Compounds for the treatment of diabetes or obesity

Abstract

To provide novel urocortin-2 polypeptides having activity at the corticotropin-releasing hormone receptor-2 (CRHR2), pharmaceutical compositions, and methods of using the same.SOLUTION: A polypeptide, or a pharmaceutically acceptable salt thereof comprises: X1X2X3X4X5X6IVTSX11DX13 PX15X16X17LX19X20X21X22EQEX26X27EKX30X31QQAX35EX37X38EILAQV, where X1 is Pyr, E, γE, or absent, X2 is G or absent, X3 is G, S, or absent, X4 is S, P, G, or absent, X5 is S, P, or absent, X6 is G, S, P, or absent, the N-terminal amino acid is optionally N-acylated, N-acetylated or N-methylated, and the C-terminal amino acid is optionally amidated.SELECTED DRAWING: None

Description

[Technical Field]

[0001] This application is filed together with a sequence list in ST.26 XML format. The sequence list is prepared as a file titled "30698 WO SL" created on October 31, 2024, and is 2.48 megabytes in size. The ST.26 XML format sequence list information is incorporated herein by reference in its entirety.

[0002] This disclosure relates to a polypeptide having activity at the corticotropin-releasing hormone receptor-2 (CRHR2), a pharmaceutical composition containing the polypeptide, and a method of using the polypeptide to treat a disorder related thereto. [Background technology]

[0003] Diabetes, obesity, and chronic kidney disease (CKD) are widespread diseases worldwide that present significant associated health challenges. While they are distinct conditions, they often occur together as comorbidities.

[0004] Diabetes is a metabolic disorder that occurs when the pancreas does not produce enough insulin, or when the body needs more insulin than it can produce, leading to hyperglycemia or high blood glucose. In 2021, the U.S. Centers for Disease Control and Prevention (CDC) estimated that approximately 29.7 million people in the U.S., or 8.9% of the population, were diagnosed with diabetes. Even more worrying, it is estimated that approximately 38.0% of adults aged 18 and over in the U.S. are prediabetic. One of the main risk factors for diabetes is obesity.

[0005] Obesity is a medical disorder resulting from the excessive accumulation of adipose tissue. Today, obesity is a global public health concern, which the CDC estimates affects approximately 40% of the adult population. Furthermore, obesity is associated with high costs and serious undesirable health outcomes and pathological conditions. Some obese individuals can be further characterized as having a subset of obesity, such as sarcopenic obesity, which is a condition characterized by 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, for all patients with obesity, high-quality weight loss, i.e., loss of adipose tissue with little to no impact on lean body mass, is crucial.

[0006] Furthermore, both diabetes and obesity are risk factors for chronic kidney disease (CKD), a condition characterized by progressive loss of kidney function. Over time, individuals with CKD experience increased albuminuria, proteinuria, serum creatinine, and / or renal histopathological lesions. For many patients, CKD eventually progresses to end-stage renal disease (ESRD), requiring either dialysis or a kidney transplant.

[0007] The challenges associated with managing diabetes, obesity, and / or chronic kidney disease include the need to address one or more underlying factors contributing to each patient's specific disease (e.g., high blood sugar, overweight, and excess body fat relative to lean body mass).

[0008] Currently, patients with diabetes, obesity, and / or chronic kidney disease have numerous treatment options, including incretin agonists (e.g., liraglutide, tilzepatide, semaglutide), amylin agonists (e.g., pramulintide), DPP-4 inhibitors, metformin, insulin, and many others. However, currently available treatments may present numerous challenges in some patients, including short half-lives, the need for frequent dosing, unpleasant side effects such as gastrointestinal disturbances, vomiting, and / or hypoglycemic reactions, immunogenic reactions, and / or the inability to effectively treat one or more cofactors related to the condition (e.g., hyperglycemia, overweight, and the presence of excess fat relative to lean body mass).

[0009] 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 others being UCN1 and UCN3) and is part of the corticotropin-releasing hormone (CRH) family. Urocortin peptides are short-acting peptides that act through CRH receptors known as CRHR1 and / or CRHR2. UCN2 has been reported to improve insulin sensitivity and glucose tolerance through its action as a ligand for the G protein-coupled receptor corticotropin-releasing hormone receptor 2 (CRHR2), and therefore polypeptides active at the CRHR2 receptor are being sought as therapeutic agents for diabetes and related diseases. Chen et al. 16580-16585 (2006). However, although urocortin analogs are known in the art (see, for example, U.S. Patent No. 10,894,817 (Alsina-Fernandez, Eli Lilly), International Publication No. 2023 / 285334 (Li, Novo Nordisk), and International Publication No. 2023 / 161229 (Janiak, Corteria Pharmaceuticals)), urocortin analogs are not currently approved for medical use.

[0010] With this in mind, polypeptides that are active at the CRH2 receptor are needed. Specifically, therapeutic compounds are needed that can treat obesity, diabetes, and / or CKD while maintaining lean body mass. Furthermore, compounds are needed that possess one or more of the following characteristics: a favorable in vitro immunogenicity profile, selective efficacy against the CRH2 receptor, improved PK (half-life) compared to endogenous human urocortin, high-quality weight loss with maintenance of lean body mass compared to fat mass, and favorable developmental characteristics (e.g., association state). [Overview of the Initiative]

[0011] The present specification provides a polypeptide having activity at the CRH2 receptor for the treatment of diabetes, obesity, and / or CKD, which also addresses one or more of the above problems. According to an embodiment of the present disclosure, provided herein is a polypeptide of Formula I, or a pharmaceutically acceptable salt thereof, X1X2X3X4X5X6IVTSX 11 DX 13 PX 15 X 16 X 17 LX 19 X 20 X 21 X 22 EQEX 26 X 27 EKX 30 X 31 QQAX 35 EX 37 X 38 EILAQV (SEQ ID NO: 68), X1 is Pyr, E, γE, or is absent, X2 is G, or is absent, X3 is G, S, or is absent, X4 is S, P, G, or is absent, X5 is S, P, or is absent, X6 is G, S, P, or is absent, X 11 is L or αMeL, X 13 is V or D-Val, X 15 is T or I, X 16 is Aib or G, X 17 is L or αMeL, X 19 is Q or E, X 20 is K or I, X 21 is I, K, Aib, or L, X 22 is I or L, X26 However, it is R or K, X 27 However, it is either A or Q, X 30 However, it is A or E, X 31 However, it is R or K, X 35 However, it is either K or T, X 37 However, it is either A or N, X 38 However, it is A or T, Polypeptides or pharmaceutically acceptable salts thereof are provided, wherein the N-terminal amino acid is optionally N-acylated, N-acetylated, or N-methylated, and the C-terminal amino acid is optionally amidated.

[0012] In some embodiments of Formula I, the V at position 44 is the C-terminal amino acid. The C-terminus may contain an amide, carboxylic acid, or other modification. The N-terminal amino acid is embodiment-dependent but is always the first amino acid present in the sequence that indicates 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; acylation, or other modifications). Furthermore, if any amino acid at positions X1-X6 is indicated as "absent," the next available amino acid conjugates with the previously available amino acid. Thus, various embodiments of Formula I result in polypeptides of varying lengths, e.g., 38-44 amino acids.

[0013] Furthermore, in addition to the sequences described herein, the polypeptides described herein may contain one or more conservative amino acid substitutions, provided that the polypeptide remains capable of being active in CRHR2.

[0014] In some embodiments, the polypeptide, or a pharmaceutically acceptable salt thereof, further includes means for increasing the half-life of the polypeptide. In some embodiments, the polypeptide, or a pharmaceutically acceptable salt thereof, further includes means for increasing the half-life of the polypeptide via any amino acid on the polypeptide having a functional group suitable for conjugation. In some embodiments, the amino acid having a functional group suitable for conjugation is γE, E, or K. In some embodiments, the functional group available for conjugation conjugates with phosphonates, tetrazoles, 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, providing the potential to further generate long-acting compounds.

[0015] In some embodiments, these means are fatty acids.

[0016] In some embodiments, the amino acid having a functional group for conjugation is K, the conjugation is to the epsilon-amino group of K. In some embodiments, the polypeptide is X 35 The present invention further comprises a fatty acid conjugated with K, wherein the fatty acid conjugates with the epsilon-amino group of K either via a direct bond or via a linker between K and the fatty acid. In some embodiments, the amino acid having a functional group available for conjugation is γE in X1, where the conjugation is via an amide bond between the alpha-amino group of γE and the carboxyl group of the fatty acid. In some embodiments, the means for increasing the half-life of the polypeptide is a fatty acid, wherein the fatty acid conjugates with the polypeptide in an amino acid having a functional group available for conjugation, either via a direct bond or via a linker between the amino acid and the fatty acid.

[0017] In some embodiments, the N-terminal amino acid is N-acylated via a fatty acid conjugated with the N-terminal amino acid, either through direct bonding or via a linker between the amino acid and the fatty acid. In some such embodiments, the linker-fatty acid is conjugated with the peptide after peptide synthesis. In other such embodiments, the linker-fatty acid is conjugated with the amino acid (e.g., E or γE in some embodiments) before the addition of the adduct to the peptide.

[0018] In some embodiments, the polypeptide conjugates with the fatty acid via a linker between the amino acid and the fatty acid, optionally. Thus, in some embodiments, the polypeptide conjugates with the fatty acid via a direct bond between the amino acid and the fatty acid. In other embodiments, the polypeptide conjugates with the fatty acid via a linker between the amino acid and the fatty acid. In some embodiments, the fatty acid is C 16 -C 22 It is a fatty acid. In some embodiments, C 16 ~C 22 The conjugation of linkers into fatty acids is called linker-fatty acid.

[0019] Saturated C for use in this specification 16 ~C 22 Examples of fatty acids include palmitic acid (hexadecenoic acid) (C 16 (C1), hexadecanedioic acid (C1) 16 Diacid), margaric acid (heptadecanoic acid) (C 17 (C1 acid), heptadecanedioic acid) 17 Diacid, stearic acid (C 18 (C monoacid), octadecane diacid (C) 18 Diacid), nonadecylic acid (nonadecanoic acid) (C 19 (C1 acid), nonadecanedioic acid) 19 Diacid), arachidic acid (eicosanoic acid) (C 20 (C monoacid), eicosanedioic acid (C) 20 Diacid, Heneicosylic acid (Heneicosanic acid) (C 21(C1 acid), henicosanedioic acid (C1) 21 Diacids), behenic acid (docosanoic acid) (C 22 (C monoacid), docosanedioic acid (C) 22 Examples include, but are not limited to, diacides, and also include their branched and substituted derivatives.

[0020] In certain cases, the polypeptide contains a linker. The linker may have one or more (2-[2-(2-amino-ethoxy)-ethoxy]-acetyl moieties, which are optionally combined with one or more amino acids. In cases where the linker contains at least one amino acid, the amino acid may be an E or γE amino acid residue. In some cases, the linker may contain one, two, three, four, or five E or γE amino acid residues and may have a D form. For example, the linker may contain one, two, three, or four γE amino acid residues. Alternatively, the linker may contain one to five (2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)((2-[2-(2-amino-ethoxy) The linker may contain 1 to 5 amino acid residues (e.g., E or γE amino acids) used in combination with (2-[2-(2-amino-ethoxy)-ethoxy]-acetyl) ("AEEA") or 1 to 5 εK moieties. Specifically, the linker may be a combination of 1 to 5 E or γE amino acids and 1 to 5 (2-[2-(2-amino-ethoxy)-ethoxy]-acetyl) ("AEEA") or 1 to 5 εK moieties, or a combination of 1 to 5 E or γE amino acids and 1 to 5 εK moieties. In some examples, the linker may be a combination of 1, 2, or 3 γE amino acids and 1 or 2 (2-[2-(2-amino-ethoxy)-ethoxy]-acetyl) moieties or εK moieties.

[0021] In certain embodiments, the polypeptide, or a pharmaceutically acceptable salt thereof, further comprises a linker-fatty acid, the linker-fatty acid being (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-PEPEPE-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 -CO 2、 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)-CO-(CH2) 18 -CO2H, 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)-γE 2- 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 is selected from the group consisting of -CO2H.

[0022] 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 amidated.

[0023] In one embodiment of Formula I, X1 is absent, X2 is absent, X3 is G, X4 is S, X5 is P, X6 is S, X 11 is αMeL, X 13 is V, X 15 is T, X 16 is Aib, X 17 is αMeL, X 19 is Q, X 20 is K, X 21 is I or L, X 22 is L, X 26 is R, X 27 is A, X 30 is A, X 31 is R, X 35 is K, X 37 is A, X 38 is A. In such embodiments, the polypeptide comprises a 42-amino acid sequence.

[0024] Accordingly, in some embodiments, there is provided herein a polypeptide of Formula Ia, or a pharmaceutically acceptable salt thereof, wherein GSPSIVTS αMeL DVPT Aib αMeL LQKX 21 LEQERAEKARQQAKEAAEILAQV (comprises SEQ ID NO: 69), X 21 is I or L, [when X1 and X2 are absent, X 21 is at position 19 of the polypeptide], there is provided a polypeptide, or a pharmaceutically acceptable salt thereof, wherein the N-terminal amino acid is optionally N-acetylated, and the C-terminal amino acid is optionally amidated.

[0025] In some embodiments of formula Ia, X 21 I is. In some embodiments, the polypeptide comprises SEQ ID NO: 20. In other embodiments, the polypeptide is SEQ ID NO: 20. Alternatively, the polypeptide may have at least about 90 to about 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.

[0026] In certain embodiments of formula Ia, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 21 is I, and the polypeptide further comprises a linker-fatty acid. In certain embodiments, the linker-fatty acid is γE-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-εK-CO-(CH2) 18 -CO2H. In certain embodiments of formula Ia, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 35 In this embodiment, K conjugates with a 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 90-97% 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.

[0027] In certain embodiments of formula Ia, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 21 L is the polypeptide, and the polypeptide further comprises a linker-fatty acid. In certain embodiments, the linker-fatty acid is γE-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-εK-CO-(CH2) 18 -CO2H. In certain embodiments of formula Ia, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 35In this embodiment, K conjugates with a 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 90-97% 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.

[0028] In certain embodiments of formula Ia, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 21 is I, and the polypeptide further comprises a linker-fatty acid. In certain embodiments, the linker-fatty acid is 2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-PKE-γE-CO-(CH2) 18 -CO2H. In certain embodiments of formula Ia, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 35 In this embodiment, K conjugates with a 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 90-97% 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.

[0029] In certain embodiments of formula Ia, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 21 is I, and the polypeptide further comprises a linker-fatty acid. In certain embodiments, the linker-fatty acid is Ahx-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-γE-CO-(CH2) 18 -CO2H. In certain embodiments of formula Ia, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 35In this embodiment, K conjugates with a linker-fatty acid. In another embodiment, the polypeptide comprises SEQ ID NO: 4. Alternatively, the polypeptide may have at least about 90-97% 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.

[0030] In certain embodiments of formula Ia, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 21 is I, and the polypeptide further comprises a linker-fatty acid. In certain embodiments, the linker-fatty acid is γE-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-Ahx-CO-(CH2) 18 -CO2H. In certain embodiments of formula Ia, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 35 In this embodiment, K conjugates with a 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 90-97% 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.

[0031] In certain embodiments of formula Ia, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 21 is I, and the polypeptide further comprises a linker-fatty acid. In certain embodiments, the linker-fatty acid is γE-GKEKEKE-CO-(CH2) 18 -CO2H. In certain embodiments of formula Ia, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 35In this embodiment, K conjugates with a 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 90-97% 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.

[0032] In certain embodiments of formula Ia, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 21 is I, and the polypeptide further comprises a linker-fatty acid. In certain embodiments, the linker-fatty acid is G-(βA)3-γE-CO-(CH2) 18 -CO2H. In certain embodiments of formula Ia, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 35 In this embodiment, K conjugates with a 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 90-97% 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.

[0033] In certain embodiments of formula Ia, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 21 is I, and the polypeptide further comprises a linker-fatty acid. In certain embodiments, the linker-fatty acid is G-(βA)4-γE-CO-(CH2) 18 -CO2H. In certain embodiments of formula Ia, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 35In this embodiment, K conjugates with a 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 90-97% 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.

[0034] In certain embodiments of formula Ia, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 21 is I, and the polypeptide further comprises a linker-fatty acid. In certain embodiments, the linker-fatty acid is 2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-KEKEKE-γE-CO-(CH2) 18 -CO2H. In certain embodiments of formula Ia, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 35 In this embodiment, K conjugates with a 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 90-97% 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.

[0035] In certain embodiments of formula Ia, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 21 is I, and the polypeptide further comprises a linker-fatty acid. In certain embodiments, the linker-fatty acid is βA-S-βA-S-γE-CO-(CH2) 18 -CO2H. In certain embodiments of formula Ia, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 35In this embodiment, K conjugates with a 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 90-97% 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.

[0036] In certain embodiments of formula Ia, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 21 is I, and the polypeptide further comprises a linker-fatty acid. In certain embodiments, the linker-fatty acid is γE-PKE-Ahx-CO-(CH2) 18 -CO2H. In certain embodiments of formula Ia, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 35 In this embodiment, K conjugates with a 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 90-97% 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.

[0037] In certain embodiments of formula Ia, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 21 L is the polypeptide, and the polypeptide further comprises a linker-fatty acid. In certain embodiments, the linker-fatty acid is γE-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-εK-CO-(CH2) 16 -CO2H. In certain embodiments of formula Ia, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 35In this embodiment, K conjugates with a 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 90-97% 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.

[0038] In certain embodiments of formula Ia, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 21 is I, and the polypeptide further comprises a linker-fatty acid. In certain embodiments, the linker-fatty acid is γE-PKE-εK-CO-(CH2) 18 -CO2H. In certain embodiments of formula Ia, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 35 In this case, K conjugates with the linker-fatty acid. Alternatively, the polypeptide may have at least about 90-97% 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.

[0039] In certain embodiments of formula Ia, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 21 is I, and the polypeptide further comprises a linker-fatty acid. In certain embodiments, the linker-fatty acid is γE-PPP-εK-CO-(CH2) 18 -CO2H. In certain embodiments of formula Ia, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 35In this embodiment, K conjugates with a 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 90-97% 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.

[0040] In certain embodiments of formula Ia, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 21 is I, and the polypeptide further comprises a linker-fatty acid. In certain embodiments, the linker-fatty acid is γE-PPPPPP-CO-(CH2) 18 -CO2H. In certain embodiments of formula Ia, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 35 In this embodiment, K conjugates with a 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 90-97% 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.

[0041] In certain embodiments of formula Ia, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 21 is I, and the polypeptide further comprises a linker-fatty acid. In certain embodiments, the linker-fatty acid is γE-PEPEPE-CO-(CH2) 18 -CO2H. In certain embodiments of formula Ia, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 35In this embodiment, K conjugates with a 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 90-97% 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.

[0042] In certain embodiments of formula Ia, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 21 is I, and the polypeptide further comprises a linker-fatty acid. In certain embodiments, the linker-fatty acid is γE-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)2-εK-CO-(CH2) 18 -CO2H. In certain embodiments of formula Ia, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 35 In this embodiment, K conjugates with a 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 90-97% 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.

[0043] In certain embodiments of formula Ia, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 21 is I, and the polypeptide further comprises a linker-fatty acid. In certain embodiments, the linker-fatty acid is γE-εK-γE-εK-CO-(CH2) 18 -CO2H. In certain embodiments of formula Ia, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 35In this case, K conjugates with the linker-fatty acid. Alternatively, the polypeptide may have at least approximately 90-97% 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.

[0044] In certain embodiments of formula Ia, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 21 is I, and the polypeptide further comprises a linker-fatty acid. In certain embodiments, the linker-fatty acid is γE-εK-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-εK-CO-(CH2) 18 -CO2H. In certain embodiments of formula Ia, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 35 In this embodiment, K conjugates with a 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 90-97% 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.

[0045] In certain embodiments of formula Ia, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 21 is I, and the polypeptide further comprises a linker-fatty acid. In certain embodiments, the linker-fatty acid is γE-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-εK-CO-(CH2) 16 -CO2H. In certain embodiments of formula Ia, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 35In this embodiment, K conjugates with a 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 90-97% 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.

[0046] In another embodiment of formula I, X1 is Pyr, X2 is G, X3 is S, X4 is P, X5 is S, X6 is G, X 11 αMeL and X 13 V is X 15 is T, and X 16 Aib and X 17 αMeL and X 19 Q is X 20 is K, and X 21 is I and X 22 L is X 26 R is X 27 A is X 30 A is X 31 R is X 35 is K and X 37 A is X 38 This is A. In such embodiments, the polypeptide comprises a 44-amino acid sequence.

[0047] Therefore, in some embodiments, this specification refers to a polypeptide of formula Ib, or a pharmaceutically acceptable salt thereof, Pyr GSPSGIVTS αMeL DVPT Aib αMeL LQKILEQERAEKARQQAKEAAEILAQV (Sequence ID 70) is included, Polypeptides, or pharmaceutically acceptable salts thereof, are provided, in which the C-terminal amino acid is optionally amidated.

[0048] In another embodiment, the polypeptide comprises SEQ ID NO: 23. Alternatively, the polypeptide may have at least about 90-97% 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.

[0049] In certain embodiments of formula Ib, comprising a polypeptide or a pharmaceutically acceptable salt thereof, the polypeptide further comprises a linker-fatty acid. In certain embodiments, the linker-fatty acid is γE-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-εK-CO-(CH2) 18 -CO2H. In certain embodiments of formula Ib, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 35 In this embodiment, K conjugates with a 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 90-97% 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.

[0050] In certain embodiments of formula Ib, comprising a polypeptide or a pharmaceutically acceptable salt thereof, the polypeptide further comprises a linker-fatty acid. In certain embodiments, the linker-fatty acid is γE-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-εK-CO-(CH2) 16 -CO2H. In certain embodiments of formula Ib, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 35In this embodiment, K conjugates with a 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 90-97% 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.

[0051] In another embodiment of formula I, X1 is γE, E, or absent, X2 is absent, X3 is G, X4 is S, X5 is P, X6 is S, X 11 αMeL and X 13 V is X 15 is T, and X 16 Aib and X 17 αMeL and X 19 is Q or E, and X 20 is K and X 21 is I and X 22 L is X 26 R is X 27 A is X 30 A is X 31 R is X 35 is T, and X 37 A is X 38 A is the answer.

