Compounds for the treatment of diabetes or obesity

Abstract

This invention relates to a novel urocortin-2 polypeptide active against corticotropin-releasing hormone receptor-2 (CRHR2); a pharmaceutical composition comprising the polypeptide; and a method of using the polypeptide to treat CRHR2-related conditions.

Description

Technical Field

[0001] This invention relates to polypeptides active against corticotropin-releasing hormone receptor-2 (CRHR2); pharmaceutical compositions comprising such polypeptides; and methods of using such polypeptides to treat related conditions. Prior Technology

[0002] Diabetes, obesity, and chronic kidney disease (CKD) are prevalent diseases worldwide, accompanied by significant health challenges. Although they are distinct diseases, they often occur together as comorbid conditions.

[0003] Diabetes is a metabolic disorder that occurs when the pancreas does not produce enough insulin or when the body needs more insulin than it produces, resulting in hyperglycemia or high blood sugar. In 2021, the U.S. Centers for Disease Control and Prevention (CDC) estimated that approximately 29.7 million people (8.9% of the U.S. population) were diagnosed with diabetes. More concerningly, an estimated 38.0% of U.S. adults aged 18 or older have prediabetes. Obesity is one of the major risk factors for diabetes.

[0004] Obesity is a medical condition that leads to an excessive accumulation of adipose tissue. Today, obesity is a global public health problem, with the CDC estimating it affects approximately 40% of the adult population. Furthermore, obesity is associated with high costs and serious, undesirable health outcomes and conditions. Some obese individuals may further exhibit a subset of obesity, such as sarcopenic obesity, a condition involving high body fat and low muscle mass. Both obesity and sarcopenic obesity can lead to metabolic problems such as type 2 diabetes mellitus (T2DM), hypertension, and cardiovascular disease. Therefore, quality weight loss is important for all obese individuals, meaning reducing adipose tissue with minimal or no impact on lean body mass.

[0005] In addition, diabetes and obesity are both risk factors for chronic kidney disease (CKD), a condition characterized by progressive loss of kidney function. Over time, individuals with CKD experience an increase in albuminuria, proteinuria, serum creatinine, and / or renal histopathological changes. For many patients, CKD eventually progresses to end-stage renal disease (ESRD), at which point dialysis or kidney transplantation is required.

[0006] Addressing the challenges associated with diabetes, obesity, and / or chronic kidney disease includes addressing one or more underlying factors that contribute to each patient’s specific condition (e.g., high blood sugar, overweight, excess fat mass relative to lean body mass).

[0007] Currently, patients with diabetes, obesity, and / or chronic kidney disease have numerous treatment options, including incretin agonists (such as liraglutide, tirzepatide, and semaglutide), amylase agonists (such as pramlintide), DPP-4 inhibitors, metformin, insulin, and many other options. However, currently available treatments may present several challenges in some patients, including short half-lives, the need for frequent dosing, undesirable side effects (such as gastrointestinal upset, vomiting, and / or hypoglycemia), immunogenicity, and / or the inability to effectively treat one or more synergistic factors associated with disease symptoms (such as hyperglycemia, overweight, or excess fat mass relative to lean body mass).

[0008] Human urocortin-2 (UCN2) is a 38-amino acid endogenous peptide (SEQ ID NO: 53). It is one of three known endogenous urocortins found in mammals (the other two being UCN1 and UCN3) and is part of the corticotropin-releasing hormone (CRH) family. Urocortin peptides are short-acting peptides that exert their effects through CRH receptors called CRHR1 and / or CRHR2. UCN2 has been reported to improve insulin sensitivity and glucose tolerance by acting as a ligand for the G protein-coupled receptor corticotropin-releasing hormone receptor 2 (CRHR2), and therefore, peptides active against the CRHR2 receptor have been investigated for the treatment of diabetes and related diseases. (Chen et al., 16580-16585 (2006)). However, although urocortin analogs are known in this technology (see, for example, US 10,894,817 (Alsina-Fernandez, Eli Lilly); WO 2023 / 285334 (Li, Novo Nordisk); WO 2023 / 161229 (Janiak, Corteria Pharmaceuticals)), no urocortin analogs have been authorized for medical use to date.

[0009] Therefore, there is a need for peptides active against the CRH2 receptor. Specifically, there is a need for therapeutic compounds that can treat obesity, diabetes, and / or CKD while preserving lean body mass. Furthermore, there is a need for compounds that include one or more of the following properties: good in vitro immunogenicity profile; selective efficacy against the CRH2 receptor; improved PK (half-life) compared to endogenous human urocortin; quality weight loss that preserves lean body mass compared to fat mass; and suitable exploitability properties (e.g., binding state). Summary of the Invention

[0010] This document provides polypeptides active against the CRH2 receptor for the treatment of diabetes, obesity, and / or CKD, and for addressing one or more of these challenges. According to one embodiment of the invention, this document provides a polypeptide of Formula I or a pharmaceutically acceptable salt thereof, comprising: X1X2X3X4X5X6IVTSX11DX13PX15X16X17LX19X20X21X22EQEX26X27EKX30X31QQAX35EX37X38EILAQV (SEQ ID NO: 68), where X1 is Pyr, E, γE, or does not exist. X2 is G or does not exist. X3 is G, S, or does not exist. X4 is S, P, G, or does not exist. X5 is S, P, or does not exist. X6 is G, S, P, or does not exist. X11 is L or αMeL, X13 is either V or D-Val. X15 is either T or I. X16 is either Aib or G. X17 is either L or αMeL. X19 is either Q or E. X20 is either K or I. X21 can be I, K, Aib, or L. X22 represents I or L. X26 is either R or K. X27 is either A or Q. X30 is either A or E. X31 is either R or K. X35 is either K or T. X37 is either A or N. X38 is either A or T. The N-terminal amino acid is N-acylated, N-acetylated, or N-methylated as appropriate, and the C-terminal amino acid is amylated as appropriate.

[0011] In some embodiments of Formula I, V at position 44 is the C-terminal amino acid. The C-terminus may contain amide, carboxylic acid, or other modifications. The N-terminal amino acid depends on the embodiment, but will always be the first amino acid in the sequence indicating the start of the polypeptide. Thus, in embodiments of Formula I, the N-terminal amino acid may be at positions X1, X2, X3, X4, X5, X6, or I at position 7. Furthermore, in some embodiments of Formula I, the N-terminus may contain modifications (e.g., acetylation, Ac; methylation, Me; acetylation; or other modifications). Additionally, if any amino acid at positions X1 to X6 is indicated as "absent," the next available amino acid will bind to the previously available amino acid. Thus, various embodiments of Formula I produce polypeptides containing different lengths, such as 38 to 44 amino acids.

[0012] In addition to the sequences described herein, the polypeptides described herein may also include one or more conserved amino acid substitutions, provided that the polypeptides are still active against CRHR2.

[0013] In some embodiments, the polypeptide or a pharmaceutically acceptable salt thereof further comprises a tool for increasing the half-life of the polypeptide. In some embodiments, the polypeptide or a pharmaceutically acceptable salt thereof further comprises a tool for increasing the half-life of the polypeptide via any amino acid having a functional group suitable for binding. In some embodiments, the amino acid having a functional group suitable for binding is γE, E, or K. In some embodiments, the functional group suitable for binding may bind to phosphonates, tetrazolium, sulfonates, branched fatty acids, peptides, proteins (such as VHH, Fc, or monoclonal antibodies) or protein fragments (such as Fab, scFv, Fv, or scFab (single-chain Fab)). In some embodiments, the fatty acid, linker, protein, or protein fragment may further act as an albumin binder and provide the potential to further generate a long-acting compound.

[0014] In some embodiments, the tool is a fatty acid.

[0015] In some embodiments, the amino acid having the functional group for binding is K, and the binding is with the ε-amino group of K. In some embodiments, the polypeptide further comprises a fatty acid bound to K at X35, wherein the fatty acid is bound to the ε-amino group of K via a direct bond or linker between K and the fatty acid. In some embodiments, the amino acid having the functional group for binding is γE at X1, and the binding is via an amide bond between the α-amino group of γE and the carboxyl group of the fatty acid. In some embodiments, the tool for increasing the half-life of the polypeptide is a fatty acid, wherein the fatty acid is bound to the polypeptide at the amino acid having the functional group for binding via a direct bond or linker between the amino acid and the fatty acid.

[0016] In some embodiments, the N-terminal amino acid is N-acylated via a fatty acid, which is bound to the N-terminal amino acid via a direct bond or linker between the amino acid and the fatty acid. In some such embodiments, the linker-fatty acid is bound to the peptide after peptide synthesis. In other such embodiments, the linker-fatty acid is bound to the amino acid (e.g., in some embodiments, with E or γE) before being added to the peptide.

[0017] In some embodiments, the polypeptide is bound to the fatty acid via a linker between the amino acid and the fatty acid. Therefore, in some embodiments, the polypeptide is bound to the fatty acid via a direct bond between the amino acid and the fatty acid. In other embodiments, the polypeptide is bound to the fatty acid via a linker between the amino acid and the fatty acid. In some embodiments, the fatty acid is a C16-C22 fatty acid. In some embodiments, the binding of the linker to the C16-C22 fatty acid is referred to as linker-fatty acid.

[0018] Examples of saturated C16-C22 fatty acids used in this document include (but are not limited to) palmitic acid (hexadecanoic acid) (C16 monocarboxylic acid), hexadecanoic acid (C16 diacid), pearlitic acid (heptadecanoic acid) (C17 monocarboxylic acid), heptadecanoic acid (C17 diacid), stearic acid (C18 monocarboxylic acid), octadecanoic acid (C18 diacid), nonadecanoic acid (nonadecanoic acid) (C19 monocarboxylic acid), nonadecanoic acid (C19 diacid), arachidic acid (eicosanoic acid) (C20 monocarboxylic acid), eicosanoic acid (C20 diacid), icosanoic acid (icosanoic acid) (C21 monocarboxylic acid), icosanoic acid (icosanoic acid) (C22 monocarboxylic acid), and icosanoic acid (C22 diacid), including their branched chains and substituted derivatives.

[0019] In some cases, polypeptides contain linkers. Linkers may have one or more (2-[2-(2-amino-ethoxy)-ethoxy]-acetyl moieties or εK, in combination with one or more amino acids, as appropriate. Where a linker includes at least one amino acid, the amino acid may be an E or γE amino acid residue. In some cases, a linker may include one, two, three, four, or five E or γE amino acid residues, including their D-type. For example, a linker may include one, two, three, or four γE amino acid residues. Alternatively, a linker may include one to five amino acid residues (e.g., E or γE amino acids) in combination with one to five (2-[2-(2-amino-ethoxy)-ethoxy]-acetyl) (“AEEA”) or one to five εK moieties. Specifically, a linker may be one to five E or γE amino acids in combination with one to five (2-[2-(2-amino-ethoxy)-ethoxy]-acetyl) (“AEEA”) moieties. ("AEEA") or a combination of one to five εK moieties, or a combination of one to five E or γE amino acids with one to five εK moieties. In some cases, the linker may be a combination of one or two or three γE amino acids with one or two (2-[2-(2-amino-ethoxy)-ethoxy]-acetyl) or εK moieties.

[0020] In certain embodiments, the polypeptide or a pharmaceutically acceptable salt thereof further comprises a linker-fatty acid, wherein the linker-fatty acid is selected from the group consisting of: (2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-PKE-γE-CO-(CH2)18-CO2H, Ahx-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-γE-CO-(CH2)18-CO2H, γE-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-Ahx-CO-(CH2)18-CO2H, γE-GKEKEKE-CO-(CH2)18-CO2H, G-(βA)3- γE-CO-(CH2)18-CO2H, G-(βA)4-γE-CO-(CH2)18-CO2H, (2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-KEKEKE-γE-CO-(CH2)18-CO2H, βA-S-βA-S-γE-CO-(CH2)18-CO2H, γE-PKE-Ahx-CO-(CH2)18-CO2H, γE-PKE-εK-CO-(CH2)18-CO2H, γE-PPP-εK-CO-(CH2)18-CO2H, γE-PPPPPP-CO-(CH2)18-CO2H, γE-PEP EPE-CO-(CH2)18-CO2H, γE-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)2-εK-CO-(CH2)18-CO2H, γE-εK-γE-εK-CO-(CH2)18-CO2H, γE-εK-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-εK-CO-(CH2)18-CO2H, γE-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-εK-CO-(CH2)16-CO2H, 2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-εK-CO-(CH2)18- CO2H, (2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-K-CO-(CH2)18-CO2H, (2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-εK-CO-(CH2)16-CO2H, (2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-Dap-CO-(CH2)18-CO2H, (2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)2-γE-CO-(CH2)18-CO2H, γE-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-γE-CO-(CH2)18-CO2H,γE2-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-CO-(CH2)18-CO2H, 2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-εK-γE-CO-(CH2)18-CO2, 2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-γE-εK-CO-(CH2)18-CO2H, εK-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-γE-CO-(CH2)18-CO2H, εK-γE-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-γE-CO-(CH2)18-CO2H, εK-γE-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)- γE-εK-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-CO-(CH2)18-CO2H, γE-γE-εK-CO-(CH2)18-CO2H, γE-εK-γE-CO-(CH2)18-CO2H, εK-γE-γE-CO-(CH2)18-CO2H, K(GKEKEKE-CO-(CH2)18-CO2H), 2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-γE2-CO-(CH2)18-CO2H, γE- (2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-εK-CO-(CH2)18-CO2H, 2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)2γE-C20-OH, and γE-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-εK-CO-(CH2)18-CO2H.

[0021] In some embodiments, the N-terminal amino acid is N-acetylated. In some embodiments, the N-terminal amino acid is N-methylated. In some embodiments, the C-terminal amino acid is amylated.

[0022] In one embodiment of Formula I, X1 is absent, X2 is absent, X3 is G, X4 is S, X5 is P, X6 is S, X11 is αMeL, X13 is V, X15 is T, X16 is Aib, X17 is αMeL, X19 is Q, X20 is K, X21 is I or L, X22 is L, X26 is R, X27 is A, X30 is A, X31 is R, X35 is K, X37 is A, and X38 is A. In this type of embodiment, the polypeptide comprises a sequence of 42 amino acids.

[0023] Therefore, in some embodiments, this document provides a polypeptide of formula Ia or a pharmaceutically acceptable salt thereof, comprising: GSPSIVTS αMeL DVPT Aib αMeL LQKX21LEQERAEKARQQAKEAAEILAQV (SEQ ID NO: 69), where X21 is either I or L [where X21 is located at position 19 of the polypeptide when X1 and X2 are absent]. The N-terminal amino acid is N-acetylated as appropriate, and the C-terminal amino acid is amylated as appropriate.

[0024] In some embodiments of Formula Ia, X21 is I. In some embodiments, the polypeptide comprises SEQ ID NO: 20. In another embodiment, the polypeptide is SEQ ID NO: 20. Alternatively, the polypeptide may have at least about 90% to 97% sequence similarity to SEQ ID NO: 20. Alternatively, the polypeptide may have at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, or 97% sequence similarity to SEQ ID NO: 20.

[0025] In a specific embodiment of formula Ia comprising a polypeptide or a pharmaceutically acceptable salt thereof, X21 is I and the polypeptide further comprises a linker-fatty acid. In a specific embodiment, the linker-fatty acid is γE-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-εK-CO-(CH2)18-CO2H. In a specific embodiment of formula Ia comprising a polypeptide or a pharmaceutically acceptable salt thereof, K at X35 is bound to the linker-fatty acid. In another embodiment, the polypeptide comprises SEQ ID NO: 1. In another embodiment, the polypeptide is SEQ ID NO: 1. Alternatively, the polypeptide may have at least about 90% to 97% sequence similarity to SEQ ID NO: 1. Alternatively, the polypeptide may have at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, or 97% sequence similarity to SEQ ID NO: 1.

[0026] In a specific embodiment of Formula Ia comprising a polypeptide or a pharmaceutically acceptable salt thereof, X21 is L and the polypeptide further comprises a linker-fatty acid. In a specific embodiment, the linker-fatty acid is γE-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-εK-CO-(CH2)18-CO2H. In a specific embodiment of Formula Ia comprising a polypeptide or a pharmaceutically acceptable salt thereof, K at X35 is bound to the linker-fatty acid. In another embodiment, the polypeptide comprises SEQ ID NO: 2. In another embodiment, the polypeptide is SEQ ID NO: 2. Alternatively, the polypeptide may have at least about 90% to 97% sequence similarity to SEQ ID NO: 2. Alternatively, the polypeptide may have at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, or 97% sequence similarity to SEQ ID NO: 2.

[0027] In a specific embodiment of formula Ia comprising a polypeptide or a pharmaceutically acceptable salt thereof, X21 is I and the polypeptide further comprises a linker-fatty acid. In a specific embodiment, the linker-fatty acid is 2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-PKE-γE-CO-(CH2)18-CO2H. In a specific embodiment of formula Ia comprising a polypeptide or a pharmaceutically acceptable salt thereof, K at X35 is bound to the linker-fatty acid. In another embodiment, the polypeptide comprises SEQ ID NO: 3. In another embodiment, the polypeptide is SEQ ID NO: 3. Alternatively, the polypeptide may have at least about 90% to 97% sequence similarity to SEQ ID NO: 3. Alternatively, the polypeptide may have at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, or 97% sequence similarity to SEQ ID NO: 3.

[0028] In a specific embodiment comprising a polypeptide or a pharmaceutically acceptable salt thereof of formula Ia, X21 is I and the polypeptide further comprises a linker-fatty acid. In a specific embodiment, the linker-fatty acid is Ahx-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-γE-CO-(CH2)18-CO2H. In a specific embodiment comprising a polypeptide or a pharmaceutically acceptable salt thereof of formula Ia, K at X35 is bound to the linker-fatty acid. In another embodiment, the polypeptide comprises SEQ ID NO: 4. In another embodiment, the polypeptide is SEQ ID NO: 4. Alternatively, the polypeptide may have at least about 90% to 97% sequence similarity to SEQ ID NO: 4. Alternatively, the polypeptide may have at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, or 97% sequence similarity to SEQ ID NO: 4.

[0029] In a specific embodiment of formula Ia comprising a polypeptide or a pharmaceutically acceptable salt thereof, X21 is I and the polypeptide further comprises a linker-fatty acid. In a specific embodiment, the linker-fatty acid is γE-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-Ahx-CO-(CH2)18-CO2H. In a specific embodiment of formula Ia comprising a polypeptide or a pharmaceutically acceptable salt thereof, K at X35 is bound to the linker-fatty acid. In another embodiment, the polypeptide comprises SEQ ID NO: 5. In another embodiment, the polypeptide is SEQ ID NO: 5. Alternatively, the polypeptide may have at least about 90% to 97% sequence similarity to SEQ ID NO: 5. Alternatively, the polypeptide may have at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, or 97% sequence similarity to SEQ ID NO: 5.

[0030] In a specific embodiment comprising a polypeptide or a pharmaceutically acceptable salt thereof of formula Ia, X21 is I and the polypeptide further comprises a linker-fatty acid. In a specific embodiment, the linker-fatty acid is γE-GKEKEKE-CO-(CH2)18-CO2H. In a specific embodiment comprising a polypeptide or a pharmaceutically acceptable salt thereof of formula Ia, K at X35 is bound to the linker-fatty acid. In another embodiment, the polypeptide comprises SEQ ID NO: 6. In another embodiment, the polypeptide is SEQ ID NO: 6. Alternatively, the polypeptide may have at least about 90% to 97% sequence similarity to SEQ ID NO: 6. Alternatively, the polypeptide may have at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, or 97% sequence similarity to SEQ ID NO: 6.

[0031] In a specific embodiment comprising a polypeptide or a pharmaceutically acceptable salt thereof of formula Ia, X21 is I and the polypeptide further comprises a linker-fatty acid. In a specific embodiment, the linker-fatty acid is G-(βA)3-γE-CO-(CH2)18-CO2H. In a specific embodiment comprising a polypeptide or a pharmaceutically acceptable salt thereof of formula Ia, K at X35 is bound to the linker-fatty acid. In another embodiment, the polypeptide comprises SEQ ID NO: 7. In another embodiment, the polypeptide is SEQ ID NO: 7. Alternatively, the polypeptide may have at least about 90% to 97% sequence similarity to SEQ ID NO: 7. Alternatively, the polypeptide may have at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, or 97% sequence similarity to SEQ ID NO: 7.

[0032] In a specific embodiment comprising a polypeptide or a pharmaceutically acceptable salt thereof of formula Ia, X21 is I and the polypeptide further comprises a linker-fatty acid. In a specific embodiment, the linker-fatty acid is G-(βA)4-γE-CO-(CH2)18-CO2H. In a specific embodiment comprising a polypeptide or a pharmaceutically acceptable salt thereof of formula Ia, K at X35 is bound to the linker-fatty acid. In another embodiment, the polypeptide comprises SEQ ID NO: 8. In another embodiment, the polypeptide is SEQ ID NO: 8. Alternatively, the polypeptide may have at least about 90% to 97% sequence similarity to SEQ ID NO: 8. Alternatively, the polypeptide may have at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, or 97% sequence similarity to SEQ ID NO: 8.

[0033] In a specific embodiment of formula Ia comprising a polypeptide or a pharmaceutically acceptable salt thereof, X21 is I and the polypeptide further comprises a linker-fatty acid. In a specific embodiment, the linker-fatty acid is 2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-KEKEKE-γE-CO-(CH2)18-CO2H. In a specific embodiment of formula Ia comprising a polypeptide or a pharmaceutically acceptable salt thereof, K at X35 is bound to the linker-fatty acid. In another embodiment, the polypeptide comprises SEQ ID NO: 9. In another embodiment, the polypeptide is SEQ ID NO: 9. Alternatively, the polypeptide may have at least about 90% to 97% sequence similarity to SEQ ID NO: 9. Alternatively, the polypeptide may have at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, or 97% sequence similarity to SEQ ID NO: 9.

[0034] In a specific embodiment comprising a polypeptide or a pharmaceutically acceptable salt thereof of formula Ia, X21 is I and the polypeptide further comprises a linker-fatty acid. In a specific embodiment, the linker-fatty acid is βA-S-βA-S-γE-CO-(CH2)18-CO2H. In a specific embodiment comprising a polypeptide or a pharmaceutically acceptable salt thereof of formula Ia, K at X35 is bound to the linker-fatty acid. In another embodiment, the polypeptide comprises SEQ ID NO: 10. In another embodiment, the polypeptide is SEQ ID NO: 10. Alternatively, the polypeptide may have at least about 90% to 97% sequence similarity to SEQ ID NO: 10. Alternatively, the polypeptide may have at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, or 97% sequence similarity to SEQ ID NO: 10.

[0035] In a specific embodiment comprising a polypeptide or a pharmaceutically acceptable salt thereof of formula Ia, X21 is I and the polypeptide further comprises a linker-fatty acid. In a specific embodiment, the linker-fatty acid is γE-PKE-Ahx-CO-(CH2)18-CO2H. In a specific embodiment comprising a polypeptide or a pharmaceutically acceptable salt thereof of formula Ia, K at X35 is bound to the linker-fatty acid. In another embodiment, the polypeptide comprises SEQ ID NO: 11. In another embodiment, the polypeptide is SEQ ID NO: 11. Alternatively, the polypeptide may have at least about 90% to 97% sequence similarity to SEQ ID NO: 11. Alternatively, the polypeptide may have at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, or 97% sequence similarity to SEQ ID NO: 11.

[0036] In a specific embodiment of Formula Ia comprising a polypeptide or a pharmaceutically acceptable salt thereof, X21 is L and the polypeptide further comprises a linker-fatty acid. In a specific embodiment, the linker-fatty acid is γE-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-εK-CO-(CH2)16-CO2H. In a specific embodiment of Formula Ia comprising a polypeptide or a pharmaceutically acceptable salt thereof, K at X35 is bound to the linker-fatty acid. In another embodiment, the polypeptide comprises SEQ ID NO: 12. In another embodiment, the polypeptide is SEQ ID NO: 12. Alternatively, the polypeptide may have at least about 90% to 97% sequence similarity to SEQ ID NO: 12. Alternatively, the polypeptide may have at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, or 97% sequence similarity to SEQ ID NO: 12.

[0037] In a specific embodiment comprising a polypeptide or a pharmaceutically acceptable salt thereof of formula Ia, X21 is I and the polypeptide further comprises a linker-fatty acid. In a specific embodiment, the linker-fatty acid is γE-PKE-εK-CO-(CH2)18-CO2H. In a specific embodiment comprising a polypeptide or a pharmaceutically acceptable salt thereof of formula Ia, K at X35 is bound to the linker-fatty acid. Alternatively, the polypeptide may have at least about 90% to 97% sequence similarity to SEQ ID NO: 13. Alternatively, the polypeptide may have at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, or 97% sequence similarity to SEQ ID NO: 13.

[0038] In a specific embodiment comprising a polypeptide or a pharmaceutically acceptable salt thereof of formula Ia, X21 is I and the polypeptide further comprises a linker-fatty acid. In a specific embodiment, the linker-fatty acid is γE-PPP-εK-CO-(CH2)18-CO2H. In a specific embodiment comprising a polypeptide or a pharmaceutically acceptable salt thereof of formula Ia, K at X35 is bound to the linker-fatty acid. In another embodiment, the polypeptide comprises SEQ ID NO: 14. In another embodiment, the polypeptide is SEQ ID NO: 14. Alternatively, the polypeptide may have at least about 90% to 97% sequence similarity to SEQ ID NO: 14. Alternatively, the polypeptide may have at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, or 97% sequence similarity to SEQ ID NO: 14.

[0039] In a specific embodiment comprising a polypeptide or a pharmaceutically acceptable salt thereof of formula Ia, X21 is I and the polypeptide further comprises a linker-fatty acid. In a specific embodiment, the linker-fatty acid is γE-PPPPPP-CO-(CH2)18-CO2H. In a specific embodiment comprising a polypeptide or a pharmaceutically acceptable salt thereof of formula Ia, K at X35 is bound to the linker-fatty acid. In another embodiment, the polypeptide comprises SEQ ID NO: 15. In another embodiment, the polypeptide is SEQ ID NO: 15. Alternatively, the polypeptide may have at least about 90% to 97% sequence similarity to SEQ ID NO: 15. Alternatively, the polypeptide may have at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, or 97% sequence similarity to SEQ ID NO: 15.

[0040] In a specific embodiment comprising a polypeptide or a pharmaceutically acceptable salt thereof of formula Ia, X21 is I and the polypeptide further comprises a linker-fatty acid. In a specific embodiment, the linker-fatty acid is γE-PEPEPE-CO-(CH2)18-CO2H. In a specific embodiment comprising a polypeptide or a pharmaceutically acceptable salt thereof of formula Ia, K at X35 is bound to the linker-fatty acid. In another embodiment, the polypeptide comprises SEQ ID NO: 16. In another embodiment, the polypeptide is SEQ ID NO: 16. Alternatively, the polypeptide may have at least about 90% to 97% sequence similarity to SEQ ID NO: 16. Alternatively, the polypeptide may have at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, or 97% sequence similarity to SEQ ID NO: 16.

[0041] In a specific embodiment of formula Ia comprising a polypeptide or a pharmaceutically acceptable salt thereof, X21 is I and the polypeptide further comprises a linker-fatty acid. In a specific embodiment, the linker-fatty acid is γE-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)2-εK-CO-(CH2)18-CO2H. In a specific embodiment of formula Ia comprising a polypeptide or a pharmaceutically acceptable salt thereof, K at X35 is bound to the linker-fatty acid. In another embodiment, the polypeptide comprises SEQ ID NO: 17. In another embodiment, the polypeptide is SEQ ID NO: 17. Alternatively, the polypeptide may have at least about 90% to 97% sequence similarity to SEQ ID NO: 17. Alternatively, the polypeptide may have at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, or 97% sequence similarity to SEQ ID NO: 17.

[0042] In a specific embodiment of formula Ia comprising a polypeptide or a pharmaceutically acceptable salt thereof, X21 is I and the polypeptide further comprises a linker-fatty acid. In a specific embodiment, the linker-fatty acid is γE-εK-γE-εK-CO-(CH2)18-CO2H. In a specific embodiment of formula Ia comprising a polypeptide or a pharmaceutically acceptable salt thereof, K at X35 is bound to the linker-fatty acid. Alternatively, the polypeptide may have at least about 90% to 97% sequence similarity to SEQ ID NO: 18. Alternatively, the polypeptide may have at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, or 97% sequence similarity to SEQ ID NO: 18.

[0043] In a specific embodiment of formula Ia comprising a polypeptide or a pharmaceutically acceptable salt thereof, X21 is I and the polypeptide further comprises a linker-fatty acid. In a specific embodiment, the linker-fatty acid is γE-εK-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-εK-CO-(CH2)18-CO2H. In a specific embodiment of formula Ia comprising a polypeptide or a pharmaceutically acceptable salt thereof, K at X35 is bound to the linker-fatty acid. In another embodiment, the polypeptide comprises SEQ ID NO: 19. In another embodiment, the polypeptide is SEQ ID NO: 19. Alternatively, the polypeptide may have at least about 90% to 97% sequence similarity to SEQ ID NO: 19. Alternatively, the polypeptide may have at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, or 97% sequence similarity to SEQ ID NO: 19.

[0044] In a specific embodiment of formula Ia comprising a polypeptide or a pharmaceutically acceptable salt thereof, X21 is I and the polypeptide further comprises a linker-fatty acid. In a specific embodiment, the linker-fatty acid is γE-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-εK-CO-(CH2)16-CO2H. In a specific embodiment of formula Ia comprising a polypeptide or a pharmaceutically acceptable salt thereof, K at X35 is bound to the linker-fatty acid. In another embodiment, the polypeptide comprises SEQ ID NO: 21. In another embodiment, the polypeptide is SEQ ID NO: 21. Alternatively, the polypeptide may have at least about 90% to 97% sequence similarity to SEQ ID NO: 21. Alternatively, the polypeptide may have at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, or 97% sequence similarity to SEQ ID NO: 21.

[0045] In another embodiment of Formula I, X1 is Pyr, X2 is G, X3 is S, X4 is P, X5 is S, X6 is G, X11 is αMeL, X13 is V, X15 is T, X16 is Aib, X17 is αMeL, X19 is Q, X20 is K, X21 is I, X22 is L, X26 is R, X27 is A, X30 is A, X31 is R, X35 is K, X37 is A, and X38 is A. In such embodiments, the polypeptide comprises a sequence of 44 amino acids.

[0046] Therefore, in some embodiments, this document provides a polypeptide of formula Ib or a pharmaceutically acceptable salt thereof, comprising:

[0047] Pyr GSPSGIVTS αMeL DVPT Aib αMeL LQKILEQERAEKARQQAKEAAEILAQV (SEQ ID NO: 70),

[0048] The C-terminal amino acid is acetaminated as appropriate.

[0049] In another embodiment, the polypeptide comprises SEQ ID NO: 23. In another embodiment, the polypeptide is SEQ ID NO: 23. Alternatively, the polypeptide may have at least about 90% to 97% sequence similarity to SEQ ID NO: 23. Alternatively, the polypeptide may have at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, or 97% sequence similarity to SEQ ID NO: 23.

[0050] In a specific embodiment comprising a polypeptide or a pharmaceutically acceptable salt thereof of formula Ib, the polypeptide further comprises a linker-fatty acid. In a specific embodiment, the linker-fatty acid is γE-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-εK-CO-(CH2)18-CO2H. In a specific embodiment comprising a polypeptide or a pharmaceutically acceptable salt thereof of formula Ib, K at X35 is bound to the linker-fatty acid. In another embodiment, the polypeptide comprises SEQ ID NO: 22. In another embodiment, the polypeptide is SEQ ID NO: 22. Alternatively, the polypeptide may have at least about 90% to 97% sequence similarity to SEQ ID NO: 22. Alternatively, the polypeptide may have at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, or 97% sequence similarity to SEQ ID NO: 22.

[0051] In a specific embodiment comprising a polypeptide or a pharmaceutically acceptable salt thereof of formula Ib, the polypeptide further comprises a linker-fatty acid. In a specific embodiment, the linker-fatty acid is γE-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-εK-CO-(CH2)16-CO2H. In a specific embodiment comprising a polypeptide or a pharmaceutically acceptable salt thereof of formula Ib, K at X35 is bound to the linker-fatty acid. In another embodiment, the polypeptide comprises SEQ ID NO: 24. In another embodiment, the polypeptide is SEQ ID NO: 24. Alternatively, the polypeptide may have at least about 90% to 97% sequence similarity to SEQ ID NO: 24. Alternatively, the polypeptide may have at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, or 97% sequence similarity to SEQ ID NO: 24.

[0052] In another embodiment of Equation I, X1 is γE, E, or does not exist; X2 does not exist; X3 is G; X4 is S; X5 is P; X6 is S; X11 is αMeL; X13 is V; X15 is T; X16 is Aib; X17 is αMeL; X19 is Q or E; X20 is K; X21 is I; X22 is L; X26 is R; X27 is A; X30 is A; X31 is R; X35 is T; X37 is A; and X38 is A.

[0053] In another embodiment of Formula I, X1 is absent, X2 is absent, X3 is G, X4 is S, X5 is P, X6 is S, X11 is αMeL, X13 is V, X15 is T, X16 is Aib, X17 is αMeL, X19 is Q, X20 is K, X21 is I, X22 is L, X26 is R, X27 is A, X30 is A, X31 is R, X35 is T, X37 is A, and X38 is A. In such embodiments, the polypeptide comprises a sequence of 42 amino acids.

