Long-acting co-agonists of the glucagon and GLP-1 receptors

Abstract

Long-acting co-agonists of the glucagon and GLP-1 receptors are described.

Claims

1. A GCG/GLP-1 receptor co-agonist peptide comprising the formula TABLE-US-00017 (SEQ ID NO: 123) HX.sup.2QGTFTSX.sup.9X.sup.10SLYLDX.sup.16RAAX.sup.20X.sup.21FVX.sup.24X.sup.25LX.sup.27X.sup.28TX.sup.30-NH.sub.2 or a pharmaceutically acceptable salt or counterion thereof, wherein X.sup.2 is alpha-aminoisobutyric acid (aib), D-Ser, or alpha-Methyl-L-Serine (alpha-MS); X.sup.9 is Asp, or Glu; X.sup.10 is Lys or p-aminomethyl-L-phenylalanine (pAF) conjugated to a fatty acid provided that the amino acid at position 20 or 24 is a Lys conjugated to a fatty diacid, or Tyr; X.sup.16 is aib, Ala, Ser, or Glu; X.sup.20 is Lys is conjugated to a fatty diacid, pAF conjugated to a fatty diacid, norleucine (Nle) conjugated to a fatty diacid, or Gln; X.sup.21 is Lys conjugated to a fatty diacid, pAF conjugated to a fatty diacid, Asp, alpha-methyl-L-phenylalanine (alpha-MF), or alpha-MD; X.sup.24 is Gln, Lys conjugated to a fatty diacid, or pAF conjugated to a fatty diacid; X.sup.25 is Trp or alpha-methyl-L-tryptophan (alpha-MW); X.sup.27 is L-Met sulphone or Leu; X.sup.28 is Glu, Asp, alpha-MD, Lys, aib, Ala, Lys conjugated to a fatty diacid or pAF conjugated to a fatty diacid; X.sup.30 is absent, or Lys linked at the C-terminus to gamma-Glu when X.sup.27 is Leu or L-Met-sulphone and X.sup.28 is Ala, aib, alpha-MD, or Lys conjugated to a fatty diacid; with the proviso that for each co-agonist peptide only one of X.sup.10, X.sup.20, X.sup.21, X.sup.24, or X.sup.28 is conjugated to a fatty diacid; and wherein the GCG/GLP-1 receptor co-agonist peptide comprises at: 1) X.sup.10 a Lys conjugated to a C16 fatty acid and a Lys at position 20 or 24 conjugated to a fatty diacid; 2) X.sup.20 a pAF conjugated to a fatty diacid or a Lys conjugated to a fatty diacid; 3) X.sup.21 a pAF conjugated to a fatty diacid or a Lys conjugated to a fatty diacid; 4) X.sup.24 a pAF conjugated to a fatty diacid or a Lys conjugated to a fatty diacid; or 5) X.sup.28 a pAF conjugated to a fatty diacid or a Lys conjugated to a fatty diacid.

2. The GCG/GLP-1 receptor co-agonist peptide of claim 1, wherein the fatty diacid comprises a C14, C15, C16, C17, C18, C19, or C20 fatty diacid.

3. The GCG/GLP-1 receptor co-agonist peptide of claim 1, wherein the GCG/GLP-1 receptor co-agonist peptide comprises the fatty diacid conjugated to Lys or pAF via a gamma-Glu linker.

4. The GCG/GLP-1 receptor co-agonist peptide of claim 1, wherein the GCG/GLP-1 receptor co-agonist peptide comprises the fatty diacid conjugated to Lys or pAF via a PEG.sub.2PEG.sub.2-gamma-Glu linker wherein PEG.sub.2 is 8-amino-3,6-dioxaoctanoic acid.

5. The GCG/GLP-1 receptor co-agonist peptide of claim 1, wherein the GCG/GLP-1 receptor co-agonist peptide comprises at X.sup.10 a pAF conjugated to a fatty diacid.

6. The GCG/GLP-1 receptor co-agonist peptide of claim 1, wherein the GCG/GLP-1 receptor co-agonist peptide has activity at the glucagon receptor and the GLP-1 receptor.

7. A composition comprising one or more of the GCG/GLP-1 receptor co-agonist peptides of claim 1, or pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

8. A method for treating a patient for a metabolic disease or disorder comprising administering to a patient in need thereof an effective amount of one or more of the GCG/GLP-1 receptor co-agonist peptides of claim 1 to treat the metabolic disease or disorder in the patient, wherein the metabolic disease or disorder comprises diabetes, non-alcoholic fatty liver disease (NAFLD), non-alcoholic steatohepatitis (NASH), or obesity.

9. The method of claim 8, wherein the diabetes comprises Type I diabetes, Type II diabetes, or gestational diabetes.

10. The method of claim 8, wherein the patient has more than one metabolic disease or disorder.

11. The method of claim 8, wherein the metabolic disease or disorder comprises (i) diabetes and NASH, NAFLD, or obesity; (ii) obesity and NASH or NAFLD; (iii) diabetes, NASH, and obesity; (iv) diabetes, NAFLD, and obesity; or (v) diabetes and obesity.

12. A method for treating a patient for a metabolic disease or disorder comprising administering to the patient in need thereof an effective amount of the composition of claim 7 to treat the metabolic disease or disorder in the patient, wherein the metabolic disease or disorder comprises diabetes, non-alcoholic fatty liver disease (NAFLD), non-alcoholic steatohepatitis (NASH), or obesity.

13. The method of claim 12, wherein the patient has more than one metabolic disease or disorder.

14. The method of claim 12, wherein the metabolic disease or disorder comprises (i) diabetes and NASH, NAFLD, or obesity; (ii) obesity and NASH or NAFLD; (iii) diabetes, NASH, and obesity; (iv) diabetes, NAFLD, and obesity; or (v) diabetes and obesity.

15. A method for treating a metabolic disease or disorder in a patient or individual comprising administering to the patient or individual in need thereof an effective amount of a composition comprising a co-agonist peptide agonist of claim 1 and administering to the patient or individual an effective amount of a composition comprising an insulin or insulin analog to treat the metabolic disease or disorder in the patient or individual wherein the metabolic disease or disorder comprises diabetes, non-alcoholic fatty liver disease (NAFLD), non-alcoholic steatohepatitis (NASH), or obesity.

16. The method of claim 15, wherein the insulin analog comprises insulin detemir, insulin glargine, insulin glulisine, insulin degludec, or insulin lispro.

17. The method of claim 15, wherein the diabetes comprises Type I diabetes, Type II diabetes, or gestational diabetes.

18. The method of claim 15, wherein the patient has more than one metabolic disease or disorder selected from (i) diabetes and NASH, NAFLD, or obesity; (ii) obesity and NASH or NAFLD; (iii) diabetes, NASH, and obesity; (iv) diabetes, NAFLD, and obesity; or (v) diabetes and obesity.

19. A GCG/GLP-1 receptor co-agonist peptide comprising the formula TABLE-US-00018 (SEQ ID NO: 105) HX.sup.2QGTFTSDX.sup.10SLYLDX.sup.16RAAX.sup.20X.sup.21FVX.sup.24X.sup.25LX.sup.27X.sup.28TX.sup.30-NH.sub.2 wherein X.sup.2 is alpha-aminoisobutyric acid (aib), D-Ser, or alpha-Methyl-L-Serine (alpha-MS); X.sup.9 is Asp or alpha-Methyl-L-Aspartic acid (alpha-MD); X.sup.10 is Lys or p-aminomethyl-L-phenylalanine (pAF) conjugated to a fatty acid provided that the amino acid at position 20 or 24 is a Lys conjugated to a fatty diacid, or Tyr; X.sup.16 is aib, Ala, Ser, or Glu; X.sup.20 is Lys is conjugated to a fatty diacid, pAF conjugated to a fatty diacid, norleucine (Nle) conjugated to a fatty diacid, or Gln; X.sup.21 is Lys conjugated to a fatty diacid or pAF conjugated to a fatty diacid, Asp, alpha-methyl-L-phenylalanine (alpha-MF), or alpha-MD; X.sup.24 is Gln, Lys conjugated to a fatty diacid, or pAF conjugated to a fatty diacid; X.sup.25 is Trp or alpha-methyl-L-tryptophan (alpha-MW); X.sup.27 is L-Met sulphone or Leu; X.sup.28 is Glu, Asp, alpha-MD, Lys, aib, Ala, Lys conjugated to a fatty diacid or pAF conjugated to a fatty diacid; and X.sup.30 is absent or Lys linked at the C-terminus to gamma-Glu when X.sup.27 is Leu or L-Met-sulphone and X.sup.28 is Ala, aib, alpha-MD, or Lys conjugated to a fatty diacid; with the proviso that for each co-agonist peptide, only one of X.sup.10, X.sup.20, X.sup.21, X.sup.24, or X.sup.28 is conjugated to a fatty diacid.

20. A GCG/GLP-1 receptor co-agonist peptide wherein the GCG/GLP-1 receptor co-agonist peptide is selected from the group consisting of SEQ ID No. NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO: 18, SEQ ID NO: 19, SEQ ID NO: 20, SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO: 26, SEQ ID NO: 27, SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 31, SEQ ID NO: 32, SEQ ID NO: 33, SEQ ID NO: 34, SEQ ID NO: 35, SEQ ID NO: 36, SEQ ID NO: 37, SEQ ID NO: 38, SEQ ID NO: 39, SEQ ID NO: 40, SEQ ID NO: 41, SEQ ID NO: 42, SEQ ID NO: 43, SEQ ID NO: 44, SEQ ID NO: 45, SEQ ID NO: 47, SEQ ID NO: 48, SEQ ID NO: 49, SEQ ID NO: 50, SEQ ID NO: 51, SEQ ID NO: 52, SEQ ID NO: 54, SEQ ID NO: 55, SEQ ID NO: 56, SEQ ID NO: 57, SEQ ID NO: 58, SEQ ID NO: 64, SEQ ID NO: 65, SEQ ID NO: 66, SEQ ID NO: 67, SEQ ID NO: 68, SEQ ID NO: 69, SEQ ID NO: 70, SEQ ID NO: 71, SEQ ID NO: 72, SEQ ID NO: 73, SEQ ID NO: 74, SEQ ID NO: 75, SEQ ID NO: 76, SEQ ID NO: 77, SEQ ID NO: 78, SEQ ID NO: 79, SEQ ID NO: 80, SEQ ID NO: 81, SEQ ID NO: 82, SEQ ID NO: 83, SEQ ID NO: 84, SEQ ID NO: 85, SEQ ID NO: 86, SEQ ID NO: 87, SEQ ID NO: 88, SEQ ID NO: 89, SEQ ID NO: 90, SEQ ID NO: 91, SEQ ID NO: 92, SEQ ID NO: 93, SEQ ID NO: 94, SEQ ID NO: 95, SEQ ID NO: 96, SEQ ID NO: 97, SEQ ID NO: 98, SEQ ID NO: 99, SEQ ID NO: 100, SEQ ID NO: 101, SEQ ID NO: 102, SEQ ID NO: 103, and SEQ ID NO: 104, or a pharmaceutically acceptable salt or counterion thereof.

21. A GCG/GLP-1 receptor co-agonist peptide wherein the GCG/GLP-1 receptor co-agonist peptide is selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 10, SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO: 18, SEQ ID NO: 19, SEQ ID NO: 20, SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO: 26, SEQ ID NO: 27, SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 31, SEQ ID NO: 32, SEQ ID NO: 33, SEQ ID NO: 34, SEQ ID NO: 35, SEQ ID NO: 36, SEQ ID NO: 37, SEQ ID NO: 38, SEQ ID NO: 39, SEQ ID No. NO: SEQ ID NO: 41, SEQ ID NO: 42, SEQ ID NO: 43, SEQ ID NO: 44, SEQ ID NO: 45, SEQ ID NO: 47, SEQ ID NO: 48, SEQ ID NO: 49, SEQ ID NO: 50, SEQ ID NO: 51, SEQ ID NO: 52, SEQ ID NO: 53, SEQ ID NO: 54, SEQ ID NO: 55, SEQ ID NO: 56, SEQ ID NO: 57, SEQ ID NO: 58, SEQ ID NO: 64, SEQ ID NO: 65, SEQ ID NO: 66, SEQ ID NO: 67, SEQ ID NO: 68, SEQ ID NO: 69, SEQ ID NO: 70, SEQ ID NO: 71, SEQ ID NO: 72, SEQ ID NO: 73, SEQ ID NO: 74, SEQ ID NO: 75, SEQ ID NO: 76, SEQ ID NO: 77, SEQ NO: 78, SEQ ID NO: 79, SEQ ID NO: 80, SEQ ID NO: 81, SEQ ID NO: 82, SEQ ID NO: 83, SEQ ID NO: 84, SEQ ID NO: 85, SEQ ID NO: 86, SEQ ID NO: 87, SEQ ID NO: 88, SEQ ID NO: 89, SEQ ID NO: 90, SEQ ID NO: 91, SEQ ID NO: 92, SEQ ID NO: 93, SEQ ID NO: 94, SEQ ID NO: 95, SEQ ID NO: 96, SEQ ID NO: 97, SEQ ID NO: 98, SEQ ID NO: 99, SEQ ID NO: 100, SEQ ID NO: 101, SEQ ID NO: 102, SEQ ID NO: 103, and SEQ ID NO: 104, or a pharmaceutically acceptable salt or counterion thereof.

22. A GCG/GLP-1 receptor co-agonist peptide comprising the formula TABLE-US-00019 (SEQ ID NO: 106) HX.sup.2QGTFTSDX.sup.10SLYLDX.sup.16RAAX.sup.20X.sup.21FVX.sup.24X.sup.25LX.sup.27X.sup.28T-NH.sub.2 wherein X.sup.2 is alpha-aminoisobutyric acid (aib), D-Ser, or alpha-Methyl-L-Serine (alpha-MS); X.sup.9 is Asp or alpha-Methyl-L-Aspartic acid (alpha-MD); X.sup.10 is Lys or p-aminomethyl-L-phenylalanine (pAF) conjugated to a fatty acid provided that the amino acid at position 20 or 24 is a Lys conjugated to a fatty diacid, or Tyr; X.sup.16 is aib, Ala, Ser, or Glu; X.sup.20 is Lys is conjugated to a fatty diacid, pAF conjugated to a fatty diacid, norleucine (Nle) conjugated to a fatty diacid, or Gln; X.sup.21 is Lys conjugated to a fatty diacid, pAF conjugated to a fatty diacid, Asp, alpha-methyl-L-phenylalanine (alpha-MF), or alpha-MD; X.sup.24 is Gln, Lys conjugated to a fatty diacid, or pAF conjugated to a fatty diacid; X.sup.25 is Trp or alpha-methyl-L-tryptophan (alpha-MW); X.sup.27 is L-Met sulphone or Leu; and X.sup.28 is Glu, Asp, alpha-MD, Lys, aib, Ala, Lys conjugated to a fatty diacid or pAF conjugated to a fatty diacid; with the proviso that for each co-agonist peptide only one of X.sup.10, X.sup.20, X.sup.21, X.sup.24, or X.sup.28 is conjugated to a fatty diacid.

23. A GCG/GLP-1 receptor co-agonist peptide comprising the formula TABLE-US-00020 (SEQ ID NO: 107) HX.sup.2QGTFTSDX.sup.10SLYLDX.sup.16RAAX.sup.20X.sup.21FVX.sup.24X.sup.25LX.sup.27X.sup.28T-NH.sub.2 wherein X.sup.2 is alpha-aminoisobutyric acid (aib); X.sup.9 is Asp or alpha-Methyl-L-Aspartic acid (alpha-MD); X.sup.10 is Lys or p-aminomethyl-L-phenylalanine (pAF) conjugated to a fatty acid provided that the amino acid at position 20 or 24 is a Lys conjugated to a fatty diacid, or Tyr; X.sup.16 is aib, Ala, Ser, or Glu; X.sup.20 is Lys is conjugated to a fatty diacid, pAF conjugated to a fatty diacid, norleucine (Nle) conjugated to a fatty diacid, or Gln; X.sup.21 is Lys conjugated to a fatty diacid, pAF conjugated to a fatty diacid, Asp, alpha-methyl-L-phenylalanine (alpha-MF), or alpha-MD; X.sup.24 is Gln, Lys conjugated to a fatty diacid, or pAF conjugated to a fatty diacid; X.sup.25 is Trp or alpha-methyl-L-tryptophan (alpha-MW); X.sup.27 is L-Met sulphone or Leucine; X.sup.28 is Glu, Asp, alpha-MD, Lys, aib, Ala, Lys conjugated to a fatty diacid or pAF conjugated to a fatty diacid; and X.sup.30 is absent or Lys linked at the C-terminus to gamma-Glu when X.sup.27 is Leu or L-Met-sulphone, and X.sup.28 is Ala, aib, alpha-MD, or Lys conjugated to a fatty diacid; with the proviso that for each co-agonist peptide, only one of X.sup.10, X.sup.20, X.sup.21, X.sup.24, or X.sup.28 is conjugated to a fatty diacid and excludes peptides disclosed in Table 1 of WO2017074798.