[0052] In another embodiment of formula I, X1 is absent, X2 is absent, X3 is G, X4 is S, X5 is P, X6 is S, X 11 αMeL and X 13 V is X 15 is T, and X 16 Aib and X 17 αMeL and X 19 Q is X 20 is K and X 21 is I and X 22 L is X 26 R is X27 A is X 30 A is X 31 R is X 35 is T, and X 37 A is X 38 This is A. In such embodiments, the polypeptide comprises a 42-amino acid sequence.

[0053] In another embodiment, the polypeptide comprises SEQ ID NO: 27. Alternatively, the polypeptide may have at least about 90-97% 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.

[0054] 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, X 11 αMeL and X 13 V is X 15 is T, and X 16 Aib and X 17 αMeL and X 19 is Q or E, and X 20 is K and X 21 is I and X 22 L is X 26 R is X 27 A is X 30 A is X 31 R is X 35 is T, and X 37 A is X 38 This is A. In such embodiments, the polypeptide comprises a 43-amino acid sequence.

[0055] Therefore, in some embodiments, this specification refers to polypeptides of formula Ic, or pharmaceutically acceptable salts thereof, X1GSPSIVTS αMeL DVPT Aib αMeLLX19 Includes KILEQERAEKARQQATEAAEILAQV (Sequence ID 71), X1 is γE or E, X 19 However, if Q or E, and [X2 does not exist, X 19 It is ranked 18th among polypeptides. Polypeptides, or pharmaceutically acceptable salts thereof, are provided, in which the C-terminal amino acid is optionally amidated.

[0056] In certain embodiments of formula Ic, which comprises a polypeptide or a pharmaceutically acceptable salt thereof, X1 is γE, and X 19 Q is the linker-fatty acid, 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 90-97% 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.

[0057] In certain embodiments of formula Ic, which comprises a polypeptide or a pharmaceutically acceptable salt thereof, X1 is γE, and X 19 Q is the linker-fatty acid, 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 90-97% 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.

[0058] In certain embodiments of formula Ic, which comprises a polypeptide or a pharmaceutically acceptable salt thereof, X1 is γE, and X 19 Q is the linker-fatty acid, 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 90-97% 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.

[0059] In certain embodiments of formula Ic, which comprises a polypeptide or a pharmaceutically acceptable salt thereof, X1 is γE, and X 19 E is the linker-fatty acid, (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 90-97% 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.

[0060] In another embodiment of formula 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, X 11 αMeL and X 13 It is D-Val, and X 15 is T or I, and X 16 is Aib or G, and X 17 αMeL and X 19 Q is X 20 is I and X 21 is K and X 22 L is X26 R is X 27 A is X 30 A is X 31 R is X 35 is K and X 37 N is X 38 This is T. In such embodiments, the polypeptide comprises a 38-amino acid sequence.

[0061] Therefore, in some embodiments, this specification refers to a polypeptide of formula Id, or a pharmaceutically acceptable salt thereof, IVTS αMeL D D-Val PX 15 X 16 Contains αMeL LQIKLEQERAEKARQQAKENTEILAQV (Sequence ID 72), X 15 However, if T or I and [X1~X6 does not exist, X 15 It is ranked 9th among polypeptides. X 16 However, if G or Aib and [X1~X6 does not exist, X 16 It is ranked 10th among polypeptides. A polypeptide or a pharmaceutically acceptable salt thereof is provided, wherein the N-terminus is an amino acid, optionally N-acetylated, and the C-terminal amino acid is optionally amidated.

[0062] In certain embodiments of formula Id, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 15 is T, and X 16 The polypeptide is G, and further comprises a linker-fatty acid. In certain embodiments, the linker-fatty acid is 2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)2-γE-CO-(CH2) 18 -CO2H. In certain embodiments of formula Id, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 35In this embodiment, K conjugates with a linker-fatty acid. In another embodiment, the polypeptide is SEQ ID NO: 29. Alternatively, the polypeptide may have at least about 90-97% 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.

[0063] In certain embodiments of formula Id, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 15 is T, and X 16 The polypeptide is G, and further comprises a linker-fatty acid. In certain embodiments, the linker-fatty acid is γE-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-γE-CO-(CH2) 18 -CO2H. In certain embodiments of formula Id, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 35 In this embodiment, K conjugates with a 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 90-97% 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.

[0064] In certain embodiments of formula Id, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 15 is T, and X 16 The polypeptide is G, and further comprises a linker-fatty acid. In certain embodiments, the linker-fatty acid is γE2-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-CO-(CH2) 18 -CO2H. In certain embodiments of formula Id, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 35In this embodiment, K conjugates with a 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 90-99% sequence similarity to SEQ ID NO: 31. Alternatively, the polypeptide may have at least about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence similarity to SEQ ID NO: 31.

[0065] In certain embodiments of formula Id, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 15 is T, and X 16 The polypeptide is G, and further comprises a linker-fatty acid. In certain embodiments, the linker-fatty acid is 2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-εK-γE-CO-(CH2) 18 -CO2. In certain embodiments of formula Id, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 35 In this embodiment, K conjugates with a 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 90-97% 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.

[0066] In certain embodiments of formula Id, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 15 is T, and X 16 is G, and the polypeptide further comprises a linker-fatty acid. In certain embodiments, the linker-fatty acid is 2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-γE-εK-CO-(CH2) 18 -CO2H. In certain embodiments of formula Id, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 35In this embodiment, K conjugates with a 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 90-97% 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.

[0067] In certain embodiments of formula Id, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 15 is T, and X 16 The polypeptide is G, and further comprises a linker-fatty acid. In certain embodiments, the linker-fatty acid is εK-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-γE-CO-(CH2) 18 -CO2H. In certain embodiments of polypeptides or pharmaceutically acceptable salts thereof, X 35 In this embodiment, K conjugates with a linker-fatty acid. In another embodiment, the polypeptide comprises SEQ ID NO: 34. Alternatively, the polypeptide may have at least about 90-97% 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.

[0068] In certain embodiments of formula Id, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 15 is T, and X 16 The polypeptide is G, and further comprises a linker-fatty acid. In certain embodiments, the linker-fatty acid is εK-γE-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-CO-(CH2) 18 -CO2H. In certain embodiments of formula Id, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 35In this embodiment, K conjugates with a 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 90-97% 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.

[0069] In certain embodiments of formula Id, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 15 is T, and X 16 The polypeptide is G, and further comprises a linker-fatty acid. In certain embodiments, the linker-fatty acid is γE-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-εK-CO-(CH2) 18 -CO2H. In certain embodiments of formula Id, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 35 In this embodiment, K conjugates with a 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 90-97% 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.

[0070] In certain embodiments of formula Id, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 15 is T, and X 16 The polypeptide is G, and further comprises a linker-fatty acid. In certain embodiments, the linker-fatty acid is γE-εK-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-CO-(CH2) 18 -CO2H. In certain embodiments of formula Id, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 35In this embodiment, K conjugates with a 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 90-97% 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.

[0071] In certain embodiments of formula Id, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 15 is T, and X 16 is G, and the polypeptide further comprises a linker-fatty acid. In certain embodiments, the linker-fatty acid is γE-γE-εK-CO-(CH2) 18 -CO2H. In certain embodiments of formula Id, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 35 In this embodiment, K conjugates with a 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 90-97% 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.

[0072] In certain embodiments of formula Id, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 15 is T, and X 16 is G, and the polypeptide further comprises a linker-fatty acid. In certain embodiments, the linker-fatty acid is γE-εK-γE-CO-(CH2) 18 -CO2H. In certain embodiments of formula Id, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 35In this embodiment, K conjugates with a 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 90-97% 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.

[0073] In certain embodiments of formula Id, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 15 is T, and X 16 is G, and the polypeptide further comprises a linker-fatty acid. In certain embodiments, the linker-fatty acid is εK-γE-γE-CO-(CH2) 18 -CO2H. In certain embodiments of formula Id, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 35 In this embodiment, K conjugates with a 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 90-97% 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.

[0074] In certain embodiments of formula Id, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 15 is T, and X 16 Aib is the polypeptide, and the polypeptide further comprises a linker-fatty acid. In certain embodiments, the linker-fatty acid is 2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)2-γE-CO-(CH2) 18 -CO2H. In certain embodiments of formula Id, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 35In this embodiment, K conjugates with a 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 90-97% 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.

[0075] In certain embodiments of formula Id, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 15 is I and X 16 Aib is the polypeptide, and the polypeptide further comprises a linker-fatty acid moiety. In certain embodiments, the linker-fatty acid is 2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)2-γE-CO-(CH2) 18 -CO2H. In certain embodiments of formula Id, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 35 In this embodiment, K conjugates with a 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 90-97% 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.

[0076] In certain embodiments of formula Id, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 15 is T, and X 16 is G, and the polypeptide further comprises a linker-fatty acid. In certain embodiments, the linker-fatty acid is GKEKEKE-CO-(CH2) 18 -CO2H. In certain embodiments of formula Id, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 35In this embodiment, K conjugates with a 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 90-97% 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.

[0077] In certain embodiments of formula Id, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 15 is T, and X 16 The polypeptide is G, and further comprises a linker-fatty acid. In certain embodiments, the linker-fatty acid is 2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-γE 2- CO-(CH2) 18 -CO2H. In certain embodiments of formula Id, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 35 In this embodiment, K conjugates with a 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 90-97% 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.

[0078] In certain embodiments of formula Id, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 15 is I and X 16 is Aib, and the polypeptide further comprises a linker-fatty acid. In certain embodiments, the linker-fatty acid is γE2-CO-(CH2) 18 -CO2H. In certain embodiments of formula Id, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 35In this embodiment, K conjugates with a 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 90-97% 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.

[0079] In another embodiment of Equation I, X1 does not exist, X2 does not exist, X3 does not exist, X4 is P, X5 is P, X6 is P, X 11 αMeL and X 13 is V or D-Val, and X 15 is T, and X 16 Aib and X 17 αMeL and X 19 Q is X 20 is K and X 21 is L or Aib, and X 22 L is X 26 R is X 27 A is X 30 A is X 31 R is X 35 is K and X 37 is A or N, and X 38 is A or T. In such embodiments, the polypeptide comprises a 41-amino acid sequence.

[0080] Therefore, in some embodiments, this specification refers to polypeptides of formula Ie, or pharmaceutically acceptable salts thereof, PPPIVTS αMeL DX 13 PTAib αMeL LQKX 21 LEQERAEKARQQAKEX 37 X 38 Includes EILAQV (Sequence ID 73), X 13 However, if V or D-Val and [X1~X3 does not exist, X 13It is ranked 10th among polypeptides. X 21 However, if L or Aib and [X1~X3 does not exist, X 21 It is ranked 18th among polypeptides. X 37 However, if A or N and [X1~X3 does not exist, then X 37 It is ranked 34th among polypeptides. X 38 However, if A or T and [X1~X3 does not exist, X 38 It is ranked 35th among polypeptides. A polypeptide, or a pharmaceutically acceptable salt thereof, is provided, in which the N-terminal amino acid is optionally N-acetylated and the C-terminal amino acid is optionally amidated.

[0081] In certain embodiments of formula Ie, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 13 It is D-Val, and X 21 Aib and X 37 N is X 38 is T, and the polypeptide further comprises a linker-fatty acid. In certain embodiments, the linker-fatty acid is γE-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-εK-CO-(CH2) 18 -CO2H. In certain embodiments, X 35 In this case, K conjugates with a 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 90-97% 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.

[0082] In certain embodiments of formula Id, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 13 It is D-Val, and X 21 L is X37 A is X 38 A is the polypeptide, and the polypeptide further comprises a linker-fatty acid. In certain embodiments, the linker-fatty acid is γE-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-εK-CO-(CH2) 18 -CO2H. In certain embodiments of formula Id, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 35 In this case, K conjugates with a 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 90-97% 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.

[0083] In certain embodiments of formula Id, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 13 V is X 21 L is X 37 A is X 38 A is the polypeptide, and the polypeptide further comprises a linker-fatty acid. In certain embodiments, the linker-fatty acid is (γE-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-εK-CO-(CH2) 18 -CO2H. In certain embodiments of formula Id, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 35 In this case, K conjugates with a 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 90-97% 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.

[0084] In another embodiment of formula I, X1 is γE, X2 is absent, X3 is absent, X4 is G, X5 is P, X6 is S, X 11 αMeL and X 13 It is D-Val, and X 15 is I and X 16 Aib and X 17 αMeL and X 19 Q is X 20 is I and X 21 is K and X 22 L is X 26 R is X 27 A is X 30 A is X 31 R is X 35 is K and X 37 N is X 38 is T. In certain embodiments, the polypeptide further comprises a linker-fatty acid. In certain embodiments, the linker-fatty acid is (2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)2-γE-CO-(CH2) 18 -CO2H. In certain embodiments, which include polypeptides or pharmaceutically acceptable salts thereof, X 35 In this case, K conjugates with linker-fatty acids.

[0085] In some embodiments, the polypeptide comprises SEQ ID NO: 45. Alternatively, the polypeptide may have at least about 90-97% 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.

[0086] In another embodiment of formula 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, X 11 L is X 13 V is X 15 is I and X16 G is X 17 αMeL and X 19 Q is X 20 is K, and K 21 L is X 22 is I and X 26 is K and X 27 Q is X 30 E is X 31 is K and X 35 is K and X 37 N is X 38 It is T.

[0087] In certain embodiments of the polypeptide, or a pharmaceutically acceptable salt thereof, the polypeptide further comprises a linker-fatty acid. In certain embodiments, the linker-fatty acid is (2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)2γE-C2O-OH). In certain embodiments comprising the polypeptide, or a pharmaceutically acceptable salt thereof, X 35 In this case, K conjugates with linker-fatty acids.

[0088] In some embodiments, the polypeptide comprises SEQ ID NO: 49. Alternatively, the polypeptide may have at least about 90-97% 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.

[0089] In another embodiment of formula 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, X 11 L is X 13 V is X 15 is I and X 16 G is X 17 αMeL and X 19 Q is X 20 is K, and K 21L is X 22 L is X 26 R is X 27 A is X 30 A is X 31 R is X 35 is K and X 37 A is X 38 A is the answer.

[0090] In certain embodiments of the polypeptide, or a pharmaceutically acceptable salt thereof, the polypeptide further comprises a linker-fatty acid. In certain embodiments, the linker-fatty acid is 2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)2-γE-CO-(CH2) 18 -CO2H. In certain embodiments, which include polypeptides or pharmaceutically acceptable salts thereof, X 35 In this case, K conjugates with linker-fatty acids.

[0091] 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 90–97% 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.

[0092] In one embodiment, the Specified herein provides a polypeptide or a pharmaceutically acceptable salt thereof comprising a sequence having at least eight amino acid modifications from human urocortin-2 (SEQ ID NO: 53). In some embodiments, the polypeptides provided herein comprise a sequence containing at least one amino acid modification at positions 3, 19, 22, 23, 26, 30, 33, or 37 from human urocortin-2 (SEQ ID NO: 53). In some embodiments, the polypeptides provided herein comprise a sequence containing at least one of the following amino acid modifications from human urocortin-2 (SEQ ID NO: 53): L3T, A19E, R22E, A23K, E26Q, T30E, R33E, R37Q, or any combination thereof.

[0093] In some embodiments, the Specified Reference Numerals (SEQ ID NOs) 1-52 are provided as polypeptides comprising any one of SEQ ID NOs) 1-52, or as pharmaceutically acceptable salts thereof. Alternatively, the polypeptides may have at least about 90-99% sequence similarity to any one of SEQ ID NOs) 1-52, or as pharmaceutically acceptable salts thereof. Alternatively, the polypeptides may have at least about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence similarity to any one of SEQ ID NOs) 1-52.

[0094] In certain embodiments, the polypeptide comprises SEQ ID NO: 1 or a pharmaceutically acceptable salt thereof. In certain embodiments, the polypeptide comprises SEQ ID NO: 2 or a pharmaceutically acceptable salt thereof. In certain embodiments, the polypeptide comprises SEQ ID NO: 22 or a pharmaceutically acceptable salt thereof. In certain embodiments, the polypeptide comprises SEQ ID NO: 25 or a pharmaceutically acceptable salt thereof. In certain embodiments, the polypeptide comprises SEQ ID NO: 29 or a pharmaceutically acceptable salt thereof. In certain embodiments, the polypeptide comprises SEQ ID NO: 41 or a pharmaceutically acceptable salt thereof. In certain embodiments, the polypeptide comprises SEQ ID NO: 42 or a pharmaceutically acceptable salt thereof. In certain embodiments, the polypeptide comprises SEQ ID NO: 45 or a pharmaceutically acceptable salt thereof. In certain embodiments, the polypeptide comprises SEQ ID NO: 46 or a pharmaceutically acceptable salt thereof. In certain embodiments, the polypeptide comprises SEQ ID NO: 48 or a pharmaceutically acceptable salt thereof. In certain embodiments, the polypeptide comprises SEQ ID NO: 49 or a pharmaceutically acceptable salt thereof. In certain embodiments, the polypeptide comprises SEQ ID NO: 50 or a pharmaceutically acceptable salt thereof.

[0095] In certain embodiments, the polypeptide, or a pharmaceutically acceptable salt thereof, is selected from the group consisting of SEQ ID NOs: 1, 2, 22, 25, 29, 41, 42, 45, 46, 48, 49, and 50.

[0096] In another embodiment, the specification provides a pharmaceutical composition comprising a polypeptide or a pharmaceutically acceptable salt thereof and at least one pharmaceutically acceptable carrier, diluent, or excipient.

[0097] In another embodiment, a method is provided for treating a disease or condition selected from the group consisting of diabetes mellitus, 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), comprising administering an effective amount of a polypeptide, or a pharmaceutically acceptable salt thereof, to an individual in need thereof.

[0098] In another embodiment, this specification provides polypeptides or pharmaceutically acceptable salts thereof for use in therapy.

[0099] In another embodiment, the Specified Reference Indicators provide polypeptides or pharmaceutically acceptable salts thereof for use in the treatment of diseases or conditions selected from the group consisting of diabetes mellitus, 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).

[0100] In one embodiment, the disclosure provides the use of polypeptides or pharmaceutically acceptable salts thereof in the manufacture of pharmaceuticals for the treatment of diseases or conditions selected from the group consisting of diabetes mellitus, 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 Specified Provision provides a method for treating a disease or condition selected from the group consisting of diabetes mellitus, 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), comprising administering to an individual in need thereof an effective amount of a polypeptide, or a pharmaceutically acceptable salt thereof, and an effective amount of an additional therapeutic agent.

[0102] In one embodiment, the Specified herein provides polypeptides, or pharmaceutically acceptable salts thereof, that can be used simultaneously with, separately or in succession in combination with, one or more additional therapeutic agents.

[0103] In one embodiment, the additional therapeutic agent is a GIP and GLP-1 receptor dual agonist. In a particular embodiment, the GIP and GLP-1 receptor dual agonist is a compound comprising SEQ ID NO: 55. In one embodiment, the additional therapeutic agent is a GLP-1 receptor agonist. 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. [Modes for carrying out the invention]

[0104] Amino acids contain an amino group and a carboxylic acid group. In α-amino acids, the amino group and carboxylic acid group, including the 20 standard amino acids, are covalently bonded to the same carbon atom defined as the α-carbon. The α-carbon may also be substituted with additional functional groups, which may have defined stereochemistry. The 20 standard amino acids are most often found as L-stereoisomers, and therefore, unless otherwise specified, references to amino acids are assumed to refer specifically to their L-stereoisomers.

[0105] Polypeptides are polymers of amino acids covalently linked via amide bonds between a carboxylic acid group on one monomer and an amino group on the next monomer. Most commonly, these are α-carboxylic acid and α-amino groups, respectively, and therefore, unless otherwise specified, polypeptides written using standard one- or three-letter amino acid codes are assumed to be α-amino acid polymers.

[0106] Typically, amino acids are depicted with the amino group on the left and the carboxylic acid group on the right, and polypeptide sequences are written such that, if they are unmodified linear alpha-amino acid chains, the first amino acid is assumed to have a free amino group on the left end of the chain (also written as H2N-polypeptide, H-polypeptide, or polypeptide), and the last amino acid is assumed to have a free carboxylic acid group at the right end of the chain (polypeptide-COOH, typically written as polypeptide or polypeptide-OH). Thus, the left end of a polypeptide is also known as the N-terminus (due to the nitrogen in the amine group), and the right end of a polypeptide is also known as the C-terminus (due to the carboxylic acid group). Unless otherwise specified, if a polypeptide is simply described as a standard one- or three-letter amino acid code, it is assumed that the polypeptide begins with a free amino group at the N terminus and ends with a free carboxylic acid at the C terminus.

[0107] Polypeptides may further include N-terminal modifications (e.g., acetylation, methylation, acylation, or other modifications). Polypeptides may also have C-terminal modifications, most commonly the C-terminal group being an amide (-CONH2, usually written as polypeptide-NH2) instead of a carboxylic acid, although other C-terminal modifications are also known in the art.

[0108] The amino acid sequences of the polypeptides described herein typically incorporate naturally occurring amino acids, as well as non-natural amino acids such as α-methyl-substituted residues of natural amino acids (e.g., α-methylleucine (αMeL)) and α-aminoisobutyric acid (Aib), as shown herein using standard single-letter codes (e.g., L = leucine). The structures and modifications of these amino acids and other monomers are shown below. [ka]

[0109] As a result, as used herein, "Ac" means N-acetyl. As used herein, "Pyr" means L-pyroglutamic acid. As used herein, "Aib" means 2-aminoisobutyric acid. As used herein, "D-Val" means D- Means valine. As used herein, "Orn" means L-ornithine. As used herein, "Dap" means L-2,3-diaminopropionic acid. As used herein, "αMeL" means alpha-methyl-L-leucine. As used herein, "αMeF" means alpha-methyl-L-phenylalanine. As used herein, "NMeI" means N-methyl-L-isoleucine. As used herein, "NMeN" means N-methyl-L-asparagine. As used herein, "NMeD" means N-methyl-L-aspartic acid. As used herein, "βA" means beta-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 epsilon-L-lysine. As used herein, "γE" means gamma-L-glutamic acid.

[0110] "N-term" can refer to an N-terminus or N-terminal (when used as a modifier). "C-term" can refer to a C-terminus or C-terminal (when used as a modifier).

[0111] It should be noted that the beneficial combinations of properties of the exemplary analogues described herein are not the result of any single modification alone, but rather are achieved through novel combinations of the structural features described herein.