[0054] In another embodiment, the polypeptide comprises SEQ ID NO: 27. In another embodiment, the polypeptide is SEQ ID NO: 27. Alternatively, the polypeptide may have at least about 90% to 97% sequence similarity to SEQ ID NO: 27. Alternatively, the polypeptide may have at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, or 97% sequence similarity to SEQ ID NO: 27.

[0055] In another embodiment of Formula I, X1 is γE or E, X2 is absent, X3 is G, X4 is S, X5 is P, X6 is S, X11 is αMeL, X13 is V, X15 is T, X16 is Aib, X17 is αMeL, X19 is Q or E, X20 is K, X21 is I, X22 is L, X26 is R, X27 is A, X30 is A, X31 is R, X35 is T, X37 is A, and X38 is A. In such embodiments, the polypeptide comprises a sequence of 43 amino acids.

[0056] Therefore, in some embodiments, this document provides a polypeptide of formula Ic or a pharmaceutically acceptable salt thereof, comprising: X1GSPSIVTS αMeL DVPT Aib αMeLLX19KILEQERAEKARQQATEAAEILAQV (SEQ ID NO: 71), where X1 is γE or E, X19 is either Q or E [where X19 is at position 18 of the polypeptide when X2 is absent], The C-terminal amino acid is acetaminated as appropriate.

[0057] In a specific embodiment comprising a polypeptide or a pharmaceutically acceptable salt thereof of formula Ic, X1 is γE, X19 is Q, and the linker-fatty acid is (2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-εK-CO-(CH2)18-CO2H. In another embodiment, the polypeptide comprises SEQ ID NO: 25. In another embodiment, the polypeptide is SEQ ID NO: 25. Alternatively, the polypeptide may have at least about 90% to 97% sequence similarity to SEQ ID NO: 25. Alternatively, the polypeptide may have at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, or 97% sequence similarity to SEQ ID NO: 25.

[0058] In a specific embodiment comprising a polypeptide or a pharmaceutically acceptable salt thereof of formula Ic, X1 is γE, X19 is Q, and the linker-fatty acid is (2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-K-CO-(CH2)18-CO2H. In another embodiment, the polypeptide comprises SEQ ID NO: 26. In another embodiment, the polypeptide is SEQ ID NO: 26. Alternatively, the polypeptide may have at least about 90% to 97% sequence similarity to SEQ ID NO: 26. Alternatively, the polypeptide may have at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, or 97% sequence similarity to SEQ ID NO: 26.

[0059] In a specific embodiment comprising a polypeptide or a pharmaceutically acceptable salt thereof of formula Ic, X1 is γE, X19 is Q, and the linker-fatty acid is (2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-εK-CO-(CH2)16-CO2H. In another embodiment, the polypeptide comprises SEQ ID NO: 28. In another embodiment, the polypeptide is SEQ ID NO: 28. Alternatively, the polypeptide may have at least about 90% to 97% sequence similarity to SEQ ID NO: 28. Alternatively, the polypeptide may have at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, or 97% sequence similarity to SEQ ID NO: 28.

[0060] In a specific embodiment comprising a polypeptide or a pharmaceutically acceptable salt thereof of formula Ic, X1 is γE, X19 is E, and the linker-fatty acid is (2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-Dap-CO-(CH2)18-CO2H. In another embodiment, the polypeptide comprises SEQ ID NO: 52. In another embodiment, the polypeptide is SEQ ID NO: 52. Alternatively, the polypeptide may have at least about 90% to 97% sequence similarity to SEQ ID NO: 52. Alternatively, the polypeptide may have at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, or 97% sequence similarity to SEQ ID NO: 52.

[0061] In another embodiment of Formula I, X1 is absent, X2 is absent, X3 is absent, X4 is absent, X5 is absent, X6 is absent, X11 is αMeL, X13 is D-Val, X15 is T or I, X16 is Aib or G, X17 is αMeL, X19 is Q, X20 is I, X21 is K, X22 is L, X26 is R, X27 is A, X30 is A, X31 is R, X35 is K, X37 is N, and X38 is T. In such embodiments, the polypeptide comprises a sequence of 38 amino acids.

[0062] Therefore, in some embodiments, this document provides a polypeptide of formula Id or a pharmaceutically acceptable salt thereof, comprising... IVTS αMeL D D-Val PX15X16αMeLLQIKLEQERAEKARQQAKENTEILAQV (SEQ ID NO: 72), where X15 is either T or I [where X1 to X6 are absent, X15 is located at position 9 of the polypeptide], X16 is either G or Aib [where X1 to X6 are absent, X16 is at position 10 of the polypeptide], The N-terminal amino acid is N-acetylated as appropriate, and the C-terminal amino acid is amylated as appropriate.

[0063] In a specific embodiment comprising a polypeptide or a pharmaceutically acceptable salt thereof of formula Id, X15 is T, X16 is G, and the polypeptide further comprises a linker-fatty acid. In a specific embodiment, the linker-fatty acid is 2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)2-γE-CO-(CH2)18-CO2H. In a specific embodiment comprising a polypeptide or a pharmaceutically acceptable salt thereof of formula Id, K at X35 is bound to the linker-fatty acid. In another embodiment, the polypeptide is SEQ ID NO: 29. Alternatively, the polypeptide may have at least about 90% to 97% sequence similarity to SEQ ID NO: 29. Alternatively, the polypeptide may have at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, or 97% sequence similarity to SEQ ID NO: 29.

[0064] In a specific embodiment comprising a polypeptide or a pharmaceutically acceptable salt thereof of formula Id, X15 is T, X16 is G, and the polypeptide further comprises a linker-fatty acid. In a specific embodiment, the linker-fatty acid is γE-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-γE-CO-(CH2)18-CO2H. In a specific embodiment comprising a polypeptide or a pharmaceutically acceptable salt thereof of formula Id, K at X35 is bound to the linker-fatty acid. In another embodiment, the polypeptide comprises SEQ ID NO: 30. In another embodiment, the polypeptide is SEQ ID NO: 30. Alternatively, the polypeptide may have at least about 90% to 97% sequence similarity to SEQ ID NO: 30. Alternatively, the polypeptide may have at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, or 97% sequence similarity to SEQ ID NO: 30.

[0065] In a specific embodiment comprising a polypeptide or a pharmaceutically acceptable salt thereof of formula Id, X15 is T, X16 is G, and the polypeptide further comprises a linker-fatty acid. In a specific embodiment, the linker-fatty acid is γE2-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-CO-(CH2)18-CO2H. In a specific embodiment comprising a polypeptide or a pharmaceutically acceptable salt thereof of formula Id, K at X35 is bound to the linker-fatty acid. In another embodiment, the polypeptide comprises SEQ ID NO: 31. In another embodiment, the polypeptide is SEQ ID NO: 31. Alternatively, the polypeptide may have at least about 90% to 99% sequence similarity to SEQ ID NO: 31. Alternatively, the polypeptide may have at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence similarity to SEQ ID NO: 31.

[0066] In a specific embodiment comprising a polypeptide or a pharmaceutically acceptable salt thereof of formula Id, X15 is T, X16 is G, and the polypeptide further comprises a linker-fatty acid. In a specific embodiment, the linker-fatty acid is 2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-εK-γE-CO-(CH2)18-CO2. In a specific embodiment comprising a polypeptide or a pharmaceutically acceptable salt thereof of formula Id, K at X35 is bound to the linker-fatty acid. In another embodiment, the polypeptide comprises SEQ ID NO: 32. In another embodiment, the polypeptide is SEQ ID NO: 32. Alternatively, the polypeptide may have at least about 90% to 97% sequence similarity to SEQ ID NO: 32. Alternatively, the polypeptide may have at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, or 97% sequence similarity to SEQ ID NO: 32.

[0067] In a specific embodiment comprising a polypeptide or a pharmaceutically acceptable salt thereof of formula Id, X15 is T, X16 is G, and the polypeptide further comprises a linker-fatty acid. In a specific embodiment, the linker-fatty acid is 2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-γE-εK-CO-(CH2)18-CO2H. In a specific embodiment comprising a polypeptide or a pharmaceutically acceptable salt thereof of formula Id, K at X35 is bound to the linker-fatty acid. In another embodiment, the polypeptide comprises SEQ ID NO: 33. In another embodiment, the polypeptide is SEQ ID NO: 33. Alternatively, the polypeptide may have at least about 90% to 97% sequence similarity to SEQ ID NO: 33. Alternatively, the polypeptide may have at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, or 97% sequence similarity to SEQ ID NO: 33.

[0068] In a specific embodiment comprising a polypeptide or a pharmaceutically acceptable salt thereof of formula Id, X15 is T, X16 is G, and the polypeptide further comprises a linker-fatty acid. In a specific embodiment, the linker-fatty acid is εK-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-γE-CO-(CH2)18-CO2H. In a specific embodiment of the polypeptide or a pharmaceutically acceptable salt thereof, K at X35 is bound to the linker-fatty acid. In another embodiment, the polypeptide comprises SEQ ID NO: 34. In another embodiment, the polypeptide is SEQ ID NO: 34. Alternatively, the polypeptide may have at least about 90% to 97% sequence similarity to SEQ ID NO: 34. Alternatively, the polypeptide may have at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, or 97% sequence similarity to SEQ ID NO: 34.

[0069] In a specific embodiment comprising a polypeptide or a pharmaceutically acceptable salt thereof of formula Id, X15 is T, X16 is G, and the polypeptide further comprises a linker-fatty acid. In a specific embodiment, the linker-fatty acid is εK-γE-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-CO-(CH2)18-CO2H. In a specific embodiment comprising a polypeptide or a pharmaceutically acceptable salt thereof of formula Id, K at X35 is bound to the linker-fatty acid. In another embodiment, the polypeptide comprises SEQ ID NO: 35. In another embodiment, the polypeptide is SEQ ID NO: 35. Alternatively, the polypeptide may have at least about 90% to 97% sequence similarity to SEQ ID NO: 35. Alternatively, the polypeptide may have at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, or 97% sequence similarity to SEQ ID NO: 35.

[0070] In a specific embodiment comprising a polypeptide or a pharmaceutically acceptable salt thereof of formula Id, X15 is T, X16 is G, and the polypeptide further comprises a linker-fatty acid. In a specific embodiment, the linker-fatty acid is γE-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-εK-CO-(CH2)18-CO2H. In a specific embodiment comprising a polypeptide or a pharmaceutically acceptable salt thereof of formula Id, K at X35 is bound to the linker-fatty acid. In another embodiment, the polypeptide comprises SEQ ID NO: 36. In another embodiment, the polypeptide is SEQ ID NO: 36. Alternatively, the polypeptide may have at least about 90% to 97% sequence similarity to SEQ ID NO: 36. Alternatively, the polypeptide may have at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, or 97% sequence similarity to SEQ ID NO: 36.

[0071] In a specific embodiment comprising a polypeptide or a pharmaceutically acceptable salt thereof of formula Id, X15 is T, X16 is G, and the polypeptide further comprises a linker-fatty acid. In a specific embodiment, the linker-fatty acid is γE-εK-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-CO-(CH2)18-CO2H. In a specific embodiment comprising a polypeptide or a pharmaceutically acceptable salt thereof of formula Id, K at X35 is bound to the linker-fatty acid. In another embodiment, the polypeptide comprises SEQ ID NO: 37. In another embodiment, the polypeptide is SEQ ID NO: 37. Alternatively, the polypeptide may have at least about 90% to 97% sequence similarity to SEQ ID NO: 37. Alternatively, the polypeptide may have at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, or 97% sequence similarity to SEQ ID NO: 37.

[0072] In a specific embodiment comprising a polypeptide or a pharmaceutically acceptable salt thereof, X15 is T, X16 is G, and the polypeptide further comprises a linker-fatty acid. In a specific embodiment, the linker-fatty acid is γE-γE-εK-CO-(CH2)18-CO2H. In a specific embodiment comprising a polypeptide or a pharmaceutically acceptable salt thereof, K at X35 is bound to the linker-fatty acid. In another embodiment, the polypeptide comprises SEQ ID NO: 38. In another embodiment, the polypeptide is SEQ ID NO: 38. Alternatively, the polypeptide may have at least about 90% to 97% sequence similarity to SEQ ID NO: 38. Alternatively, the polypeptide may have at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, or 97% sequence similarity to SEQ ID NO: 38.

[0073] In a specific embodiment comprising a polypeptide or a pharmaceutically acceptable salt thereof, X15 is T, X16 is G, and the polypeptide further comprises a linker-fatty acid. In a specific embodiment, the linker-fatty acid is γE-εK-γE-CO-(CH2)18-CO2H. In a specific embodiment comprising a polypeptide or a pharmaceutically acceptable salt thereof, K at X35 is bound to the linker-fatty acid. In another embodiment, the polypeptide comprises SEQ ID NO: 39. In another embodiment, the polypeptide is SEQ ID NO: 39. Alternatively, the polypeptide may have at least about 90% to 97% sequence similarity to SEQ ID NO: 39. Alternatively, the polypeptide may have at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, or 97% sequence similarity to SEQ ID NO: 39.

[0074] In a specific embodiment comprising a polypeptide or a pharmaceutically acceptable salt thereof of formula Id, X15 is T, X16 is G, and the polypeptide further comprises a linker-fatty acid. In a specific embodiment, the linker-fatty acid is εK-γE-γE-CO-(CH2)18-CO2H. In a specific embodiment comprising a polypeptide or a pharmaceutically acceptable salt thereof of formula Id, K at X35 is bound to the linker-fatty acid. In another embodiment, the polypeptide comprises SEQ ID NO: 40. In another embodiment, the polypeptide is SEQ ID NO: 40. Alternatively, the polypeptide may have at least about 90% to 97% sequence similarity to SEQ ID NO: 40. Alternatively, the polypeptide may have at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, or 97% sequence similarity to SEQ ID NO: 40.

[0075] In a specific embodiment comprising a polypeptide or a pharmaceutically acceptable salt thereof of formula Id, X15 is T, X16 is Aib, and the polypeptide further comprises a linker-fatty acid. In a specific embodiment, the linker-fatty acid is 2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)2-γE-CO-(CH2)18-CO2H. In a specific embodiment comprising a polypeptide or a pharmaceutically acceptable salt thereof of formula Id, K at X35 is bound to the linker-fatty acid. In another embodiment, the polypeptide comprises SEQ ID NO: 41. In another embodiment, the polypeptide is SEQ ID NO: 41. Alternatively, the polypeptide may have at least about 90% to 97% sequence similarity to SEQ ID NO: 41. Alternatively, the polypeptide may have at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, or 97% sequence similarity to SEQ ID NO: 41.

[0076] In a specific embodiment comprising a polypeptide or a pharmaceutically acceptable salt thereof of formula Id, X15 is I, X16 is Aib, and the polypeptide further comprises a linker-fatty acid moiety. In a specific embodiment, the linker-fatty acid is 2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)2-γE-CO-(CH2)18-CO2H. In a specific embodiment comprising a polypeptide or a pharmaceutically acceptable salt thereof of formula Id, K at X35 is bound to the linker-fatty acid. In another embodiment, the polypeptide comprises SEQ ID NO: 42. In another embodiment, the polypeptide is SEQ ID NO: 42. Alternatively, the polypeptide may have at least about 90% to 97% sequence similarity to SEQ ID NO: 42. Alternatively, the polypeptide may have at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, or 97% sequence similarity to SEQ ID NO: 42.

[0077] In a specific embodiment comprising a polypeptide or a pharmaceutically acceptable salt thereof, X15 is T, X16 is G, and the polypeptide further comprises a linker-fatty acid. In a specific embodiment, the linker-fatty acid is GKEKEKE-CO-(CH2)18-CO2H. In a specific embodiment comprising a polypeptide or a pharmaceutically acceptable salt thereof, K at X35 is bound to the linker-fatty acid. In another embodiment, the polypeptide comprises SEQ ID NO: 43. In another embodiment, the polypeptide is SEQ ID NO: 43. Alternatively, the polypeptide may have at least about 90% to 97% sequence similarity to SEQ ID NO: 43. Alternatively, the polypeptide may have at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, or 97% sequence similarity to SEQ ID NO: 43.

[0078] In a specific embodiment comprising a polypeptide or a pharmaceutically acceptable salt thereof of formula Id, X15 is T, X16 is G, and the polypeptide further comprises a linker-fatty acid. In a specific embodiment, the linker-fatty acid is 2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-γE2-CO-(CH2)18-CO2H. In a specific embodiment comprising a polypeptide or a pharmaceutically acceptable salt thereof of formula Id, K at X35 is bound to the linker-fatty acid. In another embodiment, the polypeptide comprises SEQ ID NO: 44. In another embodiment, the polypeptide is SEQ ID NO: 44. Alternatively, the polypeptide may have at least about 90% to 97% sequence similarity to SEQ ID NO: 44. Alternatively, the polypeptide may have at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, or 97% sequence similarity to SEQ ID NO: 44.

[0079] In a specific embodiment comprising a polypeptide or a pharmaceutically acceptable salt thereof of formula Id, X15 is I, X16 is Aib, and the polypeptide further comprises a linker-fatty acid. In a specific embodiment, the linker-fatty acid is γE2-CO-(CH2)18-CO2H. In a specific embodiment comprising a polypeptide or a pharmaceutically acceptable salt thereof of formula Id, K at X35 is bound to the linker-fatty acid. In another embodiment, the polypeptide comprises SEQ ID NO: 51. In another embodiment, the polypeptide is SEQ ID NO: 51. Alternatively, the polypeptide may have at least about 90% to 97% sequence similarity to SEQ ID NO: 51. Alternatively, the polypeptide may have at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, or 97% sequence similarity to SEQ ID NO: 51.

[0080] In another embodiment of Formula I, X1 is absent, X2 is absent, X3 is absent, X4 is P, X5 is P, X6 is P, X11 is αMeL, X13 is V or D-Val, X15 is T, X16 is Aib, X17 is αMeL, X19 is Q, X20 is K, X21 is L or Aib, X22 is L, X26 is R, X27 is A, X30 is A, X31 is R, X35 is K, X37 is A or N, and X38 is A or T. In such embodiments, the polypeptide comprises a sequence of 41 amino acids.

[0081] Therefore, in some embodiments, this document provides a polypeptide of formula Ie or a pharmaceutically acceptable salt thereof, which comprises PPPIVTS αMeL DX13PTAib αMeL LQKX21LEQERAEKARQQAKEX37X38EILAQV (SEQ ID NO: 73), where, X13 is V or D-Val [where X1 to X3 are absent, X13 is located at position 10 of the polypeptide], X21 is either L or Aib [where X21 is at position 18 of the polypeptide when X1 to X3 are absent]. X37 is either A or N [where X37 is located at position 34 of the polypeptide when X1 to X3 are absent]. X38 is either A or T [where X38 is located at position 35 of the polypeptide when X1 to X3 are absent]. The N-terminal amino acid is N-acetylated as appropriate, and the C-terminal amino acid is amylated as appropriate.

[0082] In a specific embodiment comprising a polypeptide or a pharmaceutically acceptable salt thereof of formula Ie, X13 is D-Val, X21 is Aib, X37 is N, X38 is T, and the polypeptide further comprises a linker-fatty acid. In a specific embodiment, the linker-fatty acid is γE-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-εK-CO-(CH2)18-CO2H. In a specific embodiment, K at X35 is bound to the linker-fatty acid. In some embodiments, the polypeptide comprises SEQ ID NO: 46. In some embodiments, the polypeptide is SEQ ID NO: 46. Alternatively, the polypeptide may have at least about 90% to 97% sequence similarity to SEQ ID NO: 46. Alternatively, the polypeptide may have at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, or 97% sequence similarity to SEQ ID NO: 46.

[0083] In a specific embodiment comprising a polypeptide or a pharmaceutically acceptable salt thereof of formula Id, X13 is D-Val, X21 is L, X37 is A, X38 is A, and the polypeptide further comprises a linker-fatty acid. In a specific embodiment, the linker-fatty acid is γE-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-εK-CO-(CH2)18-CO2H. In a specific embodiment comprising a polypeptide or a pharmaceutically acceptable salt thereof of formula Id, K at X35 is bound to the linker-fatty acid. In some embodiments, the polypeptide comprises SEQ ID NO: 47. In some embodiments, the polypeptide is SEQ ID NO: 47. Alternatively, the polypeptide may have at least about 90% to 97% sequence similarity to SEQ ID NO: 47. Alternatively, the polypeptide may have at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, or 97% sequence similarity to SEQ ID NO: 47.

[0084] In a particular embodiment comprising a polypeptide or a pharmaceutically acceptable salt thereof of formula Id, X13 is V, X21 is L, X37 is A, X38 is A, and the polypeptide further comprises a linker-fatty acid. In a particular embodiment, the linker-fatty acid is (γE-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-εK-CO-(CH2)18-CO2H. In a particular embodiment comprising a polypeptide or a pharmaceutically acceptable salt thereof of formula Id, K at X35 is bound to the linker-fatty acid. In some embodiments, the polypeptide comprises SEQ ID NO: 48. In some embodiments, the polypeptide is SEQ ID NO: 48. Alternatively, the polypeptide may have at least about 90% to 97% sequence similarity to SEQ ID NO: 48. Alternatively, the polypeptide may have at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, or 97% sequence similarity to SEQ ID NO: 48.

[0085] In another embodiment of Formula I, X1 is γE, X2 is absent, X3 is absent, X4 is G, X5 is P, X6 is S, X11 is αMeL, X13 is D-Val, X15 is I, X16 is Aib, X17 is αMeL, X19 is Q, X20 is I, X21 is K, X22 is L, X26 is R, X27 is A, X30 is A, X31 is R, X35 is K, X37 is N, and X38 is T. In a particular embodiment, the polypeptide further comprises a linker-fatty acid. In a particular embodiment, the linker-fatty acid is (2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)2-γE-CO-(CH2)18-CO2H. In a particular embodiment comprising the polypeptide or a pharmaceutically acceptable salt thereof, K at X35 is bound to the linker-fatty acid.

[0086] In some embodiments, the polypeptide comprises SEQ ID NO: 45. In some embodiments, the polypeptide is SEQ ID NO: 45. Alternatively, the polypeptide may have at least about 90% to 97% sequence similarity to SEQ ID NO: 45. Alternatively, the polypeptide may have at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, or 97% sequence similarity to SEQ ID NO: 45.

[0087] In another embodiment of Equation I, X1 does not exist, X2 does not exist, X3 does not exist, X4 does not exist, X5 does not exist, X6 does not exist, X11 is L, X13 is V, X15 is I, X16 is G, X17 is αMeL, X19 is Q, X20 is K, X21 is L, X22 is I, X26 is K, X27 is Q, X30 is E, X31 is K, X35 is K, X37 is N, and X38 is T.

[0088] In a particular embodiment of the polypeptide or a pharmaceutically acceptable salt thereof, the polypeptide further comprises a linker-fatty acid. In a particular embodiment, the linker-fatty acid is (2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)2γE-C20-OH)). In a particular embodiment comprising the polypeptide or a pharmaceutically acceptable salt thereof, K at X35 is bound to the linker-fatty acid.

[0089] In some embodiments, the polypeptide comprises SEQ ID NO: 49. In some embodiments, the polypeptide is SEQ ID NO: 49. Alternatively, the polypeptide may have at least about 90% to 97% sequence similarity to SEQ ID NO: 49. Alternatively, the polypeptide may have at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, or 97% sequence similarity to SEQ ID NO: 49.

[0090] In another embodiment of Equation I, X1 does not exist, X2 does not exist, X3 does not exist, X4 does not exist, X5 does not exist, X6 does not exist, X11 is L, X13 is V, X15 is I, X16 is G, X17 is αMeL, X19 is Q, X20 is K, X21 is L, X22 is L, X26 is R, X27 is A, X30 is A, X31 is R, X35 is K, X37 is A, and X38 is A.

[0091] In a particular embodiment of the polypeptide or a pharmaceutically acceptable salt thereof, the polypeptide further comprises a linker-fatty acid. In one particular embodiment, the linker-fatty acid is 2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)2-γE-CO-(CH2)18-CO2H. In a particular embodiment comprising the polypeptide or a pharmaceutically acceptable salt thereof, K at X35 is bound to the linker-fatty acid.

[0092] In some embodiments, the polypeptide comprises SEQ ID NO: 50. In other embodiments, the polypeptide is SEQ ID NO: 50. Alternatively, the polypeptide may have at least about 90% to 97% sequence similarity to SEQ ID NO: 50. Alternatively, the polypeptide may have at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, or 97% sequence similarity to SEQ ID NO: 50.

[0093] In one embodiment, this document provides a polypeptide or a pharmaceutically acceptable salt thereof comprising a sequence having at least eight amino acid modifications compared to human urocortin-2 (SEQ ID NO: 53). In some embodiments, the polypeptide provided herein comprises a sequence having at least one amino acid modification at positions 3, 19, 22, 23, 26, 30, 33, or 37 of human urocortin-2 (SEQ ID NO: 53). In some embodiments, the polypeptide provided herein comprises a sequence having at least one of the following amino acid modifications compared to human urocortin-2 (SEQ ID NO: 53): L3T, A19E, R22E, A23K, E26Q, T30E, R33E, R37Q, or any combination thereof.

[0094] In another embodiment, this document provides a polypeptide comprising any of SEQ ID NO: 1 to 52 or a pharmaceutically acceptable salt thereof. Alternatively, the polypeptide may have at least about 90% to 99% sequence similarity to any of SEQ ID NO: 1 to 52 or a pharmaceutically acceptable salt thereof. Alternatively, the polypeptide may have at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence similarity to any of SEQ ID NO: 1 to 52.

[0095] In one specific embodiment, the polypeptide comprises SEQ ID NO: 1 or a pharmaceutically acceptable salt thereof. In one specific embodiment, the polypeptide comprises SEQ ID NO: 2 or a pharmaceutically acceptable salt thereof. In one specific embodiment, the polypeptide comprises SEQ ID NO: 22 or a pharmaceutically acceptable salt thereof. In one specific embodiment, the polypeptide comprises SEQ ID NO: 25 or a pharmaceutically acceptable salt thereof. In one specific embodiment, the polypeptide comprises SEQ ID NO: 29 or a pharmaceutically acceptable salt thereof. In one specific embodiment, the polypeptide comprises SEQ ID NO: 41 or a pharmaceutically acceptable salt thereof. In one specific embodiment, the polypeptide comprises SEQ ID NO: 42 or a pharmaceutically acceptable salt thereof. In one specific embodiment, the polypeptide comprises SEQ ID NO: 45 or a pharmaceutically acceptable salt thereof. In one specific embodiment, the polypeptide comprises SEQ ID NO: 46 or a pharmaceutically acceptable salt thereof. In one specific embodiment, the polypeptide comprises SEQ ID NO: 48 or a pharmaceutically acceptable salt thereof. In one particular embodiment, the polypeptide comprises SEQ ID NO: 49 or a pharmaceutically acceptable salt thereof. In another particular embodiment, the polypeptide comprises SEQ ID NO: 50 or a pharmaceutically acceptable salt thereof.

[0096] In a particular embodiment, the polypeptide or its pharmaceutically acceptable salt system is selected from the group consisting of: SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 22, SEQ ID NO: 25, SEQ ID NO: 29, SEQ ID NO: 41, SEQ ID NO: 42, SEQ ID NO: 45, SEQ ID NO: 46, SEQ ID NO: 48, SEQ ID NO: 49, and SEQ ID NO: 50.

[0097] In another embodiment, this document provides a pharmaceutical composition comprising a polypeptide or a pharmaceutically acceptable salt thereof and at least one pharmaceutically acceptable carrier, diluent or excipient.

[0098] In another embodiment, a method is provided for treating diseases or conditions selected from the group consisting of: diabetes, obesity, chronic weight management, non-alcoholic fatty liver disease (NAFLD), non-alcoholic steatohepatitis (NASH), dyslipidemia, metabolic syndrome, chronic kidney disease (CKD), osteoarthritis (OA), obesity-related sleep apnea (OSA), sarcopenia, cachexia, sarcopenic obesity (SO), and polycystic ovarian syndrome (PCOS), the method comprising administering an effective amount of a polypeptide or a medically acceptable salt thereof to the individual in need.

[0099] In another embodiment, this document provides a polypeptide or a pharmaceutically acceptable salt thereof for use in a therapeutic.

[0100] In another embodiment, this document provides a polypeptide or a pharmaceutically acceptable salt thereof for the treatment of diseases or conditions selected from the group consisting of: diabetes, obesity, chronic weight management, non-alcoholic fatty liver disease (NAFLD), non-alcoholic steatohepatitis (NASH), dyslipidemia, metabolic syndrome, chronic kidney disease (CKD), osteoarthritis (OA), obesity-related sleep apnea (OSA), sarcopenia, cachexia, sarcopenic obesity (SO), and polycystic ovary syndrome (PCOS).

[0101] In one embodiment, the present invention provides the use of a polypeptide or a pharmaceutically acceptable salt thereof for the manufacture of an agent for treating diseases or conditions selected from the group consisting of: diabetes, obesity, chronic weight management, non-alcoholic fatty liver disease (NAFLD), non-alcoholic steatosis (NASH), dyslipidemia, metabolic syndrome, chronic kidney disease (CKD), osteoarthritis (OA), obesity-related sleep apnea (OSA), sarcopenia, cachexia, sarcopenic obesity (SO), and polycystic ovary syndrome (PCOS).

[0102] In one embodiment, this document provides a method for treating diseases or conditions selected from the group consisting of: diabetes, obesity, chronic weight management, non-alcoholic fatty liver disease (NAFLD), non-alcoholic steatosis (NASH), dyslipidemia, metabolic syndrome, chronic kidney disease (CKD), osteoarthritis (OA), obesity-related sleep apnea (OSA), sarcopenia, cachexia, sarcopenic obesity (SO), and polycystic ovary syndrome (PCOS). The method comprises administering to an individual in need an effective amount of a polypeptide or a medically acceptable salt thereof and an effective amount of additional therapeutic agent.

[0103] In one embodiment, this document provides a polypeptide or a pharmaceutically acceptable salt thereof, which can be used simultaneously, separately, or sequentially in combination with one or more additional therapeutic agents.

[0104] In one embodiment, the additional therapeutic agent is a dual agonist of the GIP and GLP-1 receptors. In a particular embodiment, the dual agonist of the GIP and GLP-1 receptors is a compound comprising SEQ ID NO: 55. In one embodiment, the additional therapeutic agent is a agonist of the GLP-1 receptor. In one embodiment, the GLP-1 receptor agonist is a compound comprising SEQ ID NO: 79, SEQ ID NO: 80, or SEQ ID NO: 81. Implementation

[0105] This application claims the benefit of U.S. Provisional Application No. 63 / 600,468, filed November 17, 2023, pursuant to 35 USC §119(e), the disclosure of which is incorporated herein by reference.

[0106] [] [Sequence List] [, , ] This application is submitted together with a sequence list in ST.26 XML format. The sequence list is provided as a file entitled "30698_WO_SL", created on October 31, 2024, and is 2,480,000 bytes in size. The ST.26 XML format sequence list information is incorporated herein by reference in its entirety.

[0107] Amino acids consist of an amino group and a carboxylic acid group. In α-amino acids (a class of 20 typical amino acids), the amino group and the carboxylic acid group are covalently attached to the same carbon, which is defined as the α-carbon. The α-carbon can also be substituted with additional functional groups, and these functional groups can have defined stereochemistry. The 20 typical amino acids are most commonly found in their L-stereoisomers, and therefore it is assumed that unless otherwise specified, reference to amino acids specifically refers to the L-stereoisomer.

[0108] Peptides are amino acid polymers formed by the covalent linkage between a carboxylic acid group on one monomer and an amino group on the next monomer. Most commonly, these are α-carboxylic acid groups and α-amino groups, and therefore unless otherwise specified, peptides written using standard single-letter or three-letter amino acid codes are assumed to be α-amino acid polymers.

[0109] Typically, when drawing amino acids, the amino group is placed on the left and the carboxylic acid group on the right. When writing a polypeptide sequence, if it is an unmodified linear α-amino acid chain, it is assumed that the first amino acid has a free amino group on the left side of the chain (H2N-polypeptide, also written as H-polypeptide or polypeptide), and that the last amino acid has a free carboxylic acid group at the rightmost end of the chain (polypeptide-COOH, usually written as polypeptide or polypeptide-OH). Therefore, the left end of the polypeptide is also called the N-terminus (attributed to the nitrogen in the amino group), and the right end of the polypeptide is also called the C-terminus (attributed to the carboxylic acid group). In the absence of additional description, if the polypeptide is written only with the standard single-letter or three-letter amino acid code, it is assumed that the polypeptide begins with a free amino group at the N-terminus and terminates with a free carboxylic acid at the C-terminus.

[0110] Peptides may further include N-terminal modifications (e.g., acetylation, Ac; methylation, Me; acetylation; or other modifications). Peptides may also have C-terminal modifications; most commonly, the C-terminal group is an acetylamine (-CONH2, usually written as peptide-NH2) rather than a carboxylic acid, but other C-terminal modifications are also known in this technique.

