Long-acting GLP-1 / GIP dual agonist

Abstract

The present invention relates to long-acting glucagon-like peptide-1 and human glucose-dependent insulinotropic polypeptide (GIP) agonist polypeptides that may be useful in the treatment of type 2 diabetes (T2D), diabetes associated with obesity, obesity, and hyperlipidemia.

Description

[Technical Field]

[0001] The present invention relates to a long-acting glucagon-like peptide-1 and a human glucose-dependent insulinotropic polypeptide / gastrointestinal peptide (GIP) agonist polypeptide that may be useful in the treatment of type 2 diabetes (T2D), diabetes with obesity, obesity, and hyperlipidemia. [Background technology]

[0002] Treatment of type 2 diabetes (T2DM) with glucagon-like peptide-1 receptor agonists (GLP-1RAs) results in improved glycemic control, weight loss, and improvement of several cardiovascular risk factors. These benefits are mediated by the glucagon-like peptide-1 receptor (GLP-1R), a member of the class B family of G protein-coupled receptors, expressed in pancreatic beta cells, various cell types of the gastrointestinal tract, and neurons in both the central (CNS) and peripheral nervous systems. Activation of GLP-1R signaling by GLP-1RAs improves glucose homeostasis by enhancing glucose-stimulated insulin secretion, slowing gastric emptying, and decreasing plasma glucagon levels, and reduces weight by activating the anorexia pathway in the brain. Due to the glucose dependence of beta cell activation, GLP-1RAs are not associated with an increased risk of hypoglycemia. The broad metabolic benefits of GLP-1RAs have established this class in the T2DM treatment paradigm; however, many patients fail to reach their HbA1c / glycemic targets, and the weight loss achieved with these drugs, therefore, requires higher doses, which also increases the risk of adverse events in the glycemic index (GI), remaining far lower than what can be achieved with bariatric surgery, the most powerful clinical intervention for obesity. Thus, there is a great opportunity to improve existing treatments for the GLP-1RA class.

[0003] One new approach is to combine a basic GLP-1RA therapy with pharmacological strategies targeting additional pathways involved in nutrition and energy metabolism, such as glucose-dependent insulinotropic polypeptide (GIP). GIP is an incretin secreted from K cells in the upper small intestine, duodenum, in response to food. Under normal physiological conditions, postprandial GIP levels are approximately four-fold higher compared to GLP-1. GIP bears the majority of the insulinotropic incretin effect in humans and has important additional functions distinct from GLP-1. Unlike GLP-1, GIP is both glucagonotropic and insulinotropic in a glucose-dependent manner, stimulating glucagon secretion in a dose-dependent manner under hypoglycemic conditions and insulin secretion under hyperglycemic conditions, and the released glucagon promotes insulin secretion. Both the GIP receptor (GIPR) and the GLP-1 receptor are present on beta cells, but GIPR is abundant in adipose tissue and is found in many non-overlapping regions of the CNS, so GIPR expression is differentially distributed in extrapancreatic tissues. GIP is involved in adipose tissue carbohydrate and lipid metabolism by virtue of its actions controlling glucose uptake, lipolysis, and lipoprotein lipase activity. These findings suggest that pharmacological activation of GIPR may have therapeutic benefits in peripheral energy metabolism. Recently, single-molecule multifunctional peptides combining GLP-1RA activity and GIP activity have been proposed as new therapeutic agents for blood glucose and body weight management.

[0004] U.S. Patent No. 9,474,780 discloses dual GLP-1 and GIP receptor agonists comprising tirzepatide.

Chemical formula

[0005] Tirzepatide is undergoing Phase III clinical trials for T2DM and obesity.

[0006] WIPO publication numbers WO201774714A1, WO202023386A1, WO2020023388A1, WO2015067715A2, WO2016111971A1, and WO2013164483A1 disclose GLP-1R and GIPR dual agonist compounds. [Overview of the Initiative]

[0007] The present invention provides a polypeptide comprising the following amino acid sequence or a pharmaceutically acceptable salt thereof: Y-X1-EGTFTSDYSI-X2-L-Xaa15-KIA-Xaa19-X3-Xaa21-FV-Xaa24-WL-X4-AGGPSSGAPPPS-X5-X6-X7-X8-X9-X10-X11 (Sequence number 1) In the formula, X1 is Aib, Ser(OMe), or (D)Ser(OMe), X2 is Tyr, Ser(OMe), (D)Ser(OMe), or Aib. X3 is either Gln or Lys, and in the formula, if X3 is Lys, the amino(ε-amino) group in the side chain of Lys is acylated at the following part: {―UWYZ In the formula, U is -C(O)-CH2-O-(CH2)2-O-(CH2)2-NH-}, where} is the attachment point to the group W. W is -C(O)-NH-(CH2) p Selected from the group consisting of -NH-], -C(O)-C(CH3)2-NH-], and -C(O)-CH2-O-(CH2)2-O-(CH2)2-NH-], where p is 3 or 4, and where ] is the attachment point to group Y. Y is -C(O)-(CH2)2-CH(COOH)NH--, and -- is the attachment point to group Z. Z is -C(O)-(CH2). n -COOH or -C(O)-(CH2) n -CH3, where n is an integer between 14 and 20. However, if XX3 is Lys and X2 is Aib, then W is not -C(O)-CH2-O-(CH2)2-O-(CH2)2-NH-], X4 is Leu, Ile, or Glu. X5 is either absent or is Arg or Lys. If X5 is Lys, the amino(ε-amino) side chain of Lys is acylated at the following point: {―U'- W'-Y'-Z' In the formula, U' is -C(O)-CH2-O-(CH2)2-O-(CH2)2-NH-}, where} is the attachment point to the group W'. W' is -C(O)-NH-(CH2) q Selected from the group consisting of -NH-], -C(O)-C(CH3)2-NH-], and -C(O)-CH2-O-(CH2)2-O-(CH2)2-NH-], where q is 3 or 4, and in the formula, ] is the attachment point with group Y'. Y' is -C(O)-(CH2)2-CH(COOH)NH--, and -- is the attachment point to group Z'. Z' is -C(O)-(CH2). m -COOH or -C(O)-(CH2) m -CH3, where m is an integer between 14 and 20. X6 does not exist, or it is Lys. X7 does not exist, or it is Lys. X8 does not exist, or it is Lys. X9 does not exist, or it is Lys. X10 does not exist, or it is Lys. X11 does not exist, or it is Lys. Xaa15 is either Asp or Glu. Xaa19 is either Gln or Ala. Xaa21 is either Ala or Glu. Xaa24 is either Gln or Asn. In the formula, the acid group of the C-terminal amino acid is either a free carboxylic acid group or is amidated as a C-terminal primary amide. However, at least one of X3 and X5 is Lys.

[0008] Abbreviation A1: 2-aminoisobutyric acid DIPEA: N,N'-di-isopropylethylamine HOBt: 1-hydroxybenzotriazole DIPC: N,N'-di-isopropylcarbodiimide THF: Tetrahydrofuran DCM: Dichloromethane DMAP: 4-dimethylaminopyridine DIC: Diisopropylcarbodiimide DMAc: Dimethylacetamide [Modes for carrying out the invention]

[0009] This invention provides a stable, long-acting GLP-1 / GIP agonist polypeptide that may be useful in the treatment of type 2 diabetes (T2D), diabetes with obesity, obesity, and hyperlipidemia. The polypeptide of this invention is considered to be long-acting, which may eliminate the need for frequent administration to patients who require it.

[0010] Accordingly, in one embodiment, the present invention provides a polypeptide comprising the following amino acid sequence or a pharmaceutically acceptable salt thereof: Y-X1-EGTFTSDYSI-X2-L-Xaa15-KIA-Xaa19-X3-Xaa21-FV-Xaa24-WL-X4-AGGPSSGAPPPS-X5-X6-X7-X8-X9-X10-X11 (Sequence number 1) In the formula, X1 is Aib, Ser(OMe), or (D)Ser(OMe), X2 is Tyr, Ser(OMe), (D)Ser(OMe), or Aib. X3 is either Gln or Lys, and in the formula, if X3 is Lys, the amino(ε-amino) group in the side chain of Lys is acylated at the following part: {―UWYZ In the formula, U is -C(O)-CH2-O-(CH2)2-O-(CH2)2-NH-}, where} is the attachment point to group W, W is -C(O)-NH-(CH2) p -NH-], -C(O)-C(CH3)2-NH-], and -C(O)-CH2-O-(CH2)2-O-(CH2)2-NH-], and is selected from the group consisting of, where p is 3 or 4, and where ] is the attachment point to group Y, Y is -C(O)-(CH2)2-CH(COOH)NH--, and -- is the attachment point to group Z, Z is -C(O)-(CH2) n -COOH or -C(O)-(CH2) n -CH3, where n is an integer from 14 to 20, However, when XX3 is Lys and X2 is Aib, W is not -C(O)-CH2-O-(CH2)2-O-(CH2)2-NH-], X4 is Leu, Ile or Glu, X5 does not exist or is Arg or Lys. When X5 is Lys, the side-chain amino (ε-amino) group of Lys is acylated in the following moiety, {―U’-W’-Y’-Z’ In the formula, U’ is -C(O)-CH2-O-(CH2)2-O-(CH2)2-NH-}, where} is the attachment point to group W’, W’ is -C(O)-NH-(CH2) q -NH-], -C(O)-C(CH3)2-NH-], and -C(O)-CH2-O-(CH2)2-O-(CH2)2-NH-], and is selected from the group consisting of, where p is 3 or 4, and where 〕 is the attachment point to group Y’, Y’ is -C(O)-(CH2)2-CH(COOH)NH--, and -- is the attachment point to group Z’, Z’ is -C(O)-(CH2) m -COOH or -C(O)-(CH2) m -CH3, where m is an integer from 14 to 20, X6 does not exist or is Lys, X7 does not exist, or it is Lys. X8 does not exist, or it is Lys. X9 does not exist, or it is Lys. X10 does not exist, or it is Lys. X11 does not exist, or it is Lys. Xaa15 is either Asp or Glu. Xaa19 is either Gln or Ala. Xaa21 is either Ala or Glu. Xaa24 is either Gln or Asn. In the formula, the acid group of the C-terminal amino acid is either a free carboxylic acid group or is amidated as a C-terminal primary amide. However, at least one of X3 and X5 is Lys.

[0011] In one embodiment of the present invention, X1 is Aib.

[0012] In another embodiment of the present invention, X2 is Aib.

[0013] In another embodiment of the present invention, both X1 and X2 are Aib.

[0014] In another embodiment of the present invention, X1 is Aib and X2 is Ser(OMe) or (D)Ser(OMe).

[0015] In another embodiment of the present invention, X1 is Ser(OMe) or (D)Ser(OMe), and X2 is Aib.

[0016] In another embodiment of the present invention, X4 is Leu or Ile.

[0017] In another embodiment of the present invention, X4 is Ile.

[0018] In another embodiment of the present invention, X5 is Lys or Arg.

[0019] In another embodiment of the present invention, X3 is Lys, and X5 is absent or Arg.

[0020] In another embodiment of the present invention, X3 is Gln and X5 is Lys.

[0021] In another embodiment of the present invention, W is -C(O)-C(CH3)2-NH-].

[0022] In another embodiment of the present invention, W is -C(O)-NH-(CH2) p -NH-], where p is 3 or 4.

[0023] In another embodiment of the present invention, W is -C(O)-NH-(CH2)4-NH-].

[0024] In another embodiment of the present invention, W is -C(O)-CH2-O-(CH2)2-O-(CH2)2-NH-].

[0025] In another embodiment of the present invention, W' is -C(O)-C(CH3)2-NH-].

[0026] In another embodiment of the present invention, W' is -C(O)-NH-(CH2) q -NH-], where p is 3 or 4.

[0027] In another embodiment of the present invention, W' is -C(O)-NH-(CH2)4-NH-].

[0028] In another embodiment of the present invention, W' is -C(O)-CH2-O-(CH2)2-O-(CH2)2-NH-].