24. A GCG/GLP-1 receptor co-agonist peptide wherein the GCG/GLP-1 receptor co-agonist peptide is selected from the group consisting of SEQ ID NO: 46, SEQ ID NO: 59, SEQ ID NO: 60, SEQ ID NO: 61, SEQ ID NO: 62, SEQ ID NO: 63, SEQ ID NO: 120, SEQ ID NO: 121, and SEQ ID NO: 122.

Description

DETAILED DESCRIPTION OF THE INVENTION

(1) The present invention provides glucagon analogs that display activity at the GLP-1 receptor (GLP-1) and the glucagon (GCG) receptor and that have a long-acting profile comprising a prolonged blood serum half-life. These GCG/GLP-1 receptor co-agonist peptides may have a blood serum half-life of at least one day, two days, three days, four days, five days, six days, or seven days.

(2) The GCG/GLP-1 receptor co-agonist peptides comprise (i) an amino acid substitution at position 2 of the peptide that confers resistance to dipeptidyl peptidase IV (DPPIV) degradation of the peptide, (ii) a lysine (Lys) at position 20, 21, 24, or 28 conjugated to a fatty diacid; or, p-aminomethyl-L-phenylalanine (pAF) at position 10, 20, 21, 24, or 28 conjugated to a fatty diacid; or, Norleucine (Nle) at position 20 conjugated to a fatty diacid, and (iii) one, two, three, four, five, or more amino acid substitutions and/or additions, which may control the relative activity of the GCG/GLP-1 receptor co-agonist peptide at the GLP-1 receptor verses the glucagon receptor or may confer enhanced biophysical stability and/or aqueous solubility to the GCG/GLP-1 co-agonist peptides.

(3) In particular aspects of the invention, (i) GCG/GLP-1 receptor co-agonist peptides are provided that have higher activity at the GCG receptor versus the GLP-1 receptor, (ii) GCG/GLP-1 receptor co-agonist peptides that have approximately equivalent activity at both receptors, and (iii) GCG/GLP-1 receptor co-agonist peptides that have higher activity at the GLP-1 receptor versus the glucagon receptor.

(4) The GCG/GLP-1 receptor co-agonist peptides disclosed herein are useful for the treatment of metabolic diseases or disorders. Such metabolic diseases or disorders, include but are not limited to, diabetes (e.g., Type 1 diabetes, Type 2 diabetes, or gestational diabetes), non-alcoholic fatty liver disease (NAFLD), non-alcoholic steatohepatitis (NASH), and obesity. In particular embodiments, the GCG/GLP-1 receptor co-agonist peptides may be useful for the simultaneous treatment of one or more of the aforementioned metabolic disorders.

(5) In one embodiment, the present invention provides a GCG/GLP-1 receptor co-agonist peptide comprising the formula

(6) TABLE-US-00008 (SEQ ID NO: 105) HX.sup.2QGTFTSDX.sup.10SLYLDX.sup.16RAAX.sup.20X.sup.21FVX.sup.24X.sup.25LX.sup.27X.sup.28TX.sup.30-NH.sub.2
wherein X.sup.2 is alpha-aminoisobutyric acid (aib), D-Ser, or alpha-Methyl-L-Serine (alpha-MS); X.sup.9 is Asp or alpha-Methyl-L-Aspartic acid (alpha-MD); X.sup.19 is Lys or p-aminomethyl-L-phenylalanine (pAF) conjugated to a fatty acid provided that the amino acid at position 20 or 24 is a Lys conjugated to a fatty diacid or Tyr; X.sup.16 is aib, Ala, Ser, or Glu; X.sup.20 is Lys is conjugated to a fatty diacid, pAF conjugated to a fatty diacid, norleucine (Nle) conjugated to a fatty diacid, or Gln; X.sup.21 is Lys conjugated to a fatty diacid or pAF conjugated to a fatty diacid, Asp, alpha-methyl-L-phenylalanine (alpha-MF), or alpha-MD; X.sup.24 is Gln, Lys conjugated to a fatty diacid, or pAF conjugated to a fatty diacid; X.sup.25 is Trp or alpha-methyl-L-tryptophan (alpha-MW); X.sup.27 is L-Met sulphone or Leucine; X.sup.28 is Glu, Asp or alpha-MD, Lys, aib, or Ala, Lys conjugated to a fatty diacid or pAF conjugated to a fatty diacid; and X.sup.39 is absent or Lys linked at the C-terminus to gamma-Glu when X.sup.27 is Leu or L-Met-sulphone and X.sup.28 is Ala, aib, alpha-MD, or Lys conjugated to a fatty diacid; with the proviso that for each co-agonist peptide, only one of X.sup.10, X.sup.20, X.sup.21, X.sup.24, or X.sup.28 is conjugated to a fatty diacid.

(7) In another embodiment, the present invention provides a GCG/GLP-1 receptor co-agonist peptide comprising the formula

(8) TABLE-US-00009 (SEQ ID NO: 123) HX.sup.2QGTFTSX.sup.9X.sup.10SLYLDX.sup.16RAAX.sup.20X.sup.21FVX.sup.24X.sup.25LX.sup.27X.sup.28TX.sup.30-NH.sub.2
or a pharmaceutically acceptable salt or counterion thereof, wherein
X.sup.2 is alpha-aminoisobutyric acid (aib), D-Ser, or alpha-Methyl-L-Serine (alpha-MS); X.sup.9 is Asp, or Glu; X.sup.10 is Lys or p-aminomethyl-L-phenylalanine (pAF) conjugated to a fatty acid provided that the amino acid at position 20 or 24 is a Lys conjugated to a fatty diacid or Tyr; X.sup.16 is aib, Ala, Ser, or Glu; X.sup.20 is Lys is conjugated to a fatty diacid, pAF conjugated to a fatty diacid, norleucine (Nle) conjugated to a fatty diacid, or Gln; X.sup.21 is Lys conjugated to a fatty diacid or pAF conjugated to a fatty diacid, Asp, alpha-methyl-L-phenylalanine (alpha-MF), or alpha-MD; X.sup.24 is Gln, Lys conjugated to a fatty diacid, or pAF conjugated to a fatty diacid; X.sup.25 is Trp or alpha-methyl-L-tryptophan (alpha-MW); X.sup.27 is L-Met sulphone or Leucine; X.sup.28 is Glu, Asp or alpha-MD, Lys, aib, or Ala, Lys conjugated to a fatty diacid or pAF conjugated to a fatty diacid; and X.sup.30 is absent or Lys linked at the C-terminus to gamma-Glu when X.sup.27 is Leu or L-Met-sulphone and X.sup.28 is Ala, aib, alpha-MD, or Lys conjugated to a fatty diacid; with the proviso that for each co-agonist peptide, only one of X.sup.10, X.sup.20, X.sup.21, X.sup.24, or X.sup.28 is conjugated to a fatty diacid. In a class of this embodiment, X.sup.9 is Asp, or Glu. In another class of this embodiment, X.sup.9 is Asp.

(9) In another embodiment, the present invention provides a GCG/GLP-1 receptor co-agonist peptide comprising the formula
HX.sup.2QGTFTSDX.sup.10SLYLDX.sup.16RAAX.sup.20X.sup.21FVX.sup.24X.sup.25.sub.LX.sup.27X.sup.28T-NH.sub.2  SEQ ID NO: 106
wherein X.sup.2 is alpha-aminoisobutyric acid (aib), D-Ser, or alpha-Methyl-L-Serine (alpha-MS); X.sup.9 is Asp or alpha-Methyl-L-Aspartic acid (alpha-MD); X.sup.10 is Lys or p-aminomethyl-L-phenylalanine (pAF) conjugated to a fatty acid provided that the amino acid at position 20 or 24 is a Lys conjugated to a fatty diacid or Tyr; X.sup.16 is aib, Ala, Ser, or Glu; X.sup.20 is Lys is conjugated to a fatty diacid, pAF conjugated to a fatty diacid, norleucine (Nle) conjugated to a fatty diacid, or Gln; X.sup.21 is Lys conjugated to a fatty diacid or pAF conjugated to a fatty diacid, Asp, alpha-methyl-L-phenylalanine (alpha-MF), or alpha-MD; X.sup.24 is Gln, Lys conjugated to a fatty diacid, or pAF conjugated to a fatty diacid; X.sup.25 is Trp or alpha-methyl-L-tryptophan (alpha-MW); X.sup.27 is L-Met sulphone or Leucine; X.sup.28 is Glu, Asp or alpha-MD, Lys, aib, or Ala, Lys conjugated to a fatty diacid or pAF conjugated to a fatty diacid; with the proviso that for each co-agonist peptide, only one of X.sup.10, X.sup.20, X.sup.21, X.sup.24, or X.sup.28 is conjugated to a fatty diacid.

(10) In another embodiment, the present invention provides a GCG/GLP-1 receptor co-agonist peptide comprising the formula

(11) TABLE-US-00010 (SEQ ID NO: 124) HX.sup.2QGTFTSX.sup.9X.sup.10SLYLDX.sup.16RAAX.sup.20X.sup.21FVX.sup.24X.sup.25LX.sup.27X.sup.28T-NH.sub.2
or a pharmaceutically acceptable salt or counterion thereof, wherein
X.sup.2 is alpha-aminoisobutyric acid (aib), D-Ser, or alpha-Methyl-L-Serine (alpha-MS); X.sup.9 is Asp, or Glu; X.sup.10 is Lys or p-aminomethyl-L-phenylalanine (pAF) conjugated to a fatty acid provided that the amino acid at position 20 or 24 is a Lys conjugated to a fatty diacid or Tyr; X.sup.16 is aib, Ala, Ser, or Glu; X.sup.20 is Lys is conjugated to a fatty diacid, pAF conjugated to a fatty diacid, norleucine (Nle) conjugated to a fatty diacid, or Gln; X.sup.21 is Lys conjugated to a fatty diacid or pAF conjugated to a fatty diacid, Asp, alpha-methyl-L-phenylalanine (alpha-MF), or alpha-MD; X.sup.24 is Gln, Lys conjugated to a fatty diacid, or pAF conjugated to a fatty diacid; X.sup.25 is Trp or alpha-methyl-L-tryptophan (alpha-MW); X.sup.27 is L-Met sulphone or Leucine; X.sup.28 is Glu, Asp or alpha-MD, Lys, aib, or Ala, Lys conjugated to a fatty diacid or pAF conjugated to a fatty diacid; with the proviso that for each co-agonist peptide, only one of X.sup.10, X.sup.20, X.sup.21, X.sup.24, or X.sup.28 is conjugated to a fatty diacid. In a class of this embodiment, X.sup.9 is Asp, or Glu. In another class of this embodiment, X.sup.9 is Asp.

(12) In another embodiment, the present invention provides a GCG/GLP-1 receptor co-agonist peptide comprising the formula

(13) TABLE-US-00011 (SEQ ID NO: 107) HX.sup.2QGTFTSDX.sup.10SLYLDX.sup.16RAAX.sup.20X.sup.21FVX.sup.24X.sup.25LX.sup.27X.sup.28T-NH.sub.2
wherein X.sup.2 is alpha-aminoisobutyric acid (aib); X.sup.9 is Asp or alpha-Methyl-L-Aspartic acid (alpha-MD); X.sup.10 is Lys or p-aminomethyl-L-phenylalanine (pAF) conjugated to a fatty acid provided that the amino acid at position 20 or 24 is a Lys conjugated to a fatty diacid or Tyr; X.sup.16 is aib, Ala, Ser, or Glu; X.sup.20 is Lys is conjugated to a fatty diacid, pAF conjugated to a fatty diacid, norleucine (Nle) conjugated to a fatty diacid, or Gln; X.sup.21 is Lys conjugated to a fatty diacid or pAF conjugated to a fatty diacid, Asp, alpha-methyl-L-phenylalanine (alpha-MF), or alpha-MD; X.sup.24 is Gln, Lys conjugated to a fatty diacid, or pAF conjugated to a fatty diacid; X.sup.25 is Trp or alpha-methyl-L-tryptophan (alpha-MW); X.sup.27 is L-Met sulphone or Leucine; X.sup.28 is Glu, Asp or alpha-MD, Lys, aib, or Ala, Lys conjugated to a fatty diacid or pAF conjugated to a fatty diacid; and X.sup.30 is absent or Lys linked at the C-terminus to gamma-Glu when X.sup.27 is Leu or L-Met-sulphone and X.sup.28 is Ala, aib, alpha-MD, or Lys conjugated to a fatty diacid; with the proviso that for each co-agonist peptide, only one of X.sup.10, X.sup.20, X.sup.21, X.sup.24, or X.sup.28 is conjugated to a fatty diacid.

(14) In another embodiment, the present invention provides a GCG/GLP-1 receptor co-agonist peptide comprising the formula

(15) TABLE-US-00012 (SEQ ID NO: 125) HX.sup.2QGTFTSX.sup.9X.sup.10SLYLDX.sup.16RAAX.sup.20X.sup.21FVX.sup.24X.sup.25LX.sup.27X.sup.28T-NH.sub.2
or a pharmaceutically acceptable salt or counterion thereof, wherein
X.sup.2 is alpha-aminoisobutyric acid (aib); X.sup.9 is Asp, or Glu; X.sup.10 is Lys or p-aminomethyl-L-phenylalanine (pAF) conjugated to a fatty acid provided that the amino acid at position 20 or 24 is a Lys conjugated to a fatty diacid or Tyr; X.sup.16 is aib, Ala, Ser, or Glu; X.sup.20 is Lys is conjugated to a fatty diacid, pAF conjugated to a fatty diacid, norleucine (Nle) conjugated to a fatty diacid, or Gln; X.sup.21 is Lys conjugated to a fatty diacid or pAF conjugated to a fatty diacid, Asp, alpha-methyl-L-phenylalanine (alpha-MF), or alpha-MD; X.sup.24 is Gln, Lys conjugated to a fatty diacid, or pAF conjugated to a fatty diacid; X.sup.25 is Trp or alpha-methyl-L-tryptophan (alpha-MW); X.sup.27 is L-Met sulphone or Leucine; X.sup.28 is Glu, Asp or alpha-MD, Lys, aib, or Ala, Lys conjugated to a fatty diacid or pAF conjugated to a fatty diacid; and X.sup.30 is absent or Lys linked at the C-terminus to gamma-Glu when X.sup.27 is Leu or L-Met-sulphone and X.sup.28 is Ala, aib, alpha-MD, or Lys conjugated to a fatty diacid; with the proviso that for each co-agonist peptide, only one of X.sup.10, X.sup.20, X.sup.21, X.sup.24, or X.sup.28 is conjugated to a fatty diacid. In a class of this embodiment, X.sup.9 is Asp, or Glu. In another class of this embodiment, X.sup.9 is Asp.

(16) In another embodiment, the GCG/GLP-1 receptor co-agonist peptide selected from the group consisting of TP565, TP579, TP583, TP584, TP578, TP580, TP581, TP582, TP588, TP589, TP590, TP592, TP594, TP576, TP577, TP586, TP587, TP591, TP593, TP595, TP596, TP599, TP600, TP601, TP602, TP603, TP605, TP606, TP607, TP610, TP611, TP612, TP613, TP614, TP615, TP616, TP617, TP618, TP619, TP620, TP621, TP622, TP623, TP624, TP625, TP626, TP627, TP629, TP631, TP632, TP633, TP634, TP635, TP636, TP637, TP638, TP639, TP657, TP658, TP659, TP660, TP661, TP662, TP663, TP664, TP665, TP666, TP667, TP672, TP673, TP674, TP675, TP676, TP677, TP678, TP679, TP680, TP681, TP682, TP683, TP685, TP693, TP699, TP700, TP701, TP702, TP703, TP704, TP705, TP712, TP713, TP735, TP736, TP737, TP811, TP812, TP813, TP814, TP815, TP825, TP826, TP827, and TP828, or a pharmaceutically acceptable salt or counterion thereof.

(17) In another embodiment, the GCG/GLP-1 receptor co-agonist peptide selected from the group consisting of TP565, TP579, TP583, TP584, TP578, TP580, TP581, TP582, TP588, TP589, TP590, TP592, TP594, TP576, TP577, TP586, TP587, TP591, TP593, TP595, TP596, TP599, TP600, TP601, TP602, TP603, TP605, TP606, TP607, TP610, TP611, TP612, TP613, TP614, TP615, TP616, TP617, TP618, TP619, TP620, TP621, TP622, TP623, TP624, TP625, TP626, TP627, TP629, TP631, TP632, TP633, TP634, TP635, TP636, TP637, TP638, TP639, TP657, TP658, TP659, TP660, TP661, TP662, TP663, TP664, TP665, TP666, TP667, TP672, TP673, TP674, TP675, TP676, TP677, TP678, TP679, TP680, TP681, TP682, TP683, TP685, TP693, TP700, TP701, TP702, TP704, TP705, TP712, TP713, TP735, TP736, TP737, TP811, TP812, TP813, TP814, TP815, TP825, TP826, TP827, and TP828, or a pharmaceutically acceptable salt or counterion thereof.