[0111] The amino acid sequences of the polypeptides described herein incorporate naturally occurring amino acids, which are typically encoded using standard single-letter codes (e.g., L = leucine); α-methyl-substituted residues of natural amino acids (e.g., α-methyl-leucine (αMeL); and non-natural amino acids, such as α-aminoisobutyric acid (Aib). The structures of these amino acids, as well as other monomers and modifications, are described below. N-acetyl (Ac) L-pyroglutamic acid (Pyr) 2-Aminoisobutyric acid (Aib) D-Val L-ornithine (Orn) L-2,3-Diaminopropionic acid (Dap) α-Methyl-L-leucine (αMeL) α-Methyl-L-phenylalanine (αMeF) N-Methyl-L-isoleucine (NMeI) N-Methyl-L-aspartic acid (NMeN) N-Methyl-Aspartic Acid (NMeD) β-Alanine (βA) 6-Aminohexanoic acid (Ahx) 2-[2-(2-amino-ethoxy)-ethoxy]-acetic acid (AEEA) ε-L-lysine (εK), wherein the amide bond with the monomer on the N-terminal side is represented by RCO. γL-glutamic acid (γE), wherein the amide bond with the monomer on the C-terminal side is represented by HNR. Methylene thioacetal linkage between the side chains of two Cys residues ([CXXXXC]*) Eicosanedioic acid (HO2C(CH2)18CO2H) Octadecanedioic acid (HO2C(CH2)16CO2H) Therefore, as used herein, "Ac" means N-acetylglucosinolate. As used herein, "Pyr" means L-pyroglutamic acid. As used herein, "Aib" means 2-aminoisobutyric acid. As used herein, "D-Val" means D-valine. As used herein, "Orn" means L-ornithine. As used herein, "Dap" means L-2,3-diaminopropionic acid. As used herein, "αMeL" means α-methyl-L-leucine. As used herein, "αMeF" means α-methyl-L-phenylalanine. As used herein, "NMeI" means N-methyl-L-isoleucine. As used herein, "NMeN" means N-methyl-L-aspartamine. As used herein, "NMeD" means N-methyl-L-aspartic acid. As used herein, "βA" means β-alanine. As used herein, "Ahx" means 6-aminohexanoic acid. As used herein, "AEEA" means 2-[2-(2-amino-ethoxy)-ethoxy]acetic acid. As used herein, "εK" means ε-L-lysine. As used herein, "γE" means γ-L-glutamic acid.

[0112] "N-term" can refer to N-terminus or N-terminal (when used as a modifier); "C-term" can refer to C-terminus or C-terminal (when used as a modifier).

[0113] It should be noted that the combination of beneficial features of the exemplary analogues described herein is not the result of any single modification in isolation, but is achieved through a novel combination of the structural features described herein.

[0114] In some embodiments, the present invention provides a polypeptide of formula I or a pharmaceutically acceptable salt thereof, comprising: X1X2X3X4X5X6IVTSX11DX13PX15X16X17LX19X20X21X22EQEX26X27EKX30X31QQAX35EX37X38EILAQV (SEQ ID NO: 68), where X1 is Pyr, E, γE, or does not exist. X2 is G or does not exist. X3 is G, S, or does not exist. X4 is S, P, G, or does not exist. X5 is S, P, or does not exist. X6 is G, S, P, or does not exist. X11 is L or αMeL, X13 is either V or D-Val. X15 is either T or I. X16 is either Aib or G. X17 is either L or αMeL. X19 is either Q or E. X20 is either K or I. X21 can be I, K, Aib, or L. X22 represents I or L. X26 is either R or K. X27 is either A or Q. X30 is either A or E. X31 is either R or K. X35 is either K or T. X37 is either A or N. X38 is either A or T. The N-terminal amino acid is N-acylated, N-acetylated, or N-methylated as appropriate, and the C-terminal amino acid is amylated as appropriate.

[0115] In some embodiments of Formula I, V at position 44 is the C-terminal amino acid. The C-terminus may contain amide, carboxylic acid, or other modifications. The N-terminal amino acid depends on the embodiment, but will always be the first amino acid in the sequence indicating the start of the polypeptide. Thus, in embodiments of Formula I, the N-terminal amino acid may be at positions X1, X2, X3, X4, X5, X6, or I at position 7. Furthermore, in some embodiments of Formula I, the N-terminus may contain additional modifications (e.g., acetylation, Ac; methylation, Me; acetylation; or other modifications). Additionally, if any amino acid at positions X1 to X6 is indicated as "absent," the next available amino acid will bind to the previously available amino acid. Thus, various embodiments of Formula I produce polypeptides containing different lengths, such as 38 to 44 amino acids.

[0116] In addition to the sequences described herein, the polypeptides described herein may also include one or more conserved amino acid substitutions, provided that the polypeptides are still active against CRHR2.

[0117] In some embodiments, the polypeptide or a pharmaceutically acceptable salt thereof further comprises a means for increasing the half-life of the polypeptide. In some embodiments, the polypeptide or a pharmaceutically acceptable salt thereof further comprises a means for increasing the half-life of the polypeptide via any amino acid having a functional group suitable for binding. In some embodiments, the amino acid having a functional group suitable for binding is γE, E, or K.

[0118] In some embodiments, the functional group available for binding can bind to phosphonates, tetrazolium, sulfonates, branched fatty acids, peptides, proteins (such as VHH, Fc, or monoclonal antibodies) or protein fragments (such as Fab, scFv, Fv, or scFab (single-chain Fab)). In some embodiments, the fatty acid, linker, protein, or protein fragment can further act as an albumin binder and provide the potential to generate a longer-acting compound.

[0119] In some embodiments, the tool is a fatty acid.

[0120] In some embodiments, the amino acid having the functional group for binding is K, and the binding is with the ε-amino group of K. In some embodiments, the polypeptide further comprises a fatty acid bound to K at X35, wherein the fatty acid is bound to the ε-amino group of K via a direct bond or linker between K and the fatty acid. In some embodiments, the amino acid having the functional group for binding is γE at X1, and the binding is via an amide bond between the α-amino group of γE and the carboxyl group of the fatty acid. In some embodiments, the tool for increasing the half-life of the polypeptide is a fatty acid, wherein the fatty acid binds to the polypeptide at the amino acid having the functional group available for binding via a direct bond or linker between the amino acid and the fatty acid.

[0121] In some embodiments, the N-terminal amino acid is N-acylated via a fatty acid, which is bound to the N-terminal amino acid via a direct bond or linker between the amino acid and the fatty acid. In some such embodiments, the linker-fatty acid is bound to the peptide after peptide synthesis. In other such embodiments, the linker-fatty acid is bound to the amino acid (e.g., in some embodiments, with E or γE) before being added to the peptide.

[0122] In some embodiments, the polypeptide is bound to the fatty acid via a linker between the amino acid and the fatty acid. Therefore, in some embodiments, the polypeptide is bound to the fatty acid via a direct bond between the amino acid and the fatty acid. In other embodiments, the polypeptide is bound to the fatty acid via a linker between the amino acid and the fatty acid. In some embodiments, the fatty acid is a C16-C22 fatty acid. In some embodiments, the binding of the linker to the C16-C22 fatty acid is referred to as linker-fatty acid.

[0123] Examples of saturated C16-C22 fatty acids used in this document include (but are not limited to) palmitic acid (hexadecanoic acid) (C16 monocarboxylic acid), hexadecanoic acid (C16 diacid), pearlitic acid (heptadecanoic acid) (C17 monocarboxylic acid), heptadecanoic acid (C17 diacid), stearic acid (C18 monocarboxylic acid), octadecanoic acid (C18 diacid), nonadecanoic acid (nonadecanoic acid) (C19 monocarboxylic acid), nonadecanoic acid (C19 diacid), arachidic acid (eicosanoic acid) (C20 monocarboxylic acid), eicosanoic acid (C20 diacid), icosanoic acid (icosanoic acid) (C21 monocarboxylic acid), icosanoic acid (icosanoic acid) (C22 monocarboxylic acid), and icosanoic acid (C22 diacid), including their branched chains and substituted derivatives.

[0124] In some cases, the linker may have one or more (2-[2-(2-amino-ethoxy)-ethoxy]-acetyl moieties or εK, in combination with one or more amino acids as appropriate.

[0125] Where the linker includes at least one amino acid, the amino acid may be one to five E or γE amino acid residues. In some cases, the linker may include one or two, three or four or five E or γE amino acid residues, including their D-type. For example, the linker may include one or two, three or four γE amino acid residues. Alternatively, the linker may include one to five amino acid residues (e.g., E or γE amino acids) in combination with one to five (2-[2-(2-amino-ethoxy)-ethoxy]-acetyl) ("AEEA") or one to five εK moieties. Specifically, the linker may be a combination of one to five E or γE amino acids with one to five (2-[2-(2-amino-ethoxy)-ethoxy]-acetyl) ("AEEA") or one to five εK moieties, or a combination of one to five Glu or γGlu amino acids with one to five εK moieties. In some cases, the linker may be a combination of one, two, or three γGlu amino acids with one or two (2-[2-(2-amino-ethoxy)-ethoxy]-acetyl) moieties or eK moieties.

[0126] In certain embodiments, the polypeptide or a pharmaceutically acceptable salt thereof further comprises a linker-fatty acid, wherein the linker-fatty acid is selected from the group consisting of: (2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-PKE-γE-CO-(CH2)18-CO2H, Ahx-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-γE-CO-(CH2)18-CO2H, γE-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-Ahx-CO-(CH2)18-CO2H, γE-GKEKEKE-CO-(CH2)18-CO2H, G-(βA)3- γE-CO-(CH2)18-CO2H, G-(βA)4-γE-CO-(CH2)18-CO2H, (2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-KEKEKE-γE-CO-(CH2)18-CO2H, βA-S-βA-S-γE-CO-(CH2)18-CO2H, γE-PKE-Ahx-CO-(CH2)18-CO2H, γE-PKE-εK-CO-(CH2)18-CO2H, γE-PPP-εK-CO-(CH2)18-CO2H, γE-PPPPPP-CO-(CH2)18-CO2H, γE-PEP EPE-CO-(CH2)18-CO2H, γE-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)2-εK-CO-(CH2)18-CO2H, γE-εK-γE-εK-CO-(CH2)18-CO2H, γE-εK-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-εK-CO-(CH2)18-CO2H, γE-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-εK-CO-(CH2)16-CO2H, 2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-εK-CO-(CH2)18- CO2H, (2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-K-CO-(CH2)18-CO2H, (2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-εK-CO-(CH2)16-CO2H, (2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-Dap-CO-(CH2)18-CO2H, (2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)2-γE-CO-(CH2)18-CO2H, γE-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-γE-CO-(CH2)18-CO2H,γE2-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-CO-(CH2)18-CO2H, 2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-εK-γE-CO-(CH2)18-CO2, 2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-γE-εK-CO-(CH2)18-CO2H, εK-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-γE-CO-(CH2)18-CO2H, εK-γE-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-γE-CO-(CH2)18-CO2H, εK-γE-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)- γE-εK-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-CO-(CH2)18-CO2H, γE-γE-εK-CO-(CH2)18-CO2H, γE-εK-γE-CO-(CH2)18-CO2H, εK-γE-γE-CO-(CH2)18-CO2H, K(GKEKEKE-CO-(CH2)18-CO2H), 2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-γE2-CO-(CH2)18-CO2H, γE- (2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-εK-CO-(CH2)18-CO2H, 2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)2γE-C20-OH, and γE-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-εK-CO-(CH2)18-CO2H.

[0127] In some embodiments, the N-terminal amino acid is N-acetylated. In some embodiments, the N-terminal amino acid is N-methylated. In some embodiments, the C-terminal amino acid is methylated.

[0128] In one embodiment of Formula I, X1 is absent, X2 is absent, X3 is G, X4 is S, X5 is P, X6 is S, X11 is αMeL, X13 is V, X15 is T, X16 is Aib, X17 is αMeL, X19 is Q, X20 is K, X21 is I or L, X22 is L, X26 is R, X27 is A, X30 is A, X31 is R, X35 is K, X37 is A, and X38 is A. In this type of embodiment, the polypeptide comprises a sequence of 42 amino acids.

[0129] Therefore, in some embodiments, this document provides a polypeptide of formula Ia or a pharmaceutically acceptable salt thereof, comprising:

[0130] GSPSIVTS αMeL DVPT Aib αMeL LQKX21LEQERAEKARQQAKEAAEILAQV (SEQ ID NO: 69), where X21 is either I or L [where X21 is at position 19 of the polypeptide when X1 to X2 are absent]. The N-terminal amino acid is N-acetylated as appropriate, and the C-terminal amino acid is amylated as appropriate.

[0131] In some embodiments of Formula Ia, X21 is I. In some embodiments, the polypeptide comprises SEQ ID NO: 20. In another embodiment, the polypeptide is SEQ ID NO: 20. Alternatively, the polypeptide may have at least about 90% to 97% sequence similarity to SEQ ID NO: 20. Alternatively, the polypeptide may have at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, or 97% sequence similarity to SEQ ID NO: 20. Exemplary embodiments comprising SEQ ID NO: 20 are shown.

[0132] In a specific embodiment of formula Ia comprising a polypeptide or a pharmaceutically acceptable salt thereof, X21 is I and the polypeptide further comprises a linker-fatty acid. In a specific embodiment, the linker-fatty acid is γE-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-εK-CO-(CH2)18-CO2H. In a specific embodiment of formula Ia comprising a polypeptide or a pharmaceutically acceptable salt thereof, K at X35 is bound to the linker-fatty acid. In another embodiment, the polypeptide comprises SEQ ID NO: 1. In another embodiment, the polypeptide is SEQ ID NO: 1. Alternatively, the polypeptide may have at least about 90% to 97% sequence similarity to SEQ ID NO: 1. Alternatively, the polypeptide may have at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, or 97% sequence similarity to SEQ ID NO: 1. An exemplary embodiment comprising SEQ ID NO: 1 is shown.

[0133] In a specific embodiment of Formula Ia comprising a polypeptide or a pharmaceutically acceptable salt thereof, X21 is L and the polypeptide further comprises a linker-fatty acid. In a specific embodiment, the linker-fatty acid is γE-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-εK-CO-(CH2)18-CO2H. In a specific embodiment of Formula Ia comprising a polypeptide or a pharmaceutically acceptable salt thereof, K at X35 is bound to the linker-fatty acid. In another embodiment, the polypeptide comprises SEQ ID NO: 2. In another embodiment, the polypeptide is SEQ ID NO: 2. Alternatively, the polypeptide may have at least about 90% to 97% sequence similarity to SEQ ID NO: 2. Alternatively, the polypeptide may have at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, or 97% sequence similarity to SEQ ID NO: 2. An exemplary embodiment comprising SEQ ID NO: 2 is shown.

[0134] In a specific embodiment of formula Ia comprising a polypeptide or a pharmaceutically acceptable salt thereof, X21 is I and the polypeptide further comprises a linker-fatty acid. In a specific embodiment, the linker-fatty acid is 2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-PKE-γE-CO-(CH2)18-CO2H. In a specific embodiment of formula Ia comprising a polypeptide or a pharmaceutically acceptable salt thereof, K at X35 is bound to the linker-fatty acid. In another embodiment, the polypeptide comprises SEQ ID NO: 3. In another embodiment, the polypeptide is SEQ ID NO: 3. Alternatively, the polypeptide may have at least about 90% to 97% sequence similarity to SEQ ID NO: 3. Alternatively, the polypeptide may have at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, or 97% sequence similarity to SEQ ID NO: 3. An exemplary embodiment comprising SEQ ID NO: 3 is shown.

[0135] In a specific embodiment of formula Ia comprising a polypeptide or a pharmaceutically acceptable salt thereof, X21 is I and the polypeptide further comprises a linker-fatty acid. In a specific embodiment, the linker-fatty acid is Ahx-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-γE-CO-(CH2)18-CO2H. In a specific embodiment of formula Ia comprising a polypeptide or a pharmaceutically acceptable salt thereof, K at X35 is bound to the linker-fatty acid. In another embodiment, the polypeptide comprises SEQ ID NO: 4. In another embodiment, the polypeptide is SEQ ID NO: 4. Alternatively, the polypeptide may have at least about 90% to 97% sequence similarity to SEQ ID NO: 4. Alternatively, the polypeptide may have at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, or 97% sequence similarity to SEQ ID NO: 4. An exemplary embodiment comprising SEQ ID NO: 4 is shown.

[0136] In a specific embodiment of formula Ia comprising a polypeptide or a pharmaceutically acceptable salt thereof, X21 is I and the polypeptide further comprises a linker-fatty acid. In a specific embodiment, the linker-fatty acid is γE-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-Ahx-CO-(CH2)18-CO2H. In a specific embodiment of formula Ia comprising a polypeptide or a pharmaceutically acceptable salt thereof, K at X35 is bound to the linker-fatty acid. In another embodiment, the polypeptide comprises SEQ ID NO: 5. In another embodiment, the polypeptide is SEQ ID NO: 5. Alternatively, the polypeptide may have at least about 90% to 97% sequence similarity to SEQ ID NO: 5. Alternatively, the polypeptide may have at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, or 97% sequence similarity to SEQ ID NO: 5. An exemplary embodiment comprising SEQ ID NO: 5 is shown.

[0137] In a specific embodiment comprising a polypeptide or a pharmaceutically acceptable salt thereof of formula Ia, X21 is I and the polypeptide further comprises a linker-fatty acid. In a specific embodiment, the linker-fatty acid is γE-GKEKEKE-CO-(CH2)18-CO2H. In a specific embodiment comprising a polypeptide or a pharmaceutically acceptable salt thereof of formula Ia, K at X35 is bound to the linker-fatty acid. In another embodiment, the polypeptide comprises SEQ ID NO: 6. In another embodiment, the polypeptide is SEQ ID NO: 6. Alternatively, the polypeptide may have at least about 90% to 97% sequence similarity to SEQ ID NO: 6. Alternatively, the polypeptide may have at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, or 97% sequence similarity to SEQ ID NO: 6. An exemplary embodiment comprising SEQ ID NO: 6 is shown.

[0138] In a specific embodiment comprising a polypeptide or a pharmaceutically acceptable salt thereof of formula Ia, X21 is I and the polypeptide further comprises a linker-fatty acid. In a specific embodiment, the linker-fatty acid is G-(βA)3-γE-CO-(CH2)18-CO2H. In a specific embodiment comprising a polypeptide or a pharmaceutically acceptable salt thereof of formula Ia, K at X35 is bound to the linker-fatty acid. In another embodiment, the polypeptide comprises SEQ ID NO: 7. In another embodiment, the polypeptide is SEQ ID NO: 7. Alternatively, the polypeptide may have at least about 90% to 97% sequence similarity to SEQ ID NO: 7. Alternatively, the polypeptide may have at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, or 97% sequence similarity to SEQ ID NO: 7. An exemplary embodiment comprising SEQ ID NO: 7 is shown.

[0139] In a specific embodiment comprising a polypeptide or a pharmaceutically acceptable salt thereof of formula Ia, X21 is I and the polypeptide further comprises a linker-fatty acid. In a specific embodiment, the linker-fatty acid is G-(βA)4-γE-CO-(CH2)18-CO2H. In a specific embodiment comprising a polypeptide or a pharmaceutically acceptable salt thereof of formula Ia, K at X35 is bound to the linker-fatty acid. In another embodiment, the polypeptide comprises SEQ ID NO: 8. In another embodiment, the polypeptide is SEQ ID NO: 8. Alternatively, the polypeptide may have at least about 90% to 97% sequence similarity to SEQ ID NO: 8. Alternatively, the polypeptide may have at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, or 97% sequence similarity to SEQ ID NO: 8. An exemplary embodiment comprising SEQ ID NO: 8 is shown.

[0140] In a specific embodiment of formula Ia comprising a polypeptide or a pharmaceutically acceptable salt thereof, X21 is I and the polypeptide further comprises a linker-fatty acid. In a specific embodiment, the linker-fatty acid is 2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-KEKEKE-γE-CO-(CH2)18-CO2H. In a specific embodiment of formula Ia comprising a polypeptide or a pharmaceutically acceptable salt thereof, K at X35 is bound to the linker-fatty acid. In another embodiment, the polypeptide comprises SEQ ID NO: 9. In another embodiment, the polypeptide is SEQ ID NO: 9. Alternatively, the polypeptide may have at least about 90% to 97% sequence similarity to SEQ ID NO: 9. Alternatively, the polypeptide may have at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, or 97% sequence similarity to SEQ ID NO: 9. An exemplary embodiment comprising SEQ ID NO: 9 is shown.

[0141] In a specific embodiment comprising a polypeptide or a pharmaceutically acceptable salt thereof of formula Ia, X21 is I and the polypeptide further comprises a linker-fatty acid. In a specific embodiment, the linker-fatty acid is βA-S-βA-S-γE-CO-(CH2)18-CO2H. In a specific embodiment comprising a polypeptide or a pharmaceutically acceptable salt thereof of formula Ia, K at X35 is bound to the linker-fatty acid. In another embodiment, the polypeptide comprises SEQ ID NO: 10. In another embodiment, the polypeptide is SEQ ID NO: 10. Alternatively, the polypeptide may have at least about 90% to 97% sequence similarity to SEQ ID NO: 10. Alternatively, the polypeptide may have at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, or 97% sequence similarity to SEQ ID NO: 10. An exemplary embodiment comprising SEQ ID NO: 10 is shown.

[0142] In a specific embodiment comprising a polypeptide or a pharmaceutically acceptable salt thereof of formula Ia, X21 is I and the polypeptide further comprises a linker-fatty acid. In a specific embodiment, the linker-fatty acid is γE-PKE-Ahx-CO-(CH2)18-CO2H. In a specific embodiment comprising a polypeptide or a pharmaceutically acceptable salt thereof of formula Ia, K at X35 is bound to the linker-fatty acid. In another embodiment, the polypeptide comprises SEQ ID NO: 11. In another embodiment, the polypeptide is SEQ ID NO: 11. Alternatively, the polypeptide may have at least about 90% to 97% sequence similarity to SEQ ID NO: 11. Alternatively, the polypeptide may have at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, or 97% sequence similarity to SEQ ID NO: 11. An exemplary embodiment comprising SEQ ID NO: 11 is shown.

[0143] In a specific embodiment of Formula Ia comprising a polypeptide or a pharmaceutically acceptable salt thereof, X21 is L and the polypeptide further comprises a linker-fatty acid. In a specific embodiment, the linker-fatty acid is γE-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-εK-CO-(CH2)16-CO2H. In a specific embodiment of Formula Ia comprising a polypeptide or a pharmaceutically acceptable salt thereof, K at X35 is bound to the linker-fatty acid. In another embodiment, the polypeptide comprises SEQ ID NO: 12. In another embodiment, the polypeptide is SEQ ID NO: 12. Alternatively, the polypeptide may have at least about 90% to 97% sequence similarity to SEQ ID NO: 12. Alternatively, the polypeptide may have at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, or 97% sequence similarity to SEQ ID NO: 12. An exemplary embodiment comprising SEQ ID NO: 12 is shown.

[0144] In a specific embodiment comprising a polypeptide or a pharmaceutically acceptable salt thereof of formula Ia, X21 is I and the polypeptide further comprises a linker-fatty acid. In a specific embodiment, the linker-fatty acid is γE-PKE-εK-CO-(CH2)18-CO2H. In a specific embodiment comprising a polypeptide or a pharmaceutically acceptable salt thereof of formula Ia, K at X35 is bound to the linker-fatty acid. Alternatively, the polypeptide may have at least about 90% to 97% sequence similarity to SEQ ID NO: 13. Alternatively, the polypeptide may have at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, or 97% sequence similarity to SEQ ID NO: 13. An exemplary embodiment comprising SEQ ID NO: 13 is shown.

[0145] In a specific embodiment comprising a polypeptide or a pharmaceutically acceptable salt thereof of formula Ia, X21 is I and the polypeptide further comprises a linker-fatty acid. In a specific embodiment, the linker-fatty acid is γE-PPP-εK-CO-(CH2)18-CO2H. In a specific embodiment comprising a polypeptide or a pharmaceutically acceptable salt thereof of formula Ia, K at X35 is bound to the linker-fatty acid. In another embodiment, the polypeptide comprises SEQ ID NO: 14. In another embodiment, the polypeptide is SEQ ID NO: 14. Alternatively, the polypeptide may have at least about 90% to 97% sequence similarity to SEQ ID NO: 14. Alternatively, the polypeptide may have at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, or 97% sequence similarity to SEQ ID NO: 14. An exemplary embodiment comprising SEQ ID NO: 14 is shown.

[0146] In a specific embodiment comprising a polypeptide or a pharmaceutically acceptable salt thereof of formula Ia, X21 is I and the polypeptide further comprises a linker-fatty acid. In a specific embodiment, the linker-fatty acid is γE-PPPPPP-CO-(CH2)18-CO2H. In a specific embodiment comprising a polypeptide or a pharmaceutically acceptable salt thereof of formula Ia, K at X35 is bound to the linker-fatty acid. In another embodiment, the polypeptide comprises SEQ ID NO: 15. In another embodiment, the polypeptide is SEQ ID NO: 15. Alternatively, the polypeptide may have at least about 90% to 97% sequence similarity to SEQ ID NO: 15. Alternatively, the polypeptide may have at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, or 97% sequence similarity to SEQ ID NO: 15. An exemplary embodiment comprising SEQ ID NO: 15 is shown.

[0147] In a specific embodiment comprising a polypeptide or a pharmaceutically acceptable salt thereof of formula Ia, X21 is I and the polypeptide further comprises a linker-fatty acid. In a specific embodiment, the linker-fatty acid is γE-PEPEPE-CO-(CH2)18-CO2H. In a specific embodiment comprising a polypeptide or a pharmaceutically acceptable salt thereof of formula Ia, K at X35 is bound to the linker-fatty acid. In another embodiment, the polypeptide comprises SEQ ID NO: 16. In another embodiment, the polypeptide is SEQ ID NO: 16. Alternatively, the polypeptide may have at least about 90% to 97% sequence similarity to SEQ ID NO: 16. Alternatively, the polypeptide may have at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, or 97% sequence similarity to SEQ ID NO: 16. An exemplary embodiment comprising SEQ ID NO: 16 is shown.

[0148] In a specific embodiment of formula Ia comprising a polypeptide or a pharmaceutically acceptable salt thereof, X21 is I and the polypeptide further comprises a linker-fatty acid. In a specific embodiment, the linker-fatty acid is γE-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)2-εK-CO-(CH2)18-CO2H. In a specific embodiment of formula Ia comprising a polypeptide or a pharmaceutically acceptable salt thereof, K at X35 is bound to the linker-fatty acid. In another embodiment, the polypeptide comprises SEQ ID NO: 17. In another embodiment, the polypeptide is SEQ ID NO: 17. Alternatively, the polypeptide may have at least about 90% to 97% sequence similarity to SEQ ID NO: 17. Alternatively, the polypeptide may have at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, or 97% sequence similarity to SEQ ID NO: 17. An exemplary embodiment comprising SEQ ID NO: 17 is shown.

[0149] In a specific embodiment comprising a polypeptide or a pharmaceutically acceptable salt thereof of formula Ia, X21 is I and the polypeptide further comprises a linker-fatty acid. In a specific embodiment, the linker-fatty acid is γE-εK-γE-εK-CO-(CH2)18-CO2H. In a specific embodiment comprising a polypeptide or a pharmaceutically acceptable salt thereof of formula Ia, K at X35 is bound to the linker-fatty acid. Alternatively, the polypeptide may have at least about 90% to 99% sequence similarity to SEQ ID NO: 18. Alternatively, the polypeptide may have at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, or 97% sequence similarity to SEQ ID NO: 18. An exemplary embodiment comprising SEQ ID NO: 18 is shown.

[0150] In a specific embodiment of formula Ia comprising a polypeptide or a pharmaceutically acceptable salt thereof, X21 is I and the polypeptide further comprises a linker-fatty acid. In a specific embodiment, the linker-fatty acid is γE-εK-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-εK-CO-(CH2)18-CO2H. In a specific embodiment of formula Ia comprising a polypeptide or a pharmaceutically acceptable salt thereof, K at X35 is bound to the linker-fatty acid. In another embodiment, the polypeptide comprises SEQ ID NO: 19. In another embodiment, the polypeptide is SEQ ID NO: 19. Alternatively, the polypeptide may have at least about 90% to 97% sequence similarity to SEQ ID NO: 19. Alternatively, the polypeptide may have at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, or 97% sequence similarity to SEQ ID NO: 19. An exemplary embodiment comprising SEQ ID NO: 19 is shown.

[0151] In a specific embodiment of formula Ia comprising a polypeptide or a pharmaceutically acceptable salt thereof, X21 is I and the polypeptide further comprises a linker-fatty acid. In a specific embodiment, the linker-fatty acid is γE-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-εK-CO-(CH2)16-CO2H. In a specific embodiment of formula Ia comprising a polypeptide or a pharmaceutically acceptable salt thereof, K at X35 is bound to the linker-fatty acid. In another embodiment, the polypeptide comprises SEQ ID NO: 21. In another embodiment, the polypeptide is SEQ ID NO: 21. Alternatively, the polypeptide may have at least about 90% to 97% sequence similarity to SEQ ID NO: 21. Alternatively, the polypeptide may have at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, or 97% sequence similarity to SEQ ID NO: 21. An exemplary embodiment comprising SEQ ID NO: 21 is shown.

[0152] In another embodiment of Formula I, X1 is Pyr, X2 is G, X3 is S, X4 is P, X5 is S, X6 is G, X11 is αMeL, X13 is V, X15 is T, X16 is Aib, X17 is αMeL, X19 is Q, X20 is K, X21 is I, X22 is L, X26 is R, X27 is A, X30 is A, X31 is R, X35 is K, X37 is A, and X38 is A. In such embodiments, the polypeptide comprises a sequence of 44 amino acids.

[0153] Therefore, in some embodiments, the present invention provides a polypeptide of formula 1b or a pharmaceutically acceptable salt thereof, comprising:

[0154] Pyr GSPSGIVTS αMeL DVPT Aib αMeL LQKILEQERAEKARQQAKEAAEILAQV (SEQ ID NO: 70) or a medically acceptable salt thereof,

[0155] The C-terminal amino acid is acetaminated as appropriate.

[0156] In another embodiment, the polypeptide comprises SEQ ID NO: 23. In another embodiment, the polypeptide is SEQ ID NO: 23. Alternatively, the polypeptide may have at least about 90% to 97% sequence similarity to SEQ ID NO: 23. Alternatively, the polypeptide may have at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, or 97% sequence similarity to SEQ ID NO: 23. Exemplary embodiments comprising SEQ ID NO: 23 are shown.

[0157] In a specific embodiment comprising a polypeptide or a pharmaceutically acceptable salt thereof of formula Ib, the polypeptide further comprises a linker-fatty acid. In a specific embodiment, the linker-fatty acid is γE-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-εK-CO-(CH2)18-CO2H. In a specific embodiment comprising a polypeptide or a pharmaceutically acceptable salt thereof of formula Ib, K at X35 is bound to the linker-fatty acid. In another embodiment, the polypeptide comprises SEQ ID NO: 22. In another embodiment, the polypeptide is SEQ ID NO: 22. Alternatively, the polypeptide may have at least about 90% to 97% sequence similarity to SEQ ID NO: 22. Alternatively, the polypeptide may have at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, or 97% sequence similarity to SEQ ID NO: 22. An exemplary embodiment comprising SEQ ID NO: 22 is shown.

[0158] In a specific embodiment comprising a polypeptide or a pharmaceutically acceptable salt thereof of formula Ib, the polypeptide further comprises a linker-fatty acid. In a specific embodiment, the linker-fatty acid is γE-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-εK-CO-(CH2)16-CO2H. In a specific embodiment comprising a polypeptide or a pharmaceutically acceptable salt thereof of formula Ib, K at X35 is bound to the linker-fatty acid. In another embodiment, the polypeptide comprises SEQ ID NO: 24. In another embodiment, the polypeptide is SEQ ID NO: 24. Alternatively, the polypeptide may have at least about 90% to 97% sequence similarity to SEQ ID NO: 24. Alternatively, the polypeptide may have at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, or 97% sequence similarity to SEQ ID NO: 24. An exemplary embodiment comprising SEQ ID NO: 24 is shown.

[0159] In another embodiment of Equation I, X1 is γE, E, or does not exist; X2 does not exist; X3 is G; X4 is S; X5 is P; X6 is S; X11 is αMeL; X13 is V; X15 is T; X16 is Aib; X17 is αMeL; X19 is Q or E; X20 is K; X21 is I; X22 is L; X26 is R; X27 is A; X30 is A; X31 is R; X35 is T; X37 is A; and X38 is A.

[0160] In another embodiment of Formula I, X1 is absent, X2 is absent, X3 is G, X4 is S, X5 is P, X6 is S, X11 is αMeL, X13 is V, X15 is T, X16 is Aib, X17 is αMeL, X19 is Q, X20 is K, X21 is I, X22 is L, X26 is R, X27 is A, X30 is A, X31 is R, X35 is T, X37 is A, and X38 is A. In such embodiments, the polypeptide comprises a sequence of 42 amino acids.

[0161] In another embodiment, the polypeptide comprises SEQ ID NO: 27. In another embodiment, the polypeptide is SEQ ID NO: 27. Alternatively, the polypeptide may have at least about 90% to 97% sequence similarity to SEQ ID NO: 27. Alternatively, the polypeptide may have at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, or 97% sequence similarity to SEQ ID NO: 27. Exemplary embodiments comprising SEQ ID NO: 27 are shown.

[0162] In another embodiment of Formula I, X1 is γE or E, X2 is absent, X3 is G, X4 is S, X5 is P, X6 is S, X11 is αMeL, X13 is V, X15 is T, X16 is Aib, X17 is αMeL, X19 is Q or E, X20 is K, X21 is I, X22 is L, X26 is R, X27 is A, X30 is A, X31 is R, X35 is T, X37 is A, and X38 is A. In such embodiments, the polypeptide comprises a sequence of 43 amino acids.