[0029] In another embodiment of the present invention, the C-terminal amino acid is amidated as a C-terminal primary amide.

[0030] In another embodiment of the present invention, the acid group of the C-terminal amino acid is a free carboxylic acid.

[0031] In another embodiment of the present invention, n is 16, 17, 18, 19, or 20. In a preferred embodiment, n is 18 or 20. In yet another preferred embodiment, n is 20. In another preferred embodiment, n is 16 or 18. In yet another preferred embodiment, n is 18.

[0032] In another embodiment of the present invention, Z is -C(O)-(CH2) n It is -COOH, and n is 16 or 18.

[0033] In another embodiment of the present invention, m is 16, 17, 18, 19, or 20. In a preferred embodiment, m is 18 or 20. In yet another preferred embodiment, m is 20. In another preferred embodiment, m is 16 or 18. In yet another preferred embodiment, m is 18.

[0034] In another embodiment of the present invention, Z' is -C(O)-(CH2) m It is -COOH, and m is 16 or 18.

[0035] In another embodiment of the present invention, W is -C(O)-NH-(CH2)4-NH- and Z is -C(O)-(CH2) n It is -COOH, and n is 18.

[0036] In another embodiment of the present invention, W is -C(O)-C(CH3)2-NH- and Z is -C(O)-(CH2) n It is -COOH, and n is 16.

[0037] In another embodiment of the present invention, W is -C(O)-C(CH3)2-NH- and Z is -C(O)-(CH2) n It is -COOH, and n is 18.

[0038] In another embodiment of the present invention, W is -C(O)-CH2-O-(CH2)2-O-(CH2)2-NH-] and Z is -C(O)-(CH2) n It is -COOH, and n is 16.

[0039] In another embodiment of the present invention, W is -C(O)-CH2-O-(CH2)2-O-(CH2)2-NH-] and Z is -C(O)-(CH2) n It is -COOH, and n is 18.

[0040] In another embodiment of the present invention, W' is -C(O)-NH-(CH2)4-NH- and Z' is -C(O)-(CH2) m It is -COOH, and m is 18.

[0041] In another embodiment of the present invention, W' is -C(O)-C(CH3)2-NH- and Z' is -C(O)-(CH2) m It is -COOH, and m is 16.

[0042] In another embodiment of the present invention, W' is -C(O)-C(CH3)2-NH- and Z' is -C(O)-(CH2) m It is -COOH, and m is 18.

[0043] In another embodiment of the present invention, W' is -C(O)-CH2-O-(CH2)2-O-(CH2)2-NH-] and Z' is -C(O)-(CH2) m It is -COOH, and m is 16.

[0044] In another embodiment of the present invention, W' is -C(O)-CH2-O-(CH2)2-O-(CH2)2-NH-] and Z' is -C(O)-(CH2) m It is -COOH, and m is 18.

[0045] In another embodiment of the present invention, X5, X6, X7, X8, X9, X10, and X11 are all absent.

[0046] In another embodiment of the present invention, Xaa15 is Asp.

[0047] In another embodiment of the present invention, Xaa19 is Gln.

[0048] In another embodiment of the present invention, Xaa21 is Ala.

[0049] In another embodiment of the present invention, Xaa24 is Gln.

[0050] In another embodiment of the present invention, X1 is Aib, and X2 is Ser(OMe) or Tyr.

[0051] In another embodiment of the present invention, X1 is Aib and X2 is Ser(OMe).

[0052] In another embodiment of the present invention, X1 is Aib and X2 is Tyr.

[0053] In another embodiment of the present invention, X3 is Gln.

[0054] In another embodiment of the present invention, X4 is Leu.

[0055] In another embodiment of the present invention, X5 is Lys, where the side-chain amino(ε-amino) group of Lys is acylated at the following part. {―U'- W'-Y'-Z'

[0056] In another embodiment of the present invention, W' is -C(O)-C(CH3)2-NH- and Z' is -C(O)(CH2) m It is -COOH, and m is 18.

[0057] In another embodiment of the present invention, Xaa15 is Glu.

[0058] In another embodiment of the present invention, Xaa19 is Ala.

[0059] In another embodiment of the present invention, Xaa21 is Glu.

[0060] In another embodiment of the present invention, Xaa24 is Asn.

[0061] In another embodiment of the present invention, X6, X7, X8, X9, X10, and X11 are all absent.

[0062] In another embodiment, the present invention provides a polypeptide comprising the following amino acid sequence or a pharmaceutically acceptable salt thereof, Y-Aib-EGTFTSDYSI-Ser(OMe)-LDKIAQ-X3-AFVQWL-X4-AGGPSSGAPPPS-X5-X6-X7-X8-X9-X10-X11 (SEQ ID NO: 2), During the ceremony X3 is Lys, and in the formula, the amino(ε-amino) group of the Lys side chain is acylated at the following part: {―UWYZ In the formula, U is -C(O)-CH2-O-(CH2)2-O-(CH2)2-NH-}, where} is the attachment point to the group W. W is -C(O)-NH-(CH2) p Selected from the group consisting of -NH-], -C(O)-C(CH3)2-NH-], and -C(O)-CH2-O-(CH2)2-O-(CH2)2-NH-], where p is 3 or 4, and where ] is the attachment point to group Y. Y is -C(O)-(CH2)2-CH(COOH)NH--, and -- is the attachment point to group Z. Z is -C(O)-(CH2). n -COOH or -C(O)-(CH2) n -CH3, where n is an integer between 14 and 20. X4 is either Ile or Glu. X5 does not exist, or it is Arg. X6 does not exist, or it is Lys. X7 does not exist, or it is Lys. X8 does not exist, or it is Lys. X9 does not exist, or it is Lys. X10 does not exist, or it is Lys. X11 does not exist, or it is Lys. In the formula, the acid group of the C-terminal amino acid is either a free carboxylic acid group or is amidated as a C-terminal primary amide.

[0063] In one embodiment of the present invention, X4 is Ile.

[0064] In another embodiment of the present invention, W is -C(O)-C(CH3)2-NH-].

[0065] In another embodiment of the present invention, W is -C(O)-NH-(CH2) p -NH-], where p is 3 or 4.

[0066] In another embodiment of the present invention, W is -C(O)-NH-(CH2)4-NH-].

[0067] In another embodiment of the present invention, W is -C(O)-CH2-O-(CH2)2-O-(CH2)2-NH-].

[0068] In another embodiment of the present invention, the C-terminal amino acid is amidated as a C-terminal primary amide.

[0069] In another embodiment of the present invention, n is 16, 17, 18, 19, or 20. In a preferred embodiment, n is 18 or 20. In yet another preferred embodiment, n is 20. In another preferred embodiment, n is 16 or 18. In yet another preferred embodiment, n is 18.

[0070] In another embodiment of the present invention, Z is -C(O)-(CH2) n It is -COOH, and n is 16 or 18.

[0071] In another embodiment of the present invention, W is -C(O)-C(CH3)2-NH- and Z is -C(O)-(CH2) n It is -COOH, and n is 16.

[0072] In another embodiment of the present invention, W is -C(O)-C(CH3)2-NH- and Z is -C(O)-(CH2) n It is -COOH, and n is 18.

[0073] In another embodiment of the present invention, W is -C(O)-CH2-O-(CH2)2-O-(CH2)2-NH-] and Z is -C(O)-(CH2) n It is -COOH, and n is 16.

[0074] In another embodiment of the present invention, W is -C(O)-CH2-O-(CH2)2-O-(CH2)2-NH-] and Z is -C(O)-(CH2) n It is -COOH, and n is 18.

[0075] In another embodiment of the present invention, X5, X6, X7, X8, X9, X10, and X11 are all absent.

[0076] In another embodiment, the present invention provides a polypeptide comprising the following amino acid sequence or a pharmaceutically acceptable salt thereof, Y-X1-EGTFTSDYSI-X2-LDKIAQ-X3-AFVQWL-X4-AGGPSSGAPPPS (Sequence ID 3) In the formula, X1 is Aib, X2 is Ser(OMe) or Aib, and X4 is Ile or Glu. X3 is Lys, and in the formula, the amino(ε-amino) group of the Lys side chain is acylated at the following part: {―UWYZ In the formula, U is -C(O)-CH2-O-(CH2)2-O-(CH2)2-NH-}, where} is the attachment point to the group W. W is -C(O)-NH-(CH2)p Selected from the group consisting of -NH-], -C(O)-C(CH3)2-NH-], and -C(O)-CH2-O-(CH2)2-O-(CH2)2-NH-], where p is 3 or 4, and where ] is the attachment point with group Y. Y is -C(O)-(CH2)2-CH(COOH)NH--, and -- is the attachment point to group Z. Z is -C(O)-(CH2). n -COOH or -C(O)-(CH2) n -CH3, where n is an integer between 14 and 20. Furthermore, in the formula, the acid group of the C-terminal amino acid is either a free carboxylic acid group or is amidated as a C-terminal primary amide. However, if X2 is Aib, then W is not -C(O)-CH2-O-(CH2)2-O-(CH2)2-NH-].

[0077] In one embodiment of the present invention, X2 is Aib and X4 is Ile.

[0078] In another embodiment of the present invention, W is -C(O)-C(CH3)2-NH-].

[0079] In another embodiment of the present invention, W is -C(O)-NH-(CH2) p -NH-], where p is 3 or 4.

[0080] In another embodiment of the present invention, W is -C(O)-NH-(CH2)4-NH-].

[0081] In another embodiment of the present invention, W is -C(O)-CH2-O-(CH2)2-O-(CH2)2-NH-].

[0082] In another embodiment of the present invention, the C-terminal amino acid is amidated as a C-terminal primary amide.

[0083] In another embodiment of the present invention, n is 16, 17, 18, 19, or 20. In a preferred embodiment, n is 18 or 20. In yet another preferred embodiment, n is 20. In another preferred embodiment, n is 16 or 18. In yet another preferred embodiment, n is 18.

[0084] In another embodiment of the present invention, Z is -C(O)-(CH2) n It is -COOH, and n is 16 or 18.

[0085] In another embodiment of the present invention, W is -C(O)-NH-(CH2)4-NH- and Z is -C(O)-(CH2) n It is -COOH, and n is 18.

[0086] In another embodiment, W is -C(O)-C(CH3)2-NH- and Z is -C(O)-(CH2) n It is -COOH, and n is 16.

[0087] In another embodiment of the present invention, W is -C(O)-C(CH3)2-NH- and Z is -C(O)-(CH2) n It is -COOH, and n is 18.

[0088] In another embodiment of the present invention, X2 is Ser(OMe) and X4 is Ile.

[0089] In another embodiment of the present invention, W is -C(O)-C(CH3)2-NH- and Z is -C(O)-(CH2) n It is -COOH, and n is 16.

[0090] In another embodiment of the present invention, W is -C(O)-CH2-O-(CH2)2-O-(CH2)2-NH-] and Z is -C(O)-(CH2) n It is -COOH, and n is 16.

[0091] In another embodiment of the present invention, W is -C(O)-CH2-O-(CH2)2-O-(CH2)2-NH-] and Z is -C(O)-(CH2) n It is -COOH, and n is 18.

[0092] In another embodiment, the present invention provides a polypeptide comprising the following amino acid sequence or a pharmaceutically acceptable salt thereof, Y-Aib-EGTFTSDYSI-Aib-LDKIAQ-X3-AFVQWL-Ile-AGGPSSGAPPPS(Sequence ID 4) In the formula, X3 is Lys, and the amino(ε-amino) group of the Lys side chain is acylated at the following point: {―UWYZ In the formula, U is -C(O)-CH2-O-(CH2)2-O-(CH2)2-NH-}, where} is the attachment point to the group W. W is -C(O)-NH-(CH2) p Selected from the group consisting of -NH-] or -C(O)-C(CH3)2-NH-], where p is 3 or 4, and where ] is the attachment point to group Y. Y is -C(O)-(CH2)2-CH(COOH)NH--, and -- is the attachment point to group Z. Z is -C(O)-(CH2). n -COOH or -C(O)-(CH2) n -CH3, where n is an integer between 14 and 20. In the formula, the acid group of the C-terminal amino acid is either a free carboxylic acid group or is amidated as a C-terminal primary amide.