(18) In another embodiment, the GCG/GLP-1 receptor co-agonist peptide selected from the group consisting of TP699, and TP703, or a pharmaceutically acceptable salt or counterion thereof.

(19) In another embodiment, the present invention provides a GCG/GLP-1 receptor co-agonist peptide comprising the formula

(20) TABLE-US-00013 (SEQ ID NO: 108) HX.sup.2QGTFX.sup.7SX.sup.9X.sup.10SX.sup.12X.sup.13X.sup.14X.sup.15X.sup.16X.sup.17AX.sup.19X.sup.20X.sup.21X.sup.22X.sup.23X.sup.24X.sup.25LX.sup.27X.sup.28 X.sup.29
or a pharmaceutically acceptable salt or counterion thereof, wherein X.sup.2 is alpha-aminoisobutyric acid (aib), D-Ser, or alpha-Methyl-L-Serine (αMS); X.sup.7 is Thr, Phe or Leu; X.sup.9 is Asp or Glu; X.sup.10 is Tyr, norleucine (Nle) conjugated to a fatty acid, p-aminomethyl-L-phenylalanine (pAF) conjugated to a fatty diacid, Lys conjugated to a fatty acid provided that the amino acid at position 20 or 24 is a Lys conjugated to a fatty diacid, or p-aminomethyl-L-phenylalanine (pAF) conjugated to a fatty acid provided that the amino acid at position 24 is a Lys conjugated to a fatty diacid; X.sup.12 is Lys, or Glu; X.sup.13 is Tyr, Leu, or Lys; X.sup.14 is Leu, or Asp; X.sup.15 is Asp, Glu, alpha-Methyl-L-Aspartic acid (αMD), or alpha-aminoisobutyric acid (aib); X.sup.16 is alpha-aminoisobutyric acid (aib), Ala, Glu, Ser, Arg, or Lys; X.sup.17 is Arg, Lys, Leu, or Ala; X.sup.19 is Ala, or Gln; X.sup.20 is Gln, Lys, Lys conjugated to a fatty diacid, p-aminomethyl-L-phenylalanine (pAF) conjugated to a fatty diacid, or norleucine (Nle) conjugated to a fatty diacid; X.sup.21 is Asp, Phe, Glu, alpha-Methyl-L-Aspartic acid (αMD), Lys conjugated to a fatty diacid, or pAF conjugated to a fatty diacid; X.sup.22 is Phe, Val, or alpha-methyl-L-phenylalanine (αMF); X.sup.23 is Val, or Gln; X.sup.24 is Gln, Glu, Lys conjugated to a fatty diacid, or pAF conjugated to a fatty diacid; X.sup.25 is Trp, or alpha-methyl-L-tryptophan (αMW); X.sup.27 is L-methionine sulphone (2), or Leu; X.sup.28 is Asp, alpha-Methyl-L-Aspartic acid (αMD), alpha-aminoisobutyric acid (aib), Ala, Lys, Gln, Glu, γ-glutamic acid (γE), Lys conjugated to a fatty diacid, or pAF conjugated to a fatty diacid; and X.sup.29 is Thr-OH, Thr-NH.sub.2, or Thr(Lys-γ-glutamic acid)NH.sub.2; with the proviso that for each co-agonist peptide, only one or two of X.sup.10, X.sup.20, X.sup.21, X.sup.24, or X.sup.28 are conjugated to a fatty diacid.

(21) In one embodiment, with the proviso that for each co-agonist peptide, one of X.sup.10, X.sup.20, X.sup.21, X.sup.24, or X.sup.28 is conjugated to a fatty diacid. In another embodiment, with the proviso that for each co-agonist peptide, two of X.sup.10, X.sup.20, X.sup.21, X.sup.24, or X.sup.28 are conjugated to a fatty diacid.

(22) In another embodiment, X.sup.2 is alpha-aminoisobutyric acid (aib), or alpha-Methyl-L-Serine (αMS). In another embodiment, X.sup.2 is alpha-aminoisobutyric acid (aib). In another embodiment, X.sup.2 is alpha-Methyl-L-Serine (αMS). In another embodiment, X.sup.2 is D-Ser.

(23) In another embodiment, X.sup.7 is Thr. In another embodiment, X.sup.7 is Phe. In another embodiment, X.sup.7 is Leu.

(24) In another embodiment, X.sup.9 is Asp. In another embodiment, X.sup.9 is Glu.

(25) In another embodiment, X.sup.10 is norleucine (Nle) conjugated to a fatty acid, p-aminomethyl-L-phenylalanine (pAF) conjugated to a fatty diacid, Lys conjugated to a fatty acid provided that the amino acid at position 20 or 24 is a Lys conjugated to a fatty diacid, or p-aminomethyl-L-phenylalanine (pAF) conjugated to a fatty acid provided that the amino acid at position 24 is a Lys conjugated to a fatty diacid. In another embodiment, X.sup.10 is norleucine (Nle) conjugated to a fatty acid, p-aminomethyl-L-phenylalanine (pAF) conjugated to a fatty diacid, Lys conjugated to a fatty acid provided that the amino acid at position 20 or 24 is a Lys conjugated to a fatty diacid, or p-aminomethyl-L-phenylalanine (pAF) conjugated to a fatty acid provided that the amino acid at position 24 is a Lys conjugated to a fatty diacid. In another embodiment, X.sup.10 is p-aminomethyl-L-phenylalanine (pAF) conjugated to a fatty diacid, Lys conjugated to a fatty acid provided that the amino acid at position 20 or 24 is a Lys conjugated to a fatty diacid, or p-aminomethyl-L-phenylalanine (pAF) conjugated to a fatty acid provided that the amino acid at position 24 is a Lys conjugated to a fatty diacid. In another embodiment, X.sup.10 is p-aminomethyl-L-phenylalanine (pAF) conjugated to a fatty diacid. In another embodiment, X.sup.10 is p-aminomethyl-L-phenylalanine (pAF) conjugated to a fatty acid provided that the amino acid at position 24 is a Lys conjugated to a fatty diacid. In another embodiment, X.sup.10 is Lys conjugated to a fatty acid provided that the amino acid at position 20 or 24 is a Lys conjugated to a fatty diacid, or p-aminomethyl-L-phenylalanine (pAF) conjugated to a fatty acid provided that the amino acid at position 24 is a Lys conjugated to a fatty diacid. In another embodiment, X.sup.10 is Tyr.

(26) In another embodiment, X.sup.12 is Lys. In another embodiment, X.sup.12 is Glu.

(27) In another embodiment, X.sup.13 is Tyr, or Leu. In another embodiment, X.sup.13 is Tyr. In another embodiment, X.sup.13 is Leu. In another embodiment, X.sup.13 is Lys.

(28) In another embodiment, X.sup.14 is Leu. In another embodiment, X.sup.14 is Asp.

(29) In another embodiment, X.sup.15 is Asp. In another embodiment, X.sup.15 is Glu. In another embodiment, X.sup.15 is □MD. In another embodiment, X.sup.15 is alpha-aminoisobutyric acid (aib).

(30) In another embodiment, X.sup.16 is alpha-aminoisobutyric acid (aib), or Ala. In another embodiment, X.sup.16 is alpha-aminoisobutyric acid (aib). In another embodiment, X.sup.16 is Ala. In another embodiment, X.sup.16 is Glu. In another embodiment, X.sup.16 is Ser. In another embodiment, X.sup.16 is Arg. In another embodiment, X.sup.16 is Lys.

(31) In another embodiment, X.sup.17 is Arg. In another embodiment, X.sup.17 is Lys. In another embodiment, X.sup.17 is Leu. In another embodiment, X.sup.17 is Ala.

(32) In another embodiment, X.sup.19 is Ala. In another embodiment, X.sup.19 is Gln.

(33) In another embodiment, X.sup.20 is Gln, Lys conjugated to a fatty diacid, or norleucine (Nle) conjugated to a fatty diacid. In another embodiment, X.sup.20 is Gln. In another embodiment, X.sup.20 is Lys. In another embodiment, X.sup.20 is pAF conjugated to a fatty diacid. In another embodiment, X.sup.20 is Lys conjugated to a fatty diacid. In another embodiment, X.sup.20 is norleucine (Nle) conjugated to a fatty diacid.

(34) In another embodiment, X.sup.21 is Asp, Glu, Lys conjugated to a fatty diacid, or pAF conjugated to a fatty diacid. In another embodiment, X.sup.21 is Asp. In another embodiment, X.sup.21 is Glu. In another embodiment, X.sup.21 is Phe. In another embodiment, X.sup.21 is □MD. In another embodiment, X.sup.21 is Lys conjugated to a fatty diacid. In another embodiment, X.sup.21 is pAF conjugated to a fatty diacid.

(35) In another embodiment, X.sup.22 is Phe. In another embodiment, X.sup.22 is □MF. In another embodiment, X.sup.22 is Val.

(36) In another embodiment, X.sup.23 is Val. In another embodiment, X.sup.23 is Gln.

(37) In another embodiment, X.sup.24 is Gln. In another embodiment, X.sup.24 is Lys conjugated to a fatty diacid. In another embodiment, X.sup.24 is K(PEG.sub.2PEG.sub.2γEC.sub.18—OH). In another embodiment, X.sup.24 is pAF conjugated to a fatty diacid. In another embodiment, X.sup.24 is Glu.

(38) In another embodiment, X.sup.25 is Trp. In another embodiment, X.sup.25 is alpha-methyl-L-tryptophan (αMW).

(39) In another embodiment, X.sup.27 is L-methionine sulphone (2). In another embodiment, X.sup.27 is Leu.

(40) In another embodiment, X.sup.28 is Asp, alpha-Methyl-L-Aspartic acid (αMD), Glu, or pAF conjugated to a fatty diacid. In another embodiment, X.sup.28 is Asp. In another embodiment, X.sup.28 is alpha-Methyl-L-Aspartic acid (αMD). In another embodiment, X.sup.28 is Glu. In another embodiment, X.sup.28 is pAF conjugated to a fatty diacid.

(41) In another embodiment, X.sup.28 is alpha-aminoisobutyric acid (aib), Ala, Lys, □-Glu, or Lys conjugated to a fatty diacid. In another embodiment, X.sup.28 is alpha-aminoisobutyric acid (aib). In another embodiment, X.sup.28 is Ala. In another embodiment, X.sup.28 is Lys. In another embodiment, X.sup.28 is □-Glu. In another embodiment, X.sup.28 is Lys conjugated to a fatty diacid.

(42) In another embodiment, X.sup.29 is Thr-OH. In another embodiment, X.sup.29 is Thr-NH.sub.2. In another embodiment, X.sup.29 is Thr(Lys-γ-glutamic acid)NH.sub.2. In another embodiment, X.sup.29 is Thr-OH, or Thr-NH.sub.2. In another embodiment, X.sup.29 is Thr-OH, or Thr(Lys-γ-glutamic acid)NH.sub.2.

(43) In another embodiment, X.sup.29 is Thr-NH.sub.2, or Thr(Lys-γ-glutamic acid)NH.sub.2.

(44) In another embodiment, the fatty diacid comprises a C.sub.16, C.sub.18, or C.sub.20 fatty diacid.

(45) In another embodiment, the fatty diacid conjugated to the Lys or pAF via a gamma-Glu linker.

(46) In another embodiment, the fatty diacid conjugated to the Lys or pAF via a gamma-Glu linker, a PEG.sub.2 gamma-Glu linker, a PEG.sub.5 gamma-Glu linker, a PEG.sub.2PEG.sub.2-gamma-Glu linker, a PEG.sub.2 PEG.sub.2PEG.sub.2 gamma-Glu linker, wherein PEG.sub.2 is 8-amino-3,6-dioxaoctanoic acid, and PEG.sub.5 is 1-hydroxy-3,6,9,12,15-pentaoxaoctadecan-18-oic acid.

(47) In another embodiment, the fatty diacid conjugated to the Lys via a PEG.sub.2 gamma-Glu linker, a PEG.sub.5 gamma-Glu linker, a PEG.sub.2PEG.sub.2-gamma-Glu linker, a PEG.sub.2 PEG.sub.2PEG.sub.2 gamma-Glu linker, wherein PEG.sub.2 is 8-amino-3,6-dioxaoctanoic acid, and PEG.sub.5 is 1-hydroxy-3,6,9,12,15-pentaoxaoctadecan-18-oic acid.

(48) In another embodiment, the fatty diacid conjugated to the pAF via a gamma-Glu linker, a PEG.sub.2 gamma-Glu linker, a PEG.sub.2PEG.sub.2-gamma-Glu linker, a PEG.sub.2 PEG.sub.2PEG.sub.2 gamma-Glu linker, wherein PEG.sub.2 is 8-amino-3,6-dioxaoctanoic acid, and PEG.sub.5 is 1-hydroxy-3,6,9,12,15-pentaoxaoctadecan-18-oic acid.

(49) In another embodiment, the fatty diacid conjugated to the Lys or pAF via a PEG.sub.5 gamma-Glu linker, a PEG.sub.2PEG.sub.2-gamma-Glu linker, or a PEG.sub.2 PEG.sub.2PEG.sub.2 gamma-Glu linker, wherein PEG.sub.2 is 8-amino-3,6-dioxaoctanoic acid, and PEG.sub.5 is 1-hydroxy-3,6,9,12,15-pentaoxaoctadecan-18-oic acid.

(50) In another embodiment, the fatty diacid conjugated to the Lys via a PEG.sub.5 gamma-Glu linker, a PEG.sub.2PEG.sub.2-gamma-Glu linker, or a PEG.sub.2 PEG.sub.2PEG.sub.2 gamma-Glu linker, wherein PEG.sub.2 is 8-amino-3,6-dioxaoctanoic acid, and PEG.sub.5 is 1-hydroxy-3,6,9,12,15-pentaoxaoctadecan-18-oic acid.

(51) In another embodiment, the fatty diacid conjugated to the pAF via a PEG.sub.2PEG.sub.2-gamma-Glu linker, wherein PEG.sub.2 is 8-amino-3,6-dioxaoctanoic

(52) In another embodiment, the fatty acid comprises a C.sub.16 fatty acid.

(53) In another embodiment, the fatty acid conjugated to the Lys or pAF via a gamma-Glu-gamma-Glu linker.

(54) In another embodiment, the fatty acid conjugated to the Lys or pAF via a gamma-Glu linker.

(55) In another embodiment, the fatty acid comprises a C.sub.18 fatty diacid.

(56) In another embodiment, the fatty diacid conjugated to the norleucine (Nle) via a triazole Peg.sub.2Peg.sub.2-gamma-Glu linker.

(57) In another embodiment, the fatty acid comprises a C.sub.15, or C.sub.16 fatty acid.

(58) In another embodiment, the fatty acid conjugated to the norleucine (Nle) via a triazole linker, via a triazole gamma-Glu linker, or via a triazole-C.sub.4alkyl-Lys linker.

(59) In another embodiment, with the proviso that for each co-agonist peptide, one or two of X.sup.10, X.sup.20, X.sup.21, X.sup.24, or X.sup.28 is conjugated to a fatty diacid and excludes peptides disclosed in Table 1 of WO2017074798.

(60) In another embodiment, with the proviso that for each co-agonist peptide, two of X.sup.10, X.sup.20, X.sup.21, X.sup.24, or X.sup.28 is conjugated to a fatty diacid and excludes peptides disclosed in Table 1 of WO2017074798.

(61) In another embodiment, with the proviso that for each co-agonist peptide, only one of X.sup.10, X.sup.20, X.sup.21, X.sup.24, or X.sup.28 is conjugated to a fatty diacid and excludes peptides disclosed in Table 1 of WO2017074798.

(62) In another embodiment, the GCG/GLP-1 receptor co-agonist peptide of claim 1, wherein the GCG/GLP-1 receptor co-agonist peptide is selected from the group consisting of TP564, TP565, TP579, TP583, TP584, TP575, TP578, TP580, TP581, TP582, TP585, TP588, TP589, TP590, TP592, TP594, TP576, TP577, TP586, TP587, TP591, TP593, TP595, TP596, TP597, TP598, TP599, TP600, TP601, TP602, TP603, TP604, TP605, TP443, TP606, TP607, TP608, TP609, TP610, TP611, TP612, TP613, TP614, TP615, TP616, TP617, TP618, TP619, TP620, TP621, TP622, TP623, TP624, TP625, TP626, TP627, TP628, TP629, TP630, TP631, TP632, TP633, TP635, TP636, TP637, TP638, TP639, TP640, TP657, TP658, TP659, TP660, TP661, TP662, TP663, TP664, TP665, TP666, TP667, TP672, TP673, TP674, TP675, TP676, TP677, TP678, TP679, TP680, TP681, TP682, TP683, TP685, TP693, TP699, TP700, TP701, TP702, TP703, TP704, TP705, TP712, TP713, TP735, TP736, TP737, TP811, TP812, TP813, TP814, TP815, TP825, TP826, TP827, TP828, TP829, TP830, and TP831, or a pharmaceutically acceptable salt or counterion thereof.