[0163] In a particular embodiment comprising a polypeptide or a pharmaceutically acceptable salt thereof of formula Ic, X1 is γE and the polypeptide further comprises a linker-fatty acid. In a particular embodiment, the polypeptide further comprises a fatty acid bonded to γE at position X1 via an amide bond between the α-amino group of γE and the carboxylic acid of the linker-fatty acid monomer of (2-{2-(2-amino-ethoxy)-ethoxy]acetic acid.

[0164] Therefore, in some embodiments, the present invention provides a polypeptide of formula Ic or a pharmaceutically acceptable salt thereof, comprising: X1GSPSIVTS αMeL DVPT Aib αMeLLX19KILEQERAEKARQQATEAAEILAQV (SEQ ID NO: 71), where X1 is γE or E, X19 is either Q or E [where X19 is at position 18 of the polypeptide when X2 is absent], The C-terminal amino acid is acetaminated as appropriate.

[0165] In a particular embodiment of the polypeptide or its pharmaceutically acceptable salt, X1 is γE, which is bound to X3 at G via an amide bond between the γ-carboxylic acid group of γE and the amino group of G.

[0166] In a particular embodiment comprising a polypeptide or a pharmaceutically acceptable salt thereof of formula Ic, X1 is γE, X19 is Q, and the linker-fatty acid is (2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-εK-CO-(CH2)18-CO2H. In another embodiment, the polypeptide comprises SEQ ID NO: 25. In another embodiment, the polypeptide is SEQ ID NO: 25. Alternatively, the polypeptide may have at least about 90% to 97% sequence similarity to SEQ ID NO: 25. Alternatively, the polypeptide may have at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, or 97% sequence similarity to SEQ ID NO: 25. Exemplary embodiments comprising SEQ ID NO: 25 are shown.

[0167] In a particular embodiment comprising a polypeptide or a pharmaceutically acceptable salt thereof of formula Ic, X1 is γE, X19 is Q, and the linker-fatty acid is (2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-K-CO-(CH2)18-CO2H. In another embodiment, the polypeptide comprises SEQ ID NO: 26. In another embodiment, the polypeptide is SEQ ID NO: 26. Alternatively, the polypeptide may have at least about 90% to 97% sequence similarity to SEQ ID NO: 26. Alternatively, the polypeptide may have at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, or 97% sequence similarity to SEQ ID NO: 26. Exemplary embodiments comprising SEQ ID NO: 26 are shown.

[0168] In a particular embodiment comprising a polypeptide or a pharmaceutically acceptable salt thereof of formula Ic, X1 is γE, X19 is Q, and the linker-fatty acid is (2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-εK-CO-(CH2)16-CO2H. In another embodiment, the polypeptide comprises SEQ ID NO: 28. In another embodiment, the polypeptide is SEQ ID NO: 28. Alternatively, the polypeptide may have at least about 90% to 97% sequence similarity to SEQ ID NO: 28. Alternatively, the polypeptide may have at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, or 97% sequence similarity to SEQ ID NO: 28. Exemplary embodiments comprising SEQ ID NO: 28 are shown.

[0169] In a particular embodiment comprising a polypeptide or a pharmaceutically acceptable salt thereof of formula Ic, X1 is γE, X19 is E, and the linker-fatty acid is (2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-Dap-CO-(CH2)18-CO2H. In another embodiment, the polypeptide comprises SEQ ID NO: 52. In another embodiment, the polypeptide is SEQ ID NO: 52. Alternatively, the polypeptide may have at least about 90% to 97% sequence similarity to SEQ ID NO: 52. Alternatively, the polypeptide may have at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, or 97% sequence similarity to SEQ ID NO: 52. Exemplary embodiments comprising SEQ ID NO: 52 are shown.

[0170] In another embodiment of Formula I, X1 is absent, X2 is absent, X3 is absent, X4 is absent, X5 is absent, X6 is absent, X11 is αMeL, X13 is D-Val, X15 is T or I, X16 is Aib or G, X17 is αMeL, X19 is Q, X20 is I, X21 is K, X22 is L, X26 is R, X27 is A, X30 is A, X31 is R, X35 is K, X37 is N, and X38 is T. In such embodiments, the polypeptide comprises a sequence of 38 amino acids.

[0171] In some embodiments, the present invention provides a compound of formula Id or a pharmaceutically acceptable salt thereof, comprising... IVTS αMeL D D-Val PX15X16αMeLLQIKLEQERAEKARQQAKENTEILAQV (SEQ ID NO: 72), where X15 is either T or I [where X1 to X6 are absent, X15 is located at position 9 of the polypeptide], X16 is either G or Aib [where X1 to X6 are absent, X16 is at position 10 of the polypeptide], The N-terminal amino acid is N-acetylated as appropriate, and the C-terminal amino acid is amylated as appropriate.

[0172] In a particular embodiment comprising a polypeptide or a pharmaceutically acceptable salt thereof, X15 is T, X16 is G, and the polypeptide further comprises a linker-fatty acid. In a particular embodiment, the linker-fatty acid is 2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)2-γE-CO-(CH2)18-CO2H. In a particular embodiment comprising a polypeptide or a pharmaceutically acceptable salt thereof, K at X35 is bound to the linker-fatty acid. In another embodiment, the polypeptide is SEQ ID NO: 29. Alternatively, the polypeptide may have at least about 90% to 97% sequence similarity to SEQ ID NO: 29. Alternatively, the polypeptide may have at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, or 97% sequence similarity to SEQ ID NO: 29. An exemplary embodiment comprising SEQ ID NO: 29 is shown.

[0173] In a specific embodiment comprising a polypeptide or a pharmaceutically acceptable salt thereof of formula Id, X15 is T, X16 is G, and the polypeptide further comprises a linker-fatty acid. In a specific embodiment, the linker-fatty acid is γE-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-γE-CO-(CH2)18-CO2H. In a specific embodiment comprising a polypeptide or a pharmaceutically acceptable salt thereof of formula Id, K at X35 is bound to the linker-fatty acid. In another embodiment, the polypeptide comprises SEQ ID NO: 30. In another embodiment, the polypeptide is SEQ ID NO: 30. Alternatively, the polypeptide may have at least about 90% to 97% sequence similarity to SEQ ID NO: 30. Alternatively, the polypeptide may have at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, or 97% sequence similarity to SEQ ID NO: 30. An exemplary embodiment comprising SEQ ID NO: 30 is shown.

[0174] In a specific embodiment comprising a polypeptide or a pharmaceutically acceptable salt thereof of formula Id, X15 is T, X16 is G, and the polypeptide further comprises a linker-fatty acid. In a specific embodiment, the linker-fatty acid is γE2-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-CO-(CH2)18-CO2H. In a specific embodiment comprising a polypeptide or a pharmaceutically acceptable salt thereof of formula Id, K at X35 is bound to the linker-fatty acid. In another embodiment, the polypeptide comprises SEQ ID NO: 31. In another embodiment, the polypeptide is SEQ ID NO: 31. Alternatively, the polypeptide may have at least about 90% to 97% sequence similarity to SEQ ID NO: 31. Alternatively, the polypeptide may have at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, or 97% sequence similarity to SEQ ID NO: 31. An exemplary embodiment comprising SEQ ID NO: 31 is shown.

[0175] In a particular embodiment comprising a polypeptide or a pharmaceutically acceptable salt thereof, X15 is T, X16 is G, and the polypeptide further comprises a linker-fatty acid. In a particular embodiment, the linker-fatty acid is 2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-εK-γE-CO-(CH2)18-CO2. In a particular embodiment comprising a polypeptide or a pharmaceutically acceptable salt thereof, K at X35 is bound to the linker-fatty acid. In another embodiment, the polypeptide comprises SEQ ID NO: 32. In another embodiment, the polypeptide is SEQ ID NO: 32. Alternatively, the polypeptide may have at least about 90% to 97% sequence similarity to SEQ ID NO: 32. Alternatively, the polypeptide may have at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, or 97% sequence similarity to SEQ ID NO: 32. An exemplary embodiment comprising SEQ ID NO: 32 is shown.

[0176] In a particular embodiment comprising a polypeptide or a pharmaceutically acceptable salt thereof of formula Id, X15 is T, X16 is G, and the polypeptide further comprises a linker-fatty acid. In a particular embodiment, the linker-fatty acid is 2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-γE-εK-CO-(CH2)18-CO2H. In a particular embodiment comprising a polypeptide or a pharmaceutically acceptable salt thereof of formula Id, K at X35 is bound to the linker-fatty acid. In another embodiment, the polypeptide comprises SEQ ID NO: 33. In another embodiment, the polypeptide is SEQ ID NO: 33. Alternatively, the polypeptide may have at least about 90% to 97% sequence similarity to SEQ ID NO: 33. Alternatively, the polypeptide may have at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, or 97% sequence similarity to SEQ ID NO: 33. An exemplary embodiment comprising SEQ ID NO: 33 is shown.

[0177] In a particular embodiment comprising a polypeptide or a pharmaceutically acceptable salt thereof of formula Id, X15 is T, X16 is G, and the polypeptide further comprises a linker-fatty acid. In a particular embodiment, the linker-fatty acid is εK-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-γE-CO-(CH2)18-CO2H. In a particular embodiment of the polypeptide or a pharmaceutically acceptable salt thereof, K at X35 is bound to the linker-fatty acid. In another embodiment, the polypeptide comprises SEQ ID NO: 34. In another embodiment, the polypeptide is SEQ ID NO: 34. Alternatively, the polypeptide may have at least about 90% to 97% sequence similarity to SEQ ID NO: 34. Alternatively, the polypeptide may have at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, or 97% sequence similarity to SEQ ID NO: 34. An exemplary embodiment comprising SEQ ID NO: 34 is shown.

[0178] In a specific embodiment comprising a polypeptide or a pharmaceutically acceptable salt thereof of formula Id, X15 is T, X16 is G, and the polypeptide further comprises a linker-fatty acid. In a specific embodiment, the linker-fatty acid is εK-γE-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-CO-(CH2)18-CO2H. In a specific embodiment comprising a polypeptide or a pharmaceutically acceptable salt thereof of formula Id, K at X35 is bound to the linker-fatty acid. In another embodiment, the polypeptide comprises SEQ ID NO: 35. In another embodiment, the polypeptide is SEQ ID NO: 35. Alternatively, the polypeptide may have at least about 90% to 97% sequence similarity to SEQ ID NO: 35. Alternatively, the polypeptide may have at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, or 97% sequence similarity to SEQ ID NO: 35. An exemplary embodiment comprising SEQ ID NO: 35 is shown.

[0179] In a specific embodiment comprising a polypeptide or a pharmaceutically acceptable salt thereof of formula Id, X15 is T, X16 is G, and the polypeptide further comprises a linker-fatty acid. In a specific embodiment, the linker-fatty acid is γE-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-εK-CO-(CH2)18-CO2H. In a specific embodiment comprising a polypeptide or a pharmaceutically acceptable salt thereof of formula Id, K at X35 is bound to the linker-fatty acid. In another embodiment, the polypeptide comprises SEQ ID NO: 36. In another embodiment, the polypeptide is SEQ ID NO: 36. Alternatively, the polypeptide may have at least about 90% to 97% sequence similarity to SEQ ID NO: 36. Alternatively, the polypeptide may have at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, or 97% sequence similarity to SEQ ID NO: 36. An exemplary sequence comprising SEQ ID NO: 36 is shown.

[0180] In a specific embodiment comprising a polypeptide or a pharmaceutically acceptable salt thereof of formula Id, X15 is T, X16 is G, and the polypeptide further comprises a linker-fatty acid. In a specific embodiment, the linker-fatty acid is γE-εK-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-CO-(CH2)18-CO2H. In a specific embodiment comprising a polypeptide or a pharmaceutically acceptable salt thereof of formula Id, K at X35 is bound to the linker-fatty acid. In another embodiment, the polypeptide comprises SEQ ID NO: 37. In another embodiment, the polypeptide is SEQ ID NO: 37. Alternatively, the polypeptide may have at least about 90% to 97% sequence similarity to SEQ ID NO: 37. Alternatively, the polypeptide may have at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, or 97% sequence similarity to SEQ ID NO: 37. An exemplary sequence comprising SEQ ID NO: 37 is shown.

[0181] In a particular embodiment comprising a polypeptide or a pharmaceutically acceptable salt thereof, X15 is T, X16 is G, and the polypeptide further comprises a linker-fatty acid. In a particular embodiment, the linker-fatty acid is γE-γE-εK-CO-(CH2)18-CO2H. In a particular embodiment comprising a polypeptide or a pharmaceutically acceptable salt thereof, K at X35 is bound to the linker-fatty acid. In another embodiment, the polypeptide comprises SEQ ID NO: 38. In another embodiment, the polypeptide is SEQ ID NO: 38. Alternatively, the polypeptide may have at least about 90% to 97% sequence similarity to SEQ ID NO: 38. Alternatively, the polypeptide may have at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, or 97% sequence similarity to SEQ ID NO: 38. An exemplary embodiment comprising SEQ ID NO: 38 is shown.

[0182] In a particular embodiment comprising a polypeptide or a pharmaceutically acceptable salt thereof, X15 is T, X16 is G, and the polypeptide further comprises a linker-fatty acid. In a particular embodiment, the linker-fatty acid is γE-εK-γE-CO-(CH2)18-CO2H. In a particular embodiment comprising a polypeptide or a pharmaceutically acceptable salt thereof, K at X35 is bound to the linker-fatty acid. In another embodiment, the polypeptide comprises SEQ ID NO: 39. In another embodiment, the polypeptide is SEQ ID NO: 39. Alternatively, the polypeptide may have at least about 90% to 97% sequence similarity to SEQ ID NO: 39. Alternatively, the polypeptide may have at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, or 97% sequence similarity to SEQ ID NO: 39. An exemplary embodiment comprising SEQ ID NO: 39 is shown.

[0183] In a particular embodiment comprising a polypeptide or a pharmaceutically acceptable salt thereof, X15 is T, X16 is G, and the polypeptide further comprises a linker-fatty acid. In a particular embodiment, the linker-fatty acid is εK-γE-γE-CO-(CH2)18-CO2H. In a particular embodiment comprising a polypeptide or a pharmaceutically acceptable salt thereof, K at X35 is bound to the linker-fatty acid. In another embodiment, the polypeptide comprises SEQ ID NO: 40. In another embodiment, the polypeptide is SEQ ID NO: 40. Alternatively, the polypeptide may have at least about 90% to 97% sequence similarity to SEQ ID NO: 40. Alternatively, the polypeptide may have at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, or 97% sequence similarity to SEQ ID NO: 40. An exemplary embodiment comprising SEQ ID NO: 40 is shown.

[0184] In a specific embodiment comprising a polypeptide or a pharmaceutically acceptable salt thereof, X15 is T, X16 is Aib, and the polypeptide further comprises a linker-fatty acid. In a specific embodiment, the linker-fatty acid is 2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)2-γE-CO-(CH2)18-CO2H. In a specific embodiment comprising a polypeptide or a pharmaceutically acceptable salt thereof, K at X35 is bound to the linker-fatty acid. In another embodiment, the polypeptide comprises SEQ ID NO: 41. In another embodiment, the polypeptide is SEQ ID NO: 41. Alternatively, the polypeptide may have at least about 90% to 97% sequence similarity to SEQ ID NO: 41. Alternatively, the polypeptide may have at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, or 97% sequence similarity to SEQ ID NO: 41. An exemplary embodiment comprising SEQ ID NO: 41 is shown.

[0185] In a particular embodiment comprising a polypeptide or a pharmaceutically acceptable salt thereof of formula Id, X15 is I, X16 is Aib, and the polypeptide further comprises a linker-fatty acid moiety. In a particular embodiment, the linker-fatty acid is 2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)2-γE-CO-(CH2)18-CO2H. In a particular embodiment comprising a polypeptide or a pharmaceutically acceptable salt thereof of formula Id, K at X35 is bound to the linker-fatty acid. In another embodiment, the polypeptide comprises SEQ ID NO: 42. In another embodiment, the polypeptide is SEQ ID NO: 42. Alternatively, the polypeptide may have at least about 90% to 97% sequence similarity to SEQ ID NO: 42. Alternatively, the polypeptide may have at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, or 97% sequence similarity to SEQ ID NO: 42. An exemplary embodiment comprising SEQ ID NO: 42 is shown.

[0186] In a particular embodiment comprising a polypeptide or a pharmaceutically acceptable salt thereof, X15 is T, X16 is G, and the polypeptide further comprises a linker-fatty acid. In a particular embodiment, the linker-fatty acid is GKEKEKE-CO-(CH2)18-CO2H. In a particular embodiment comprising a polypeptide or a pharmaceutically acceptable salt thereof, K at X35 is bound to the linker-fatty acid. In another embodiment, the polypeptide comprises SEQ ID NO: 43. In another embodiment, the polypeptide is SEQ ID NO: 43. Alternatively, the polypeptide may have at least about 90% to 97% sequence similarity to SEQ ID NO: 43. Alternatively, the polypeptide may have at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, or 97% sequence similarity to SEQ ID NO: 43. An exemplary embodiment comprising SEQ ID NO: 43 is shown.

[0187] In a particular embodiment comprising a polypeptide or a pharmaceutically acceptable salt thereof, X15 is T, X16 is G, and the polypeptide further comprises a linker-fatty acid. In a particular embodiment, the linker-fatty acid is 2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-γE2-CO-(CH2)18-CO2H. In a particular embodiment comprising a polypeptide or a pharmaceutically acceptable salt thereof, K at X35 is bound to the linker-fatty acid. In another embodiment, the polypeptide comprises SEQ ID NO: 44. In another embodiment, the polypeptide is SEQ ID NO: 44. Alternatively, the polypeptide may have at least about 90% to 97% sequence similarity to SEQ ID NO: 44. Alternatively, the polypeptide may have at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, or 97% sequence similarity to SEQ ID NO: 44. An exemplary embodiment comprising SEQ ID NO: 44 is shown.

[0188] In a particular embodiment comprising a polypeptide or a pharmaceutically acceptable salt thereof, X15 is I, X16 is Aib, and the polypeptide further comprises a linker-fatty acid. In a particular embodiment, the linker-fatty acid is γE2-CO-(CH2)18-CO2H. In a particular embodiment comprising a polypeptide or a pharmaceutically acceptable salt thereof, K at X35 is bound to the linker-fatty acid. In another embodiment, the polypeptide comprises SEQ ID NO: 51. In another embodiment, the polypeptide is SEQ ID NO: 51. Alternatively, the polypeptide may have at least about 90% to 97% sequence similarity to SEQ ID NO: 51. Alternatively, the polypeptide may have at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, or 97% sequence similarity to SEQ ID NO: 51. An exemplary embodiment comprising SEQ ID NO: 51 is shown.

[0189] In another embodiment of Formula I, X1 is absent, X2 is absent, X3 is absent, X4 is P, X5 is P, X6 is P, X11 is αMeL, X13 is V or D-Val, X15 is T, X16 is Aib, X17 is αMeL, X19 is Q, X20 is K, X21 is L or Aib, X22 is L, X26 is R, X27 is A, X30 is A, X31 is R, X35 is K, X37 is A or N, and X38 is A or T. In such embodiments, the polypeptide comprises a sequence of 41 amino acids.

[0190] In some embodiments of the present invention, the polypeptide or a pharmaceutically acceptable salt thereof comprises formula Ie, which includes PPPIVTS αMeL DX13PTAib αMeL LQKX21LEQERAEKARQQAKEX37X38EILAQV (SEQ ID NO: 73), where X13 is V or D-Val [where X1 to X3 are absent, X13 is located at position 10 of the polypeptide], X21 is either L or Aib [where X21 is at position 18 of the polypeptide when X1 to X3 are absent]. X37 is either A or N [where X37 is located at position 34 of the polypeptide when X1 to X3 are absent]. X38 is either A or T [where X38 is located at position 35 of the polypeptide when X1 to X3 are absent]. The N-terminal amino acid is N-acetylated as appropriate, and the C-terminal amino acid is amylated as appropriate.

[0191] In a particular embodiment comprising a polypeptide or a pharmaceutically acceptable salt thereof of formula Ie, X13 is D-Val, X21 is Aib, X37 is N, X38 is T, and the polypeptide further comprises a linker-fatty acid. In a particular embodiment, the linker-fatty acid is γE-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-εK-CO-(CH2)18-CO2H. In a particular embodiment, K at X35 is bound to the linker-fatty acid. In some embodiments, the polypeptide comprises SEQ ID NO: 46. In some embodiments, the polypeptide is SEQ ID NO: 46. Alternatively, the polypeptide may have at least about 90% to 97% sequence similarity to SEQ ID NO: 46. Alternatively, the polypeptide may have at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, or 97% sequence similarity to SEQ ID NO: 46. An exemplary embodiment comprising SEQ ID NO: 46 is shown.

[0192] In a specific embodiment comprising a polypeptide or a pharmaceutically acceptable salt thereof of formula Id, X13 is D-Val, X21 is L, X37 is A, X38 is A, and the polypeptide further comprises a linker-fatty acid. In a specific embodiment, the linker-fatty acid is γE-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-εK-CO-(CH2)18-CO2H. In a specific embodiment comprising a polypeptide or a pharmaceutically acceptable salt thereof of formula Id, K at X35 is bound to the linker-fatty acid. In some embodiments, the polypeptide comprises SEQ ID NO: 47. In some embodiments, the polypeptide is SEQ ID NO: 47. Alternatively, the polypeptide may have at least about 90% to 97% sequence similarity to SEQ ID NO: 47. Alternatively, the polypeptide may have at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, or 97% sequence similarity to SEQ ID NO: 47. An exemplary embodiment containing SEQ ID NO: 47 is shown.

[0193] In a particular embodiment comprising a polypeptide or a pharmaceutically acceptable salt thereof, X13 is V, X21 is L, X37 is A, X38 is A, and the polypeptide further comprises a linker fatty acid. In one particular embodiment, the linker-fatty acid is (γE-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-εK-CO-(CH2)18-CO2H. In one particular embodiment comprising a polypeptide or a pharmaceutically acceptable salt thereof, K at X35 is bound to the linker-fatty acid. In some embodiments, the polypeptide comprises SEQ ID NO: 48. In some embodiments, the polypeptide is SEQ ID NO: 48. Alternatively, the polypeptide may have at least about 90% to 97% sequence similarity to SEQ ID NO: 48. Alternatively, the polypeptide may have at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, or 97% sequence similarity to SEQ ID NO: 48. An exemplary embodiment comprising SEQ ID NO: 48 is shown.

[0194] In another embodiment of Formula I, X1 is γE, X2 is absent, X3 is absent; X4 is G, X5 is P, X6 is S, X11 is αMeL, X13 is D-Val, X15 is I, X16 is Aib, X17 is αMeL, X19 is Q, X20 is I, X21 is K, X22 is L, X26 is R, X27 is A, X30 is A, X31 is R, X35 is K, X37 is N, and X38 is T. In such embodiments, the polypeptide comprises a sequence of 42 amino acids. In a particular embodiment, the polypeptide further comprises a linker-fatty acid. In a particular embodiment, the linker-fatty acid is (2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)2-γE-CO-(CH2)18-CO2H. In a particular embodiment comprising a polypeptide or a pharmaceutically acceptable salt thereof, K at X35 is bound to a linker-fatty acid.

[0195] In some embodiments, the polypeptide comprises SEQ ID NO: 45. In some embodiments, the polypeptide is SEQ ID NO: 45. Alternatively, the polypeptide may have at least about 90% to 97% sequence similarity to SEQ ID NO: 45. Alternatively, the polypeptide may have at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, or 97% sequence similarity to SEQ ID NO: 45. An exemplary embodiment comprising SEQ ID NO: 45 is shown.

[0196] In another embodiment of Formula I, X1 is absent, X2 is absent, X3 is absent, X4 is absent, X5 is absent, X6 is absent, X11 is L, X13 is V, X15 is I, X16 is G, X17 is αMeL, X19 is Q, X20 is K, X21 is L, X22 is I, X26 is K, X27 is Q, X30 is E, X31 is K, X35 is K, X37 is N, and X38 is T. In this embodiment, the polypeptide comprises a sequence of 38 amino acids.

[0197] In a particular embodiment of the polypeptide or a pharmaceutically acceptable salt thereof, the polypeptide further comprises a linker-fatty acid. In a particular embodiment, the linker-fatty acid is (2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)2γE-C20-OH)). In a particular embodiment comprising the polypeptide or a pharmaceutically acceptable salt thereof, K at X35 is bound to the linker-fatty acid.

[0198] In some embodiments, the polypeptide comprises SEQ ID NO: 49. In some embodiments, the polypeptide is SEQ ID NO: 49. Alternatively, the polypeptide may have at least about 90% to 97% sequence similarity to SEQ ID NO: 49. Alternatively, the polypeptide may have at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, or 97% sequence similarity to SEQ ID NO: 49. Exemplary embodiments comprising SEQ ID NO: 49 are shown.

[0199] In another embodiment of Formula I, X1 is absent, X2 is absent, X3 is absent, X4 is absent, X5 is absent, X6 is absent, X11 is L, X13 is V, X15 is I, X16 is G, X17 is αMeL, X19 is Q, X20 is K, X21 is L, X22 is L, X26 is R, X27 is A, X30 is A, X31 is R, X35 is K, X37 is A, and X38 is A. In this embodiment, the polypeptide comprises a sequence of 38 amino acids.

[0200] In a particular embodiment of the polypeptide or a pharmaceutically acceptable salt thereof, the polypeptide further comprises a linker-fatty acid. In one particular embodiment, the linker-fatty acid is 2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)2-γE-CO-(CH2)18-CO2H. In a particular embodiment comprising the polypeptide or a pharmaceutically acceptable salt thereof, K at X35 is bound to the linker-fatty acid.

[0201] In some embodiments, the polypeptide comprises SEQ ID NO: 50. In other embodiments, the polypeptide is SEQ ID NO: 50. Alternatively, the polypeptide may have at least about 90% to 97% sequence similarity to SEQ ID NO: 50. Alternatively, the polypeptide may have at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, or 97% sequence similarity to SEQ ID NO: 50. Exemplary embodiments comprising SEQ ID NO: 50 are shown.

[0202] In one embodiment, this document provides a polypeptide or a pharmaceutically acceptable salt thereof comprising a sequence having at least eight amino acid modifications compared to human urocortin-2 (SEQ ID NO: 53). In some embodiments, the polypeptide provided herein comprises a sequence having at least one amino acid modification at positions 3, 19, 22, 23, 26, 30, 33, or 37 of human urocortin-2 (SEQ ID NO: 53). In some embodiments, the polypeptide provided herein comprises a sequence having at least one of the following amino acid modifications compared to human urocortin-2 (SEQ ID NO: 53): L3T, A19E, R22E, A23K, E26Q, T30E, R33E, R37Q, or any combination thereof.

[0203] In some embodiments, the present invention provides a polypeptide comprising any of SEQ ID NO: 1 to 52 or a pharmaceutically acceptable salt thereof. Alternatively, the polypeptide may have at least about 90% to 99% sequence similarity to any of SEQ ID NO: 1 to 52 or a pharmaceutically acceptable salt thereof. Alternatively, the polypeptide may have at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence similarity to any of SEQ ID NO: 1 to 52.

[0204] In one specific embodiment, the polypeptide comprises SEQ ID NO: 1 or a pharmaceutically acceptable salt thereof. In one specific embodiment, the polypeptide comprises SEQ ID NO: 2 or a pharmaceutically acceptable salt thereof. In one specific embodiment, the polypeptide comprises SEQ ID NO: 22 or a pharmaceutically acceptable salt thereof. In one specific embodiment, the polypeptide comprises SEQ ID NO: 25 or a pharmaceutically acceptable salt thereof. In one specific embodiment, the polypeptide comprises SEQ ID NO: 29 or a pharmaceutically acceptable salt thereof. In one specific embodiment, the polypeptide comprises SEQ ID NO: 41 or a pharmaceutically acceptable salt thereof. In one specific embodiment, the polypeptide comprises SEQ ID NO: 42 or a pharmaceutically acceptable salt thereof. In one specific embodiment, the polypeptide comprises SEQ ID NO: 45 or a pharmaceutically acceptable salt thereof. In one specific embodiment, the polypeptide comprises SEQ ID NO: 46 or a pharmaceutically acceptable salt thereof. In one specific embodiment, the polypeptide comprises SEQ ID NO: 48 or a pharmaceutically acceptable salt thereof. In one particular embodiment, the polypeptide comprises SEQ ID NO: 49 or a pharmaceutically acceptable salt thereof. In another particular embodiment, the polypeptide comprises SEQ ID NO: 50 or a pharmaceutically acceptable salt thereof.

[0205] In a particular embodiment, the polypeptide or its pharmaceutically acceptable salt system is selected from the group consisting of: SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 22, SEQ ID NO: 25, SEQ ID NO: 29, SEQ ID NO: 41, SEQ ID NO: 42, SEQ ID NO: 45, SEQ ID NO: 46, SEQ ID NO: 48, SEQ ID NO: 49, and SEQ ID NO: 50.

[0206] In certain embodiments of the present invention, the pharmaceutically acceptable salts are selected from sodium, potassium, trifluoroacetate, hydrochloride, or acetate.

[0207] In some embodiments, the present invention provides a pharmaceutical composition comprising a polypeptide or a pharmaceutically acceptable salt thereof and at least one pharmaceutically acceptable carrier, diluent or excipient.

[0208] In some embodiments, the present invention provides a pharmaceutical composition wherein the composition is formulated for oral administration. In other embodiments, the composition is formulated for subcutaneous administration.

[0209] In some embodiments, the present invention provides a method for treating diseases or conditions selected from the group consisting of: diabetes, obesity, chronic weight management, non-alcoholic fatty liver disease (NAFLD), non-alcoholic steatosis (NASH), dyslipidemia, metabolic syndrome, chronic kidney disease (CKD), osteoarthritis (OA), obesity-related sleep apnea (OSA), sarcopenia, cachexia, sarcopenic obesity (SO), and polycystic ovary syndrome (PCOS), the method comprising administering an effective amount of a polypeptide or a medically acceptable salt thereof to an individual in need.

[0210] In some embodiments, the present invention provides a polypeptide or a pharmaceutically acceptable salt thereof for use in a therapy.

[0211] In some embodiments, the present invention provides a polypeptide or a pharmaceutically acceptable salt thereof for the treatment of diseases or conditions selected from the group consisting of: diabetes, obesity, chronic weight management, non-alcoholic fatty liver disease (NAFLD), non-alcoholic steatohepatitis (NASH), dyslipidemia, metabolic syndrome, chronic kidney disease (CKD), osteoarthritis (OA), obesity-related sleep apnea (OSA), sarcopenia, cachexia, sarcopenic obesity (SO), and polycystic ovary syndrome (PCOS).

[0212] In some embodiments, the present invention provides the use of polypeptides or pharmaceutically acceptable salts thereof for the manufacture of medicaments for treating diseases or conditions selected from the group consisting of: diabetes, obesity, chronic weight management, non-alcoholic fatty liver disease (NAFLD), non-alcoholic steatohepatitis (NASH), dyslipidemia, metabolic syndrome, chronic kidney disease (CKD), osteoarthritis (OA), obesity-related sleep apnea (OSA), sarcopenia, cachexia, sarcopenic obesity (SO), and polycystic ovary syndrome (PCOS).

[0213] In some embodiments, the present invention provides a method for treating diseases or conditions selected from the group consisting of: diabetes, obesity, chronic weight management, non-alcoholic fatty liver disease (NAFLD), non-alcoholic steatosis (NASH), dyslipidemia, metabolic syndrome, chronic kidney disease (CKD), osteoarthritis (OA), obesity-related sleep apnea (OSA), sarcopenia, cachexia, sarcopenic obesity (SO), and polycystic ovary syndrome (PCOS), the method comprising administering to the patient an effective amount of a polypeptide and an effective amount of an additional therapeutic agent.

[0214] In one embodiment, this document provides a polypeptide or a pharmaceutically acceptable salt thereof, which can be used simultaneously, separately, or sequentially in combination with one or more additional therapeutic agents.

[0215] In some embodiments, the present invention provides a method for increasing athletic ability or maximum VO2.

[0216] In some embodiments, the additional therapeutic agent is a dual agonist of the GIP and GLP-1 receptors. In a particular embodiment, the dual agonist of the GIP and GLP-1 receptors is a compound comprising SEQ ID NO: 55.

[0217] In some embodiments, the additional therapeutic agent is a triple agonist of GIP, GLP-1, and glucagon receptor. In a particular embodiment, the triple agonist of GIP, GLP-1, and glucagon receptor is a compound comprising SEQ ID NO: 56, SEQ ID NO: 57, or SEQ ID NO: 58, or a pharmaceutically acceptable salt thereof.

[0218] In some embodiments, the present invention provides a method wherein the additional therapeutic agent is a dual agonist of glucagon and GLP-1 receptor. In a particular embodiment, the dual agonist of glucagon and GLP-1 receptor is a compound comprising SEQ ID NO: 59, SEQ ID NO: 60, SEQ ID NO: 61 or SEQ ID NO: 62 or a pharmaceutically acceptable salt thereof.

[0219] In some embodiments, the additional therapeutic agent is a agonist selective for amyloid receptors. In a particular embodiment, the agonist selective for amyloid receptors is a compound comprising SEQ ID NO: 63, SEQ ID NO: 64, SEQ ID NO: 65, or SEQ ID NO: 66.