[0112] In some embodiments, the present disclosure relates to a polypeptide of formula I, or a pharmaceutically acceptable salt thereof. X1X2X3X4X5X6IVTSX 11 DX 13 PX 15 X 16 X 17 LX19 X 20 X 21 X 22 EQEX 26 X 27 EKX 30 X 31 QQAX 35 EX 37 X 38 Includes EILAQV (Sequence ID 68), X1 is either Pyr, E, γE, or absent. X2 is either G or does not exist. X3 is either G, S, or does not exist. X4 is either S, P, G, or does not exist. X5 is either S, P, or does not exist. X6 is either G, S, or P, or does not exist. X 11 However, it is L or αMeL, X 13 However, it is V or D-Val, X 15 However, it is T or I, X 16 However, it is Aib or G, X 17 However, it is L or αMeL, X 19 However, it is either Q or E, X 20 However, it is K or I, X 21 However, it is I, K, Aib, or L. X 22 However, it is either I or L. X 26 However, it is R or K, X 27 However, it is either A or Q, X 30 However, it is A or E, X 31 However, it is R or K, X 35 However, it is either K or T, X 37 However, it is either A or N, X 38 However, it is A or T, Polypeptides or pharmaceutically acceptable salts thereof are provided, wherein the N-terminal amino acid is optionally N-acylated, N-acetylated, or N-methylated, and the C-terminal amino acid is optionally amidated.

[0113] In some embodiments of Formula I, the V at position 44 is the C-terminal amino acid. The C-terminus may contain an amide, carboxylic acid, or other modification. The N-terminal amino acid is embodiment-dependent but is always the first amino acid present in the sequence that indicates 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; acylation, or other modifications). Furthermore, if any amino acid at positions X1-X6 is indicated as "absent," the next available amino acid conjugates with the previously available amino acid. Thus, various embodiments of Formula I result in polypeptides of varying lengths, e.g., 38-44 amino acids.

[0114] Furthermore, in addition to the sequences described herein, the polypeptides described herein may contain one or more conservative amino acid substitutions, provided that the polypeptide remains capable of being active in CRHR2.

[0115] In some embodiments, the polypeptide, or a pharmaceutically acceptable salt thereof, further includes means for increasing the half-life of the polypeptide. In some embodiments, the polypeptide, or a pharmaceutically acceptable salt thereof, further includes means for increasing the half-life of the polypeptide via any amino acid on the polypeptide having a functional group suitable for conjugation. In some embodiments, the amino acid having a functional group suitable for conjugation is γE, E, or K.

[0116] In some embodiments, the functional groups available for conjugation conjugate with phosphonates, tetrazoles, 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, fatty acids, linkers, proteins, or protein fragments may further act as albumin binders, providing the potential to generate longer-acting compounds.

[0117] In some embodiments, these means are fatty acids.

[0118] In some embodiments, the amino acid having a functional group for conjugation is K, the conjugation is to the epsilon-amino group of K. In some embodiments, the polypeptide is X 35 The present invention further comprises a fatty acid conjugated with K, wherein the fatty acid is conjugated to the epsilon-amino group of K either via direct bonding or via a linker between K and the fatty acid. In some embodiments, the amino acid having a functional group for conjugation is γE in X1, where the conjugation is via an amide bond between the alpha-amino group of γE and the carboxyl group of the fatty acid. In some embodiments, the means for increasing the half-life of the polypeptide is a fatty acid, where the fatty acid conjugates with the polypeptide in an amino acid having a functional group available for conjugation, either via direct bonding or via a linker between the amino acid and the fatty acid.

[0119] In some embodiments, the N-terminal amino acid is N-acylated via a fatty acid, via a fatty acid conjugated with the N-terminal amino acid, via direct bonding, or via a linker between the amino acid and the fatty acid. In some such embodiments, the linker-fatty acid is conjugated with the peptide after peptide synthesis. In other such embodiments, the linker-fatty acid is conjugated with the amino acid (e.g., E or γE in some embodiments) before addition to the peptide.

[0120] In some embodiments, the polypeptide conjugates with the fatty acid via a linker between the amino acid and the fatty acid, optionally. Thus, in some embodiments, the polypeptide conjugates with the fatty acid via a direct bond between the amino acid and the fatty acid. In other embodiments, the polypeptide conjugates with the fatty acid via a linker between the amino acid and the fatty acid. In some embodiments, the fatty acid is C 16 ~C 22 It is a fatty acid. In some embodiments, C 16 ~C 22 The conjugation of linkers into fatty acids is called linker-fatty acid.

[0121] Saturated C for use in this specification 16 ~C 22 Examples of fatty acids include palmitic acid (hexadecenoic acid) (C 16 (C1), hexadecanedioic acid (C1) 16 Diacid), margaric acid (heptadecanoic acid) (C 17 (C1 acid), heptadecanedioic acid) 17 Diacid, stearic acid (C 18 (C monoacid), octadecane diacid (C) 18 Diacid), nonadecylic acid (nonadecanoic acid) (C 19 (C1 acid), nonadecanedioic acid) 19 Diacid), arachidic acid (eicosanoic acid) (C 20 (C monoacid), eicosanedioic acid (C) 20 Diacid, Heneicosylic acid (Heneicosanic acid) (C 21(C1 acid), henicosanedioic acid (C1) 21 Diacids), behenic acid (docosanoic acid) (C 22 (C monoacid), docosanedioic acid (C) 22 Examples include, but are not limited to, diacides, and also include their branched and substituted derivatives.

[0122] In certain cases, the linker may have one or more (2-[2-(2-amino-ethoxy)-ethoxy]-acetyl) moieties, which are optionally combined with one or more amino acids.

[0123] In examples where the linker contains at least one amino acid, the amino acid can be one to five E or γE amino acid residues. In some examples, the linker may contain one, two, three, four, or five E or γE amino acid residues and may include its D form. For example, the linker may contain one, two, three, or four γE amino acid residues. Alternatively, the linker may contain one to five (2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)((2-[2-(2-amino-ethoxy)-ethoxy]-acetyl), "AEEA") or one to five amino acid residues (e.g., E or γE amino acids) used in combination with one to five εK moieties. Specifically, the linker may be a combination of one to five E or γE amino acids and 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 and one to five εK moieties. In some examples, the linker may be a combination of one, two, or three γGlu amino acids and one or two (2-[2-(2-amino-ethoxy)-ethoxy]-acetyl) or εK moieties.

[0124] In certain embodiments, the polypeptide, or a pharmaceutically acceptable salt thereof, further comprises a linker-fatty acid, the linker-fatty acid being (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-PEPEPE-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 -CO 2、 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)-CO-(CH2) 18 -CO2H,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)-γE 2- CO-(CH2) 18 -CO2H,γE-(2-[2-(2-amino-ethoxy)-ethoxy]acetyl)-εK-CO-(CH2) 18-CO2H, 2-[2-(2-aminoethoxy)-ethoxy]-acetyl)2γE-C20-OH, and γE-(2-[2-(2-aminoethoxy)-ethoxy]-acetyl)-εK-CO-(CH2) 18 Selected from the group consisting of -CO2H.

[0125] 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.

[0126] In one embodiment of formula I, X1 is absent, X2 is absent, X3 is G, X4 is S, X5 is P, X6 is S, X 11 αMeL and X 13 V is X 15 is T, and X 16 Aib and X 17 αMeL and X 19 Q is X 20 is K, and X 21 is I or L, and X 22 L is X 26 R is X 27 A is X 30 A is X 31 R is X 35 is K, and X 37 A is X 38 This is A. In such embodiments, the polypeptide comprises a 42-amino acid sequence.

[0127] Therefore, in some embodiments, this specification refers to polypeptides of formula Ia, or pharmaceutically acceptable salts thereof, GSPSIVTS αMeL DVPT Aib αMeL LQKX 21 Contains LEQERAEKARQQAKEAAEILAQV (Sequence ID 69), X 21 However, if I or L and [X1~X2 does not exist, X 21It is ranked 19th among polypeptides. A polypeptide, or a pharmaceutically acceptable salt thereof, is provided, in which the N-terminal amino acid is optionally N-acetylated and the C-terminal amino acid is optionally amidated.

[0128] In some embodiments of formula Ia, X 21 I is. In some embodiments, the polypeptide includes SEQ ID NO: 20. In other embodiments, the polypeptide is SEQ ID NO: 20. Alternatively, the polypeptide may have at least about 90 to about 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. An exemplary embodiment including SEQ ID NO: 20 is shown. [ka]

[0129] In certain embodiments of formula Ia, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 21 is I, and the polypeptide further comprises a linker-fatty acid. In certain embodiments, the linker-fatty acid is γE-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-εK-CO-(CH2) 18 -CO2H. In certain embodiments of formula Ia, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 35 In this embodiment, K conjugates with a 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 90–97% 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. [ka]

[0130] In certain embodiments of formula Ia, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 21 L is the polypeptide, and the polypeptide further comprises a linker-fatty acid. In certain embodiments, the linker-fatty acid is γE-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-εK-CO-(CH2) 18 -CO2H. In certain embodiments of formula Ia, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 35 In this embodiment, K conjugates with a linker-fatty acid. In another embodiment, the polypeptide comprises SEQ ID NO: 2. Alternatively, the polypeptide may have at least about 90–97% 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. [ka]

[0131] In certain embodiments of formula Ia, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 21 is I, and the polypeptide further comprises a linker-fatty acid. In certain embodiments, the linker-fatty acid is 2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-PKE-γE-CO-(CH2) 18 -CO2H. In certain embodiments of formula Ia, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 35In this embodiment, K conjugates with a linker-fatty acid. In another embodiment, the polypeptide comprises SEQ ID NO: 3. Alternatively, the polypeptide may have at least about 90–97% 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. [ka]

[0132] In certain embodiments of formula Ia, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 21 is I, and the polypeptide further comprises a linker-fatty acid. In certain embodiments, the linker-fatty acid is Ahx-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-γE-CO-(CH2) 18 -CO2H. In certain embodiments of formula Ia, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 35 In this embodiment, K conjugates with a linker-fatty acid. In another embodiment, the polypeptide comprises SEQ ID NO: 4. Alternatively, the polypeptide may have at least about 90–97% 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. [ka]

[0133] In certain embodiments of formula Ia, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 21is I, and the polypeptide further comprises a linker-fatty acid. In certain embodiments, the linker-fatty acid is γE-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-Ahx-CO-(CH2) 18 -CO2H. In certain embodiments of formula Ia, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 35 In this embodiment, K conjugates with a linker-fatty acid. In another embodiment, the polypeptide comprises SEQ ID NO: 5. Alternatively, the polypeptide may have at least about 90–97% 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. [ka]

[0134] In certain embodiments of formula Ia, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 21 is I, and the polypeptide further comprises a linker-fatty acid. In certain embodiments, the linker-fatty acid is γE-GKEKEKE-CO-(CH2) 18 -CO2H. In certain embodiments of formula Ia, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 35 In this embodiment, K conjugates with a linker-fatty acid. In another embodiment, the polypeptide comprises SEQ ID NO: 6. Alternatively, the polypeptide may have at least about 90–97% 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. [ka]

[0135] In certain embodiments of formula Ia, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 21 is I, and the polypeptide further comprises a linker-fatty acid. In certain embodiments, the linker-fatty acid is G-(βA)3-γE-CO-(CH2) 18 -CO2H. In certain embodiments of formula Ia, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 35 In this embodiment, K conjugates with a linker-fatty acid. In another embodiment, the polypeptide comprises SEQ ID NO: 7. Alternatively, the polypeptide may have at least about 90–97% 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. [ka]

[0136] In certain embodiments of formula Ia, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 21 is I, and the polypeptide further comprises a linker-fatty acid. In certain embodiments, the linker-fatty acid is G-(βA)4-γE-CO-(CH2) 18 -CO2H. In certain embodiments of formula Ia, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 35 In this embodiment, K conjugates with a linker-fatty acid. In another embodiment, the polypeptide comprises SEQ ID NO: 8. Alternatively, the polypeptide may have at least about 90–97% 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. [ka]

[0137] In certain embodiments of formula Ia, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 21 is I, and the polypeptide further comprises a linker-fatty acid. In certain embodiments, the linker-fatty acid is 2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-KEKEKE-γE-CO-(CH2) 18 -CO2H. In certain embodiments of formula Ia, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 35 In this embodiment, K conjugates with a linker-fatty acid. In another embodiment, the polypeptide comprises SEQ ID NO: 9. Alternatively, the polypeptide may have at least about 90–97% 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. [ka]

[0138] In certain embodiments of formula Ia, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 21 is I, and the polypeptide further comprises a linker-fatty acid. In certain embodiments, the linker-fatty acid is βA-S-βA-S-γE-CO-(CH2) 18 -CO2H. In certain embodiments of formula Ia, which include polypeptides or pharmaceutically acceptable salts thereof, X 35In this embodiment, K conjugates with a 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 90–97% 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. [ka]

[0139] In certain embodiments of formula Ia, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 21 is I, and the polypeptide further comprises a linker-fatty acid. In certain embodiments, the linker-fatty acid is γE-PKE-Ahx-CO-(CH2) 18 -CO2H. In certain embodiments of formula Ia, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 35 In this embodiment, K conjugates with a 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 90–97% 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. [ka]

[0140] In certain embodiments of formula Ia, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 21L is the polypeptide, and the polypeptide further comprises a linker-fatty acid. In certain embodiments, the linker-fatty acid is γE-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-εK-CO-(CH2) 16 -CO2H. In certain embodiments of formula Ia, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 35 In this embodiment, K conjugates with a 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 90–97% 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. [ka]

[0141] In certain embodiments of formula Ia, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 21 is I, and the polypeptide further comprises a linker-fatty acid. In certain embodiments, the linker-fatty acid is γE-PKE-εK-CO-(CH2) 18 -CO2H. In certain embodiments of formula Ia, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 35 In this case, K conjugates with a linker-fatty acid. Alternatively, the polypeptide may have at least about 90-97% 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. Exemplary embodiments including SEQ ID NO: 13 are shown. [ka]

[0142] In certain embodiments of formula Ia, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 21 is I, and the polypeptide further comprises a linker-fatty acid. In certain embodiments, the linker-fatty acid is γE-PPP-εK-CO-(CH2) 18 -CO2H. In certain embodiments of formula Ia, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 35 In this embodiment, K conjugates with a linker-fatty acid. In another embodiment, the polypeptide comprises SEQ ID NO: 14. Alternatively, the polypeptide may have at least about 90–97% 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. [ka]

[0143] In certain embodiments of formula Ia, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 21 is I, and the polypeptide further comprises a linker-fatty acid. In certain embodiments, the linker-fatty acid is γE-PPPPPP-CO-(CH2) 18 -CO2H. In certain embodiments of formula Ia, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 35 In this embodiment, K conjugates with a linker-fatty acid. In another embodiment, the polypeptide comprises SEQ ID NO: 15. Alternatively, the polypeptide may have at least about 90–97% 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. [ka]

[0144] In certain embodiments of formula Ia, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 21 is I, and the polypeptide further comprises a linker-fatty acid. In certain embodiments, the linker-fatty acid is γE-PEPEPE-CO-(CH2) 18 -CO2H. In certain embodiments of formula Ia, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 35 In this embodiment, K conjugates with a linker-fatty acid. In another embodiment, the polypeptide comprises SEQ ID NO: 16. Alternatively, the polypeptide may have at least about 90–97% 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. [ka]

[0145] In certain embodiments of formula Ia, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 21 is I, and the polypeptide further comprises a linker-fatty acid. In certain embodiments, the linker-fatty acid is γE-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)2-εK-CO-(CH2) 18 -CO2H. In certain embodiments of formula Ia, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 35In this embodiment, K conjugates with a linker-fatty acid. In another embodiment, the polypeptide comprises SEQ ID NO: 17. Alternatively, the polypeptide may have at least about 90–97% 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. [ka]

[0146] In certain embodiments of formula Ia, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 21 is I, and the polypeptide further comprises a linker-fatty acid. In certain embodiments, the linker-fatty acid is γE-εK-γE-εK-CO-(CH2) 18 -CO2H. In certain embodiments of formula Ia, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 35 In this case, K conjugates with a linker-fatty acid. Alternatively, the polypeptide may have at least about 90-99% 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. Exemplary embodiments including SEQ ID NO: 18 are shown. [ka]

[0147] In certain embodiments of formula Ia, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 21 is I, and the polypeptide further comprises a linker-fatty acid. In certain embodiments, the linker-fatty acid is γE-εK-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-εK-CO-(CH2) 18-CO2H. In certain embodiments of formula Ia, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 35 In this embodiment, K conjugates with a 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 90-97% 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. [ka]

[0148] In certain embodiments of formula Ia, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 21 is I, and the polypeptide further comprises a linker-fatty acid. In certain embodiments, the linker-fatty acid is γE-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-εK-CO-(CH2) 16 -CO2H. In certain embodiments of formula Ia, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 35 In this embodiment, K conjugates with a linker-fatty acid. In another embodiment, the polypeptide comprises SEQ ID NO: 21. Alternatively, the polypeptide may have at least about 90–97% 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. [ka]

[0149] In another embodiment of formula I, X1 is Pyr, X2 is G, X3 is S, X4 is P, X5 is S, X6 is G, X 11 αMeL and X 13 V is X 15 is T, and X 16 Aib and X 17 αMeL and X 19 Q is X 20 is K, and X 21 is I and X 22 L is X 26 R is X 27 A is X 30 A is X 31 R is X 35 is K, and X 37 A is X 38 This is A. In such embodiments, the polypeptide comprises a 44-amino acid sequence.

[0150] Therefore, in some embodiments, the present disclosure relates to a polypeptide of formula 1b, or a pharmaceutically acceptable salt thereof. Pyr GSPSGIVTS αMeL DVPT Aib αMeL LQKILEQERAEKARQQAKEAAEILAQV (SEQ ID NO: 70), or a pharmaceutically acceptable salt thereof, The present invention provides polypeptides, or pharmaceutically acceptable salts thereof, in which the C-terminal amino acid is optionally amidated.

[0151] In another embodiment, the polypeptide comprises SEQ ID NO: 23. Alternatively, the polypeptide may have at least about 90–97% 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. An exemplary embodiment comprising SEQ ID NO: 23 is shown. [ka]

[0152] In certain embodiments of formula Ib, comprising a polypeptide or a pharmaceutically acceptable salt thereof, the polypeptide further comprises a linker-fatty acid. In certain embodiments, the linker-fatty acid is γE-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-εK-CO-(CH2) 18 -CO2H. In certain embodiments of formula Ib, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 35 In this embodiment, K conjugates with a linker-fatty acid. In another embodiment, the polypeptide comprises SEQ ID NO: 22. Alternatively, the polypeptide may have at least about 90–97% 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. [ka]

[0153] In certain embodiments of formula Ib, comprising a polypeptide or a pharmaceutically acceptable salt thereof, the polypeptide further comprises a linker-fatty acid. In certain embodiments, the linker-fatty acid is γE-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-εK-CO-(CH2) 16 -CO2H. In certain embodiments of formula Ib, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 35In this embodiment, K conjugates with a linker-fatty acid. In another embodiment, the polypeptide comprises SEQ ID NO: 24. Alternatively, the polypeptide may have at least about 90–97% 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. [ka]

[0154] In another embodiment of formula I, X1 is γE, E, or absent, X2 is absent, X3 is G, X4 is S, X5 is P, X6 is S, X 11 αMeL and X 13 V is X 15 is T, and X 16 Aib and X 17 αMeL and X 19 is Q or E, and X 20 is K, and X 21 is I and X 22 L is X 26 R is X 27 A is X 30 A is X 31 R is X 35 is T, and X 37 A is X 38 A is the answer.

[0155] In another embodiment of formula I, X1 is absent, X2 is absent, X3 is G, X4 is S, X5 is P, X6 is S, X 11 αMeL and X 13 V is X 15 is T, and X 16 Aib and X 17 αMeL and X 19 Q is X20 is K, and X 21 is I and X 22 L is X 26 R is X 27 A is X 30 A is X 31 R is X 35 is T, and X 37 A is X 38 This is A. In such embodiments, the polypeptide comprises a 42-amino acid sequence.

[0156] In another embodiment, the polypeptide comprises SEQ ID NO: 27. Alternatively, the polypeptide may have at least about 90–97% 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. An exemplary embodiment comprising SEQ ID NO: 27 is shown. [ka]

[0157] 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, X 11 αMeL and X 13 V is X 15 is T, and X 16 Aib and X 17 αMeL and X 19 is Q or E, and X 20 is K, and X 21 is I and X 22 L is X 26 R is X 27 A is X 30 A is X 31 R is X 35 is T, and X 37 A is X 38This is A. In such embodiments, the polypeptide comprises a 43-amino acid sequence.

[0158] In certain embodiments of formula Ic, comprising a polypeptide or a pharmaceutically acceptable salt thereof, X1 is γE, and the polypeptide further comprises a linker-fatty acid. In certain embodiments, the polypeptide further comprises a fatty acid conjugated with γE at the X1 position via an amide bond between the alpha-amino group of γE and the carboxylic acid of the linker-fatty acid (2-{2-(2-amino-ethoxy)-ethoxy]acetic acid monomer.

[0159] Therefore, in some embodiments, the present disclosure relates to a polypeptide of formula Ic, or a pharmaceutically acceptable salt thereof. X1GSPSIVTS αMeL DVPT Aib αMeLLX 19 Includes KILEQERAEKARQQATEAAEILAQV (Sequence ID 71), X1 is γE or E, X 19 However, if Q or E and [X2 does not exist, X 19 It is ranked 18th among polypeptides. The present invention provides polypeptides, or pharmaceutically acceptable salts thereof, in which the C-terminal amino acid is optionally amidated.

[0160] In certain embodiments of a polypeptide where X1 is γE, or a pharmaceutically acceptable salt thereof, γE is conjugated with G in X3 via an amide bond between the gamma-carboxylic acid group of γE and the amino group of G.