[0093] In another embodiment of the present invention, W is -C(O)-C(CH3)2-NH-].

[0094] In another embodiment of the present invention, W is -C(O)-NH-(CH2) p -NH-], where p is 3 or 4.

[0095] In another embodiment of the present invention, W is -C(O)-NH-(CH2)4-NH-].

[0096] In another embodiment of the present invention, the C-terminal amino acid is amidated as a C-terminal primary amide.

[0097] In another embodiment of the present invention, n is 16, 17, 18, 19, or 20. In a preferred embodiment, n is 18 or 20. In yet another preferred embodiment, n is 20. In another preferred embodiment, n is 16 or 18. In yet another preferred embodiment, n is 18.

[0098] In another embodiment of the present invention, Z is -C(O)-(CH2) n It is -COOH, and n is 16 or 18.

[0099] In another embodiment of the present invention, W is -C(O)-NH-(CH2)4-NH- and Z is -C(O)-(CH2) n It is -COOH, and n is 18.

[0100] In another embodiment of the present invention, W is -C(O)-C(CH3)2-NH- and Z is -C(O)-(CH2) n It is -COOH, and n is 16.

[0101] In another embodiment of the present invention, W is -C(O)-C(CH3)2-NH- and Z is -C(O)-(CH2) n It is -COOH, and n is 18.

[0102] In another embodiment, the present invention provides a polypeptide comprising an amino acid sequence selected from the following, or a pharmaceutically acceptable salt thereof: i)Tyr Aib Glu Gly Thr Phe Thr Ser Asp Tyr Ser Ile Aib Leu Asp Lys Ile Ala Gln X3 Ala Phe Val Gln Trp Leu Ile Ala Gly Gly Pro Ser Ser Gly Ala Pro Pro Pro Ser、 ii)Tyr Aib Glu Gly Thr Phe Thr Ser Asp Tyr Ser Ile D-Ser-(OMe) Leu Asp Lys Ile Ala Gln X3 Ala Phe Val Gln Trp Leu Ile Ala Gly Gly Pro Ser Ser Gly Ala Pro Pro Pro Ser、 iii)Tyr Aib Glu Gly Thr Phe Thr Ser Asp Tyr Ser Ile Ser(OMe) Leu Asp Lys Ile Ala Gln X3 Ala Phe Val Gln Trp Leu Ile Ala Gly Gly Pro Ser Ser Gly Ala Pro Pro Pro Ser、 iv)Tyr Aib Glu Gly Thr Phe Thr Ser Asp Tyr Ser Ile Aib Leu Asp Lys Ile Ala Gln X3 Ala Phe Val Gln Trp Leu Ile Ala Gly Gly Pro Ser Ser Gly Ala Pro Pro Pro Ser Arg、 v)Tyr Aib Glu Gly Thr Phe Thr Ser Asp Tyr Ser Ile Tyr Leu Glu Lys Ile Ala Ala Tyr Glu Phe Val Asn Trp Leu Leu Ala Gly Gly Pro Ser Ser Gly Ala Pro Pro Pro Ser X5、 vi)Tyr Aib Glu Gly Thr Phe Thr Ser Asp Tyr Ser Ile Ser(OMe) Leu Glu Lys Ile Ala Ala Gln Glu Phe Val Asn Trp Leu Leu Ala Gly Gly Pro Ser Ser Gly Ala Pro Pro Pro Ser X5, vii)Tyr D-Ser(OMe) Glu Gly Thr Phe Thr Ser Asp Tyr Ser Ile Aib Leu Asp Lys Ile Ala Gln X3 Ala Phe Val Gln Trp Leu Ile Ala Gly Gly Pro Ser Ser Gly Ala Pro Pro Pro Ser, and vii)Tyr Ser(OMe) Glu Gly Thr Phe Thr Ser Asp Tyr Ser Ile Aib Leu Asp Lys Ile Ala Gln X3 Ala Phe Val Gln Trp Leu Ile Ala Gly Gly Pro Ser Ser Ser Gly Ala Pro Pro Pro Ser In the formula, X3 and X5 have the same meaning as above. In the formula, the acid group of the C-terminal amino acid is either a free carboxylic acid group or is amidated as a C-terminal primary amide.

[0103] In another embodiment, the present invention provides a polypeptide comprising an amino acid sequence selected from the group consisting of the following: i) Tyr Aib Glu Gly Thr Phe Thr Ser Asp Tyr Ser Ile Aib Leu Asp Lys Ile Ala Gln Lys Ala Phe Val Gln Trp Leu Ile Ala Gly Gly Pro Ser Ser Gly Ala Pro Pro Pro Ser-NH2 (SEQ ID NO: 5), ii) Tyr Aib Glu Gly Thr Phe Thr Ser Asp Tyr Ser Ile D-Ser-(OMe) Leu Asp Lys Ile Ala Gln Lys Ala Phe Val Gln Trp Leu Ile Ala Gly Gly Pro Ser Ser Gly Ala Pro Pro Pro Ser-NH2 (SEQ ID NO: 9), iii) Tyr Aib Glu Gly Thr Phe Thr Ser Asp Tyr Ser Ile Ser(OMe) Leu Asp Lys Ile Ala Gln Lys Ala Phe Val Gln Trp Leu Ile Ala Gly Gly Pro Ser Ser Gly Ala Pro Pro Pro Ser-NH2 (SEQ ID NO: 10), iv) Tyr Aib Glu Gly Thr Phe Thr Ser Asp Tyr Ser Ile Aib Leu Asp Lys Ile Ala Gln Lys Ala Phe Val Gln Trp Leu Ile Ala Gly Gly Pro Ser Ser Gly Ala Pro Pro Pro Ser Arg (SEQ ID NO: 11), v) Tyr Aib Glu Gly Thr Phe Thr Ser Asp Tyr Ser Ile Tyr Leu Glu Lys Ile Ala Ala Gln Glu Phe Val Asn Trp Leu Leu Ala Gly Gly Pro Ser Ser Gly Ala Pro Pro Pro Ser Lys-NH2 (SEQ ID NO: 12), vi) Tyr Aib Glu Gly Thr Phe Thr Ser Asp Tyr Ser Ile Ser(OMe) Leu Glu Lys Ile Ala Ala Gln Glu Phe Val Asn Trp Leu Leu Ala Gly Gly Pro Ser Ser Gly Ala Pro Pro Pro Ser Lys-NH2 (SEQ ID NO: 13), vii) Tyr D-Ser(OMe) Glu Gly Thr Phe Thr Ser Asp Tyr Ser Ile Aib Leu Asp Lys Ile Ala Gln Lys Ala Phe Val Gln Trp Leu Ile Ala Gly Gly Pro Ser Ser Gly Ala Pro Pro Ser-NH2 (SEQ ID NO: 6), and viii) Tyr Ser(OMe) Glu Gly Thr Phe Thr Ser Asp Tyr Ser Ile Aib Leu Asp Lys Ile Ala Gln Lys Ala Phe Val Gln Trp Leu Ile Ala Gly Gly Pro Ser Ser Gly Ala Pro Pro Pro Ser-NH2 (SEQ ID NO: 7).

[0104] In another embodiment, the present invention provides polypeptides selected from representative compounds disclosed in Table 1 or pharmaceutically acceptable salts thereof.

[0105] In embodiments of the present invention, the following portion {―UWYZ The base U, W, Y and Z inside, or the following part {―U'- W'-Y'-Z' The groups U', W', Y', and Z' within these terms have the meanings defined herein and should not be interpreted as or confused with single-letter codes for amino acids. In the formula, the base -UWYZ and / or -U'-W'-Y'-Z' is selected from representative structures of parts A, B, C, D, and E disclosed in Table 2. The Ser(OMe) described herein is an amino acid serine, preferably an L-isomer, having the following structure due to the methylation of its hydroxyl group. [ka]

[0106] Whenever applicable, (D)Ser(OMe) refers to the D isomer of Ser(OMe).

[0107] The Tyr-(OEt) described herein is the amino acid tyrosine, preferably the L isomer, having the following structure (* indicates an attachment site to an adjacent residue) in which the hydroxyl group is ethylated. [ka]

[0108] Where applicable, (D)Tyr(OEt) always refers to the D isomer of Tyr(OEt).

[0109] Polypeptide sequences referred to herein are represented by single-letter or three-letter amino acid codes approved by IUPAC.

[0110] Unless otherwise specified, this specification is intended to cover both L and D isomers of amino acids in a sequence. However, in preferred embodiments, unless otherwise indicated, all amino acids are in the "L" form.

[0111] The "pharmaceutically acceptable salts" according to the present invention include acid addition salts formed with either organic or inorganic acids. Suitable pharmaceutically acceptable salts of the compounds of the present invention include acid addition salts that may be, for example, salts of inorganic acids such as hydrochloric acid, hydrobromic acid, or phosphoric acid, or salts of organic acids such as acetic acid, fumaric acid, methanesulfonic acid, benzoic acid, citric acid, lactic acid, fumaric acid, succinic acid, adipic acid, pimelic acid, suberic acid, azelaic acid, malic acid, tartaric acid, glutamic acid, or amino acids such as aspartic acid. The pharmaceutically acceptable acid addition salts of the present invention include, for example, salts formed by the addition of one or more acid equivalents such as monohydrochloride or dihydrochloride. Salts can be prepared by any process within the scope of the knowledge of those skilled in the art (see Berge et al., J. Pharm. Sci. 1977, 66, 1-19; and Handbook of Pharmaceutical Salts, Properties, and Use; Stahl and Wermuth, Ed.; Wiley-VCH and VHCA: Zurich, Switzerland, 2002).

[0112] Table 1 shows some representative compounds of the present invention. [Table 1] TIFF0007849310000005.tif183165 [Table 2]

[0113] In another embodiment, the present invention provides a method for treating or preventing hyperglycemia, type 2 diabetes, impaired glucose tolerance, type 1 diabetes, obesity, hypertension, hyperlipidemia, syndrome X, dyslipidemia, cognitive impairment, atherosclerosis, myocardial infarction, coronary heart disease, stroke, inflammatory bowel syndrome, dyspepsia, alcoholism, and gastric ulcers in patients, comprising administering to patients in need thereof an effective amount of the polypeptide of the present invention or a pharmaceutically acceptable salt thereof.

[0114] In another embodiment, the invention provides a method for treating type 2 diabetes in a patient, comprising administering an effective amount of the polypeptide of the present invention or a pharmaceutically acceptable salt thereof to a patient in need of such treatment.

[0115] In another embodiment, the invention provides a method for treating obesity in a patient, comprising administering to a patient in need of such treatment an effective amount of the polypeptide of the present invention or a pharmaceutically acceptable salt thereof.

[0116] In another embodiment, the invention provides a method for treating hyperlipidemia in a patient, comprising administering an effective amount of the polypeptide of the present invention or a pharmaceutically acceptable salt thereof to a patient in need of such treatment.

[0117] As used herein, the term “effective dose” or “effective amount” refers to the amount of polypeptide that is sufficient to a subject in a single or multiple doses to cure, alleviate, reduce, or partially address the clinical symptoms of a given disease or condition and its complications beyond those expected in the absence of such treatment. Thus, the result may be a reduction and / or alleviation of the signs, symptoms, or causes of the disease, or any other desirable change in the biological system. It is understood that the “therapeutic effective dose” may vary from subject to subject, depending on age, weight, the subject’s overall condition, the condition being treated, the severity of the condition being treated, and the judgment of the prescribing physician.

[0118] In one embodiment, the present invention provides a pharmaceutical composition comprising a polypeptide of the present invention or a pharmaceutically acceptable salt thereof, comprising one or more pharmaceutically acceptable carriers, diluents, or excipients.