(63) In another embodiment, the GCG/GLP-1 receptor co-agonist peptide selected from the group consisting of TP699, and TP703, or a pharmaceutically acceptable salt or counterion thereof.

(64) In another embodiment, the GCG/GLP-1 receptor co-agonist peptide of claim 1, wherein the GCG/GLP-1 receptor co-agonist peptide is selected from the group consisting of TP575, TP597, TP604, TP608, TP609, TP615, TP617, TP630, TP640, TP672, TP676, TP680, TP704, TP712, and TP813, or a pharmaceutically acceptable salt or counterion thereof.

(65) In another embodiment, the GCG/GLP-1 receptor co-agonist peptide of claim 1, wherein the GCG/GLP-1 receptor co-agonist peptide is TP575, or a pharmaceutically acceptable salt or counterion thereof. In another embodiment, the GCG/GLP-1 receptor co-agonist peptide of claim 1, wherein the GCG/GLP-1 receptor co-agonist peptide is TP597, or a pharmaceutically acceptable salt or counterion thereof. In another embodiment, the GCG/GLP-1 receptor co-agonist peptide of claim 1, wherein the GCG/GLP-1 receptor co-agonist peptide is TP604, or a pharmaceutically acceptable salt or counterion thereof. In another embodiment, the GCG/GLP-1 receptor co-agonist peptide of claim 1, wherein the GCG/GLP-1 receptor co-agonist peptide is TP608, or a pharmaceutically acceptable salt or counterion thereof. In another embodiment, the GCG/GLP-1 receptor co-agonist peptide of claim 1, wherein the GCG/GLP-1 receptor co-agonist peptide is TP609, or a pharmaceutically acceptable salt or counterion thereof. In another embodiment, the GCG/GLP-1 receptor co-agonist peptide of claim 1, wherein the GCG/GLP-1 receptor co-agonist peptide is TP615, or a pharmaceutically acceptable salt or counterion thereof. In another embodiment, the GCG/GLP-1 receptor co-agonist peptide of claim 1, wherein the GCG/GLP-1 receptor co-agonist peptide is TP617, or a pharmaceutically acceptable salt or counterion thereof. In another embodiment, the GCG/GLP-1 receptor co-agonist peptide of claim 1, wherein the GCG/GLP-1 receptor co-agonist peptide is TP630, or a pharmaceutically acceptable salt or counterion thereof. In another embodiment, the GCG/GLP-1 receptor co-agonist peptide of claim 1, wherein the GCG/GLP-1 receptor co-agonist peptide is TP640, or a pharmaceutically acceptable salt or counterion thereof. In another embodiment, the GCG/GLP-1 receptor co-agonist peptide of claim 1, wherein the GCG/GLP-1 receptor co-agonist peptide is TP672, or a pharmaceutically acceptable salt or counterion thereof. In another embodiment, the GCG/GLP-1 receptor co-agonist peptide of claim 1, wherein the GCG/GLP-1 receptor co-agonist peptide is TP676, or a pharmaceutically acceptable salt or counterion thereof. In another embodiment, the GCG/GLP-1 receptor co-agonist peptide of claim 1, wherein the GCG/GLP-1 receptor co-agonist peptide is TP680, or a pharmaceutically acceptable salt or counterion thereof. In another embodiment, the GCG/GLP-1 receptor co-agonist peptide of claim 1, wherein the GCG/GLP-1 receptor co-agonist peptide is TP704, or a pharmaceutically acceptable salt or counterion thereof. In another embodiment, the GCG/GLP-1 receptor co-agonist peptide of claim 1, wherein the GCG/GLP-1 receptor co-agonist peptide is TP712, or a pharmaceutically acceptable salt or counterion thereof. In another embodiment, the GCG/GLP-1 receptor co-agonist peptide of claim 1, wherein the GCG/GLP-1 receptor co-agonist peptide is TP813, or a pharmaceutically acceptable salt or counterion thereof.

(66) The aforementioned GCG/GLP-1 receptor co-agonist peptide represented by SEQ ID NO: 105, 106, 107, 108, 123, 124 and 125 excludes peptides disclosed in Table 1 of WO2017074798.

(67) In particular aspects, the fatty diacid comprises a C14, C15, C16, C17, C18, C19, or C20 fatty diacid. In a further aspect, the fatty diacid comprises a C16 or C18 fatty diacid.

(68) In particular aspects, the GCG/GLP-1 receptor co-agonist peptide comprises the fatty diacid conjugated to the Lys or pAF via a gamma-Glu, gamma-Glu linker.

(69) In particular aspects, the GCG/GLP-1 receptor co-agonist peptide comprises the fatty diacid conjugated to the Lys or pAF via a PEG.sub.2PEG.sub.2-gamma-Glu linker wherein PEG.sub.2 is 8-amino-3,6-dioxaoctanoic acid.

(70) In particular aspects, the GCG/GLP-1 receptor co-agonist peptide comprises at X.sup.10 a pAF conjugated to a fatty diacid.

(71) In particular aspects, the GCG/GLP-1 receptor co-agonist peptide comprises at X.sup.10 a Lys conjugated to a C16 fatty acid and a Lys at position 20 or 24 conjugated to a fatty diacid.

(72) In particular aspects, the GCG/GLP-1 receptor co-agonist peptide comprises at X.sup.20 a pAF conjugated to a fatty diacid or a Lys conjugated to a fatty diacid.

(73) In particular aspects, the GCG/GLP-1 receptor co-agonist peptide comprises at X.sup.21 a pAF conjugated to a fatty diacid or a Lys conjugated to a fatty diacid.

(74) In particular aspects, the GCG/GLP-1 receptor co-agonist peptide comprises at X.sup.24 a pAF conjugated to a fatty diacid or a Lys conjugated to a fatty diacid.

(75) In particular aspects, the GCG/GLP-1 receptor co-agonist peptide comprises at X.sup.28 a pAF conjugated to a fatty diacid or a Lys conjugated to a fatty diacid.

(76) Exemplary GCG/GLP-1 receptor co-agonist peptides within the scope of the invention are disclosed in Table 1.