[0161] In certain embodiments of formula Ic, which comprises a polypeptide or a pharmaceutically acceptable salt thereof, X1 is γE, and X 19 Q is the linker-fatty acid, 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 90-97% 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. An exemplary embodiment comprising SEQ ID NO: 25 is shown. [ka]

[0162] In certain embodiments of formula Ic, which comprises a polypeptide or a pharmaceutically acceptable salt thereof, X1 is γE, and X 19 Q is the linker-fatty acid, 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 90-97% 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. An exemplary embodiment comprising SEQ ID NO: 26 is shown. [ka]

[0163] In certain embodiments of formula Ic, which comprises a polypeptide or a pharmaceutically acceptable salt thereof, X1 is γE, and X 19 Q is the linker-fatty acid, 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 90-97% 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. An exemplary embodiment comprising SEQ ID NO: 28 is shown. [ka]

[0164] In certain embodiments of formula Ic, which comprises a polypeptide or a pharmaceutically acceptable salt thereof, X1 is γE, and X 19 E is the linker-fatty acid, (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 90-97% 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. An exemplary embodiment comprising SEQ ID NO: 52 is shown. [ka]

[0165] In another embodiment of formula 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, X 11 αMeL and X 13 It is D-Val, and X 15 is T or I, and X 16 is Aib or G, and X 17 αMeL and X 19 Q is X 20 is I and X21 is K, and X 22 L is X 26 R is X 27 A is X 30 A is X 31 R is X 35 is K, and X 37 N is X 38 This is T. In such embodiments, the polypeptide comprises a 38-amino acid sequence.

[0166] In some embodiments, the present disclosure relates to a polypeptide of formula Id, or a pharmaceutically acceptable salt thereof. IVTS αMeL D D-Val PX 15 X 16 Contains αMeL LQIKLEQERAEKARQQAKENTEILAQV (Sequence ID 72), X 15 However, if T or I and [X1~X6 does not exist, X 15 It is ranked 9th among polypeptides. X 16 However, if G or Aib and [X1~X6 does not exist, X 16 It is ranked 10th among polypeptides. A polypeptide or a pharmaceutically acceptable salt thereof is provided, wherein the N-terminus is an amino acid, optionally N-acetylated, and the C-terminal amino acid is optionally amidated.

[0167] In certain embodiments of formula Id, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 15 is T, and X 16 The polypeptide is G, and further comprises a linker-fatty acid. In certain embodiments, the linker-fatty acid is 2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)2-γE-CO-(CH2) 18 -CO2H. In certain embodiments of formula Id, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 35In this embodiment, K conjugates with a linker-fatty acid. In another embodiment, the polypeptide is SEQ ID NO: 29. Alternatively, the polypeptide may have at least about 90-97% 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 including SEQ ID NO: 29 is shown. [ka]

[0168] In certain embodiments of formula Id, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 15 is T, and X 16 The polypeptide is G, and further comprises a linker-fatty acid. In certain embodiments, the linker-fatty acid is γE-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-γE-CO-(CH2) 18 -CO2H. In certain embodiments of formula Id, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 35 In this embodiment, K conjugates with a linker-fatty acid. In another embodiment, the polypeptide comprises SEQ ID NO: 30. Alternatively, the polypeptide may have at least about 90–97% 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. [ka]

[0169] In certain embodiments of formula Id, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 15 is T, and X 16The polypeptide is G, and further comprises a linker-fatty acid. In certain embodiments, the linker-fatty acid is γE2-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-CO-(CH2) 18 -CO2H. In certain embodiments of formula Id, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 35 In this embodiment, K conjugates with a 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 90-97% 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. [ka]

[0170] In certain embodiments of formula Id, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 15 is T, and X 16 The polypeptide is G, and further comprises a linker-fatty acid. In certain embodiments, the linker-fatty acid is 2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-εK-γE-CO-(CH2) 18 -CO2. In certain embodiments of formula Id, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 35 In this embodiment, K conjugates with a linker-fatty acid. In another embodiment, the polypeptide comprises SEQ ID NO: 32. Alternatively, the polypeptide may have at least about 90–97% 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. [ka]

[0171] In certain embodiments of formula Id, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 15 is T, and X 16 is G, and the polypeptide further comprises a linker-fatty acid. In certain embodiments, the linker-fatty acid is 2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-γE-εK-CO-(CH2) 18 -CO2H. In certain embodiments of formula Id, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 35 In this embodiment, K conjugates with a linker-fatty acid. In another embodiment, the polypeptide comprises SEQ ID NO: 33. Alternatively, the polypeptide may have at least about 90–97% 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. [ka]

[0172] In certain embodiments of formula Id, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 15 is T, and X 16 The polypeptide is G, and further comprises a linker-fatty acid. In certain embodiments, the linker-fatty acid is εK-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-γE-CO-(CH2) 18 -CO2H. In certain embodiments of polypeptides or pharmaceutically acceptable salts thereof, X 35In this embodiment, K conjugates with a linker-fatty acid. In another embodiment, the polypeptide comprises SEQ ID NO: 34. Alternatively, the polypeptide may have at least about 90–97% 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. [ka]

[0173] In certain embodiments of formula Id, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 15 is T, and X 16 The polypeptide is G, and further comprises a linker-fatty acid. In certain embodiments, the linker-fatty acid is εK-γE-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-CO-(CH2) 18 -CO2H. In certain embodiments of formula Id, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 35 In this embodiment, K conjugates with a linker-fatty acid. In another embodiment, the polypeptide comprises SEQ ID NO: 35. Alternatively, the polypeptide may have at least about 90–97% 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. [ka]

[0174] In certain embodiments of formula Id, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 15 is T, and X 16The polypeptide is G, and further comprises a linker-fatty acid. In certain embodiments, the linker-fatty acid is γE-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-εK-CO-(CH2) 18 -CO2H. In certain embodiments of formula Id, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 35 In this embodiment, K conjugates with a linker-fatty acid. In another embodiment, the polypeptide comprises SEQ ID NO: 36. Alternatively, the polypeptide may have at least about 90–97% 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. [ka]

[0175] In certain embodiments of formula Id, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 15 is T, and X 16 The polypeptide is G, and further comprises a linker-fatty acid. In certain embodiments, the linker-fatty acid is γE-εK-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-CO-(CH2) 18 -CO2H. In certain embodiments of formula Id, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 35 In this embodiment, K conjugates with a linker-fatty acid. In another embodiment, the polypeptide comprises SEQ ID NO: 37. Alternatively, the polypeptide may have at least about 90–97% 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. [ka]

[0176] In certain embodiments of formula Id, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 15 is T, and X 16 is G, and the polypeptide further comprises a linker-fatty acid. In certain embodiments, the linker-fatty acid is γE-γE-εK-CO-(CH2) 18 -CO2H. In certain embodiments of formula Id, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 35 In this embodiment, K conjugates with a linker-fatty acid. In another embodiment, the polypeptide comprises SEQ ID NO: 38. Alternatively, the polypeptide may have at least about 90–97% 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. [ka]

[0177] In certain embodiments of formula Id, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 15 is T, and X 16 is G, and the polypeptide further comprises a linker-fatty acid. In certain embodiments, the linker-fatty acid is γE-εK-γE-CO-(CH2) 18 -CO2H. In certain embodiments of formula Id, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 35In this embodiment, K conjugates with a 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 90-97% 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. [ka]

[0178] In certain embodiments of formula Id, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 15 is T, and X 16 is G, and the polypeptide further comprises a linker-fatty acid. In certain embodiments, the linker-fatty acid is εK-γE-γE-CO-(CH2) 18 -CO2H. In certain embodiments of formula Id, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 35 In this embodiment, K conjugates with a linker-fatty acid. In another embodiment, the polypeptide comprises SEQ ID NO: 40. Alternatively, the polypeptide may have at least about 90–97% 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. [ka]

[0179] In certain embodiments of formula Id, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 15 is T, and X 16Aib is the polypeptide, and the polypeptide further comprises a linker-fatty acid. In certain embodiments, the linker-fatty acid is 2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)2-γE-CO-(CH2) 18 -CO2H. In certain embodiments of formula Id, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 35 In this embodiment, K conjugates with a linker-fatty acid. In another embodiment, the polypeptide comprises SEQ ID NO: 41. Alternatively, the polypeptide may have at least about 90–97% 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. [ka]

[0180] In certain embodiments of formula Id, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 15 is I and X 16 Aib is the polypeptide, and the polypeptide further comprises a linker-fatty acid moiety. In certain embodiments, the linker-fatty acid is 2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)2-γE-CO-(CH2) 18 -CO2H. In certain embodiments of formula Id, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 35 In this embodiment, K conjugates with a linker-fatty acid. In another embodiment, the polypeptide comprises SEQ ID NO: 42. Alternatively, the polypeptide may have at least about 90–97% 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. [ka]

[0181] In certain embodiments of formula Id, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 15 is T, and X 16 is G, and the polypeptide further comprises a linker-fatty acid. In certain embodiments, the linker-fatty acid is GKEKEKE-CO-(CH2) 18 -CO2H. In certain embodiments of formula Id, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 35 In this embodiment, K conjugates with a linker-fatty acid. In another embodiment, the polypeptide comprises SEQ ID NO: 43. Alternatively, the polypeptide may have at least about 90-97% 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. [ka]

[0182] In certain embodiments of formula Id, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 15 is T, and X 16 The polypeptide is G, and further comprises a linker-fatty acid. In certain embodiments, the linker-fatty acid is 2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-γE 2- CO-(CH2) 18 -CO2H. In certain embodiments of formula Id, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 35In this embodiment, K conjugates with a linker-fatty acid. In another embodiment, the polypeptide comprises SEQ ID NO: 44. Alternatively, the polypeptide may have at least about 90–97% 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. [ka]

[0183] In certain embodiments of formula Id, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 15 is I and X 16 is Aib, and the polypeptide further comprises a linker-fatty acid. In certain embodiments, the linker-fatty acid is γE2-CO-(CH2) 18 It is -CO2H). In certain embodiments of formula Id, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 35 In this embodiment, K conjugates with a linker-fatty acid. In another embodiment, the polypeptide comprises SEQ ID NO: 51. Alternatively, the polypeptide may have at least about 90-97% 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. [ka]

[0184] In another embodiment of Equation I, X1 does not exist, X2 does not exist, X3 does not exist, X4 is P, X5 is P, X6 is P, X 11 αMeL and X 13 is V or D-Val, and X15 is T, and X 16 Aib and X 17 αMeL and X 19 Q is X 20 is K, and X 21 is L or Aib, and X 22 L is X 26 R is X 27 A is X 30 A is X 31 R is X 35 is K, and X 37 is A or N, and X 38 is A or T. In such embodiments, the polypeptide comprises a 41-amino acid sequence.

[0185] In some embodiments of this disclosure, the polypeptide, or a pharmaceutically acceptable salt thereof, comprises formula Ie, PPPIVTS αMeL DX 13 PTAib αMeL LQKX 21 LEQERAEKARQQAKEX 37 X 38 Includes EILAQV (Sequence ID 73), X 13 However, if V or D-Val and [X1~X3 does not exist, X 13 It is ranked 10th among polypeptides. X 21 However, if L or Aib and [X1~X3 does not exist, X 21 It is ranked 18th among polypeptides. X 37 However, if A or N and [X1~X3 does not exist, then X 37 It is ranked 34th among polypeptides. X 38 However, if A or T and [X1~X3 does not exist, X 38 It is ranked 35th among polypeptides. The N-terminal amino acid is optionally N-acetylated, and the C-terminal amino acid is optionally amidated.

[0186] In certain embodiments of formula Ie, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 13 It is D-Val, and X 21 Aib and X 37 N is X 38 is T, and the polypeptide further comprises a linker-fatty acid. In certain embodiments, the linker-fatty acid is γE-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-εK-CO-(CH2) 18 -CO2H. In certain embodiments, X 35 In this example, K conjugates with a linker-fatty acid. In some embodiments, the polypeptide comprises SEQ ID NO: 46. Alternatively, the polypeptide may have at least about 90-97% 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. [ka]

[0187] In certain embodiments of formula Id, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 13 It is D-Val, and X 21 L is X 37 A is X 38 A is the polypeptide, and the polypeptide further comprises a linker-fatty acid. In certain embodiments, the linker-fatty acid is γE-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-εK-CO-(CH2) 18 -CO2H. In certain embodiments of formula Id, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 35In this example, K conjugates with a linker-fatty acid. In some embodiments, the polypeptide includes SEQ ID NO: 47. Alternatively, the polypeptide may have at least about 90-97% 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 including SEQ ID NO: 47 is shown. [ka]

[0188] In certain embodiments of formula Id, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 13 V is X 21 L is X 37 A is X 38 A is the polypeptide, and the polypeptide further comprises a linker-fatty acid. In certain embodiments, the linker-fatty acid is (γE-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-εK-CO-(CH2) 18 -CO2H. In certain embodiments of formula Id, which include a polypeptide or a pharmaceutically acceptable salt thereof, X 35 In this example, K conjugates with a linker-fatty acid. In some embodiments, the polypeptide includes SEQ ID NO: 48. Alternatively, the polypeptide may have at least about 90-97% 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 including SEQ ID NO: 48 is shown. [ka]

[0189] In another embodiment of formula I, X1 is γE, X2 is absent, X3 is absent, X4 is G, X5 is P, X6 is S, X 11 αMeL and X 13 It is D-Val, and X 15 is I and X 16 Aib and X 17 αMeL and X 19 Q is X 20 is I and X 21 is K, and X 22 L is X 26 R is X 27 A is X 30 A is X 31 R is X 35 is K, and X 37 N is X 38 is T. In such embodiments, the polypeptide comprises a 42-amino acid sequence. In certain embodiments, the polypeptide further comprises a linker-fatty acid. In certain embodiments, the linker-fatty acid is (2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)2-γE-CO-(CH2) 18 -CO2H. In certain embodiments, which include polypeptides or pharmaceutically acceptable salts thereof, X 35 In this case, K conjugates with linker-fatty acids.

[0190] In some embodiments, the polypeptide includes SEQ ID NO: 45. Alternatively, the polypeptide may have at least about 90-97% 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. Exemplary embodiments including SEQ ID NO: 45 are shown. [ka]

[0191] In another embodiment of formula 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, X 11 L is X 13 V is X 15 is I and X 16 G is X 17 αMeL and X 19 Q is X 20 is K, and X 21 L is X 22 is I and X 26 is K, and X 27 Q is X 30 E is X 31 is K, and X 35 is K, and X 37 N is X 38 This is T. In such embodiments, the polypeptide comprises a 38-amino acid sequence.

[0192] In certain embodiments of the polypeptide, or a pharmaceutically acceptable salt thereof, the polypeptide further comprises a linker-fatty acid. In certain embodiments, the linker-fatty acid is (2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)2γE-C2O-OH). In certain embodiments comprising the polypeptide, or a pharmaceutically acceptable salt thereof, X 35 In this case, K conjugates with linker-fatty acids.

[0193] In some embodiments, the polypeptide includes SEQ ID NO: 49. Alternatively, the polypeptide may have at least about 90-97% 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 including SEQ ID NO: 49 are shown. [ka]

[0194] In another embodiment of formula 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, X 11 L is X 13 V is X 15 is I and X 16 G is X 17 αMeL and X 19 Q is X 20 is K, and X 21 L is X 22 L is X 26 R is X 27 A is X 30 A is X 31 R is X 35 is K, and X 37 A is X 38 This is A. In such embodiments, the polypeptide comprises a 38-amino acid sequence.

[0195] In certain embodiments of the polypeptide, or a pharmaceutically acceptable salt thereof, the polypeptide further comprises a linker-fatty acid. In certain embodiments, the linker-fatty acid is 2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)2-γE-CO-(CH2) 18 -CO2H. In certain embodiments, which include polypeptides or pharmaceutically acceptable salts thereof, X 35 In this case, K conjugates with linker-fatty acids.

[0196] In some embodiments, the polypeptide includes SEQ ID NO: 50. In other embodiments, the polypeptide is SEQ ID NO: 50. Alternatively, the polypeptide may have at least about 90–97% 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. An exemplary embodiment including SEQ ID NO: 50 is shown. [ka]

[0197] In one embodiment, the Specified herein provides a polypeptide or a pharmaceutically acceptable salt thereof comprising a sequence having at least eight amino acid modifications from human urocortin-2 (SEQ ID NO: 53). In some embodiments, the polypeptides provided herein comprise a sequence containing at least one amino acid modification at positions 3, 19, 22, 23, 26, 30, 33, or 37 from human urocortin-2 (SEQ ID NO: 53). In some embodiments, the polypeptides provided herein comprise a sequence containing at least one of the following amino acid modifications from human urocortin-2 (SEQ ID NO: 53): L3T, A19E, R22E, A23K, E26Q, T30E, R33E, R37Q, or any combination thereof.

[0198] In some embodiments, the disclosure provides polypeptides comprising any one of SEQ ID NOs: 1 to 52, or pharmaceutically acceptable salts thereof. Alternatively, the polypeptides may have at least about 90 to 99% sequence similarity to any one of SEQ ID NOs: 1 to 52, or pharmaceutically acceptable salts thereof. Alternatively, the polypeptides may have at least about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence similarity to any one of SEQ ID NOs: 1 to 52.

[0199] In certain embodiments, the polypeptide comprises SEQ ID NO: 1 or a pharmaceutically acceptable salt thereof. In certain embodiments, the polypeptide comprises SEQ ID NO: 2 or a pharmaceutically acceptable salt thereof. In certain embodiments, the polypeptide comprises SEQ ID NO: 22 or a pharmaceutically acceptable salt thereof. In certain embodiments, the polypeptide comprises SEQ ID NO: 25 or a pharmaceutically acceptable salt thereof. In certain embodiments, the polypeptide comprises SEQ ID NO: 29 or a pharmaceutically acceptable salt thereof. In certain embodiments, the polypeptide comprises SEQ ID NO: 41 or a pharmaceutically acceptable salt thereof. In certain embodiments, the polypeptide comprises SEQ ID NO: 42 or a pharmaceutically acceptable salt thereof. In certain embodiments, the polypeptide comprises SEQ ID NO: 45 or a pharmaceutically acceptable salt thereof. In certain embodiments, the polypeptide comprises SEQ ID NO: 46 or a pharmaceutically acceptable salt thereof. In certain embodiments, the polypeptide comprises SEQ ID NO: 48 or a pharmaceutically acceptable salt thereof. In certain embodiments, the polypeptide comprises SEQ ID NO: 49 or a pharmaceutically acceptable salt thereof. In certain embodiments, the polypeptide comprises SEQ ID NO: 50 or a pharmaceutically acceptable salt thereof.

[0200] In certain embodiments, the polypeptide, or a pharmaceutically acceptable salt thereof, is selected from the group consisting of SEQ ID NOs: 1, 2, 22, 25, 29, 41, 42, 45, 46, 48, 49, and 50.

[0201] In certain embodiments of this disclosure, the pharmaceutically acceptable salt is selected from sodium, potassium, trifluoroacetate, hydrochloride, or acetate.

[0202] 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.

[0203] In some embodiments, the present disclosure provides a pharmaceutical composition formulated for oral administration. In other embodiments, the composition is formulated for subcutaneous administration.

[0204] In some embodiments, the present disclosure provides a method for treating a disease or condition selected from the group consisting of diabetes mellitus, 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), comprising administering an effective amount of a polypeptide, or a pharmaceutically acceptable salt thereof, to an individual in need thereof.

[0205] In some embodiments, the present disclosure provides polypeptides or pharmaceutically acceptable salts thereof for use in therapeutic applications.

[0206] In some embodiments, the present disclosure provides polypeptides or pharmaceutically acceptable salts thereof for use in the treatment of diseases or conditions selected from the group consisting of diabetes mellitus, 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).

[0207] In some embodiments, the disclosure provides the use of polypeptides or pharmaceutically acceptable salts thereof in the manufacture of pharmaceuticals for treating diseases or conditions selected from the group consisting of diabetes mellitus, 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).

[0208] In some embodiments, the present disclosure provides a method for treating a disease or condition i selected from the group consisting of diabetes mellitus, 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), comprising administering to a patient an effective amount of polypeptide and an effective amount of additional therapeutic agent.

[0209] In one embodiment, the Specified herein provides polypeptides, or pharmaceutically acceptable salts thereof, that can be used simultaneously with, separately or in succession in combination with, one or more additional therapeutic agents.

[0210] In some embodiments, the present disclosure provides methods for increasing athletic performance or VO2 max.

[0211] In some embodiments, the additional therapeutic agent is a dual agonist of the GIP and GLP-1 receptors. In certain embodiments, the dual agonist of the GIP and GLP-1 receptors is a compound comprising SEQ ID NO: 55.

[0212] In some embodiments, additional therapeutic agents are GIP, GLP-1, and glucagon receptor triagonists. In certain embodiments, the GIP, GLP-1, and glucagon receptor triagonists are compounds comprising SEQ ID NO: 56, SEQ ID NO: 57, or SEQ ID NO: 58, or pharmaceutically acceptable salts thereof.

[0213] In some embodiments, the disclosure provides a method in which the additional therapeutic agent is a glucagon and GLP-1 receptor dual agonist. In certain embodiments, the glucagon and GLP-1 receptor dual agonist 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.

[0214] In some embodiments, the additional therapeutic agent is an amyrin receptor-selective agonist. In certain embodiments, the amyrin receptor-selective agonist is a compound comprising SEQ ID NO: 63, SEQ ID NO: 64, SEQ ID NO: 65, or SEQ ID NO: 66.

[0215] In some embodiments, the additional therapeutic agent is a dual agonist of both calcitonin and amyrin receptors. In certain embodiments, the dual agonist of both calcitonin and amyrin receptors is a compound comprising SEQ ID NO: 67.

[0216] In some embodiments, the additional therapeutic agent is a GLP-1 receptor agonist. In certain embodiments, the GLP-1 receptor agonist is a compound comprising SEQ ID NO: 79, SEQ ID NO: 80, or SEQ ID NO: 81.

[0217] Unless otherwise defined, all technical and scientific terms used herein have the same meanings as those generally understood by those skilled in the art to which this disclosure belongs. Any methods and materials similar or equivalent to those described herein may be used in carrying out or testing polypeptides, pharmaceutical compositions, and methods, but preferred methods and materials are described herein.

[0218] Furthermore, the use of the indefinite article "a" or "an" to refer to an element does not rule out the possibility of multiple elements existing unless the context explicitly requires that there be one or only one element. Therefore, the indefinite article "a" or "an" usually means "at least one."

[0219] As used herein, “about” means within a statistically significant range of a value(s), such as concentration, length, molecular weight, pH, sequence identity, time frame, temperature, or volume. Such a value or range may be within 20%, more typically 10%, and even more typically 5% of a given value or range. The permissible variation encompassed by “about” depends on the particular system in the study and will be readily apparent to those skilled in the art.