[0119] The polypeptide of the present invention or a pharmaceutically acceptable salt thereof is preferably formulated as a pharmaceutical composition administered via parenteral routes (e.g., subcutaneous, intravenous, intraperitoneal, intramuscular, or transdermal). Such pharmaceutical compositions and processes for preparing them are well known in the art. (See, for example, Remington: The Science and 50 Practices of Pharmacy (DB Troy, Editor, 21st Edition, Lippincott, Williams & Wilkins, 2006)).

[0120] In another embodiment, the present invention provides a polypeptide of the present invention or a pharmaceutically acceptable salt thereof for use in the treatment or prevention of a patient's disease, the disease being selected from the group consisting of hyperglycemia, type 2 diabetes, impaired glucose tolerance, type 1 diabetes, obesity, hypertension, hyperlipidemia, syndrome X, dyslipidemia, cognitive impairment, atherosclerosis, myocardial infarction, coronary heart disease, stroke, inflammatory bowel syndrome, dyspepsia, alcoholism, and gastric ulcer.

[0121] In some embodiments, polypeptides or pharmaceutically acceptable salts thereof, or pharmaceutical compositions, are provided simultaneously, separately, or sequentially in combination with an effective amount of one or more additional therapeutic agents.

[0122] The present invention may be accompanied by one or more embodiments. It should be understood that the following embodiments are illustrative of the invention and are not intended to limit the claims to any particular embodiment illustrated. It should also be understood that the embodiments defined herein may be used independently or in conjunction with any other definition or embodiment defined herein. Accordingly, the present invention intends all possible combinations and permutations of various independently described embodiments. [Examples]

[0123] Instruments and analytical methods: The instrument used for characterizing and analyzing the compounds of the present invention is HPLC (Waters e2695 Alliance; Detector Waters (2489 UV / Visible)).

[0124] Equipment: HPLC: Waters e2695 Alliance; Detector: Acquity-QDa

[0125] The final compound of this disclosure was purified by a preparative HPLC procedure, as outlined below.

[0126] Preparative HPLC: WATERS 2555 Quaternary gradient module (maximum total flow rate: 300 mL / min, maximum pressure: 3000 psi) or Shimadzu LC-8A (maximum total flow rate: 150 mL, maximum pressure: 30 MPa), column: phenyl, 10 μF flow rate: 75 mL / min [Table 3]

[0127] The purity of the compounds disclosed herein was analyzed by RP-HPLC, as outlined below.

[0128] HPLC method B1: Column: YMC Pack-Phenyl (4.6mm x 150mm, 3μ) Eluent: Mobile phase A: 0.1% trifluoroacetic acid in water Mobile phase B: 0.1% trifluoroacetic acid in acetonitrile Flow rate: 1.5mL / min Detection: UV detection at 210nm Column temperature: 50℃ Execution time: 50 minutes [Table 4]

[0129] HPLC method B2: Column: Xbridge Peptide BEH C18 (4.6mm x 250mm, 3.5u) Eluent: Mobile phase A: Buffer: Acetonitrile (900:100) Mobile phase B: Buffer: Acetonitrile (300:700) Buffer solution: Potassium dihydrogen orthophosphate in water, pH adjusted to 3.0 ± 0.1 with orthophosphate. Flow rate: 1.0mL / min Detection: UV detection at 210nm Column temperature: 65℃ Sample tray temperature: 5°C Execution time: 40 minutes [Table 5]

[0130] Method B3: Column: Xbridge Peptide BEH C18 (4.6mm x 250mm, 3.5u) Eluent: Mobile phase A: Buffer: Acetonitrile (900:100) Mobile phase B: Buffer: Acetonitrile (300:700) Buffer solution: Potassium dihydrogen orthophosphate in water, pH adjusted to 3.0 ± 0.1 with orthophosphate. Flow rate: 1.0mL / min Detection: UV detection at 210nm Column temperature: 65℃ Sample tray temperature: 5°C Execution time: 65 minutes [Table 6]

[0131] Method B4: Column: Xbridge Peptide BEH C18 (4.6mm x 250mm, 3.5u) Eluent: Mobile phase A: Buffer: Acetonitrile (900:100) Mobile phase B: Buffer: Acetonitrile (300:700) Buffer solution: Potassium dihydrogen orthophosphate in water, pH adjusted to 3.0 ± 0.1 with orthophosphate. Flow rate: 0.8mL / min Detection: UV detection at 210nm Column temperature: 65℃ Sample tray temperature: 5°C Execution time: 90 minutes [Table 7]

[0132] Method B5: Column: Xbridge Peptide BEH C18 (4.6mm x 250mm, 3.5u) Eluent: Mobile phase A: Buffer: Acetonitrile (900:100) Mobile phase B: Buffer: Acetonitrile (300:700) Buffer solution: Potassium dihydrogen orthophosphate in water, pH adjusted to 3.0 ± 0.1 with orthophosphate. Flow rate: 1.0mL / min Detection: UV detection at 210nm Column temperature: 65℃ Sample tray temperature: 10℃ Execution time: 60 minutes [Table 8]

[0133] Preparation method: Example 1 Preparation of 2-[2-[2-[[2-[[(4S)-5-tert-butoxy-4-[(18-tert-butoxy-18-oxo-octadecanoyl)amino]-5-oxo-pentanoyl]amino]-2-methyl-propanoyl]amino]ethoxy]ethoxy]acetic acid (partially A-di-tert-butyl ester) [ka] Partially A-di-tert-butyl esters were prepared using solid-phase synthesis with 2-chlorotrityl chloride resin. 2-[2-(2-Fmoc-aminoethoxy)ethoxy]acetic acid was attached to 2-chlorotrityl chloride resin in the presence of DIPEA to obtain 2-[2-(2-Fmoc-aminoethoxy)ethoxy]acetic acid-2-Cl-Trt-resin. The Fmoc protecting group was removed by selective deblocking of the amino group using piperidine, and then bonded to Fmoc-Aib-OH in THF:DMAc / THF using DIPC and HOBt to obtain 2-[2-[2-[(2-Fmoc-amino-2-methyl-propanoyl)amino]ethoxy]ethoxy]acetic acid-2-Cl-Trt-resin. The Fmoc group was removed by selective deblocking using piperidine, and the free amino group was bonded to Fmoc-Glu-OtBu using HOBt and DIPC to obtain 2-[2-[2-[[2-[[(4S)-4-Fmoc-amino-5-tert-butoxy-5-oxo-pentanoyl]amino]-2-methyl-propanoyl]amino]ethoxy]ethoxy]acetic acid-2-Cl-Trt-resin. The Fmoc group of the obtained compound was selectively deblocked using piperidine, and the free amino group was then bonded to octadecanediic acid monotert-butyl ester to obtain 2-[2-[2-[[2-[[(4S)-5-tert-butoxy-4-[(18-tert-butoxy-18-oxo-octadecanoyl)amino]-5-oxo-pentanoyl]amino]-2-methyl-propanoyl]-amino]ethoxy]ethoxy]acetic acid-2-Cl-Trt-resin. The intermediate was then cleaved from the 2-Cl-Trt-resin using trifluoroethanol:DCM (1:1) to obtain the title compound (partially A-di-tert-butyl ester). (LCMS = m / z: 786.39 (M+H) + ))

[0134] Example 2 Preparation of 2-[2-[2-[[2-[[(4S)-5-tert-butoxy-4-[(20-tert-butoxy-20-oxo-icosanoyl)amino]-5-oxo-pentanoyl]amino]-2-methyl-propanoyl]amino]ethoxy]ethoxy]acetic acid (partially B-di-tert-butyl ester) [ka] 2-[2-[2-[[2-[[(4S)-4-Fmoc-amino-5-tert-butoxy-5-oxo-pentanoyl]amino]-2-methyl-propanoyl]amino]ethoxy]acetic acid-2-Cl-Trt-resin was prepared as described in Example 1 and subjected to selective deprotection using piperidine. Subsequently, the free amino group was bonded to 20-(tert-butoxy)-20-oxoeicosanoic acid to obtain 2-[2-[2-[[2-[[(4S)-5-tert-butoxy-4-[(20-tert-butoxy-20-oxo-icosanoyl)amino]-5-oxo-pentanoyl]amino]-2-methyl-propanoyl]amino]ethoxy]ethoxy]acetic acid-2-Cl-Trt-resin. Next, the intermediate was cleaved from the 2-Cl-Trt-resin using trifluoroethanol:DCM (1:1) to obtain the tile compound (partially B-di-tert-butyl ester). (LCMS=m / z:814.10(M+H) + ))

[0135] Example 3: Preparation of 2-[2-[2-[[2-[2-[2-[[5-tert-butoxy-4-[(18-tert-butoxy-18-oxo-octadecanoyl)amino]-5-oxo-pentanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetic acid (partially C-di-tert-butyl ester) [ka]

[0136] Partially C-di-tert-butyl esters were prepared using solid-phase synthesis with 2-chlorotrityl chloride resin. 2-[2-(2-Fmoc-aminoethoxy)ethoxy]acetic acid was attached to 2-chlorotrityl chloride resin in the presence of DIPEA to obtain 2-[2-(2-Fmoc-aminoethoxy)ethoxy]acetic acid-2-Cl-Trt-resin. The Fmoc protecting group was removed by selective deblocking of the amino group using piperidine, and then bonded to 2-[2-(2-Fmoc-aminoethoxy)ethoxy]acetic acid in THF using DIPC and HOBt to obtain {(Fmoc-aminoethoxy)-ethoxy}acetyl-{(-aminoethoxy)-ethoxy}acetic acid-2-Cl-Trt-resin. The Fmoc group was removed by selective deblocking using piperidine, and the free amino group was bonded to Fmoc-Glu-OtBu using HOBt and DIPC to obtain Fmoc-Glu({(amino-ethoxy)-ethoxy}acetyl-{(-amino-ethoxy)-ethoxy}acetic acid-2-Cl-Trt-resin)-OtBu. The Fmoc group of the obtained compound was selectively deblocked using piperidine, and the free amino group was then bonded to octadecanediic acid monotert-butyl ester to obtain 2-[2-[2-[[2-[2-[2-[[5-tert-butoxy-4-[(18-tert-butoxy-18-oxo-octadecanoyl)amino]-5-oxo-pentanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetic acid-2-Cl-Trt-resin. Next, using trifluoroethanol:DCM (1:1), the intermediate is cleaved from the 2-Cl-Trt resin to obtain 2-[2-[2-[[2-[2-[2-[[5-tert-butoxy-4-[(18-tert-butoxy-18-oxo-octadecanoyl)amino]-5-oxo-pentanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetic acid (partially C-di-tert-butyl ester) (LCMS=m / z:846.10(M+H + )) was obtained.

[0137] Example 4: Preparation of 2-[2-[2-[[2-[2-[2-[[5-tert-butoxy-4-[(20-tert-butoxy-20-oxo-icosanoyl)amino]-5-oxo-pentanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetic acid (Partially D-di-tert-butyl ester) [ka]

[0138] Partially D-di-tert-butyl esters were prepared using solid-phase synthesis with 2-chlorotrityl chloride resin, as schematically shown below. 2-[2-(2-Fmoc-aminoethoxy)ethoxy]acetic acid was attached to 2-chlorotrityl chloride resin in the presence of DIPEA to obtain 2-[2-(2-Fmoc-aminoethoxy)ethoxy]acetic acid-2-Cl-Trt-resin. The Fmoc protecting group was removed by selective deblocking of the amino group using piperidine, and then 2-[2-(2-Fmoc-aminoethoxy)ethoxy]acetic acid in THF was bonded using DIPC and HOBt to obtain {(Fmoc-aminoethoxy)-ethoxy}acetyl-{(-aminoethoxy)-ethoxy}acetic acid-2-Cl-Trt-resin. The Fmoc group was removed by selective deblocking using piperidine, and the free amino group was bonded to Fmoc-Glu-OtBu using HOBt and DIPC to obtain Fmoc-Glu({(amino-ethoxy)-ethoxy}-acetyl-{(-amino-ethoxy)-ethoxy}-acetic acid-2-Cl-Trt-resin)-OtBu. The Fmoc group of the obtained compound was selectively deblocked using piperidine, and the free amino group was subsequently bonded to 20-(tert-butoxy)-20-oxoicosanoic acid. [2-[2-[[2-[2-[2-[2-[[5-tert-butoxy-4-[(20-tert-butoxy-20-oxo-icosanoyl)amino]-5-oxo-pentanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetic acid]-2-Cl-Trt-resin was obtained. Then, the intermediate was cleaved from the 2-Cl-Trt-resin using trifluoroethanol:DCM (1:1), 2-[2-[2-[[2-[2-[2-[5-tert-butoxy-4-[(20-tert-butoxy-20-oxo-icosanoyl)amino]-5-oxo-pentanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetic acid (partially D-di-tert-butyl ester) (LCMS=m / z:874.15(M+H + )) was obtained.