(77) TABLE-US-00014 TABLE 1 SEQ ID NO: ID SEQUENCE   1 TP565 HUQGTFTSDYSKYLDARAAK(PEG.sub.2PEG.sub.2γEC.sub.20-OH)DFVQWL.sub.2αMDT-NH.sub.2   2 TP579 HsQGTFTSDYSKYLEURAAK(PEG.sub.2PEG.sub.2γEC.sub.20-OH)DFVQWLLUTKγE-NH.sub.2   3 TP583 HUQGTFTSDYSKYLDARAAQDFVpAF(PEG.sub.2PEG.sub.2γEC.sub.18-OH)WLLDT-NH.sub.2   4 TP584 HUQGTFTSDYSKYLDARAAQDFVpAF(PEG.sub.2PEG.sub.2γEC.sub.20-OH)WLLDT-NH.sub.2   5 TP578 HsQGTFTSDYSKYLDURAAK(PEG.sub.2PEG.sub.2γEC.sub.20-OH)DFVQWL2DT-NH.sub.2   6 TP580 HUQGTFTSDpAF(PEG.sub.2PEG.sub.2γEC.sub.20-OH)SKYLDARAAQDFVQWL2DT-NH.sub.2   7 TP581 HUQGTFTSDpAF(PEG.sub.2PEG.sub.2γEC.sub.20-OH)SKYLDURAAQDFVQWLLATKγE-NH.sub.2   8 TP582 HUQGTFTSDpAF(PEG.sub.2PEG.sub.2γEC.sub.20-OH)SKYLDURAAQDFVQWLLATKγE-NH.sub.2   9 TP585 HUQGTFTSDYSKYLDARAAQpAF(PEG.sub.2PEG.sub.2γEC.sub.20-OH)FVQWLLDT-NH.sub.2  10 TP588 HUQGTFTSDYSKYLDURAAK(PEG.sub.2PEG.sub.2γEC.sub.20-OH)DFVQWL2αMDT-NH.sub.2  11 TP589 HUQGTFTSDYSKYLDERAAK(PEG.sub.2PEG.sub.2γEC.sub.20-OH)DFVQWL2αMDT-NH.sub.2  12 TP590 HUQGTFTSDYSKYLDARAAK(PEG.sub.2PEG.sub.2γEC.sub.20-OH)DαMFVQWL2DT-NH.sub.2  13 TP592 HαMSQGTFTSDYSKYLDURAAK(PEG.sub.2PEG.sub.2γEC.sub.20-OH)DFVQWL2DT-NH.sub.2  14 TP594 HUQGTFTSDYSKYLDARAAK(PEG.sub.2PEG.sub.2γEC.sub.20-OH)αMDFVQWL2DT-NH.sub.2  15 TP576 HUQGTFTSDYSKYLDURAAQDFVK(PEG.sub.2PEG.sub.2γEC.sub.20-OH)WL2DT-NH.sub.2  16 TP577 HUQGTFTSDYSKYLDSRAAK(PEG.sub.2PEG.sub.2γEC.sub.20-OH)DFVQWL2DT-NH.sub.2  17 TP586 HUQGTFTSDYSKYLDERAApAF(PEG.sub.2PEG.sub.2γEC.sub.20-OH)DFVQWLLKT-NH.sub.2  18 TP587 HUQGTFTSDYSKYLDERAApAF(PEG.sub.2PEG.sub.2γEC.sub.18-OH)DFVQWLLKT-NH.sub.2  19 TP591 HUQGTFTSDYSKYLDARAAK(PEG.sub.2PEG.sub.2γEC.sub.18-OH)DαMFVQWL2DT-NH.sub.2  20 TP593 HαMSQGTFTSDYSKYLDURAAK(PEG.sub.2PEG.sub.2γEC.sub.20-OH)DFVQWL2DT-NH.sub.2  21 TP595 HUQGTFFSDYSKYLDARAAK(PEG.sub.2PEG.sub.2γEC.sub.18-OH)DFVQWL2DT-NH.sub.2  22 TP596 HUQGTFLSDYSKYLDARAAK(PEG.sub.2PEG.sub.2γEC.sub.18-OH)DFVQWL2DT-NH.sub.2  23 TP599 HUQGTFTSDYSKYLDALAAK(PEG.sub.2PEG.sub.2γEC.sub.18-OH)DFVQWL2DT-NH.sub.2  24 TP600 HUQGTFTSDYSKYLDARAAK(PEG.sub.2PEG.sub.2γEC.sub.18-OH)DFVQWL2QT-NH.sub.2  25 TP601 HUQGTFTSDK(γEγEC16)SKYLDARAAK(PEG.sub.2PEG.sub.2γEC.sub.18-OH)DFVQWL2DT-NH.sub.2  26 TP602 HUQGTFTSDYSKYLDARAAQDFVQWL2K(PEG.sub.2PEG.sub.2γEC.sub.18-OH)T-NH.sub.2  27 TP603 HUQGTFTSDYSKYLDARAAQDFVQWL2K(PEG.sub.2PEG.sub.2γEC.sub.20-OH)T-NH.sub.2  28 TP605 HUQGTFTSDYSKYLDARAAQDFVQWL2pAF(PEG.sub.2PEG.sub.2γEC.sub.20-OH)T-NH.sub.2  29 TP606 HUQGTFTSDK(γEγEC16)SKYLDARAAK(PEG.sub.2PEG.sub.2γEC.sub.16-OH)DFVQWL2DT-NH.sub.2  30 TP607 HUQGTFTSDK(γEγEC16)SKYLDARAAQDFVK(PEG.sub.2PEG.sub.2γEC.sub.16-OH)WL2αMDT-NH.sub.2  31 TP610 HUQGTFTSDYSKYLαMDARAAK(PEG.sub.2PEG.sub.2γEC.sub.18-OH)DFVQWL2αMDT-NH.sub.2  32 TP611 HUQGTFTSDYSKYLDARAAK(PEG.sub.2PEG.sub.2γEC.sub.18-OH)αMDFVQWL2αMDT-NH.sub.2  33 TP612 HUQGTFTSDYSKYLαMDARAAK(PEG.sub.2PEG.sub.2γEC.sub.18-OH)αMDFVQWL2αMDT-NH.sub.2  34 TP613 HUQGTFTSDYSKYLDERAAK(PEG.sub.2PEG.sub.2γEC.sub.18-OH)DFVQWL2αMDT-NH.sub.2  35 TP614 HαMSQGTFTSDYSKYLDARAAK(PEG.sub.2PEG.sub.2γEC.sub.16-OH)-OH)DFVQWL2αMDT-NH.sub.2  36 TP615 HαMSQGTFTSDYSKYLDARAAQK(PEG.sub.2PEG.sub.2γEC.sub.16-OH)FVQWL2αMDT-NH.sub.2  37 TP616 HαMSQGTFTSDYSKYLDARAAK(PEG.sub.2PEG.sub.2γEC.sub.18-OH)DFVQWL2αMDT-NH.sub.2  38 TP617 HαMSQGTFTSDYSKYLDARAAQK(PEG.sub.2PEG.sub.2γEC.sub.18-OH)FVQWL2αMDT-NH.sub.2  39 TP618 HαMSQGTFTSDYSKYLDURAAK(PEG.sub.2PEG.sub.2γEC.sub.16-OH)DFVQWL2αMDT-NH.sub.2  40 TP619 HαMSQGTFTSDYSKYLDURAAQK(PEG.sub.2PEG.sub.2γEC.sub.16-OH)FVQWL2αMDT-NH.sub.2  41 TP620 HαMSQGTFTSDYSKYLDURAAK(PEG.sub.2PEG.sub.2γEC.sub.18-OH)DFVQWL2αMDT-NH.sub.2  42 TP621 HαMSQGTFTSDYSKYLDURAAQK(PEG.sub.2PEG.sub.2γEC.sub.18-OH)FVQWL2αMDT-NH.sub.2  43 TP622 HUQGTFTSDpAF(PEG.sub.2γEC.sub.16-OH)SKYLDARAAQDFVQWL2DT-NH.sub.2  44 TP623 HUQGTFTSDpAF(PEG.sub.2γEC.sub.18-OH)SKYLDARAAQDFVQWL2DT-NH.sub.2  45 TP624 HUQGTFTSDpAF(PEG.sub.2PEG.sub.2γEC.sub.18-OH)SKYLDARAAQDFVQWL2αMDT-NH.sub.2  46 TP625 HUQGTFTSDpAF(PEG.sub.2PEG.sub.2γEC.sub.18-OH)SKYLDARAAQDFVQαMWL2DT-NH.sub.2  47 TP626 HUQGTFTSDYSKYLDARAAK(PEG.sub.2γEC.sub.18-OH)DFVQWL2DT-NH.sub.2  48 TP627 HUQGTFTSDYSKYLDARAAK(PEG.sub.2γEC.sub.16-OH)DFVQWL2αMDT-NH.sub.2  49 TP629 HUQGTFTSDYSKYLDARAAK(PEG.sub.2γEC.sub.20-OH)DFVQWL2αMDT-NH.sub.2  50 TP631 HUQGTFTSDYSKYLDARAAK(PEG.sub.2PEG.sub.2γEC.sub.18-OH)DFVQαMWL2DT-NH.sub.2  51 TP632 HUQGTFTSDYSKYLDARAAK(PEG.sub.2PEG.sub.2γEC.sub.18-OH)DFVQαMWL2αMDT-NH.sub.2  52 TP633 HUQGTFTSDYSKYLDARAAQpAF(PEG.sub.2γEC.sub.18-OH)FVQWLLDT-NH.sub.2  53 TP634 HUQGTFTSDYSKYLDARAAQpAF(PEG.sub.2PEG.sub.2γEC.sub.18-OH)FVQWL2αMDT-NH.sub.2  54 TP635 HUQGTFTSDYSKYLDARAApAF(PEG.sub.2γEC.sub.18-OH)DFVQWLLDT-NH.sub.2  55 TP636 HUQGTFTSDYSKYLDARAApAF(PEG.sub.2PEG2γEC.sub.18-OH)DFVQWL2αMDT-NH.sub.2  56 TP637 HUQGTFTSDYSKYLDARAAQDFVK(PEG.sub.2γEC.sub.18-OH)WL2DT-NH.sub.2  57 TP638 HUQGTFTSDYSKYLDARAAQDFVpAF(PEG.sub.2γEC.sub.18-OH)WLLDT-NH.sub.2  58 TP639 HUQGTFTSDYSKYLDARAAQDFVpAF(γEC.sub.18-OH)WLLDT-NH.sub.2  59 TP657 HUQGTFTSDpAF(PEG.sub.2PEG.sub.2γEC.sub.16-OH)SKYLDURAAQDFVQWLLαMDTKγE-NH.sub.2  60 TP658 HUQGTFTSDpAF(PEG.sub.2PEG.sub.2γEC.sub.16-OH)SKYLDURAAQDFVQWL2ATKγE-NH.sub.2  61 TP659 HsQGTFTSDpAF(PEG2PEG2γEC16OH)SKYLDURAAQDFVQWLLATKγE-NH.sub.2  62 TP660 HαMSQGTFTSDpAF(PEG.sub.2PEG.sub.2γEC.sub.16-OH)SKYLDURAAQDFVQWLLATKγE-NH.sub.2  63 TP661 HαMsQGTFTSDpAF(PEG.sub.2PEG.sub.2γEC.sub.16-OH)SKYLDURAAQDFVQWLLATKγE-NH.sub.2  64 TP662 HUQGTFTSDYSKYLDURAAK(PEG.sub.2PEG.sub.2γEC.sub.16-OH)DFVQWLLATKγE-NH.sub.2  65 TP663 HUQGTFTSDYSKYLDURAApAF(PEG.sub.2PEG.sub.2γEC.sub.16-OH)DFVQWLLATKγE-NH.sub.2  66 TP664 HUQGTFTSDYSKYLDURAAQK(PEG.sub.2PEG.sub.2γEC.sub.16-OH)FVQWLLATKγE-NH.sub.2  67 TP665 HUQGTFTSDYSKYLDURAAQpAF(PEG.sub.2PEG.sub.2γEC.sub.16-OH)FVQWLLATKγE-NH.sub.2  68 TP666 HUQGTFTSDYSKYLDURAAQFVQK(PEG.sub.2PEG.sub.2γEC.sub.16-OH)WLLATKγE-NH.sub.2  69 TP667 HUQGTFTSDYSKYLDURAAQFVQpAF(PEG.sub.2PEG.sub.2γEC.sub.16-OH)WLLATKγE-NH.sub.2  70 TP672 HUQGTFTSDYSKYLDARAAK(PEG.sub.2PEG.sub.2γEC.sub.18-OH)DFVQWL2ET-NH.sub.2  71 TP673 HUQGTFTSDYSKYLDURAAK(PEG.sub.2PEG.sub.2γEC.sub.18-OH)DFVQWL2ET-NH.sub.2  72 TP674 HUQGTFTSDYSKYLEARAAK(PEG.sub.2PEG.sub.2γEC.sub.18-OH)DFVQWL2DT-NH.sub.2  73 TP675 HUQGTFTSDYSKYLDURAAQDFVK(PEG.sub.2PEG.sub.2γEC.sub.18-OH)WL2ET-NH.sub.2  74 TP676 HUQGTFTSDYSKYLDARAAK(PEG.sub.2PEG.sub.2PEG.sub.2γEC.sub.18-OH)DFVQWL2DT-NH.sub.2  75 TP677 HUQGTFTSDYSKYLDARAAK(PEG.sub.2PEG.sub.2PEG.sub.2γEC.sub.18-OH)DFVQWL2αMDT-NH.sub.2  76 TP678 HUQGTFTSDYSKYLDARAApAF(PEG.sub.2PEG.sub.2PEG.sub.2γEC.sub.18-OH)DFVQWLLDT-NH.sub.2  77 TP679 HUQGTFTSDYSKYLDARAAQpAF(PEG.sub.2PEG.sub.2PEG.sub.2γEC.sub.18-OH)FVQWLLDT-NH.sub.2  78 TP680 HUQGTFTSDYSKYLDARAAQDFVK(PEG.sub.2PEG.sub.2PEG.sub.2γEC.sub.18-OH)WL2DT-NH.sub.2  79 TP681 HUQGTFTSDYSKYLDARAAQK(PEG.sub.2PEG.sub.2γEC.sub.18-OH)FVQWL2αMDT-NH.sub.2  80 TP682 HUQGTFTSDYSKYLDURAAQDFVQWLLpAF(PEG.sub.2PEG.sub.2γEC.sub.18-OH)TKγE-NH.sub.2  81 TP683 HUQGTFTSDYSKYLDURAAQDFVQWLLK(PEG.sub.2PEG.sub.2γEC.sub.18-OH)TKγE-NH.sub.2  82 TP685 HsQGTFTSDYSKYLDURAAQDFVQWLLK(PEG.sub.2PEG.sub.2γEC.sub.18-OH)TKγE-NH.sub.2  83 TP693 HsQGTFTSDpAF(γEγEC16)SKYLDARAAQDFVK(PEG2PEG2γEC16-OH)WL2DT-NH.sub.2  84 TP699 HUQGTFTSEYSKKLDARAAQDFVK(PEG.sub.2PEG.sub.2γEC.sub.18-OH)WL2DT-NH.sub.2  85 TP700 HUQGTFTSDYSEYLDKRAAQDFVK(PEG.sub.2PEG.sub.2γEC.sub.18-OH)WL2DT-NH.sub.2  86 TP701 HUQGTFTSDYSKYLDERAAKDFVK(PEG.sub.2PEG.sub.2γEC.sub.18-OH)WL2DT-NH.sub.2  87 TP702 HUQGTFTSDYSKYLDARAAQEFVK(PEG.sub.2PEG.sub.2γEC.sub.18-OH)WL2DT-NH.sub.2  88 TP703 HUQGTFTSEYSKKLDARAAK(PEG.sub.2PEG.sub.2γEC.sub.18-OH)DFVQWL2DT-NH.sub.2  89 TP704 HUQGTFTSDYSKYLDARAAK(PEG.sub.2PEG.sub.2γEC.sub.18-OH)EFVQWL2DT-NH.sub.2  90 TP705 HUQGTFTSDYSKYLDARAAK(PEG.sub.2PEG.sub.2γEC.sub.18-OH)DFVEWL2KT-NH.sub.2  91 TP712 HUQGTFTSDYSKYLDARAANle(1,2,3-triazole-5-PEG.sub.2PEG.sub.2γEC.sub.18-OH)- DFVQWL2DT-NH.sub.2  92 TP713 HUQGTFTSDYSKYLDARAAK(PEG.sub.2PEG.sub.2γEC.sub.18-OH)EFVQWL2αMDT-NH.sub.2  93 TP735 HUQGTFTSDYSKYLDARAAK(PEG.sub.2PEG.sub.2γEC.sub.18-OH)DFVQWL2αMDT-OH  94 TP736 HUQGTFTSDYSKYLDARAAK(PEG.sub.2PEG.sub.2γEC.sub.18-OH)DFVQWL2DT-OH  95 TP737 HUQGTFTSDYSKYLDURAAQDFVK(PEG.sub.2PEG.sub.2γEC.sub.18-OH)WL2DT-OH  96 TP811 HUQGTFTSDYSKYLDURAAQDFVK(PEG.sub.2PEG.sub.2γEC.sub.16-OH)WL2DT-NH.sub.2  97 TP812 HUQGTFTSDYSKYLDARAAK(PEG.sub.2PEG.sub.2γEC.sub.16-OH)EFVQWL2DT-NH.sub.2  98 TP813 HUQGTFTSDYSKYLDARAAQEFVK(PEG.sub.2PEG.sub.2γEC.sub.16-OH)WL2DT-NH.sub.2  99 TP814 HUQGTFTSDYSKYLDARAAK(PEG.sub.2PEG.sub.2γEC.sub.16-OH)DFVQWL2ET-NH.sub.2 100 TP815 HUQGTFTSDYSKYLDURAAQDFVK(PEG.sub.2PEG.sub.2γEC.sub.16-OH WL2ET-NH.sub.2 101 TP825 HUQGTFTSDYSKYLDARAAQK(PEG.sub.2PEG.sub.2γEC.sub.16-OH)FVQWL2ET-NH.sub.2 102 TP826 HUQGTFTSDYSKYLDARAAK(PEG.sub.2PEG.sub.2γEC.sub.16-OH)DFVQWL2γET-NH.sub.2 103 TP827 HUQGTFTSDYSKYLDARAAQK(PEG.sub.2PEG.sub.2γEC.sub.16-OH)FVQWL2γET-NH.sub.2 104 TP828 HUQGTFTSDYSKYLDURAAQDFVK(PEG.sub.2PEG.sub.2γEC.sub.16-OH)WL2γET-NH.sub.2 109 TP564 HUQGTFTSDYSKYLDURAAK(PEG.sub.2PEG.sub.2γEC20-OH)DFVQWL2DT-NH.sub.2 110 TP575 HUQGTFTSDYSKYLDURAAQDFVK(PEG.sub.2PEG.sub.2γEC18-OH)WL2DT-NH.sub.2 111 TP597 HUQGTFTSDYSKLLDARAAK(PEG.sub.2PEG.sub.2γEC18-OH)DFVQWL2DT-NH.sub.2 112 TP598 HUQGTFTSDYSKYLDAKAAK(PEG.sub.2PEG.sub.2γEC18-OH)DFVQWL2DT-NH.sub.2 113 TP604 HUQGTFTSDYSKYLDARAAQDFVQWL2pAF(PEG.sub.2PEG2γEC18-OH)T-NH.sub.2 114 TP443 HUQGTFTSDYSKYLDARAAQpAF(PEG.sub.2PEG.sub.2γEC18-OH)FVQWLLDT-NH.sub.2 115 TP608 HUQGTFTSDYSKYLDARAAQK(PEG.sub.5-γEC18-OH)FVQWL2aMDT-NH.sub.2 116 TP609 HUQGTFTSDYSKYLDARAAQpAF(PEG.sub.2PEG.sub.2γEC18-OH)FVQWLLaMD-T-NH.sub.2 117 TP628 HUQGTFTSDYSKYLDARAAK(PEG.sub.2γEC18-OH)DFVQWL2αMDT-NH.sub.2 118 TP630 HUQGTFTSDYSKYLDARAAK(PEG.sub.2PEG.sub.2γEC16-OH)DFVQαMWL2DT-NH.sub.2 119 TP640 HUQGTFTSDYSKLLDARAAK(PEG.sub.2PEG.sub.2γEC18-OH)DFVQWL2αMDT-NH.sub.2 120 TP829 HsQGTFTSDNle(1,2,3-triazole-4-C.sub.15)SKYLDARAAQDFVQWLLDT-NH.sub.2 121 TP830 HsQGTFTSDNle(1,2,3-triazole-4-γE-C.sub.16)SKYLDARAAQDFVQWLLDT-NH.sub.2 122 TP831 HsQGTFTSDNle(1,2,3-triazole-4-C.sub.4-Lys-C.sub.16)SKYLDARAAQDFVQWLLDT- NH.sub.2 Table legend: U = alpha-aminoisobutyric acid; γE = γ-glutamic acid; 2 = L-methionine sulphone; αMD = alpha-Methyl-L-Aspartic acid; Nle = norleucine; αMF = alpha-Methyl-L-phenylalanine; αMS = alpha-Methyl-L-serine; αMW = alpha-methyl-L-tryptophan; s = D-serine; pAF = p-aminomethyl-L- phenylalanine; PEG.sub.2 = 8-amino-3,6-dioxaoctanoic acid; PEG.sub.5 = 1-hydroxy-3,6,9,12,15-pentaoxaoctadecan-18-oic acid; C.sub.4 is C.sub.4 alkyl; C.sub.15 = C.sub.15 alkyl; C.sub.16 = C.sub.16 alkyl; C.sub.x = C.sub.x alkyl; C.sub.16—OH or C16—OH = —CO—(CH.sub.2).sub.14—COOH; C.sub.18—OH or C18—OH = —CO—(CH.sub.2).sub.16—COOH; C.sub.20—OH or C20—OH = —CO—(CH.sub.2).sub.18—COOH; NH.sub.2 = C-terminal amide..

(78) The structure of Nle(1,2,3-triazole-5-PEG.sub.2PEG.sub.2γEC.sub.18—OH) wherein the linker is PEG.sub.2PEG.sub.2γE and the fatty acid component comprises C18-OH is represented by (peptide SEQ ID NO:91):

(79) ##STR00001##

(80) The GCG/GLP-1 receptor co-agonist peptides of the present invention are conjugated to an α,ω-dicarboxylic acid comprising an aliphatic chain of 14 to 20 methylene groups (fatty diacid) wherein one end of the molecule is the proximal end and the other end is the distal end and wherein the proximal end and the distal end both have a carboxyl (COOH) group. The fatty diacid may be represented by the structure HO.sub.2C(CH.sub.2).sub.nCO.sub.2H, wherein n is 11, 12, 13, 14, 15, 16, 17 or 18. The fatty diacids include but are not limited to, the fatty diacids Tetradecanedioic acid, Hexadecanedioic acid, Heptadecanedioic acid, Octadecanedioic acid, Nonadecanedioic acid, and Eicosanedioic acid, respectively. The aforementioned fatty diacids have the following structures

(81) ##STR00002##

(82) In particular aspects, the GCG/GLP-1 receptor co-agonist peptide is further conjugated to a fatty acid at position 10 of the peptide. The fatty acid may be represented by the structure HO.sub.2C(CH.sub.2).sub.n wherein n is 11, 12, 13, 14, 15, 16, 17, 18, 19 or 20. The fatty acid may have one of the following structures

(83) ##STR00003##

(84) As a component of the GCG/GLP-1 receptor co-agonist peptide, the acid functionality at the proximal end of the fatty diacid is conjugated to the amino group of a linker in a C(O)—NH linkage and the acid functionality at the distal end of the fatty diacid is a free carboxyl group (COOH). The COOH group at the distal end helps confer a longer half-life to the co-agonist peptide by its ability to non-covalently bind to serum albumin, a known carrier for fatty acids in serum. The COOH group enhances duration of action as it provides a better non-covalent interaction with serum albumin than GCG/GLP-1 receptor co-agonist peptides that have been acylated using a fatty acid, which bind serum albumin less efficiently and form a less stable non-covalent interaction with the serum albumin. When the fatty diacid is conjugated to a linking moiety, it is subsequently referred to as a fatty acid component.

(85) The linker may be PEG.sub.2 (8-amino-3,6-dioxaoctanoic acid) linked to Gamma-Glutamic acid (gamma-Glu, γGlu, or γE), which has the structure

(86) ##STR00004##
or the linker may be Gamma-Glutamic acid-gamma glutamic acid (gamma-Glu-gamma-Glu, or γGlu-γGlu, or γEγE), which has the structure

(87) ##STR00005##

(88) The linker may also be Peg.sub.5(1-hydroxy-3,6,9,12,15-pentaoxaoctadecan-18-oic acid) linked to Gamma-Glutamic acid (gamma-Glu, γGlu, or γE), which has the structure

(89) ##STR00006##

(90) The structure of K(PEG.sub.2PEG.sub.2γE-fatty acid) wherein the linker is PEG.sub.2PEG.sub.2γE and the fatty acid component comprises C14, C16, C17, C18, C19, or C20 is represented by

(91) ##STR00007##
wherein n is 7, 9, 10, 11, 12, 13, or 14 respectively, and the wavy lines represent the bonds between adjacent amino acids in the co-agonist peptide sequence.

(92) The structure of pAF(PEG.sub.2PEG.sub.2γE-fatty acid) wherein the linker is PEG.sub.2PEG.sub.2γE and the fatty acid component comprises C14, C16, C17, C18, C19, or C20 is represented by

(93) ##STR00008##
wherein n is 7, 9, 10, 11, 12, 13, or 14, respectively, and the wavy lines represent the bonds between adjacent amino acids in the co-agonist peptide sequence.

(94) The structure of K(γEγE-fatty acid) wherein the linker is γEγE and the fatty acid component comprises C14, C16, C17, C18, C19, or C20 is represented by

(95) ##STR00009##
wherein n is 7, 9, 10, 11, 12, 13, or 14, respectively, and the wavy lines represent the bonds between adjacent amino acids in the co-agonist peptide sequence.

(96) The structure of KγE at position 30 in the co-agonist peptide is represented by

(97) ##STR00010##
wherein the wavy line represents the bond between adjacent amino acids in the co-agonist peptide sequence, and wherein X is OH or NH.sub.2.

(98) The GCG/GLP-1 receptor co-agonist peptides disclosed herein may have anywhere from at least about 1% (including at least about 1.5%, 2%, 5%, 7%, 10%, 20%, 30%, 40%, 50%, 60%, 75%, 100%, 125%, 150%, 175%) to about 200% or higher activity at the GLP-1 receptor relative to native GLP-1 and anywhere from at least about 1% (including about 1.5%, 2%, 5%, 7%, 10%, 20%, 30%, 40%, 50%, 60%, 75%, 100%, 125%, 150%, 175%, 200%, 250%, 300%, 350%, 400%, 450%) to about 500% or higher activity at the glucagon receptor relative to native glucagon.

