 is Cys, Glu, or Asp;

Xaa23 is Ile or Val;

Xaa24 is Cys, Gln, or Asn;

Xaa28 is Cys, Asn, or Asp;

Xaa29 is Gln, Cys, or His; and

Xaa30 is Cys, Lys, or His.

22. The compound of claim 17 ,

wherein, in Formula (1),

Xaa1 is His or CA;

Xaa13 is Ala or Cys;

Xaa14 is Met;

Xaa15 is Asp;

Xaa16 is Glu;

Xaa17 is Ile or Lys;

Xaa18 is Ala or His;

Xaa19 is Gln or Cys;

Xaa20 is Lys;

Xaa21 is Asp;

Xaa23 is Val;

Xaa24 is Asn;

Xaa28 is Ala or Asn;

Xaa29 is Gln or Thr; and

Xaa30 is Cys or Lys, or is absent.

23. The compound according to claim 13 , wherein F is selected from the group consisting of a polymer, fatty acid, cholesterol, albumin and a fragment thereof, an albumin-binding material, a polymer of repeating units of particular amino acid sequences, an antibody, an antibody fragment, an FcRn-binding material, an in vivo connective tissue, a nucleotide, fibronectin, transferrin, a saccharide, heparin, and elastin.

24. The compound according to claim 23 , wherein F is an immunoglobulin Fc region.

25. The compound according to claim 13 , wherein L is a peptide, fatty acid, a saccharide, a polymer, a low molecular weight compound, a nucleotide, or a combination thereof.

26. The compound of claim 25 , wherein the polymer is selected from the group consisting of polyethylene glycol, polypropylene glycol, an ethylene glycol-propylene glycol copolymer, polyoxyethylated polyol, polyvinyl alcohol, polysaccharide, dextran, polyvinyl ethyl ether, a biodegradable polymer, a lipid polymer, chitins, hyaluronic acid, an oligonucleotide, and a combination thereof.

27. A pharmaceutical composition comprising the compound of claim 13 .

28. A method of treating metabolic syndrome comprising administering the compound according to claim 14 to a subject in need thereof.

29. The method of claim 28 , wherein the metabolic syndrome comprises impaired glucose tolerance, hypercholesterolemia, dyslipidemia, obesity, diabetes, hypertension, arteriosclerosis due to dyslipidemia, atherosclerosis, arteriosclerosis, or coronary heart disease.

30. The compound according to claim 17 , wherein F is selected from the group consisting of a polymer, fatty acid, cholesterol, albumin and a fragment thereof, an albumin-binding material, a polymer of repeating units of particular amino acid sequences, an antibody, an antibody fragment, an FcRn-binding material, an in vivo connective tissue, a nucleotide, fibronectin, transferrin, a saccharide, heparin, and elastin.

31. The compound according to claim 30 , wherein F is an immunoglobulin Fc region.

32. The compound according to claim 17 , wherein L is a peptide, fatty acid, a saccharide, a polymer, a low molecular weight compound, a nucleotide, or a combination thereof.

33. The compound of claim 32 , wherein the polymer is selected from the group consisting of polyethylene glycol, polypropylene glycol, an ethylene glycol-propylene glycol copolymer, polyoxyethylated polyol, polyvinyl alcohol, polysaccharide, dextran, polyvinyl ethyl ether, a biodegradable polymer, a lipid polymer, chitins, hyaluronic acid, an oligonucleotide, and a combination thereof.

34. A pharmaceutical composition comprising the compound of claim 17 .

35. A method of treating metabolic syndrome comprising administering the compound according to claim 17 to a subject in need thereof.

36. The method of claim 35 , wherein the metabolic syndrome comprises impaired glucose tolerance, hypercholesterolemia, dyslipidemia, obesity, diabetes, hypertension, arteriosclerosis due to dyslipidemia, atherosclerosis, arteriosclerosis, or coronary heart disease.

37. A compound of the following Chemical Formula (1):

X—La—F  Chemical Formula (1)

wherein

X is a peptide selected from the peptides of the amino acid sequence of SEQ ID NOS: 1-102;

L is a linker;

a is 0 or a positive integer, with the proviso that when a is 2 or greater, each L is independent from each other; and

F is a material capable of increasing the half-life of the peptide.

38. The compound of claim 1 , wherein the peptide, in its isolated free form, exhibits an in vitro activity of 1% or higher compared to in vitro activity of native glucagon at the glucagon receptor and in vitro activity of native GLP-1 at the GLP-1 receptor, respectively.

39. The compound of claim 1 , wherein the peptide, in its isolated free form, exhibits an in vitro activity of 1% or higher compared to in vitro activity of native glucagon at the glucagon receptor and in vitro activity of native GIP at the GIP-receptor, respectively.

40. The compound of claim 1 , wherein the peptide, in its isolated free form, exhibits an in vitro activity of 1% or higher compared to in vitro activity of native GLP-1 at the GLP-1 receptor and in vitro activity of native GIP at the GIP-receptor, respectively.

41. The compound of claim 1 , wherein the peptide, in its isolated free form, exhibits an in vitro activity of 1% or higher compared to in vitro activity of native glucagon at the glucagon receptor, in vitro activity of native GLP-1 at the GLP-1 receptor, and in vitro activity of native GIP at the GIP-receptor, respectively.