[0220] In some embodiments, the additional therapeutic agent is a dual agonist of calcitonin and amyloid receptors. In a particular embodiment, the dual agonist of calcitonin and amyloid receptors is a compound comprising SEQ ID NO: 67.

[0221] In some embodiments, the additional therapeutic agent is a GLP-1 receptor agonist. In a particular embodiment, the GLP-1 receptor agonist is a compound comprising SEQ ID NO: 79, SEQ ID NO: 80, or SEQ ID NO: 81.

[0222] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one skilled in the art to which this invention pertains. Although any methods and materials similar to or equivalent to those described herein may be used to practice or test peptides, pharmaceutical compositions, and methods, preferred methods and materials are described herein.

[0223] Furthermore, unless the context explicitly requires the existence of one and only one element, mentioning an element by the indefinite article "a" or "an" does not preclude the possibility of more than one element. Therefore, the indefinite article "a" or "an" generally means "at least one".

[0224] As used herein, “about” means a statistically significant range of one or more values ​​(such as specified concentration, length, molecular weight, pH, sequence consistency, time range, temperature, or volume). Such values ​​or ranges may be within one order of magnitude of a given value or range, typically within 20%, more typically within 10%, and even more typically within 5%. The permissible variation covered by “about” will depend on the specific system studied and will be readily understood by one skilled in the art.

[0225] As used herein, "additional therapeutic agent" refers to a substance used to treat, manage, or prevent disease that is different from a polypeptide active against the CRH2 receptor. In this context, additional therapeutic agents may include (but are not limited to) dual agonists of GIP and GLP-1 receptors, triple agonists of GIP, GLP-1, and glucagon receptors, dual agonists of glucagon and GLP-1 receptors, agonists selective for amyloid receptors, dual agonists of calcitonin and amyloid receptors, agonists of the GLP-1 receptor, or leptin receptor antibodies.

[0226] As used herein, and for the purposes of CRH (e.g., CRHR1 or CRHR2) receptors, one or more of the terms "activity," "activate," "activating," and similar expressions mean the ability of a compound (such as the peptides described herein) to bind to the receptor and induce a response at the receptor, measured using analytical methods known in this art, such as in vitro analyses described below.

[0227] As used herein, "an amino acid having a functional group available for binding" means any natural (encoded) or non-natural (non-encoded) amino acid having a functional group that can be bound to a fatty acid directly or by means of, for example, a linker. Examples of such functional groups include (but are not limited to) alkynyl, alkenyl, amino, azide, bromo, carboxyl, chloro, iodo, and mercapto. Examples of natural amino acids including such functional groups include K (amino), C (mercapto), E (carboxyl), and D (carboxyl).

[0228] As used herein, "conservative amino acid substitution" means an amino acid that has similar characteristics (e.g., charge, side chain size, hydrophobicity / hydrophilicity, main chain configuration, and rigidity) and has minimal impact on the bioactivity of the resulting substituted peptide or polypeptide. Conservative substitution of functionally similar amino acids is well known in this art and therefore need not be described in detail herein. As used in this article, "at least" means no less than; minimum.

[0229] As used in this article, Body Mass Index (BMI) refers to an individual's weight in kilograms divided by the square of their height in meters.

[0230] As used in this article, "cachexia" refers to a complex syndrome associated with an underlying disease that leads to persistent muscle loss that cannot be fully reversed by nutritional supplementation. A range of diseases can trigger cachexia, including (but not limited to) cancer, congestive heart failure, chronic obstructive pulmonary disease, chronic kidney disease, and AIDS.

[0231] As used herein, "chronic weight management" refers to the long-term management of the weight of overweight or obese individuals. Chronic weight management can be achieved through the peptides of this invention. In addition to healthy lifestyle changes (including, but not limited to, a balanced diet and regular physical activity), chronic weight management can also be achieved through the peptides of this invention.

[0232] As used in this article, "chronic kidney disease" (CKD) refers to the loss of kidney function over time. Due to the loss of kidney function, individuals with CKD may be affected by one or more of the following symptoms or signs: nausea, vomiting, loss of appetite, fatigue, weakness, sleep problems, excessive urination, muscle cramps, swelling of the feet or ankles, high blood pressure, or shortness of breath.

[0233] As used herein, "conservative amino acid substitution" means an amino acid that has similar characteristics (e.g., charge, side chain size, hydrophobicity / hydrophilicity, main chain configuration, and rigidity) and has minimal impact on the bioactivity of the resulting substituted peptide or polypeptide. Conservative substitution of functionally similar amino acids is well known in this art and therefore need not be described in detail herein.

[0234] As used herein, "C16-C22 fatty acid" refers to a carboxylic acid having between 16 and 22 carbon atoms. The C16-C22 fatty acid used herein can be a saturated monocarboxylic acid or a saturated diacid. As used herein, "saturated" means a fatty acid that does not contain carbon-carbon double bonds or triple bonds.

[0235] As used herein, "diabetes" or "diabetes mellitus" refers to a group of endocrine disorders characterized by high blood sugar levels. In this context, diabetes refers to a condition where the pancreas fails to produce sufficient insulin, or where the body's cells become unresponsive to hormones. Diabetes also refers to type 2 diabetes (T2DM), characterized by high blood sugar, insulin resistance, and / or relative insulin deficiency. As used herein, diabetes also refers to high blood sugar levels in women during pregnancy, known as gestational diabetes.

[0236] As used in this article, "dyslipidemia" refers to an abnormally high level of lipids (fat) in the blood. Dyslipidemia also refers to a high level of cholesterol in the blood.

[0237] As used herein, "effective dose" means the amount, concentration, or dosage of one or more polypeptides or their pharmaceutically acceptable salts described herein that, after administration of a single or multiple doses to an individual in need, produce the desired effect on that individual undergoing diagnosis or treatment. Effective doses can be readily determined by a person skilled in the art using known techniques and by observations obtained under similar conditions. In determining the effective dose for an individual, various factors are considered, including (but not limited to) the mammalian species; its size, age, and general health status; the specific disease or condition involved; the extent or severity of the disease or condition; the individual patient's response; the specific polypeptide administered; the administration pattern; the bioavailability characteristics of the administered formulation; the chosen dosing regimen; the use of concomitant medications; and other relevant factors.

[0238] As used herein, "extended duration of action" means that the binding affinity and activity of the peptide last for a longer period than that of natural human urocorticoids, thereby allowing for dosing frequencies as low as at least once daily or even three, two, or once weekly. The duration of action of the peptide can be measured using known pharmacokinetic assays, such as those used in the examples below.

[0239] As used in this article, “metabolic syndrome” refers to a group of health conditions that increase the risk of heart disease and related problems. Individuals with metabolic syndrome have three or more of the following health conditions: (1) high blood sugar, (2) low HDL cholesterol, (3) high triglyceride levels in the blood, (4) large waist circumference, and (5) high blood pressure.

[0240] As used herein, "NASH" refers to non-alcoholic fatty liver disease, also known as fatty liver disease. "NASH" also refers to liver inflammation and damage caused by fat accumulation in the liver. "NASH" is also a subtype of non-alcoholic fatty liver disease ("NAFLD"). In some embodiments, "NASH" may be synonymous with "NAFLD".

[0241] As used in this article, “obesity” refers to a condition involving an increased risk of health problems due to excess body fat. The term “obesity” also refers to a weight exceeding what is considered a healthy weight for a given height. The term “obesity” also refers to a BMI greater than 30.0, or a BMI of 27.0 or greater (overweight), accompanied by at least one weight-related comorbidity (such as hypertension, type 2 diabetes, or dyslipidemia).

[0242] As used in this article, "osteoarthritis" (OA) is a form of arthritis that causes inflammation of one or more joints. "Osteoarthritis" can refer to arthritis affecting the joints of the hands, spine, knees, and hips.

[0243] As used herein, "sleep apnea" (OSA) refers to a sleep-related breathing disorder characterized by recurrent episodes of complete or partial obstruction of the upper airway, resulting in reduced or absent breathing during sleep.

[0244] As used herein, "patient" and "individual" are used interchangeably and refer to mammals, and preferably humans. In some embodiments, a patient, preferably a human, is further characterized as suffering from a disease, condition, or symptom that would benefit from administration of the polypeptide of the present invention.

[0245] As used herein, "polypeptide" or "peptide" means a polymer of amino acid residues. This term applies to polymers containing naturally occurring amino acids as well as polymers containing one or more non-naturally occurring amino acids.

[0246] As used in this article, "polycystic ovary syndrome" (PCOS) refers to an endocrine disorder in women of reproductive age. PCOS also refers to the fact that a woman may have one or more of the following symptoms: ovarian cysts, irregular menstrual periods, heavy menstrual bleeding, excessive hair growth, difficulty conceiving, or insulin resistance.

[0247] As used in this article, "individual in need" means a mammal, such as a human, that suffers from a condition, disease, ailment, or symptom requiring treatment or therapy (including, for example, those listed in this article).

[0248] As used in this article, "quality weight loss" refers to a reduction in fat mass while maintaining lean body mass.

[0249] As used in this article, "sarcopenia" refers to a progressive and systemic skeletal muscle disorder involving an accelerated loss of muscle mass and function.

[0250] As used in this article, "sarcopenic obesity" refers to an individual who suffers from both sarcopenia (muscle loss) and obesity. Sarcopenic obesity also refers to the presence of low muscle mass or low muscle strength and high fat mass.

[0251] As used herein, the term "sequence similarity" refers to the degree of similarity between two sequences. The degree of sequence similarity between two polypeptides can be expressed as a percentage, calculated as follows:

[0252] Sequence identity % = 100% (number of identical amino acids) / (length of the shortest common sequence)

[0253] As used in this article, "treat / treating / to treat" and similar expressions mean to limit, slow down, stop or reverse the progression or severity of an existing symptom, disease, condition or illness. Additional non-limiting embodiments are described below. 1. A polypeptide or a pharmaceutically acceptable salt thereof, comprising: X1X2X3X4X5X6IVTSX11DX13PX15X16X17LX19X20X21X22EQEX26X27EKX30X31QQAX35EX37X38EILAQV (SEQ ID NO: 68), where X1 is Pyr, γE, or does not exist. X2 is G or does not exist. X3 is G, S, or does not exist. X4 is S, P, G, or does not exist. X5 is S, P, or does not exist. X6 is G, S, P, or does not exist. X11 is L or αMeL, X13 is either V or D-Val. X15 is either T or I. X16 is either Aib or G. X17 is αMeL. X19 is either Q or E. X20 is either K or I. X21 can be I, K, Aib, or L. X22 represents I or L. X26 is either R or K. X27 is either A or Q. X30 is either A or E. X31 is either R or K. X35 is either K or T. X37 is either A or N, and X38 is either A or T. 2. The polypeptide of Example 1 or a pharmaceutically acceptable salt thereof, comprising: GSPSIVTS αMeL DVPT Aib αMeL LQKX21LEQERAEKARQQAKEAAEILAQV (SEQ ID NO: 69), where X21 is either I or L. The N-terminal amino acid is G at X3, and G is N-acetylated, and the C-terminal amino acid is V, and V is amylated. 3. The polypeptide of Example 2 or a pharmaceutically acceptable salt thereof, further comprising a linker-fatty acid bound to the ε-amino group of the side chain of K at X35, wherein the binding system of K to the linker-fatty acid is selected from the group consisting of: K((2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-PKE-γE-CO-(CH2)18-CO2H), K(Ahx-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-γE-CO-(CH2)18-CO2H), K(γE-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-γE-CO-(CH2)18-CO2H), K(γE-(2-[2-(2-amino-ethoxy)-acetyl ...acetyl)-acetyl)-acetyl)-acetyl)-acetyl)-acetyl)-acetyl)-acetyl)-acetyl)-acetyl)-acetyl)-acetyl)-acetyl)-acetyl)-acetyl)-acetyl)-acetyl)-acetyl)-acetyl) [2-(2-amino-ethoxy)-ethoxy]-acetyl)-Ahx-CO-(CH2)18-CO2H), K(γE-GKEKEKE-CO-(CH2)18-CO2H), K(G-(βA)3-γE-CO-(CH2)18-CO2H), K(G-(βA)4-γE-CO-(CH2)18-CO2H), K((2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-KEKEKE-γE-CO-(CH2)18-CO2H), K(βA-S-βA-S-γE-CO-(C H2)18-CO2H), K(γE-PKE-Ahx-CO-(CH2)18-CO2H), K(γE-PKE-εK-CO-(CH2)18-CO2H), K(γE-PPP-εK-CO-(CH2)18-CO2H), K(γE-PPPPPP-CO-(CH2)18-CO2H), K(γE-PEPEPE-CO-(CH2)18-CO2H), K(γE-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)2-εK-CO-(CH2) K(γE-εK-γE-εK-CO-(CH2)18-CO2H), K(γE-εK-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-εK-CO-(CH2)18-CO2H), K(γE-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-εK-CO-(CH2)18-CO2H), and K(γE-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-εK-CO-(CH2)16-CO2H). 4. The polypeptide of Example 3 or a pharmaceutically acceptable salt thereof, wherein X21 is I, and K at X35 is bound to a fatty acid according to the formula K(γE-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-εK-CO-(CH2)18-CO2H). 5. The polypeptide of Example 3 or a pharmaceutically acceptable salt thereof, wherein X21 is L, and K at X35 is bound to a fatty acid according to the formula K(γE-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-εK-CO-(CH2)18-CO2H). 6. The polypeptide of Example 3 or a pharmaceutically acceptable salt thereof, wherein X21 is I, and K at X35 is bound to a fatty acid according to the formula K((2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-PKE-γE-CO-(CH2)18-CO2H). 7. The polypeptide of Example 3 or a pharmaceutically acceptable salt thereof, wherein X21 is I, and K at X35 is bound to a fatty acid according to the formula K(Ahx-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-γE-CO-(CH2)18-CO2H). 8. The polypeptide of Example 3 or a pharmaceutically acceptable salt thereof, wherein X21 is I, and K at X35 is bound to a fatty acid according to the formula K(γE-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-Ahx-CO-(CH2)18-CO2H). 9. The polypeptide of Example 3 or a pharmaceutically acceptable salt thereof, wherein X21 is I, and K at X35 is bound to a fatty acid according to formula K(γE-GKEKEKE-CO-(CH2)18-CO2H). 10. The polypeptide of Example 3 or a pharmaceutically acceptable salt thereof, wherein X21 is I and K at X35 is bound to a fatty acid according to formula K(G-(βA)3-γE-CO-(CH2)18-CO2H). 11. The polypeptide of Example 3 or a pharmaceutically acceptable salt thereof, wherein X21 is I and K at X35 is bound to a fatty acid according to formula K(G-(βA)4-γE-CO-(CH2)18-CO2H). 12. The polypeptide of Example 3 or a pharmaceutically acceptable salt thereof, wherein X21 is I, and K at X35 is bound to a fatty acid according to the formula K((2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-KEKEKE-γE-CO-(CH2)18-CO2H). 13. The polypeptide of Example 3 or a pharmaceutically acceptable salt thereof, wherein X21 is I and K at X35 is bound to a fatty acid according to formula K(βA-S-βA-S-γE-CO-(CH2)18-CO2H). 14. The polypeptide of Example 3 or a pharmaceutically acceptable salt thereof, wherein X21 is I and K at X35 is bound to a fatty acid according to formula K(γE-PKE-Ahx-CO-(CH2)18-CO2H). 15. The polypeptide of Example 3 or a pharmaceutically acceptable salt thereof, wherein X21 is L and K at X35 is bound to a fatty acid according to the formula K(γE-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-εK-CO-(CH2)16-CO2H). 16. The polypeptide of Example 3 or a pharmaceutically acceptable salt thereof, wherein X21 is I and K at X35 is bound to a fatty acid according to formula K(γE-PKE-εK-CO-(CH2)18-CO2H). 17. The polypeptide of Example 3 or a pharmaceutically acceptable salt thereof, wherein X21 is I and K at X35 is bound to a fatty acid according to formula K(γE-PPP-εK-CO-(CH2)18-CO2H). 18. The polypeptide of Example 3 or a pharmaceutically acceptable salt thereof, wherein X21 is I and K at X35 is bound to a fatty acid according to formula K(γE-PPPPPP-CO-(CH2)18-CO2H). 19. The polypeptide of Example 3 or a pharmaceutically acceptable salt thereof, wherein X21 is I and K at X35 is bound to a fatty acid according to formula K(γE-PEPEPE-CO-(CH2)18-CO2H). 20. The polypeptide of Example 3 or a pharmaceutically acceptable salt thereof, wherein X21 is I, and K at X35 is bound to a fatty acid according to the formula K(γE-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)2-εK-CO-(CH2)18-CO2H). 21. The polypeptide of Example 3 or a pharmaceutically acceptable salt thereof, wherein X21 is I, and K at X35 is bound to a fatty acid according to formula K(γE-εK-γE-εK-CO-(CH2)18-CO2H). 22. The polypeptide of Example 3 or a pharmaceutically acceptable salt thereof, wherein X21 is I, and K at X35 is bound to a fatty acid according to the formula K(γE-εK-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-εK-CO-(CH2)18-CO2H). 23. The polypeptide of Example 2 or a pharmaceutically acceptable salt thereof, wherein X21 is I. 24. The polypeptide of Example 3 or a pharmaceutically acceptable salt thereof, wherein X21 is I, and K at X35 is bound to a fatty acid according to the formula K(γE-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-εK-CO-(CH2)16-CO2H). 25. The polypeptide of Example 1 or a pharmaceutically acceptable salt thereof, comprising: Pyr GSPSGIVTS αMeL DVPT Aib αMeL LQKILEQERAEKARQQAKEAAEILAQV (SEQ ID NO: 70), or a pharmaceutically acceptable salt thereof, wherein the C-terminal V is acetylated. 26. The polypeptide of Example 25 or a pharmaceutically acceptable salt thereof, further comprising a linker-fatty acid bound to the ε-amino group of the side chain of K at X35, wherein the binding system of K to the linker-fatty acid is selected from the group consisting of: K(γE-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-εK-CO-(CH2)18-CO2H) and K(γE-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-εK-CO-(CH2)16-CO2H). 27. The polypeptide of Example 26, wherein K at X35 is bound to a fatty acid according to the formula K(γE-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-εK-CO-(CH2)18-CO2H). 28. The polypeptide of Example 26, wherein K at X35 is bound to a fatty acid according to the formula K(γE-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-εK-CO-(CH2)16-CO2H). 29. The polypeptide of Example 1 or a pharmaceutically acceptable salt thereof, comprising: X1GSPSIVTS αMeL DVPT Aib αMeL LX19KILEQERAEKARQQATEAAEILAQV (SEQ ID NO: 71), where X1 is either γE or does not exist. X2 does not exist. X19 is either Q or E. Furthermore, the C-terminal amino acid is V at position X44, and the V at X44 is acetylated. 30. The polypeptide of Example 29, wherein X1 is γE, and the γE at X1 is bound to G at X3 via an amide bond between the γ-carboxylic acid group of γE and the amino group of G. 31. The polypeptide of Example 30 further comprises a fatty acid linked to the α-amino group of γE and the carboxylic acid group of the (2-[2-(2-amino-ethoxy)-ethoxy]acetic acid monomer via an amide bond to γE at position X1, wherein the linker-fatty acid is selected from the group consisting of: (2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-εK-CO-(CH2)18-CO2H, (2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-K-CO-(CH2)18-CO2H, (2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-Dap-CO-(CH2)18-CO2H, and (2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-εK-CO-(CH2)16-CO2H. 32. The polypeptide of Example 31, wherein the linker-fatty acid bound to γE at X1 is (2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-εK-CO-(CH2)18-CO2H, and X19 is Q. 33. The polypeptide of Example 31, wherein the linker fatty acid that binds to γE at X1 is (2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-K-CO-(CH2)18-CO2H, and X19 is Q. 34. The polypeptide of Example 31, wherein X1 is absent and X19 is Q. 35. The polypeptide of Example 31, wherein the linker fatty acid that binds to γE at X1 is (2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-εK-CO-(CH2)16-CO2H, and X19 is Q. 36. The polypeptide of Example 31, wherein the linker fatty acid that binds to γE at X1 is (2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-Dap-CO-(CH2)18-CO2H, and X19 is E. 37. The polypeptide of Example 1 or a pharmaceutically acceptable salt thereof, comprising: IVTS αMeL D D-Val PX15X16αMeL LQIKLEQERAEKARQQAKENTEILAQV (SEQ ID NO: 72), or a medically acceptable salt thereof, wherein X15 is either T or I. X16 is either G or Aib, and The N-terminal amino acid at position X7 is I, and the I at position X7 is N-acetylated, and the C-terminal amino acid at position X44 is V, and the V at position X44 is amylated. 38. The polypeptide of Example 37, further comprising a linker-fatty acid bound to the ε-amino group of the side chain of K at X35, wherein the binding system of K to the linker-fatty acid is selected from the group consisting of: K((2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)2-γE-CO-(CH2)18-CO2H), K(γE-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-γE-CO-(CH2)18-CO2H), K(γE2-CO-(CH2)18-CO2H), K(γE2-CO-(CH2)18-CO2H). 8-CO2H), K((2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-εK-γE-CO-(CH2)18-CO2H), K((2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-γE-εK-CO-(CH2)18-CO2H), K(γE2-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-CO-(CH2)18-CO2H), K(εK-γE ...εK-γE-(2-[2-(2-amino-ethoxy)-acetyl)-εK-γE-(2-[2-(2-amino-ethoxy)-acetyl)-εK-γE-(2-[2-(2-amino-ethoxy)-acetyl)-εK-γE-(2-[2-(2-amino-ethoxy)-acetyl)-εK-γE-(2-[2-(2-amino-ethoxy)-εK-γE-(2-[2-(2-amino-ethoxy)-εK-γE-(2-[2-( K(εK-γE-γE-CO-(CH2)18-CO2H), K(εK-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-γE-CO-(CH2)18-CO2H), K(γE-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-εK-CO-(CH2)18-CO2H), K(γE-εK-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-εK-CO-(CH2)18-CO2H), K(γE-εK-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-εK-CO-(CH2)18-CO2H), K(γE-εK-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-εK-CO-(CH2)18-CO2H) K(γE-γE-εK-CO-(CH2)18-CO2H), K(γE-εK-γE-CO-(CH2)18-CO2H), K(GKEKEKE-CO-(CH2)18-CO2H), K(γE2-CO-(CH2)18-CO2H), and K((2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-γE2-CO-(CH2)18-CO2H). 39. The polypeptide of Example 38, wherein X15 is T, X16 is G, and K at X35 is bound to fatty acid according to the formula K((2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)2-γE-CO-(CH2)18-CO2H. 40. The polypeptide of Example 38, wherein X15 is T, X16 is G, and K at X35 is bound to fatty acid according to formula K(γE-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-γE-CO-(CH2)18-CO2H). 41. The polypeptide of Example 38, wherein X15 is T, X16 is G, and K at X35 is bound to fatty acid according to formula K(γE2-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-CO-(CH2)18-CO2H). 42. The polypeptide of Example 38, wherein X15 is T, X16 is G, and K at X35 is bound to fatty acid according to formula K((2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-εK-γE-CO-(CH2)18-CO2H). 43. The polypeptide of Example 38, wherein X15 is T, X16 is G, and K at X35 is bound to fatty acid according to formula K((2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-γE-εK-CO-(CH2)18-CO2H). 44. The polypeptide of Example 38, wherein X15 is T, X16 is G, and K at X35 is bound to fatty acid according to formula K(εK-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-γE-CO-(CH2)18-CO2H). 45. The polypeptide of Example 38, wherein X15 is T, X16 is G, and K at X35 is bound to fatty acid according to formula K(εK-γE-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-CO-(CH2)18-CO2H). 46. ​​The polypeptide of Example 38, wherein X15 is T, X16 is G, and K at X35 is bound to fatty acid according to formula K(γE-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-εK-CO-(CH2)18-CO2H). 47. The polypeptide of Example 38, wherein X15 is T, X16 is G, and K at X35 is bound to fatty acid according to the formula K(γE-εK-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-CO-(CH2)18-CO2H. 48. The polypeptide of Example 38, wherein X15 is T, X16 is G, and K at X35 is bound to a fatty acid according to formula K(γE-γE-εK-CO-(CH2)18-CO2H). 49. The polypeptide of Example 38, wherein X15 is T, X16 is G, and K at X35 is bound to fatty acid according to formula K(γE-εK-γE-CO-(CH2)18-CO2H). 50. The polypeptide of Example 38, wherein X15 is T, X16 is G, and K at X35 is bound to a fatty acid according to formula K(εK-γE-γE-CO-(CH2)18-CO2H). 51. The polypeptide of Example 38, wherein X15 is T, X16 is Aib, and K at X35 is bound to a fatty acid according to the formula K((2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)2-γE-CO-(CH2)18-CO2H). 52. The polypeptide of Example 38, wherein X15 is I, X16 is Aib, and K at X35 is bound to a fatty acid according to the formula K((2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)2-γE-CO-(CH2)18-CO2H). 53. The polypeptide of Example 38, wherein X15 is T, X16 is G, and K at X35 is bound to a fatty acid according to the formula K(GKEKEKE-CO-(CH2)18-CO2H). 54. The polypeptide of Example 38, wherein X15 is T, X16 is G, and K at X35 is bound to fatty acid according to formula K((2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-γE2-CO-(CH2)18-CO2H). 55. The polypeptide of Example 38, wherein X15 is I, X16 is Aib, and K at X35 is bound to a fatty acid according to formula K(γE2-CO-(CH2)18-CO2H). 56. The polypeptide of Example 1 or a pharmaceutically acceptable salt thereof, comprising: PPPIVTS αMeL DX13PTAib αMeL LQKX21LEQERAEKARQQAKEX37X38EILAQV (SEQ ID NO: 73), where, X13 is either V or D-Val. X21 is either L or Aib. X37 is either A or N. X38 is either A or T. The N-terminal amino acid at position X4 is P, and the P at position X4 is N-acetylated, and the C-terminal amino acid at position X44 is V, and the V at position X44 is acetylated. 57. The polypeptide of Example 1 or a pharmaceutically acceptable salt thereof, wherein, X1 is γE, X2 does not exist. X3 does not exist. X4 is G. X5 is P, X6 is S, X11 is αMeL. X13 is D-Val. X15 is I, X16 is Aib. X19 is Q, X20 is I, X21 is K, X22 is L, X26 is R, X27 is A. X30 is A. X31 is R, X35 is K, X37 is N, X38 is T, Furthermore, the C-terminal amino acid is V at position X44, and the V at X44 is acetylated. 58. The polypeptide of Example 56 or a pharmaceutically acceptable salt thereof, further comprising a linker-fatty acid bound to the ε-amino group of the side chain of K at X35, wherein the binding of K to the linker-fatty acid is selected from the group consisting of: K((2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)2-γE-CO-(CH2)18-CO2H) and K(γE-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-εK-CO-(CH2)18-CO2H). 59. The polypeptide of Example 58, wherein X13 is D-Val, X21 is Aib, K at X35 is bound to a fatty acid according to the formula K(γE-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-εK-CO-(CH2)18-CO2H), X37 is N, and X38 is T. 60. In the polypeptide of Example 58, X13 is D-Val, X21 is L, K at X35 is bound to fatty acid according to formula K(γE-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-εK-CO-(CH2)18-CO2H), X37 is A, and X38 is A. 61. As in Example 58, the polypeptide is V at X13, L at X21, K at X35 is bound to fatty acid according to formula K(γE-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-εK-CO-(CH2)18-CO2H), X37 is A, and X38 is A. 62. The polypeptide of Example 1 or a pharmaceutically acceptable salt thereof, wherein X1 does not exist. X2 does not exist. X3 does not exist. X4 does not exist. X5 does not exist. X6 does not exist. X11 is L, X13 is V, X15 is I, X16 is G, X19 is Q, X20 is K, X21 is L, X22 is L, X26 is K, X27 is Q, X30 is E, X31K, X35 is K, X37 is N, X38 is T, The N-terminal amino acid is I at X7, and the I at X7 is N-methylated. Furthermore, the C-terminal amino acid is V at position X44, and the V at X44 is acetylated. 63. The polypeptide of Example 62 or a pharmaceutically acceptable salt thereof, further comprising a linker-fatty acid bound to the ε-amino group of the side chain of K at X35, wherein the binding of K to the linker-fatty acid has the formula K((2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)2-γE-C20-OH). 64. The polypeptide of Example 1 or a pharmaceutically acceptable salt thereof, wherein X1 does not exist. X2 does not exist. X3 does not exist. X4 does not exist. X5 does not exist. X6 does not exist. X11 is L, X13 is V, X15 is I, X16 is G, X19 is Q, X20 is K, X21 is L, X22 is L, X26 is R, X27 is A. X30 is A. X31 is R, X35 is K, X37 is A. X38 is A. The N-terminal amino acid is I at X7, and the I at X7 is N-methylated. Furthermore, the C-terminal amino acid is V at position X44, and the V at X44 is acetylated. 65. The polypeptide of Example 64 or a pharmaceutically acceptable salt thereof, further comprising a linker-fatty acid bound to the ε-amino group of the side chain of K at X35, wherein the binding of K to the linker-fatty acid has K((2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)2-γE-CO-(CH2)18-CO2H). 66. A polypeptide comprising any one of SEQ ID NO: 1 to 52, or a pharmaceutically acceptable salt thereof. 67. The polypeptide or a pharmaceutically acceptable salt thereof as described in any of Examples 1 to 66, wherein the pharmaceutically acceptable salt is selected from sodium, potassium, trifluoroacetate, hydrochloric acid or acetate. 68. A pharmaceutical composition comprising a polypeptide or a pharmaceutically acceptable salt thereof as described in any one of Examples 1 to 66 and at least one pharmaceutically acceptable carrier, diluent or excipient. 69. The pharmaceutical composition of Example 68, wherein the composition is formulated for oral administration. 70. The pharmaceutical composition of Example 68, wherein the composition is formulated for subcutaneous administration. 71. A method for treating diseases or conditions selected from the group consisting of: diabetes, obesity, chronic weight management, non-alcoholic fatty liver disease (NAFLD), non-alcoholic steatosis (NASH), dyslipidemia, metabolic syndrome, chronic kidney disease (CKD), osteoarthritis (OA), obesity-related sleep apnea (OSA), sarcopenia, cachexia, sarcopenic obesity (SO), and polycystic ovary syndrome (PCOS), the method comprising administering to an individual in need an effective amount of a polypeptide or a pharmaceutically acceptable salt thereof, as described in any one of Examples 1 to 66. 72. The polypeptide or its pharmaceutically acceptable salt as claimed in any of Examples 1 to 66, used in a therapeutic manner. 73. The polypeptide or its pharmaceutically acceptable salt claimed in any of Examples 1 to 66, for the treatment of diseases or conditions selected from the group consisting of: diabetes, obesity, chronic weight management, non-alcoholic fatty liver disease (NAFLD), non-alcoholic steatohepatitis (NASH), dyslipidemia, metabolic syndrome, chronic kidney disease (CKD), osteoarthritis (OA), obesity-related sleep apnea (OSA), sarcopenia, cachexia, sarcopenic obesity (SO), and polycystic ovary syndrome (PCOS). 74. The use of the polypeptide or its pharmaceutically acceptable salt as claimed in any of Examples 1 to 66, for the manufacture of an agent for the treatment of diseases or conditions selected from the group consisting of: diabetes, obesity, chronic weight management, non-alcoholic fatty liver disease (NAFLD), non-alcoholic steatohepatitis (NASH), dyslipidemia, metabolic syndrome, chronic kidney disease (CKD), osteoarthritis (OA), obesity-related sleep apnea (OSA), sarcopenia, cachexia, sarcopenic obesity (SO), and polycystic ovary syndrome (PCOS). 75. A method for treating a disease or condition in a patient in need, the disease or condition being selected from the group consisting of: diabetes, obesity, chronic weight management, non-alcoholic fatty liver disease (NAFLD), non-alcoholic steatosis (NASH), dyslipidemia, metabolic syndrome, chronic kidney disease (CKD), osteoarthritis (OA), obesity-related sleep apnea (OSA), sarcopenia, cachexia, sarcopenic obesity (SO), and polycystic ovary syndrome (PCOS), the method comprising administering to the patient an effective amount of the polypeptide as described in any one of Examples 1 to 66 and an effective amount of an additional therapeutic agent. 76. The treatment method as described in Example 75, wherein the additional therapeutic agent is a dual agonist of the GIP and GLP-1 receptors. 77. The method of Example 76, wherein the dual agonist for the GIP and GLP-1 receptors is a compound comprising SEQ ID NO: 55. 78. The treatment method as described in Example 75, wherein the additional therapeutic agent is a triple agonist of GIP, GLP-1 and glucagon receptor. 79. The method of Example 78, wherein the triple agonist of GIP, GLP-1 and glucagon receptor is a compound comprising SEQ ID NO: 56, SEQ ID NO: 57 or SEQ ID NO: 58 or a pharmaceutically acceptable salt thereof. 80. The treatment method as described in Example 75, wherein the additional therapeutic agent is a dual agonist of glucagon and GLP-1 receptor. 81. The method of Example 80, wherein the dual agonist of glucagon and GLP-1 receptor is a compound comprising SEQ ID NO: 59, SEQ ID NO: 60, SEQ ID NO: 61 or SEQ ID NO: 62 or a pharmaceutically acceptable salt thereof. 82. The treatment method as described in Example 75, wherein the additional therapeutic agent is a agonist selective for amyloid receptors. 83. The method of Example 82, wherein the agonist selective for amyloid receptors is a compound comprising SEQ ID NO: 63, SEQ ID NO: 64, SEQ ID NO: 65 or SEQ ID NO: 66. 84. The treatment method as described in Example 75, wherein the additional therapeutic agent is a dual agonist of calcitonin receptor and amyloid receptor. 85. The method of Example 84, wherein the dual agonist of calcitonin receptor and amyloid receptor is a compound comprising SEQ ID NO: 67. 86. The treatment method as described in Example 75, wherein the additional therapeutic agent is a GLP-1 receptor agonist. 87. The method of Example 86, wherein the GLP-1 receptor agonist is a compound comprising SEQ ID NO: 79, SEQ ID NO: 80 or SEQ ID NO: 81. Example [Example] [1] [:] [Include] [SEQ ID NO: 20] [and] [SEQ ID NO: 1] [Preparation and purification of polypeptides] []

[0254] The polypeptide comprising SEQ ID NO: 20 and SEQ ID NO: 1 was prepared according to the following steps. First, the polypeptide comprising SEQ ID NO: 20 was synthesized using a ferromethoxycarbonyl (Fmoc) / tertiary butyl (t-Bu) chemical method on a Symphony 12-channel multiplex peptide synthesizer (Protein Technologies, Tucson, AZ). During solid-phase synthesis, the glass reactor vessel was wrapped with heating bands (BriskHeat controller: model SDC-120JC-A; band: model BS0051080); except for the reaction to remove the Mtt protecting group, which was carried out at ambient temperature, the heating band temperature for all reactions was set to 60°C.