(99) In some embodiments, the GCG/GLP-1 receptor co-agonist peptides described herein exhibit no more than about 100%, 1000%, 10,000%, 100,000%, or 1,000,000% of the activity of native glucagon at the glucagon receptor.

(100) In some embodiments, the GCG/GLP-1 receptor co-agonist peptides described herein exhibit no more than about 100%, 1000%, 10,000%, 100,000%, or 1,000,000% of the activity of native GLP-1 at the GLP-1 receptor.

(101) In exemplary embodiments, a GCG/GLP-1 receptor co-agonist peptide may exhibit at least 10% of the activity of native glucagon at the glucagon receptor and at least 50% of the activity of native GLP-1 at the GLP-1 receptor, or at least 40% of the activity of native glucagon at the glucagon receptor and at least 40% of the activity of native GLP-1 at the GLP-1 receptor, or at least 60% of the activity of native glucagon at the glucagon receptor and at least 60% of the activity of native GLP-1 at the GLP-1 receptor.

(102) Selectivity of a GCG/GLP-1 receptor peptide for the glucagon receptor versus the GLP-1 receptor can be described as the relative ratio of glucagon/GLP-1 activity (the peptide analog's activity at the glucagon receptor relative to native glucagon, divided by the peptide's activity at the GLP-1 receptor relative to native GLP-1). For example, a GCG/GLP-1 receptor co-agonist peptide that exhibits 60% of the activity of native glucagon at the glucagon receptor and 60% of the activity of native GLP-1 at the GLP-1 receptor has a 1:1 ratio of glucagon/GLP-1 activity. Exemplary ratios of glucagon/GLP-1 activity include about 1:1, 1.5:1, 2:1, 3:1, 4:1, 5:1, 6:1, 7:1, 8:1, 9:1 or 10:1, or about 1:10, 1:9, 1:8, 1:7, 1:6, 1:5, 1:4, 1:3, 1:2, or 1:1.5. As an example, a glucagon/GLP-1 activity ratio of 10:1 indicates a 10-fold selectivity for the glucagon receptor versus the GLP-1 receptor. Similarly, a GLP-1/glucagon activity ratio of 10:1 indicates a 10-fold selectivity for the GLP-1 receptor versus the glucagon receptor.

Pharmaceutical Compositions

(103) Further provided are pharmaceutical compositions comprising a therapeutically effective amount of one or more of the GCG/GLP-1 receptor co-agonist peptides disclosed herein for the treatment of a metabolic disorder in an individual. Such disorders include, but are not limited to, obesity, metabolic syndrome or syndrome X, Type II diabetes, complications of Type II diabetes such as retinopathy, hypertension, dyslipidemias, cardiovascular disease, gallstones, osteoarthritis, and certain forms of cancers. The obesity-related disorders herein are associated with, caused by, or result from obesity.

(104) “Obesity” is a condition in which there is an excess of body fat. The operational definition of obesity is based on the Body Mass Index (BMI), calculated as body weight per height in meters squared (kg/m2). “Obesity” refers to a condition whereby an otherwise healthy subject has a Body Mass Index (BMI) greater than or equal to 30 kg/m2, or a condition whereby a subject with at least one co-morbidity has a BMI greater than or equal to 27 kg/m2. An “obese subject” is an otherwise healthy subject with a Body Mass Index (BMI) greater than or equal to 30 kg/m2 or a subject with at least one co-morbidity with a BMI greater than or equal to 27 kg/m2. A “subject at risk for obesity” is an otherwise healthy subject with a BMI of 25 kg/m2 to less than 30 kg/m2 or a subject with at least one co-morbidity with a BMI of 25 kg/m2 to less than 27 kg/m2.

(105) The increased risks associated with obesity occur at a lower Body Mass Index (BMI) in Asians. In Asian countries, including Japan, “obesity” refers to a condition whereby a subject with at least one obesity-induced or obesity-related co-morbidity that requires weight reduction or that would be improved by weight reduction, has a BMI greater than or equal to 25 kg/m2. In Asian countries, including Japan, an “obese subject” refers to a subject with at least one obesity-induced or obesity-related co-morbidity that requires weight reduction or that would be improved by weight reduction, with a BMI greater than or equal to 25 kg/m2. In Asian countries, a “subject at risk of obesity” is a subject with a BMI of greater than 23 kg/m2 to less than 25 kg/m2.

(106) As used herein, the term “obesity” is meant to encompass all of the above definitions of obesity.

(107) Obesity-induced or obesity-related co-morbidities include, but are not limited to, Type II diabetes, impaired glucose tolerance, impaired fasting glucose, insulin resistance syndrome, dyslipidemia, hypertension, hyperuricacidemia, gout, coronary artery disease, myocardial infarction, angina pectoris, sleep apnea syndrome, Pickwickian syndrome, non-alcoholic fatty liver disease (NAFLD), non-alcoholic steatohepatitis (NASH), fatty liver; cerebral infarction, cerebral thrombosis, transient ischemic attack, orthopedic disorders, arthritis deformans, lumbodynia, emmeniopathy, and infertility. In particular, co-morbidities include: hypertension, hyperlipidemia, dyslipidemia, glucose intolerance, cardiovascular disease, sleep apnea, diabetes mellitus, and other obesity-related conditions.

(108) “Treatment” (of obesity and obesity-related disorders) refers to the administration of a GCG/GLP-1 receptor co-agonist peptide to reduce or maintain the body weight of an obese subject. One outcome of treatment may be reducing the body weight of an obese subject relative to that subject's body weight immediately before the administration of the compounds of the present invention. Another outcome of treatment may be preventing body weight regain of body weight previously lost as a result of diet, exercise, or pharmacotherapy. Another outcome of treatment may be decreasing the occurrence of and/or the severity of obesity-related diseases. The treatment may suitably result in a reduction in food or calorie intake by the subject, including a reduction in total food intake, or a reduction of intake of specific components of the diet such as carbohydrates or fats; and/or the inhibition of nutrient absorption; and/or the inhibition of the reduction of metabolic rate; and in weight reduction in patients in need thereof. The treatment may also result in an alteration of metabolic rate, such as an increase in metabolic rate, rather than or in addition to an inhibition of the reduction of metabolic rate; and/or in minimization of the metabolic resistance that normally results from weight loss.

(109) “Prevention” (of obesity and obesity-related disorders) refers to the administration of the compounds of the present invention to reduce or maintain the body weight of a subject at risk of obesity. One outcome of prevention may be reducing the body weight of a subject at risk of obesity relative to that subject's body weight immediately before the administration of the compounds of the present invention. Another outcome of prevention may be preventing body weight regain of body weight previously lost as a result of diet, exercise, or pharmacotherapy. Another outcome of prevention may be preventing obesity from occurring if the treatment is administered prior to the onset of obesity in a subject at risk of obesity. Another outcome of prevention may be decreasing the occurrence and/or severity of obesity-related disorders if the treatment is administered prior to the onset of obesity in a subject at risk of obesity. Moreover, if treatment is commenced in already obese subjects, such treatment may prevent the occurrence, progression or severity of obesity-related disorders, such as, but not limited to, arteriosclerosis, Type II diabetes, polycystic ovarian disease, cardiovascular diseases, osteoarthritis, dermatological disorders, hypertension, insulin resistance, hypercholesterolemia, hypertriglyceridemia, and cholelithiasis.

(110) The obesity-related disorders herein are associated with, caused by, or result from obesity. Examples of obesity-related disorders include overeating and bulimia, hypertension, diabetes, elevated plasma insulin concentrations and insulin resistance, dyslipidemias, hyperlipidemia, endometrial, breast, prostate and colon cancer, osteoarthritis, obstructive sleep apnea, cholelithiasis, gallstones, heart disease, abnormal heart rhythms and arrythmias, myocardial infarction, congestive heart failure, coronary heart disease, sudden death, stroke, polycystic ovarian disease, craniopharyngioma, the Prader-Willi Syndrome, Frohlich's syndrome, GH-deficient subjects, normal variant short stature, Turner's syndrome, and other pathological conditions showing reduced metabolic activity or a decrease in resting energy expenditure as a percentage of total fat-free mass, e.g, children with acute lymphoblastic leukemia. Further examples of obesity-related disorders are metabolic syndrome, also known as syndrome X, insulin resistance syndrome, sexual and reproductive dysfunction, such as infertility, hypogonadism in males and hirsutism in females, gastrointestinal motility disorders, such as obesity-related gastro-esophageal reflux, respiratory disorders, such as obesity-hypoventilation syndrome (Pickwickian syndrome), cardiovascular disorders, inflammation, such as systemic inflammation of the vasculature, arteriosclerosis, hypercholesterolemia, hyperuricaemia, lower back pain, gallbladder disease, gout, and kidney cancer. The compounds of the present invention are also useful for reducing the risk of secondary outcomes of obesity, such as reducing the risk of left ventricular hypertrophy.

(111) The term “diabetes,” as used herein, includes both insulin-dependent diabetes mellitus (IDDM, also known as type I diabetes) and non-insulin-dependent diabetes mellitus (NIDDM, also known as Type II diabetes). Type I diabetes, or insulin-dependent diabetes, is the result of an absolute deficiency of insulin, the hormone which regulates glucose utilization. Type II diabetes, or insulin-independent diabetes (i.e., non-insulin-dependent diabetes mellitus), often occurs in the face of normal, or even elevated levels of insulin and appears to be the result of the inability of tissues to respond appropriately to insulin. Most of the Type II diabetics are also obese. The GCG/GLP-1 receptor co-agonist peptides are useful for treating both Type I and Type II diabetes. The GCG/GLP-1 receptor co-agonist peptides are especially effective for treating Type II diabetes. The GCG/GLP-1 receptor co-agonist peptides are also useful for treating and/or preventing gestational diabetes mellitus.

(112) U.S. Pat. No. 6,852,690, which is incorporated herein in its entirety, discloses methods for enhancing metabolism of nutrients comprising administering to a non-diabetic patient a formulation comprising a nutritively effective amount of one or more nutrients or any combination thereof and one or more insulinotropic peptides. The GCG/GLP-1 receptor co-agonist peptides disclosed herein are insulinotropic and can be administered to patients with a disturbed glucose metabolism such as insulin resistance but no overt diabetes, as well as patients who for any reason cannot receive nutrition through the alimentary canal. Such patients include surgery patients, comatose patients, patients in shock, patients with gastrointestinal disease, patients with digestive hormone disease, and the like. In particular, obese patients, atherosclerotic patients, vascular disease patients, patients with gestational diabetes, patients with liver disease such as liver cirrhosis, patients with acromegaly, patients with glucorticoid excess such as cortisol treatment or Cushings disease, patients with activated counterregulatory hormones such as would occur after trauma, accidents and surgery and the like, patients with hypertriglyceridemia and patients with chronic pancreatitis can be readily and suitably nourished according to the invention without subjecting the patient to hypo- or hyperglycemia. In particular, the administration to such a patient aims to provide a therapy to as rapidly as possible deliver the nutritional and caloric requirements to the patient while maintaining his plasma glucose below the so-called renal threshold of about 160 to 180 milligrams per deciliter of glucose in the blood. Although normal patients not having glucose levels just below the renal threshold can also be treated according to the invention as described above, patients with disturbed glucose metabolism such as hyperglycemic patients whose plasma glucose level is just above the renal threshold also find the therapy suitable for their condition. In particular, such patients who have a degree of hyperglycemia below the renal threshold at intermittent intervals can receive a combination treatment of nutrients plus insulinotropic peptides according to any of the following regimens. Normal patients not suffering from such hyperglycemia can also be treated using the GCG/GLP-1 receptor co-agonist peptides disclosed herein.

(113) The GCG/GLP-1 receptor co-agonist peptides disclosed herein may be used in a pharmaceutical composition when combined with a pharmaceutically acceptable carrier. Such compositions comprise a therapeutically-effective amount of one or more of the co-agonist peptides disclosed herein and a pharmaceutically acceptable carrier. Such a composition may also be comprised of (in addition to the co-agonist peptides disclosed herein and a carrier) diluents, fillers, salts, buffers, stabilizers, solubilizers, and other materials well known in the art. Compositions comprising the co-agonist peptides disclosed herein can be administered, if desired, in the form of salts provided the salts are pharmaceutically acceptable. Salts may be prepared using standard procedures known to those skilled in the art of synthetic organic chemistry.

(114) The term “individual” is meant to include humans and companion or domesticated animals such as dogs, cats, horses, and the like. Therefore, the compositions comprising a compound as disclosed herein are also useful for treating or preventing obesity and obesity-related disorders in cats and dogs. As such, the term “mammal” includes companion animals such as cats and dogs.

(115) The term “pharmaceutically acceptable salts” refers to salts prepared from pharmaceutically acceptable non-toxic bases or acids including inorganic or organic bases and inorganic or organic acids. Salts derived from inorganic bases include aluminum, ammonium, calcium, copper, ferric, ferrous, lithium, magnesium, manganic salts, manganous, potassium, sodium, zinc, and the like. Particularly preferred are the ammonium, calcium, magnesium, potassium, and sodium salts. Salts derived from pharmaceutically acceptable organic non-toxic bases include salts of primary, secondary, and tertiary amines, substituted amines including naturally occurring substituted amines, cyclic amines, and basic ion exchange resins, such as arginine, betaine, caffeine, choline, N,N′-dibenzylethylenediamine, diethylamine, 2-diethylaminoethanol, 2-dimethylaminoethanol, ethanolamine, ethylenediamine, N-ethyl-morpholine, N-ethylpiperidine, glucamine, glucosamine, histidine, hydrabamine, isopropylamine, lysine, methylglucamine, morpholine, piperazine, piperidine, polyamine resins, procaine, purines, theobromine, triethylamine, trimethylamine, tripropylamine, tromethamine, and the like. The term “pharmaceutically acceptable salt” further includes all acceptable salts such as acetate, lactobionate, benzenesulfonate, laurate, benzoate, malate, bicarbonate, maleate, bisulfate, mandelate, bitartrate, mesylate, borate, methylbromide, bromide, methylnitrate, calcium edetate, methylsulfate, camsylate, mucate, carbonate, napsylate, chloride, nitrate, clavulanate, N-methylglucamine, citrate, ammonium salt, dihydrochloride, oleate, edetate, oxalate, edisylate, pamoate (embonate), estolate, palmitate, esylate, pantothenate, fumarate, phosphate/diphosphate, gluceptate, polygalacturonate, gluconate, salicylate, glutamate, stearate, glycollylarsanilate, sulfate, hexylresorcinate, subacetate, hydrabamine, succinate, hydrobromide, tannate, hydrochloride, tartrate, hydroxynaphthoate, teoclate, iodide, tosylate, trifluoro acetate, isothionate, triethiodide, lactate, panoate, valerate, and the like which can be used as a dosage form for modifying the solubility or hydrolysis characteristics or can be used in sustained release or pro-drug formulations. It will be understood that, as used herein, references to the GCG/GLP-1 receptor co-agonist peptides disclosed herein are meant to also include the pharmaceutically acceptable salts. The invention also includes counterions, including pharmaceutically acceptable counterions, including but not limited to, sodium, acetate and trifluoro acetate.

(116) As utilized herein, the term “pharmaceutically acceptable” means a non-toxic material that does not interfere with the effectiveness of the biological activity of the active ingredient(s), approved by a regulatory agency of the Federal or a state government or listed in the U.S. Pharmacopoeia or other generally recognized pharmacopoeia for use in animals and, more particularly, in humans. The term “carrier” refers to a diluent, adjuvant, excipient, or vehicle with which the therapeutic is administered and includes, but is not limited to such sterile liquids as water and oils. The characteristics of the carrier will depend on the route of administration. The GCG/GLP-1 receptor co-agonist peptides disclosed herein may be in multimers (for example, heterodimers or homodimers) or complexes with itself or other peptides. As a result, pharmaceutical compositions of the invention may comprise one or more co-agonist peptides disclosed herein in such multimeric or complexed form.

(117) As used herein, the term “therapeutically effective amount” means the total amount of each active component of the pharmaceutical composition or method that is sufficient to show a meaningful patient benefit, i.e., treatment, healing, prevention or amelioration of the relevant medical condition, or an increase in rate of treatment, healing, prevention or amelioration of such conditions. When applied to an individual active ingredient, administered alone, the term refers to that ingredient alone. When applied to a combination, the term refers to combined amounts of the active ingredients that result in the therapeutic effect, whether administered in combination, serially, or simultaneously.