[0220] As used herein, “additional therapeutic agent” refers to a substance used to treat, manage, or prevent a disease, other than polypeptides that are active at the CRH2 receptor. In this specification, additional therapeutic agents may include, but are not limited to, dual agonists of the GIP and GLP-1 receptors, triagonists of the GIP, GLP-1, and glucagon receptors, dual agonists of the glucagon and GLP-1 receptors, agonists selective for the amyrin receptor, dual agonists for both the calcitonin and amyrin receptors, agonists of the GLP-1 receptor, or leptin receptor antibodies.

[0221] As used herein, “active,” “activating,” “activating,” etc., with respect to one or more CRH (e.g., CRHR1 or CRHR2) receptors, means the ability of a compound, such as a polypeptide as herein, to bind to and induce a response at the receptor, as measured using assays known in the art, such as the in vitro assays described below.

[0222] As used herein, “amino acids having functional groups available for conjugation” means any natural or non-natural amino acids having functional groups that can be conjugated with fatty acids by a linker, for example. Examples of such functional groups include, but are not limited to, alkynyl, alkenyl, amino, azide, bromo, carboxyl, chloro, iodine, and thiol groups. Examples of natural amino acids containing such functional groups include K (amino), C (thiol), E (carboxyl), and D (carboxyl).

[0223] As used herein, “conservative amino acid substitution” means the substitution of an amino acid with an amino acid that has similar properties (e.g., charge, side chain size, hydrophobic / hydrophilicity, skeletal structure, and rigidity) and has minimal impact on the biological activity of the resulting substituted peptide or polypeptide. Conservative substitutions of functionally similar amino acids are well known in the art and therefore do not need to be described in detail herein.

[0224] As used herein, "at least" means greater than or equal to, or less than or equal to.

[0225] As used herein, body mass index (BMI) refers to a person's weight in kilograms divided by the square of their height in meters.

[0226] As used herein, “cachexia” refers to a complex syndrome associated with an underlying disease that causes progressive muscle loss that does not fully recover with nutritional support. A variety of diseases, including but not limited to cancer, congestive heart failure, chronic obstructive pulmonary disease, chronic kidney disease, and AIDS, can cause cachexia.

[0227] As used herein, “chronic weight management” refers to the long-term management of weight in individuals who are overweight or obese. Chronic weight management may be managed by the polypeptides of this disclosure. In addition to changes in healthy lifestyle (including, but not limited to, a balanced diet and regular physical activity), chronic weight management may also be managed by the polypeptides of this disclosure.

[0228] As used herein, “chronic kidney disease” (CKD) refers to the loss of renal function over time. Persons with CKD may experience one or more of the following signs or symptoms resulting from the loss of renal function: nausea, vomiting, loss of appetite, fatigue, weakness, sleep disturbances, abnormal urination, muscle cramps, swelling of the feet or ankles, hypertension, or shortness of breath.

[0229] As used herein, “conservative amino acid substitution” means the substitution of an amino acid with an amino acid that has similar properties (e.g., charge, side chain size, hydrophobic / hydrophilicity, skeletal structure, and rigidity) and has minimal impact on the biological activity of the resulting substituted peptide or polypeptide. Conservative substitutions of functionally similar amino acids are well known in the art and therefore do not need to be described in detail herein.

[0230] When used herein, "C 16 -C 22 "Fatty acid" refers to a carboxylic acid having 16 to 22 carbon atoms. C is suitable for use in this specification. 16 -C 22 Fatty acids may be saturated monoacids or saturated diacids. As used herein, “saturated” means that the fatty acid does not contain carbon-carbon double or triple bonds.

[0231] As used herein, “diabetes” or “diabetes mellitus” refers to a group of endocrine disorders characterized by high blood glucose levels. Diabetes is used herein to mean either the body’s pancreas failing to produce enough insulin, or the body’s cells becoming unresponsive to hormonal effects. Diabetes is also used herein to mean type 2 diabetes mellitus (T2DM), characterized by high blood glucose, insulin resistance, and / or relative insulin deficiency. As used herein, diabetes also refers to high blood glucose levels that develop in pregnant women, known as gestational diabetes.

[0232] As used herein, “dyslipidemia” refers to abnormally high levels of lipids (fats) in the blood. Dyslipidemia also refers to high levels of cholesterol in the blood.

[0233] As used herein, “effective dose” means the amount, concentration, or dosage of one or more polypeptides described herein, or pharmaceutically acceptable salts thereof, that, after a single or multiple administration to an individual in need, provide the desired effect in such individual under diagnosis or treatment. The effective dose can be readily determined by those skilled in the art by the use of known techniques and by observing results obtained under similar circumstances. In determining the effective dose for a subject, several factors will be considered by the physician, including, but not limited to, the mammalian species; its size, age, and overall health status; the specific disease or disorder involved; the degree or involvement or severity of the disease or disorder; the individual patient’s response; the specific polypeptide administered; the mode of administration; the bioavailability characteristics of the administered preparation; the dose regimen selected; the use of concomitant medications; and other relevant circumstances.

[0234] As used herein, “long-lasting action” means that the binding affinity and activity of the polypeptide persist for a longer period than that of natural human urocortin, allowing for administration at least once daily, or even at a lower frequency of three times, twice, or once per week. The time-acting profile of the polypeptide may be measured using known pharmacokinetic testing methods, such as those used in the following examples.

[0235] As used herein, “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 glucose levels, (2) low HDL cholesterol levels, (3) high blood triglycerides, (4) large waist circumference, and (5) high blood pressure.

[0236] As used herein, “NASH” refers to non-alcoholic steatohepatitis, also known as fatty liver disease. “NASH” also refers to inflammation and damage to the liver caused by the accumulation of fat in the liver. “NASH” also refers to a subtype of non-alcoholic fatty liver disease (“NAFLD”). In some embodiments, “NASH” may be synonymous with “NAFLD”.

[0237] As used herein, the term “obesity” refers to a disorder characterized by excessive body fat that increases the risk of health problems. The term “obesity” also refers to a weight that is heavier than 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 (e.g., hypertension, type 2 diabetes mellitus, or dyslipidemia).

[0238] As used herein, “osteoarthritis” (OA) is a form of arthritis that causes inflammation of one or more joints. “Osteoarthritis” may refer to arthritis affecting the joints of the hands, spine, knees, and hips.

[0239] As used herein, “obstructive sleep apnea” (OSA) refers to sleep-related breathing disorders characterized by recurrent episodes of complete or partial obstruction of the upper airway leading to reduced or absent breathing during sleep.

[0240] As used herein, “patient” and “individual” are used interchangeably and mean mammal, preferably human. In certain embodiments, the patient, preferably human, is further characterized by a disease, disorder, or condition that may benefit from the administration of the polypeptides of this disclosure.

[0241] As used herein, “polypeptide” or “peptide” means a polymer of amino acid residues. This term applies to polymers containing naturally occurring amino acids and polymers containing one or more unnaturally occurring amino acids.

[0242] As used herein, “Polycystic Ovary Syndrome” (PCOS) refers to an endocrine disorder in women of reproductive age. PCOS also refers to a woman who may have one or more of the following conditions: cysts on the ovaries, irregular menstrual periods, heavy periods, excessive hair growth, difficulty conceiving, or insulin resistance.

[0243] As used herein, “individuals in need of it” means mammals such as humans that have a condition, disease, disorder, or symptom in need of treatment or therapy, including, for example, those listed herein.

[0244] As used herein, “quality of weight loss” refers to a reduction in fat mass, but also to the maintenance of lean body mass.

[0245] As used herein, “sarcopenia” refers to a progressive and systemic skeletal muscle disorder characterized by an accelerating loss of muscle mass and function.

[0246] As used herein, “sarcopenic obesity” refers to a person who has both sarcopenia (muscle loss) and obesity. Sarcopenic obesity also refers to the presence of low muscle mass or strength along with high body fat.

[0247] 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.

[0248] % sequence identity = 100% × (number of identical amino acids) / (length of the shortest common sequence) As used herein, the terms "treat", "treatment", "treating" and the like mean inhibition, delay, halting, or reversal of the progression or severity of an existing condition, disease, disorder or symptom.

[0249] Additional non-limiting embodiments are described below.

[0250] Embodiment 1. A polypeptide, or a pharmaceutically acceptable salt thereof, wherein X1X2X3X4X5X6IVTSX 11 DX 13 PX 15 X 16 X 17 LX 19 X 20 X 21 X 22 EQEX 26 X 27 EKX 30 X 31 QQAX 35 EX 37 X 38 comprises EILAQV (SEQ ID NO: 68), X1 is Pyr, γE, or is absent, X2 is G, or is absent, X3 is G, S, or is absent, X4 is S, P, G, or is absent, X5 is S, P, or is absent, X6 is G, S, P, or is absent, X 11 is L or αMeL, X 13 is V or D-Val, X 15 is T or I, X 16 is Aib or G, X 17 is αMeL, X 19 is Q or E, X 20 is K or I, X 21 is I, K, Aib, or L, X 22 is I or L, X 26 is R or K, X 27 is A or Q, X 30 is A or E, X 31 is R or K, X 35 is K or T, X 37 is A or N, X 38 is A or T, a polypeptide, or a pharmaceutically acceptable salt thereof.

[0251] Embodiment 2. GSPSIVTS αMeL DVPT Aib αMeL LQKX 21 LEQERAEKARQQAKEAAEILAQV comprising (SEQ ID NO: 69), X 21 is I or L, the N-terminal amino acid is G at X3, G is N-acetylated, the C-terminal amino acid is V, and V is amidated, the polypeptide according to Embodiment 1, or a pharmaceutically acceptable salt thereof.

[0252] Embodiment 3. X 35 further comprises a linker-fatty acid conjugated to the side chain epsilon-amino group of K at, wherein the K conjugated to the linker-fatty acid is 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)-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-(CH2) 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) 18 -CO2H), 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 A polypeptide according to Embodiment 2, or a pharmaceutically acceptable salt thereof, selected from the group consisting of -CO2H.

[0253] Embodiment 4.X 21However, I is X 35 In this formula, K is represented by K(γE-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-εK-CO-(CH2) 18 A polypeptide according to Embodiment 3, or a pharmaceutically acceptable salt thereof, which conjugates with a fatty acid according to (CO2H).

[0254] Embodiment 5.X 21 However, L is X 35 In this formula, K is represented by K(γE-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-εK-CO-(CH2) 18 A polypeptide according to Embodiment 3, or a pharmaceutically acceptable salt thereof, which conjugates with a fatty acid according to (CO2H).

[0255] Embodiment 6.X 21 However, I is X 35 In this formula, K is K((2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-PKE-γE-CO-(CH2) 18 A polypeptide according to Embodiment 3, or a pharmaceutically acceptable salt thereof, which conjugates with a fatty acid according to (CO2H).

[0256] Embodiment 7.X 21 However, I is X 35 In the formula, K is represented by K(Ahx-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-γE-CO-(CH2) 18 A polypeptide according to Embodiment 3, or a pharmaceutically acceptable salt thereof, which conjugates with a fatty acid according to (CO2H).

[0257] Embodiment 8.X 21 However, I is X 35 In this formula, K is represented by K(γE-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-Ahx-CO-(CH2) 18 A polypeptide according to Embodiment 3, or a pharmaceutically acceptable salt thereof, which conjugates with a fatty acid according to (CO2H).

[0258] Embodiment 9.X 21 is I, and in X 35 K is conjugated to a fatty acid according to the formula K(γE-GKEKEKE-CO-(CH2) 18 -CO2H), the polypeptide according to Embodiment 3, or a pharmaceutically acceptable salt thereof.

[0259] Embodiment 10.X 21 is I, and in X 35 K is conjugated to a fatty acid according to the formula K(G-(βA)3-γE-CO-(CH2) 18 -CO2H), the polypeptide according to Embodiment 3, or a pharmaceutically acceptable salt thereof.

[0260] Embodiment 11.X 21 is I, and in X 35 K is conjugated to a fatty acid according to the formula K(G-(βA)4-γE-CO-(CH2) 18 -CO2H), the polypeptide according to Embodiment 3, or a pharmaceutically acceptable salt thereof.

[0261] Embodiment 12.X 21 is I, and in X 35 K is conjugated to a fatty acid according to the formula K((2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-KEKEKE-γE-CO-(CH2) 18 -CO2H), the polypeptide according to Embodiment 3, or a pharmaceutically acceptable salt thereof.

[0262] Embodiment 13.X 21 is I, and in X 35 K is conjugated to a fatty acid according to the formula K(βA-S-βA-S-γE-CO-(CH2) 18 -CO2H), the polypeptide according to Embodiment 3, or a pharmaceutically acceptable salt thereof.

[0263] Embodiment 14.X 21 is I, and in X 35 K is of the formula K(γE-PKE-Ahx-CO-(CH2)18 A polypeptide according to Embodiment 3, or a pharmaceutically acceptable salt thereof, which conjugates with a fatty acid according to (CO2H).

[0264] Embodiment 15.X 21 However, L is X 35 In this formula, K is represented by K(γE-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-εK-CO-(CH2) 16 A polypeptide according to Embodiment 3, or a pharmaceutically acceptable salt thereof, which conjugates with a fatty acid according to (CO2H).

[0265] Embodiment 16.X 21 However, I is X 35 In this equation, K is given by K(γE-PKE-εK-CO-(CH2) 18 A polypeptide according to Embodiment 3, or a pharmaceutically acceptable salt thereof, which conjugates with a fatty acid according to (CO2H).

[0266] Embodiment 17.X 21 However, I is X 35 In the equation K(γE-PPP-εK-CO-(CH2) 18 A polypeptide according to Embodiment 3, or a pharmaceutically acceptable salt thereof, which conjugates with a fatty acid according to (CO2H).

[0267] Embodiment 18.X 21 However, I is X 35 In the equation K(γE-PPPPPP-CO-(CH2) 18 A polypeptide according to Embodiment 3, or a pharmaceutically acceptable salt thereof, which conjugates with a fatty acid according to (CO2H).

[0268] Embodiment 19.X 21 However, I is X 35 In the equation K(γE-PEPEPE-CO-(CH2) 18 A polypeptide according to Embodiment 3, or a pharmaceutically acceptable salt thereof, which conjugates with a fatty acid according to (CO2H).

[0269] Embodiment 20.X 21 However, I is X 35 In this formula, K is represented by K(γE-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)2-εK-CO-(CH2) 18 A polypeptide according to Embodiment 3, or a pharmaceutically acceptable salt thereof, which conjugates with a fatty acid according to (CO2H).

[0270] Embodiment 21.X 21 However, I is X 35 In the equation K(γE-εK-γE-εK-CO-(CH2) 18 A polypeptide according to Embodiment 3, or a pharmaceutically acceptable salt thereof, which conjugates with a fatty acid according to (CO2H).

[0271] Embodiment 22.X 21 However, I is X 35 In the formula, K is represented by K(γE-εK-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-εK-CO-(CH2) 18 A polypeptide according to Embodiment 3, or a pharmaceutically acceptable salt thereof, which conjugates with a fatty acid according to (CO2H).

[0272] Embodiment 23.X 21 However, the polypeptide described in Embodiment 2, or a pharmaceutically acceptable salt thereof, is I.

[0273] Embodiment 24.X 21 However, I is X 35 In this formula, K is represented by K(γE-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-εK-CO-(CH2) 16 A polypeptide according to Embodiment 3, or a pharmaceutically acceptable salt thereof, which conjugates with a fatty acid according to (CO2H).

[0274] Embodiment 25. The polypeptide according to Embodiment 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 amidated.

[0275] Embodiment 26.X 35 It further contains a linker-fatty acid conjugated with the epsilon-amino group of the side chain at K, and the K conjugated with the linker-fatty acid is K(γE-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-εK-CO-(CH2) 18 -CO2H) and K(γE-(2-[2-(2-aminoethoxy)-ethoxy]-acetyl)-εK-CO-(CH2) 16 A polypeptide according to Embodiment 25, or a pharmaceutically acceptable salt thereof, selected from the group consisting of -CO2H).

[0276] Embodiment 27.X 35 In this formula, K is represented by K(γE-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-εK-CO-(CH2) 18 The polypeptide according to Embodiment 26, which conjugates with a fatty acid according to -CO2H.

[0277] Embodiment 28.X 35 In this formula, K is represented by K(γE-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-εK-CO-(CH2) 16 The polypeptide according to Embodiment 26, which conjugates with a fatty acid according to -CO2H.

[0278] Embodiment 29. X1GSPSIVTS αMeL DVPT Aib αMeL LX 19 Includes KILEQERAEKARQQATEAAEILAQV (Sequence ID 71), X1 is either γE or does not exist. X2 does not exist. X 19However, it is either Q or E, The C-terminal amino acid is X 44 It is a V of position, X 44 The polypeptide according to Embodiment 1, or a pharmaceutically acceptable salt thereof, wherein the V at position is amidated.

[0279] Embodiment 30. The polypeptide according to Embodiment 29, wherein X1 is γE, and γE in X1 is conjugated with G in X3 via an amide bond between the γ-carboxylic acid group of γE and the amino group of G.

[0280] Embodiment 31. Further comprising a fatty acid conjugated with γE at the X1 position via an amide bond between the alpha-amino group of γE and the carboxylic acid group of (2-[2-(2-amino-ethoxy)-ethoxy]acetic acid monomer, wherein the linker-fatty acid is (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 A polypeptide according to embodiment 30, selected from the group consisting of -CO2H.

[0281] In Embodiment 32.X1, the linker-fatty acid conjugated with γE is (2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-εK-CO-(CH2) 18 -CO2H, X 19 However, Q is the polypeptide described in Embodiment 31.

[0282] In Embodiment 33.X1, the linker-fatty acid conjugated with γE is (2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-K-CO-(CH2) 18 -CO2H, X 19However, Q is the polypeptide described in Embodiment 31.

[0283] Embodiment 34.X1 does not exist, X 19 However, Q is the polypeptide described in Embodiment 31.

[0284] In Embodiment 35.X1, the linker-fatty acid conjugated with γE is (2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-εK-CO-(CH2) 16 -CO2H, X 19 However, Q is the polypeptide described in Embodiment 31.

[0285] In Embodiment 36.X1, the linker-fatty acid conjugated with γE is (2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-Dap-CO-(CH2) 18 -CO2H, X 19 However, the polypeptide described in Embodiment 31 is E.

[0286] Embodiment 37. IVTS αMeL D D-Val PX 15 X 16 Contains αMeL LQIKLEQERAEKARQQAKENTEILAQV (SEQ ID NO: 72), or a pharmaceutically acceptable salt thereof. X 15 However, it is T or I, X 16 However, it is G or Aib, The N-terminal amino acid is I at position X7, and the I at position X7 is N-acetylated, and the C-terminal amino acid is X 44 It is a V of position, X 44 The polypeptide according to Embodiment 1, or a pharmaceutically acceptable salt thereof, wherein the V at position is amidated.

[0287] Embodiment 38.X 35It further contains a linker-fatty acid conjugated with the epsilon-amino group of the side chain at K, and the K conjugated with the linker-fatty acid is 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((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-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-CO-(CH2) 18 -CO2H), 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)-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 A polypeptide according to embodiment 37, selected from the group consisting of -CO2H).

[0288] Embodiment 39.X 15 However, T is X 16 However, G is X 35 In this formula, K is represented by K((2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)2-γE-CO-(CH2) 18 The polypeptide according to embodiment 38, which conjugates with a fatty acid according to -CO2H.

[0289] Embodiment 40.X 15 However, T is X 16 However, G is X 35 In this formula, K is represented by K(γE-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-γE-CO-(CH2) 18 The polypeptide according to embodiment 38, which conjugates with a fatty acid according to -CO2H.

[0290] Embodiment 41.X 15 However, T is X 16 However, G is X 35 In this formula, K is represented by K(γE2-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-CO-(CH2) 18 The polypeptide according to embodiment 38, which conjugates with a fatty acid according to -CO2H.

[0291] Embodiment 42.X 15 However, T is X 16 However, G is X 35 In this formula, K is represented by K((2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-εK-γE-CO-(CH2) 18 The polypeptide according to embodiment 38, which conjugates with a fatty acid according to -CO2H.

[0292] Embodiment 43.X 15 However, T is X 16 However, G is X 35 In this formula, K is represented by K((2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-γE-εK-CO-(CH2) 18The polypeptide according to embodiment 38, which conjugates with a fatty acid according to -CO2H.

[0293] Embodiment 44.X 15 However, T is X 16 However, G is X 35 In the formula, K is represented by K(εK-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-γE-CO-(CH2) 18 The polypeptide according to embodiment 38, which conjugates with a fatty acid according to -CO2H.

[0294] Embodiment 45.X 15 However, T is X 16 However, G is X 35 In this formula, K is represented by K(εK-γE-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-CO-(CH2) 18 The polypeptide according to embodiment 38, which conjugates with a fatty acid according to -CO2H.

[0295] Embodiment 46.X 15 However, T is X 16 However, G is X 35 In this formula, K is represented by K(γE-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-εK-CO-(CH2) 18 The polypeptide according to embodiment 38, which conjugates with a fatty acid according to -CO2H.

[0296] Embodiment 47.X 15 However, T is X 16 However, G is X 35 In the formula, K is represented by K(γE-εK-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-CO-(CH2) 18 A polypeptide according to embodiment 38, which conjugates with a fatty acid according to -CO2H.

[0297] Embodiment 48.X 15 However, T is X 16 However, G is X 35In this equation, K is given by K(γE-γE-εK-CO-(CH2) 18 The polypeptide according to embodiment 38, which conjugates with a fatty acid according to -CO2H.

[0298] Embodiment 49.X 15 However, T is X 16 However, G is X 35 In this equation, K is given by K(γE-εK-γE-CO-(CH2) 18 The polypeptide according to embodiment 38, which conjugates with a fatty acid according to -CO2H.

[0299] Embodiment 50.X 15 However, T is X 16 However, G is X 35 In this equation, K is given by K(εK-γE-γE-CO-(CH2) 18 The polypeptide according to embodiment 38, which conjugates with a fatty acid according to -CO2H.

[0300] Embodiment 51.X 15 However, T is X 16 However, Aib and X 35 In this formula, K is represented by K((2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)2-γE-CO-(CH2) 18 The polypeptide according to embodiment 38, which conjugates with a fatty acid according to -CO2H.

[0301] Embodiment 52.X 15 However, I is X 16 However, Aib and X 35 In this formula, K is represented by K((2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)2-γE-CO-(CH2) 18 The polypeptide according to embodiment 38, which conjugates with a fatty acid according to -CO2H.