[0139] Example 5: Preparation of 2-[2-[2-[4-[[5-tert-butoxy-4-[(20-tert-butoxy-20-oxo-icosanoyl)amino]-5-oxo-pentanoyl]amino]butylcarbamoylamino]ethoxy]ethoxy]acetic acid (partially E-di-tert-butyl ester) [ka]

[0140] Partially E-di-tert-butyl esters were prepared using solid-phase synthesis with 2-chlorotrityl chloride resin. 2-[2-(2-Fmoc-aminoethoxy)ethoxyacetic acid was attached to 2-chlorotrityl chloride resin in the presence of N,N'-di-isopropylethylamine (DIPEA) to obtain 2-[2-(2-Fmoc-aminoethoxy)ethoxy]acetic acid-2-Cl-Trt-resin. The Fmoc protecting group was removed by selective deblocking of the amino group using piperidine, and subsequently, the free amino group was activated with p-nitrophenyl chloroformate in THF and DIPEA. Then, in the presence of DIPEA, it was reacted with Fmoc-aminobutylamine hydrochloride in THF:DMAc to obtain 2-[2-[2-(4-Fmoc-aminobutylcarbamoylamino)ethoxy]ethoxy]acetic acid-2-Cl-Trt-resin. The Fmoc group is removed by selective deblocking using piperidine, and then the free amino group is bonded to Fmoc-Glu-OtBu using 1-hydroxybenzotriazole (HOBt) and N,N'-di-isopropylcarbodiimide (DIPC) to form 2-[2-[2-[4-[[(4S)-4-Fmoc-amino-5-tert-butoxy-5-oxo-pentanoyl]amino]butylcarbamoylamino]ethoxy]ethoxy ]acetic acid-2-Cl-Trt resin was obtained, which was selectively deblocked using piperidine and then bonded with 20-(tert-butoxy)-20-oxoeicosanoic acid to obtain the intermediate 2-[2-[2-[4-[[5-tert-butoxy-4-[(20-tert-butoxy-20-oxo-icosanoyl)amino]-5-oxo-pentanoyl]amino]butylcarbamoylamino]ethoxy]ethoxy]acetic acid-2-Cl-Trt resin. Next, using trifluoroethanol:DCM (1:1), the intermediate is cleaved from the 2-Cl-Trt resin to obtain 2-[2-[2-[4-[[5-tert-butoxy-4-[(20-tert-butoxy-20-oxo-icosanoyl)amino]-5-oxo-pentanoyl]amino]butylcarbamoylamino]ethoxy]ethoxy]acetic acid (LCMS=m / z:843.14(M+H +)) was obtained. (Partially E-di-tert-butyl ester).

[0141] Example 6: Preparation of Compound 1 The parent peptide was synthesized by a solid-phase method. The starting resin used for synthesis was Fmoc-Rink amide resin. The Fmoc-protecting amino groups of the Rink amide resin were selectively deblocked using piperidine, followed by the bonding of Fmoc-Ser(tBu)-OH to the Rink amide resin. Bonding was performed using DIPC-HOBt to obtain Fmoc-Ser(tBu)-Rink amide resin, completing one cycle. Unbound amino groups at each amino acid bond were terminated / capped using acetic anhydride and DIPEA / pyridine. Selective deblocking of the amino groups of the Fmoc-Ser(tBu)-Rink amide resin using piperidine, followed by bonding with Fmoc-Pro-OH using HOBt and DIPC, resulted in the obtaining of Fmoc-Pro-Ser(tBu)-rink amide resin. This completed the second cycle. Unbound amino groups at each amino acid bond were terminated using acetic anhydride and DIPEA / pyridine.

[0142] The three steps described above—selective capping, deblocking, and linking of adjacent amino acid residues in the sequence with the Fmoc-protected amino acid group—were repeated for the remaining 36 amino acid residues, with the final link being with a Boc-protected amino acid (i.e., Boc-Tyr(tBu)-OH). Selective deblocking, i.e., capping of unbound amino groups, was performed using acetic anhydride and DIPEA / pyridine, deprotection of the Fmoc / Boc group was performed using piperidine, and linking with adjacent Fmoc and / or Boc-protected amino acids was performed using HOBt / DIPC. The side chains of Fmoc / Boc protected amino acids are protected at right angles; for example, the hydroxyl group of serine, tyrosine, or threonine are protected with a tert-butyl (-tBu) group, the amino group of lysine is protected with a tert-butyloxycarbonyl (-Boc) and (4,4-dimethyl-2,6-dioxocyclohex-1-ylidene)-3-methylbutyl (IVDde) group, respectively, the carboxylic acid group of aspartic acid or glutamic acid is protected with a tBu group, and the amide group of glutamine is protected with a trityl (-Trt) group. The three steps described above—selective capping, deblocking, and subsequent bonding with the adjacent Fmoc-protecting amino acid—were performed, with Boc-Tyr(tBu)-OH also used last, to obtain the Boc-Tyr(tBu)-Aib-Glu(OtBu)-Gly-Thr(tBu)-Phe-Thr(tBu)-Ser(tBu)-Asp(OtBu)-Tyr(tBu)-Ser(tBu)-Ile-Aib-Leu-Asp(OtBu)-Lys(Boc)-Ile-Ala-Gln(Trt)-Lys(IVDde)-Ala-Phe-Val-Gln(Trt)-Trp-Leu-Ile-Ala-Gly-Gly-Pro-Ser(tBu)-Ser(tBu)-Gly-Ala-Pro-Pro-Pro-Ser(tBu)-Rink amide resin.

[0143] Deprotection of the IVDde group of a peptide resin using hydrazine hydrate, followed by bonding of a partial A-di-tert butyl ester, was performed using DIPC-HOBt to obtain a protected compound 1 resin.

[0144] Boc-Tyr(tBu)-Aib-Glu(OtBu)-Gly-Thr(tBu)-Phe-Thr(tBu)-Ser(tBu)-Asp(OtBu)-Tyr(tBu)-Ser(tBu)-Ile-Aib-Leu-Asp(OtBu)-Lys(Boc)-Ile-Ala-Gln(Trt)-Lys(NH-partial A di-tert-butyl ester)-Ala-Phe-Val-Gln(Trt)-Trp-Leu-Ile-Ala-Gly-Gly-Pro-Ser(tBu)-Ser(tBu)-Gly-Ala-Pro-Pro-Pro-Ser(tBu)-Rink amide resin. Cleavage and deprotection using trifluoroacetic acid with ethane-1,2-dithiol and triisopropylsilane, followed by purification by preparative HPLC, yielded compound 1. The HPLC purity of compound 1 was evaluated by method B2. Mass (LCMS): m / z = 1182.41 (MH4 4+ ), calculated mass = 4725.61, HPLC purity: 97.77% (Method B2), RT = 19.9 min

[0145] Example 7: Synthesis of Compound 2: Compound 2 was prepared by a solid-phase method following a similar process to that described in Example 6, except that a partial β-di-tert-butyl ester was bonded to the peptide resin. Compound 2 was then obtained by cleavage, deprotection, and preparative purification using HPLC. The HPLC purity of Compound 2 was evaluated by Method B2.

[0146] Mass (LCMS): m / z=1189.36(MH4 4+ ), calculated mass = 4753.41; HPLC purity: 94.50% (Method B2), RT = 22.1 min

[0147] Example 8: Synthesis of Compound 3: The parent peptide was synthesized by a solid-phase method. The starting resin used for synthesis was Fmoc-Rink amide resin. The Fmoc-protecting amino groups of the rink amide resin were selectively deblocked using piperidine, followed by the bonding of Fmoc-Ser(tBu)-OH to the Rink amide resin. Bonding was performed using DIPC-HOBt to obtain Fmoc-Ser(tBu)-Rink amide resin, completing one cycle. Unbonded amino groups at each amino acid bond were terminated / capped using acetic anhydride and diisopropylethylamine / pyridine. Selective deblocking of the amino groups of the Fmoc-Ser(tBu)-Rink amide resin using piperidine, followed by bonding with Fmoc-Pro-OH using HOBt and DIPC, resulted in the obtaining of Fmoc-Pro-Ser(tBu)-rink amide resin. This completed the second cycle. Unbonded amino groups at each amino acid bond were terminated using acetic anhydride and diisopropylethylamine / pyridine.

[0148] The three steps described above—selective capping, deblocking, and binding of the Fmoc-protected amino group of an adjacent amino acid residue to the resin—were repeated for the remaining 37 amino acid residues. Selective deblocking, i.e., capping of unbound amino groups, was performed using acetic anhydride and diisopropylethylamine / pyridine, deprotection of the Fmoc group was performed using piperidine, and binding to the adjacent Fmoc-protected amino acid was performed using HOBt / DIPC. The side chains of Fmoc-protected amino acids are protected at right angles; for example, the hydroxyl group of serine is protected with a tert-butyl (-tBu) group and a methyl (OMe) group, tyrosine or threonine is protected with a tert-butyl (-tBu) group, the amino group of lysine is protected with a tert-butyloxycarbonyl (-Boc) group and a (4,4-dimethyl-2,6-dioxocyclohex-1-ylidene)-3-methylbutyl (IVDde) group, respectively, the carboxylic acid group of aspartic acid or glutamic acid is protected with a tBu group, and the amide group of glutamine is protected with a trityl (-Trt) group. The three steps described above—selective capping, deblocking, and subsequent binding with adjacent Fmoc-protecting amino acids—were performed to obtain the Fmoc-Tyr(tBu)-(D)Ser(OMe)-Glu(OtBu)-Gly-Thr(tBu)-Phe-Thr(tBu)-Ser(tBu)-Asp(OtBu)-Tyr(tBu)-Ser(tBu)-Ile-Aib-Leu-Asp(OtBu)-Lys(Boc)-Ile-Ala-Gln(Trt)-Lys(IVDde)-Ala-Phe-Val-Gln(Trt)-Trp-Leu-Ile-Ala-Gly-Gly-Pro-Ser(tBu)-Ser(tBu)-Gly-Ala-Pro-Pro-Pro-Ser(tBu)-Rink amide resin.

[0149] Following deblocking of the Fmoc-Tyr(tBu)-(D)Ser(OMe)-Glu(OtBu)-Gly-Thr(tBu)-Phe-Thr(tBu)-Ser(tBu)-Asp(OtBu)-Tyr(tBu)-Ser(tBu)-Ile-Aib-Leu-Asp(OtBu)-Lys(Boc)-Ile-Ala-Gln(Trt)-Lys(IVDde)-Ala-Phe-Val-Gln(Trt)-Trp-Leu-Ile-Ala-Gly-Gly-Pro-Ser(tBu)-Ser(tBu)-Gly-Ala-Pro-Pro-Pro-Ser(tBu)-Rink amide resin with piperidine, Boc By protecting the peptide resin using anhydrous substances with Boc, we obtained a Boc-Tyr(tBu)-(D)Ser(OMe)-Glu(otBu)-Gly-Thr(tBu)-Phe-Thr(tBu)-Ser(tBu)-Asp(OtBu)-Tyr(tBu)-Ser(tBu)-Ile-Aib-Leu-Asp(OtBu)-Lys(Boc)-Ile-Ala-Gln(Trt)-Lys(IVDde)-Ala-Phe-Val-Gln(Trt)-Trp-Leu-Ile-Ala-Gly-Gly-Pro-Ser(tBu)-Ser(tBu)-Gly-Ala-Pro-Pro-Pro-Ser(tBu)-Rink amide resin. Following the deprotection of the IVDde group of the peptide resin using hydrazine hydrate, the partial β-di-tert butyl ester was bonded using diisopropylcarbodiimide and N-hydroxybenzotriazole (DIPC-HOBt) as coupling reagents to obtain compound 3 resin in their presence.