(118) The pharmacological composition may comprise one or more co-agonist peptides disclosed herein; one or more co-agonist peptides disclosed herein and one or more other agents for treating a metabolic disorder; or the pharmacological composition comprising the one or more co-agonist peptides disclosed herein can be used concurrently with a pharmacological composition comprising an agent for treating a metabolic disorder. Such disorders include, but are not limited to, obesity, metabolic syndrome or syndrome X, type II diabetes, complications of diabetes, hypertension, dyslipidemias, cardiovascular disease, gallstones, osteoarthritis, and certain forms of cancers.

(119) When a GCG/GLP-1 receptor co-agonist peptide is used contemporaneously with one or more other drugs, peptides, or proteins, a pharmaceutical composition containing such other drugs, peptides, or proteins in addition to the GCG/GLP-1 receptor co-agonist peptide may be provided. Accordingly, the pharmaceutical compositions of the present invention include those that also contain one or more other active ingredients, in addition to a GCG/GLP-1 receptor co-agonist peptide. Examples of other proteins that may be included in the composition include but are not limited to human insulin or human insulin analog such as insulin detemir, insulin glargine (U100 or U300), insulin levemir, insulin glulisine, insulin degludec, or insulin lispro.

(120) Methods of administrating the pharmacological compositions comprising the one or more GCG/GLP-1 receptor co-agonist peptides disclosed herein to an individual include, but are not limited to, intradermal, intramuscular, intraperitoneal, intravenous, subcutaneous, intranasal, epidural, and oral routes. The compositions can be administered by any convenient route, for example by infusion or bolus injection, by absorption through epithelial or mucocutaneous linings (for example, oral mucosa, rectal and intestinal mucosa, and the like), ocular, and the like and can be administered together with other biologically-active agents. Administration can be systemic or local.

(121) Various delivery systems are known and can be used to administer the GCG/GLP-1 receptor co-agonist peptides disclosed herein including, but not limited to, encapsulation in liposomes, microparticles, microcapsules; minicells; polymers; capsules; tablets; and the like. In one embodiment, the co-agonist peptides disclosed herein may be delivered in a vesicle, in particular a liposome. In a liposome, the co-agonist peptides disclosed herein are combined, in addition to other pharmaceutically acceptable carriers, with amphipathic agents such as lipids which exist in aggregated form as micelles, insoluble monolayers, liquid crystals, or lamellar layers in aqueous solution. Suitable lipids for liposomal formulation include, without limitation, monoglycerides, diglycerides, sulfatides, lysolecithin, phospholipids, saponin, bile acids, and the like. Preparation of such liposomal formulations is within the level of skill in the art, as disclosed, for example, in U.S. Pat. Nos. 4,837,028 and 4,737,323. In yet another embodiment, the co-agonist peptides disclosed herein can be delivered in a controlled release system including, but not limited to: a delivery pump (See, for example, Saudek, et al., New Engl. J. Med. 321: 574 (1989) and a semi-permeable polymeric material (See, for example, Howard, et al., J. Neurosurg. 71: 105 (1989)). Additionally, the controlled release system can be placed in proximity of the therapeutic target (for example, the brain), thus requiring only a fraction of the systemic dose. See, for example, Goodson, In: Medical Applications of Controlled Release, 1984. (CRC Press, Bocca Raton, Fla.).

(122) The amount of the compositions comprising one or more of the GCG/GLP-1 receptor co-agonist peptides disclosed herein which will be effective in the treatment of a particular disorder or condition will depend on the nature of the disorder or condition, and may be determined by standard clinical techniques by those of average skill within the art. In addition, in vitro assays may optionally be employed to help identify optimal dosage ranges. The precise dose to be employed in the formulation will also depend on the route of administration, and the overall seriousness of the disease or disorder, and should be decided according to the judgment of the practitioner and each patient's circumstances. Ultimately, the attending physician will decide the amount of the composition with which to treat each individual patient. Initially, the attending physician will administer low doses of the composition and observe the patient's response. Larger doses of the composition may be administered until the optimal therapeutic effect is obtained for the patient, and at that point the dosage is not increased further. In general, the daily dose range lie within the range of from about 0.001 mg to about 100 mg per kg body weight of a mammal, preferably 0.01 mg to about 50 mg per kg, and most preferably 0.1 to 10 mg per kg, in single or divided doses. On the other hand, it may be necessary to use dosages outside these limits in some cases. However, suitable dosage ranges for intravenous administration of the compositions comprising the one or more co-agonist peptides disclosed herein are generally about 5-500 micrograms (μg) of active compound per kilogram (Kg) body weight. Suitable dosage ranges for intranasal administration are generally about 0.01 pg/kg body weight to 1 mg/kg body weight. Effective doses may be extrapolated from dose-response curves derived from in vitro or animal model test systems. Suppositories generally contain active ingredient in the range of 0.5% to 10% by weight; oral formulations preferably contain 10% to 95% active ingredient. The peptides may be administered on a regimen including, but not limited to, 1 to 4 times per day, once every 2 days, once every 3 days, once every 4 days, once every 5 days once every 6 days or once a week. Ultimately the attending physician will decide on the appropriate duration of therapy using compositions comprising the one or more co-agonist peptides disclosed herein of the present invention. Dosage will also vary according to the age, weight and response of the individual patient.

(123) Further provided is a pharmaceutical pack or kit, comprising one or more containers filled with one or more of the ingredients of the pharmaceutical compositions and co-agonist peptides disclosed herein. Optionally associated with such container(s) may be a notice in the form prescribed by a governmental agency regulating the manufacture, use or sale of pharmaceuticals or biological products, which notice reflects approval by the agency of manufacture, use or sale for human administration.

(124) The present invention further provides a method for treating a patient for a metabolic disease or disorder comprising administering the patient an effective amount of a composition comprising any one or more of the aforementioned GCG/GLP-1 receptor co-agonist peptides to treat the metabolic disease or disorder in the patient.

(125) In particular aspects, the metabolic disease or disorder is diabetes, non-alcoholic fatty liver disease (NAFLD), non-alcoholic steatohepatitis (NASH), or obesity. In particular aspects, the diabetes is Type I diabetes, Type II diabetes, or gestational diabetes. In particular aspects, the patient has more than one metabolic disease or disorder, for example, diabetes and NASH, NAFLD, or obesity; obesity and NASH or NAFLD; diabetes, NASH, and obesity; diabetes, NAFLD, and obesity; or diabetes and obesity.

(126) The present invention further provides for the use of any one of the aforementioned GCG/GLP-1 receptor co-agonist peptides for manufacture of a medicament for the treatment of a metabolic disease or disorder.

(127) The present invention further provides for the use of any one of the aforementioned GCG/GLP-1 receptor co-agonist peptides for manufacture of a medicament for the treatment of a metabolic disease or disorder.

(128) In particular aspects, the metabolic disease or disorder is diabetes, non-alcoholic fatty liver disease (NAFLD), non-alcoholic steatohepatitis (NASH), or obesity. In particular aspects, the diabetes is Type I diabetes, Type II diabetes, or gestational diabetes. In particular aspects, the medicament is for treatment of more than one metabolic disease or disorder, for example, diabetes and NASH, NAFLD, or obesity; obesity and NASH or NAFLD; diabetes, NASH, and obesity; diabetes, NAFLD, and obesity; or diabetes and obesity.

(129) Further provided is method for treating a metabolic disease or disorder in a patient or individual comprising: administering to the patient or individual an effective amount of any one of the aforementioned compositions comprising a GCG/GLP-1 receptor co-agonist peptide and administering to the patient or individual an effective amount of a composition comprising an insulin or insulin analog to treat the metabolic disease or disorder in the patient or individual.

(130) In particular aspects, the composition comprising the GCG/GLP-1 receptor co-agonist peptide is administered at a time prior to the time the composition comprising the insulin or insulin analog is administered. In another aspect, the composition comprising the insulin or insulin analog is administered at a time prior to the time the composition comprising the co-agonist peptide is administered. In a further still aspect, the composition comprising the co-agonist peptide is administered at the same time as the composition comprising the insulin or insulin analog is administered.

(131) In particular aspects, the insulin is human insulin or a human insulin analog such as insulin detemir, insulin glargine (U100 or U300), insulin levemir, insulin glulisine, insulin degludec, or insulin lispro.

(132) In particular aspects, the metabolic disease or disorder is diabetes, non-alcoholic fatty liver disease (NAFLD), non-alcoholic steatohepatitis (NASH), or obesity. In particular aspects, the diabetes is Type I diabetes, Type II diabetes, or gestational diabetes. In particular aspects, the patient has more than one metabolic disease or disorder, for example, diabetes and NASH, NAFLD, or obesity; obesity and NASH or NAFLD; diabetes, NASH, and obesity; diabetes, NAFLD, and obesity; or diabetes and obesity.

(133) The present invention further provides a composition comprising any one of the aforementioned GCG/GLP-1 receptor co-agonist peptides; an insulin or insulin analog; and, a pharmaceutically acceptable carrier.

(134) The present invention further provides for the use of a composition comprising any one of the aforementioned GCG/GLP-1 receptor co-agonist peptides; an insulin or insulin analog; and, a pharmaceutically acceptable carrier for the treatment of a metabolic disease or disorder.

(135) In particular aspects, the metabolic disease or disorder is diabetes, non-alcoholic fatty liver disease (NAFLD), non-alcoholic steatohepatitis (NASH), or obesity. In further aspects, the diabetes is Type I diabetes, Type II diabetes, or gestational diabetes.

(136) The present invention further provides for the use of a composition comprising any one of the aforementioned GCG/GLP-1 receptor co-agonist peptides; an insulin or insulin analog; and, a pharmaceutically acceptable carrier for the manufacture of a medicament for the treatment of a metabolic disease or disorder.

(137) In particular aspects, the insulin analog comprises insulin detemir, insulin glargine (U100 or U300), insulin levemir, insulin glulisine, insulin degludec, or insulin lispro.

(138) In particular aspects, the metabolic disease or disorder is diabetes, non-alcoholic fatty liver disease (NAFLD), non-alcoholic steatohepatitis (NASH), or obesity. In further aspects, the diabetes is Type I diabetes, Type II diabetes, or gestational diabetes.

(139) The following examples are intended to promote a further understanding of the present invention.

EXAMPLE 1

(140) Peptides in Table 1 were synthesized by solid phase synthesis using Fmoc/t-Bu chemistry on a peptide multisynthesizer Symphony (Protein Technologies Inc.) on a 150 μmol scale, using either a Rink-amide PEG-PS resin (Champion, Biosearch Technologies, loading 0.28 mmol/g) or a Rink-amide PS resin (ChemImpex loading 0.47 mmol/g).

(141) All the amino acids were dissolved at a 0.3 M concentration in DMF. The amino acids were activated with equimolar amounts of HATU (O-(7-azabenzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium hexafluorophosphate) solution 0.3 M in DMF, and a 2-fold molar excess of DIEA (N,N-diisopropylethylamine), solution 2M in NMP. The acylation reactions were performed in general for 1 hour with a 5-fold excess of activated amino acid over the resin free amino groups with double 45 minutes acylation reactions performed from His.sup.1 to Thr.sup.7 and from F.sup.22 to V.sup.23 and from D/E.sup.15 to Aib.sup.16 for sequences containing Aib.sup.16.

(142) The side chain protecting groups were: tert-butyl for Asp, αMD, Glu, Ser, αMS, D-Ser, Thr and Tyr; trityl for Gln and His; tert-butoxy-carbonyl for Lys, Trp; and, 2,2,4,6,7-pentamethyldihydrobenzofuran-5-sulfonyl for Arg; His was introduced as Boc-His(Trt)—OH at the end of the sequence assembly. Amino acid 2 (L-methionine-sulphone) was introduced by acylation of Fmoc-L-methionine-sulphone-COOH. For all the sequences containing alpha methyl amino acids, incorporation of the alpha methyl amino acid and the corresponding following residue were performed by manual coupling with HOAt (Hydroxybenzoazatriazole) and DIC (N,N′-diisopropylcarbodiimide). The position used for linker-lipid derivatization, either lysine or pAF (p-aminomethyl-L-phenylalanine) were incorporated with a Dde [1-(4,4-dimethyl-2,6-dioxocyclohex-1-ylidene)ethyl] or Alloc (Allyloxy-carbonyl) protecting groups on the side chain amino group. In sequence ID 91 the position used for linker derivativation was incorporated as Fmoc-Nle(1,2,3-triazole-5-PEG.sub.2-NH-ivDde as manual coupling using HOAt, DIC. For sequences with double lipids, sequences ID 25, 29 and 30, position 10 was incorporated as Lys(Alloc)—OH and the second position for the lipid diacid derivatization was incorporated as Lys(Dde)—OH. The pAF [Fmoc-4-(Dde-aminomethyl)-phenylalanine] and the Fmoc-Nle(1,2,3-triazole-5-PEG.sub.2-NH-ivDde amino acids were synthesized as described below.

(143) At the end of the sequence assembly, to proceed with linker/lipid derivatization, the Dde protecting group of either pAF(Dde) or Lys(Dde) or Nle(1,2,3-triazole-5-PEG.sub.2-NH-ivDde was removed by treatment of 2% hydrazine in DMF. The side chains of Lys or pAF or Nle(1,2,3-triazole-5-PEG.sub.2-NH.sub.2 were derivatized with different linkers and fatty diacids by incorporation of Fmoc-Glu-OtBu (γ-glutamic acid), Fmoc-PEG2 [8-(9-Fluorenylmethyloxycarbonyl-amino)-3,6-dioxaoctanoic acid] and the lipid diacids (; Hexadecanedioic acid; Octadecanedioic acid; Eicosanedioic acid) using HOAt and DIC as activators.

(144) For double lipidated sequences ID 25, 29, 30, the Alloc protecting group from Lys10 was first removed by treatment with Pd(PPh3)4 and PhSiH3 in DCM (dichloromethane) followed by derivatization with Fmoc-Glu-OtBu (γ-glutamic acid) and hexanenoic acid. Then deprotection of the Dde from the other Lys followed by derivatization with linker/lipid diacids was performed as described above for the other lapidated analogs.

(145) At the end of the synthesis, the dry peptide-resins were individually treated with 25 mL of the cleavage mixture, 88% TFA, 5% phenol, 2% triisopropylsilane and 5% water for 1.5 hours at room temperature. Each resin was filtered and then added to cold methyl-t-butyl ether in order to precipitate the peptide. After centrifugation, the peptide pellets were washed with fresh cold methyl-t-butyl ether to remove the organic scavengers. The process was repeated twice. Final pellets were dried, resuspended in H.sub.2O, 20% acetonitrile, and lyophilized. The crude peptides (140 mg in 3 mL of DMSO) were purified by reverse-phase HPLC using preparative Waters Deltapak C4 (40×200 mm, 15 μm, 300 {acute over (Å)}) and using as eluents (A) 0.1% TFA in water and (B) 0.1% TFA in acetonitrile.

(146) Analytical HPLC was performed on a Acquity UPLC Waters Chromatograph with a BEH300 C.sub.4 Acquity Waters column 2.1×100 mm, 1.7 μm, at 45° C., using H.sub.2O, 0.1% TFA (A) and CH.sub.3CN, 0.1% TFA (B) as solvents. The peptides were characterized by electrospray mass spectrometry on an Acquity SQ Detector.

(147) Synthesis of Fmoc-4-(Dde-aminomethyl)-phenylalanine.

(148) ##STR00011##

(149) Fmoc-4-(Boc-aminomethyl)-phenylalanine was stirred in DCM/TFA 2/1 for 1 hour. The solvents were removed under reduced pressure and the residue was treated with diethyl ether to obtain a solid. The crude material obtained was dissolved in EtOH (19 mM), DIPEA (5 eq) and Dimedone (1.1 eq) were added to the reaction mixture. After 3 hours at 60° C. the solution was acidified with TFA to pH 4. The solvents were removed under reduced pressure and the residue was treated with AcOEt and washed with HCl 1N. The organic phase was washed with brine and dried over Na.sub.2SO.sub.4. The solvents were removed under reduced pressure and the final product was obtained as yellow oil which was further treated with Et.sub.2O to obtain a solid.

(150) The final compound was characterized on an Acquity UPLC Waters Chromatograph, with BEH300 C.sub.18 Acquity Waters 2.1×100 mm, 1.7 μm, at 45° C., using H.sub.2O, 0.1% TFA (A) and CH.sub.3CN, 0.1% TFA (B) as solvents and the following gradient: 10% A to 10% B in 1 min, 10% B over 90% B in 4 min, flow 0.4 mL/min. The protected amino acid was characterized by electrospray mass spectrometry on an Acquity SQ Detector (Mw found: 581.5 Da; Mw expected: 580.67 Da).

(151) Synthesis of Fmoc-Nle(1,2,3-triazole-5-PEG.sub.2-NH-ivDde

(152) ##STR00012##
was as follows.