[0302] Embodiment 53.X 15 However, T is X 16 However, G is X 35 In the equation K(GKEKEKE-CO-(CH2)18 The polypeptide according to embodiment 38, which conjugates with a fatty acid according to -CO2H.

[0303] Embodiment 54.X 15 However, T is X 16 However, G is X 35 In this formula, K is represented by K((2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-γE2-CO-(CH2) 18 The polypeptide according to embodiment 38, which conjugates with a fatty acid according to -CO2H.

[0304] Embodiment 55.X 15 However, I is X 16 However, Aib and X 35 In the equation K(γE2-CO-(CH2) 18 The polypeptide according to embodiment 38, which conjugates with a fatty acid according to -CO2H.

[0305] Embodiment 56. PPPIVTS αMeL DX 13 PTAib αMeL LQKX 21 LEQERAEKARQQAKEX 37 X 38 Includes EILAQV (Sequence ID 73), X 13 However, it is V or D-Val, X 21 However, it is L or Aib, X 37 However, it is either A or N, X 38 However, it is A or T, The N-terminal amino acid is P in X4, and the P in X4 is N-acetylated, and the C-terminal amino acid is X 44 V in X 44 The polypeptide described in Embodiment 1, or a pharmaceutically acceptable salt thereof, wherein V in the polypeptide is amidated.

[0306] Embodiment 57. X1 is γE, X2 does not exist. X3 does not exist. X4 is G, X5 is P, X6 is S, X 11 However, it is αMeL, X 13 However, it is D-Val, X 15 However, it is I, X 16 However, it is Aib, X 19 However, Q is, X 20 However, it is I, X 21 However, K is, X 22 However, it is L, X 26 However, R is, X 27 However, A is, X 30 However, A is, X 31 However, R is, X 35 However, K is, X 37 However, N is, X 38 However, T is, The C-terminal amino acid is X 44 It is a V of position, X 44 The polypeptide according to Embodiment 1, or a pharmaceutically acceptable salt thereof, wherein the V at position is amidated.

[0307] Embodiment 58.X 35 It further contains a linker-fatty acid conjugated with the epsilon-amino group of the side chain at K, and the K conjugated with the linker-fatty acid is K((2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)2-γE-CO-(CH2) 18 -CO2H) and K(γE-(2-[2-(2-aminoethoxy)-ethoxy]-acetyl)-εK-CO-(CH2) 18A polypeptide according to Embodiment 56, or a pharmaceutically acceptable salt thereof, selected from the group consisting of -CO2H.

[0308] Embodiment 59.X 13 However, it is D-Val, and X 21 However, Aib and X 35 In this formula, K is represented by K(γE-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-εK-CO-(CH2) 18 -CO2H) conjugates with fatty acids, X 37 However, N is X 38 However, the polypeptide described in Embodiment 58 is T.

[0309] Embodiment 60.X 13 However, it is D-Val, and X 21 However, L is X 35 In this formula, K is represented by K(γE-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-εK-CO-(CH2) 18 -CO2H) conjugates with fatty acids, X 37 However, A is X 38 However, the polypeptide described in Embodiment 58 is A.

[0310] Embodiment 61.X 13 However, V is X 21 However, L is X 35 In this formula, K is represented by K(γE-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-εK-CO-(CH2) 18 -CO2H) conjugates with fatty acids, X 37 However, A is X 38 However, the polypeptide described in Embodiment 58 is A.

[0311] Embodiment 62. X1 does not exist, X2 does not exist. X3 does not exist. X4 does not exist, X5 does not exist. X6 does not exist. X 11 However, it is L, X 13 However, V is, X 15 However, it is I, X 16 However, it is G, X 19 However, Q is, X 20 However, K is, X 21 However, it is L, X 22 However, it is L, X 26 However, K is, X 27 However, Q is, X 30 However, E is, X 31 , K, X 35 However, K is, X 37 However, N is, X 38 However, T is, The N-terminal amino acid is I in X7, and I in X7 is N-methylated. The C-terminal amino acid is X 44 It is a V of position, X 44 The polypeptide according to Embodiment 1, or a pharmaceutically acceptable salt thereof, wherein the V at position is amidated.

[0312] Embodiment 63.X 35 The formula K((2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)2-γE-C) further contains a linker-fatty acid conjugated with the epsilon-amino group of the side chain at K, and the K conjugated with the linker-fatty acid is given by formula K((2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)2-γE-C) 20 A polypeptide according to Embodiment 62, or a pharmaceutically acceptable salt thereof, which is of the form -OH.

[0313] Embodiment 64. X1 does not exist, X2 does not exist. X3 does not exist. X4 does not exist, X5 does not exist. X6 does not exist. X 11 However, it is L, X 13 However, V is, X 15 However, it is I, X 16 However, it is G, X 19 However, Q is, X 20 However, K is, X 21 However, it is L, X 22 However, it is L, X 26 However, R is, X 27 However, A is, X 30 However, A is, X 31 However, R is, X 35 However, K is, X 37 However, A is, X 38 However, A is, The N-terminal amino acid is I in X7, and I in X7 is N-methylated. The C-terminal amino acid is X 44 It is a V of position, X 44 The polypeptide according to Embodiment 1, or a pharmaceutically acceptable salt thereof, wherein the V at position is amidated.

[0314] Embodiment 65.X 35 The formula further contains a linker-fatty acid conjugated with the epsilon-amino group of the side chain at K, and the K conjugated with the linker-fatty acid is given by formula K((2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)2-γE-CO-(CH2) 18 A polypeptide according to Embodiment 64, or a pharmaceutically acceptable salt thereof, which is of the form -CO2H.

[0315] Embodiment 66. A polypeptide comprising any one of SEQ ID NOs: 1 to 52, or a pharmaceutically acceptable salt thereof.

[0316] Embodiment 67. A polypeptide according to any one of Embodiments 1 to 66, or a pharmaceutically acceptable salt thereof, wherein the pharmaceutically acceptable salt is selected from sodium, potassium, trifluoroacetate, hydrochloride, and acetate.

[0317] Embodiment 68. A pharmaceutical composition comprising a polypeptide according to any one of Embodiments 1 to 66, or a pharmaceutically acceptable salt thereof, and at least one pharmaceutically acceptable carrier, diluent, or excipient.

[0318] Embodiment 69. The pharmaceutical composition according to Embodiment 68, wherein the composition is formulated for oral administration.

[0319] Embodiment 70. The pharmaceutical composition according to Embodiment 68, wherein the composition is formulated for subcutaneous administration.

[0320] Embodiment 71. A method for treating a disease or condition selected from the group consisting of diabetes mellitus, 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), comprising administering an effective amount of a polypeptide described in any one of Embodiments 1 to 66, or a pharmaceutically acceptable salt thereof, to an individual in need thereof.

[0321] Embodiment 72. A polypeptide according to any one of Embodiments 1 to 66, or a pharmaceutically acceptable salt thereof, for use in therapy.

[0322] Embodiment 73. A polypeptide according to any one of Embodiments 1 to 66, or a pharmaceutically acceptable salt thereof, for use in treating a disease or condition selected from the group consisting of diabetes mellitus, 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).

[0323] Embodiment 74. Use of a polypeptide according to any one of Embodiments 1 to 66, or a pharmaceutically acceptable salt thereof, in the manufacture of a pharmaceutical product for treating a disease or condition selected from the group consisting of diabetes mellitus, 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).

[0324] Embodiment 75. A method for treating a disease or condition selected from the group consisting of diabetes mellitus, 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) in a patient in need thereof, comprising administering to the patient an effective amount of a polypeptide described in any one of Embodiments 1 to 66 and an effective amount of an additional therapeutic agent.

[0325] Embodiment 76. The therapeutic method according to Embodiment 75, wherein the additional therapeutic agent is a dual agonist of GIP and GLP-1 receptors.

[0326] Embodiment 77. The method according to Embodiment 76, wherein the dual agonist of GIP and GLP-1 receptors is a compound comprising SEQ ID NO: 55.

[0327] Embodiment 78. The therapeutic method according to Embodiment 75, wherein the additional therapeutic agonist is a triagonist of GIP, GLP-1, and a glucagon receptor.

[0328] Embodiment 79. The method according to Embodiment 78, wherein GIP, GLP-1, and a glucagon receptor triagonist are compounds comprising SEQ ID NO: 56, SEQ ID NO: 57, or SEQ ID NO: 58, or pharmaceutically acceptable salts thereof.

[0329] Embodiment 80. The therapeutic method according to Embodiment 75, wherein the additional therapeutic agent is a dual agonist of glucagon and a GLP-1 receptor.

[0330] Embodiment 81. The method according to Embodiment 80, wherein the glucagon and GLP-1 receptor dual agonist 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.

[0331] Embodiment 82. The therapeutic method according to Embodiment 75, wherein the additional therapeutic agent is an amyrin receptor-selective agonist.

[0332] Embodiment 83. The method according to Embodiment 82, wherein the amyrin receptor-selective agonist is a compound comprising SEQ ID NO: 63, SEQ ID NO: 64, SEQ ID NO: 65, or SEQ ID NO: 66.

[0333] Embodiment 84. The therapeutic method according to Embodiment 75, wherein the additional therapeutic agent is a dual agonist of both calcitonin and amyrin receptors.

[0334] Embodiment 85. The method according to Embodiment 84, wherein the dual agonist for both calcitonin and amyrin receptors is a compound comprising SEQ ID NO: 67.

[0335] Embodiment 86. The therapeutic method according to Embodiment 75, wherein the additional therapeutic agent is a GLP-1 receptor agonist.

[0336] Embodiment 87. The method according to Embodiment 86, wherein the GLP-1 receptor agonist is a compound comprising SEQ ID NO: 79, SEQ ID NO: 80, or SEQ ID NO: 81. [Examples]

[0337] Example 1: Preparation and purification of polypeptides containing SEQ ID NO: 20 and SEQ ID NO: 1 Polypeptides containing SEQ ID NO: 20 and SEQ ID NO: 1 were prepared according to the following steps. First, polypeptides containing SEQ ID NO: 20 were synthesized using the fluorenylmethyloxycarbonyl (Fmoc) / tert-butyl (t-Bu) chemical reaction in a Symphony 12-channel multiplex peptide synthesizer (Protein Technologies, Inc., Tucson, AZ). During solid-phase synthesis, the glass reaction vessel was wrapped in heat tape (BriskHeat controller: Model SDC-120JC-A; tape: Model BS0051080), and the heat tape temperature was set to 60°C for all reactions except the removal of the Mtt protecting group, which proceeded at ambient temperature.

[0338] Polystyrene Rink Amide MBHA resin LL (Novabiochem, sub: 0.39 meq / g, 100-200 mesh, catalog number 855045) was used for synthesis on a 0.13 mmol scale. Unless otherwise specified, standard side-chain protecting groups were used. Fmoc-Lys(Mtt)-OH was used as the lysine to be acylated. Before each coupling step (2 times x 5 minutes), the Fmoc group was removed using 20% ​​piperidine in DMF. All amino acid couplings were performed for 30 minutes at a molar excess of 7.7 times the theoretical peptide load, using equimolar ratios of Fmoc amino acid (0.3 M), diisopropylcarbodiimide (0.9 M), and Oxyma (0.9 M). αMeL and post-αMeL amino acid coupling were performed for 6 hours. After skeleton synthesis, the N-terminal amino acid was deprotected, then mixed with DCM for 30 minutes, and drained. Next, 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 a standard single-letter amino acid code excluding the N-terminal acetyl group, non-coding residues (αMeL and Aib), and the C-terminal valineamide, with these modifications and the structure of the amino acid residues extended. [ka]

[0339] Following N-terminal acetylation, the Mtt protecting group on the lysine acylated peptide was selectively removed from the resin-bound peptide using six treatments (6 × 15 minutes each) of a DCM solution of 30% hexafluoroisopropanol (HFIP; Oakwood Chemical). DMF washing was performed to remove any remaining HFIP before the next step.

[0340] For all acylated polypeptides described herein, subsequent linker-fatty acid moiety bonding is achieved by performing the Fmoc / tBu procedure described above to obtain the desired order and combination of amino acid monomers with the standard protecting group, as well as the following non-standard monomers: N α -Boc-N ε -Fmoc-L-lysine (Boc-L-Lys(Fmoc)-OH, Chem-Impex Int'l Inc), N α -Fmoc-N β -Boc-L-2,3-diaminopyropionic acid (Fmoc-Dap(Boc)-OH, Chem-Impex Int'l Inc.), Fmoc-β-alanine (Fmoc-β-Ala-OH, Orpegen), Fmoc-6-aminohexanoic acid (Fmoc-Ahx-OH, listed as Fmoc-ε-Acp-OH by supplier Chem-Impex Int'l Inc.), Fmoc-L-glutamic acid α-t-butyl ester (Fmoc-L-Glu-OtBu, Ark Pharm, Inc.), 1-(9H-fluoren-9-yl)-3-oxo-2,7,10-trioxa-4-azadodecane-12-acid (Fmoc-AEEA-OH, ChemShuttle), mono-OtBu-octadecanediic acid (WuXi AppTec, Shanghai, China, and mono-OtBu-eicosanedioic acid (WuXi AppTec, Shanghai, China). For SEQ ID NO: 1, the order was Fmoc-Glu-OtBu, Fmoc-AEEA-OH, Boc-L-Lys(Fmoc)-OH, mono-OtBu-eicosanedioic acid. A 3-fold excess of reagent (equomolar AA:DIC:Oxyma) was used for each coupling cycle of 6 hours.

[0341] After synthesis was complete, the peptide resin was washed with DCM and then completely air-dried. The dried resin was treated with 10 mL of a cutting cocktail (trifluoroacetic acid / TFA:1,2-ethanedithiol:triisopropylsilane:water = 89:3:3:5v / v) at room temperature for 2 hours. The resin was collected by filtration, and the filtrate was treated with 4x cold diethyl ether (-20°C) to precipitate the crude peptide. The peptide / ether suspension was then centrifuged at 3200 RCF for 1.5 minutes to form a solid pellet, the supernatant was drained, and the solid pellet was further pulverized twice with ether. The air-dried pellet was dissolved in acetic acid (5 mL) and water (10 mL).

[0342] Crude peptides were purified by RP-HPLC using a linear gradient with 100% acetonitrile and a 0.05% TFA / water buffer system on a Phenomenex PhenylHexyl column (5 μm, 100 A, 250 × 21.2 mm, part number: 00G-4257-P0-AX). Peptide purity was evaluated using analytical RP-HPLC with a Waters Acuity CSH C18 column (1.7 μm, 2.1 × 100 mm, part number: 186005572), and the pool standard was over 90%. The main pool purity of SEQ ID NO: 20 was found to be over 92.0%. Subsequent lyophilization of the final main product pool yielded the lyophilized peptide TFA salt. The molecular weight of SEQ ID NO: 20 was determined by LC / MS (measured value: [M + 3H]). 3+ =1540.1; Calculated value [M+3H] 3+ =1539.1; measured value MW (average) = 4617.3; calculated value MW (average) = 4614.3). The molecular weight of Sequence ID No. 1 was determined by LC / MS (measured value: [M+3H] 3+ =1781.1; Calculated value [M+3H] 3+=1781.4; measured MW (average) = 5340.3; calculated MW (average) = 5341.3). The following is a schematic diagram of the polypeptide containing Sequence ID No. 1, using the standard single-letter amino acid code excluding the N-terminal acetyl group, non-coding residues (αMeL and Aib), acylated lysine at position 35, and the C-terminal valineamide, with these modifications and the structure of the amino acid residues extended. [ka]

[0343] Using processes similar to those described above and those known to those skilled in the art, the peptide skeleton can be synthesized, the fatty acid-linker moiety conjugated, its purity examined, and the molecular weight of the polypeptide of the present invention as described herein can be confirmed. Compounds according to SEQ ID NOs: 2-19, 21, 29-44, 46-48, and 51 were prepared substantially as described in the procedure of Example 1.

[0344] Example 2: Preparation and purification of polypeptides containing SEQ ID NO: 23 and SEQ ID NO: 22 Solid-phase synthesis was carried out in the same manner as in Example 1, except 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) was coupled under the standard conditions described in Example 1.

[0345] Sequence ID 45 was prepared in the same manner as the N-terminal Pyr, but using Boc-Glu-OtBu (Synthonix). Sequence IDs 49 and 50 were prepared in the same manner as the N-terminal Pyr, but using Boc-NMeI (Combi-Blocks).

[0346] The following is a schematic diagram of Sequence ID No. 23, using the standard single-letter amino acid code excluding the N-terminal l-Pyr-OH, non-coding residues (αMeL and Aib), and the C-terminal valineamide, with these modifications and the structure of the amino acid residues extended. [ka]

[0347] The molecular weight of SEQ ID NO: 23 was determined by LC / MS (measured value: [M+3H]). 3+ =1582.1; Calculated value [M+3H] 3+ =1581.2; Measured value MW (average) = 4743.3; Calculated value MW (average) = 4740.5).

[0348] The following is a schematic diagram of Sequence ID No. 22, using the standard single-letter amino acid code excluding the N-terminal l-Pyr-OH, non-coding residues (αMeL and Aib), acylated lysine at position 35, and the C-terminal valineamide, with these modifications and the structure of the amino acid residues extended. [ka]

[0349] Molecular weight is determined by LC / MS (measured value: [M+3H]) 3+ =1823.4; Calculated value [M+3H] 3+ =1823.5; measured MW (average) = 5467.2; calculated MW (average) = 5467.4). The compound according to Sequence ID No. 24 was prepared substantially as described in the procedure of Example 2.

[0350] Example 3: Preparation and purification of polypeptides containing SEQ ID NO: 27 and SEQ ID NO: 25 For this polypeptide and other polypeptides that were ultimately N-acylated with a linker-fatty acid group, N-terminal acetylation was not appropriate, and the synthesis did not include the acetic anhydride step described in Example 1. For SEQ ID NO: 27, the synthesis was terminated by removal of the Fmoc protecting group on the N-terminal glycine residue.

[0351] The following is a schematic diagram of Sequence ID No. 27, using the standard single-letter amino acid code excluding the N-terminal glycine, non-coding residues (αMeL and Aib), and the C-terminal valineamide, with these modifications and the structure of the amino acid residues extended. [ka]

[0352] The molecular weight of SEQ ID NO: 27 was determined by LC / MS (measured value: [M+3H]). 3+ =1516.9; Calculated value [M+3H] 3+ =1516.1; Measured value MW (average) = 4547.7; Calculated value MW (average) = 4545.2).

[0353] For SEQ ID NO: 25 and other polypeptides N-acylated with a linker-fatty acid group (SEQ ID NOs: 26, 28, 52), Fmoc-Lys(Mtt)-OH was not used in the sequence, and the synthesis continued without the 30% HFIP deprotection step described in Example 1. For SEQ ID NO: 25, after the final Fmoc deprotection step of the synthesis of SEQ ID NO: 27, the synthesis was completed using the Fmoc / tBu chemistry as described in Example 1, and the coupling / Fmoc deprotection step was performed for the monomers added in the following order: Fmoc-Glu-OtBu, Fmoc-AEEA-OH, Boc-L-Lys(Fmoc)-OH, mono-OtBu-eicosanedioic acid.

[0354] The following is a schematic diagram of Sequence ID No. 25, using the standard single-letter amino acid code excluding the N-terminal acylated glycine, non-coding residues (αMeL and Aib), and the C-terminal valineamide, with these modifications and the structure of the amino acid residues extended. [ka]

[0355] The molecular weight was determined by LC / MS (measured value: [M+3H]). 3+ =1758.1; Calculated value [M+3H] 3+ =1758.4; measured MW (average) = 5271.3; calculated MW (average) = 5272.2). Compounds according to Sequence IDs 26, 28, and 52 were prepared substantially according to the procedure described in Example 3.

[0356] [Table 1]

[0357] Example 4. In vitro evaluation of CRHR1 and CRHR2 activity via intracellular cAMP production. Corticotropin-releasing hormone (CRH) receptors 1 and 2 (CRHR1 and CRHR2) are G s These are GPCRs functionally coupled to proteins. Stimulation of these receptors increases intracellular cAMP production, which can be detected using standard in vitro techniques. The in vitro activity of the peptides was measured by the amount of cAMP formed in human CRHR1 and CRHR2 overexpressing cells.

[0358] Human CRHR1 and CRHR2 expressing cell lines Human CRHR1 and CRHR2 receptors were stably expressed in human embryonic kidney cells (HEK293) under the control of a pcDNA expression vector. HEK293 cell lines were cultured in 1× DMEM (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 plasmids were transfected into HEK293 cells using Lipo fectAMINE 3000 transfection reagent (Invitrogen, L3000001). After 20 days under selective conditions, mRNA levels from different clones were measured to confirm the expression of human CRHR1 and CRHR2 genes. To determine the function of overexpressed CRHR cells, intracellular cAMP levels in response to human UCN1 and UCN2 were measured and compared with the expression of CRHR1 and CRHR2 in each clone.

[0359] 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 ug / 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 ug / ml puromycin. The cultured cells were grown to a concentration of 90% and harvested. The cells were frozen in a mixture of 100% fetal bovine serum + 7% DMSO at 10 × 10 e6 / ml and stored in liquid nitrogen until use.

[0360] Native cell lines expressing endogenous human CRHR1 and CRHR2 The human retinoblastoma cell line Y79 expresses high levels of human CRHR1 but does not express detectable CRHR2. The rat aortic smooth muscle cell line A7r5 expresses only the CRHR2 receptor without detectable CRHR1. The rat CRHR2 gene shares 93% homology of human CRHR2 nucleotide sequence and responds to human UCN2. Therefore, these two native cell lines were used to further characterize polypeptide selectivity.

[0361] Y79 cells were grown in suspension culture as recommended by ATCC in a medium containing RPMI (Gibco catalog no. 11875093) and 20% FBS (Gibco dialysis, catalog no. 26400044). The Y79 cells were incubated at 37°C and 5% CO2. After centrifugation, the supernatant was removed and the cell pellet was subcultured by resuspending it in fresh medium (the subculture ratio was 1:5). Adherent 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 harvested and frozen at 10 × 10 e6 / mL in a mixture of 100% fetal bovine serum + 7% DMSO, and stored in liquid nitrogen until use.