[0150] Boc-Tyr(tBu)-(D)Ser(OMe)-Glu(OtBu)-Gly-Thr(tBu)-Phe-Thr(tBu)-Ser(tBu)-Asp(Ot Bu)-Tyr(tBu)-Ser(tBu)-Ile-Aib-Leu-Asp(OtBu)-Lys(Boc)-Ile-Ala-Gln(Trt)-Lys(NH -Partially B-di-tert butyl ester)-Ala-Phe-Val-Gln(Trt)-Trp-Leu-Ile-Ala-Gly-Gly-Pro-Ser(tBu)-Ser(tBu)-Gly-Ala-Pro-Pro-Pro-Ser(tBu)-Rink amide resin. Cleavage and deprotection using trifluoroacetic acid with ethane-1,2-dithiol and triisopropylsilane, followed by purification by preparative HPLC, yielded compound 3. The HPLC purity of compound 3 was evaluated by method B2.

[0151] Mass (LCMS): m / z=1193.70(MH4 4+ ), calculated mass = 4770.77, HPLC purity: 91.96% (Method B2), RT = 29.0 min

[0152] Example 9: Synthesis of Compound 4: Compound 4 was prepared by a solid-phase method according to a similar process described in Example 8, but in Compound 4, Fmoc-Ser(OMe)-OH was used at position 2 instead of Fmoc-D-Ser(OMe)-OH, resulting in Boc-Tyr(tBu)-Ser(OMe)-Glu(OtBu)-Gly-Thr(tBu)-Phe-Thr(tBu)-Ser(tBu)-Asp(OtBu)-Tyr(tBu) -Ser(tBu)-Ile-Aib-Leu-Asp(OtBu)-Lys(Boc)-Ile-Ala-Gln(Trt)-Lys(IVDde)-Ala-Phe-Val-Gln(Trt)-Trp-Leu-Ile-Ala-Gly-Gly-Pro-Ser(tBu)-Ser(tBu)-Gly-Ala-Pro-Pro-Pro-Ser(tBu)-Rink amide resin was obtained. Next, compound 4 was obtained by bonding with a partial B-di-tert butyl ester, followed by cleavage, deprotection, and preparative purification using HPLC. The HPLC purity of compound 4 was evaluated by method B2.

[0153] Mass (LCMS): m / z=1193.68(MH4 4+ ), calculated mass = 4770.69, HPLC purity: 95.52% (Method B2), RT = 26.2 min

[0154] Example 10: Synthesis of Compound 5: Compound 5 was prepared by a solid-phase method according to a similar process described in Example 8, but in Compound 5, Fmoc-(D)-Tyr(OEt)-OH was used at position 1 instead of Fmoc-Tyr(tBu)-OH, and Fmoc-Aib-OH was used at position 2 instead of Fmoc-D-Ser(OMe)-OH, and Boc-(D)-Tyr(OEt)-Aib-Glu(OtBu)-Gly-Thr(tBu)-Phe-Thr(tBu)-Ser (tBu)-Asp(OtBu)-Tyr(tBu)-Ser(tBu)-Ile-Aib-Leu-Asp(OtBu)-Lys(Boc)-Ile-Ala-Gln(Trt)-Lys(IVDde)-Ala-Phe- Val-Gln(Trt)-Trp-Leu-Ile-Ala-Gly-Gly-Pro-Ser(tBu)-Ser(tBu)-Gly-Ala-Pro-Pro-Pro-Ser(tBu)-Rink amide resin was obtained.

[0155] Next, compound 5 was obtained by bonding with a partial β-di-tertbutyl ester, followed by cleavage, deprotection, and preparative purification using HPLC. The HPLC purity of compound 5 was evaluated by method B3.

[0156] Mass (LCMS): m / z=1196.34(MH4 4+ ), calculated mass = 4781.33, HPLC purity: 93.86% (Method B3), RT = 38.8 min

[0157] Example 11: Synthesis of Compound 6: Compound 6 was prepared by a solid-phase method according to a similar process described in Example 8, but in Compound 6, Fmoc-(D)Ser(OMe)-OH was used at position 13 instead of Fmoc-Aib-OH, and Fmoc-Aib-OH was used at position 2 instead of Fmoc-D-Ser(OMe)-OH, resulting in Boc-Tyr(tBu)-Aib-Glu(OtBu)-Gly-Thr(tBu)-Phe-Thr(tBu)-Ser(tBu)-Asp (OtBu)-Tyr(tBu)-Ser(tBu)-Ile-(D)Ser(OMe)-Leu-Asp(OtBu)-Lys(Boc)-Ile-Ala-Gln(Trt)-Lys(IVDde)-Ala-Phe- Val-Gln(Trt)-Trp-Leu-Ile-Ala-Gly-Gly-Pro-Ser(tBu)-Ser(tBu)-Gly-Ala-Pro-Pro-Pro-Ser(tBu)-Rink amide resin was obtained.

[0158] Next, compound 6 was obtained by bonding with a partial β-di-tertbutyl ester, followed by cleavage, deprotection, and preparative purification using HPLC. The HPLC purity of compound 6 was evaluated by method B2.

[0159] Mass (LCMS): m / z=1191.03(MH4 4- ), Calculated mass: 4768.15; HPLC purity: 94.74% (Method B2), RT = 27.1 min

[0160] Example 12: Synthesis of Compound 7: Compound 7 was prepared by a solid-phase method according to a similar process described in Example 8, but in Compound 7, Fmoc-Ser(OMe)-OH was used at position 13 instead of Fmoc-Aib-OH, and Fmoc-Aib-OH was used at position 2 instead of Fmoc-D-Ser(OMe)-OH, resulting in Boc-Tyr(tBu)-Aib-Glu(OtBu)-Gly-Thr(tBu)-Phe-Thr(tBu)-Ser(tBu)-Asp (OtBu)-Tyr(tBu)-Ser(tBu)-Ile-Ser(OMe)-Leu-Asp(OtBu)-Lys(Boc)-Ile-Ala-Gln(Trt)-Lys(IVDde)-Ala-Phe-Va l-Gln(Trt)-Trp-Leu-Ile-Ala-Gly-Gly-Pro-Ser(tBu)-Ser(tBu)-Gly-Ala-Pro-Pro-Pro-Ser(tBu)-Rink amide resin was obtained.

[0161] Next, compound 7 was obtained by bonding with a partial β-di-tertbutyl ester, followed by cleavage, deprotection, and preparative purification using HPLC. The HPLC purity of compound 7 was evaluated by method B2.

[0162] Mass (LCMS): m / z=1193.67(MH4 4+ ), Calculated mass = 4770.65, HPLC purity: 95.4% (Method B2), RT = 26.4 min

[0163] Example 13: Compound 8: Compound 8 was prepared by a solid-phase method according to a similar process described in Example 8, but in Compound 8, Fmoc-Aib-OH was used at position 2 instead of Fmoc-D-Ser(OMe)-OH to obtain Boc-Tyr(tBu)-Aib-Glu(OtBu)-Gly-Thr(tBu)-Phe-Thr(tBu)-Ser(tBu)-Asp(OtBu)-Tyr(tBu)-Ser(tBu)-Ile-Aib-Leu-Asp(OtBu)-Lys(Boc)-Ile-Ala-Gln(Trt)-Lys(IVDde)-Ala-Phe-Val-Gln(Trt)-Trp-Leu-Ile-Ala-Gly-Gly-Pro-Ser(tBu)-Ser(tBu)-Gly-Ala-Pro-Pro-Pro-Ser(tBu)-Rink amide resin.

[0164] Next, compound 8 was obtained by bonding with a partial E-di-tertbutyl ester, followed by cleavage, deprotection, and preparative purification using HPLC, resulting in compound formation. The HPLC purity of compound 8 was evaluated by method B4.

[0165] Mass (LCMS): m / z=1196.55(MH4 4+ ), Calculated mass = 4782.168, HPLC purity: 97.37% (Method B4), RT = 25.6 min

[0166] Example 14: Synthesis of Compound 9: Compound 9 was prepared by a solid-phase method according to a similar process described in Example 8, but in compound 9, Fmoc-Ser(OMe)-OH was used at position 13 instead of Fmoc-Aib-OH, and Fmoc-Aib-OH was used at position 2 instead of Fmoc-D-Ser(OMe)-OH, resulting in Boc-Tyr(tBu)-Aib-Glu(OtBu)-Gly-Thr(tBu)-Phe-Thr(tBu)-Ser(tBu)-Asp (OtBu)-Tyr(tBu)-Ser(tBu)-Ile-Ser(OMe)-Leu-Asp(OtBu)-Lys(Boc)-Ile-Ala-Gln(Trt)-Lys(IVDde)-Ala-Phe-Va l-Gln(Trt)-Trp-Leu-Ile-Ala-Gly-Gly-Pro-Ser(tBu)-Ser(tBu)-Gly-Ala-Pro-Pro-Pro-Ser(tBu)-Rink amide resin was obtained.

[0167] Next, compound 9 was obtained by bonding with a partial C-di-tertbutyl ester, followed by cleavage, deprotection, and preparative purification using HPLC. The HPLC purity of compound 9 was evaluated by method B2.

[0168] Mass (LCMS): m / z=1201.7(MH4 4+ ), Calculated mass = 4802.8, HPLC purity: 97.30% (Method B2), RT = 15.3 min

[0169] Example 15: Synthesis of Compound 10: Compound 10 was prepared by a solid-phase method according to a similar process described in Example 8, but in compound 10, Fmoc-Ser(OMe)-OH was used at position 13 instead of Fmoc-Aib-OH, and Fmoc-Aib-OH was used at position 2 instead of Fmoc-D-Ser(OMe)-OH, and Boc-Tyr(tBu)-Aib-Glu(OtBu)-Gly-Thr(tBu)-Phe-Thr(tBu)-Ser(tBu)-Asp(OtBu)- Tyr(tBu)-Ser(tBu)-Ile-Ser(OMe)-Leu-Asp(OtBu)-Lys(Boc)-Ile-Ala-Gln(Trt)-Lys(IVDde)-Ala-Phe-Val-G ln(Trt)-Trp-Leu-Ile-Ala-Gly-Gly-Pro-Ser(tBu)-Ser(tBu)-Gly-Ala-Pro-Pro-Pro-Ser(tBu)-Rink amide resin was obtained.

[0170] Next, compound 10 was obtained by bonding with a partial D-di-tertbutyl ester, followed by cleavage, deprotection, and preparative purification using HPLC. The HPLC purity of compound 10 was evaluated by method B2.

[0171] Mass (LCMS): m / z=1610.78(MH3 3+ ), Calculated mass = 4829.316, HPLC purity: 93.41% (Method B2), RT = 20.3 min

[0172] Example 16: Synthesis of Compound 11: The parent peptide was synthesized by solid-phase method. The starting resin used in the synthesis was Wang resin. Fmoc-protected Arg(pbf) was used for bonding with the Wang resin. Bonding was carried out in the presence of 4-dimethylaminopyridine (DMAP) using diisopropylcarbodiimide and N-hydroxybenzotriazole (DIC-HOBt) as bonding reagents to obtain Fmoc-Arg(pbf)-Wang resin. Selective deblocking of the amino groups of Fmoc-Arg(pbf)-Wang resin using piperidine, followed by bonding with Fmoc-Ser(tBu)-OH using HOBt / DIPC, obtained Fmoc-Ser(tBu)-Arg(pbf)-Wang resin. This completed one cycle. Unbound amino groups at each amino acid bond were terminated using acetic anhydride and diisopropylethylamine / pyridine.