[0255] Synthesized on a scale of 0.13 mmol using polystyrene Rink amide MBHA resin LL resin (Novabiochem, degree of substitution: 0.39 meq / g, 100-200 mesh, catalog number 855045). Standard side-chain protecting groups were used unless otherwise specified. For the lysine to be amide-treated, Fmoc-Lys(Mtt)-OH was used. The Fmoc group was removed using DMF containing 20% ​​piperidine (2 × 5 min) prior to each coupling step. All amino acid couplings were performed for 30 min using equimolar amounts of Fmoc amino acid (0.3 M), diisopropylcarbodiimide (0.9 M), and Oxyma (0.9 M) at an excess of 7.7 moles relative to the theoretical peptide loading. αMeL and subsequent amino acid couplings were performed for 6 h. After main-chain synthesis, the N-terminal amino acid was deprotected and subsequently mixed with DCM for 30 min and discharged. Subsequently, N-terminal acetylation was performed: a solution of 100 µL acetic anhydride, 150 µL DIPEA, and 3 mL DMF was added and mixed for 30 minutes. This step was repeated once. The following is a schematic diagram of the polypeptide containing SEQ ID NO: 20 before the addition of the linker-fatty acid moiety, using standard single-letter amino acid coding except for the N-terminal acetylation group, non-coding residues (αMeL and Aib), and C-terminal valine. The structures of these modifications and amino acid residues have been expanded.

[0256] Following N-terminal acetylation, the Mtt protecting group on the lysine to be acetylated on the resin-bound peptide was selectively removed using DCM containing 30% hexafluoroisopropanol (HFIP; Oakwood), repeated six times (6 x 15 min). Before the next step, the peptide was washed with DMF to remove residual HFIP.

[0257] For all acetylated peptides described herein, subsequent linker-fatty acid moieties were linked by performing the Fmoc / tBu chemical steps described above, to achieve the linking of amino acid monomers with standard protecting groups and the following non-standard monomers in the desired order and combination: Nα-Boc-Nε-Fmoc-L-lysine (Boc-L-Lys(Fmoc)-OH, Chem-Impex International), Nα-Fmoc-N β-Boc-L-2,3-diaminopropionic acid (Fmoc-Dap(Boc)-OH, Chem-Impex International), Fmoc-β-alanine (Fmoc-β-Ala-OH, Orpegen), Fmoc-6-aminohexanoic acid (Fmoc-Ahx-OH, described by Chem-Impex International as Fmoc-ε-Acp-OH), Fmoc-L-glutamic acid α-tert-butyl ester (Fmoc-L-Glu-OtBu, Ark Pharm), 1-(9H-furan-9-yl)-3-sideoxy-2,7,10-trioxa-4-azadodecane-12-carboxylic acid (Fmoc-AEEA-OH, ChemShuttle), mono-OtBu-octadecanoic acid (WuXi AppTec, Shanghai, China), and mono-OtBu-eicosanoic acid (WuXi... AppTec, Shanghai, China. For SEQ ID NO: 1, the order is Fmoc-Glu-OtBu, Fmoc-AEEA-OH, Boc-L-Lys(Fmoc)-OH, mono-OtBu-eicosanoic acid. Coupling for up to 6 hours was performed using 3 times the excess of reagent (equimolar amounts of AA:DIC:Oxyma).

[0258] After synthesis, the peptide resin was washed with DCM and then thoroughly air-dried. The dried resin was treated with 10 mL of lysis mixture (trifluoroacetic acid / TFA: 1,2-ethylenedithiol: triisopropylsilane: water = 89:3:3:5 v / v) for 2 hours at room temperature. The resin was collected by filtration, and the filtrate was treated with 4 times cold diethyl ether (-20°C) to precipitate the crude peptide. Subsequently, the peptide / ether suspension was centrifuged at 3200 RCF for 1.5 min to form solid aggregates. The supernatant was decanted, and the solid aggregates were ground twice more with diethyl ether. The air-dried aggregates were dissolved in acetic acid (5 mL) and water (10 mL).

[0259] The crude peptide was purified by RP-HPLC using a linear gradient of 100% acetonitrile and 0.05% TFA / water buffer on a Phenomenex phenylhexyl column (5 μm, 100A, 250×21.2, part number: 00G-4257-P0-AX). The purity of the peptide was assessed by analytical RP-HPLC using a Waters Acuity CSH C18 column (1.7 μm, 2.1×100 mm, part number 186005572), with a pooling criterion of >90%. The purity of the master pool of SEQ ID NO: 20 was found to be >92.0%. The final master pool was then lyophilized to produce the lyophilized peptide TFA salt. The molecular weight of SEQ ID NO: 20 was determined by LC / MS (experimental value: [M+3H]3+=1540.1; calculated value: [M+3H]3+=1539.1; experimental value: MW (avg)=4617.3; calculated value: MW (avg)=4614.3). The molecular weight of SEQ ID NO: 1 was determined by LC / MS (experimental value: [M+3H]3+=1781.1; calculated value: [M+3H]3+=1781.4; experimental value: MW (avg)=5340.3; calculated value: MW (avg)=5341.3). The following is a schematic diagram of the polypeptide containing SEQ ID NO: 1. Except for the N-terminal acetyl group, non-coding residues (αMeL and Aib), the acetylated lysine at position 35, and the C-terminal valine, standard single-letter amino acids are used for coding. The structures of these modifications and amino acid residues have been expanded.

[0260] A method similar to that described above and known to those skilled in the art can be used to synthesize the peptide backbone, bind the linker-fatty acid moiety, verify purity, and confirm the molecular weight of the polypeptide of the present invention described herein. The compounds of SEQ ID NO: 2 to 19, 21, 29 to 44, 46 to 48 and 51 are prepared substantially as described in the procedure of Example 1. [Example] [2] [:] [Include] [SEQ ID NO: 23] [and] [SEQ ID NO: 22] [Preparation and purification of polypeptides] []

[0261] Similar to the solid-phase synthesis in Example 1, the difference is that instead of deprotecting the N-terminal amino acid and treating it with acetic anhydride to acetylate the N-terminus, the monomer Boc-L-pyroglutamic acid (Boc-L-Pyr-OH, Combi-Blocks) is coupled under the standard conditions described in Example 1.

[0262] Similar to N-terminal Pyr preparation of sequence ID NO: 45, but using Boc-Glu-OtBu (Synthonix). Similar to N-terminal Pyr preparation of sequence IDs NO: 49 and 50, but using Boc-NMeI (Combi-Blocks).

[0263] The following is a schematic diagram of SEQ ID NO: 23. Except for the N-terminal L-Pyr-OH, non-coding residues (αMeL and Aib), and C-terminal valine, standard single-letter amino acid coding is used, wherein the structures of these modifications and amino acid residues have been expanded: The molecular weight of SEQ ID NO: 23 was determined by LC / MS (experimental value: [M+3H]3+=1582.1; calculated value: [M+3H]3+=1581.2; experimental value: MW (avg)=4743.3; calculated value: MW (avg)=4740.5).

[0264] The following is a schematic diagram of SEQ ID NO: 22. Except for the N-terminal L-Pyr-OH, non-coding residues (αMeL and Aib), the acetylated lysine at position 35, and the C-terminal valine acetylamine, standard single-letter amino acid coding is used, wherein the structure of these modifications and amino acid residues has been expanded: The molecular weight was determined by LC / MS (experimental value: [M+3H]3+=1823.4; calculated value: [M+3H]3+=1823.5; experimental value: MW (avg)=5467.2; calculated value: MW (avg)=5467.4). The compound of SEQ ID NO: 24 was prepared substantially as described in the procedure of Example 2. [Example] [3] [:] [Include] [SEQ ID NO: 27] [and] [SEQ ID NO: 25] [Preparation and purification of polypeptides] []

[0265] For this polypeptide and other polypeptides that are ultimately N-acetylated with linker-fatty acid groups, N-terminal acetylation is inappropriate; the synthesis does not include the acetic anhydride step described in Example 1. For SEQ ID NO: 27, the synthesis ends with the removal of the Fmoc protecting group on the N-terminal glycine residue.

[0266] The following is a schematic diagram of SEQ ID NO: 27. Except for the N-terminal glycine, non-coding residues (αMeL and Aib), and C-terminal valine, standard single-letter amino acid coding is used, wherein the structures of these modifications and amino acid residues have been expanded: The molecular weight of SEQ ID NO: 27 was determined by LC / MS (experimental value: [M+3H]3+=1516.9; calculated value: [M+3H]3+=1516.1; experimental value: MW (avg)=4547.7; calculated value: MW (avg)=4545.2). For SEQ ID NO: 25 and other polypeptides that are ultimately N-acylated with linker-fatty acid groups (SEQ ID NO: 26, 28, 52), Fmoc-Lys(Mtt)-OH was not used in the sequence; the synthesis also did not include the 30% HFIP deprotection step described in Example 1. For SEQ ID NO: 25, after the final Fmoc deprotection step in the synthesis of SEQ ID NO: 27, the synthesis was completed using the Fmoc / tBu chemical method as described in Example 1, wherein the added monomers were coupled / Fmoc deprotected in the following order: Fmoc-Glu-OtBu, Fmoc-AEEA-OH, Boc-L-Lys(Fmoc)-OH, mono-OtBu-eicosanoic acid.

[0267] The following is a schematic diagram of SEQ ID NO: 25. Except for the N-terminal acetylated glycine, non-coding residues (αMeL and Aib), and C-terminal valine, standard single-letter amino acids are used for coding, wherein the structures of these modifications and amino acid residues have been expanded: The molecular weight was determined by LC / MS (experimental value: [M+3H]3+ = 1758.1; calculated value: [M+3H]3+ = 1758.4; experimental value: MW (avg) = 5271.3; calculated value: MW (avg) = 5272.2). The compounds of SEQ ID NO: 26, 28, and 52 were prepared substantially as described in Example 3. [surface] [1.] The experimentally and calculated [M+3H]3+ m / z ions and MW(avg) of the polypeptides described in this paper. [SEQ ID NO] [Experimental results [M+3H], 3+ , ] [Calculated [M+3H], 3+ , ] [Experimental MW (avg)] [,g / mol)] [Calculated MW (avg)] [,g / mol)] 1 1781.0 1781.4 5340.0 5341.3 2 1781.0 1781.4 5340.0 5341.2 3 1856.5 1856.8 5566.5 5567.5 4 1776.2 1776.4 5325.6 5326.3 5 1776.2 1776.4 5325.6 5326.3 6 1966.2 1966.6 5895.6 5896.9 7 1780.5 1780.4 5338.5 5338.2 8 1804.0 1804.1 5409.0 5409.3 9 1995.8 1996.0 5984.4 5985.0 10 1795.5 1795.8 5383.5 5384.3 11 1845.9 1846.2 5534.7 5535.5 12 1773.2 1772.1 5316.6 5313.2 13 1850.9 1851.2 5549.7 5550.5 14 1830.0 1830.2 5487.0 5487.5 15 1884.3 1884.6 5649.9 5650.7 16 1916.3 1916.6 5745.9 5746.7 17 1830.6 1829.8 5488.8 5486.4 18 1819.7 1818.8 5456.1 5453.4 19 1825.1 1824.2 5472.3 5469.5 20 1540.1 1539.1 4617.3 4614.3 21 1773.1 1772.4 5316.3 5313.2 22 1823.4 1823.5 5467.2 5467.4 23 1582.1 1581.2 4743.3 4740.5 24 1815.3 1814.4 5442.9 5439.4 25 1758.1 1758.4 5271.3 5272.2 26 1758.0 1758.4 5271.0 5272.2 27 1516.9 1516.1 4547.7 4545.2 28 1750.2 1749.0 5247.6 5244.1 29 1692.8 1692.6 5075.4 5074.9 30 1686.8 1687.3 5057.4 5058.8 31 1686.9 1687.3 5057.7 5058.8 32 1686.7 1687.0 5057.1 5057.9 33 1686.6 1687.0 5056.8 5057.9 34 1686.4 1687.0 5056.2 5057.9 35 1686.6 1687.0 5056.8 5057.9 36 1686.4 1687.0 5056.2 5057.9 37 1686.6 1687.0 5056.8 5057.9 38 1681.2 1681.6 5040.6 5041.8 39 1681.3 1681.6 5040.9 5041.8 40 1681.3 1681.6 5040.9 5041.8 41 1702.9 1702.0 5105.7 5102.9 42 1707.1 1706.0 5118.3 5115.0 43 1871.8 1872.8 5612.4 5613.6 44 1686.8 1687.3 5057.4 5058.8 45 1815.9 1815.4 5444.7 5443.3 46 1785.2 1784.1 5352.6 5349.2 47 1770.1 1769.1 5307.3 5304.2 48 1770.2 1769.1 5307.6 5304.2 49 1701.6 1702.3 5101.8 5103.9 50 1657.4 1658.3 4969.2 4971.8 51 1653.2 1652.3 4956.6 4953.8 52 1744.4 1744.7 5230.2 5231.1 Example 4. In vitro assessment of CRHR1 and CRHR2 activity via intracellular cAMP production []

[0268] Corticotropin-releasing hormone (CRH) receptors 1 and 2 (CRHR1 and CRHR2) are GPCRs functionally coupled to Gs proteins. Stimulation of these receptors leads to increased intracellular cAMP production, which can be detected using standard in vitro techniques. The in vitro activity of the peptides is measured by the amount of cAMP formed in cells overexpressing human CRHR1 and CRHR2. Cell lines expressing human CRHR1 and CRHR2.

[0269] Human CRHR1 and CRHR2 receptors were stably expressed in human embryonic kidney cells (HEK293) under the control of pcDNA expression vectors. HEK293 cells were cultured in DMEM 1× (Gibco, 93-0152DK) supplemented with 10% FBS, 1 mM sodium pyruvate, 1×MEM NEAA, 1×GlutaMAX, and 10 mM HEPES. Human CRHR1-pQCXIP (T7364) and human CRHR2b-R2683puro (T16761) DNA were transfected into HEK293 cells using LipofectAMINE 3000 transfection reagent (Invitrogen, L3000001). After 20 days, mRNA levels in different pure lines were measured to confirm the expression of human CRHR1 and CRHR2 genes. To determine the function of overexpressing CRHR cells, intracellular cAMP levels in response to human UCN1 and UCN2 were measured and compared with the expression of CRHR1 and CRHR2 mRNA in various pure lines.

[0270] Human CRHR1 cells were cultured in DMEM 1× (Gibco, 93-0152DK) supplemented with 10% FBS, 1 mM sodium pyruvate, 1×MEM NEAA, 1×GlutaMAX, 10 mM HEPES, and 800 μg / ml G418. Human CRHR2 cells were cultured in the same medium (DMEM 1×, 10% FBS, 1 mM sodium pyruvate, 1×MEM NEAA, 1×GlutaMAX, 10 mM HEPES) in the presence of 0.5 μg / mL puromycin. Cells were collected after reaching 90% confluence. Cells were frozen at 10×10e6 cells / ml in a mixture of 100% fetal bovine serum and 7% DMSO and stored in liquid nitrogen until use. Natural cell lines expressing endogenous human CRHR1 and CRHR2

[0271] Y79 cells (a human retinoblastoma cell line) exhibit high levels of human CRHR1, with no detectable CRHR2. A7r5 cells (a rat aortic smooth muscle cell line) only exhibit the CRHR2 receptor, with no detectable CRHR1. The rat CRHR2 gene shares 93% nucleotide sequence homology with the human CRHR2 and responds to human UCN2. Therefore, these two natural cell lines were used for additional characterization of peptide selectivity.

[0272] Y79 cells were grown in ATCC-recommended suspension culture in a medium containing RPMI (Gibco, catalog number 11875093) and 20% FBS (Gibco dialysis type, catalog number 26400044). Y79 cells were cultured at 37°C and 5% CO2. Subcultivation was performed by removing the supernatant after centrifugation and resuspending the cell aggregates in fresh medium (subcultivation ratio 1:5). Adhesive A7r5 cells were cultured in tissue culture flasks in DMEM containing 10% FBS, 1 mM pyruvate, and 5 mM HEPES at 37°C and 5% CO2. Cultured Y79 and A7r5 cells were collected and frozen at 10 × 10e6 cells / mL in a mixture of 100% fetal bovine serum and 7% DMSO, and stored in liquid nitrogen until use. Human CRHR1 and CRHR2 cAMP analysis

[0273] On the day of analysis, frozen cells were thawed and diluted in analytical buffer (phenol red-free MEM (Corning, 17-305-CV, lot number 32121010), 0.1% casein (Sigma C4765, lot number SLCG2120), and 5 mM HEPES (pH 7.4) containing 0.25 mM IBMX). 10 μL of human CRHR1 cells (25,000 cells) or 10 μL of human CRHR2 cells (5,000 cells) were dispensed into each well of a white poly-D-lysine-coated 384-well dish (Corning, catalog number 354661). The test compound diluted in DMSO was serially added at a 1:3 ratio (200 nL / well) using an ECHO sonic liquid processor (Beckman). For hCRHR1 dishes, the mixture of cells and test compound was incubated at 37°C for 1 hour, while CRHR2 dishes were incubated at room temperature for 30 minutes.

[0274] As per the supplier's instructions, intracellular cAMP levels were quantified using HTRF (homogeneous time-varying fluorescence; Cisbio). In short, the treated cells were incubated with a lysate buffer containing 10 μL of cAMP-d2 conjugate and 10 μL of anti-cAMP-caecilin conjugate at room temperature for 60 minutes. The HTRF signal was immediately detected using an Envision disc reader (Perkin-Elmer) to calculate the fluorescence ratio from 665 to 620 nm. Human Y79 and rat A7r5 cAMP analysis

[0275] cAMP analysis of Y79 and A7r5 cells was performed similarly to the hCRHR1 and hCRHR2 analyses. Briefly, on the day of analysis, frozen cells were thawed and diluted in analysis buffer. 10 μL of human Y79 cells (5,000 cells) or 10 μL of rat A7r5 cells (5,000 cells) were dispensed into each well of a white 384-well dish. The test compound diluted in DMSO was serially added at a 1:3 ratio (200 nL / well) using an ECHO sonic liquid processor (Beckman). For both Y79 and A7r5 cells, the mixture of cells and test compounds was incubated at 37°C for 1 hour. Data Analysis

[0276] Using the cAMP standard curves generated from each experiment, the raw fluorescence data of the peptide concentration curves in Tables 2 and 3 were converted into cAMP levels (pmol / well). The relative EC50 values ​​were calculated from the upper and lower limits of the concentration-response curves using a four-parameter logic curve fitting program (GenedataScreener® v12.0.4). This curve was defined as having a maximum value for CRHR2 cells using 10 nM human UCN2 and a maximum value for CRHR1 cells using 66 nM human UCN1, with the minimum value defined only in the case of buffer. result [surface] [2.] [In humanity] [CRHR2] [and] [CRHR1] [Comparison of data on functional activity at the receptor site] [*] SEQUENCE IDS [hCRHR2 cAMP EC, 50 , ] [(pM, ± SE)] [hCRHR1 cAMP EC, 50 , (pM, ± SE)] [SEQ ID NO: 1] 9.3 ± 1.1 (n=11) >1.0e+7 (n=10) [SEQ ID NO: 2] 12.3 ± 1.8 (n=8) >1.0e+7 (n=7) [SEQ ID NO: 3] 18.3 ± 2.1 (n=3) >1.0e+7 (n=3) [SEQ ID NO: 4] 20.3 ± 2.9 (n=3) >1.0e+7 (n=3) [SEQ ID NO: 5] 10.7 ± 0.8 (n=3) >1.0e+7 (n=3) [SEQ ID NO: 6] 10.1 ± 2.5 (n=3) >1.0e+7 (n=3) [SEQ ID NO: 7] 10.6 ± 1.4 (n=3) >1.0e+7 (n=3) [SEQ ID NO: 8] 11.0 ± 0.9 (n=3) >1.0e+7 (n=3) [SEQ ID NO: 9] 15.5 ± 3.3 (n=3) >1.0e+7 (n=3) [SEQ ID NO: 10] 10.1 ± 1.1 (n=3) >1.0e+7 (n=3) [SEQ ID NO: 11] 8.3 ± 1.4 (n=4) >1.0e+7 (n=3) [SEQ ID NO: 12] 23.8 ± 1.6 (n=3) >1.0e+7 (n=3) [SEQ ID NO: 13] 7.9 ± 0.3 (n=3) >1.0e+7 (n=3) [SEQ ID NO: 14] 8.3 ± 0.5 (n=3) >1.0e+7 (n=3) [SEQ ID NO: 15] 11.3 ± 0.4 (n=3) >1.0e+7 (n=3) [SEQ ID NO: 16] 12.5 ± 1.1 (n=3) >1.0e+7 (n=3) [SEQ ID NO: 17] 9.5 ± 0.9 (n=3) >1.0e+7 (n=3) [SEQ ID NO: 18] 10.9 ± 1.3 (n=3) >1.0e+7 (n=3) [SEQ ID NO: 19] 11.1 ± 0.5 (n=3) >1.0e+7 (n=3) [SEQ ID NO: 20] 15.3 ± 1.4 (n=4) >1.0e+7 (n=4) [SEQ ID NO: 21] 16.3 ± 1.3 (n=3) >1.0e+7 (n=3) [SEQ ID NO: 22] 11.3 ± 3.0 (n=8) >1.0e+7 (n=7) [SEQ ID NO: 23] 28.9 ± 1.2 (n=3) >1.0e+7 (n=3) [SEQ ID NO: 24] 21.7 ± 3.6 (n=3) >1.0e+7 (n=3) [SEQ ID NO: 25] 6.9 ± 0.5 (n=12) >1.0e+7 (n=11) [SEQ ID NO: 26] 6.9 ± 0.7 (n=3) >1.0e+7 (n=3) [SEQ ID NO: 27] 23.6 ± 2.9 (n=3) >1.0e+7 (n=3) [SEQ ID NO: 28] 12.8 ± 2.0 (n=3) >1.0e+7 (n=3) [SEQ ID NO: 29] 5.6 ± 0.4 (n=17) >1.0e+7 (n=11) [SEQ ID NO: 30] 5.7 ± 0.1 (n=4) >1.0e+7 (n=3) [SEQ ID NO: 31] 4.5 ± 0.7 (n=4) >1.0e+7 (n=3) [SEQ ID NO: 32] 5.4 ± 0.7 (n=5) >1.0e+7 (n=3) [SEQ ID NO: 33] 4.6 ± 0.6 (n=5) 1.1e+6 ± 6.1e+4 (n=3) [SEQ ID NO: 34] 4.9 ± 0.7 (n=5) >1.0e+7 (n=3) [SEQ ID NO: 35] 5.3 ± 0.9 (n=5) 3.0e+6 ± 4.6e+5 (n=3) [SEQ ID NO: 36] 4.4 ± 0.4 (n=6) 1.8e+6 ± 3.2e+5 (n=3 / 4) [SEQ ID NO: 37] 5.0 ± 0.2 (n=6) 1.7e+6 ± 2.8e+5 (n=3) [SEQ ID NO: 38] 6.3± 2.1 (n=4) 1.1e+6 ± 2.9e+5 (n=2) [SEQ ID NO: 39] 7.3 ± 0.8 (n=4) 3.7e+6 (n=1 / 2) [SEQ ID NO: 40] 10.3 ± 1.9 (n=4) >1.0e+7 (n=1 / 2) [SEQ ID NO: 41] 7.0 ± 0.9 (n=7) 3.7e+6 ± 3.1e+5 (n=6) [SEQ ID NO: 42] 6.6 ± 0.8 (n=10) 2.3e+6 ± 1.9e+5 (n=9) [SEQ ID NO: 43] 7.9 ± 1.3 (n=3) >1.0e+7 (n=3) [SEQ ID NO: 44] 5.4 ± 0.5 (n=4) >1.0e+7 (n=3) [SEQ ID NO: 45] 9.0 ± 3.6 (n=3) 3.7e+6 ± 5e+5 (n=3) [SEQ ID NO: 46] 13.0 ± 1.1 (n=9) >1.0e+7 (n=7) [SEQ ID NO: 47] 17.5 ± 2.2 (n=9) 2.3e+6 ± 1.3e+5 (n=3 / 8) [SEQ ID NO: 48] 26.7 ± 3.8 (n=10) >1.0e+7 (n=9) [SEQ ID NO: 49] 6.6 ± 0.9 (n=19) >1.0e+7 (n=7) [SEQ ID NO: 50] 22.4 ± 3.6 (n=24) >1.0e+7 (n=14) [SEQ ID NO: 51] 8.6 ± 3.2 (n=3) 4.3e+6 ± 1.4e+5 (n=3) [SEQ ID NO: 52] 6.6 ± 0.5 (n=3) 1.5e+6 (n=1 / 3) [SEQ ID NO: 53] 24.5 ± 1.6 (n=124) >1.0e+7 (n=65) [SEQ ID NO: 54] 29.8 ± 2.5 (n=127) >1.0e+7 (n=64) This table shows that all the peptides of this invention have specific activity at the hCRHR2 receptor and almost no activity at the hCRHR1 receptor. [surface] [3.] [Human] [Y79] [and rats] [A7r5] [Comparison of data on functional activities in cells] [] SEQ ID A7r5 cAMP EC50 (pM, ± SE) Y79 cAMP EC50 (pM, ± SE) SEQ ID NO: 1 157 ± 8 (n=5) 1.4e+6 ± 2.8e+5 (n=3) SEQ ID NO: 2 544 ± 133 (n=2) 2.1e+6 ± 3.3e+5 (n=2) SEQ ID NO: 22 314 ± 24 (n=5) 2.4e+6 ± 3.1e+5 (n=3) SEQ ID NO: 25 112 ± 5 (n=5) 6.2e+5 ± 1.8e+5 (n=3) SEQ ID NO: 29 123 ± 2 (n=4) 1.0e+6 ± 1.1e+5 (n=2) SEQ ID NO: 46 313 ± 24 (n=4) 4.0e+6 ± 4.5e+5 (n=2) SEQ ID NO: 47 1061 ± 176 (n=2) 7.2e+5 ± 3.0e+5 (n=2) SEQ ID NO: 48 1156 ± 29 (n=2) 4.5e+6 ± 1.4e+5 (n=2) SEQ ID NO: 49 137 ± 14 (n=4) >1e+7 (n=2) SEQ ID NO: 50 967 ± 347 (n=4) >1e+7 (n=2) SEQ ID NO: 53 102 ± 11 (n=5) >1e+7 (n=3) SEQ ID NO: 54 471 ± 85 (n=5) 6.0e+6 ± 2.6e+6 (n=3) Similar to the peptide selectivity observed in hCRHR2 cells relative to hCRHR1 cells, the data presented in this table indicate that all peptides of the present invention exhibit significantly higher efficacy in A7r5 cells (CRHR2 receptors) than in Y79 cells (CRHR1 receptors). [Example] [5] [:] [Using settling velocity analysis-based ultracentrifugation] [(SV-AUC)] [Determination of association status] [, , ]

[0277] SV-AUC was performed at 20°C using a Beckman Optima with an AN 60 Ti rotor at a final rotor speed of 60,000 rpm. Peptides were dialyzed into 5 mM phosphate (pH 7) or 1× phosphate-buffered saline (PBS) (pH 7.2). Samples were adjusted to the desired concentration (0.1 mg / mL, 1 mg / mL, or 10 mg / mL) using the appropriate buffer as needed. Both samples and buffers were filtered through a 0.22 μm PVDF centrifuge filter unit. The filtered samples were loaded into the sample area of ​​the cell; only the filtered buffer was loaded into the reference area of ​​the cell. Wavelength scanning was performed as the first step to select a suitable λ value such that the OD of a given compound at the test concentration was between 1 and 1.2; this value was typically between 225 nm and 254 nm and was used for the method scan in the next step. The λ values ​​for each compound are shown in Table 1. Both absorbance and interference detection modules were used in the method scan. Temperature equilibration was set to the first stage at a rotor speed of 0 rpm for 3 hours. Settling velocity was set to the second stage at a rotor speed of 60,000 rpm. At a specified wavelength, 300 scans were performed at a frequency of 60 seconds, or 500 scans were performed at a frequency of 120 seconds. Data was analyzed using SEDFIT software, employing a continuous c(s) distribution model; relevant parameter settings are shown in Tables 4 and 5; settling coefficient (s) values ​​are reported in Svedherg units (S). [, , ] [] [surface] [4.] [Overview of compound concentration, buffer solution, wavelength, sedimentation coefficient, molecular weight, and apparent molecular weight] [] SEQ ID NO. Concentration (mg / mL) buffer solution λ (nm) Settlement coefficient (percentage of total) MW (g / mol) Mw (SEDFIT) 54 0.1 1×PBS, pH 7.2 225 3.5 S (89%) 5157.1 38787 Da 1 0.9 S (5%), 2.5 S (2.5%), 3.7 S (88.5%) 5608 Da, 27636 Da, 50313 Da 10 0.9 S (25.3%), 2.9 S (12.4%), 3.9 S (51.2%) 5654 Da, 34479 Da, 53507 Da 49 10 1×PBS, pH 7.2 252<00​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​0.8 S (11.8%) 1.4 S (89.3%) 5304.3 7027 Da, 17800 Da 48 1 5 mM phosphate, pH 7 238 0.9 S (11%), 1.5 S (87.2%) 5304.3 7278 Da, 16252 Da 2 1 5 mM phosphate, pH 7 237 0.9 S (10%) 1.5 S (84.6%) 5341.3 8092 Da, 17142 Da 1 1 5 mM phosphate, pH 7 234 0.9 S (13.7%) 1.5 S (87.5%) 5341.3 1 1 5 mM phosphate, pH 7 + 150 mM NaCl 234 1.25 S (8.9%) 1.82 S (90.2%) 5341.3 twenty two 1 5 mM phosphate, pH 7 237 0.8 S (13.3%) 1.5 S (86.4%) 5467.4 twenty two 1 5 mM phosphate, pH 7 + 150 mM NaCl 237 1.33 S (8.3%) 1.82 S (90.6%) 5467.4 25 1 5 mM phosphate, pH 7 237 0.7 S (8.6%) 1.4 S (87.3%) 5272.2 25 1 5 mM phosphate, pH 7 + 150 mM NaCl 237 0.7 S (2.9%) 1.75 S (85.4%) 5272.2 The experiments characterizing the compound containing SEQ ID NO: 54 were performed at 300 scan counts over 60 seconds; for all other test peptides, the experiments were performed at 500 scan counts over 120 seconds. [surface] [5.] [Use Continuous] [c(s)] [Distribution model for] [SEDFIT] [Software settings related parameters] [.] (a) Parameters used for the continuous settlement coefficient distribution c(s) of SEQ ID NO: 54 [parameter] [value] Resolution 50 minimum s 0.0 Maximum s 20.0 Friction ratio 1.2 Baseline 0.00000 Fitting RI noise yes Fitting time-independent noise no meniscus 5.8002 bottom Unfixed Trust level (F ratio) 0.67000 Partial volume 0.73000 Buffer density 1.00000 Buffer viscosity (poise) 0.01002 Obtain the S-gradient from the file no Using logarithmic intervals in an s-grid no (b) Parameters for other peptides used in this invention [parameter] [value] Resolution 50 minimum s 0.0 Maximum s 5.0 Friction ratio 1.2 Baseline 0.00000 Fitting RI noise yes Fitting time-independent noise yes meniscus 6.1357 bottom Unfixed Trust level (F ratio) 0.95000 Partial volume 0.73000 Buffer density 1.00000 Buffer viscosity (poise) 0.01002 Obtain the S-gradient from the file no Using logarithmic intervals in an s-grid no

[0278] Referring to the sedimentation coefficient of compound SEQ ID NO: 54, the higher oligomers were the dominant species at all three tested concentrations (0.1 mg / mL, 1 mg / mL, and 10 mg / mL). In contrast, the peptides of this invention were measured to be in a lower self-associative state compared to SEQ ID NO: 54. [Example] [6.] [In vivo diet-induced obesity] [(DIO)] [Model] [-] [Long-term dosage] [, , ]

[0279] The DIO model represents a prediabetic state, i.e., obesity. These animals, though not yet diabetic, exhibited insulin resistance, dyslipidemia, and hepatic steatosis after 12 weeks on a high-fat diet (60% of kcal from fat), all characteristic of metabolic syndrome (Surwit RS et al., Diet-induced type II diabetes in C57BL / 6J mice. Diabetes 37(9): 1163-7 (1988)). The assessment of changes in body weight, fat mass, and lean mass since the start of the study, as well as changes in fasting glucose or glucose compared to the carrier in the same study, is essentially as described below. In the case of the combination of SEQ ID NO: 55 and SEQ ID NO: 25, the tibialis anterior muscle weight at the end of the study was also reported as a measure of the effect on muscle weight.