[0362] Human CRHR1 receptor and CRHR2 cAMP assay On the day of the assay, frozen cells were thawed and diluted with assay buffer (MEM without phenol red (Corning, 17-305-CV, lot number 32121010), 0.1% casein (Sigma C4765, lot number SLCG2120), 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 plate (Corning catalog number 354661). The test compound, diluted with DMSO, was added in a 1:3 dilution series using an ECHO acoustic liquid handler (Beckman) (200 nL / well). The cell and test compound mixtures were incubated at 37°C for 1 hour for the hCRHR1 plates and at room temperature for 30 minutes for the CRHR2 plates.

[0363] The amount of intracellular cAMP was quantified using HTRF (homogenous time-resolved fluorescence) technology (Cisbio) according to the vendor's instructions. Briefly, 10 μL of CAMP-d2 conjugate and 10 μL of anti-CAMP-cryptate conjugate in lysis buffer were incubated with treated cells at room temperature for 60 minutes. The HTRF signal was immediately detected using an Envision plate reader (Perkin-Elmer), and the fluorescence ratio at 665–620 nm was calculated.

[0364] Human Y79 and rat A7r5 cAMP assay The cAMP assay for Y79 and A7r5 cells was performed in the same manner as the hCRHR1 and hCRHR2 assays. Briefly, on the day of the assay, frozen cells were thawed and diluted with assay 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 plate. The test compound, diluted with DMSO, was added in a 1:3 dilution series using an ECHO acoustic liquid handler (Beckman) (200 nL / well). The cell and test compound mixtures were incubated at 37°C for 1 hour for both Y79 and A7r5 cells.

[0365] Data Analysis The raw fluorescence data for the peptide concentration curves in Tables 2 and 3 were converted to cAMP amounts (pmol / well) using the cAMP standard curve generated for each experiment. Using a 4-parameter logistic curve fitting program (Genedata Screener® v12.0.4), relative EC50 values ​​were calculated from the upper and lower ranges of the concentration-response curves defined using 10 nM human UCN2 as the maximum value for CRHR2 cells, and 66 nM human UCN1 and buffer alone as the minimum value for CRHR1 cells.

[0366] result [Table 2]

[0367] This table shows that all polypeptides in this disclosure have specific activity at the hCRHR2 receptor and little activity at the hCRHR1 receptor.

[0368] [Table 3]

[0369] Similar to the polypeptide selectivity observed in hCRHR2 cells compared to hCRHR1 cells, the data presented in this table demonstrate that all polypeptides of this disclosure exhibit significantly higher efficacy in A7r5 cells (CRHR2 receptor) than in Y79 cells (CRHR1 receptor).

[0370] Example 5: Determination of association state by sedimentation velocity analysis using UltraCentrifugation (SV-AUC) SV-AUC was performed at 20°C using a Beckman Optima with an AN60 Ti rotor and a final rotor speed of 60,000 rpm. Peptides were dialyzed in 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 appropriate buffers as needed. Both samples and buffers were passed through a 0.22 μm PVDF centrifugation filter unit. The filtered samples were loaded into the sample sector of the cells. Only the filtered buffer was loaded into the reference sector of the cells. A wavelength scan was performed as a first step to select a suitable lambda (λ) value for a given compound having an OD of 1–1.2 at the tested concentration, which is generally 225 nm–254 nm, and was used in the Method Scan in the next step. The λ for each compound is shown in Table 1. Both absorbance and interference detection modules were used in the Method Scan. Temperature equilibrium was set as the first stage, 3 hours at a rotor speed of 0 rpm. Settlement velocity was set as the second stage, 60,000 rpm at a rotor speed. 300 scans were performed at a frequency of 60 seconds, or 500 scan counts at a frequency of 120 seconds, at the specified wavelength. The data were analyzed using SEDFIT software with a continuous c(s) distribution model applied. The settings for the relevant parameters are shown in Tables 4 and 5, and the settling coefficient (s) values ​​are reported in Svedherg units (S).

[0371] [Table 4]

[0372] Experiments to characterize the compound containing SEQ ID NO: 54 were performed at a frequency of 60 seconds and 300 scan counts, while experiments for all other polypeptides tested were performed at a frequency of 120 seconds and 500 scan counts.

[0373] Table 5. Settings for relevant parameters of the SEDFIT software using a continuous c(s) distribution model.

[0374] (a) Parameters used for the continuous sedimentation coefficient distribution c(s) in Sequence ID No. 54 [Table 5]

[0375] (b) Parameters used in other polypeptides in this disclosure. [Table 6]

[0376] Reference compound SEQ ID NO: 54 exhibited a sedimentation coefficient at all three concentrations tested (0.1 mg / mL, 1 mg / mL, and 10 mg / mL) indicating that higher-order oligomers were the dominant species. In contrast, the polypeptides of this disclosure were measured as having a lower self-association state compared to SEQ ID NO: 54.

[0377] Example 6. In vivo diet-induced obesity (DIO) model - chronic dose administration The DIO model represents a pre-diabetic state characterized by obesity. These animals, while not diabetic, exhibit insulin resistance, dyslipidemia, and hepatic steatohepatia after 12 weeks of a high-fat diet (60% Kcal from fat), all of which are characteristic of metabolic syndrome (Surwit RS et al., Diet-induced type II diabetes in C57BL / 6J mice. Diabetes 37(9):1163-7(1988)). Assessments of body weight change from baseline, fat mass change from baseline, lean body mass change from baseline, and fasting glucose or glucose change from vehicle in the same study are substantially described below. In the case of Sequence ID No. 55 combined with Sequence ID No. 25, tibialis anterior muscle weight at the end of the study is also reported as a measure of the effect on muscle weight.

[0378] Male C57BL6 mice aged 22–24 weeks (Jackson Laboratories 3800050; Bar Harbor, ME), fed a high-fat diet from 6 weeks of age, were housed one per cage and fed D12492 diet (60% lard high-fat diet: Research diets New Brunswick NJ) in a vivarium under normal photoperiod conditions prior to the start of the experiment. Block randomization was used to randomize animals to treatment groups based on body weight. On day 1 of the experiment, animals and food were weighed and recorded. The animals were divided into two equal groups and started on separate days (data combined) to simplify the study's logistics. Animals were administered a single subcutaneous injection (sc) of the indicated treatment in 40 mM Tris HCl pH 8.0 + 50 mM D-mannitol + 0.02% polysorbate 80 at a volume of 10 ml / kg on days 1 (start), 4, 7, 10, and 13 of the experiment. Vehicle control animals were injected with the same volume of this solution. The solution was kept in sterile capped vials stored at 4°C throughout the study period. Each treatment arm had 5-6 mice (n) per group.

[0379] body weight From day 1 to day 14 of the study, the animals' body weight was weighed daily before dose administration. This data was used to calculate the change in body weight. The animals were placed in a weighing pan, and the balance was stabilized. The weight was recorded.

[0380] Fasting glucose On day 14 of the experiment, the animals were fasted overnight (approximately 16-18 hours) by being placed in a clean cage with a clean wire rack, and given water but no food. On day 15, the animals were subjected to fasting blood glucose measurement as follows: blood was collected by removing the tail of the animal, and blood glucose was measured using an Accu-Chek Aviva glucose meter (Roche; Indianapolis, IN). Statistical significance ( *The p > 0 vs. 0 dose (one-way ANOVA Dunnett post-hoc test) was calculated using GraphPad Prism software (La Jolla, Ca).

[0381] body composition analysis Body composition was analyzed on day 1 and day 14 of the study using a quantitative nuclear magnetic resonance echoMRI analyzer (EMR-166-s, EchoMRI; Houston Tx). After calibrating the analyzer with a known amount of canola oil, the animals were placed in the analyzer and fat and lean (non-fat) mass were measured in grams. The change in mass was calculated by subtracting the value on day 14 from the value on day 1.

[0382] Tibialis anterior muscle weight On day 15 of the experiment, following tail hemorrhage due to blood glucose, the animals were euthanized by CO2 asphyxiation, and the right tibialis anterior muscle was incised. The muscle was cut from the fascia and tendon and weighed. The weight was recorded, and in the case of relative weight, the weight of the tibialis anterior in milligrams was divided by the body weight in grams.

[0383] Tables 6-11 below show the data corresponding to each of the above measurements. The data is expressed as arithmetic mean ± standard error of mean (SEM). Tables 6-9 show the ED50 for the listed parameters as appropriate. ED50 was calculated using the Find ECanything function with a top fixed to F=50 and vehicle mean in log(agonist) vs. response-GraphPad Prism software (La Jolla, Ca).

[0384] [Table 7] * = p<0.05 for the vehicle (one-way ANOVA, Dunnett's post-hoc test)

[0385] [Table 8] *= p<0.05 for the vehicle (one-way ANOVA, Dunnett's post-hoc test)

[0386] [Table 9] * = p<0.05 for the vehicle (one-way ANOVA, Dunnett's post-hoc test)

[0387] [Table 10] * = p<0.05 for the vehicle (one-way ANOVA, Dunnett's post-hoc test)

[0388] [Table 11] * = p<0.05 for vehicles from the same test (one-way ANOVA, Dunnett's post-hoc test)

[0389] [Table 12] * = p<0.05 for vehicle, #= p<0.05 for SEQ ID NO: 55. Tirzepatide 10 nmol / kg (one-way ANOVA, Dunnett's post-hoc test)

[0390] When the tested polypeptides were administered subcutaneously every 3 days for 14 consecutive days, the following significant differences were observed: (1) reduction in body weight, (2) reduction in fat mass, and (3) reduction in fasting blood glucose, in the absence of any significant change in lean body mass. Dose-response experiments for SEQ ID NO: 29, SEQ ID NO: 1, SEQ ID NO: 22, and SEQ ID NO: 25 demonstrated improved efficacy over the previous polypeptide SEQ ID NO: 54 in terms of (1) change in body weight and (2) change in fat mass, as measured by ED50 (Tables 6-9). Polypeptides including 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 demonstrated superior efficacy over the previous polypeptide SEQ ID NO: 54 in terms of (1) change in body weight and (2) change in fat mass at a single dose level of 30 nmol / kg (Table 10). As shown in Table 11, polypeptides containing SEQ ID NO: 25 enhanced both weight loss and fat loss induced by SEQ ID NO: 55 when combined with 10 nmol / kg at different dose levels, with statistically significant differences at lower doses of 30 nmol / kg of SEQ ID NO: 25 (for change in total weight) and 1 nmol / kg of SEQ ID NO: 25 (for change in fat mass). Compared to 10 nmol / kg of SEQ ID NO: 55 alone, polypeptides containing SEQ ID NO: 25 also increased both relative and absolute tibialis anterior muscle weight when combined with 10 nmol / kg at a low dose of 1 nmol / kg (Table 11).

[0391] Example 7: Pharmacokinetics biological analysis method The plasma concentrations of the compounds were measured by LC / MS. For each assay, the compound and the internal standard (IS) were extracted from 100% mouse, rat, pig, or monkey plasma (70 μL) using [50-50] isopropanol-methanol. When the compound and the IS contained in the supernatant were centrifuged, two distinct layers were formed. Aliquots (150 μL) of the supernatant were transferred to a Waters Sirocco Protein Precipitation Plate. The supernatant was eluted using a positive pressure manifold. The supernatant was mixed with 100-2hydrate-formic acid (400 μL) in a pre-treated Sep-Pak tC18 SPE plate, eluted, and discarded. The peptide and IS were eluted in two steps (50 μL each) with [100-2]acetonitrile-formic acid (100 μL). The eluate (40 μL) from the SPE plate was mixed with a dilution solution (1.875 μL of 5 × in vitrosol and 3.125 μL of 1% formic acid in water, 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 directed to a Thermo Fisher Scientific Orbitrap (Exploris 240) mass spectrometer for detection and quantification.

[0392] Pharmacokinetic studies Male and female cynomolgus monkeys were administered a single subcutaneous or intravenous dose (183-190 nmol / kg) of the compound in 40 mM Tris-HCl 5% D-mannitol (pH 8) at a volume of 0.5 mL / kg. For pharmacokinetic characterization, blood was collected from each animal at 0.5, 3, 6, 24, 48, 72, 96, 168, 240, 336, 408, and 504 hours after administration.

[0393] Male Yucatan miniature pigs were administered a single subcutaneous or intravenous dose (19 or 99 nmol / kg) of the compound in 40 mM Tris-HCl 5% D-mannitol (pH 8) at a volume of 0.01 or 0.05 mL / kg. For pharmacokinetic characterization, blood was 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.

[0394] Male Sprague Dawley rats were administered a single subcutaneous dose (183-201 nmol / kg) of the compound in 40 mM Tris-HCl 5% D-mannitol (pH 8) at a volume of 0.5 or 2 mL / kg. For pharmacokinetic characterization, blood was collected from each animal at 0.5, 3, 6, 24, 48, 72, 96, 120, and 168 hours post-administration.

[0395] Male Sprague Dawley rats were administered a single subcutaneous dose (10, 30, or 100 nmol / kg) of the compound in 40 mM Tris-HCl 5% D-mannitol (pH 8) at a volume of 1 mL / kg. For pharmacokinetic characterization, blood was collected from each animal at 3, 6, 24, 48, 72, 96, 120, and 168 hours post-administration.

[0396] Male CD-1 mice were administered a single subcutaneous dose (100 nmol / kg) of the compound in 40 mM Tris-HCl 5% D-mannitol (pH 8) at a volume of 5 mL / kg. For pharmacokinetic characterization, blood was collected from each animal at 6, 24, 48, 96, 168, 240, 288, and 336 hours post-administration.

[0397] [Table 13] Abbreviation:AUC 0-無限大 = Area under the curve from time 0 hours to infinity, CL / F = clearance / bioavailability, Tmax = time to maximum concentration, Cmax = maximum observed plasma concentration, T1 / 2 = half-life.

[0398] [Table 14] Abbreviation:AUC 0-無限大 = Area under the curve from time 0 to infinity, CL = clearance, C0 = estimated plasma concentration at time 0, T 1 / 2 = Half-life.

[0399] [Table 15] Abbreviation:AUC 0-無限大 = Area under the curve from time 0 hours to infinity, CL / F = clearance / bioavailability, Tmax = time to maximum concentration, Cmax = maximum observed plasma concentration, T1 / 2 = half-life.

[0400] [Table 16] Abbreviation:AUC 0-無限大 = Area under the curve from time 0 to infinity, CL = clearance, C0 = estimated plasma concentration at time 0, T 1 / 2 = Half-life.

[0401] [Table 17] Abbreviation:AUC 0-無限大 = Area under the curve from time 0 hours to infinity, CL / F = clearance / bioavailability, Tmax = time to maximum concentration, Cmax = maximum observed plasma concentration, T1 / 2 = half-life.

[0402] [Table 18] Abbreviation:AUC 0-無限大 = Area under the curve from time 0 hours to infinity, CL / F = clearance / bioavailability, Tmax = time to maximum concentration, Cmax = maximum observed plasma concentration, T1 / 2 = half-life.

[0403] [Table 19] Abbreviation:AUC 0-無限大 = Area under the curve from time 0 to infinity, CL / F = clearance / bioavailability, C0 = estimated plasma concentration at time 0, T 1 / 2 = Half-life.

[0404] [Table 20] Abbreviation:AUC 0-無限大 = Area under the curve from time 0 to infinity, CL / F = clearance / bioavailability, C0 = estimated plasma concentration at time 0, T1 / 2 = half-life.

[0405] [Table 21] Abbreviation:AUC 0-無限大 = Area under the curve from time 0 hours to infinity, CL / F = clearance / bioavailability, Tmax = time to maximum concentration, Cmax = maximum observed plasma concentration, T1 / 2 = half-life.

[0406] [Table 22] Abbreviation:AUC 0-無限大 = Area under the curve from time 0 to infinity, CL / F = clearance / biological availability, Tmax = time to maximum concentration, Cmax = maximum observed concentration, T1 / 2 = half-life.

[0407] PK conclusion Across different species, the PK profile supports a longer elimination half-life and reduced clearance compared to wild-type UCN2. These data indicate that the compounds described above have a pharmacokinetic profile suitable for once-weekly administration.

[0408] Example 8: Immunogenicity Risk Assessment MAPPS assay (MHC-related peptide proteomics) MAPPS characterizes MHC-II-presenting peptides on human dendritic cells treated with the test molecule. CD14+ cells isolated from normal human donor PBMCs were cultured and differentiated into immature DCs by incubation 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 to mature DCs. On day 6, the cells were lysed in RIPA buffer containing a protease inhibitor. Immunoprecipitation of the MHC-II complex was performed using a biotinylated anti-MHC-II antibody coupled to streptavidin beads. The bound complex was eluted and filtered. The isolated MHC-II peptide was analyzed by mass spectrometry. Peptide identification was generated by an internal proteomics pipeline using an enzyme-free search algorithm and a bovine / human database containing test sequences appended to the database. The KNIME workflow was used to process identification files from the samples. Peptides identified from the test samples were aligned relative to the parent sequence of the test molecule. The output was used to determine the percentage of donors presenting MHC-II peptides from the region of the test molecule. Polypeptides containing SEQ ID NOs. 54, 49, 50, 29, 41, 45, 1, 22, and 25 were tested in the MAPP assay. Polypeptides containing SEQ ID NOs. 41, 45, and 25 did not show peptides presented on the MHC-II complex in the assay. Compounds containing SEQ ID NOs. 54, 49, 50, 29, 1, and 22 presented at least one peptide cluster on the MHC-II complex in a subset of donors. Depending on the donor, up to three major clusters were observed, each slightly shifted from the others, spanning residues 1–33 in total.

[0409] T-cell proliferation assay This assay evaluated the ability of a test antibody or test MAPPS peptide to activate CD4+ T cells by inducing cell proliferation. PBMCs depleted of CD8+ T cells were prepared and labeled with CFSE. Each sample was tested with a medium control, keyhole limpet hemocyanin (KLH; positive clinical benchmark control), test antibody, or test MAPPS peptide. Cultures were incubated for 7 days. On day 7, samples were analyzed by flow cytometry. Reference analogs, including SEQ ID NO: 54, induced a response in 4 out of 10 donors, while analogs of this disclosure did not.

[0410] Existing reactivity (ACE assay format) This assay assessed the presence of pre-existing antibodies (ADAs) against a test molecule in untreated, normal human serum (NHS). Diluted NHS was incubated overnight on Pierce Streptavidin plates coated with biotinylated test molecules. The following day, captured binding proteins were acid-eluted and hard-coated onto Mesoscale (MSD) plates for detection using a combination of biotin-labeled molecules and ruthenium-labeled streptavidin. If anti-drug antibodies were present, they bound to the labeled drug, and the resulting signal was called a Tier 1 signal (represented as electrochemiluminescence). This signal was Tier 2, confirmed by adding an excess of unlabeled test molecules in the detection step (which suppressed the Tier 1 signal). The presence of pre-existing anti-drug antibodies was expressed as the magnitude of the 90th percentile of Tier 2 inhibition. The 90th percentile of Tier 2 inhibition is a statistical tool for assessing the magnitude of Tier 1 reactivity specificity. This 90th percentile was used to rank molecules in relation to ADA risk. The polypeptide containing SEQ ID NO: 1 exhibited higher 90th percentile tier 2 inhibition than the polypeptides containing SEQ ID NOs: 22 and 25.

[0411] [Table 23]

[0412] Abbreviations: ACE = Acid Capture and Elution; 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; MAPPs = MHC-related Peptide Proteomics; MHC = Major Histocompatibility Complex; MS = Mass Spectrometry; ND = No Data; T2 = Tier 2; TCEM = T Cell Exposure Motif; VH = Variable Heavy Chain; VL = Variable Light Chain; VHFR3 = Variable Heavy Chain Framework 3 Polypeptides containing SEQ ID NOs: 49, 50, 29, 41, 45, 1, 22, and 25 showed substantial improvement in immunogenicity compared to compounds containing SEQ ID NO: 54 in MAPP assays and / or T cell proliferation assays.