(153) ##STR00013##

(154) TFA (0.013 mL, 0.175 mmol) was added to a stirred suspension of 2-(2-(prop-2-yn-1-yloxy)ethoxy)ethanamine (250 mg, 1.746 mmol) and 5,5-dimethyl-2-(3-methylbutanoyl)cyclohexane-1,3-dione (0.763 mL, 3.49 mmol) in ethanol (15 mL) at rt. The mixture was then refluxed for 24 hours and the solvent was rotary-evaporated. The residue was purified by silica gel chromatography (eluent: 0-15% methanol in DCM) to give 5,5-dimethyl-2-(3-methyl-1-((2-(2-(prop-2-yn-1-loxy)ethoxy)ethyl)amino)butylidene)cyclohexane-1,3-dione as light yellow oil in 82% yield.

(155) ##STR00014##

(156) A DCM (dichloromethane) solution of Fmoc-Lys(N3)—OH (500 mg, 1.268 mmol) was slowly added to a solution of NaHCO.sub.3 (1 eq) in water (3 mL/mmol NaHCO.sub.3). This was added to a solution of TBAB (tetrabutylammonium bromide; 1 eq) and allyl bromide (5.28 eq) in DCM (1.1 mL/mmol allyl bromide). The emulsion was stirred vigorously for 24 h at room temperature and was then extracted with DCM three times. The organic extracts were dried over MgSO.sub.4, and solvent was evaporated. The residue was purified by flash chromatography (SiO.sub.2, ethyl acetate:hexanes 0-50%) to obtain the product (S)-allyl 2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-6-azidohexanoate as white solids in 92% yield.

(157) ##STR00015##

(158) In glovebox, Cp*RuCl(PPh.sub.3).sub.2 (65.6 mg, 0.082 mmol) was added to a microwave tube with a septa cap. Dioxane (5 mL) was added. 5,5-dimethyl-2-(3-methyl-1-((2-(2-(prop-2-yn-1-loxy)ethoxy)ethyl)amino)butylidene)cyclohexane-1,3-dione (360 mg, 1.030 mmol) was dissolved in dioxane (2.5 mL) and added to the catalyst solution. Last, (S)-allyl 2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-6-azidohexanoate (448 mg, 1.030 mmol) was added in dioxane (2.5 mL) to the reaction mixture. The reaction was heated in an oil bath at 60° C. for 12 hours. The solvent was evaporated and the residue was purified by flash chromatography (80 g SiO.sub.2, ethyl acetate:hexanes 0-60% and then dichloromethane/methanol 0-20%) to give the product (S)-allyl 2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-6-(5-((2-(2-((1-(4,4-dimethyl-2,6-dioxocyclohexylidene)-3-methylbutyl)amino)ethoxy)ethoxy)methyl)-1H-1,2,3-triazol-1-yl)hexanoate as light yellow solid in 60% yield.

(159) Synthesis of Nle(1,2,3-triazole-4-C.sub.15)

(160) The structure of Nle(1,2,3-triazole-4-C.sub.15) wherein the linker is C.sub.15 alkyl is represented by

(161) ##STR00016##

(162) The GCG/GLP-1 receptor co-agonist peptides of the present invention of the following formula:

(163) (HsQGTFTSDK(N3)SKYLDARAAQDFVQWLLDT-NH.sub.2) (SEQ ID NO: 126) are connected to the linker through click chemistry described below

(164) ##STR00017##
HsQGTFTSD(Lys(N3)}SKYLDARAAQDFVQWLLD (SEQ ID NO: 127) (Biopeptek #173501-2, 100 mg, 0.030 mmol) and heptadec-1-yne (10.57 mg, 0.045 mmol) were dissolved in DMSO (6 mL). To this solution was added dropwise CuSO.sub.4*5H.sub.2O in water (3.0 mg/ml, 1.979 ml, 0.024 mmol) freshly mixed with aqueous sodium ascorbate (5904 μl, 0.119 mmol) in a water bath. The mixture was bubbled with N.sub.2 for 30 seconds, then sealed and shaken at room temperature for 18 hours. Then the reaction pH was adjusted to pH 3 using TFA. The resulting clear solution was filtered for purification (RPLC, column: Waters CSH C18 5μ, 19×150 mm; flowrate: 25 ml/min; gradient: water/ACN (acetonitrile) with 0.05% TFA 35-42%). Fractions containing the desired product were combined and lyophilized to give the title compound (10.1 mg, 2.70 μmol, 9.06% yield, purity 96%).

EXAMPLE 2

(165) Synthetic for peptide of sequence:

(166) TABLE-US-00015 (SEQ ID NO: 110) HUQGTFTSDYSKYLDURAAQDFVK(PEG2PEG2γEC18-OH)WL2DT-NH2
legend: U=aminoisobutyric acid; 2=L-methionine sulphone; PEG2=8-amino-3,6-dioxaoctanoic acid; γE=γ-glutamic acid; C18-OH (Octadecanedioic acid)=—CO—(CH2)16-COOH; tBu=tert butyl; and fmoc=9-fluorenylmethyl chloroformate.
The peptide was synthesized by solid phase synthesis using Fmoc/t-Bu chemistry on a peptide multisynthesizer Symphony (Protein Technologies Inc.) on a 180 μmol scale, using a Rink-amide PS resin (Novabiochem, loading 0.35 mmol/g). All the amino acids were dissolved at a 0.3 M concentration in DMF (dimethyl formamide). The amino acids were activated with equimolar amounts of HATU (O-(7-azabenzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium hexafluoro-phosphate) solution 0.3 M in DMF, and a 2-fold molar excess of DIEA (N,N-diisopropyl-ethylamine), solution 2M in NMP (N-methyl pyrrolidine). The acylation reactions were performed in general for 1 hour with a 5-fold excess of activated amino acid over the resin free amino groups with double 45 minutes acylation reactions performed from His.sup.1 to Thr.sup.7, from F.sup.22 to V.sup.23 and from D.sup.15 to Aib.sup.16. The side chain protecting groups were: tert-butyl for Glu, Ser, D-Ser (ser), Thr and Tyr; trityl for Asn, Gln and His; tert-butoxy-carbonyl for Lys, Trp; and, 2,2,4,6,7-pentamethyldihydro-benzofuran-5-sulfonyl for Arg. His was introduced as Boc-His(Trt)-OH at the end of the sequence assembly. Amino acid alpha-aminoisobutyric acid (Aib) was introduced by acylation of Fmoc-Aib-OH. Amino acid γ-Glu (γ-glutamic acid) was introduced by acylation of Fmoc-Glu-OtBu. Amino acid 2 (L-methionine-sulphone) was introduced by acylation of Fmoc-L-methionine-sulphone-COOH. The lysine used for linker-lipid derivatization was incorporated with a Dde [1-(4,4-dimethyl-2,6-dioxocyclohex-1-ylidene)ethyl] protecting group on the side chain of amino group. After the assembly the Dde protecting group of Lys (Dde) was removed by treatment of 2% hydrazine in DMF. The side chain of Lys was derivatized with Fmoc-PEG.sub.2 [8-(9-Fluorenylmethyloxycarbonyl-amino)-3,6-dioxaoctanoic acid], Fmoc-Glu-OtBu (γ-glutamic acid) using HOAt (1-Hydroxy-7-azabenzotriazole) and DIC as activators in DMF. The lipid diacid (Octadecanedioic acid) was introduced manually using HOAt and DIC as activators in NMP. At the end of the synthesis, the dry peptide-resin was individually treated with 35 mL of the cleavage mixture, 88% TFA (trifluoroacetic acid), 5% phenol, 2% triisopropylsilane and 5% water for 3 hours at room temperature. Each resin was filtered and then added to cold methyl-t-butyl ether in order to precipitate the peptide. After centrifugation, the peptide pellets were washed with fresh cold methyl-t-butyl ether to remove the organic scavengers. The process was repeated twice. Final pellets were dried, resuspended in H.sub.2O, 20% acetonitrile, and lyophilized. The crude peptide was purified by reverse-phase HPLC using preparative Waters Deltapak C4 (40×200 mm, 15 μm, 300 {acute over (Å)}) and using as eluents (A) 0.1% TFA in water and (B) 0.1% TFA in acetonitrile in order to obtain pure linear peptide. The final peptide was characterized on an Acquity UPLC Waters Chromatograph, with BEH130 C4 Acquity Waters 2.1×100 mm, 1.7 μm, at 45° C., using H2O, 0.1% TFA (A) and CH.sub.3CN, 0.1% TFA (B) as solvents. The peptide was characterized by electrospray mass spectrometry on a Acquity SQ Detector. (MW found: 4140.6 Da; MW expected: 4141.67).

EXAMPLE 3

(167) Activity of the peptides at the Glucagon receptor (GCGR) and GLP-1 receptor (GLP1R) was measured in a cAMP activity assay.

(168) GCG/GLP-1 receptor co-agonist peptides were dissolved in 100% DMSO (dimethyl sulfoxide) and serially diluted to generate 10 point titrations. The peptide solutions were then transferred into 384-well assay plates (150 nL/well). Assay ready frozen cells expressing human GLP1R or human GCGR were suspended in growth media consisting of DMEM medium (GIBCO®), 10% FBS (GIBCO®), 1×NEAA(GIBCO®), 1×P/S (GIBCO®), 10 ug/mL Blasticidin (GIBCO®) and 200 μg/mL Hygromycin (GIBCO®). Cells were then diluted in assay buffer consisting of PBS (GIBCO®), 7.5% BSA (Perkin Elmer), 100 μM RO 20-1724 (Sigma), with or without 20% human serum (MP Biomedical). The cell suspensions (15 μL) were then added to the assay plates containing the peptide solutions (30,000 cells/well for human GCGR; 10,000 cells/well for human GLP1R). The cells were incubated for 1 hour at room temperature in the dark. Production of cAMP was determined using HitHunter™ cAMPXS kits (DiscoverX) following manufacturer protocol. The plates were incubated for overnight at room temperature in the dark. Luminescence was measured using an EnVision Multilabel plate reader (Perkin Elmer). Native GLP-1 and Glucagon (Bachem) are used as control peptides. EC.sub.50 values were calculated using uses a 4 parameter logistic fit based on the Levenberg-Marquardt algorithm. The results are shown in Table 2.

(169) TABLE-US-00016 TABLE 2 GLP1R hGCGR/h SEQ ID Peptide GCGR EC50 EC50 human GLP1R NO: Name human (nM) (nM) (hTone) 1 TP565 5.420 2.260 2.4 2 TP579 >19.9 14.7 3 TP583 6.37 0.74 8.6 4 TP584 6 1.3 4.6 5 TP578 0.604 4.95 0.1 6 TP580 1.09 0.46 2.4 7 TP581 4.95 1.69 2.9 8 TP582 4.95 2.57 1.9 9 TP585 1.13 0.566 2.0 10 TP588 2.67 2.92 0.9 11 TP589 19.8 2.07 9.6 12 TP590 5.03 1.64 3.1 13 TP592 3.39 2.42 1.4 14 TP594 4.95 2.6 1.9 15 TP576 1.45 1.88 0.8 16 TP577 4.54 1.67 2.7 17 TP586 4.95 1.23 >4.0 18 TP587 2.49 0.41 >4.1 19 TP591 2.88 1.31 2.2 20 TP593 0.73 4.95 <0.15 21 TP595 0.4 >20 0.2 22 TP596 0.35 >20 0.2 23 TP599 3.47 2.21 1.6 24 TP600 1.75 0.57 3.1 25 TP601 1.8 8.5 0.2 26 TP602 2.17 0.36 6.3 27 TP603 1.88 1.17 1.6 28 TP605 3.75 1.98 1.9 29 TP606 1.01 1.84 0.5 30 TP607 0.57 1.06 0.5 31 TP610 0.19 >5 <0.01 32 TP611 0.2 >5 <0.02 33 TP612 2.04 4.95 <0.41 34 TP613 >5 3.34 >6 35 TP614 0.86 1.4 0.6 36 TP615 0.26 0.4 0.6 37 TP616 1.39 1.96 0.7 38 TP617 0.56 0.76 0.8 39 TP618 0.36 2.48 0.1 40 TP619 0.05 0.38 0.1 41 TP620 1.26 8.58 0.1 42 TP621 0.25 1.94 0.1 43 TP622 0.03 0.09 0.4 44 TP623 0.09 0.17 0.5 45 TP624 1 0.89 1.1 46 TP625 1.3 0.39 3.4 47 TP626 0.39 0.77 0.6 48 TP627 0.14 0.25 0.6 49 TP629 3.17 2.23 1.4 50 TP631 1.65 1.32 1.3 51 TP632 5.9 6.6 0.9 52 TP633 0.12 0.19 0.7 53 TP634 0.77 0.49 1.6 54 TP635 0.46 2.13 0.2 55 TP636 0.11 0.52 0.2 56 TP637 0.94 0.53 1.8 57 TP638 5.05 1.55 3.3 58 TP639 3.38 2.09 1.6 59 TP657 0.14 2.18 0.1 60 TP658 3.2 1.66 1.9 61 TP659 0.46 >4.9 NA 62 TP660 4.54 3.59 1.3 63 TP661 0.61 >19.8 NA 64 TP662 1.45 0.52 2.8 65 TP663 0.22 0.38 0.6 66 TP664 0.36 0.22 1.7 67 TP665 0.79 1.6 4.9 68 TP666 >19.8 >19.8 NA 69 TP667 >19.8 >19.8 NA 70 TP672 0.35 0.51 0.7 71 TP673 0.22 0.67 0.3 72 TP674 1.08 0.45 2.4 73 TP675 0.3 0.71 0.4 74 TP676 0.82 0.26 3.1 75 TP677 1.42 0.76 1.9 76 TP678 0.25 0.91 0.3 77 TP679 0.13 0.18 0.7 78 TP680 0.59 0.19 3.0 79 TP681 0.27 0.15 1.8 80 TP682 2.35 2.31 1.0 81 TP683 4.97 0.62 NA 82 TP685 4.97 1.12 7.9 83 TP693 1.1 >4.9 NA 84 TP699 >19.8 >19.8 NA 85 TP700 >4.9 0.35 NA 86 TP701 >4.9 0.18 NA 87 TP702 0.35 0.27 1.4 88 TP703 >19.8 >19.8 NA 89 TP704 0.27 0.37 0.7 90 TP705 1.48 0.41 3.6 91 TP712 0.25 0.45 0.6 92 TP713 1.52 0.71 2.2 93 TP735 0.895 0.648 1.4 94 TP736 3.19 1.145 2.8 95 TP737 1.56 2.67 0.6 96 TP811 0.21 0.26 0.8 97 TP812 0.2 0.088 2.3 98 TP813 0.21 0.14 1.6 99 TP814 0.156 0.078 2.0 100 TP815 0.084 0.12 0.7 101 TP825 0.07 0.036 2.0 102 TP826 0.67 0.253 2.7 103 TP827 0.133 0.081 1.6 104 TP828 0.228 0.587 0.4 109 TP564 1.250 1.220 1.0 110 TP575 0.595 0.866 0.7 111 TP597 0.74 0.61 1.2 112 TP598 1.14 0.5 2.3 113 TP604 1.09 0.47 2.3 114 TP443 0.18 0.1 1.8 115 TP608 0.21 0.23 0.9 116 TP609 0.36 0.42 0.9 117 TP628 0.86 1.39 0.6 118 TP630 0.89 0.29 3.0 119 TP640 0.69 0.84 0.8 120 TP829 1.06 0.68 1.56 121 TP830 0.14 0.22 0.64 122 TP831 0.14 0.08 1.66

EXAMPLE 4

(170) Diet induced obesity (DIO) mice have long been used as surrogates for humans in the study of the efficacy of anti-obesity compounds. The results obtained from such mice in the study of obesity compounds are translatable to humans (See for example, Nilsson et al. Acta Pharmacologia Sinica 33: 173-181 (2012), which is incorporated herein by reference in its entirety). Thus, DIO mice are useful surrogates for humans for the testing the efficacy of compounds intended to treat obesity.

(171) DIO mice are divided into groups of eight mice per group and the initial average body weight, food intake and basal glucose of each group were matched. Each group of mice is subcutaneously (sc) injected with a single dose of peptide or vehicle control. The administered doses may vary between 3 and 300 nmol/kg. Body weight and food intake are measured daily for four days after the initial dosing. Blood glucose is measured 5 hours post dose, and then daily for four days. A separate set of mice were treated with same dose of each peptide. Serial blood was taken at five hours, 24 hours, 48 hours and 72 hours post sc injection to measure drug exposure.

(172) While the present invention is described herein with reference to illustrated embodiments, it should be understood that the invention is not limited hereto. Those having ordinary skill in the art and access to the teachings herein will recognize additional modifications and embodiments within the scope thereof. Therefore, the present invention is limited only by the claims attached herein.