[0280] One male C57BL6 mouse aged 22–24 weeks was housed in each cage (on a high-fat diet from 6 weeks of age, Jackson Laboratories 3800050; Bar Harbor, ME) and maintained in the animal house for two weeks prior to the start of the experiment on D12492 diet (60% lard high-fat diet: Research diets New Brunswick NJ) with a normal light cycle. Animals were randomly assigned to the treatment group based on body weight using group randomization. On day 1 of the experiment, animals and food were weighed and recorded. Animals were divided into two identical groups and started on different dates (data merged) to simplify research logistics. On days 1 (start), 4, 7, 10, and 13 of the experiment, animals were given a single subcutaneous injection (sc) at a volume of 10 ml / kg of the treatment agent specified in 40 mM Tris HCl pH 8.0 + 50 mM D-mannitol + 0.02% polysorbate 80. The mediator control animals were injected with a similar volume of this solution. Throughout the study period, the solution was stored in sterile capped vials at 4°C. Each treatment arm contained n=5 to 6 mice per group. weight

[0281] From day 1 to day 14 of the study, animals were weighed daily before dose administration. This data was used to calculate changes in body weight. Animals were placed in the weighing pan and allowed to stabilize on the balance. The weight was recorded. Fasting glucose

[0282] On day 14 of the study, animals were placed in clean cages with clean wire mesh, fasted overnight (approximately 16 to 18 hours) without food but allowed to drink water. On day 15, fasting blood glucose was measured in the animals as follows: tails were removed, blood samples were collected, and blood glucose was measured using an Accu-Chek Aviva glucose meter (Roche; Indianapolis, IN). Statistical significance was calculated using GraphPad Prizm software (La Jolla, Ca) (*=p>0.05, relative to 0 dose; one-way ANOVA, Dunnett post-hoc analysis). Body composition analysis

[0283] Body composition was analyzed using a quantitative nuclear magnetic resonance (EMR-166-s, EchoMRI, Houston Tx) analyzer on days 1 and 14 of the study. After calibrating the analyzer with a known amount of rapeseed oil, animals were placed in the analyzer, which measures fat and lean body mass in grams. Changes in mass were calculated by subtracting the day 14 value from the day 1 value. Tibialis anterior muscle weight

[0284] On day 15 of the study, after measuring blood glucose levels by tail sampling, the animals were euthanized by CO2 asphyxiation, and the right tibialis anterior muscle was dissected. The muscle fascia and tendon were removed, and the muscle was weighed. The weight was recorded, and in the case of relative weight, the weight of the tibialis anterior muscle (in milligrams) was divided by the body weight (in grams).

[0285] Tables 6 through 11 below show the data corresponding to the measurements above. Data are expressed as the arithmetic mean plus or minus the standard error (SEM) of the mean. Tables 6 through 9 show the ED 50 for the listed parameters as needed. ED 50 was calculated using the Logarithm (Accelerator) vs. Response – Find ECanything function in GraphPad Prism software (La Jolla, Ca), where F=50 and the upper limit (top) is fixed at the mediator mean. [surface] [6] [:] [Include] [SEQ ID NO: 29] [Dose-response data for the polypeptide] [] [0 nmol / kg (n=5)] [0.3 nmol / kg (n=6)] [1 nmol / kg (n=6)] [3 nmol / kg n=6] [10 nmol / kg (n=6)] [30 nmol / kg (n=6)] [100 nmol / kg (n=6)] [300 nmol / kg (n=6)] [ED 50 (nmol / kg)] Weight change (g) 1.5 ±0.21 0.88 ±0.57 0.30 ±0.36 -0.72 ±0.38* -5.43 ±0.54* -6.51 ±0.75* -8.12 ±0.40* -10.42 ±0.65* 10.56 Changes in fat mass (g) 1.20 ±0.23 1.09 ±0.86 -1.33 ±.041 -2.40 ±0.68* -5.76 ±0.43* -5.86 ±0.93* -7.57 ±2.20* -9.29 ±0.45* 6.62 Changes in lean body mass (g) -0.52 ±0.46 0.43 ±0.74 0.70 ±0.37 1.76 ±0.62 0.64 ±0.39 -0.80 ±0.63 -0.56 ±0.82 -1.30 ±0.55 Unclear Fasting blood glucose (mg / dL) 129.6 ±10.14 111.7 ±5.04 95.78 ±3.28* 91.67 ±3.09* 89.28 ±1.70* 92.61 ±2.50* 90.22 ±3.50* 84.67 ±3.10* 0.35 *=p<0.05, relative to the mediator (one-way ANOVA, Dunnett post-hoc analysis) [surface] [7] [:] [Include] [SEQ ID NO: 1] [Dose-response data for the polypeptide] [] [0 nmol / kg(n=6)] [0.3 nmol / kg (n=6)] [1 nmol / kg (n=6)] [3 nmol / kg (n=6)] [10 nmol / kg (n=6)] [30 nmol / kg (n=6)] [100 nmol / kg (n=6)] [300 nmol / kg(n=6)] [ED 50 (nmol / kg)] Weight change (g) 0.85 ±0.79 1.63 ±0.59 -0.05 ±0.7 -1.27 ±0.72 -4.23 ±0.85* -7.05 ±0.83* -9.60 ±0.63* -9.50 ±0.78* 11.27 Changes in fat mass (g) 0.44 ±0.63 -0.23 ±0.45 -1.69 ±0.67 -3.02 ±0.71* -5.56 ±0.52* -7.19 ±0.66* -9.20 ±0.53* -8.97 ±0.55* 6.45 Changes in lean body mass (g) -0.19 ±0.30 1.09 ±0.10* 0.86 ±0.30 1.42 ±0.20* 1.00 ±0.46* 0.47 ±0.24 0.15 ±0.27 -0.62 ±0.32 Unclear Fasting blood glucose (mg / dL) 132.2 ±4.79 115.8 ±5.43 97.00 ±4.50* 90.33 ±2.89* 86.00 ±3.96* 89.00 ±1.53* 86.40 ±5.54* 84.44 ±2.22* 0.45 *=p<0.05, relative to the mediator (one-way ANOVA, Dunnett post-hoc analysis) [surface] [8] [:] [Include] [SEQ ID NO: 22] [Dose-response data for the polypeptide] [] [0 nmol / kg] [0.3 nmol / kg] [1 nmol / kg] [3 nmol / kg] [10 nmol / kg] [30 nmol / kg] [100 nmol / kg] [300 nmol / kg] [ED 50 (nmol / kg)] Weight change (g) 1.23 ±0.49 0.15 ±0.51 0.67 ±0.34 -1.87 ±0.71* -4.75 ±0.52* -8.82 ±0.48* -9.72 ±0.62* -12.27 ±0.84* 12.50 Changes in fat mass (g) 0.93 ±0.37 -0.92 ±0.35* -0.96 ±0.35* -2.90 ±0.51* -5.56 ±0.45* -8.34 ±0.31* -8.86 ±0.52* -10.95 ±0.50* 9.80 Changes in lean body mass (g) -0.96 ±0.24 -0.03 ±0.08 0.42 ±0.14* 0.47 ±0.21* 0.11 ±0.16* -0.56 ±0.29 -0.67 ±0.12 -0.91 ±0.35 Unclear Fasting blood glucose (mg / dL) 162.4 ±9.54 105.70 ±5.74 105.20 ±6.01 93.92 ±2.91 93.50 ±4.89* 87.42 ±3.17 91.58 ±2.14* 89.25 ±2.90* 0.03 *=p<0.05, relative to the mediator (one-way ANOVA, Dunnett post-hoc analysis) [, , ] [] [surface] [9] [:] [Include] [SEQ ID NO: 25] [Dose-response data for the polypeptide] [] [0 nmol / kg (n=6)] [0.3 nmol / kg (n=6)] [1 nmol / kg (n=6)] [3 nmol / kg (n=6)] [10 nmol / kg (n=6)] [30 nmol / kg (n=6)] [100 nmol / kg (n=6)] [300 nmol / kg(n=6)] [ED 50 (nmol / kg)] Weight change (g) 1.78 ±0.72 1.47 ±1.39 -0.62 ±0.47 0.33 ±0.97 -5.03 ±0.68* -6.67 ±0.53* -9.07 ±0.76* -10.10 ±0.74* 11.11 Changes in fat mass (g) 1.15 ±0.40 0.05 ±1.06 -2.01 ±0.49* -1.60 ±0.69* -5.45 ±0.26* -6.71 ±0.49* -8.57 ±0.50* -9.16 ±0.64* 8.46 Changes in lean body mass (g) 0.04 ±0.26 0.85 ±0.32 0.81 ±0.28 1.19±0.37 0.57 ±0.64 0.36 ±0.17 -0.16 ±0.28 -0.46 ±0.10 Unclear Fasting blood glucose (mg / dL) 146.20 ±13.74 113.6 ±5.14* 105.60 ±6.32* 92.67 ±2.94* 90.33 ±3.77* 90.42 ±1.70* 86.58 ±3.18* 87.08 ±2.86* 0.25 *=p<0.05, relative to the mediator (one-way ANOVA, Dunnett post-hoc analysis) [surface]

[10] [:] [Include] [SEQ ID NO: 50] [、] [SEQ ID NO: 41] [、] [SEQ ID NO: 42] [、] [SEQ ID NO: 45] [、] [SEQ ID NO: 46] [、] [SEQ ID NO: 47] [、] [SEQ ID NO: 2] [and reference compounds] [SEQ ID NO: 54] [Single dose of the polypeptide] [(30 nmol / kg)] [material] [] [SEQ ID NO: 50 30 nmol / kg] [SEQ ID NO: 41 30 nmol / kg] [SEQ ID NO: 42] [30 nmol / kg] [SEQ ID NO: 45 30 nmol / kg] [SEQ ID NO: 46 30 nmol / kg] [SEQ ID NO: 47] [30 nmol / kg] [SEQ ID NO: 48 30 nmol / kg] [SEDQ ID NO:2 30 nmol / kg] [SEQ ID NO: 54 30 nmol / kg (] [Combined data from four studies] Weight change (g) -7.20 ±0.57* -6.42 ±0.70* -4.88 ±0.45* -4.93 ±0.80* -5.80 ±0.62* -4.87 ±0.68* -6.16 ±0.65* -4.87 ±0.68* -3.75 ±0.32 Changes in fat mass (g) -7.18 ±0.47* -6.59 ±0.54* -5.33 ±0.40* -5.40 ±0.63* -5.63 ±0.42* -4.80 ±0.54* -5.73 ±0.41* -5.76 ±0.96* -4.05 ±0.22 Changes in lean body mass (g) -0.02 ±0.16 0.40 ±0.23 0.48 ±0.13 0.56 ±0.16 -0.19 ±0.45 0.26 ±0.45 -0.19 ±0.37 -0.35 ±0.56 0.60 ±0.17 Fasting blood glucose (reduced relative to the drug) % 40.16 ±3.15* 42.83 ±3.28* 29.41 ±3.43* 40.88 ±1.70* 35.44 ±1.98* 27.62 ±4.06* 32.53 ±3.35* 37.33 ±1.68* 37.01 ±1.50 *=p<0.05, relative to mediators in the same study (one-way ANOVA, Dunnett post-hoc analysis) [surface]

[11] [:] [Include] [SEQ ID NO: 55] [The compound alone and with inclusions] [SEQ ID NO: 25] [Single dose of a series of dosage combinations of compounds] [(10 nmol / kg)] [Information] [] [SEQ ID NO: 55] [0 nmol / kg] [10 nmol / kg] [10 nmol / kg] [10 nmol / kg] [10 nmol / kg] [10 nmol / kg] [10 nmol / kg] [10 nmol / kg] [10 nmol / kg] [SEQ ID NO: 25] [0 nmol / kg (n=6)] [0 nmol / kg (n=6)] [0.3 nmol / kg (n=6)] [1 nmol / kg (n=6)] [3 nmol / kg (n=6)] [10 nmol / kg (n=6)] [30 nmol / kg (n=6)] [100 nmol / kg (n=6)] [300 nmol / kg (n=6)] Weight change (g) 1.23 ±0.49 -10.33 ±0.86* -9.43 ±0.74* -12.45 ±1.25* -11.15 ±0.54* -12.55 ±0.66* -14.77 ±0.93*# -16.37 ±1.36*# -17.08 ±0.89*# Changes in fat mass (g) 0.93 ±0.37 -7.55 ±0.63* -7.61 ±0.73* -10.45 ±0.98*# -9.60 ±0.49* -11.14 ±0.55*# -12.32 ±0.72*# -13.36 ±0.72*# -13.55 ±0.65*# Changes in lean body mass (g) -0.96 ±0.24 -2.31 ±0.31 -1.74 ±0.16 -1.81 ±0.21 -1.42 ±0.28 -1.07 ±0.24 -2.02 ±0.31 -2.87 ±0.65* -3.02 ±0.35* Fasting blood glucose (mg / dL) 162.4 ±9.54 77.67 ±3.52* 82.33 ±2.36* 76.83 ±2.95* 76.08 ±6.31* 81.25 ±4.57* 76.50 ±2.75* 71.08 ±5.54* 79.42 ±2.00* Absolute shin weight (mg) 70.32 ±2.72 55.45 ±3.21 68.95 ±3.90 75.88 ±3.68# 78.32 ±5.10# 80.72 ±6.63# 83.23 ±5.49# 76.22 ±6.86# 69.72 ±2.45 Relative shin weight (mg / g body weight) 1.49 ±0.06 1.62 ±0.15 1.92 ±0.12 2.33 ±0.16*# 2.30 ±0.15*# 2.49 ±0.16*# 2.76 ±0.19*# 2.63 ±0.14*# 2.55 ±0.18*# * = p < 0.05, relative to the mediator; # = p < 0.05, relative to SEQ ID NO: 55 Tirzepatide 10 nmol / kg (single-factor ANOVA, Dunnett post-hoc analysis)

[0286] When the test peptide was injected subcutaneously every three days for 14 consecutive days, the following significant differences were observed: (1) weight loss, (2) reduction in fat mass, and (3) decrease in fasting blood glucose, and (4) no significant change in lean body mass. Dose-response experiments with SEQ ID NO: 29, SEQ ID NO: 1, SEQ ID NO: 22 and SEQ ID NO: 25 showed that, in terms of (1) changes in weight and (2) changes in fat mass measured by ED 50, their efficacy was improved compared to the previous peptide SEQ ID NO: 54 (Tables 6 to 9). At a single dose of 30 nmol / kg, the peptides containing SEQ ID NO: 50, SEQ ID NO: 41, SEQ ID NO: 42, SEQ ID NO: 45, SEQ ID NO: 46, SEQ ID NO: 47, SEQ ID NO: 48, and SEQ ID NO: 2 also showed superior efficacy to the previously mentioned peptide SEQ ID NO: 54 in terms of (1) changes in body weight and (2) changes in fat mass (Table 10). As shown in Table 11, when combined with SEQ ID NO: 55 at different doses, the peptide containing SEQ ID NO: 25 enhanced the weight loss and fat mass reduction induced by SEQ ID NO: 55, and statistically significant differences were observed at doses as low as 30 nmol / kg SEQ ID NO: 25 (in terms of changes in total body weight) and 1 nmol / kg SEQ ID NO: 25 (in terms of changes in fat mass). Compared to 10 nmol / kg SEQ ID NO: 55 alone, when doses as low as 1 nmol / kg are combined with 10 nmol / kg SEQ ID NO: 55, the peptide containing SEQ ID NO: 25 also increases the relative and absolute weight of the tibialis anterior muscle (Table 11). [Example] [7] [:] [Pharmacokinetics] [, , ] Bioanalytical methods

[0287] Plasma concentrations of the compounds were determined by LC / MS. For each analysis, the compounds and internal standards (IS) were extracted from 100% mouse, rat, pig, or monkey plasma (70 µL) using [50-50]isopropanol-methanol. After centrifugation, two distinct layers were formed, with the compounds and IS in the supernatant layer. Aliquots (150 µL) of the supernatant were transferred to a Waters Sirocco protein precipitation plate. The supernatant was dissociated using a positive pressure manifold. The supernatant was mixed with 100-2 water-formic acid (400 µL) in a pretreated Sep-Pak tC18 SPE plate and dissociated into waste. The peptides and IS were dissociated in two separate steps (50 µL each) using [100-2]acetonitrile-formic acid (100 µL). The elution solution (40 µL) from the SPE disk was mixed with dilution solutions (1.875 µL of 5×invitrosol and 3.125 µL of 1% formic acid aqueous solution, 80 µL). The final sample (10 µL) was loaded onto an Xselect CSH C18 IS column (3.5 µm, 2.1 × 20 mm). The column effluent was guided to a Thermo Fisher Scientific Orbitrap (Explororis 240) mass spectrometer for detection and quantification. Pharmacokinetic studies

[0288] A single subcutaneous or intravenous dose (183 to 190 nmol / kg) of the compound in 40 mM Tris-HCl 5% D-mannitol (pH 8) was administered to male and female rock crab macaques at a volume of 0.5 mL / kg. Blood samples were collected from each animal at 0.5, 3, 6, 24, 48, 72, 96, 168, 240, 336, 408, and 504 hours after administration for pharmacokinetic characterization.

[0289] A single subcutaneous or intravenous dose (19 or 99 nmol / kg) of the compound in 40 mM Tris-HCl 5% D-mannitol (pH 8) was administered to male Yucatan miniature pigs at a volume of 0.01 or 0.05 mL / kg. Blood samples were collected from each animal at 0.083 (IV only), 0.5, 3, 6, 24, 48, 72, 96, 168, 240, 336, 408, and 504 hours after administration for pharmacokinetic characterization.

[0290] A single subcutaneous dose (183 to 201 nmol / kg) of the compound in 40 mM Tris-HCl 5% D-mannitol (pH 8) was administered to male Sprague Dawley rats at a volume of 0.5 or 2 mL / kg. Blood samples were collected from each animal at 0.5, 3, 6, 24, 48, 72, 96, 120, and 168 hours post-administration for pharmacokinetics characterization.

[0291] A single subcutaneous dose (10, 30, or 100 nmol / kg) of the compound in 40 mM Tris-HCl 5% D-mannitol (pH 8) was administered to male Sporgodoryl rats at a volume of 1 mL / kg. Blood samples were collected from each animal at 3, 6, 24, 48, 72, 96, 120, and 168 hours post-administration for pharmacokinetics characterization.

[0292] A single subcutaneous dose (100 nmol / kg) of the compound in 40 mM Tris-HCl 5% D-mannitol (pH 8) was administered to male CD-1 rats at a volume of 5 mL / kg. Blood samples were collected at 6, 24, 48, 96, 168, 240, 288, and 336 hours post-administration for pharmacokinetic characterization. [surface]

[12] [:] [Individual and mean pharmacokinetic parameters of male and female stone crab macaques after a single subcutaneous dose] [SEQ ID] [(] [dose)] [Animal_ID] [T, 1 / 2 , ] [Tmax] [Cmax] [AUC, 0-inf , ] [CL / F] [(hr)] [(hr)] [(nmole / L)] [(hr*nmole / L)] [(mL / hr / kg)] SEQ ID NO: 1 (187 nmol / kg) PM214419 72.0 48 1100 221000 0.867 PM220568 91.7 48 1260 255000 0.755 average value 81.9 48 1180 238000 0.811 SEQ ID NO: 22 (183 nmol / kg) PM220212 182 72 1720 387000 0.522 PM220229 90.2 48 2280 342000 0.552 average value 136 60 2000 365000 0.537 SEQ ID NO: 25 (190 nmol / kg) PM220563 184 twenty four 1540 364000 0.599 PM220223 214 twenty four 1680 551000 0.423 average value 199 twenty four 1610 458000 0.511 Abbreviations: AUC0-inf = Area under the curve from 0 to infinity, CL / F = Clearance / Bioavailability, Tmax = Time to reach maximum concentration, Cmax = Maximum observed plasma concentration, T1 / 2 = Half-life. [surface]

[13] [:] [Individual and mean pharmacokinetic parameters of male and female stone crab macaques after a single intravenous dose] [] [SEQ ID] [(] [dose)] [Animal_ID] [T, 1 / 2 , ] [C, 0, ] [AUC, 0-inf , ] [CL] [(hr)] [(nmole / L)] [(hr*nmole / L)] [(mL / hr / kg)] SEQ ID NO: 1 (187 nmol / kg) PM222982 91.3 4150 261000 0.717 PM203046 91.9 5040 369000 0.507 average value 91.6 4600 315000 0.612 SEQ ID NO: 22 (183 nmol / kg) PM210594 83.1 4160 467000 0.392 PM210588 113 5320 424000 0.431 average value 98.1 4740 446000 0.412 SEQ ID NO: 25 (190 nmol / kg) PM214424 100 3040 355000 0.534 PM220569 126 3960 449000 0.422 average value 113 3500 402000 0.478 Abbreviations: AUC0-inf = Area under the curve from time 0 to infinity, CL = clearance rate, C0 = estimated plasma concentration at time 0, T1 / 2 = half-life. [surface]

[14] [:] [Individual and mean pharmacokinetic parameters of male Yucatan pigs after a single subcutaneous dose] [] [SEQ ID NO:] [(] [dose)] [Animal_ID] [T, 1 / 2 , ] [Tmax] [Cmax] [AUC, 0-inf , ] [CL / F] [(hr)] [(hr)] [(nmole / L)] [(hr*nmole / L)] [(mL / hr / kg)] SEQ ID NO: 29 (99 nmol / kg) 2001 166 twenty four 497 128000 1.03 2002 292 twenty four 201 67500 2.06 2003 185 168 79.6 33800 3.56 average value 214 72 259 76400 2.22 SD 67.9 83 215 47700 1.27 SEQ ID NO: 48 (19 nmol / kg) 1519 70.1 48 140 29300 0.653 1523 66.8 6.0 157 35600 0.565 average value 68.5 27 149 32500 0.609 SEQ ID NO: 2 (19 nmol / kg) 1578 70.7 6 154 24400 0.788 1857 90.8 96 60.4 11700 1.63 average value 80.8 51 107 18100 1.21 SEQ ID NO: 1 (19 nmol / kg) 1572 91.6 6 171 22400 0.848 1579 60.2 6 122 13100 1.46 average value 75.9 6 147 17800 1.15 SEQ ID NO: 22 (18 nmol / kg) 1944 79.1 twenty four 165 23900 0.777 1947 85.8 6.0 143 21000 0.886 average value 82.5 15 154 22500 0.832 SEQ ID NO: 25 (19 nmol / kg) 1954 80.8 twenty four 159 28300 0.681 1988 70.4 twenty four 181 26800 0.717 average value 75.6 twenty four 170 27600 0.699 Abbreviations: AUC0-inf = Area under the curve from 0 to infinity, CL / F = Clearance / Bioavailability, Tmax = Time to reach maximum concentration, Cmax = Maximum observed plasma concentration, T1 / 2 = Half-life. [surface]

[15] [:] [Individual and mean pharmacokinetic parameters of male Yucatan pigs after a single intravenous dose] [] [Compound] [(] [dose)] [Animal_ID] [T, 1 / 2 , ] [C, 0, ] [AUC, 0-inf , ] [CL] [(hr)] [(nmole / L)] [(hr*nmole / L)] [(mL / hr / kg)] SEQ ID NO: 29 (99 nmol / kg) 1001 126 2560 220000 0.448 1002 263 2380 329000 0.299 1003 109 2530 307000 0.321 average value 166 2490 285000 0.356 SD 84.4 96.4 57600 0.080 SEQ ID NO: 48 (19 nmol / kg) 1570 86.7 899 32500 0.580 2711 89.2 987 41200 0.458 average value 88.0 943 36900 0.519 SEQ ID NO: 2 (19 nmol / kg) 1530 95.4 809 28900 0.648 1568 75.1 578 32800 0.571 average value 85.3 694 30900 0.610 SEQ ID NO: 1 (19 nmol / kg) 1561 87.0 591 32100 0.583 1565 55.9 646 20200 0.927 average value 71.5 619 26200 0.755 SEQ ID NO: 22 (18 nmol / kg) 1933 90.1 373 22300 0.820 1938 101 462 28300 0.646 average value 95.6 418 25300 0.733 SEQ ID NO: 25 (19 nmol / kg) 1950 106 522 41900 0.453 1963 86.8 545 37200 0.510 average value 96.4 534 39600 0.482 Abbreviation: AUC0-inf = area under the curve from time 0 to infinity, CL = clearance rate, C0 = plasma concentration estimated at time 0, T1 / 2 = half-life. [surface]

[16] [:] [Individual and mean pharmacokinetic parameters of male Sporgodory rats after a single subcutaneous dose] [] [Compound] [(] [dose)] [Animal_ID] [T, 1 / 2 , ] [Tmax] [Cmax] [AUC, 0-inf , ] [CL / F] [(hr)] [(hr)] [(nmole / L)] [(hr*nmole / L)] [(mL / hr / kg)] SEQ ID NO: 49 (196 nmol / kg) R0101 19.1 twenty four 340 17100 11.5 R0102 16.0 6 474 22400 8.76 R0103 20.2 twenty four 367 17300 11.4 average value 18.4 18 394 18900 10.6 SD 2.18 10 70.9 3000 1.55 SEQ ID NO: 50 (201 nmol / kg) R0301 19.7 twenty four 577 26700 7.55 R0302 14.9 twenty four 460 25100 8.03 R0303 16.7 twenty four 618 28700 7.01 average value 17.1 twenty four 552 26800 7.53 SD 2.42 0.0 82.0 1800 0.510 SEQ ID NO: 29 (197 nmol / kg) R0501 52.8 twenty four 881 96800 2.35 R0502 41.1 twenty four 749 82900 2.61 average value 47.0 twenty four 815 89900 2.48 SEQ ID NO: 41 (196 nmol / kg) R0101 43.8 twenty four 467 43100 5.04 R0102 36.5 twenty four 645 56200 3.73 R0103 37.7 twenty four 539 45000 4.63 average value 39.3 twenty four 550 48100 4.47 SD 3.91 0.0 89.5 7080 0.670 SEQ ID NO: 42 (196 nmol / kg) R0301 33.2 48 399 38100 5.41 R0302 39.1 twenty four 559 43200 4.80 R0303 54.2 48 460 51500 4.35 average value 42.2 40 473 44300 4.85 SD 10.8 14 80.7 6760 0.532 Abbreviations: AUC0-inf = Area under the curve from 0 to infinity, CL / F = Clearance / Bioavailability, Tmax = Time to reach maximum concentration, Cmax = Maximum observed plasma concentration, T1 / 2 = Half-life. [surface]

[17] [:] [Individual and mean pharmacokinetic parameters of male Sporgodory rats after a single subcutaneous dose] [] [Compound] [(] [dose)] [Animal_ID] [T, 1 / 2 , ] [Tmax] [Cmax] [AUC, 0-inf , ] [CL / F] [(hr)] [(hr)] [(nmole / L)] [(hr*nmole / L)] [(mL / hr / kg)] SEQ ID NO: 1 (187 nmol / kg) R0101 37.7 twenty four 804 53900 3.67 R0102 38.2 twenty four 511 41200 4.77 R0103 31.2 twenty four 657 51800 3.77 average value 35.7 twenty four 657 49000 4.07 SD 3.91 0.0 147 6810 0.608 SEQ ID NO: 22 (183 nmol / kg) R0301 26.2 twenty four 783 38900 4.75 R0302 21.8 twenty four 439 34300 5.39 R0303 23.5 48 527 37100 4.99 average value 23.8 32 583 36800 5.04 SD 2.22 14 179 2320 0.323 SEQ ID NO: 25 (190 nmol / kg) R0101 14.8 twenty four 466 23400 8.13 R0102 13.9 twenty four 475 21600 8.80 R0103 14.5 twenty four 582 24700 7.69 average value 14.4 twenty four 508 23200 8.21 SD 0.458 0.0 64.5 1560 0.559 Abbreviations: AUC0-inf = area under the curve from 0 to infinity, CL / F = clearance rate / biological availability, Tmax = time to reach maximum concentration, Cmax = maximum observed plasma concentration, T1 / 2 = half-life. [surface]

[18] [:] [Individual and mean pharmacokinetic parameters of male Sporgodory rats after a single intravenous dose] [] [Compound] [(] [dose)] [Animal_ID] [T, 1 / 2 , ] [C, 0, ] [AUC, 0-inf , ] [CL] [(hr)] [(nmole / L)] [(hr*nmole / L)] [(mL / hr / kg)] SEQ ID NO: 49 (196 nmol / kg) R0001 21.7 2610 53100 3.69 R0002 21.6 3110 48400 4.05 R0003 17.8 2790 41500 4.72 average value 20.4 2840 47700 4.15 SD 2.22 253 5830 0.523 SEQ ID NO: 50 (201 nmol / kg) R0201 20.3 5560 59600 3.37 R0202 18.3 4360 74200 2.71 R0203 19.0 5960 73900 2.72 average value 19.2 5290 69200 2.93 SD 1.01 833 8340 0.378 SEQ ID NO: 29 (197 nmol / kg) R0401 41.7 6910 119000 1.66 R0402 45.2 4840 152000 1.30 R0403 49.6 1050 136000 1.45 average value 45.5 4270 136000 1.47 SD 3.96 2970 16500 0.181 SEQ ID NO: 41 (196 nmol / kg) R0001 48.4 1350 50100 3.91 R0002 49.8 1620 60600 3.23 R0003 39.0 586 45500 4.31 average value 45.7 1190 52100 3.82 SD 5.87 536 7740 0.546 SEQ ID NO: 42 (196 nmol / kg) R0201 36.1 2760 81500 2.40 R0202 35.8 2250 77900 2.51 R0204 46.0 2260 82500 2.37 average value 39.3 2420 80600 2.43 SD 5.80 292 2420 0.074 Abbreviations: AUC0-inf = Area under the curve from time 0 to infinity, CL / F = Clearance / Bioavailability, C0 = Estimated plasma concentration at time 0, T1 / 2 = Half-life. [surface]

[19] [:] [Individual and mean pharmacokinetic parameters of male Sporgodory rats after a single intravenous dose] [] [Compound] [(] [dose)] [Animal_ID] [T, 1 / 2 , ] [C, 0, ] [AUC, 0-inf , ] [CL] [(hr)] [(nmole / L)] [(hr*nmole / L)] [(mL / hr / kg)] SEQ ID NO: 1 (187 nmol / kg) R0001 23.2 6950 91900 2.04 R0003 31.0 2400 77300 2.42 R0004 47.5 5970 111000 1.69 average value 33.9 5110 93400 2.05 SD 12.4 2390 16900 0.365 SEQ ID NO: 22 (183 nmol / kg) R0201 24.7 3240 88400 2.07 R0202 21.3 5390 79600 2.30 R0203 22.9 5200 92300 1.98 average value 23.0 4610 86800 2.12 SD 1.70 1190 6510 0.165 SEQ ID NO: 25 (190 nmol / kg) R0001 17.4 1610 41300 4.59 R0002 16.9 3540 46900 4.04 R0003 18.2 3610 61800 3.07 average value 17.5 2920 50000 3.90 SD 0.656 1140 10600 0.770 Abbreviations: AUC0-inf = Area under the curve from time 0 to infinity, CL / F = Clearance / Bioavailability, C0 = Estimated plasma concentration at time 0, T1 / 2 = Half-life. [surface]

[20] [:] [Male Sporgdolly rats in a single dose]

[10] [、]