[0173] The three steps described above—selective deblocking of Fmoc protection of amino acids attached to the resin, binding of adjacent amino acid residues in the sequence to the Fmoc-protected amino group, and capping—were repeated for the remaining 38 amino acid residues to protect the side chains of the Fmoc-protected amino acids at right angles. For example, the hydroxyl group of serine, tyrosine, or threonine was protected with a tert-butyl (-tBu) group; the amino group of lysine was protected with a tert-butyloxycarbonyl (-Boc) and (4,4-dimethyl-2,6-dioxocyclohex-1-ylidene)-3-methylbutyl (IVDde) group, respectively; the carboxylic acid group of aspartic acid or glutamic acid was protected with a -tBu group, respectively; the amide group of glutamine was protected with a trityl (-Trt) group; and the side chain of arginine was protected with a pbf group. The three steps described above—selective capping, deblocking, and subsequent binding with adjacent Fmoc-protecting amino acids—were performed to obtain Fmoc-Tyr(tBu)-Aib-Glu(OtBu)-Gly-Thr(tBu)-Phe-Thr(tBu)-Ser(tBu)-Asp(OtBu)-Tyr(tBu)-Ser(tBu)-Ile-Aib-Leu-Asp(OtBu)-Lys(Boc)-Ile-Ala-Gln(Trt)-Lys(IVDde)-Ala-Phe-Val-Gln(Trt)-Trp-Leu-Ile-Ala-Gly-Gly-Pro-Ser(tBu)-Ser(tBu)-Gly-Ala-Pro-Pro-Pro-Ser(tBu)-Arg(pbf)-Wang resin.

[0174] Following deblocking of Fmoc-Tyr(tBu)-Aib-Glu(OtBu)-Gly-Thr(tBu)-Phe-Thr(tBu)-Ser(tBu)-Asp(OtBu)-Tyr(tBu)-Ser(tBu)-Ile-Aib-Leu-Asp(OtBu)-Lys(Boc)-Ile-Ala-Gln(Trt)-Lys(IVDde)-Ala-Phe-Val-Gln(Trt)-Trp-Leu-Ile-Ala-Gly-Gly-Pro-Ser(tBu)-Ser(tBu)-Gly-Ala-Pro-Pro-Pro-Ser(tBu)-Arg(pbf)-Wang resin with piperidine, Boc By protecting the peptide resin using anhydrous peptides with Boc, we obtained Boc-Tyr(tBu)-Aib-Glu(OtBu)-Gly-Thr(tBu)-Phe-Thr(tBu)-Ser(tBu)-Asp(OtBu)-Tyr(tBu)-Ser(tBu)-Ile-Aib-Leu-Asp(OtBu)-Lys(Boc)-Ile-Ala-Gln(Trt)-Lys(IVDde)-Ala-Phe-Val-Gln(Trt)-Trp-Leu-Ile-Ala-Gly-Gly-Pro-Ser(tBu)-Ser(tBu)-Gly-Ala-Pro-Pro-Pro-Ser(tBu)-Arg(pbf)-Wang resin. The IVDde group of the peptide resin was deprotected using hydrazine hydrate, and then coupled with a partial B-di-tert butyl ester using diisopropylcarbodiimide and N-hydroxybenzotriazole (DIPC-HOBt) as coupling reagents to obtain the intermediate protection compound 11 resin. Cleavage and deprotection from the resin using trifluoroacetic acid containing ethane-1,2-dithiol and triisopropylsilane, followed by purification by preparative HPLC, yielded compound 11.

[0175] The HPLC purity of compound 11 was evaluated using method B2.

[0176] Mass (LCMS): m / z=1228.8(MH4 4+ ), calculated mass = 4911.17, HPLC purity: 98.22% (Method B2), RT = 23.3 min

[0177] Example 17: Synthesis of Compound 12: Compound 12 was prepared by a solid-phase method according to a similar process to that described in Example 16, except that a partial C-di-tert-butyl ester was bonded to the peptide resin. Compound 12 was then obtained by cleavage, deprotection, and preparative purification using HPLC. The HPLC purity of Compound 12 was evaluated by Method B2.

[0178] Mass (LCMS): m / z=1236.56(MH4 4+ ), calculated mass = 4942.21, HPLC purity: 97.2% (Method B2), RT = 11.703 min

[0179] Example 18: Synthesis of Compound 13: Compound 13 was prepared by a solid-phase method following a similar process to that described in Example 12, except that a partial A-di-tert-butyl ester was bonded to the peptide resin. Compound 13 was then obtained by cleavage, deprotection, and preparative purification using HPLC. The HPLC purity of Compound 13 was evaluated by Method B2.

[0180] Mass (LCMS): m / z=1579.52(MH3 3- ), Calculated mass = 4741.548, HPLC purity: 96.5% (Method B2), RT = 14.76 min

[0181] Example 19: Synthesis of Compound 14: The parent peptide was synthesized by a solid-phase method. The starting resin used for synthesis was Fmoc-Rink amide resin. The Fmoc-protecting amino group of the Rink amide resin was selectively deblocked using piperidine, followed by the bonding of Fmoc-Lys(IVDde)-OH to the Rink amide resin. Bonding was performed using DIPC-HOBt to obtain Fmoc-Lys(IVDde)-Rink amide resin, completing one cycle. Acetic anhydride and diisopropylethylamine / pyridine were used to terminate / cap the unbonded amino group at the end of each amino acid bond. Selective deblocking of the Fmoc amino group of the Fmoc-Lys(IVDde)-Rink amide resin using piperidine, followed by bonding with the second amino acid using HOBt and DIPC, yielded Fmoc-Ser(tBu)-Lys(IVDde)-rink amide resin. This completed the second cycle. As mentioned earlier, acetic anhydride and diisopropylethylamine / pyridine were used to terminate the unbound amino groups in each amino acid bond.

[0182] The above three steps, namely, deblocking of the Fmoc protection of the amino acids attached to the resin, coupling with the Fmoc-protected amino group of the adjacent amino acid residue in the sequence, and selective capping, were repeated for the remaining 38 amino acid residues. The side chains of the Fmoc-protected amino acids used were protected at right angles. For example, the hydroxyl group of serine, tyrosine or threonine was protected with a tert-butyl (-tBu) group, the amino group of lysine was protected with tert-butyloxycarbonyl (-Boc) and (4,4-dimethyl-2,6-dioxocyclohex-1-ylidene)-3-methylbutyl (IVDde) groups, respectively, the carboxylic acid group of aspartic acid or glutamic acid was protected with a tBu group, and the amide groups of glutamine and asparagine were protected with a trityl (-Trt) group. The above three steps, namely, selective capping, deblocking, and subsequent coupling with the adjacent Fmoc-protected amino acid were carried out to obtain Fmoc-Tyr(tBu)-Aib-Glu(OtBu)-Gly-Thr(tBu)-Phe-Thr(tBu)-Ser(tBu)-Asp(OtBu)-Tyr(tBu)-Ser(tBu)-Ile-Tyr(tBu)-Leu-Glu(OtBu)-Lys(Boc)-Ile-Ala-Ala-Gln(Trt)-Glu(OtBu)-Phe-Val-Asn(Trt)-Trp-Leu-Leu-Ala-Gly-Gly-Pro-Ser(tBu)-Ser(tBu)-Gly-Ala-Pro-Pro-Pro-Ser(tBu)-Lys(IVDde)-Rink amide resin.

[0183] Following deblocking of the Fmoc-Tyr(tBu)-Aib-Glu(OtBu)-Gly-Thr(tBu)-Phe-Thr(tBu)-Ser(tBu)-Asp(OtBu)-Tyr(tBu)-Ser(tBu)-Ile-Tyr(tBu)-Leu-Glu(OtBu)-Lys(Boc)-Ile-Ala-Ala-Gln(Trt)-Glu(OtBu)-Phe-Val-Asn(Trt)-Trp-Leu-Leu-Ala-Gly-Gly-Pro-Ser(tBu)-Ser(tBu)-Gly-Ala-Pro-Pro-Pro-Ser(tBu)-Lys(IVDde)-Rink amide resin with piperidine, Boc By protecting the peptide resin using anhydrous substances with Boc, we obtained a Boc-Tyr(tBu)-Aib-Glu(OtBu)-Gly-Thr(tBu)-Phe-Thr(tBu)-Ser(tBu)-Asp(OtBu)-Tyr(tBu)-Ser(tBu)-Ile-Tyr(tBu)-Leu-Glu(OtBu)-Lys(Boc)-Ile-Ala-Ala-Gln(Trt)-Glu(OtBu)-Phe-Val-Asn(Trt)-Trp-Leu-Leu-Ala-Gly-Gly-Pro-Ser(tBu)-Ser(tBu)-Gly-Ala-Pro-Pro-Pro-Ser(tBu)-Lys(IVDde)-Rink amide resin. Following the deprotection of the IVDde group of the peptide resin using hydrazine hydrate, a partial B-di-tert butyl ester was bonded using diisopropylcarbodiimide and N-hydroxybenzotriazole (DIPC-HOBt) as coupling reagents to obtain compound 14 resin in their presence.

[0184] Compound 14 was obtained by cleavage and deprotection of Boc-Tyr(tBu)-Aib-Glu(OtBu)-Gly-Thr(tBu)-Phe-Thr(tBu)-Ser(tBu)-Asp(OtBu)-Tyr(tBu)-Ser(tBu)-Ile-Tyr(tBu)-Leu-Glu(OtBu)-Lys(Boc)-Ile-Ala-Ala-Gln(Trt)-Glu(OtBu)-Phe-Val-Asn(Trt)-Trp-Leu-Leu-Ala-Gly-Gly-Pro-Ser(tBu)-Ser(tBu)-Gly-Ala-Pro-Pro-Pro-Ser(tBu)-Lys(NH part B-di-tert-butyl ester)-Rink amide resin using trifluoroacetic acid with ethanedithiol and triisopropylsilane, followed by purification by preparative HPLC. The HPLC purity of compound 14 was evaluated by method B5.

[0185] Mass (LCMS): m / z = 993.06 (MH5 5+ ), calculated mass = 4960.26, HPLC purity: 95.8% (method B5), RT = 28.308 min

[0186] Example 20: Synthesis of compound 15: Compound 15 was prepared by a solid-phase method according to a similar process described in Example 19, but in compound 15, Fmoc-Ser(OMe)-OH was used at position 13 instead of Fmoc-Tyr(tBu) to obtain Fmoc-Tyr(tBu)-Aib-Glu(OtBu)-Gly-Thr(tBu)-Phe-Thr(tBu)-Ser(tBu)-Asp(OtBu)-Tyr(tBu)-Ser(tBu)-Ile-Ser(OMe)-Leu-Glu(OtBu)-Lys(Boc)-Ile-Ala-Ala-Gln(Trt)-Glu(OtBu)-Phe-Val-Asn(Trt)-Trp-Leu-Leu-Ala-Gly-Gly-Pro-Ser(tBu)-Ser(tBu)-Gly-Ala-Pro-Pro-Pro-Ser(tBu)-Lys(IVDde)-Rink amide resin.

[0187] Next, compound 15 was obtained by bonding with a partial β-di-tertbutyl ester, followed by cleavage, deprotection, and preparative purification using HPLC. The HPLC purity of compound 15 was evaluated by method B4.

[0188] Mass (LCMS): m / z=980.77(MH5 5+ ), calculated mass = 4898.8, HPLC purity: 94% (Method B4), RT = 41.5 min

[0189] Example 21: Synthesis of Compound 16: Compound 16 was prepared by a solid-phase method according to a similar process to that described in Example 16, except that a partial D-di-tert-butyl ester was bonded to the peptide resin. Compound 16 was then obtained by cleavage, deprotection, and preparative purification using HPLC. The HPLC purity of Compound 16 was evaluated by Method B2.