[0413] array Sequence ID 1 Ac-GSPSIVTS αMeL DVPT Aib αMeL LQKILEQERAEKARQQAK(γE-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-εK-CO-(CH2) 18 -CO2H)EAAEILAQV-NH2

[0414] Sequence ID 2 Ac-GSPSIVTS αMeL DVPT Aib αMeL LQKLLEQERAEKARQQAK(γE-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-εK-CO-(CH2) 18 -CO2H)EAAEILAQV-NH2

[0415] Sequence ID 3 Ac-GSPSIVTS αMeL DVPT Aib αMeL LQKILEQERAEKARQQAK((2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-PKE-γE-CO-(CH2) 18 -CO2H)EAAEILAQV-NH2

[0416] Sequence ID 4 Ac-GSPSIVTS αMeL DVPT Aib αMeL LQKILEQERAEKARQQAK(Ahx-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-γE-CO-(CH2) 18 -CO2H)EAAEILAQV-NH2

[0417] Sequence ID 5 Ac-GSPSIVTS αMeL DVPT Aib αMeL LQKILEQERAEKARQQAK(γE-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-Ahx-CO-(CH2) 18 -CO2H)EAAEILAQV-NH2

[0418] Sequence ID 6 Ac-GSPSIVTS αMeL DVPT Aib αMeL LQKILEQERAEKARQQAK(γE-GKEKEKE-CO-(CH2) 18 -CO2H)EAAEILAQV-NH2

[0419] Sequence ID 7 Ac-GSPSIVTS αMeL DVPT Aib αMeL LQKILEQERAEKARQQAK(G-(βA)3-γE-CO-(CH2) 18 -CO2H)EAAEILAQV-NH2

[0420] Sequence ID 8 Ac-GSPSIVTS αMeL DVPT Aib αMeL LQKILEQERAEKARQQAK(G-(βA)4-γE-CO-(CH2) 18 -CO2H)EAAEILAQV-NH2

[0421] Sequence ID 9 Ac-GSPSIVTS αMeL DVPT Aib αMeL LQKILEQERAEKARQQAK((2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-KEKEKE-γE-CO-(CH2)18 -CO2H)EAAEILAQV-NH2

[0422] Sequence ID 10 Ac-GSPSIVTS αMeL DVPT Aib αMeL LQKILEQERAEKARQQAK((βA)-S-(βA)-S-γE-CO-(CH2) 18 -CO2H)EAAEILAQV-NH2

[0423] Sequence ID 11 Ac-GSPSIVTS αMeL DVPT Aib αMeL LQKILEQERAEKARQQAK(γE-PKE-Ahx-CO-(CH2) 18 -CO2H)EAAEILAQV-NH2

[0424] Sequence ID 12 Ac-GSPSIVTS αMeL DVPT Aib αMeL LQKLLEQERAEKARQQAK(γE-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-εK-CO-(CH2) 16 -CO2H)EAAEILAQV-NH2

[0425] Sequence ID 13 Ac-GSPSIVTS αMeL DVPT Aib αMeL LQKILEQERAEKARQQAK(γE-PKE-εK-CO-(CH2) 18 -CO2H)EAAEILAQV-NH2

[0426] Sequence ID 14 Ac-GSPSIVTS αMeL DVPT Aib αMeL LQKILEQERAEKARQQAK(γE-PPP-εK-CO-(CH2) 18 -CO2H)EAAEILAQV-NH2

[0427] Sequence ID 15 Ac-GSPSIVTS αMeL DVPT Aib αMeL LQKILEQERAEKARQQAK(γE-PPPPPP-CO-(CH2) 18 -CO2H)EAAEILAQV-NH2

[0428] Sequence ID 16 Ac-GSPSIVTS αMeL DVPT Aib αMeL LQKILEQERAEKARQQAK(γE-PEPEPE-CO-(CH2) 18 -CO2H)EAAEILAQV-NH2

[0429] Sequence ID 17 Ac-GSPSIVTS αMeL DVPT Aib αMeL LQKILEQERAEKARQQAK(γE-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)2-εK-CO-(CH2) 18 -CO2H)EAAEILAQV-NH2

[0430] Sequence ID 18 Ac-GSPSIVTS αMeL DVPT Aib αMeL LQKILEQERAEKARQQAK(γE-εK-yE-εK-CO-(CH2) 18 -CO2H)EAAEILAQV-NH2

[0431] Sequence ID 19 Ac-GSPSIVTS αMeL DVPT Aib αMeL LQKILEQERAEKARQQAK(γE-εK-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-εK-CO-(CH2) 18 -CO2H)EAAEILAQV-NH2

[0432] Sequence ID 20 Ac-GSPSIVTS αMeL DVPT Aib αMeL LQKILEQERAEKARQQAKEAAEILAQV-NH2

[0433] Sequence ID 21 Ac-GSPSIVTS αMeL DVPT Aib αMeL LQKILEQERAEKARQQAK(γE- (2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-εK-CO-(CH2) 16 -CO2H)EAAEILAQV-NH2

[0434] Sequence ID 22 Pyr-GSPSGIVTS αMeL DVPT Aib αMeL LQKILEQERAEKARQQAK(γE-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-εK-CO-(CH2) 18 -CO2H)EAAEILAQV-NH2

[0435] Sequence ID 23 Pyr-GSPSGIVTS αMeL DVPT Aib αMeL LQKILEQERAEKARQQAKEAAEILAQV-NH2

[0436] Sequence ID 24 Pyr-GSPSGIVTS αMeL DVPT Aib αMeL LQKILEQERAEKARQQAK(γE-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-εK-CO-(CH2) 16 -CO2H)EAAEILAQV-NH2

[0437] Sequence ID 25 HO2C-(CH2) 18 -CO-εK-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-γE-GSPSIVTS αMeL DVPT Aib αMeL LQKILEQERAEKARQQATEAAEILAQV-NH2

[0438] Sequence ID 26 HO2C-(CH2) 18-CO-K-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-γE-GSPSIVTS αMeL DVPT Aib αMeL LQKILEQERAEKARQQATEAAEILAQV-NH2

[0439] Sequence ID 27 GSPSIVTS αMeL DVPT Aib αMeL LQKILEQERAEKARQQATEAAEILAQV-NH2

[0440] Sequence ID 28 HO2C-(CH2) 16 -CO-εK-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-γE-GSPSIVTS αMeL DVPT Aib αMeL LQKILEQERAEKARQQATEAAEILAQV-NH2

[0441] Sequence ID 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

[0442] Sequence ID 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

[0443] Sequence ID 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

[0444] Sequence ID 32 Ac-IVTS αMeL D (D-Val)PTG αMeL ALTERNATED((2-[2-(2-Diphenyl-Dipheric)-Dipheric]-Dipheral)-εK-γE-CO-(CH2) . 18 -CO2H)ENTHEILAQV-NH2

[0445] Sequence ID 33 Ac-IVTS αMeL D (D-Val)PTG αMeL LQUIRED((2-[2-(2-Diphenyl-Dipheric)-Dipheric]-Dipheral)-γE-εK-CO-(CH2) . 18 -CO2H)ENTHEILAQV-NH2

[0446] Sequence ID 34 Ac-IVTS αMeL D (D-Val)PTG αMeL LQUIRED(εK-(2-[2-(2-ν-ν)-ν]-ν)-γE-CO-(CH2); 18 -CO2H)ENTHEILAQV-NH2

[0447] Sequence ID 35 Ac-IVTS αMeL D (D-Val)PTG αMeL ALQUALIFIED(εK-γE-(2-[2-(2-ν-ν)-κν]-κν)-CO-(CH2); 18 -CO2H)ENTHEILAQV-NH2

[0448] Sequence ID 36 Ac-IVTS αMeL D (D-Val)PTG αMeL ALQUALIFIED(γE-(2-[2-(2-ν-ν)-ν]-ν)-κ-CO-(CH2); 18 -CO2H)ENTHEILAQV-NH2

[0449] Sequence ID 37 Ac-IVTS αMeL D (D-Val)PTG αMeL LQIKLEQERAEKARQQAK(γE-εK-(2-[2-(2-アミノ-エトキシ)-エトキシ]-アセチル)-CO-(CH2) 18 -CO2H)ENTEILAQV-NH2

[0450] Sequence ID 38 Ac-IVTS αMeL D (D-Val)PTG αMeL LQIKLEQERAEKARQQAK(γE-γE-εK-CO-(CH2) 18 -CO2H)ENTEILAQV-NH2

[0451] Sequence ID 39 Ac-IVTS αMeL D (D-Val)PTG αMeL LQIKLEQERAEKARQQAK(γE-εK-γE-CO-(CH2) 18 -CO2H)ENTEILAQV-NH2

[0452] Sequence ID 40 Ac-IVTS αMeL D (D-Val)PTG αMeL LQIKLEQERAEKARQQAK(εK-(γE)2-CO-(CH2) 18 -CO2H)ENTEILAQV-NH2

[0453] Sequence ID 41 Ac-IVTS αMeL D (D-Val)PT Aib αMeL LQIKLEQERAEKARQQAK((2-[2-(2-アミノ-エトキシ)-エトキシ]-アセチル)2-γE-CO-(CH2) 18 -CO2H)ENTEILAQV-NH2

[0454] Sequence ID 42 Ac-IVTS αMeL D (D-Val)PI Aib αMeL LQIKLEQERAEKARQQAK((2-[2-(2-アミノ-エトキシ)-エトキシ]-アセチル)2-γE-CO-(CH2) 18-CO2H)ENTHEILAQV-NH2

[0455] Sequence ID 43 Ac-IVTS αMeL D (D-Val)PTG αMeL(GKEKE-γE-CO-(CH2) 18 -CO2H)ENTHEILAQV-NH2

[0456] Sequence ID 44 Ac-IVTS αMeL D (D-Val)PTG αMeL ALTERNATED((2-[2-(2-Diphenyl-Dipheric)-Dipheric]-Dipheline)-(γE)2-CO-(CH2) . 18 -CO2H)ENTHEILAQV-NH2

[0457] Sequence ID 45 THE Aib αMeL IN THE αMeL D (D-Val) OF THE γE-GPSIVTS WAS CONTAINED((2-[2-(2-ノ-ツシ)-。・]-ア・)2-γE-CO-(CH2) . 18 -CO2H)ENTHEILAQV-NH2

[0458] Sequence ID 46 Ac-PPPIVTS αMeL D (D-Val)PT Aib αMeL LQK Aib LEATHER(γE-(2-[2-(2-ツ-κν)-κν]-ν)-κ-CO-(CH2); 18 -CO2H)ENTHEILAQV-NH2

[0459] Sequence ID 47 Ac-PPPIVTS αMeL D (D-Val)PT Aib αMeL LQGROUP(γE-(2-[2-(2-ν-ν)-κν]-ν)-εK-CO-(CH2); 18 -CO2H)EAAEILAQV-NH2

[0460] Sequence ID 48 Ac-PPPIVTS αMeL DVPT Aib αMeL LQKLLEQERAEKARQQAK(γE-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-εK-CO-(CH2) 18 -CO2H)EAAEILAQV-NH2

[0461] Sequence ID 49 (NMeI)VTSLDVPIG αMeL LQKLIEQEKQEKEKQQAK((2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)2-γE-CO-(CH2) 18 -CO2H)ENTEILAQV-NH2

[0462] Sequence ID 50 (NMeI)VTSLDVPIG αMeL LQKLLEQERAEKARQQAK((2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)2-γE-CO-(CH2) 18 -CO2H)EAAEILAQV-NH2

[0463] Sequence ID 51 Ac-IVTS αMeL D (D-Val)PI Aib αMeL LQIKLEQERAEKARQQAK((γE)2-CO-(CH2) 18 -CO2H)ENTEILAQV-NH2

[0464] Sequence ID 52 HO2C-(CH2) 18 -CO-Dap-(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-γE-GSPSIVTS αMeL DVPT Aib αMeL LEKILEQERAEKARQQATEAAEILAQV-NH2

[0465] Sequence ID 53 IVLSLDVPIGLLQILLEQARARAAREQATTNARILARV-NH2

[0466] Sequence ID 54 (NMeI)VLSLDVPIGLLQILLEQEKQEKEKQQAK((2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)2-(γE)2-CO-(CH2) 18 -CO2H)TNAQILAQV-NH2

[0467] Sequence ID 55 Y Aib EGTFTSDYSI Aib LDKIAQK((2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)2-γE-CO-(CH2) 18 -CO2H)AFVQWLIAGGPSSGAPPPS-NH2

[0468] Sequence ID 56 Y Aib QGTFTSDYSI αMeL LDKK((2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)2-γE-CO-(CH2) 18 -CO2H)AQ Aib AFIEYLLEGGPSSGAPPPS-NH2

[0469] Sequence ID 57 Y Aib QGTFTSDYSI αMeL LDKK((2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-γE-CO-(CH2) 18 -CO2H)AQ Aib AFIEYLLEGGPSSGAPPPS-NH2

[0470] Sequence ID 58 Y Aib QGTFTSDYSILLDKK((2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)2-γE-CO-(CH2) 18 -CO2H)AQ Aib AFIEYLLEGGPSSGAPPPS-NH2

[0471] Sequence ID 59 H Aib QGTFTSDYSKYLDEKKAK((2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)2-γE-CO-(CH2) 16 -CO2H)EFVEWLLEGGPSSG-NH2

[0472] Sequence ID 60 H Aib QGTFTSDYSKYLDEKKAK((2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)2-(γE)2-CO-(CH2) 16 -CO2H)EFVEWLLSGGPSSG-NH2

[0473] Sequence ID 61 H Aib QGTFTSDYSKYLDEKKAK((2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)2-γE-CO-(CH2) 18 -CO2H)EFVEWLLEGGPSSG-NH2

[0474] Sequence ID 62 H Aib QGTFTSDYSKYLDEKKAK((2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)2-(γE)2-CO-(CH2) 18 -CO2H)EFVEWLLSGGPSSG-NH2

[0475] Sequence ID 63 γE-[CNTATC] * ATG Orn LAE αMeF LVRSSN (NMeN)FGPK((γE)2-CO-(CH2) 18 -CO2H)LPPTEVGSNTY-NH2

[0476] Sequence ID 64 γE-[CNTATC] * ATG Orn LAE αMeF LVRSSN (NMeD)FGPK((γE)2-CO-(CH2) 18 -CO2H)LPPTEVGSNTY-NH2

[0477] Sequence ID 65 K((γE)2-CO-(CH2) 18 -CO2H)-[CETATC] * ATG Orn LAE αMeF LVRSSN (NMeD)FGPILPPTEVGSNTY-NH2

[0478] Sequence ID 66 K((γE)2-CO-(CH2) 18 -CO2H)-[CETATC] * ATG Orn αMeL AEFLVRSSHNFGPILPPTEVGSNTY-NH2

[0479] Sequence ID 67 Ac-ASHLSTAVLGK((2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)2-γE-CO-(CH2) 18 -CO2H)LS Aib ELHKLEDYPRTDVGAESP-NH2

[0480] Sequence ID 68 X1X2X3X4X5X6IVTSX 11 DX 13 PX 15 X 16 X 17 LX 19 X 20 X 21 X 22 EQEX 26 X 27 EKX 30 X 31 QQAX 35 EX 37 X 38 EILAQV, X1 is either Pyr, E, γE, or absent. X2 is either G or does not exist. X3 is either G, S, or does not exist. X4 is either S, P, G, or does not exist. X5 is either S, P, or does not exist. X6 is either G, S, or P, or does not exist. X 11 However, it is L or αMeL, X 13 However, it is V or D-Val, X 15 However, it is T or I, X 16 However, it is Aib or G, X 17 However, it is L or αMeL, X 19 However, it is either Q or E, X 20 However, it is K or I, X 21 However, it is I, K, Aib, or L. X 22 However, it is either I or L. X 26 However, it is R or K, X 27 However, it is either A or Q, X 30 However, it is A or E, X 31 However, it is R or K, X 35 However, it is either K or T, X 37 However, it is either A or N, X 38 However, it is A or T, Polypeptides or pharmaceutically acceptable salts thereof are provided, wherein the N-terminal amino acid is optionally N-acylated, N-acetylated, or N-methylated, and the C-terminal amino acid is optionally amidated.

[0481] Sequence ID 69 GSPSIVTS αMeL DVPT Aib αMeL LQKX 21 LEQERAEKARQQAKEAAEILAQV During the ceremony, X 21 However, it is either I or L. The N-terminal amino acid is optionally N-acetylated, and the C-terminal amino acid is optionally amidated.

[0482] Sequence ID 70 Pyr GSPSGIVTS αMeL DVPT Aib αMeL LQKILEQERAEKARQQAKEAAEILAQV The C-terminal amino acid is optionally amidated.

[0483] Sequence ID 71 X1X2GSPSIVTS αMeL DVPT Aib αMeLLX 19 KILEQERAEKARQQATEAAEILAQV, X1 is either γE or does not exist. X2 does not exist. X 19 However, it is either Q or E, The C-terminal amino acid is optionally amidated.

[0484] Sequence ID 72 IVTS αMeL D D-Val PX 15 X 16 αMeLLQIKLEQERAEKARQQAKENTEILAQV (Sequence ID 72), X 15 However, it is T or I, X 16 However, it is G or Aib, The N-terminus is an amino acid that is optionally N-acetylated, and the C-terminal amino acid is optionally amidated.

[0485] Sequence ID 73 PPPIVTS αMeL DX 13 PTAib αMeL LQKX 21 LEQERAEKARQQAKEX 37 X 38 EILAQV, X 13 However, it is V or D-Val, X 21 However, it is L or Aib, X37 However, it is either A or N, X 38 However, it is A or T, The N-terminal amino acid is optionally N-acetylated, and the C-terminal amino acid is optionally amidated.

[0486] Sequence ID 74 γE-GKEKEKE-CO-(CH2) 18 -CO2H

[0487] Sequence ID 75 (2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-KEKEKE-γE-CO-(CH2) 18 -CO2H

[0488] Sequence ID 76 γE-PPPPPP-CO-(CH2) 18 -CO2H

[0489] Sequence ID 77 γE-PEPEPE-CO-(CH2) 18 -CO2H

[0490] Sequence ID 78 GKEKEKE-γE-CO-(CH2) 18 -CO2H

[0491] Sequence ID 79 H Aib EGTFTSDVSSYLEGQAAK((2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)2-γE-CO-(CH2) 16 -CO2H)EFIAWLVRGRG-OH

[0492] Sequence ID 80 HAEGTFTSDVSSYLEGQAAK(γE-CO-(CH2) 14 -CH3)EFIAWLVRGRG-OH

[0493] Sequence ID 81 Dulaglutide is a human GLP-1 receptor agonist containing a GLP-1 analog dimer, fused at its C-terminus to the N-terminus of an immunoglobulin Fc portion analog via a peptide linker, and is identified as CAS Registry No. 923950-08-7, providing the following chemical name: 7-37-glucagon-like peptide I[8-glycine, 22-glutamic acid, 36-glycine](synthetic human) fusion protein, dimer, containing immunoglobulin G4(synthetic human Fc fragment), peptide(synthetic 16-amino acid linker). Each monomer of dulaglutide has the amino acid sequence described in SEQ ID NO: 81: HGEGTFTSDVSSYLEEQAAKEFIAWLVKGGGGGGGSGGGGGSGGGGSAESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTV LHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG

Claims

1. A polypeptide having activity at the corticotropin-releasing hormone receptor-2 (CRH2), or a pharmaceutically acceptable salt thereof, X １ X ２ X ３ X ４ X ５ X ６ IVTSX １１ DX １３ PX １５ X １６ X １７ LX １９ X ２０ X ２１ X ２２ EQEX ２６ X ２７ EKX ３０ X ３１ QQAX ３５ EX ３７ X ３８ comprises EILAQV (SEQ ID NO: 68), a) X １ However, it does not exist, X ２ However, it does not exist, X ３ However, G is X ４ However, S is X ５ However, P is X ６ However, S is X １１ However, it is αMeL, and X １３ However, V is X １５ However, T is X １６ However, it is Aib, and X １７ However, it is αMeL, and X １９ However, Q is X ２０ However, K is X ２１ However, it is either I or L, and X ２２ However, L is X ２６ However, R is X ２７ However, A is X ３０ However, A is X ３１ However, R is X ３５ However, K is X ３７ However, A is X ３８ However, is it A, or b) X １ However, it is either γE or does not exist, X ２ However, it does not exist, X ３ However, G is X ４ However, S is X ５ However, P is X ６ However, S is X １１ However, it is αMeL, and X １３ However, V is X １５ However, T is X １６ However, it is Aib, and X １７ However, it is αMeL, and X １９ However, it is either Q or E, and X ２０ However, K is X ２１ However, I is X ２２ However, L is X ２６ However, R is X ２７ However, A is X ３０ However, A is X ３１ However, R is X ３５ However, T is X ３７ However, A is X ３８ However, A is Polypeptides, or pharmaceutically acceptable salts thereof.

2. X １ However, it does not exist, X ２ However, it does not exist, X ３ However, G is X ４ However, S is X ５ However, P is X ６ However, S is X １１ However, it is αMeL, and X １３ However, V is X １５ However, T is X １６ However, it is Aib, and X １７ However, it is αMeL, and X １９ However, Q is X ２０ However, K is X ２１ However, it is I or L, and X ２２ However, L is X ２６ However, R is X ２７ However, A is X ３０ However, A is X ３１ However, R is X ３５ However, K is X ３７ However, A is X ３８ However, A is the polypeptide according to claim 1, or a pharmaceutically acceptable salt thereof.

3. X ２１ The polypeptide according to claim 2, or a pharmaceutically acceptable salt thereof, wherein I.

4. X ２１ The polypeptide according to claim 2, or a pharmaceutically acceptable salt thereof, wherein X is L.

5. X １ is γE or is absent, and X ２ is absent, and X ３ is G, and X ４ is S, and X ５ is P, and X ６ is S, and X １１ is αMeL, and X １３ is V, and X １５ is T, and X １６ is Aib, and X １７ is αMeL, and X １９ is Q or E, and X ２０ is K, and X ２１ is I, and X ２２ is L, and X ２６ is R, and X ２７ is A, and X ３０ is A, and X ３１ is R, and X ３５ is T, and X ３７ is A, and X ３８ is A, the polypeptide according to claim 1, or a pharmaceutically acceptable salt thereof.

6. X １９ Q is the polypeptide according to claim 5, or a pharmaceutically acceptable salt thereof.

7. X １９ The polypeptide according to claim 5, or a pharmaceutically acceptable salt thereof, wherein E is present.

8. A polypeptide according to any one of claims 1 to 7, further comprising a fatty acid, wherein the fatty acid conjugates with the polypeptide at an amino acid having a functional group available for conjugation, either via direct bonding or via a linker between the amino acid and the fatty acid, or a pharmaceutically acceptable salt thereof.

9. A polypeptide according to any one of claims 1 to 7, or a pharmaceutically acceptable salt thereof, wherein the N-terminal amino acid is N-acylated via a fatty acid conjugated with the N-terminal amino acid, via direct bonding, or via a linker between the N-terminal amino acid and the fatty acid.

10. A polypeptide according to any one of claims 1 to 7, wherein the N-terminal amino acid is N-acetylated, or a pharmaceutically acceptable salt thereof.

11. A polypeptide according to any one of claims 1 to 7, wherein the N-terminal amino acid is N-methylated, or a pharmaceutically acceptable salt thereof.

12. A polypeptide according to any one of claims 1 to 7, wherein the C-terminal amino acid is amidated, or a pharmaceutically acceptable salt thereof.

13. A polypeptide, or a pharmaceutically acceptable salt thereof, comprising an amino acid sequence selected from any one of SEQ ID NOs: 1-21, 25, 28, or 52.

14. A pharmaceutical composition comprising a polypeptide according to any one of claims 1 to 7 or 13, or a pharmaceutically acceptable salt thereof, and at least one pharmaceutically acceptable carrier, diluent, or excipient.

15. A drug for treating a disease or condition selected from the group consisting of diabetes mellitus, 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), comprising a polypeptide according to any one of claims 1 to 7 or 13, or a pharmaceutically acceptable salt thereof.

16. Use of a polypeptide according to any one of claims 1 to 7 or 13, or a pharmaceutically acceptable salt thereof, in the manufacture of a pharmaceutical product for treating a disease or condition selected from the group consisting of diabetes mellitus, 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).

17. A drug comprising a polypeptide according to any one of claims 1 to 7 or 13, or a pharmaceutically acceptable salt thereof, for treating a disease or condition selected from the group consisting of diabetes mellitus, 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), characterized in that it is used in combination with an additional therapeutic agent.

18. The agent according to claim 17, wherein the additional therapeutic agent is a dual agonist of GIP and GLP-1 receptors.

19. The agent according to claim 18, wherein the dual agonist of the GIP and GLP-1 receptor is a compound comprising SEQ ID NO:

55.

20. The agent according to claim 17, wherein the additional therapeutic agent is a GLP-1 receptor agonist.

21. The agent according to claim 20, wherein the GLP-1 receptor agonist is a compound comprising the amino acid sequence of SEQ ID NO: 79, SEQ ID NO: 80, or SEQ ID NO: 81.

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