[30] [or] [100 nmol / kg] [Individual and mean pharmacokinetic parameters after subcutaneous dose] [(] [Dose Response] [)] [] [Compound] [(] [dose)] [Animal_ID] [T, 1 / 2 , ] [Tmax] [Cmax] [AUC, 0-inf , ] [CL / F] [(hr)] [(hr)] [(nmole / L)] [(hr*nmole / L)] [(mL / hr / kg)] SEQ ID NO: 1 (10 nmol / kg) R0001 26.0 twenty four 37.7 2070 5.10 R0002 41.7 twenty four 26.4 1860 6.10 R0003 31.1 twenty four 38.5 2210 4.63 average value 32.9 twenty four 34.2 2050 5.28 SD 8.01 0.0 6.77 176 0.751 SEQ ID NO: 1 (30 nmol / kg) R0101 31.5 twenty four 102 6420 4.80 R0102 37.1 twenty four 110 6920 4.55 R0103 36.1 48 104 7690 4.19 average value 34.9 32 105 7010 4.51 SD 2.99 14 4.16 640 0.307 SEQ ID NO: 1 (100 nmol / kg) R0201 30.8 twenty four 348 18800 5.46 R0202 26.0 twenty four 332 21100 4.86 R0203 24.6 twenty four 321 20700 4.91 average value 27.1 twenty four 334 20200 5.08 SD 3.25 0.0 13.6 1230 0.333 SEQ ID NO: 22 (10 nmol / kg) R0301 30.4 twenty four 49.5 2730 3.75 R0302 23.1 twenty four 51.3 2810 3.60 R0303 41.5 twenty four 48.6 3020 3.54 average value 31.7 twenty four 49.8 2850 3.63 SD 9.27 0.0 1.37 150 0.108 SEQ ID NO: 22 (30 nmol / kg) R0401 27.4 twenty four 126 7520 4.07 R0402 33.6 twenty four 132 8600 3.62 R0403 24.1 twenty four 128 7160 4.25 average value 28.4 twenty four 129 7760 3.98 SD 4.82 0.0 3.06 749 0.324 SEQ ID NO: 22 (100 nmol / kg) R0501 28.6 twenty four 372 26700 3.85 R0502 25.9 twenty four 416 26000 3.91 R0503 30.7 twenty four 369 26300 3.91 average value 28.4 twenty four 386 26300 3.89 SD 2.41 0.0 26.3 351 0.035 SEQ ID NO: 25 (10 nmol / kg) R0601 19.9 twenty four 20.8 1050 9.70 R0602 18.6 twenty four 21.9 972 10.5 R0603 31.5 6 26.8 1110 9.09 average value 23.3 18 23.2 1040 9.76 SD 7.10 10 3.19 69.2 0.707 SEQ ID NO: 25 (30 nmol / kg) R0701 19.2 twenty four 55.0 2640 11.4 R0702 19.6 twenty four 68.0 2910 10.4 R0703 20.8 6 59.1 2960 10.2 average value 19.9 18 60.7 2840 10.7 SD 0.833 10 6.65 172 0.643 SEQ ID NO: 25 (100 nmol / kg) R0801 16.0 twenty four 239 12500 7.99 R0802 17.4 twenty four 316 14000 7.15 R0803 15.7 twenty four 328 13400 7.47 average value 16.4 twenty four 294 13300 7.54 SD 0.907 0.0 48.3 755 0.424 Abbreviations: AUC0-inf = Area under the curve from 0 to infinity, CL / F = Clearance / Bioavailability, Tmax = Time to reach maximum concentration, Cmax = Maximum observed plasma concentration, T1 / 2 = Half-life. [surface] [twenty one] [:] [male] [CD-1] [Mice in a single dose] [100 nmol / kg] [Mean pharmacokinetic parameters after subcutaneous dose] [] [Compound] [(] [dose)] [T, 1 / 2 , ] [Tmax] [Cmax] [AUC, 0-inf , ] [CL / F] [(hr)] [(hr)] [(nmole / L)] [(hr*nmole / L)] [(mL / hr / kg)] SEQ ID NO: 1 (100 nmol / kg) 22.8 twenty four 327 25700 3.94 SEQ ID NO: 22 (100 nmol / kg) 25.1 twenty four 270 15000 6.76 SEQ ID NO: 25 (100 nmol / kg) 32.5 twenty four 366 25000 4.01 Abbreviations: AUC0-inf = Area under the curve from 0 to infinity, CL / F = Scavenging rate / Bioavailability, Tmax = Time to reach maximum concentration, Cmax = Maximum observed concentration, T1 / 2 = Half-life. PK Conclusion

[0293] In different species, compared with wild-type UCN2, the pharmacokinetic profiles showed a prolonged elimination half-life and reduced clearance. These data indicate that the above compounds have a pharmacokinetic profile suitable for once-weekly administration. [Example] [8] [:] [Immunogenicity Risk Assessment] MAPPS analysis (MHC-related peptide proteomics):

[0294] MAPPS characterizes MHC-II-presented peptides on human dendritic cells treated with the test molecule. CD14+ cells were isolated from normal human donor PBMCs, cultured, and differentiated into immature dendritic cells (DCs) by co-culturing with IL-4 and GM-CSF. On day 4, the medium was replaced with fresh medium containing the test molecule. On day 5, LPS was added to convert the cells into mature DCs. On day 6, cells were lysed with RIPA buffer containing a protease inhibitor. Immunoprecipitation of the MHC-II complex was performed using a biotinylated anti-MHC-II antibody conjugated to streptobiotin beads. The bound complex was dissociated and filtered. The isolated MHC-II peptides were analyzed by mass spectrometry. Peptide identification was generated using an enzyme-free search algorithm and a bovine / human database with accompanying test sequences via an internal proteomics workflow. The identification profile of the samples was processed using the KNIME workflow. The peptides identified by the self-test were compared with the parental sequences of the test molecule. The output results were used to determine the percentage of donor peptides displaying MHC-II peptides from the regions of the test molecule. Peptides containing SEQ ID NO: 54, 49, 50, 29, 41, 45, 1, 22, and 25 were tested in MAPP analysis. Peptides containing SEQ ID NO: 41, 45, and 25 did not show up on the MHC-II complex in the analysis. Compounds containing SEQ ID NO: 54, 49, 50, 29, 1, and 22 displayed at least one peptide on the MHC-II complex of the donor subgroup. Depending on the donor, up to three major clusters were observed, with slight offsets between them, spanning a total of 1 to 33 residues. T cell proliferation analysis

[0295] This analysis evaluates the ability of the test antibody or the test MAPPS peptide to activate CD4+ T cells by inducing cell proliferation. PBMCs depleted of CD8+ T cells and labeled with CFSE were prepared. Each sample was tested using a culture medium control, keyhole hemocyanin (KLH; positive clinical baseline control), the test antibody, or the test MAPPS peptide. Cultures were incubated for 7 days. On day 7, samples were analyzed by flow cytometry. Reference analogs containing SEQ ID NO: 54 elicited a reaction in four out of ten donors, while analogs of the present invention did not elicit a reaction. Pre-existing reactivity (ACE analysis format)

[0296] This analysis assesses pre-existing antibodies (ADAs) against the test molecule in untreated, healthy human serum (NHS). Diluted NHS was incubated overnight on Pierce streptavidin disks coated with biotinylated test molecules. The next day, the captured binding protein was acid-dissociated, firmly coated onto Mesoscale (MSD) disks, and detected using a combination of the biotinylated molecule and ruthenium-labeled streptavidin. If the anti-drug antibody is present, it will bind to the labeled drug, and the resulting signal is termed a Grade 1 signal (expressed as electrochemiluminescence). In Grade 2, the Grade 1 signal is confirmed by the addition of an excess of unlabeled test molecule during the detection step, which inhibits the Grade 1 signal. The presence of pre-existing anti-drug antibodies is expressed as the 90th percentile of Grade 2 inhibition. The 90th percentile of Grade 2 inhibition is a statistical tool for assessing the specificity of Grade 1 reactivity. This 90th percentile was used to rank molecules according to ADA risk. The 90th percentile of second-order inhibition of the peptide containing SEQ ID NO: 1 is higher than that of the peptides containing SEQ ID NO: 22 and 25. [surface] [twenty two.] [Overview of Immunogenicity Risk Assessment] [SEQ ID NO] [MAPPS] [analyze] [T] [Cell proliferation analysis] [:] [protein] [Pre-existing reactivity] [] >10 donors with different MHC 7 to 10 donors with different MHCs 90th percentile T2 inhibition > 50 donors 54 7 / 10 (70%) 4 / 10 ND 49 6 / 26 (23%) ND ND 50 3 / 10 (30%) ND ND 29 8 / 30 (27%) ND ND 41 0 / 10 (0%) ND ND 45 0 / 20 (0%) ND ND 1 3 / 20 (15%) 0 / 9 37.9% twenty two 1 / 10 (10%) 0 / 9 14.3% 25 0 / 10 (0%) 0 / 9 4.5% Abbreviations: ACE = Acid-Captured Dissociation; ADA = Anti-Drug Antibody; CDR = Complementarity-Determining Region; DC = Dendritic Cell; H1 = VH CDR1; H2 = VH CDR2; H3 = VH CDR3; L1 = VL CDR1; L2 = VL CDR2; MAPR = MHC-Related Peptide Proteomics; MHC = Major Histocompatibility Complex; MS = Mass Spectrometry; ND = No Data; T2 = Grade 2; TCEM = T Cell Exposure Phantom; VH = Variable Heavy Chain; VH = Variable Light Chain; VHFR3 = Variable Heavy Chain Framework 3 In MAPPs analysis and / or T cell proliferation analysis, peptides containing SEQ ID NO: 49, 50, 29, 41, 45, 1, 22 and 25 showed a substantial improvement in immunogenicity compared to the compound containing SEQ ID NO: 54. [sequence] [] [SEQ ID NO: 1] Ac-GSPSIVTS αMeL DVPT Aib αMeL LQKILEQERAEKARQQAK(γE-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-εK-CO-(CH2)18-CO2H)EAAEILAQV-NH2 [SEQ ID NO: 2] Ac-GSPSIVTS αMeL DVPT Aib αMeL LQKLLEQERAEKARQQAK(γE-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-εK-CO-(CH2)18-CO2H)EAAEILAQV-NH2 [SEQ ID NO: 3] Ac-GSPSIVTS αMeL DVPT Aib αMeL LQKILEQERAEKARQQAK((2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-PKE-γE-CO-(CH2)18-CO2H)EAAEILAQV-NH2 [SEQ ID NO: 4] Ac-GSPSIVTS αMeL DVPT Aib αMeL LQKILEQERAEKARQQAK(Ahx-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-γE-CO-(CH2)18-CO2H)EAAEILAQV-NH2 [SEQ ID NO: 5] Ac-GSPSIVTS αMeL DVPT Aib αMeL LQKILEQERAEKARQQAK(γE-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-Ahx-CO-(CH2)18-CO2H)EAAEILAQV-NH2 [SEQ ID NO: 6] Ac-GSPSIVTS αMeL DVPT Aib αMeL LQKILEQERAEKARQQAK(γE-GKEKEKE-CO-(CH2)18-CO2H)EAAEILAQV-NH2 [SEQ ID NO: 7] Ac-GSPSIVTS αMeL DVPT Aib αMeL LQKILEQERAEKARQQAK(G-(βA)3-γE-CO-(CH2)18-CO2H)EAAEILAQV-NH2 [SEQ ID NO: 8] Ac-GSPSIVTS αMeL DVPT Aib αMeL LQKILEQERAEKARQQAK(G-(βA)4-γE-CO-(CH2)18-CO2H)EAAEILAQV-NH2 [SEQ ID NO: 9] Ac-GSPSIVTS αMeL DVPT Aib αMeL LQKILEQERAEKARQQAK((2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-KEKEKE-γE-CO-(CH2)18-CO2H)EAAEILAQV-NH2 [SEQ ID NO: 10] Ac-GSPSIVTS αMeL DVPT Aib αMeL LQKILEQERAEKARQQAK((βA)-S-(βA)-S-γE-CO-(CH2)18-CO2H)EAAEILAQV-NH2 [SEQ ID NO: 11] Ac-GSPSIVTS αMeL DVPT Aib αMeL LQKILEQERAEKARQQAK(γE-PKE-Ahx-CO-(CH2)18-CO2H)EAAEILAQV-NH2 [SEQ ID NO: 12] Ac-GSPSIVTS αMeL DVPT Aib αMeL LQKLLEQERAEKARQQAK(γE-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-εK-CO-(CH2)16-CO2H)EAAEILAQV-NH2 [SEQ ID NO: 13] Ac-GSPSIVTS αMeL DVPT Aib αMeL LQKILEQERAEKARQQAK(γE-PKE-εK-CO-(CH2)18-CO2H)EAAEILAQV-NH2 [SEQ ID NO: 14] Ac-GSPSIVTS αMeL DVPT Aib αMeL LQKILEQERAEKARQQAK(γE-PPP-εK-CO-(CH2)18-CO2H)EAAEILAQV-NH2 [SEQ ID NO: 15] Ac-GSPSIVTS αMeL DVPT Aib αMeL LQKILEQERAEKARQQAK(γE-PPPPPP-CO-(CH2)18-CO2H)EAAEILAQV-NH2 [SEQ ID NO: 16] Ac-GSPSIVTS αMeL DVPT Aib αMeL LQKILEQERAEKARQQAK(γE-PEPEPE-CO-(CH2)18-CO2H)EAAEILAQV-NH2 [SEQ ID NO: 17] Ac-GSPSIVTS αMeL DVPT Aib αMeL LQKILEQERAEKARQQAK(γE-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)2-εK-CO-(CH2)18-CO2H)EAAEILAQV-NH2 [SEQ ID NO: 18] Ac-GSPSIVTS αMeL DVPT Aib αMeL LQKILEQERAEKARQQAK(γE-εK-yE-εK-CO-(CH2)18-CO2H)EAAEILAQV-NH2 [SEQ ID NO: 19] Ac-GSPSIVTS αMeL DVPT Aib αMeL LQKILEQERAEKARQQAK(γE-εK-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-εK-CO-(CH2)18-CO2H)EAAEILAQV-NH2 [SEQ ID NO: 20] Ac-GSPSIVTS αMeL DVPT Aib αMeL LQKILEQERAEKARQQAKEAAEILAQV-NH2 [SEQ ID NO: 21] Ac-GSPSIVTS αMeL DVPT Aib αMeL LQKILEQERAEKARQQAK(γE- (2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-εK-CO-(CH2)16-CO2H)EAAEILAQV-NH2 [SEQ ID NO: 22] Pyr-GSPSGIVTS αMeL DVPT Aib αMeL LQKILEQERAEKARQQAK(γE-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-εK-CO-(CH2)18-CO2H)EAAEILAQV-NH2 [SEQ ID NO: 23] Pyr-GSPSGIVTS αMeL DVPT Aib αMeL LQKILEQERAEKARQQAKEAAEILAQV-NH2 [SEQ ID NO: 24] Pyr-GSPSGIVTS αMeL DVPT Aib αMeL LQKILEQERAEKARQQAK(γE-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-εK-CO-(CH2)16-CO2H)EAAEILAQV-NH2 [SEQ ID NO: 25] HO2C-(CH2)18-CO-εK-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-γE-GSPSIVTS αMeL DVPT Aib αMeL LQKILEQERAEKARQQATEAAEILAQV-NH2 [SEQ ID NO: 26] HO2C-(CH2)18-CO-K-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-γE-GSPSIVTS αMeL DVPT Aib αMeL LQKILEQERAEKARQQATEAAEILAQV-NH2 [SEQ ID NO: 27] GSPSIVTS αMeL DVPT Aib αMeL LQKILEQERAEKARQQATEAAEILAQV-NH2 [SEQ ID NO: 28] HO2C-(CH2)16-CO-εK-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-γE-GSPSIVTS αMeL DVPT Aib αMeL LQKILEQERAEKARQQATEAAEILAQV-NH2 [SEQ ID NO: 29] Ac-IVTS αMeL D (D-Val) PTG αMeL LQIKLEQERAEKARQ QAK((2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)2-γE-CO-(CH2)18-CO2H)ENTEILAQV-NH2 [SEQ ID NO: 30] Ac-IVTS αMeL D (D-Val) PTG αMeL LQIKLEQERAEKARQQAK(γE-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-γE-CO-(CH2)18-CO2H)ENTEILAQV-NH2 [SEQ ID NO: 31] Ac-IVTS αMeL D (D-Val) PTG αMeL LQIKLEQERAEKARQQAK((γE)2-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-CO-(CH2)18-CO2H)ENTEILAQV-NH2 [SEQ ID NO: 32] Ac-IVTS αMeL D (D-Val) PTG αMeL LQIKLEQERAEKARQQAK((2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-εK-γE-CO-(CH2)18-CO2H)ENTEILAQV-NH2 [SEQ ID NO: 33] Ac-IVTS αMeL D (D-Val) PTG αMeL LQIKLEQERAEKARQQAK((2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-γE-εK-CO-(CH2)18-CO2H)ENTEILAQV-NH2 [SEQ ID NO: 34] Ac-IVTS αMeL D (D-Val) PTG αMeL LQIKLEQERAEKARQQAK(εK-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-γE-CO-(CH2)18-CO2H)ENTEILAQV-NH2 [SEQ ID NO: 35] Ac-IVTS αMeL D (D-Val) PTG αMeL LQIKLEQERAEKARQQAK(εK-γE-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-CO-(CH2)18-CO2H)ENTEILAQV-NH2 [SEQ ID NO: 36] Ac-IVTS αMeL D (D-Val) PTG αMeL LQIKLEQERAEKARQQAK(γE-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-εK-CO-(CH2)18-CO2H)ENTEILAQV-NH2 [SEQ ID NO: 37] Ac-IVTS αMeL D (D-Val) PTG αMeL LQIKLEQERAEKARQQAK(γE-εK-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-CO-(CH2)18-CO2H)ENTEILAQV-NH2 [SEQ ID NO: 38] Ac-IVTS αMeL D (D-Val) PTG αMeL LQIKLEQERAEKARQQAK(γE-γE-εK-CO-(CH2)18-CO2H)ENTEILAQV-NH2 [SEQ ID NO: 39] Ac-IVTS αMeL D (D-Val) PTG αMeL LQIKLEQERAEKARQQAK(γE-εK-γE-CO-(CH2)18-CO2H)ENTEILAQV-NH2 [SEQ ID NO: 40] Ac-IVTS αMeL D (D-Val) PTG αMeL LQIKLEQERAEKARQQAK(εK-(γE)2-CO-(CH2)18-CO2H)ENTEILAQV-NH2 [SEQ ID NO: 41] [[ID=​​​​​​​​​​​​​​​​​​​γE-GPSIVTS αMeL D (D-Val) PI Aib αMeL LQIKLEQERAEKARQQAK((2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)2-γE-CO-(CH2)18-CO2H)ENTEILAQV-NH2 [SEQ ID NO: 46] Ac-PPPIVTS αMeL D (D-Val) PT Aib αMeL LQK Aib LEQERAEKARQQAK(γE-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-εK-CO-(CH2)18-CO2H)ENTEILAQV-NH2 [SEQ ID NO: 47] Ac-PPPIVTS αMeL D (D-Val) PT Aib αMeL LQKLLEQERAEKARQQAK(γE-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-εK-CO-(CH2)18-CO2H)EAAEILAQV-NH2 [SEQ ID NO: 48] Ac-PPPIVTS αMeL DVPT Aib αMeL LQKLLEQERAEKARQQAK(γE-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-εK-CO-(CH2)18-CO2H)EAAEILAQV-NH2 [SEQ ID NO: 49] (NMeI) VTSLDVPIG αMeL LQKLIEQEKQEKEKQQAK((2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)2-γE-CO-(CH2)18-CO2H)ENTEILAQV-NH2 [SEQ ID NO: 50] (NMeI) VTSLDVPIG αMeL LQKLLEQERAEKARQQAK((2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)2-γE-CO-(CH2)18-CO2H)EAAEILAQV-NH2 [SEQ ID NO: 51] Ac-IVTS αMeL D (D-Val) PI Aib αMeL LQIKLEQERAEKARQQAK((γE)2-CO-(CH2)18-CO2H)ENTEILAQV-NH2 [SEQ ID NO: 52] HO2C-(CH2)18-CO-Dap-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-γE-GSPSIVTS αMeL DVPT Aib αMeL LEKILEQERAEKARQQATEAAEILAQV-NH2 [SEQ ID NO: 53] IVLSLDVPIGLLQILLEQARARAAREQATTNARILARV-NH2 [SEQ ID NO: 54] (NMeI) VLSLDVPIGLLQILLEQEKQEKEKQQAK((2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)2-(γE)2-CO-(CH2)18-CO2H)TNAQILAQV-NH2 [SEQ ID NO: 55] Y Aib EGTFTSDYSI Aib LDKIAQK((2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)2-γE-CO-(CH2)18-CO2H)AFVQWLIAGGPSSGAPPPS-NH2 [SEQ ID NO: 56] Y Aib QGTFTSDYSI αMeL LDKK((2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)2-γE-CO-(CH2)18-CO2H)AQ Aib AFIEYLLEGGPSSGAPPPS-NH2 [SEQ ID NO: 57] Y Aib QGTFTSDYSI αMeL LDKK((2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-γE-CO-(CH2)18-CO2H)AQ Aib AFIEYLLEGGPSSGAPPPS-NH2 [SEQ ID NO: 58] Y Aib QGTFTSDYSILLDKK((2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)2-γE-CO-(CH2)18-CO2H)AQ Aib AFIEYLLEGGPSSGAPPPS-NH2 [SEQ ID NO: 59] H Aib QGTFTSDYSKYLDEKKAK((2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)2-γE-CO-(CH2)16-CO2H)EFVEWLLEGGPSSG-NH2 [SEQ ID NO: 60] H Aib QGTFTSDYSKYLDEKKAK((2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)2-(γE)2-CO-(CH2)16-CO2H)EFVEWLLSGGPSSG-NH2 [SEQ ID NO: 61] H Aib QGTFTSDYSKYLDEKKAK((2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)2-γE-CO-(CH2)18-CO2H)EFVEWLLEGGPSSG-NH2 [SEQ ID NO: 62] H Aib QGTFTSDYSKYLDEKKAK((2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)2-(γE)2-CO-(CH2)18-CO2H)EFVEWLLSGGPSSG-NH2 [SEQ ID NO: 63] γE-[CNTATC]*ATG Orn LAE αMeF LVRSSN (NMeN) FGPK((γE)2-CO-(CH2)18-CO2H)LPPTEVGSNTY-NH2 [SEQ ID NO: 64] γE-[CNTATC]*ATG Orn LAE αMeF LVRSSN (NMeD) FGPK((γE)2-CO-(CH2)18-CO2H)LPPTEVGSNTY-NH2 [SEQ ID NO: 65] K((γE)2-CO-(CH2)18-CO2H)-[CETATC]*ATG Orn LAE αMeF LVRSSN (NMeD) FGPILPPTEVGSNTY-NH2 [SEQ ID NO: 66] K((γE)2-CO-(CH2)18-CO2H)-[CETATC]*ATG Orn αMeL AEFLVRSSHNFGPILPPTEVGSNTY-NH2 [SEQ ID NO: 67] Ac-ASHLSTAVLGK((2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)2-γE-CO-(CH2)18-CO2H) LS Aib ELHKLEDYPRTDVGAESP-NH2 [SEQ ID NO: 68] X1X2X3X4X5X6IVTSX11DX13PX15X16X17LX19X20X21X22EQEX26X27EKX30X31QQAX35EX37X38EILAQV, where X1 is Pyr, E, γE, or does not exist. X2 is G or does not exist. X3 is G, S, or does not exist. X4 is S, P, G, or does not exist. X5 is S, P, or does not exist. X6 is G, S, P, or does not exist. X11 is L or αMeL, X13 is either V or D-Val. X15 is either T or I. X16 is either Aib or G. X17 is either L or αMeL. X19 is either Q or E. X20 is either K or I. X21 can be I, K, Aib, or L. X22 represents I or L. X26 is either R or K. X27 is either A or Q. X30 is either A or E. X31 is either R or K. X35 is either K or T. X37 is either A or N. X38 is either A or T. The N-terminal amino acid is N-acylated, N-acetylated, or N-methylated as appropriate, and the C-terminal amino acid is amylated as appropriate. [SEQ ID NO: 69] GSPSIVTS αMeL DVPT Aib αMeL LQKX21LEQERAEKARQQAKEAAEILAQV, where X21 is either I or L. The N-terminal amino acid is N-acetylated as appropriate, and the C-terminal amino acid is amylated as appropriate. [] [SEQ ID NO: 70] Pyr GSPSGIVTS αMeL DVPT Aib αMeL LQKILEQERAEKARQQAKEAAEILAQV, The C-terminal amino acid is acetaminated as appropriate. [SEQ ID NO: 71] X1X2GSPSIVTS αMeL DVPT Aib αMeLLX19KILEQERAEKARQQATEAAEILAQV, where X1 is either γE or does not exist. X2 does not exist. X19 is either Q or E. The C-terminal amino acid is acetaminated as appropriate. [SEQ ID NO: 72] IVTS αMeL D D-Val PX15X16αMeLLQIKLEQERAEKARQQAKENTEILAQV (SEQ ID NO: 72), where X15 is either T or I. X16 is either G or Aib. The N-terminal amino acid is N-acetylated as appropriate, and the C-terminal amino acid is amylated as appropriate. [SEQ ID NO: 73, <s> < / s> , ] PPPIVTS αMeL DX13PTAib αMeL LQKX21LEQERAEKARQQAKEX37X38EILAQV, where X13 is either V or D-Val. X21 is either L or Aib. X37 is either A or N. X38 is either A or T. The N-terminal amino acid is N-acetylated as appropriate, and the C-terminal amino acid is amylated as appropriate. [SEQ ID NO: 74] γE-GKEKEKE-CO-(CH2)18-CO2H [SEQ ID NO: 75] (2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-KEKEKE-γE-CO-(CH2)18-CO2H [SEQ ID NO: 76] γE-PPPPPP-CO-(CH2)18-CO2H [SEQ ID NO: 77] γE-PEPEPE-CO-(CH2)18-CO2H [SEQ ID NO: 78] GKEKEKE-γE-CO-(CH2)18-CO2H [SEQ ID NO: 79] H Aib EGTFTSDVSSYLEGQAAK((2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)2-γE-CO-(CH2)16-CO2H)EFIAWLVRGRG-OH [SEQ ID NO: 80] HAEGTFTSDVSSYLEGQAAK(γE-CO-(CH2)14-CH3)EFIAWLVRGRG-OH [SEQ ID NO: 81] Dulaglutide is a human GLP-1 receptor agonist comprising a dimer of a GLP-L analog fused at its C-terminus to the N-terminus of an immunoglobulin Fc fragment analog via a peptide linker. Dulaglutide is identified by CAS registry number 923950-08-7 and is provided with the following chemical name: 7-37-I [8-glycine, 22-glutamic acid, 36-glycine] (synthetic human) fusion protein and peptide (synthetic 16-amino acid linker) fusion protein and immunoglobulin G4 (synthetic human Fc fragment), dimer. Each monomer of dulaglutide has the amino acid sequence described in SEQ ID NO: 81. HGEGTFTSDVSSYLEEQAAKEFIAWLVKGGGGGGGSGGGGSGGGGSAESKYGPPCPPCPAPEAAGGPSVFLFPPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTV LHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG

[0297] TW202540149A_113143996_SEQL.xml

Claims

1. A polypeptide or a pharmaceutically acceptable salt thereof active against corticotropin-releasing hormone receptor-2 (CRHR2), comprising: X1X2X3X4X5X6IVTSX11DX13PX15X16X17LX19X20X21X22EQEX26X27EKX30X31QQAX35EX37X38EILAQV (SEQ ID) NO: 68), where a) X1 does not exist, X2 does not exist, X3 is G, X4 is S, X5 is P, X6 is S, X11 is αMeL, X13 is V, X15 is T, X16 is Aib, X17 is αMeL, X19 is Q, X20 is K, X21 is I or L, X22 is L, X26 is R, X27 is A, X30 is A, X31 is R, X35 is K, X37 is A, and X38 is A; or b) X1 is Pyr, X2 is G, X3 is S, X4 is P, X5 is S, X6 is G, X11 is αMeL, X13 is V, X15 is T, X16 is Aib, X17 is αMeL, X19 is Q, X20 is K, X21 is I, X22 is L, X26 is R, X27 is A, X30 is A, X31 is R, X35 is K, X37 is A, and X38 is A; or c) X1 is γE or does not exist, X2 does not exist, X3 is G, X4 is S, X5 is P, X6 is S, X11 is αMeL, X13 is V, X15 is T, X16 is Aib, X17 is αMeL, X19 is Q or E, X20 X21 is K, X22 is I, X26 is R, X27 is A, X30 is A, X31 is R, X35 is T, X37 is A, and X38 is A; or d) X1 does not exist, X2 does not exist, X3 does not exist, X4 does not exist, X5 does not exist, X6 does not exist, X11 is αMeL, X13 is D-Val, X15 is T or I, X16 is Aib or G, X19 is Q, X20 is I, X21 is K, X22 is L, X26 is R, X27 is A, X30 is A, X31 is R, X35 is K, X 37 is N, and X38 is T; or e) X1 does not exist, X2 does not exist, X3 does not exist, X4 is P, X5 is P, X6 is P, X11 is αMeL, X13 is V or D-Val, X15 is T, X16 is Aib, X17 is αMeL, X19 is Q, X20 is K, X21 is L or Aib, X22 is L, X26 is R, X27 is A, X30 is A, X31 is R, X35 is K, X37 is A or N, X38 is A or T; or f) X1 is γE, X2 does not exist, X3 does not exist;X4 is G, X5 is P, X6 is S, X11 is αMeL, X13 is D-Val, X15 is I, X16 is Aib, X17 is αMeL, X19 is Q, X20 is I, X21 is K, X22 is L, X26 is R, X27 is A, X30 is A, X31 is R, X35 is K, X37 is N, X38 is T; or g) X1 does not exist, X2 does not exist, X3 does not exist, X4 does not exist, X5 does not exist, X6 does not exist, X11 is L, X13 is V, X15 is I, X16 is G, X17 is αMeL, X19 is Q, X20 is K, X21 is L, X22 is I, X26 is K, X27 is Q, X30 is E, X31 is K, X35 is K, X37 is N, X38 is T; or h) X1 does not exist, X2 does not exist, X3 does not exist, X4 does not exist, X5 does not exist, X6 does not exist, X11 is L, X13 is V, X15 is I, X16 is G, X17 is αMeL, X19 is Q, X20 is K, X21 is L, X22 is L, X26 is R, X27 is A, X30 is A, X31 is R, X35 is K, X37 is A, X38 is A.

2. The polypeptide or its pharmaceutically acceptable salt as claimed in claim 1, wherein X1 is absent, X2 is absent, X3 is G, X4 is S, X5 is P, X6 is S, X11 is αMeL, X13 is V, X15 is T, X16 is Aib, X17 is αMeL, X19 is Q, X20 is K, X21 is I or L, X22 is L, X26 is R, X27 is A, X30 is A, X31 is R, X35 is K, X37 is A, and X38 is A.

3. The polypeptide of claim 2 or a pharmaceutically acceptable salt thereof, wherein X21 is I.

4. The polypeptide or its pharmaceutically acceptable salt as claimed in claim 2, wherein X21 is L.

5. The polypeptide of claim 1 or a pharmaceutically acceptable salt thereof, wherein X1 is γE or absent, X2 is absent, X3 is G, X4 is S, X5 is P, X6 is S, X11 is αMeL, X13 is V, X15 is T, X16 is Aib, X17 is αMeL, X19 is Q or E, X20 is K, X21 is I, X22 is L, X26 is R, X27 is A, X30 is A, X31 is R, X35 is T, X37 is A, and X38 is A.

6. The polypeptide or its pharmaceutically acceptable salt as claimed in claim 5, wherein X19 is Q.

7. The polypeptide of claim 2 or a pharmaceutically acceptable salt thereof, wherein X19 is E.

8. The polypeptide of claim 1 or a pharmaceutically acceptable salt thereof, further comprising a fatty acid, wherein the fatty acid is bound to the polypeptide via a direct bond or linker between the amino acid and the fatty acid at an amino acid having a functional group available for binding.

9. The polypeptide of claim 1 or a pharmaceutically acceptable salt thereof, wherein the N-terminal amino acid is N-acetylated.

10. The polypeptide of claim 1 or a pharmaceutically acceptable salt thereof, wherein the N-terminal amino acid is N-methylated.

11. The polypeptide of claim 1 or a pharmaceutically acceptable salt thereof, wherein the C-terminal amino acid is acetaminated.

12. A polypeptide active against corticotropin-releasing hormone receptor-2 (CRHR2) or a pharmaceutically acceptable salt thereof, wherein the polypeptide comprises any one of SEQ ID NO: 1 to 52.

13. The polypeptide of claim 12 or a pharmaceutically acceptable salt thereof, wherein the polypeptide is selected from the group consisting of: SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 22, SEQ ID NO: 25, SEQ ID NO: 29, SEQ ID NO: 41, SEQ ID NO: 42, SEQ ID NO: 45, SEQ ID NO: 46, SEQ ID NO: 48, SEQ ID NO: 49 and SEQ ID NO:

50.

14. A pharmaceutical composition comprising a polypeptide or a pharmaceutically acceptable salt thereof as claimed in any one of claims 1 to 13 and at least one pharmaceutically acceptable carrier, diluent or excipient.

15. Use of a polypeptide or a pharmaceutically acceptable salt thereof as claimed in any one of claims 1 to 13 for the manufacture of an agent for treating diseases or conditions selected from the group consisting of: diabetes, obesity, chronic weight management, non-alcoholic fatty liver disease (NAFLD), non-alcoholic steatohepatitis (NASH), dyslipidemia, metabolic syndrome, chronic kidney disease (CKD), osteoarthritis (OA), obesity-related sleep apnea (OSA), sarcopenia, cachexia, sarcopenic obesity (SO), and polycystic ovarian syndrome (PCOS).

16. As requested in claim 15, wherein the agent is used in combination with an additional therapeutic agent.

17. As claimed in claim 16, wherein the additional therapeutic agent is a dual agonist of the GIP and GLP-1 receptors.

18. As claimed in claim 17, wherein the dual agonist of the GIP and GLP-1 receptor is a compound comprising SEQ ID NO:

55.

19. As claimed in claim 16, wherein the additional therapeutic agent is a GLP-1 receptor agonist.

20. The use as claimed in claim 19, wherein the GLP-1 receptor agonist is a compound comprising SEQ ID NO: 79, SEQ ID NO: 80 or SEQ ID NO: 81.