[0190] Mass (LCMS): m / z=1243.60(MH4 4+ ), calculated mass = 4970.37, HPLC purity: 97.5% (Method B2), RT = 19.183 mins

[0191] Biological research Example 22: Reduction of HbA1c in db / db2 diabetic mice after chronic treatment The effects of compound 2 on %HbA1c, insulin, triglyceride levels, food intake, and body weight were studied in mice. This study was conducted using a type 2 diabetic mouse (db / db) model. Animals were divided into six treatment groups (n=8 per group): a diabetes control group, compound 2 (4.5 nM / kg, 9 nM / kg, and 18 nM / kg), and tilzepatide (90 nM / kg and 180 nM / kg) treatment groups. All treatments were administered subcutaneously in 10 doses every three days (q3d*10). %HbA1c, insulin, and triglyceride levels were measured on days 0, 14, and 28. Cumulative food intake from days 0 to 28 and the percentage change in body weight compared to day 0 were calculated on day 28. Results are provided in Table 3. Table 9

[0192] As is evident from the results, compound 2 at doses of 4.5, 9, and 18 nM / kg showed a statistically significant change in HbA1c compared to diabetic controls on both day 14 and day 28. The reduction in HbA1c with compound 2 exceeded the change shown by tilzepatide at a dose of 90 nM / kg. Compound 2 showed a similar effect on insulin levels, with a statistically significant increase in insulin levels at a dose of 9 nM / kg compared to the diabetic control group on day 14. The increase in insulin levels was maintained even on day 28. By comparison, tilzepatide at a 10-fold higher dose (90 nM / kg) showed a comparable effect on insulin levels. Furthermore, it was surprising to find that the effect on insulin levels shown by compound 2 at a dose of 18 nM / kg was equivalent to the effect shown by tilzepatide at a dose of 180 nM / kg. Insulin levels with compound 2 at 18 nM were maintained at similar levels on both day 14 and day 28, although with tirzepatide treatment, insulin levels on day 28 tended to be slightly lower than on day 14. Compound 2 at doses of 4.5, 9, and 18 nM / kg showed a statistically significant reduction in body weight compared to the diabetic control group on day 28. Surprisingly, the effect of compound 2 on body weight reduction was superior to the effect shown by tirzepatide at a dose of 180 nM / kg (20 times the dose). Compound 2 at the test doses (4.5, 9, and 18 nM / kg) also showed a statistically significant reduction in cumulative food consumption compared to the diabetic control group during the study. Surprisingly, the effect of compound 2 on food consumption was comparable to the effect shown by tirzepatide at a dose 10 times that of compound 2. Similarly, compound 2 at doses of 4.5, 9, and 18 nM / kg showed a statistically significant reduction in triglycerides compared to the diabetic control group. The effect was maintained, with a slight improvement on day 28. Surprisingly, the efficacy of compound 2 in reducing triglyceride levels was found to be similar to that shown by tilzepatide, at approximately 20 times the dose of compound 2.When examining the effects of compound 2 on reducing HbA1c and triglyceride levels, we were surprised to observe an improvement in the effect on day 28 compared to day 14. For example, the reduction in HbA1c on day 29 at doses of 9 nM / kg and 18 nM / kg was more than 40% greater than the reduction on day 14. In comparison, tilzepatide at a dose of 180 nM / kg showed only a slight improvement in HbA1c reduction between day 14 and day 28.

[0193] Example 23: cAMP assay In vitro efficacy measurements were performed using a cAMP assay. Activation of G protein-coupled receptors (GPCRs) following ligand binding initiates a second messenger cascade that leads to a cellular response. Signaling by GLP-1R and GIP-R involves activation of adenylyl cyclase and cAMP production. Cellular cAMP production was determined using the cAMP Hunter® eXpress GPCR assay (Eurofins DiscoveRx).

[0194] In a cell-based cAMP assay, compound 2 had a semi-maximal effective concentration of 4.1 nM in GLP-1R-expressing cells, while tilzepatide with a tilzepatide / compound 2 ratio of 1.68 had a semi-maximal effective concentration of approximately 6.86 nM. In GIPR-expressing cells, the semi-maximal effective concentration of compound 2 was 2.3 nM, while tilzepatide with a tilzepatide / compound 2 ratio of 0.81 had a semi-maximal effective concentration of 1.89 nM. These results indicate that representative compound 2 is a potent inhibitor of both GLP-1 and GIP receptors.

[0195] Example 24: Effects on blood glucose levels, body weight, and food intake. The effect of the compounds of the present invention on blood glucose levels was studied in mice. This study was conducted in a type 2 diabetic mouse (db / db) model. The animals were divided into eight treatment groups (n = 6 per group), namely a diabetic control group, and treatment groups with Compound 2 to Compound 7 (3 nM / kg) and tildepati (10 nM / kg). Compound 1 (6 nM / kg) and Compound 2 (6 nM / kg) were compared with tildepati (59 nM / kg) in separate tests (n = 5 for treatment). Baseline blood glucose levels were measured in all animals. Each of the test compounds was administered subcutaneously to all animals. Blood glucose levels were measured at 4 hours, 12 hours, 24 hours, 48 hours, 72 hours, and 96 hours after treatment. Delta blood glucose values (mM) were calculated. The results are provided in Table 4. Similarly, body weight changes and cumulative food consumption were measured at 96 hours after treatment. The results are provided in Table 5 below.

Table 10

Table 11

[0196] The effects of Compound 2 and Compound 7 on blood glucose levels were further tested in mice at doses of 10 nM / kg and 30 nM / kg, respectively, where blood glucose levels were measured at 4 hours, 8 hours, 12 hours, 24 hours, 48 hours, and 72 hours after treatment and compared with tildepati (90 nM / kg). The results are provided in Table 6 below. Similarly, body weight changes and cumulative food consumption were measured at 72 hours after treatment. The results are provided in Table 7 below.

Table 12

Table 14

[0197] The results demonstrate that the compounds of the present invention can effectively reduce T2D blood glucose levels. The results also indicate that the compounds of the present invention are effective over a long period. Surprisingly, the effect of compound 2 on blood glucose reduction was similar to that shown by tilzepatide at a dose approximately nine times higher than that of compound 2. Furthermore, the effectiveness was maintained for 72 hours. Similarly, the compounds showed a statistically significant reduction in food intake and body weight.

[0198] In another study, the effects of compounds 2, 8, 9, and 10 on blood glucose levels, food intake, and body weight were investigated in mice. This study was conducted using a type 2 diabetic mouse (db / db) model. The animals were divided into five treatment groups (n=6), a diabetic control group, compound 2 (10 nM / kg), compound 8 (10 nM / kg), compound 9 (10 nM / kg), and compound 10 (10 nM / kg). Baseline blood glucose levels were measured in all animals. All animals received subcutaneous administration of the test compound. Blood glucose levels were measured at 4, 12, 24, 48, 72, and 96 hours post-treatment. Delta blood glucose levels (mM) were calculated. The results are shown in Table 8. Body weight change and cumulative food consumption were measured at 96 hours post-treatment. The results are shown in Table 9. Similarly, the effects of compounds 11-15 on blood glucose levels, food intake, and body weight were investigated in separate studies, except for compound 13. The results are shown in Table 8 (effect on blood glucose levels) and Table 9 (effect on body weight and food consumption). [Table 15] [Table 16]

[0199] Compounds 2, 8, 9, 10, 11, and 14 showed a statistically significant reduction in blood glucose levels after treatment. Statistically significant reductions in food intake and body weight were also observed compared to diabetic controls.

[0200] The above results indicate that the compounds of the present invention are potent inhibitors of GLP-1 and GIP receptors and may be effective in treating type 2 diabetes, diabetes with obesity, obesity, and hyperlipidemia.

Claims

1. A polypeptide comprising the following amino acid sequence or a pharmaceutically acceptable salt thereof, Y - During the ceremony X1 is Aib, X2 is Aib, X3 is Lys, and in the formula, the amino (ε-amino) group of the Lys side chain is acylated at the following part: {-U-W-Y-Z In the formula, U is -C(O)-CH ２ -O-(CH ２ ) ２ -O-(CH ２ ) ２ } is -NH-, where} is the attachment point with the base W, W is -C(O)-C(CH3)2-NH-], where ] is the attachment point with group Y, Y is -C(O)-(CH ２ ) ２ -CH(COOH)NH--, and -- is the attachment point to group Z. Z is -C(O)-(CH ２ ), ｎ -COOH or -C(O)-(CH ２ ), ｎ -CH ３ wherein n is an integer of 14 to 20, X4 is Ile, In the formula, the acid group of the C-terminal amino acid is either a free carboxylic acid group or is amidated as a C-terminal primary amide. A polypeptide or a pharmaceutically acceptable salt thereof.

2. The following amino acid sequence: i) Tyr Aib Glu Gly Thr Phe Thr Ser Asp Tyr Ser Ile Aib Leu Asp Lys Ile Ala Gln Lys Ala Phe Val Gln Trp Leu Ile Ala Gly Gly Pro Ser Ser Gly Ala Pro Pro Pro Ser-NH ２ (SEQ ID NO: 5) A polypeptide according to claim 1, comprising the same, wherein a pharmaceutically acceptable salt thereof.

3. Z is -C(O)-(CH ２ ) ｎ A polypeptide according to claim 1 or 2, wherein n is -COOH and n is 16 or 18, or a pharmaceutically acceptable salt thereof.

4. W is -C(O)-C(CH ３ ) ２ -NH-] and Z is -C(O)-(CH ２ ) ｎ A polypeptide according to any one of claims 1 to 3, wherein n is -COOH and n is 18, or a pharmaceutically acceptable salt thereof.

5. -U-W-Y-Z are the following groups: 【Chemistry 1】 A polypeptide according to any one of claims 1 to 3, selected from the above, or a pharmaceutically acceptable salt thereof.

6. The polypeptide according to any one of claims 1 to 5, wherein the C-terminal amino acid is amidated as a C-terminal primary amide, or a pharmaceutically acceptable salt thereof.

7. The polypeptide according to any one of claims 1 to 5, wherein the acid group of the C-terminal amino acid is a free carboxylic acid, or a pharmaceutically acceptable salt thereof.

8. A polypeptide selected from the following compounds 1 and 2, or a pharmaceutically acceptable salt thereof: 【Chemistry 2】 In the formula, part A has the following structure 【Transformation 3】 Part B has the following structure 【Chemistry 4】 A polypeptide or a pharmaceutically acceptable salt thereof.

9. A pharmaceutical composition comprising a polypeptide according to any one of claims 1 to 8 or a pharmaceutically acceptable salt thereof, and one or more of a carrier, a diluent, or a pharmaceutically acceptable excipient.

10. A polypeptide according to any one of claims 1 to 8 or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to claim 9, for use as a pharmaceutical agent.

11. A polypeptide according to any one of claims 1 to 8 or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to claim 9, for use in the treatment or prevention of a patient's disease.

12. A polypeptide for use according to claim 11 or a pharmaceutically acceptable salt thereof or a pharmaceutical composition, wherein the disease is selected from the group consisting of hyperglycemia, type 2 diabetes, impaired glucose tolerance, type 1 diabetes, obesity, hypertension, hyperlipidemia, X syndrome, dyslipidemia, cognitive impairment, atherosclerosis, myocardial infarction, coronary heart disease, stroke, inflammatory bowel syndrome, dyspepsia, alcoholism, and gastric ulcer.

13. The polypeptide or a pharmaceutically acceptable salt thereof or pharmaceutical composition for use according to claims 10 to 12, wherein the polypeptide or a pharmaceutically acceptable salt thereof or the pharmaceutical composition is provided simultaneously, separately, or sequentially in combination with one or more additional therapeutic agents in an effective amount.

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